American Journal of Traditional Chinese Veterinary Medicine

AMERICAN JOURNAL OF
TRADITIONAL CHINESE VETERINARY MEDICINE
VOLUME 12, NUMBER 1 February 2017
The Official Journal of the American Association of

Traditional Chinese Veterinary Medicine, the American
Academy of Veterinary Acupuncture, the International
Veterinary Acupuncture Society and the World
Association of Traditional Chinese Veterinary Medicine
copyright © 2017 by AJTCVM All Rights Reserved
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2 AJTCVM Vol 10, No. 2, August 2015
AJTCVM Vol. 12, No. 1, February 2017 3
AJTCVM Ronald Koh DVM

Jianqin Xu DVM
An international peer reviewed scientific Tiffany Rimar DVM
Phil Rogers MVB, MRCVS College of Veterinary Medicine, China
journal of veterinary acupuncture, Chinese
Allen Schoen DVM Agricultural University, China
herbal medicine, Tui-na and Food Therapy
published by the AATCVM Cheryl Schwartz DVM Jianliang Zhang PhD
Justin Shmalberg DVM Roswell Park Cancer Institute
ISSN 1945-7677 (print) acid free Susan Wynn DVM Buffalo, NY
ISSN 1945-7693 (electronic) Madeline Yamate DVM
Jose Zilberschtein DVM, MS, PhD
The Official Journal of the American Board of Scientific Reviewers College of Veterinary Medicine
Association of Traditional Chinese Keum Hwa Choi DVM, PhD, OMD Murcia University, Spain
Veterinary Medicine (AATCVM), College of Veterinary Medicine,
the American Academy of Veterinary University of Minnesota AATCVM
Acupuncture (AAVA), the International American Association of
Veterinary Acupuncture Society (IVAS) Roger Clemmons DVM, PhD
Traditional Chinese Veterinary
and the World Association of Traditional College of Veterinary Medicine,
University of Florida Medicine
Chinese Veterinary Medicine (WATCVM).
Website: http://www.aatcvm.org
Mission Statement Christine Egger DVM, MS, DACVA Mission Statement
The mission of AJTCVM is to provide College of Veterinary Medicine, Support TCVM practitioners world-wide
practitioners, students and researchers with University of Tennessee by providing an on-line Forum for daily
the highest level of peer-reviewed articles case discussions, a library of TCVM
Maria Grant MD
on veterinary acupuncture, Chinese herbal references and materials for presentations;
College of Medicine
medicine, Tui-na and Food Therapy to Promote TCVM research by funding small
Indiana University
promote the generation and publication of grants and providing assistance with study

research on the efficacy of these therapies Jianguo Gu MD, PhD design and manuscript preparation; Publish
in animals and to support the practice of College of Medicine, a peer-reviewed scientific TCVM journal;
evidence-based medicine. University of Cincinnati Develop scientific guidelines for TCVM
diagnosis, treatment and prevention of
AJTCVM Staff Shining Guo, DVM PhD disease.
Editor-in-Chief College of Veterinary Medicine AATCVM, PO Box 141324
Judith Saik DVM, DACVP South China Agricultural University Gainesville, FL 32614

Jorge Herndandez DVM, PhD
Executive Editors
College of Veterinary Medicine AAVA
Aituan Ma MS, PhD
University of Florida American Academy of Veterinary
Carolina Medina DVM, DACVSMR
Neal Sivula DVM. PhD Acupuncture
Songhua Hu DVM, PhD Website: http://www.aava.org
Huisheng Xie DVM, MS, PhD College of Veterinary Medicine Zhejiang
Mission Statement
Associate Editors University, China To improve animal health care by the
Signe Beebe DVM advancement of veterinary acupuncture,
Min Su Kim DVM, PhD
G. Reed Holyoak DVM, PhD, DACT College of Veterinary Medicine, Chonbuk traditional Chinese veterinary medicine
Lindsey Wedemeyer MA, VetMB, National University, Korea and traditional Asian medicine through
MRCVS education, research and leadership.
Jian Kong MD, MS, MPH AAVA, PO Box 1058
Assistant Editors Harvard Medical School Glastonbury, CT 06033
Bruce Ferguson DVM, MS Harvard University
Erin Mayo DVM IVAS
Carla Pasteur DVM Albert Leung MD International Veterinary
Karen Spracklen DVM School of Medicine Acupuncture Society
Laura Smith DVM UC San Diego Website: http://www.ivas.org
Gregory Todd DVM
Daiqing Liao PhD Mission Statement
Marisa Chan Jing Ying BSc, BVMS
College of Medicine, The Mission of the International

Consulting Editor University of Florida Veterinary Acupuncture Society is to
Cheryl L Chrisman DVM, MS, EDS, provide, promote and support veterinary
Hanru Liu DVM, MS acupuncture and related treatment
DACVIM
College of Veterinary Medicine, modalities through quality basic, advanced

Graphic Design Southwest China University, China and continuing education; internationally
Autumn Fruchtman, BA recognized certification for veterinarians;
Zhigang Lu MD, PhD
and support responsible research.
Advertising Memorial Sloan Kettering Cancer Center,
New York, NY IVAS 1730 S. College Ave, Ste 301
Suzy Brannan DVM
Fort Collins, CO 80525
polargators@gmail.com
K. Gary Magdesian DVM,

Webmasters ACVIM, ACVECC, ACVCP WATCVM
AATCVM: ts@ajtcvm.org College of Veterinary Medicine, World Association of Traditional
AAVA: webmaster@aava.org University of California, Davis Chinese Veterinary Medicine
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IT Support College of Veterinary Medicine,
Zhen Zhao MS Vision Statement:
Washington State University
Advisory Board The WATCVM supports the education,
Constance DiNatale DVM Weerepongse Tangjitjaroen DVM, PhD research, and practice of Traditional
Joyce Harman DVM College of Veterinary Medicine Chinese Veterinary Medicine worldwide.
Dan Hawkins DVM, MS, DACVS Chiang Mai University, Thailand
AJTCVM Vol. 12, No. 1, February 2017 i

Animal Use and Care: All research studies involving animals must have been performed in compliance with guidelines outlined in the
Animal Welfare Act, US Public Health Service Policy on the Humane Care and Use of Laboratory Animals, NRC Guide for the Care and
Use of Laboratory Animals, or Guide for the Care and Use of Agricultural Animals in Agricultural Research and Teaching (Fass 1999) or
with equivalent guidelines. A manuscript containing information that suggests that animals were subjected to adverse, stressful, or harsh
conditions or treatments will not be considered for publication. Manuscripts describing studies that involved the use of animals, including
studies that involved the use of privately owned animals (e.g. animals owned by clients, staff members, students, or private entities) must
include a statement that the study protocol was reviewed and approved by an appropriate oversight entity (e.g. an animal care and use
committee or institutional review board) or was performed in compliance with institutional guidelines for research on animals. Manuscripts
describing prospective studies that involved privately owned animals must include a statement indicating that owner consent was obtained.
Conflict of Interest: Authors are expected to acknowledge all sources of funding or financial support and to disclose to the Editor-in-Chief
any financial interests (including ownership, employment, consultancy arrangements, and service as an officer or board member) they have
with companies that manufacture products that are the subject of their research or with companies that manufacture competing products.
Copyright: All material printed in AJTCVM is covered by copyright. All rights reserved. No portion may be republished, reproduced,
photocopied, displayed or transmitted without prior written permission from the publisher. Address permission requests to: Dr. Judith Saik
at: saikj@ajtcvm.org
AJTCVM Statement of Informed Consent: Animal owners (clients) and their animals have a right to privacy that should not be infringed
upon without informed consent. Identifying information, including animals’ names, initials or medical record numbers, should not be
published in written descriptions or photographs unless the information is essential for scientific purposes and the animal owner gives
written informed consent for publication. Clients must consent to all photographs of animals included in a manuscript. Pedigrees should not
be published without signed consent from the involved breeders. Identifying details should be omitted if they are not essential. If identifying
characteristics are altered to protect anonymity, such as in genetic pedigrees, authors should provide assurance that alterations do not distort
scientific meaning and editors should so note the changes. Authors should identify individuals who provide writing assistance and disclose
the funding source for this assistance.

About the Cover

In 2017 the covers of AJTCVM will boast the color gold to honor the Earth Element.
Longji Rice Terraces
This issue’s cover honors and celebrates the Earth
Element with an image of the Longji Rice Terraces.
Rice can support the Earth Element by nourishing
the Spleen. This rice paddy field is located just
north of Guilin in the Guangxi Province and covers
about 66 square kilometers. Construction of the
fields began during the Yuan Dynasty (1271–1368)
and completed in the Qing Dynasty (1644–1911).
Ancient Zhuang and Yao villages still exist in the
areas surrounding the rice terraces. The name Longji,
meaning Dragon’s Backbone, received its name due
to the resemblance of the terraced fields to a dragon’s
back. The Longji Rice Terraces have become a
popular tourist destination attracting both hikers and
photographers.
Errata Corrige
The Editors apologize for the incorrect statement in the “TCVM Around the World” column stating that the International
Veterinary Acupuncture Society (IVAS) provides chiropractic certification. IVAS provides veterinary acupuncture
certification; not chiropractic certification. AJTCVM 11(2) 2016:3-5.
ii AJTCVM Vol. 12, No. 1, February 2017

The American Journal of Traditional Chinese

Veterinary Medicine (AJTCVM)
An Internationally Peer-Reviewed Journal of Veterinary Acupuncture,
Chinese Herbal Medicine, Tui-na and Food Therapy
February 2017 Vol. 12 No. 1

Table of Contents
Editorial
Black Box Theory and Diagnostic System of Traditional Chinese Veterinary Medicine................................................1
Huisheng Xie DVM, MS, PhD
TCVM News and Educational Opportunities.................................................................................................................................5

AAVA Updates, Letter from the President.....................................................................................................................................7
AATCVM Updates, Letter from the President.............................................................................................................................9
IVAS Updates, Letter from the President......................................................................................................................................13
WATCVM Updates, Letter from the President.............................................................................................................................17
Basic Science Studies

A Randomized, Blinded, Double Controlled Study of the Effects of Modified Ma Xing Shi Gan Tang on Mortality,
Recovery and Serum Superoxide Dismutase, Glutathione Peroxidase and Malondialdehyde in Chickens with
Infectious Bronchitis.....................................................................................................................................................................19
Yongping Yan DVM, PhD, Xiaodan Wang DVM, PhD, Ruiwen Li DVM, PhD, Xiu-hui Zhong DVM, PhD
Safety Evaluation of Modified Si Jun Zi Tang, Assessment of Controlled Randomized, Oral Acute and Subchronic
Mouse Toxicity Studies...................................................................................................................................................29
Shuang Ma DVM, MS, Aituan Ma DVM, MS, PhD
A Randomized and Controlled Experimental Study on the Effects of Epimedium-Propolis Adjuvant

on Immune Function in Mice, Rabbits and Chickens...................................................................................................39
Yuanliang Hu DVM, PhD, Jiaguo Liu DVM, PhD, Dalu Song DVM, Zhigang Xu DVM
Clinical Studies
A Randomized, Controlled Study Comparing Da Xiang Lian Wan to Metronidazole in the Treatment of Stress Colitis in
Sheltered/Rescued Dogs..................................................................................................................................................45
Margaret P Fowler DVM, CTCVMP, MS, Deng-Shan Shiau PhD, Huisheng Xie DVM, PhD
Case Series
The Therapeutic Effect of Acupuncture and Chinese Herbal Medicine in 12 Dogs with Hyperadrenocorticism...........55
Ronald Koh DVM, MS, Huisheng Xie DVM, PhD, May-li Cuypers DVM, DACVIM
AJTCVM Vol. 12, No. 1, February 2017 iii

Review
Review of Evidence-Based Clinical and Experimental Research on the Use of Acupuncture and Chinese Herbal Medicine
for the Treatment or Adjunct Treatment of Cancer............................................................................................................69
Huisheng Xie DVM, MS, PhD, Betsy Hershey DVM, DACVIM (Oncology), Aituan Ma DVM, MS, PhD
Brief Communications
Preliminary Evaluation of the Effect of Acupuncture on Acoustic Myographic Recordings in Five Sport
Horses...................................................................................................................................................................79
Esther M. Millares DVM, Helga Bollingberg-Soerensen Sarah S. Le Jeune DVM, DACVS, DACVSMR
Pearls from TCVM Practice

Feline Inappropriate Elimination and Lower Urinary Tract Inflammation: Theoretical Background and Simple
Management with Acupuncture and Chinese Herbal Medicine......................................................................................85
Bruce Ferguson DVM, MS
Laser Acupuncture and Local Laser Therapy in Veterinary Medicine with Overview of Applied Laser Types and
Clinical Uses..................................................................................................................................................................................89
Uwe Petermann DVM
Selected Abstracts
Acupuncture and endorphins.........................................................................................................................................................8
Effects of synchronous or asynchronous electroacupuncture stimulation with low versus high frequency on spinal
opioid release and tail flick nociception......................................................................................................................................102
Herbal Materia Medica

Gui Zhi (Cinnamomum)....................................................................................................................................................103
Caili Zhang DVM, MS
Chinese Herbal Medicine Spot Light

Tao Hong Si Wu Tang (Four-Substance Decoction with Safflower and Peach Pit)....................................................104
Signe E Beebe DVM
Classified Advertisements
IVAS..................................................................................................................................................... inside front cover
AAVA.....................................................................................................................................................inside front cover
Mayway...................................................................................................................................................................6
Golden Needle..................................................................................................................................................................6
Acuro...................................................................................................................................................................11
Chi Institute Ophthalmology Course..............................................................................................................................12
Eastern Currents.............................................................................................................................................................12
Golden Flower................................................................................................................................................................18
Margaret Fowler Classified Ad: Practice for Sale.........................................................................................................54
Alt-Vet by Mitsie Vargas..............................................................................................................................................68
Kan Herb........................................................................................................................................................................38
Pet Tao............................................................................................................................................................................77
iv AJTCVM Vol. 12, No. 1, February 2017

Chi Institute International TCVM Conference................................................................................................................78

IVMI Veterinary Medical Manipulation Program.........................................................................................................84
PCLAC Book................................................................................................................................................................102
AJTCVM Classified Advertising Information and Rates............................................................................................103
WATCVM...............................................................................................................................................inside back cover
AATCVM...............................................................................................................................................inside back cover
Instructions to Authors – visit www.ajtcvm.org (authors tab)
AJTCVM Vol. 12, No. 1, February 2017 v

Editorial
Black Box Theory and Diagnostic System of

Traditional Chinese Veterinary Medicine
Huisheng Xie DVM, MS, PhD
History of Black Box functions internally by the relationship between the

The initial idea of Black Box can be traced to exterior appearance of something to its characteristics and
two-port networks (quadripoles) that were created by behavior within (interior/inner).
a German mathematician, Franz Breisig, in 1921.1 For example, the school of behaviorism sees the
In 1941, twenty years later, Wilhelm Cauer, another human mind/brain as a Black Box in psychology.5 Since
German mathematician and scientist, applied this idea in the brain cannot be opened to be examined, a psychologist
electronic circuit theory.2 Norbert Wiener, an American can design a series of questions for a patient (input),
mathematician and philosopher, defined a Black Box to and the responses from a patient (output) are analyzed
be an unknown system that was to be identified using to decipher the type of behavior issues present. Another
the techniques of system identification in 1948.3 Since example is that the earth can be treated as a Black Box
then, the Black Box theory has been widely applied to when undertaking oil exploration. After exploration tests
philosophy and science. are conducted on the surface of the Earth (input), the
signals from the surface (output) are analyzed to determine
What is the Black Box? potential oil field locations.
A Black Box is a system or object which is purely
external or phenomenological and its internal constitution White Box and Medicine
and structure is unknown or irrelevant to being opened With a White Box, we can see what is inside the
for inspection.4 The opposite of a Black Box is a White box (Figure 1). All modern diagnostic devices including
Box, that is a system where the internal constitution and MRI, CT scan, radiographs and ultrasound help clinicians
structure can be viewed. A White Box is also referred to as visualize the interior of the body and give an accurate
a clear box, transparent box, or glass box. diagnosis (bony changes, fracture, neoplasia and other
As illustrated in Figure 1, a White Box is transparent, pathologic changes); thus, they are the White Box
like glass, as we can see all that is inside the box. On the approach. Another example is exploratory laparotomy. The
contrary, we cannot see what is inside the Black Box. abdominal exploratory surgery is often used to investigate
How the Black Box functions internally, however, can an inconclusive mass or other gastrointestinal disease
be tested and defined in terms of its “stimuli inputs” and in veterinary medicine. After the abdomen is surgically
“output reactions” (Figure 2).4 In other words, an observer opened, a veterinarian is able to view pathological changes
or investigator is able to figure out how the Black Box (such as masses), and a biopsy sample is collected when
Figure 1: White vs Black Boxes Figure 2: How the Black Box works
necessary for further diagnosis. of Heat or Fire tend to ascend, thus their pathological
damage is not only in the Liver, but also in the eyes that
The Traditional Chinese Veterinary Medicine (TCVM) are the Exterior portion of the Liver System. TCVM
Diagnostic System practitioners, therefore, can predict that the condition is
When TCVM diagnosis was developed several Liver Heat or Fire (interior) when they see redness and
thousand years ago, ancient healers did not have modern swellings of the eyes (exterior).
diagnostic tools such as laboratory tests, radiography, Other examples of exterior observation revealing
ultrasound, scintigraphy and magnetic resonance imaging internal TCVM Patterns include gingival bleeding/
(MRI). Instead, the ancient medical practitioners could only ulceration with Stomach Fire, tongue ulcers with Heart
rely on their senses to gather information about a patient’s Fire, nasal discharge with Lung disorders, diarrhea
condition. The practitioners visually evaluated a patient’s with Spleen Qi Deficiency and congenital disease with
behavior and body appearance to identify abnormalities. Jing Deficiency. In the first 2 exterior observations, the
They used their hands to feel the pulse and palpate the redness and swelling associated with gingival bleeding
body. Their senses of hearing and smell allowed them to or ulceration and tongue ulcers corresponds to Heat or
identify abnormal noises or odors coming from the patient. Fire. Since the Stomach Channel runs through the gingiva
Over generations, TCVM practitioners established Si-zhen and the Heart opens into the tongue, the interior TCVM
(“four diagnostic methods”) which provided a systematic Patterns associated with these external clinical signs is
method for collecting clinical data (external information Stomach Fire and Heart Fire, respectively. The nose is
from the patient) in order to make the correct diagnosis the opening for the Lung, therefore, nasal discharge/
(Table 1).6 The theoretical principle behind Si-zhen is that congestion (exterior) is associated with Lung disorders
changes occurring within the Interior of the body will be (interior). A watery, loose stool (exterior) is a diagnostic
evident on the Exterior due to the connections among sign for Spleen Qi Deficiency (interior) as the primary
the organs. Thus, a practitioner can examine the Exterior function (Qi) of the Spleen is to digest/absorb food and
signs to understand the condition of the body’s Interior move fluid. Deficient Spleen Qi fails to digest/absorb food
Zang-Fu organs. This is well documented as “To know the and move fluid, leading to excessive dampness and watery
inside by examining the outside” in the ancient traditional diarrhea. Finally, a young white kitten born deaf, suggests
Chinese medical classical text Huang Di Nei Jing (Yellow a Kidney problem (hearing functions are associated with
Emperor’s Classic of Medicine).7 For example, redness the Kidney), specifically a Kidney Jing Deficiency.
and swelling of the eyes may indicate a Liver Heat Pattern
because eyes are associated with the Liver and redness and TCVM Etiology
swelling correspond with Heat or Fire. The characteristics According to TCVM, the body normally exists in a
Table 1: The Four Diagnostic Methods (Si-Zhen)
Western Medicine
Si-zhen TCVM Examination
Counterpart
• Examine the tongue characteristics and color patterns

• Observe the Shen (the Mind-state)
Inspection Wang Observation
• Observe the general body condition
• Evaluate the quality and luster of the hair and skin
• Listen to the voice quality (weak, muffled, strong and/or robust)

Hearing • Listen to breathing Auscultation
Wen
Smelling • Auscultate heart and lungs Olfaction
• Note odors from the body, breath, ears
• Question owner about medical history
Inquiring Wen • Question owner about temperature-preferences, diet, personality, activity, Interrogation
clinical signs
• Feel the pulses
• Palpate the Meridians
Palpation Qie • Palpate the back-Shu (association) points and front-Mu (Alarm) points for Palpation
sensitivity
• Palpate abdomen, lymph nodes, joints
2 AJTCVM Vol. 12, No. 1, February 2017

state of equilibrium between the external environment and principles can be advantageous for disease diagnosis.
the Zang-Fu organs. To maintain this equilibrium, the body Conventional Western diagnostic tests and physical
regularly readjusts itself to maintain normal functionality exam complement the traditional Chinese history and
as the various external and internal forces fluctuate. physical exam. A Western diagnosis can become part of
Disease results when certain forces overwhelm the body’s the TCVM diagnosis, and a TCVM diagnosis can further
capacity to adjust and disrupt the delicate balance. The clarify a Western diagnosis. For example, radiographs are
factors which impair the body’s equilibrium and cause an invaluable tool to visualize the joints and may provide
disease are called Pathogens or Xie Qi. The Xie Qi exists a Western diagnosis of degenerative joint disease or
external to the body and includes allergens, seasonal or arthritis. Chinese medicine, however, can further define
environmental changes and infectious organisms such as which type of arthritis based upon the patient’s collective
viruses, bacteria, and fungi. These etiologic factors invade clinical signs.
the body through the skin, mouth, or nose and cause
imbalance which results in disease. The TCVM Diagnostic System and Black Box
Different pathogens produce different signs within the “To know the inside by examining the outside” is the
body, so a TCVM practitioner can diagnose the etiology by fundamental principle of Traditional Chinese Veterinary
analyzing the clinical manifestations of a disease. This is Medicine (TCVM) diagnostics. Thus, the animal body is
known as “seeking the cause of a disease by differentiating treated as a Black Box. A TCVM practitioner relies on a
clinical signs” or Shen Zheng Qiu Yin.8 In addition, the patient’s external information (clinical signs, tongue and
treatment principle depends upon the cause of the disease. pulse) to make a diagnosis and predict how the internal
This is otherwise known as “treatment according to the organs function or malfunction. This diagnostic system
cause” or Shen Yin Shi Zhi.9 It is the TCVM practitioner’s is also called Bian Zheng involving identifying pertinent
role to recognize the exterior clinical signs resulting from clinical signs (Bian) and determining the Pattern of illness
a certain Pathogen and then administer a specific treatment (Zheng).
to resist the previously diagnosed Pathogen. For example, TCVM is the Black Box approach. Initially, a Black
the TCVM practitioner would recognize that a sick Box was used as a computing device that returned an
horse exhibiting alternate limb lameness is affected by output after giving an input (Figure 2). In TCVM, all of the
pathogenic Wind and Damp and would apply a treatment pathogenic factors (allergen, virus, bacteria and fungus)
that eliminates Wind and Damp. are the input (stimulus). Zheng Qi (the collective anti-
TCVM diagnosis relies heavily on gathering pathogenic power) of the animal body (Black Box) will
information about clinical signs and then determining how respond to the invasion of Xie Qi. All of the output data
they fit with known Pathogens, therefore, combining both (tongue, pulse and clinical signs) from a patient can be
conventional and traditional Chinese medical diagnostic analyzed and a final TCVM Pattern can be differentiated
Fever
Figure 3: The Black Box approach of the TCVM Diagnosis

and determined (Figure 3). Thus, for an effective Chinese medical treatment, the
TCVM practitioner must accurately identify the signs
Clinical Application of the TCVM as a Black Box indicating the kind of imbalance and not rely solely on the
Identifying relevant clinical signs is a common task in Western diagnosis.
Western medical diagnosis; however, the concept of Zheng
(Pattern of disease) is a much more formalized system in Conclusion
TCVM. In TCVM, the diagnosis is the TCVM disease The TCVM approach is like the Black Box. External
Pattern (Zheng). A particular TCVM diagnosis is assigned information (clinical signs) displayed by patients is used
to specific combinations of clinical signs resulting from to diagnose the correct TCVM Pattern which enables the
particular imbalances within the patient. veterinarian to select an appropriate treatment correctly
A Western medical disease diagnosis analyzes matched to the pattern. It is a very useful approach,
significant clinical signs, laboratory tests and diagnostic especially in clinical cases when a Western diagnosis is
images to find the underlying cause. Although Western not available or is unable to pinpoint the etiology of a
practitioners do acknowledge that some diseases can disease.
produce stereotypical physical manifestations, such as
Cushing’s disease in dogs; these disease patterns are References
less specific than in TCVM. The same Western medical 1. Belevitch, V, “Summary of the history of circuit
diagnosis may correspond to different TCVM Patterns. theory”, Proceedings of the IRE: May 1962: 50 (5):
Consider the following two examples: 848-855.
2. W. Cauer. Theorie der linearen Wechselstrom-
Case Example One: A Western medical practitioner schaltungen, Vol.I. Akad. Verlags-Gesellschaft
diagnosed two horses with bloody diarrhea as having Becker und Erler, Leipzig, 1941.
Salmonella colitis because fecal cultures were positive 3. Wiener, Norbert. Cybernetics: or the Control and
for Salmonella species. The first horse was a two-year- Communication in the Animal and the Machine. page
old Thoroughbred filly who presented with profuse xi, MIT Press. 1948.
hemorrhagic diarrhea which had begun three days prior 4. Mario Bunge. A general Black-Box theory.
to presentation. She had a red tongue and rapid pulse. The Philosophy of Science 1963: 30 (40): 346-358.
second horse was an eighteen-year-old Thoroughbred 5. Jay Friedenberg, Gordon Silverman. Mind as a
mare who presented with a twenty-one day history of Black Box: The Behaviorist Approach. in Cognitive
bloody diarrhea. The mare had a pale tongue and weak Science: An Introduction to the Study of Mind. Sage
pulse. Although both horses were diagnosed as having Publications 2006: 85-88.
Salmonella colitis, their TCVM diagnoses differed; the 6. Xie H, Preast V. Traditional Chinese Veterinary
first horse had an Excess Heat Pattern because of the red Medicine-Fundamental Principles (2nd edition).
tongue, rapid pulse, young age and acute onset, while the Reddick, Florida: Chi Institute, 2013: 205-208.
second horse had Spleen Qi Deficiency based upon her 7. Chapter 47 of Ling Shu (Miraculous Pivot), Huang
pale tongue, weak pulse, old age and chronic diarrhea. Di Nei Jing (Yellow Emperor’s Classic of Medicine),
(In Chinese, originally published in 475-221 BC).
Case Example Two: A Western medical practitioner 8. He Y, Liu W. New Edited Traditional Chinese
used allergy testing to diagnose two dogs with atopic Medicine-Fundamental Principles (Xin Bian Zhong
dermatitis. One dog was a friendly and high energy Yi Ji Chu Li Lun) (In Chinese). Beijing, China:
two-year-old castrated male Labrador retriever with a Beijing Medical University & China Xie He Medical
Fire personality. He presented with constant pruritus University Press 1996:128-129.
and showed Heat signs (red tongue, a bounding pulse 9. Liu Z, Xu J ed. Traditional Chinese Veterinary
and cool-seeking behaviors) without Deficiency signs Medicine (Zhong Shou Yi Xue), 3rd Ed (In Chinese).
(weakness). The second atopic dog was a thirteen-year- Beijing, China: China Agricultural Press 2005:48-51.
old spayed female bichon frise that has chronic allergies
since she was young. She seemed less itchy as a geriatric
dog, but was weaker overall and still developed pyoderma
and pustules on her ventrum. Her tongue was pale and dry.
Her owner reported that she preferred comfortable places,
drank a lot of water, and her body was slightly warm to the
touch. From a Zheng perspective, these two dogs have two
different Patterns of diseases, even though they both have
the same Western diagnosis. The young Labrador showed
signs of Excess due to an invasion of Wind-Heat, whereas
the older dog had a Kidney Yin/Liver Blood Deficiency.

TCVM Around the World and Educational

Opportunities
TCVM Courses, Conferences and Acupuncture” with Dr. Uwe Petermann in Fort Collins,
Workshops in Acupuncture, Chinese CO at the Museum of Discovery, May 5-7, 2017. For more
Herbal Medicine, Tui-na and Food information visit www.ivas.org or contact the IVAS office
Therapy at office@ivas.org.
The Chi Institute of the United States provides small The Qi Academy of Germany offers acupuncture, herbal
animal, equine or mixed animal acupuncture practice. For medicine, food therapy and Tui-na courses. For more
more information, please visit: http://www.tcvm.com/ information, please visit: http://qiacademy.net/.
Programs.aspx.
The Chi Institute of the United States provides basic
The Chi Institute and the WATCVM will co-host the and advanced programs on acupuncture, Chinese
19th Annual international TCVM Conference in Toronto, veterinary herbal medicine, food therapy and Tui-na. For
Canada, August 18-20, 2017. Visit www.tcvm.com for more information, please visit: http://www.tcvm.com/
more information. Programs.aspx.
The Chi Institute of Europe offers a certified veterinary The AAVA will hold its annual meeting March 24-26,
acupuncture program in both English and Spanish Classes 2017 in Savannah, GA. The FAAVA exam will take place
at Madrid, Spain. For more information visit: www.mvtc. March 23, 2017 in Savannah, GA as well. Visit www.aava.
es. org for more information.
The Chi Institute of Australia offers the certified Pulse Controlled Laser Acupuncture Concept
veterinary acupuncture program. For more information (PCLAC) provides advanced course on veterinary laser
visit: www.tcvm.com.au. acupuncture, presented by Dr. Uwe Petermann. For more
information, please visit: http://www.akupunkturtierarzt.
The College of Integrative Veterinary Therapies de/Akupunkturtierarzt/heart_disease.html.
(CIVT) provides veterinary Chinese herbal medicine and
acupuncture courses. For more information, please visit The Academic of Veterinary Homeopathy provides
http://www.civtedu.org/. certified veterinary homeopath course. For more
information, please visit: http://theavh.org/certification/.
The International Veterinary Acupuncture Society
(IVAS) will present “Introduction to Chinese Food The Veterinary Botanical Medicine Association
Therapy” with Dr. Connie DiNatale and Dr. Linda Boggie (VBMA) will offer herbalist certification. For more
at the Museum of Discovery in Fort Collins, CO. February information, please visit: http://www.vbma.org/
11-12, 2017. For more information visit www.ivas.org or continuing-education.html.
contact IVAS at office@ivas.org.
The Canine Rehabilitation Institute provides
The International Veterinary Acupuncture Society canine rehabilitation course, please visit http://www.
(IVAS) and the Australian Veterinary Acupuncture Group caninerehabinstitute.com/Calendar-list.html for greater
(AVAG) will present a joint Congress on Veterinary details on the course.
Acupuncture, August 23-26, 2017 at the Hilton Cairns in
Cairns, Australia. Join us in the Land Down Under. For The American Veterinary Chiropractic Association
schedule, registration and more information visit www. (AVCA) offers a basic program of veterinary chiropractic
ivas.org certification courses. For more information, please visit:
http://www.veterinarychiropractic.ca/basic-program.
The International Veterinary Acupuncture
Society (IVAS) will present “Auricular Medicine/Ear The Healing Oasis Wellness Center offers courses on

spinal manipulative therapy, veterinary massage and

rehabilitation therapy. For more information, please visit:
http://healingoasis.edu/post-graduate-programs/.
If you have any TCVM News Around the World that you
would like to share, please contact Dr. Caili Zhang at:
Caili@tcvm.com.
Biography of Dr. Caili

Zhang, TCVM Around
the World Moderator
Dr. Caili Zhang DVM,
MS, attended the College
of Traditional Chinese
Veterinary Medicine at
the Agriculture University
of Hebei and received
her Bachelor’s degree of
Veterinary Medicine in 2011
and her DVM in 2012. She went on to earn a Master’s
degree at the College of Veterinary Medicine in 2014.
During her 3 years of graduate study, she mainly focused
on the effects and associated mechanism of Lycium
Barbarum Polysaccharides on hamster spermatogenic
injury induced by Diethylstilbestrol (DES). She has
published 6 papers in TCVM academic journals and was
a presenter at the TCVM Annual Conference hosted by
the Chinese Association of Animal Science and Veterinary
Medicine at Haerbin. She works as a member of the Chi
Institute.
Chinese Herbs & Herbal products since 1969

AAVA Updates
Keum Hwa Choi DVM, PhD, CVA, OMD, LAc (MN), Diplomate NCCAOM (Oriental Medicine)
President, AAVA
I am Keum-Hwa Choi. I have been elected as clinician.

the 2016-2017 President of the American Academy of The last 40 years of Traditional Eastern Veterinary
Veterinary Acupuncture (AAVA). I express great thanks Medicine (TEVM) has been practiced and slowly
for Dr. Patricia Baily, our outgoing president who recognized in the USA becasue of many devoted pioneers,
accomplished many important agendas during her term. clinicians, scholars, institutes and publics. During this time,
I am also thankful for present and past board of director AAVA has been a core player promoting the discipline of
(BOD) members who devote their time and effort to this TEM. However, non-conventional medicine is still being
community. challenged by the mainstream medical society due to the
I was once very skeptical of Traditional Eastern lack of scientific evidence, skillful doctors, and support
Medicine, especially acupuncture. I vividly remember the from educational systems. As a small step for overcoming
day I had acupuncture for the first time. One day, I hurt this challenge, I set three major missions during my term
my knee after hitting it against a cement wall. I went to the which are as follows;
Western Medical doctor, but the pain and swelling were not 1. To carry on the AAVA’s current missions to accomplish
alleviated. One of my colleagues suggested acupuncture. them.
While driving home after speaking with this colleague, I 2. To improve the educational system for TEM and seek
saw a sign for a “Traditional Korean Acupuncture Clinic”. to provide quality education to veterinarians through
I went into the clinic and waited, expecting an older conferences, CE courses, webinars and other mediums.
doctor with a long grey beard. To my surprise a young, 3. In March of 2016, the American Board of Veterinary
handsome doctor came in and examined my swollen right Specialties (ABVS) did not approve acupuncture as a
knee. During my treatment he put acupuncture needles veterinary specialty. To get a stronger support from the
into my left ankle, this was my uninjuried leg. I felt like he mainstream medical community as well as institutes, I
amputated the wrong leg, I was furious. The next morning will work with our BOD members and our community
I was walking normally again. The swelling and pain were leaders to establish a College of Veterinary Integrative
greatly reduced. It was a turning point in my life. Medicine.
I had the privilege to train with many masters and I encourage you to accept our mission to continue to
institutes of Traditional Eastern Medicine (TEM) in make a great vision possible for the next generation.
Korea, China and the USA. During my journey, the best Last but not least, I would like to thank all of AAVA
event was to have the chance to be trained from several members, BOD, administrators, and supporters. In
masters on Sa-Ahm Traditional Korean medicine which addition, I’d like to let you know that we always welcome
became my most preferred medical mode among several your comments, discussion, advice, and collaboration.
other TEM modes. One thing I stress to practitioners and
students is that the selection of the best medical option Best to all,
for the patient is the most important responsibility as a Dr. Choi

Upcoming AAVA Events
FAAVA Exam: Want to Take the Next Step in your

Professional Acupuncture Development?
The AAVA offers an Advanced Certification
examination! Candidates that successfully pass the
examination are designated as Fellows of the American
Academy of Veterinary Acupuncture (FAAVA). To date,
there are 24 FAAVA’s. The requirements for qualification
American Academy of Veterinary Acupuncture Annual are listed on the AAVA website. The program is based on
Meeting the American Board of Veterinary Practitioner’s Board
The American Academy of Veterinary Acupuncture Certification process and consists of requirements that
will hold its annual meeting in Savannah, Georgia, include clinical experience in acupuncture, continuing
March 24-26, 2017 at The Hyatt Regency Hotel, 2 West education in acupuncture, submission of case reports, and
Bay Street, Savannah, Georgia. The 2017 AAVA annual self-study from a required reading list.
meeting marks the twelfth year of an event planned Taking the FAAVA examination is a great way to
for practitioners, by practitioners. We know that this refresh and add to your acupuncture knowledge. Passing
conference will be an extraordinary opportunity for the examination is a wonderful way to demonstrate your
members to learn and receive top quality continuing dedication to practicing high quality veterinary acupuncture
education (CE). Your colleagues are in touch with exactly to your clients as well as your colleagues. The examination
what you want to know- and deliver! It is a great place to will take place on March 23, 2017 in Savannah, Georgia
meet experts in your field and network! prior to the 2017 annual meeting.
ABSTRACT
Han JS
Acupuncture and endorphins.
Neurosci Lett. 2004 May 6;361(1-3):258-61.
Acupuncture and electroacupuncture (EA) as complementary and alternative medicine have been accepted worldwide
mainly for the treatment of acute and chronic pain. Studies on the mechanisms of action have revealed that endogenous
opioid peptides in the central nervous system play an essential role in mediating the analgesic effect of EA. Further studies
have shown that different kinds of neuropeptides are released by EA with different frequencies. For example, EA of 2 Hz
accelerates the release of enkephalin, beta-endorphin and endomorphin, while that of 100 Hz selectively increases the
release of dynorphin. A combination of the two frequencies produces a simultaneous release of all four opioid peptides,
resulting in a maximal therapeutic effect. This finding has been verified in clinical studies in patients with various kinds of
chronic pain including low back pain and diabetic neuropathic pain.

AATCVM Updates
Bruce Ferguson DVM, MS President AATCVM
Dr. Ferguson with Suzanne Hughes and Sunny at Upper

Falls in Graveyard Fields in the Pisgah National Forest.
Hello Everyone! I hope that this letter finds you all in next few years will amaze and reward us all for this new,
good spirits and health. I am happy today for a few good significant level of cooperation. Thank yourselves and
reasons. First, I am so happy that the major Veterinary your wise leaders.
Acupuncture organizations throughout the world have I also feel very good with regard to the acceptance
chosen to utilize the American Journal of Traditional of TCVM at the scholastic and practice level throughout
Chinese Veterinary Medicine (AJTCVM) to disseminate our profession. The combined graduate numbers from
information for their organizations as well as a venue for TCVM teaching organizations across the planet is more
publication of quality TCVM research. Please remember than 5,000 registered veterinarians! In the city where
that TCVM includes acupuncture and moxibustion, I currently reside, 75% of the veterinary practices have
Chinese herbal medicine, Tui-Na or Chinese body work, “veterinary acupuncture” advertised as one of the services
and Chinese food therapy. We hope that you feel motivated which their practices offer. Lastly, when clients seek my
to publish results from your successful work with TCVM expertise, they commonly cite stories of great results that
in the journal. If you don’t feel confident in writing or their friends or acquaintances have experienced with their
research, please contact Dr. Dan Hawkins, head of the own animals and TCVM.
AJTCVM research committee, and he can give you some Further, with respect to scholastic acceptance, the
help designing good quality scientific studies and bringing Chi Institute now offers a Masters of Science in TCVM
your results into a publishable form. which is recognized by the accrediting organizations in
So, here we have the International Veterinary the USA. When initially offered, it was predicted that a
Acupuncture Society (IVAS), the American Academy dozen or so veterinarians might seek an advanced degree
of Veterinary Acupuncture (AAVA), the American in TCVM annually. But, to our joy and surprise, over
Association of TCVM (AATCVM), and the World 120 veterinarians have registered for the TCVM Masters
Association of TCVM (WATCVM) all cooperating for degree program, and more are inquiring each and every
the good of ALL veterinary acupuncture across the planet. day. In fact, the first 6 veterinarians have graduated in caps
Cooperation is an absolute necessity for our continued and gowns, and have each done significant research, some
existence and growth. Yet each organization is also of which you will see published soon here in the American
unique and may feel their independence even while in a Journal of TCVM.
cooperative venture such as the AJTCVM. This is very Lastly, I am happy that Hurricane Matthew hit our
important, because it takes fairly large egos to create and home in Saint Augustine on Anastasia Island and caused
maintain such fantastic organizations. And cooperation virtually no damage. Was it the Feng-Shui “cures” placed
is commonly difficult when large egos meet, so we are around our home? Was it my wife’s Pyramids which she
demonstrating highly evolved behavior here my friends, uses to protect our home? Was it the natural buffer of the
and I am quite happy to see it. In fact, the results in the normal, undisturbed ecosystem between our home and the
beach? Whatever the cause, I am so happy that we have of TCVM. She received her Master’s Degree in TCVM
suffered so little. Moreover, those that DID suffer in this (MSTCVM) in 2015 from the Chi Institute and South
massive hurricane have drawn the most positive response China Agricultural College in Guangzhou, China on “The
which humans can show one another. From across the Application of TCVM to Atlantic Bottlenose Dolphins
country, huge convoys of trucks with linesmen and (Tursiops Truncatus). Dr. Clemons-Chevis has been a Chi
electrical restoration workers have made their way here Institute instructor or teaching assistant since 2010 for
and have been working furiously to remove obstructions certification courses of acupuncture in small animals and
and restore power to those without. mixed animals in Reddick, FL, Guangzhou, China, Spain
What a joy it is to see how well humans can cooperate and Costa Rica. In 2015, she was a teaching assistant with
if they really want to do the right thing. My only sadness is the International Veterinary Acupuncture Society (IVAS).
that it often takes a disaster to see this level of cooperation. She has several publications on the use of TCVM in marine
Please, in your own lives, cooperate and help one another mammals and works with marine mammal parks to apply
each and every day! TCVM. She has a TCVM only practice in Bay St. Louis,
MS and the Republic of Panama and serves as Veterinary
Dr. Bruce Ferguson Consultant for the Institute for Marine Mammal Studies,
St. Augustine, Florida Gulfport, MS.
President of AATCVM Dr. Tran was born and raised in Hokkaido, Japan.
After she graduated from Obihiro University of Agriculture
MESSAGE FROM THE AATCVM MEMBER TCVM and Veterinary Medicine (Japan) in 1994, she moved to
CASE DISCUSSION FORUM the United States. She has been practicing small animal
medicine and surgery in Northern Virginia since 1999.
The AATCVM provides case consultations via the She became certified in veterinary acupuncture (CVA)
online Discussion Forum as one of the many benefits in 2007, Tui-na (CVTP) in 2008, and Chinese Herbal
members may utilize. If you are new to the forum and Medicine (CVCH) in 2011 through the Chi Institute. She
wish to learn how to use it, please visit www.aatcvm. is an owner of Pearl Veterinary Holistic Medicine (the
org, login to the ‘Forum’ page, click on the ‘Jump to’ division of Internal Medicine) at the Referral Center of
under ‘Site’, and select ‘Announcement and Instructions Northern Virginia, where she provides Chinese medicine
for using the new Forum’. You may then select the very to companion animals. In her spare time, she works as a
first post titled ‘How to submit new topics and reply to criticalist in the Prince William Emergency Veterinary
existing topics in the new forum’. This post describes Clinic. She is also a teaching assistant at the Chi Institute.
step-by-step instructions on how to submit a new topic to The AATCVM hopes that our members get a chance
the Case Discussion Forum, and how to submit a reply to to contact these wonderful clinicians that have been added
an existing topic. If at any time, you have any questions, to the Case Forum service and we thank Dr Koh for all his
comments or suggestions, please feel free to contact us at: years of service working with members and their difficult
ts@ajtcvm.org. Your feedback and suggestions are very cases as Case Forum moderator.
important to us.
This year the Discussion Forum has two exciting new MESSAGE FROM THE AATCVM RESEARCH
additions. Connie Clemons-Chevis DVM, MSTCVM and COMMITTEE
Rieko Miura Tran BVSc will act as our new moderators
for the forum. Each spring, AATCVM evaluates research proposals
Dr. Clemons-Chevis received her DVM from related to Traditional Chinese Veterinary Medicine
Auburn University College of Veterinary Medicine, (TCVM) from individual researchers and those affiliated
Alabama in 1986. After graduating she started her own with any group or country and awards small grants to
small animal and exotic practice in 1987 in Bay St. those that qualify. The AATCVM has money available
Louis, Mississippi. In 1993, she became the Attending for grants for 2017-2018 and encourages applications for
Veterinarian for Marine Life Oceanarium, Gulfport, MS. research support. All research proposals should be well
After the marine park experienced devastating, Hurricane designed, following the outline listed on the AATCVM
Katrina destruction in 2005, she began studying TCVM website (www.aatcvm.org) illustrating that the applicant
at the Chi Institute in Reddick, FL. Dr. Chevis received understands the problem and proposes a sound scientific
certification as a mixed veterinary acupuncturist (CVA) method of investigation. Completed proposals should be
and veterinary food therapist (CVFT) in 2007. In 2008 submitted to: HawkinsDL@ajtcvm.org. All proposals
she became certified in Chinese Veterinary Herbology must be received by April 1 of the award year and will be
(CVCH) and in 2009 in Veterinary Tui-na, all through the reviewed by the AATCVM Research Committee chaired
Chi Institute. With the fulfillment of all certifications she by Dan Hawkins DVM, MS, DACVS, CVA, IVCA. The
received certification of TCVM practitioner (CTCVMP) committee may request an external review or consultation
in 2013 from the Chi Institute and China National Society by an expert regarding specific issues in the proposal. The

results of the Research Committee evaluation will be sent Development of a long-lasting acupoint stimulation
to the principal investigator by May 1 of the same year. technique utilizing biocompatible hydrogel.
Funding will be awarded to those investigators whose Principal Investigator: Harvey E Ramirez DVM,
projects have been approved by May 30 of the current DACLAM
year. For the next funding cycle, the AATCVM Research Co-Investigators: Pablo R Morales DVM, DACLAM
Committee requests submission of TCVM research and Elizabeth Nunamaker PhD, DVM, DACLAM
proposals as described on the AATCVM website by April Organizations: University of Florida, Gainesville,
1, 2017. Florida (Ramirez, Nunamaker) and The Mannheimer
Foundation, Inc., Homestead, Florida (Morales)
Dan Hawkins DVM, MS, ACVS, CVA, IVCA
2015-2017 AATCVM Funded Research Grants
Effect of oral administration of Yunnan Baiyao on

periprocedural hemorrhage in dogs undergoing nasal
biopsy: A prospective, randomized, double-blinded
controlled study.
Principal Investigator: Shelly Olin DVM, DACVIM
Co-Investigators: Lauren Adelman DVM, Christine
Egger DVM, MVSc, ACVAA and Jennifer Stokes
DVM, DACVIM
Organization: University of Tennessee College of
Veterinary Medicine Veterinary Medical Center
Knoxville, TN

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IVAS Updates
Welcome from the IVAS President 2016-2018: Uwe Petermann DVM, CVA
Dr. Uwe Petermann
Dear members, friends of the IVAS family and all success story that our society has educated and certified
veterinary acupuncture colleagues in the world, thousands of colleagues in veterinary acupuncture and
It is wonderful to see that in a world of concurrence “IVAS certified” is an international recognized brand.
there is a tiny group of very special people - called Another important goal to me is that IVAS established
“veterinary acupuncturists” - who embodies cooperation continuing education events that can be enjoyed by all
even if they belong to different organizations, joining veterinarians. Each year IVAS holds an annual Congress
forces to achieve common goals. in a different location worldwide. This gives veterinarians
For many years my goal was to see that the a chance to combine traveling around the world, seeing
International Veterinary Acupuncture Society would friends and learning about different approaches on
join the AJTCVM, and be part of this excellent peer veterinary acupuncture. Last year IVAS joined forces
reviewed journal in the field of veterinary acupuncture with the American Academy of Veterinary Acupuncture
as a common voice of our passion. I am so happy that (AAVA) and held our annual Congress in Tacoma,
now as the President of IVAS I am able to see this goal on Washington. This year in 2017 IVAS will be joining the
collaboration become a reality. I hope that this will lead to Australian Veterinary Acupuncture Group (AVAG) in
a fruitful development of all of our organizations and all “The Land Down Under!” The 43rd annual IVAS Congress
will prosper in the future. on veterinary acupuncture will be held at the Hilton Cairns
In the year of 1974, when nearly no one knew about in Cairns, Australia August 23-26, 2017. This is where
veterinary acupuncture, a group of veterinarians founded the Great Barrier Reef and the Rainforest meet. We hope
the International Veterinary Acupuncture Society (IVAS) many of you can join us in Australia.
to promote the use of veterinary acupuncture and to IVAS has also been working on adding more
increase education in this modality. I am sure their wish had Continuing Education, another goal of mine was to
been once to see veterinary acupuncture as a commonly have IVAS provide specific topics, which can lead to an
recognized treatment modality. To join our forces is a step advanced certification in that advanced topic. In 2015 we
in that direction, which will finally lead to a complete had two advanced topic courses on “Laser Acupuncture”
recognition of our treatment modality in the veterinary in Europe, one in Milan, Italy and one in Brighton, UK.
medical world, worldwide. I believe it is indeed a great We also had an advanced topic course in “Auricular

Medicine or Ear Acupuncture” in Brussels, Belgium. In considered artists in their daily work, should not stand
the USA we held a joint continuing education event with still; but open their minds for further development of
AAVA in Fort Collins, Colorado. One day addressed acupuncture. As soon as one learns about these modern
“Bi and Wei Syndromes: Advanced Strategies for Those kinds of acupuncture one will realize how close our
Difficult Patients.” The next day was on “Acupuncture modern thinking is with the traditional thinking. This is
and Physical Medicine: Healing with your Hands”. In no wonder, because both systems describe the same nature
2016 we had the first advanced topic course in the USA on but with different knowledge of different times.
“Laser Acupuncture” in Fort Collins, Colorado. This year I wish everyone to enjoy the articles of this issue
IVAS is planning to hold additional continuing education and that you will find new and interesting information
opportunities such as “Food Therapy” in the USA and and hopefully some inspiration for your daily work as
an advanced topic course on “Auricular Medicine/Ear integrative veterinarians.
Acupuncture” in Fort Collins, CO and one on “Gold I wish you a happy, peaceful, healthy, and successful
Bead Implantation” in Europe. We continue to provide New Year in 2017 and hope to meet many of you this
additional opportunities for continuing education. August at our 43rd annual IVAS Congress on veterinary
Since Sunjang the father of veterinary acupuncture acupuncture in Cairns, Australia and – maybe – some of
educated veterinarians in China 3,000 years ago, times you at an IVAS continuing education event in the future!
have greatly changed; as well as the environment, food,
life, work and technique. I think it is reasonable to have Dr. Uwe Petermann
a contribution even in a traditional treatment modality Germany
to modern times. Veterinary acupuncturists, who are President of IVAS
Congress 2016
Group photo of speakers. Gene Tagaban demonstrating the Raven Dance.
The International Veterinary Acupuncture Society the welcome reception, a Native American guest, Gene
(IVAS) and The American Academy of Veterinary Tagaban, blessed the Congress and shared the Raven
Acupuncture (AAVA) joined together June 8-11, 2016 Dance with us. IVAS ends each Congress with a Gala
in Tacoma, Washington for the 42nd annual Congress on dinner including dancing and camaraderie with fellow
Veterinary Acupuncture. IVAS has been hosting an annual colleagues from around the world. Make plans to join us
Congress for 42 years and had attendees representing over for the 43rd annual Congress in Cairns, Australia August
15 countries in Tacoma. 23-26, 2017!
There were a total of 18 speakers including our For more information on how to become a member
Keynote Presenter Dr. Linda Boggie. In addition, there of IVAS and get our Congress Proceedings as a benefit of
were “Poster Presentations” and “HandsOn” canine IVAS membership please visit the IVAS website at www.
and equine wet labs. On Wednesday evening during ivas.org.

Dr. Uwe Petermann’s wet lab.
Dr. Jean Yves Guray’s wet lab.
Dr. Jean Yves Guray’s wet lab. Dr. Linda Boggie leading a wet lab.
Advanced Topic on Laser Acupuncture of the concept of diagnosis by using RAC/VAS pulse
(October 21-23, 2016) control to find active acupoints through automonic reflex
In October IVAS held our first Advanced Topic in activity of vessels. Saturday morning continued the
the US in Fort Collins, CO on “Laser Acupuncture” with lectures with more refinement on the use of acupuncture
Dr. Uwe Petermann. It was a perfect time to have some for chronic musculoskeletal disease and infected wounds
Continuing Education in Colorado, cool crisp mornings using both laser acupuncture and LLLT demonstrating
and warm fall afternoons. We had a full class of 22 their synergism for effective treatment of conditions that
attendees with the furthest attendee coming all the way have had a poor response previously. Saturday afternoon
from Singapore and the second longest distance from combined a hands on lab with canines and “guinea pigs”
Hawaii, not counting the speaker coming from Germany! (i.e. students in the class) to undergo laser diagnosis and
These two and a half days were filled with information and treatment. Sunday it was off to the barn to work with some
hands on learning. equines as well as breaking into small groups to learn the
Friday was a day filled with lectures covering nuts and bolts of operating the lasers with a little RAC
information on types of lasers, the connection between diagnosis on each other! The attendees left with new ways
increased mitochondrial ATP production and laser therapy, to improve treatment in those cases where other treatments
along with scientific studies that support their use and have shown poor results and looking forward to going
effectiveness. Woven into the fabric of how to use laser home to use their new found knowledge.
acupuncture in a host of diseases was the introduction
Canine Patient “Putter”: I love Laser Acupuncture! Dr Petermann demonstrating how to use a laser for RAC
diagnosis on a class volunteer.
Class photo of Laser Acupuncture Course attendees and Dr Petermann

(holding horse) with one of the equine patients.
One of the course participants performing laser

acupuncture on an equine patient.

WATCVM Updates
Bruce Ferguson DVM, MS President WATCVM
I hope that this is a fine day for all of you, and that you president J.F. Kennedy once said “ask not what your
realize how wonderful your lives probably are, especially country can do for you, but rather what you can you do
compared to many others on this planet who are not so for your country”. So I implore my colleagues to “ask not
fortunate. My wife, Suzanne Hughes, and I just returned what TCVM can do for you, but rather what you can do
to our property and home in Saint Augustine Beach after for TCVM”.
Hurricane Matthew hit us directly! And, perhaps by the It is a well-known fact that most organizations in
power of the pyramids which my wife uses to protect the world have only a very small percentage of members
and nourish our home, we returned to a messy yard but who actually contribute their time to Service to support
absolutely no property damage. We are so fortunate. that organization. Let’s see if we can do better than that
Today, as the President of the World Association in the WATCVM! And if I can, with relatively advanced
of Traditional Chinese Veterinary Medicine, I want to Parkinson’s disease, contribute some of my time and
explore two concepts that I believe enrich all of our lives energy in the Service of WATCVM, certainly some of you
and connect us more deeply with this fascinating and can as well.
useful TCVM passion that many call a career. How do our Next, let us briefly explore Cooperation. The word
current careers in TCVM relate to the newly established Cooperation is derived from the Latin “cooperatio” and
WATCVM with respect to the following concepts? French Cooperation. It’s primary definition, which I
First we have “service”. Service is an interesting would like to emphasize is: Active help from a person,
word. It is derived from the Old French “servise”, organization, etc., such as an orderly sharing of space or
meaning “servant” or “slave”. Currently, service has many resources and association for mutual benefit. The antonym
definitions, but let me focus on two including: an act of of Cooperation is competition. The world is full of
being of assistance to someone or something, and to be complexities and more information than any single human
subservient to or employed by an individual or group. It can possibly manage or understand. I believe no single one
is my opinion and strong feeling that we should honor our of us can make a significant impact on the planetary health
world, our relatives, our friends, and our teachers. And and well-Being without having cooperative coworkers
as an extension of our teachers, our TCVM as well. And and colleagues.
honoring them by returning some Service to this medical This is why we develop organizations such as the
system that we love so much. WATCVM. So that through cooperative energies, we can
I would first ask, is it Service that is from the heart; help to bring TCVM into the modern scientific era and
free, unconditional, and unselfish. Many people, perhaps facilitate the standardized teaching and practice of TCVM
more so in these modern, individualistic times, want to throughout the world. There is much work to be done,
know what the world, and especially TCVM, can do and we need the Service as well as Cooperation of our
for them. But I feel the opposite. In fact, the American WATCVM members to accomplish these goals.

In summary, the WATCVM is a SERVICE the members work together as a cooperative group. So,
organization which can only grow to fulfill its goals by please, give some Service to and Cooperate with the
the hands and minds of its members, each giving a little WATCVM for the benefit of ALL TCVM practitioners and
service to the whole organization. The WATCVM is a our patients.
COOPERATIVE endeavor among and between members Have a glorious day!
throughout the world. Such an endeavor only matures Dr. Bruce Ferguson
and shines the light of wisdom and health care when President of WATCVM
18th Annual TCVM Conference Highlights from China

In August, 2016, approximately 84 veterinarians from China along with over 100 veterinarians from 20 different countries
gathered in Beijing, China, for the 18th Annual International Conference co-hosted by the World Association of TCVM, Chi
Institute, Beijing Small Animal Veterinary Association, and China National Society of TCVM. Over the duration of three
days, speakers presented on the topics of clinical application of TCVM schools, behavioral problems (spirit-mind-body),
infections, inflammation and cancer.
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Basic Science Studies
A Randomized, Blinded, Double Controlled Study of the

Effects of Modified Ma Xing Shi Gan Tang on Mortality,
Recovery and Serum Superoxide Dismutase, Glutathione
Peroxidase and Malondialdehyde in Chickens with
Infectious Bronchitis
Yongping Yan DVM, PhD, Xiaodan Wang DVM, PhD, Ruiwen Li DVM, PhD,
Xiu-hui Zhong DVM, PhD
ABSTRACT
Avian infectious bronchitis virus (IBV) causes high mortality and reduced production even in vaccinated birds. The study
hypothesis was that a decoction of a Chinese herbal medicine, modified Ma Xing Shi Gan Tang, in the drinking water would
reduce mortality and improve recovery and antioxidant capabilities in chickens infected with IBV. Two hundred 15-day-old
chickens were randomly divided into 5 groups of 40 each. One group was an untreated control group. Chickens in the other
4 groups were inoculated intranasally with IBV and all developed reduced activity and appetite and respiratory signs within
48 hours. Chickens in 3 groups received 0.6%, 0.4% or 0.2% modified Ma Xing Shi Gan Tang twice daily in the drinking
water for 5 days beginning 48 hours after IBV inoculation. Clinical signs were monitored and deaths and recoveries recorded.
Serum superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) were evaluated on 6
randomly selected chickens from each group, 5 times over a 3-15 day period and the M±SD statistically compared. Mortality
was very significantly (p<0.01) reduced from 42% in the untreated group to 5%, 10% and 27% (0.6%, 0.4% or 0.2% herbs
respectively) and affected birds recovered with no adverse effects from the herbs. Chickens receiving 0.6% or 0.4% modified
Ma Xing Shi Gan Tang had very significantly (p<0.01) higher serum SOD and GSH-Px and lower serum MDA than untreated
IBV chickens. The antioxidant properties of modified Ma Xing Shi Gan Tang make it a novel non-toxic medication for the
treatment and potential prevention of IBV infections of chickens.
Key words: avian, infectious bronchitis, Chinese herbal medicine
(IBV), a member of the family Coronaviridae and genus

ABBREVIATIONS
Coronavirus.1,2 The disease is characterized by clinical
signs of tracheal rales, nasal exudate, coughing, sneezing,
IBV Infectious bronchitis virus
poor weight gain and a decline in egg production and
GSH-Px Glutathione peroxidase
egg shell quality. Additionally birds become susceptible
MDA Malondialdehyde
to secondary pathogens, such as Escherichia coli.3 Avian
M±SD Mean +/- the standard deviation
infectious bronchitis is currently a major problem in China
ROS Reactive oxygen species
and is economically important to the poultry industry due
SOD Superoxide dismutase
to the high mortality, morbidity and production losses
TCVM Traditional Chinese Veterinary Medicine
associated with the disease.4,5 There is no specific treatment
for IBV prevention and control except for modified live or
Avian infectious bronchitis is an acute, highly killed vaccines. In spite of intensive vaccination programs,
contagious, and primarily respiratory infection in frequent outbreaks occur in the field due to the presence
chickens caused by the avian infectious bronchitis virus of different serotypes, generated by point mutations,
From: The Institute of Traditional Chinese Veterinary Medicine
insertions, deletions or RNA recombination of S1 genes.6
Agricultural University of Hebei, Baoding City, China (Yan, Therefore, it is urgent that novel drugs with high efficacy,
Wang, Li, Zhong) and Baoding Vocational and Technical College, low toxicity and extensive availability be developed to
Baoding, China (Yan)

treat and prevent this disease of chickens. Chen, Luo and Wang reported that a modified Ma Xing Shi
There are recent studies of the etiology, epidemiology, Gan Tang ameliorated lung inflammation and inhibited
diagnostic findings and control of avian infectious virus replication significantly in respiratory syncytial
bronchitis, but the oxidant-antioxidant status in chickens virus-infected mice.17
during the course of the disease has not been thoroughly The aim of the study was to find a novel drug with
studied.1 Reactive oxygen radicals have been proposed high efficacy and low toxicity to treat IBV infections
to be involved in the pathogenesis of many diseases, and in chickens. The objective of the current study was to
the elimination of free radicals is usually dependent on evaluate the effectiveness of 3 concentrations of a modified
effective function of the antioxidant defense system.7-9 Ma Xing Shi Gan Tang on mortality, recovery and serum
Antioxidant enzymes, such as superoxide dismutase parameters of oxidative stress in an experimental avian
(SOD) and glutathione peroxidase (GSH-Px) have been infectious bronchitis model. The hypothesis was that
considered to possess beneficial effects against various a decoction of modified Ma Xing Shi Gan Tang in the
diseases that are mediated by oxygen-containing free drinking water of chickens could significantly improve
radicals. The SOD enzyme protects cells from damage recovery and antioxidant parameters in chickens infected
by removing superoxide radicals and malondialdehyde with IBV, compared to untreated infected chicken
(MDA). The GSH-Px enzymes protect the integrity of controls.
cell membrane structure and function. The activity levels
of SOD and GSH-Px can be used as a direct indicator MATERIALS AND METHODS
of oxygen-containing free radical scavenging and the All Chinese herbs were obtained locallya and
strength of the anti-oxidant system. While serum MDA authenticated by the Institute for medicinal plants, China
levels can be used as an index to reflect the extent of cell Academy of Medical Sciences (Beijing, China). The
membrane damage due to attacks by oxygen-containing ingredients of modified Ma Xing Shi Gan Tang consisted
free radicals.9 Evaluation of serum SOD, GSH-Px and of Ma Huang (Ephedra), Shi Gao (Gypsum), Xing Ren
MDA in avian IBV infections before or after experimental (Armeniaca) and Gan Cao (Glycyrrhiza), with Huang
treatments can provide an objective basis for the evaluation Qin (Scutellaria), Ban Lan Gen (Isatidis) and Shan Dou
of novel therapies for treatment and prevention. Gen (Sophora) added.18 The amount and actions of each
Chinese herbal medicines have been used to treat herb are outlined in Table 1. The total weight of the
many diseases of human beings and animals for thousands raw materials was 510 grams (g). Initially Ma Huang
of years in China.10 The Chinese herbal medicine Ma (Ephedra) and Shi Gao (Gypsum) were boiled alone in
Xing Shi Gan Tang, a decoction of Ma Huang (Ephedra), distilled water for 30 minutes (min) before the other herbs
Xing Ren (Armeniaca), Gan Cao (Glycyrrhiza) and Shi were added. Then 3,570 ml (about 7 times the herbal
Gao (Gypsum), was documented in the famous textbook weight) of distilled water was added to cover the entire
Shanghan Lun (Treatise on Febrile Diseases) published herbal mixture and soaked for an hour. The herbal mixture
in the Han Dynasty (about 220AD) in China and is still was then boiled for another hour. The initial herbal liquid
popularly used today in both human and veterinary (first mixture) was separated from the herbal residues and
medicines.10,11 Ma Xing Shi Gan Tang has a modulatory saved. Then 1,530 ml (3 times the herbal weight) was
effect on the immune function of children and experimental added to the herbal residue and boiled again for an hour
animals with acute respiratory tract infections.10-12 Ma after which the herbal liquid was again separated. The first
Xing Shi Gan Tang has also been studied as an effective and second herbal liquids were combined and simmered to
Chinese herbal medicine for treating influenza.13-15 Zhu reconstitute to a final volume of 510 ml. The final herbal
and Liu reported positive therapeutic effects of modified decoction had a concentration of 1 g/ml of modified Ma
Ma Xing Shi Gan Tang on chronic bronchitis in humans.16 Xing Shi Gan Tang and was stored at 4 °C until used, as
Table 1: Ingredients and actions of the Chinese herbal medicine modified Ma Xing Shi Gan Tang
Pin Yin Name English Name Amount (g) Actions18

Ma Huang Ephedra 45 Induces sweat, relieve the Exterior, relieve cough and asthma
Shi Gao Gypsum 135 Clears Lung Heat
Xing Ren Armeniaca 60 Disperse Lung Qi to relieve cough and dyspnea
Gan Cao Glycyrrhiza 45 Harmonize the formula
Huang Qin Scutellaria 90 Clears Heat, dries Dampness
Ban Lan Gen Isatidis 90 Clear Heat from Blood level, Benefit throat
Shan Dou Gen Sophora 45 Clears Lung Damp Heat, reduces cough
g=grams; ingredients decocted to a 1 g/ml solution
previously described.19 sneezing, rattling or nasal discharge) with activity, appetite

Two hundred day-old male Roman chickens were and water consumption returning to normal. The recovery
purchasedb and housed in a climate-controlled room at 36 rate was defined as the ratio of completely recovered
± 1°C and kept under 24-hour (h) lighting at the beginning chickens to the 40 chickens in each group. The total
of the pretrial period. The temperature was gradually effective rate was defined as the number of chickens that
reduced to room temperature and the photoperiod was survived compared to the 40 chickens in the group. The
reduced to a 12 h light and 12 h dark cycle, then kept effective rate of modified Ma Xing Shi Gan Tang at the
constant throughout the experimental period. Chickens 3 different concentrations (Groups III-V) was determined
were fed with a commercial starter dietc. The chicks were and compared for Groups III, IV and V
allowed to acclimate to the environment for 2 weeks and Blood samples were collected from 6 randomly
were 15 days old when the experiment was started. None selected chickens per group at 18, 21, 24, 27 and 30 days
of chickens were vaccinated against IBV. The housing and of age (3-15 days after IBV inoculation and 1-13 days after
handling of the experimental animals were in accordance initiation of modified Ma Xing Shi Gan Tang treatment).
with the guidelines of the Chinese Council for Animal Ten milliliters of blood were removed from the inferior
Care. vena cava and then the chickens were euthanized. Blood
Two hundred 15-day-old chickens were randomly samples were centrifuged at 3,000 rpm for 10 minutes and
divided into 5 groups of 40 each (Table 2). Group I was serum collected. Serum was stored at -70 ˚C until analysis
an untreated control group (no IBV or herbal medicine for SOD, GSH-Px and MDA. Assay kits for SOD, GSH-Px
treatment administered). Group II was an IBV model and MDA were purchasede and analysis procedures were
group (inoculated with IBV, but received no herbal performed according to the manufacturer’s instructions.
medicine treatment). Chickens in Groups III, IV and V Total serum SOD activity was determined by the xanthine
received high, middle and low doses of modified Ma Xing oxidase method. The GSH-Px activity was measured by
Shi Gan Tang respectively. All chickens in Groups II-V the dithio-dinitrobenzoic acid method. The MDA levels
were inoculated intranasally at 15 days of age with 0.3 ml were measured using the thiobarbituric acid (TBA)
IBV M41 strain virus liquidd containing 10-6.25 EID50/0.1ml method.
of IBV-M41. Birds in the herbal treated Groups (Groups III, The mean +/- the standard deviation (M±SD) of the
IV, V) were administered 0.6%, 0.4% or 0.2% modified serum assays of chickens in each group was calculated.
Ma Xing Shi Gan Tang respectively in the drinking water Data from the groups were compared with one-way
twice daily beginning at 17 days of age, 48 h after IBV ANOVA and Chi Square performed with statistical
inoculation and continuing for consecutive 5 days until 21 softwaref. A p-value of less than 0.05 (p<0.05) was
days of age (Table 2). considered significant and p<0.01 was considered very
Clinical signs were recorded daily for 15 days by significant.
trained technicians who were blinded to the treatment of
each group. The number of chickens in each group that RESULTS
died was recorded and the chickens were necropsied. Chickens in Group I (not inoculated with IVB and
The mortality rate of each group was calculated as the receiving no herbal treatment) remained healthy showing
ratio of dead chickens to the initial 40 in the respective no abnormal clinical changes. On clinical examination
group. Recovery of chickens in Groups II-V was defined 48 hours after inoculation with IBV, chickens in Groups
as resolution of major clinical signs (lethargy, reduced II-V became less active, lethargic and began having
appetite, difficulty breathing, lethargy, rales, coughing, difficulty breathing, with rales, coughing, sneezing or
Table 2: Control and Chinese herbal medicine modified Ma Xing Shi Gan Tang treatment group interventions
Number of
Group Number Intervention
Chickens
I 40 Untreated control group; no IBV or modified Ma Xing Shi Gan Tang
IBV model group; intranasal IBV at 15 days of age; no modified Ma Xing Shi Gan
II 40
Tang
Intranasal IBV at 15 days of age and at 17 days of age, 0.6% modified Ma Xing Shi
III 40
Gan Tang in drinking water BID for 5 days
IV 40
V 40
IBV = avian infectious bronchitis virus inoculation; BID= twice daily
rattling with or without nasal discharge. In Group II (IBV, III-V had returned to normal. No adverse side effects were
no herbal medicine) 17/40 (42.5%) chickens died (Table associated with modified Ma Xing Shi Gan Tang treatment
3). On necropsy the mucous membranes of the trachea were observed.
and bronchi were inflamed and swollen. The trachea The number of deaths was 2/40 (5%), 4/40 (10%)
contained varying amounts of serous mucus and caseated and 11/40 (27.5%) respectively for Groups III-V (Table 3,
material. No abnormalities were observed in the Bursa of Figure 1). In Group III (0.6% modified Ma Xing Shi Gan
Fabricius and other organs. The clinical signs of Groups Tang), 35/40 (87.5%) chickens completely recovered. In
III-V, receiving 0.6%, 0.4% or 0.2% modified Ma Xing Shi Group IV (0.4% modified Ma Xing Shi Gan Tang), 33/40
Gan Tang BID beginning 48 hours after IBV inoculation, (82.5%) chickens recovered. In Group V (0.2% modified
began to improve by day 3 of treatment. Group III-V Ma Xing Shi Gan Tang), 26/40 (65%) chickens recovered
chickens regained activity quickly and had improved (Table 3, Figure 1).
appetites. Difficulty breathing, rales and coughing had The mortality in Group II (IBV, no herbal medicine)
lessened by 5 days after completing the 5-day BID herbal was very significantly higher than that in Groups I, III, IV
treatment (22 days of age). By 7 days after completing the and V (p<0.01) (Table 3, Figure 1). In the three modified
herbal treatment (24 days of age), chickens stopped dying Ma Xing Shi Gan Tang Groups, recovery and effective
and respiratory signs continued to improve. By 12 days rates in Groups III and IV were significantly higher than
after completing the herbal treatment, chickens in Groups that of Group V (p<0.05) (Table 3, Figure 1).
Table 3: Therapeutic effect of the Chinese herbal medicine modified Ma Xing Shi Gan Tang on chickens with avian infectious
bronchitis
Groups n Number of Deaths Mortality Rate Number Recovered Recovery Rate Total Effective Rate
I 40 0 0Ac N/A N/A N/A
II 40 17 42.5Ba N/A N/A N/A
III 40 2 5Ac 35 87.5a 95a
IV 40 4 10Abc 33 82.5ab 90ab
V 40 11 27.5ABab 26 65b 72.5b
Group I (no IBV or herbs); Group II (IBV and no herbs); Group III (IBV and 0.6% herbs); Group IV (IBV and 0.4% herbs);
Group V (IBV and 0.2% herbs); IBV = avian infectious bronchitis virus inoculation; M±SD in the same column with different
letters are significantly different, lowercase letters = p<0.05 and capital letters = p<0.01; N/A= not applicable.
100 a
ab
90 a
ab
80
b
70 b
60 Mortality
50 Mortality Rate
Ba
40 Recovery
ABab Recovery Rate

30
Effective Rate
20
Abc
10 Ac
0
0
I II III IV V
Figure 1: Mortality, recovery and efficacy of the Chinese herbal medicine modified Ma Xing Shi Gan Tang on chickens with
infectious bronchitis virus (IBV); Group I was the no IBV or herbs control group (0 affected, 0 deaths); Group II (IBV and no
herbs); Group III (IBV and 0.6% herbs); Group IV (IBV and 0.4% herbs); Group V (IBV and 0.2% herbs); M±SD in the same
column with different letters are significantly different, lowercase letters = p<0.05 and capital letters = p<0.01

The M±SD serum SOD activity in Group I (No IBV treatment) the SOD activities in Groups III and IV (IBV
or herbal medicine) increased with age (Table 4, Figure and 0.6% and 0.4% modified Ma Xing Shi Gan Tang)
2). By 21 days of age (6 days after IBV inoculation), the were significantly higher (p<0.05) than Group V (IBV and
SOD activities in Group II (IBV no herbal medicine) were 0.2% modified Ma Xing Shi Gan Tang) (Table 4, Figure
very significantly lower (p<0.01) than Groups I, III and 2). By 30 days of age (15 days after IBV inoculation and
IV and by 3 days later (24 days old) very significantly 8 days after completion of modified Ma Xing Shi Gan
lower (p<0.01) than all other groups (Table 4, Figure 2). Tang treatment), the SOD activity was significantly higher
By 24 days of age (9 days after IBV inoculation and 2 (p<0.05) in Groups III and IV compared to Groups II and
days after completion of modified Ma Xing Shi Gan Tang V (Table 4, Figure 2).
Table 4: The serum superoxide dismutase (SOD) of controls and chickens with avian infectious bronchitis treated with the
Chinese herbal medicine modified Ma Xing Shi Gan Tang
18 days old 21 days old 24 days old 27 days old 30 days old
Groups n
U/ml U/ml U/ml U/ml U/ml
I 6 280.44±23.31 281.27±12.26ABa 284.85±11.03Aab 291.77±8.56ab 298.95±9.66a
II 6 276.57±30.81 247.84±22.84Bbc 246.45±17.89Bad 266.90±35.19bac 267.18±13.18b

III 6 287.35±29.13 289.28±14.19Ac 290.38±7.52Aa 289.00±10.27Ad 289.562±18.80ab
IV 6 279.06±11.78 277.12±34.54ABab 282.37±29.72ABac 282.65±25.37abc 288.73±25.08ab
V 6 271.87±11.38 246.45±7.92Bc 255.85±4.28ABd 271.04±34.50ba 265.79±16.00b
Group V (IBV and 0.2% herbs); IBV = avian infectious bronchitis virus inoculation; n= number of chickens in each group;
U/ml = unit of enzyme activity per ml; for Groups II-V 18, 21, 24, 27 and 30 days old = 3, 6, 9, 11 and 14 days after IBV
inoculation respectively; for Groups III-V 18, 21, 24, 27 and 30 days old = 1, 4, 7, 10 and 13 days after initiating a BID 5
day course of modified Ma Xing Shi Gan Tang respectively; M±SD in the same column with different letters are significantly
different, lowercase letters = p<0.05 and capital letters = p<0.01.
350
a ABab ABac ab
ab
Ac
ABa Aab Aa
300 b
b
Bbc Bad
ABd
Bc
250
200
150
100
50
0
I II III IV V
Day 18 Day 21 Day 24 Day 27 Day 30
Figure 2: The serum superoxide dismutase (SOD) of controls and chickens with avian infectious bronchitis virus (IBV) treated
with the Chinese herbal medicine modified Ma Xing Shi Gan Tang; Group I (no IBV or herbs); Group II (IBV and no herbs);
Group III (IBV and 0.6% herbs); Group IV (IBV and 0.4% herbs); Group V (IBV and 0.2% herbs; for Groups II-V Day 18,
21, 24, 27 and 30 = age at 3, 6, 9, 11 and 14 days after IBV inoculation respectively; for Groups III-V Day 18, 21, 24, 27 and
30 = age at 1, 4, 7, 10 and 13 days after initiating a BID 5 day course of modified Ma Xing Shi Gan Tang respectively; M±SD
in the same column with different letters are significantly different, lowercase letters = p<0.05 and capital letters = p<0.01.
Like the serum SOD, the serum GSH-Px activity in (15 days after IBV inoculation and 8 days after completion
Group I (No IBV or herbal medicine) also increased with of herbal treatment), the serum GSH-Px activity was
age (Table 5, Figure 3). The Serum GSH-Px activities significantly higher (p<0.05) in Groups III and IV (0.6%
of Group II (IBV no herbal medicine) became very and 0.4% modified Ma Xing Shi Gan Tang respectively)
significantly lower (p<0. 01) than Group I (no IBV or compared to Groups II and V (0.2% modified Ma Xing Shi
herbal medicine) and Groups III and IV (IBV and 0.6% and Gan Tang) (Table 5, Figure 3).
0.4% modified Ma Xing Shi Gan Tang respectively) when Serum MDA levels in Group I (no IBV or herbal
evaluated at 21, 24 and 27 days of age. By 30 days of age medicine) decreased with age (Table 6, Figure 4). From
Table 5: The serum glutathione peroxidase (GSH-Px) of controls and chickens with avian infectious bronchitis treated with
the Chinese herbal medicine modified Ma Xing Shi Gan Tang
Groups n
nU/ml nU/ml nU/ml nU/ml nU/ml
I 6 1154.62±27.61 1323.53±96.50Aab 1358.82±20.79ADab 1365.13±21.14Aab 1397.90±37.36a
II 6 1145.80±72.64 1120.58±83.09Cab 990.76±63.58Eb 976.89±47.19Dab 1156.14±35.21d

III 6 1167.23±84.02 1265.55±22.17ACa 1297.06±108.95ABab 1312.19±72.88ABa 1347.48±34.54b
IV 6 1073.95±44.53 1289.50±54.36ABab 1284.46±75.62ACab 1347.48±2.52Aab 1336.14±23.65b
V 6 1044.96±41.66 1147.06±85.81BCb 1206.30±84.31BCDEab 1222.69±90.61BCb 1124.37±42.18d
nU/ml= nanounit of enzyme activity per milliliter; for Groups II-V 18, 21, 24, 27 and 30 days old = 3, 6, 9, 11 and 14 days after
IBV inoculation respectively; for Groups III-V 18, 21, 24, 27 and 30 days old = 1, 4, 7, 10 and 13 days after initiating a BID 5
day course of modified Ma Xing Shi Gan Tang respectively; M±SD in the same column with different letters are significantly
different, lowercase letters = p<0.05 and capital letters = p<0.01.
1600
Aab a
ABab ABa
1400 ADab Aab b ACab
ABab Aab b
BCDEab Bcb
Aca
Bcb
Cab d
1200 d
Eb
Dab
1000
800
600
400
200
0
I II III IV V
Figure 3: The serum glutathione peroxidase (GSH-Px) of controls and chickens with avian infectious bronchitis virus (IBV)
treated with the Chinese herbal medicine modified Ma Xing Shi Gan Tang; Group I (no IBV or herbs); Group II (IBV and no
herbs); Group III (IBV and 0.6% herbs); Group IV (IBV and 0.4% herbs); Group V (IBV and 0.2% herbs); for Groups II-V
Day 18, 21, 24, 27 and 30 = age at 3, 6, 9, 11 and 14 days after IBV inoculation respectively; for Groups III-V Day 18, 21, 24,
27 and 30 = age at 1, 4, 7, 10 and 13 days after initiating a BID 5 day course of modified Ma Xing Shi Gan Tang respectively;
M±SD in the same column with different letters are significantly different, lowercase letters = p<0.05 and capital letters =
p<0.01.

18 days of age, the serum MDA levels in Group II (IBV II (IBV no herbal medicine) and Group V (0.2% modified
no herbal medicine) started to increase, becoming very Ma Xing Shi Gan Tang) at 21-27 days of age (Table 6,
significantly (p<0.01) higher than that of Group I (No IBV Figure 4). By 30 days of age the serum MDA levels in
or herbal medicine) at 21, 24 and 27 days of age (Table 6, Group III (0.6% modified Ma Xing Shi Gan Tang) were
Figure 4). The serum MDA levels of Group II decreased to still significantly (p<0.05) lower than all other groups.
the same level as Group I by 30 days of age. All chickens
receiving modified Ma Xing Shi Gan Tang had lower DISCUSSION
serum MDA compared to Group II from 21-27 days of All chickens with avian infectious bronchitis
age. The serum MDA levels in Groups III and IV (0.6% receiving some dose of modified Ma Xing Shi Gan Tang
and 0.4% modified Ma Xing Shi Gan Tang respectively) in the drinking water BID daily for 5 days had reduced
were very significantly (p<0.01) lower than that of Groups mortality compared to untreated chickens. Chickens
Table 6: The serum malondialdehyde (MDA) of controls and chickens with avian infectious bronchitis treated with the
Chinese herbal medicine modified Ma Xing Shi Gan Tang
Groups n
nmol/ml nmol/ml nmol/ml nmol/ml nmol/ml
I 6 12.76±0.23 12.39±0.32ABbc 11.99±0.24BDbc 11.92±0.25BCa 10.79±1.12ab
II 6 13.03±0.28 13.26±0.54Aa 15.26±0.23Aa 13.78±0.31Aab 11.48±0.27a
III 6 12.82±0.29 11.28±1.10Bd 10.83±0.34CD 11.04±0.28Cb 10.07±0.52b
IV 6 11.81±0.25a 11.78±0.23Bcd 10.81±1.31CD 11.00±0.90Cb 10.53±0.81ab
V 6 12.62±0.69 11.96±1.02ABb 12.47±0.65Bbc 12.32±0.29Bab 11.32±1.16a
nmol/ml = nanomoles/liter; for Groups II-V 18, 21, 24, 27 and 30 days old = 3, 6, 9, 11 and 14 days after IBV inoculation
respectively; for Groups III-V 18, 21, 24, 27 and 30 days old = 1, 4, 7, 10 and 13 days after initiating a BID 5 day course
of modified Ma Xing Shi Gan Tang respectively; M±SD in the same column with different letters are significantly different,
lowercase letters = p<0.05 and capital letters = p<0.01.
18
16 Aa
Bca
14 ABbc
Aa
ABb Bbc
Bab a
BDbc Bca Bd Bcd CD
ab cb
12 a
CD Aab ab
b
10
8
6
4
2
0
I II III IV V
Figure 4: The serum malondialdehyde (MDA) levels of controls and chickens with avian infectious bronchitis virus (IBV)
treated with the Chinese herbal medicine modified Ma Xing Shi Gan Tang; Group I (no IBV or herbs); Group II (IBV and no
herbs); Group III (IBV and 0.6% herbs); Group IV (IBV and 0.4% herbs); Group V (IBV and 0.2% herbs); for Groups II-V Day
18, 21, 24, 27 and 30 = age at 3, 6, 9, 11 and 14 days after IBV inoculation respectively; for Groups III-V Day 18, 21, 24, 27 and
30 = age at 1, 4, 7, 10 and 13 days after initiating a BID 5 day course of modified Ma Xing Shi Gan Tang respectively; M±SD
in the same column with different letters are significantly different, lowercase letters = p<0.05 and capital letters = p<0.01.
receiving 0.6% and 0.4% modified Ma Xing Shi Gan Tang respiratory pathway, as well as by exogenous exposure
had 5% and 10% mortality compared to 42.5% in the to damaging agents in the environment.27 Excessive
untreated IBV controls. Further 87.5% of chickens in the ROS are neutralized by endogenous antioxidant enzymes
0.6% modified Ma Xing Shi Gan Tang group and 82.5 % such as SOD and GSH-Px, keeping ROS at very low
of chickens in the 0.4% modified Ma Xing Shi Gan Tang concentrations and in a state of dynamic equilibrium. At
group recovered from all clinical signs with no adverse low levels the ROS may be beneficial and promote cell
drug reactions. proliferation, activate leukocyte and phagocytes to kill
The Chinese herbal medicine modified Ma Xing bacteria, reduce inflammation and detoxify, instead of
Shi Gan Tang used in this study consisted of Ma Huang exerting deleterious effects on cells. Oxidative stress occurs
(Ephedra), Shi Gao (Gypsum), Xing Ren (Armeniaca) and when the production of oxidants exceeds the capacity of
Gan Cao (Glycyrrhiza), with Huang Qin (Scutellaria), the antioxidant defense systems of an organism, or when
Ban Lan Gen (Isatidis) and Shan Dou Gen (Sophora) the efficacy of the antioxidant defense system is reduced.
added. From a traditional Chinese veterinary medicine The imbalance of oxidants and antioxidants damages cell
(TCVM) perspective, the overall actions were to clear and mitochondrial membranes through lipid peroxidation
Heat Toxin associated with IBV infection and facilitate causing cellular dysfunction and destruction.28 Studies
the flow of Lung Qi to relieve asthma (Table 1).18 The have shown that oxidative stress plays an important role in
Chinese herbal medicine used in this study was modified the pathogenesis of many diseases in humans and animals,
from the famous ancient formula Ma Xing Shi Gan Tang such as bronchial asthma, hepatitis, hepatic cirrhosis,
which contains only 4 ingredients: Ma Huang (Ephedra), burns, ischemic shock, trypanosomiasis, post-natal stress
Shi Gao (Gypsum), Xing Ren (Armeniaca) and Gan Cao in dairy cows and selenium deficiency disease.29-33
(Glycyrrhiza). Ma Xing Shi Gan Tang has been shown The GSH-Px and SOD enzymes have antioxidative
to be an effective treatment for pneumonia, chronic stress functions and effectively eliminate oxygen-
bronchitis, bronchial asthma and influenza.13,16,20,21 An oral containing free radicals.34 These enzymes are vital for
liquid Chinese herbal medicine modified from Ma Xing maintaining a healthy balance between oxidants and
Shi Gan Tang named Jin Xin significantly ameliorated antioxidants. In the present study, the antioxidant system
lung inflammation in respiratory syncytial virus infected was significantly changed in chickens after avian IBV
mice, and significantly reduced the viral load in the infection. The activities of serum SOD and GSH-Px in
lung tissues.17 Other researchers found that Ma Xing Shi Group II (IBV no herbal treatment) were very significantly
Gan Tang ameliorated lipopolysaccharide-induced lung (p<0.01) lower than that of the unchallenged control group
capillary hyperpermeability and inflammation in rats.22 (Group I no IBV) (Tables 4 and 5; Figures 2 and 3). It
The anti-asthmatic effects of Ma Xing Shi Gan Tang can therefore be concluded that oxygen-containing free
are mainly due to its stimulation of beta2-adrenoceptors on radicals participate in the pathogenesis of IBV infections
bronchial smooth muscle and its anti-inflammatory ability in chickens. During an IBV infection, an increase in the
to inhibit neutrophils entering the airway.11 In TCVM, Ma number of oxygen-containing free radicals and a reduction
Huang (Ephedra) in Ma Xing Shi Gan Tang eliminates of antioxidants produce an imbalance of oxidants and
phlegm, relieves asthma and improves breathing.23,24 The antioxidants. The large number of oxygen-containing
anti-viral effects of Ma Xing Shi Gan Tang and Ma Huang free radicals produced by the infection causes increased
(Ephedra) have been demonstrated.13,17 In another study Ma consumption of antioxidants reducing the serum SOD
Xing Shi Gan Tang reduced the duration of fever in H1N1 and GSH-Px activities. As in other diseases, the excess
virus infections.14 The therapeutic efficacy of Ma Xing Shi free radicals associated with IBV infections resulted in
Gan Tang has been correlated with actions on the toll-like damage to cell, mitochondrial and other biomembranes
receptor 4, proto-oncogene tyrosine-protein kinase (Src), and internal cellular structures.33-34
and nuclear factor NF-κB (NF-κB) signaling pathway.22 Further support for the role of oxidative stress in the
The active component of Ma Huang (Ephedra) dilates the pathogenesis of IBV infections of chickens is the increase
bronchi to improve breathing and stop coughing.16 Huang in serum MDA levels as observed in the current study.
Qin (Scutellariae) in modified Ma Xing Shi Gan Tang Serum MDA is the direct product of lipid peroxidation
has been reported to clear Lung Heat and have antiviral following free radical attacks on unsaturated fatty acids
properties.16,18, 25 Ban Lan Gen (Isatidis) in the modified within cell membranes. The serum MDA levels are an
Ma Xing Shi Gan Tang also has anti-viral activity.26 indicator of lipid peroxidation and indirectly reflect the
Living organisms under aerobic conditions are degree of cell damage.35 In the current study, the serum
continuously exposed to reactive oxygen species (ROS), MDA level in Group II (IBV no herbal treatment) was
which includes oxygen molecules (O2), superoxide very significantly (p<0.01) higher at day 24 and 27 days of
anion radicals (O2−), hydroxyl free radicals (OH), singlet age than the unchallenged control group (Group I no IBV)
oxygen (1O2), and hydrogen peroxide (H2O2). The ROS (Table 6, Figure 4). The conclusion for these results are
are generated by imbalanced endogenous reduction of that the oxygen-containing free radical reaction increased
oxygen by cellular enzymes or in the mitochondrial 9-12 days after IBV inoculation, resulting cell damage

from lipid peroxidation, as occurs in other diseases.36,37 Acknowledgments

The lipid peroxidation was reflected in the elevated serum This study was financially supported by the
MDA levels. Department of Education Hebei Province (No.
It is believed that the ingredients of Chinese herbal ZC2016147).
medicines have synergistic effects.18,38 In a study of
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Safety Evaluation of Modified Si Jun Zi Tang,

Assessment of Controlled and Randomized, Acute
and Subchronic Mouse Oral Toxicity Studies
Shuang Ma DVM, MS, Aituan Ma DVM, MS, PhD
ABSTRACT
In order to evaluate the clinical safety of a modification of the classical formula Si Jun Zi Tang (Wei Qi Booster®a (WQB)),
acute and subchronic mouse oral toxicity tests were conducted. An acute oral 7-day mouse study using 80 Kunming (KM)
mice randomly divided into a saline control and 7 dosages of WQB (range 2.0 -15.06 g/kg BW) was conducted to determine
LD50. No mortality or adverse effects were observed in any parameter tested which included food consumption, clinical
toxicity, body weights, organ/body weight ratios (liver and kidney) and histologic morphology of the liver, spleen, lung,
kidney and duodenum. To determine the maximum tolerated dose (MTD) of WQB, 30.12 g/ kg BW was given twice daily
for 7 days. No mortality or adverse effects were observed in any parameter tested in this study which included all parameters
evaluated in the LD50 study along with hematology. In the subchronic oral study, 15 mice were divided into 3 groups with
the control group fed normal chow diet and each of the other two groups either receiving normal chow with 1% WQB or
levamisole hydrochloride 0.01 g/kg BW (positive control) for 90 days. No mortality or adverse effects were observed in the
1% WQB in any parameter tested which included all parameters evaluated in the acute studies and with clinical chemistry
replacing hematology. Results of these preclinical safety studies conducted with WQB at the highest dose tested which is
30.12 g/kg BW for 7 days and 1% of food consumed for 90 days demonstrated no toxicity.
Key words: Si Jun Zi Tang, Wei Qi Booster, LD50, MTD, pre-clinical safety testing, hematology; biological
indexes; immunity indexes, mouse, Chinese herbal medicine
ABBREVIATIONS Herbal medicinal products and supplements have

increased exponentially over the past three decades
ALT Alanine aminotransferase and are now used by more than 80% of the worldwide
ALB Albumin population.1 With the increased use of these therapeutic
AST Aspartate aminotransferase agents, the safety of Chinese herbal medicine has drawn
BUN Blood urea nitrogen more attention recently from both domestic and abroad.2
CHOL Cholesterol Establishing the safety of therapeutic compounds through
Crea Creatinine toxicity studies is an active stage of research for western
GLU Glucose pharmaceuticals and is a requirement for approval under
HGB Hemoglobin government regulations that approve the release of new
LD50 Median Lethal Dose 50 drugs. In these studies, safety data are collected with the
LYM Lymphocyte main goal to assess a product’s safety profile. Studies of
MID Leukocytes (neutrophil, eosinophil, basophil) a therapeutic agent’s toxicity include identification of
organs that may be targeted by the compound; as well
MTD Maximum tolerated dose
as any long term adverse effects. These studies should
RBC Red blood cell
mimic the planned route, duration and schedule of drug
TG Triglyceride
administration as closely as possible.3 These non-clinical
UA Uric acid
studies may include acute toxicity (7 or 14 day studies),
WBC White Blood Cell
subacute (28-day study), subchronic (90 days or greater)
WQB Wei Qi Booster
and chronic (1 year) studies usually in a rodent species.
From: College of Traditional Chinese Veterinary Medicine, Body weights, clinical signs of toxicity, food consumption,
Agricultural University of Hebei, Baoding 071000 China (S. Ma, clinical pathology, histopathology, organ weights and
A, Ma)

behavioral changes are usually evaluated in these studies demonstrated anti-neoplastic effects as well as anti-
along with mortality. inflammatory properties.12,13 In addition, the combination
A brief review of the literature reveals there is some of Huang Qi (Astragalus) and Dang Gui (Angelica) has
safety testing of Chinese herbal medicines similar to been shown to stimulate erythropoetin production in
pharmaceuticals being performed with both single herbs cultured kidney cells which therapeutically benefits the
and herbal formulas. Safety evaluation of Xiang Gu treatment of anemia.14
(Shitake) was performed by Gao et al in a 14-day study in Although WQB has demonstrated numerous
mice containing 3 dose groups along with a saline control. benefits when administered to animals both in clinical
There were no adverse events recorded and a complete and pharmacological studies, its safety in non-clinical
blood count and serum biochemical tests of liver and toxicity testing remains unknown. The objective of
kidney were all within normal range.4 The median lethal the present study was to evaluate the safety of WQB in
dose (LD50) and the maximum tolerated dose (MTD) of a acute and subchronic oral rodent studies by the collection
formula containing Huang Lian (Rhizoma coptidis) and Ku of data on body and organ weight changes, histologic
Shen (Sophora flavescens) was tested in a mouse model. changes of selected organs and clinical pathology changes
These animals were given 40 g/kg BW of the test herbs (hematology, clinical chemistry) in order to provide
three times a day for 7 days without mortality or adverse guidance for safe use of this Chinese herbal medicine in
effects on body weight, food and water consumption or clinical applications. The aim of these studies was to set an
clinical signs. The LD50 was determined to be greater than MTD (maximum tolerated dose), and LD50 (median lethal
40g/kg BW and the MTD greater than 120 g/kg BW. The dose) for WQB and identify any long term adverse effects
authors concluded that the traditional Chinese medicines (subchronic exposure) from oral administration of the
had very low toxicity or no toxicity and were safe to use compound. The hypothesis for this group of experimental
in the clinic.5 Kim and his group investigated the safety studies was that WQB would demonstrate very low or no
of Ren Shen Bai Du San (Insampaedok-san water extract, toxicity in these studies.
ISSE) in mice, when receiving 5g/kg BW once daily for
15 days. Again, no mortality or adverse effects occurred MATERIALS AND METHODS
and ISSE was considered safe for consumption.6 The acute Safety assessment of WQB was conducted through
toxicity of the herbal formula Jin Weng Zhi Li Ke Li which a series of 3 safety tests which included 2 acute toxicity
contains several herbs including Huang Qi (Astragalus tests (LD50 and MTD) and 1 subchronic assay. The dosage
membranaceus) and Jin Yin Hua (Lonicera japanica and experimental procedures used in the LD50 and MTD
thunb) was evaluated in mice in a 7-day study. The results studies were consistent with the government legislation
showed the LD50 was greater than 20 g/ kg BW with no “Evaluation Procedures and Methods on Health Food
adverse effects in study parameters evaluated.7 In a 13- Safety Toxicology” and “Preclinical Safety Evaluation
week subchronic oral toxicity study of Liu Wei Di Huang and Experiment” issued by the Chinese Ministry of
Tang, the results showed that all data measured which Health.15,16,17 Consistent with the regulatory requirements,
included mortality, clinical signs, body weight changes, calculation of doses for evaluating compound safety in the
food and water consumption, ophthalmologic findings, LD50 study included multiples of 1.4X for 7 dosages. If no
urinalysis, hematological and biochemical parameters, adverse effects occurred at the highest dose level in this
gross necropsy findings, organ weights, and histological study, then the MTD study dose would be set at double the
examination of organs, were within normal limits.8 highest LD50 dose. Dose determination for the subchronic
A modification of the classical formula Si Jun Zi study was based on a previous study conducted with
Tang (Wei Qi Booster®a) has been used in multiple species chickens and was set at 1% WQB added into the normal
of domestic animals with success to tonify Wei Qi, Qi chow for 90 days.10
and Blood, and clear Heat-Toxins.9,10 This Chinese herbal All animals for these studies were sourced from the
medicine consists of Dang Shen (Codonopsis pilosula), Laboratory Animal Center of Hebel Medical University.
Huang Qi (Astragalus membranaceus), Wu Yao (Lindera The mice were housed in polypropylene cages with
aggregata), Chen Pi (Citrus reticulata), Dang Gui (Angelica cellulose fiber chip bedding at a temperature of 25ºC and
sinensis), Xuan Shen (Scrophularia ningpoensis), Bai 30-40% humidity along with a 14-hour light and 10-hour
Hua She She Cao (Hedyotis diffusa) and Ban Zhi Lian dark cycle. All animals were given free access to rodent
(Scutellaria barbata). Numerous pharmacological studies feed and water in glass bottles with rubber stoppers for an
have shown that this group of herbs can enhance body adaptive period of 7 days. All animal care and use was in
immunity and provide antitumor effects (references?).11-14 accordance with the guidelines of the Chinese Council for
Liu and his colleagues demonstrated that Huang Qi Animal Care.
saponins, one of the main ingredients of Huang Qi The first acute toxicity test performed was to
(Astragalus membranaceus), inhibits tumor growth in the establish an LD50 for WQB. This study contained 80,
hepatoma 22 (H22) model and sarcoma 180 (S180) implants 8-week-old adult mice with an average weight of 30±2
in mice.11 Bai Hua She She Cao (Oldenlandia) has also g. After the adaptive period, the mice were randomly

divided via a random number table into 8 groups with humanely euthanized as previously described and blood
10 mice in each group. Mice in the control group were was collected for clinical chemistry evaluation. Serum
gavaged with 0.4 ml normal saline, while mice in groups was obtained from the blood samples by centrifugation
1-7 received the same amount of saline mixed with 2.0 g, at 3,000 rpm for 10 min. The level of triglyceride (TG),
2.80 g, 3.92 g, 5.49 g, 7.68 g, 10.76 g and 15.06 g WQB/ cholesterol (CHOL), glucose (GLU), uric acid (UA),
kg BW twice daily for 7 days respectively. The animals creatinine (Crea), alanine aminotransferase (ALT), blood
received daily evaluation of food consumption, clinical urea nitrogen (BUN), total protein (TP), albumin (ALB)
signs of toxicity and monitoring for mortality. After 1 and aspartate transaminase (AST) were measuredf. Final
week of treatment, the mice were humanely euthanized by body weight and organ weights (liver, kidney) were
ketamine/xylazine overdose and blood was collected for recorded along with collection of the liver, spleen, lung,
hematology. Parameters evaluated included intermediate kidney and duodenum for histopathologic evaluation. The
cells (eosinophil, neutrophil, basophil leukocytes) tissues were fixed in Bouin’s solution and processed for
(MID), lymphocytes (LYM), red blood cells (RBC) histology by dehydration in a graded ethanol series. They
and hemoglobin (HGB) with a commercial hematology were then embedded in paraffin and serial sections (4-5 um
analyzerb. Final body weights were recorded along with in thickness) were stained with a standard hematoxylin-
immediate collection of liver, spleen, lung, kidney and eosin (H&E) stain for evaluation by light microscopy.
duodenum from the control and high dose groups. The Data were expressed as means plus or minus the
organs were weighed and then placed in Bouin’s solution standard deviation (mean ± SD). Differences between
for histopathological evaluation. groups were evaluated by one-way analysis of variance
The second acute toxicity test was to establish the (ANOVA) using SPSS software package for Windowsg. In
MTD for WQB. This is the highest dose of an agent that all tests, statistical significance was considered p<0.05 (*)
can be administered without causing excessive toxicity or significant, and p<0.01 (**) very significant.
decreased survival. This study contained ten 8-week-old
mice, with an average weight of 30±2 g. It was conducted RESULTS
as part of the LD50 study, therefore, the control animal There was no mortality in the acute studies (MTD,
group was shared. The 10 study animals received 30.12 LD50) in any study group over the 7-day observation
g/kg BW of WQB twice daily for 7 days. All animals period. In addition, no meaningful differences were seen
were given free access to food and water after gastric in food consumption and clinical signs of toxicity when
infusion. The animals received daily evaluation of food treated groups were compared to the control group. The
consumption, clinical signs of toxicity and monitoring ratio of liver and kidney organ weights to final body
for mortality. After 1 week of treatment, the mice were weight did not show any statistically significant effect in
humanely euthanized as previously described and blood the WQB dose groups of the LD50 study, including the
was collected for hematology (MID, LYM, RBC, HGB) 30.12g group in the MTD study (p>0.05) (Table 1). In
with evaluation performed by a commercial hematology addition, there were no statistically significant findings in
analyzerb. In addition, final body weight and organ weights any hematology parameter evaluated in either acute study
(liver, kidney) were recordedc. Protocol organs which (Table 2).
included liver, spleen, lung, kidney and duodenum were In both acute studies, histological examination of
immediately collected and fixed in Bouin’s solution for liver sections from the 15.06 g WQB group in the LD50
histological evaluationd. Both acute studies were repeated study (Figure 1 B) and the 30.12g WQB group in the MTD
twice to verify results obtained. study (Figure 1 C) demonstrated normal hepatocellular
Subchronic toxicity studies (90-day or 13 weeks) morphology with no evidence of hepatocyte hypertrophy
are conducted to evaluate the safety of long-term or cytoplasmic vacuolation (lipidosis) when compared
administration of drugs or other therapeutic products. To with their control groups (Figure 1 A). Examination of
evaluate the long-term safety of WQB, a 90-day study white pulp in the splenic sections demonstrated normal
was conducted in mice. The study contained 45 healthy size and normal lymphocyte populations and volume in
8-week-old mice randomly divided into 3 groups of 15 both the 15.06 g and 30.12 g WQB groups (Figure.1 E,
animals. The control group was fed normal chow diet with Figure.1 F) when compared to the control group (Figure 1
each of the other two groups either receiving normal chow D). Both WQB groups (Figure 1 H, Figure 1 I) had normal
combined with 1% WQB or levamisole hydrochloridee lung morphology when compared to controls (Figure 1 G)
0.01 g/kg BW. Levamisole hydrochloride was used as a characterized by normal alveoli with no interstitial fibrosis
positive control due to its confirmed ability to enhance or hemorrhage. Kidney sections in the treated groups
body immunity and used as comparison of immune (Figure 1 K/L) were examined for inflammatory infiltrates,
enhancement that may occur subsequent to dosing with glomerular changes, fibrosis, renal tubule cell injury and
WQB. Mice were weighed every 2 weeks and received urothelium cell loss and were not different from the control
daily evaluation for signs of clinical toxicity, food group (Figure 1 J). Examination of duodenal sections in
consumption and mortality. After 90 days, the mice were the 15.06 g WQB group from the LD50 study (Figure 1 N)

Table 1: Acute toxicity studies; Effect of WQB on organ to final body weight ratio of liver and kidney (n=10)
WQB Dose (g per Liver weight to body Kidney weight to

Group P-value P-value
kg BW) weight ratio body weight ratio
Control - 4.17±0.11 1.28±0.10
1 2.00 4.25±0.44 0.81 1.21±0.02 0.64
2 2.80 4.50±0.19 0.32 1.15±0.07 0.39
3 3.92 4.44±0.26 0.40 1.27±0.11 0.95
4 5.49 4.22±0.18 0.88 1.35±0.13 0.69
5 7.689 4.45±0.25 0.39 1.37±0.06 0.59
6 10.76 4.75±0.18 0.09 1.40±0.12 0.47
7 15.06 4.21±0.08 0.90 1.27±0.10 0.91
MTD 30.12 4.55±0.22 0.25 1.28±0.20 0.98
In all Tables, statistically significant (p<0.05), is denoted with (*) and very significant (p<0.01), with (**); statistics calculated
by ANOVA (Tables 1-5).
Table 2: Acute toxicity studies; Effect of WQB on hematological parameters (n=10)
Dose RBC
Group MID(%) P-value LYM(%) P-value P-value HGB(g/L) P-value
(g per kg BW) (1012/L)
Control
- 7.24±0.64 26.46±0.66 4.78±0.27 134.3±5.76
Group
1 2.00 6.97±0.50 0.61 24.58±1.26 0.26 4.56±0.30 0.54 128.2±2.08 0.37
2 2.80 7.42±0.41 0.75 27.14±2.00 0.67 4.35±0.24 0.24 133.7±5.37 0.93
3 3.92 6.36±0.44 0.11 25.55±1.90 0.58 5.06±0.26 0.44 136.0±6.39 0.79
4 5.49 7.60±0.34 0.51 24.01±0.55 0.14 4.73±0.23 0.89 131.4±6.50 0.67
5 7.68 7.20±0.30 0.94 26.45±1.16 0.99 4.96±0.29 0.62 127.8±3.01 0.34
6 10.76 6.82±0.03 0.44 25.68±0.33 0.63 4.86±0.23 0.82 125.7±5.32 0.21
7 15.06 6.74±0.24 0.36 25.65±0.41 0.62 5.13±0.26 0.35 131.0±2.99 0.63
MTD
30.12 6.70±0.09 0.32 25.27±0.35 0.47 4.72±0.19 0.87 124.2±1.65 0.14
Group
*(p<0.05) significant; **(p<0.01)very significant
and the MTD study (Figure 1 O) when compared with the the levamisole group (P-value=0.02) and 1% WQB group
control group (Figure 1 M) were normal with no villus (P-value=0.05) (Table 4). There were minor increases
blunting or crypt hyperplasia. and decreases in various clinical chemistry parameters
In the subchronic 90-day study, all mice grew well when the 1% WQB group was compared to the control
with normal food consumption along with no mortality group, however, none of the differences were statistically
or signs of clinical toxicity. There was no statistically significant (Table 5) and all chemistry values were within
significant body weight effect (p>0.05) in any of the the study reference range.
treatment groups at any study time point. All animals Histopathology of protocol organs in the subchronic
gained weight equally throughout the study (Table 3) study included liver, spleen, lung, kidney and duodenum.
when body weights at various study time points were Liver sections were examined for toxicologic changes
compared to starting weights during week 1. In addition, such as abnormalities of the artery, vein and bile ducts in
there was no statistically significant difference in the the portal triad, hepatocyte hypertrophy, areas of necrosis
kidney/body weight ratio when the treatment groups were and numbers of stellate cells and reticulum fibers in the
compared to the control group. A statistically significant perisinusoidal spaces (Figures 2 A/B). The white and red
difference occurred in the liver/body weight ratios both in pulp were evaluated in the splenic sections for volume of

Figure 1: Effect of WQB on liver, spleen, lung, kidney and duodenum histologic morphology in acute toxicity studies (n=10)
Photomicrographs of liver: A-C; spleen: D-F; lung: G-I; kidney: J-L; duodenum: M-O
Control Group: A, D, G, J, M; Dose Group (15.06 g/kg BW): B, E, H, K, N; MTD Group (30.12 g/ kg BW): C, F, I, L, O;
Hepatocyte morphology is normal (arrow) in all WQB dosed groups with no evidence of hepatocellular cytoplasmic vacuo-
lation or lipidosis. There is no difference between the volume of white pulp with lymphocyte numbers and germinal centers
in the spleen (arrow) when 15.06 g/kg BW WQB group and MTD group is compared with the control group. Lung histologic
morphology in the 15.06 g/kg BW WQB group and in the MTD group demonstrates normal lung alveoli (arrow), no intersti-
tial fibrosis or hemorrhage when compared to control mice. Kidney glomerular morphology (arrow) along with renal tubular
epithelium in 15.06 g/kg BW WQB group and in MTD group is normal and shows no evidence of hyperplasia, vascular chang-
es or cellular injury when compared to the control group. The duodenum in the 15.06 g/kg BW WQB group and in MTD group
demonstrates normal villous height and cellular morphology (arrow) with no evidence of villus blunting, crypt hyperplasia or
dilatation when compared with the control group. Black bar in right corner of photomicrographs represents 100 μm.
Table 3: Subchronic study; Effect of WQB on body weight (n=15)
Time Control group Levamisole group P-value 1%WQB group P-value
1 week 24.14±0.70 25.12±0.60 0.28 25.02±0.53 0.33

3 weeks 32.58±0.25 31.54±0.38 0.11 32.10±0.59 0.45
5 weeks 36.06±0.76 36.90±0.81 0.39 37.16±0.34 0.27
7 weeks 43.85±0.35 42.90±0.63 0.29 44.58±0.77 0.42
9 weeks 42.90±0.97 43.64±1.26 0.65 45.02±1.09 0.21
11 weeks 44.57±0.27 44.44±0.15 0.72 44.99±0.34 0.28
13 weeks 44.21±0.47 44.25±0.20 0.93 45.16±0.39 0.10
*(p<0.05) significant; ** (p<0.01) very significant
Table 4: Subchronic Toxicity Study; Effect of WQB on organ to final body weight ratio (n=15)
Indexes Control group Levamisole group P-value 1%WQB group P-value
Liver weight to body weight ratio 3.17±0.09 3.39±0.04* 0.02 3.36±0.04* 0.05
Kidney weight to body weight ratio 1.24±0.08 1.38±0.10 0.31 1.36±0.09 0.35
Table 5: Subchronic Toxicity Study; Effect of WQB on clinical chemistry parameters in mice (n=15)
Indexes Control group Levamisole group P-value 1% WQB group P-value

CHOL (mmol/L) 3.54±0.09 3.52±0.04 0.79 3.48±0.07 0.56
TG
2.10±0.14 2.04±0.17 0.75 1.87±0.04 0.25
(mmol/L)
Crea
32.80±0.97 33.22±0.38 0.68 34.00±0.65 0.25
(umol/L)
Glu
5.62±0.45 5.35±0.24 0.63 5.70±0.45 0.89
(mmol/L)
AST
38.00±2.28 38.55±2.19 0.86 40.48±1.90 0.43
(U/L)
ALT
78.40±5.56 78.60±2.80 0.97 82.20±2.73 0.51
(U/L)
TP
52.80±3.23 52.04±2.93 0.85 54.63±1.71 0.64
(g/L)
ALB
32.80±0.37 32.56±0.50 0.66 32.73±0.18 0.90
(g/L)
BUN
9.15±0.58 8.71±0.46 0.50 9.44±0.20 0.65
(mmol/L)
UA
70.49±0.59 71.64±1.63 0.45 70.32±0.54 0.91
(mg/dl)

Figure 2: Effect of 1% WQB in diet on liver, spleen, lung, kidney and duodenum histologic morphology in subchronic
toxicity study (n=15)
Photomicrographs of: Liver A-B: Spleen C-D; Lung E-F; Kidney G-H; Duodenum I-J.
Control Group A, C, E, G, I; 1% WQB Group B, D, F, H, J. The hepatocytes and portal triads (arrow) in 1% WQB and
controls demonstrate normal morphology along with rare observation of stellate cells and reticulum fibers in perisinusoidal
spaces. Splenic red and white pulp (arrow) in 1% WQB and controls was filled with appropriate numbers of red blood
cells and lymphocytes, respectively, with normal germinal centers in the white pulp and no inflammatory infiltrate.
The histologic morphology of lung in both 1% WQB and controls was normal with clear alveolar lumens (arrow), no
congestion, edema, inflammation or fibrosis noted in any sections. The renal stroma, including glomeruli (arrow) in the
1% WQB group was normal and no inflammatory cells, fibrosis or renal tubule cell loss was noted. The morphology of the
intestinal mucosa in the 1% WQB group was normal with no pathological changes of congestion, edema, inflammatory
cell infiltration, erosion or ulcers noted. Black bar in right corner of photomicrographs represents 100 μm.
erythrocytes, size of the germinal centers and increased subchronic study over its 90-day observation period. In
inflammatory infiltrate of the interstitium (Figures 2 C/D). all studies, no meaningful differences were seen in food
Lung section morphology included evaluation for edema, consumption and clinical signs of toxicity when treated
inflammation and fibrosis (Figure 2 E/F) while kidney groups were compared to the control group. The ratio of
sections were examined for inflammatory infiltrates, liver and kidney organ weights to body weight did not
fibrosis, renal tubule cell injury and urothelium cell loss show any statistically significant effects in any acute study
(Figure 2 G/H). The villous morphology of duodenal WQB dose group including the very high dose of 30.12g/
mucosa was assessed for histologic changes such as kg BW group in the MTD study (p>0.05). Histopathology
edema, inflammatory cell infiltrates, erosions and ulcers of all other organs collected in the WQB groups in the
(Figure 2 I/J). All were found to be within normal limits. acute and subchronic studies were within normal limits
for that organ. In addition, both hematology and clinical
DISCUSSION chemistry parameters were unaffected in animals dosed
It is a scientific and regulatory requirement that with WQB in all safety studies.
before any potential new human and some veterinary A statistically significant increase occurred when
drugs can be administered to patients in a clinic situation, comparing control liver/final body weight ratio and
the toxicity of the compounds should be investigated in both treatment groups in the subchronic toxicity study.
order to define safe doses. Safety tests, also referred to as This was considered a non-adverse finding due to the
preclinical safety testing, are intended to reveal potential following: there was no histologic abnormalities including
toxicity associated with a prospective medicine. Careful hepatocellular hypertrophy, clinical chemistry values were
consideration of the doses used in preclinical studies is normal and there was no dose response increase in the
necessary to fulfill the need to identify target organs liver/body weight ratios in the acute studies. Most likely
that may experience excessive toxicity and usually are this finding is secondary to one control group animal with
identified by setting low, intermediate and high doses, plus a low liver weight (outlier) which caused an artefactual
a control group. This span of doses will usually cover from lowering of the group’s liver organ weight ratio results.
no effect to adverse effects or until limited by volume or According to the experimental design and statistical
limit dose (highest dose that should be used in the absence basis of drug evaluation, a Chinese herbal medicine can be
of demonstrable toxicity). considered safe if there is no mortality or adverse clinical
Establishing the safety of Chinese herbal medicines signs in animals by gavage at a maximum tolerated
as they experience increased use in clinic settings is oral dose of 10 g/kg BW.21 There was no adverse effect
becoming more common. Two examples of Chinese herbal observed at the highest dose level tested (30.12 g/kg BW)
medicines used as veterinary therapeutics after preclinical in the acute studies to determine LD50 and MTD. Since
safety testing demonstrated their oral safety include the MTD of WQB was 30.12g/kg body weight (3 times
Di Weng Ke Li composed of Bai Tou Weng (Pulsatillae higher than 10 g/kg BW), 0.14 g of WQB is considered
Radixand) and Huang Lian (Coptidis Rhizoma) for piglets acutely nontoxic to a 50 g mouse. When considering the
and Hu Pi Su (Quercetin) for use as a feed additive in recommended therapeutic dose of WQB for different
animal production. These medicines, similar to the present species, 1.0 mg/kg for humans, 1.87 mg/kg for dogs or
study, had no deleterious effects, including no reduction 9.1 mg/kg for mice given twice daily; the maximum dose
in body weights when tested at the highest dose level tested (30.12g/kg BW) indicates a very large acute margin
possible.18,19,20 of safety (approximately 300X depending on species)
In the current experimental study, a modification of when using this Chinese herbal medicine.
the classical formula Si Jun Zi Tang (Wei Qi Booster®a), In the subchronic study, when evaluating long term
which is a veterinary specific Chinese herbal medicine, exposure, 1% WQB was added into the normal feed of
was evaluated for oral safety by using both acute and experimental animals. With the study mice consuming
subchronic mouse studies modeled after preclinical safety approximately 6-8g of feed per 30-40g BW/day, the
studies used to evaluate new pharmaceuticals. Safety study mice received an approximate dose of 2g/kg or
assessment of WQB was conducted through a series 2000 mg/kg BW of WQB per day for 90 days. When
of 3 safety tests which included 2 acute toxicity tests considering the recommended therapeutic dose of WQB
(LD50 and MTD) and 1 subchronic assay. The dosage for different species, the maximum dose found to have no
and experimental procedures used in the design of these adverse effects (2000 mg/kg BW) again indicates a large
studies were consistent with government legislation chronic exposure margin of safety of approximately 15X
“Evaluation Procedures and Methods on Health Food (depending on species).
Safety Toxicology” and “Preclinical Safety Evaluation Results of this group of safety studies in mice
and Experiment” issued by the Chinese Ministry of with modified Si Jun Zi Tang (Wei Qi Booster®a)
Health.15,16,17 clearly demonstrated no acute or subchronic toxicity to
There was no mortality in the acute studies (LD50, experimental animals in any study . It may be concluded
MTD) over the 7-day observation period or in the that WQB is a safe Chinese herbal medicine, even at

the highest dosage (30.12g/kg BW) for one week and 8. Ha HY, Lee JK, Lee HY et al. Safety evaluation
also long term usage at 1% of feed or 2000 mg/kg BW. of yukmijihwang-tang: assessment of acute and
The hypothesis was therefore accepted that WQB would subchronic toxicity in rats. Evid Based Complement
demonstrate no toxicity in experimental animals under the Alternat Med 2011; doi:10.1155/2011/672136
conditions of these studies. In conclusion, these studies 9. Patricia B. Acupuncture and Chinese Herbal Therapy
provide evidence of the safety of this Chinese herbal for Acute Upper Respiratory Disease in a Geriatric
medicine and support the basis of its safe use in clinical Quarter Horse. AJTCVM 2009;4:52-57.
practice. 10. Li S. Influence of WQB on the immune response to
Newcastle disease vaccine in chickens [D]. Hebei,
ACKNOWLEDGEMENTS China, Agricultural University of Hebei 2013; 5.
This work was financially supported by the Science and 11. Liu MH, Ren MP, Chen JP et al. Study on antitumor
Technology Development Fund of Agricultural University activity of Astragalus saponins. Pharmacology and
of Hebei (NO.1509012) Clinics of Chinese Materia Medica 2009; 25:68-70.
12. Wang Q. Studies on anti-tumor and anti-inflammatory
Footnotes experiment of fujian hedyotis diffusa willd [D].
a. Wei Qi Booster, Dr. Xie’s Jing Tang Herbal Inc, Fujian, China, Fujian University of Traditional
Reddick, FL USA Chinese Medicine 2013; 1.
b. KM-23vet, Komeic company, Shanghai, China 13. Mao Y, Xu F, Xu XJ et al. Research progress on
c. Sartorius, Germany chemic constituent and anti-tumor activity of

d. Huangxing Science and Technology Co. LTD, oldenlandia diffusa willd. Modern Preventive
Beijing, China Medicine 2015;42:3128-3132.
e. Levamisole hydrochloride, Jinpai Pharmaceutical, 14. Zuo ZC, Gan M, Cui HM et al. Effects of Chinese
Xuzhou, China herbal compound on the blood physiological
f. Bio Sino, Beijing, China parameters and cytokines of myelosuppressive mice.
g. SPSS 13.0 software, IBM Corporation, Armonk, NY, Chinese Veterinary Science 2013;43:537-544.
USA 15. Chinese ministry of health. The Evaluation Procedures
and Methods on Health Food Safety Toxicology [S]
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A Randomized and Controlled Experimental Study

on the Effects of Epimedium-Propolis Adjuvant on
Immune Function in Mice, Rabbits and Chickens
Yuanliang Hu DVM, PhD, Jiaguo Liu DVM, PhD, Dalu Song DVM, Zhigang Xu DVM
ABSTRACT
Four related randomized and controlled experiments were performed to evaluate the effects of Epimedium-Propolis adjuvant
(EPA) on T lymphocyte transformation rates in mice and chicks and antibody levels in rabbits and chickens. In Experiment
I, mice were divided into 4 groups and injected with the immunosuppressive agent cyclophosphamide (Cy), or EPA+Cy, or
EPA only and compared to untreated saline control. Results showed that Cy down-regulated T lymphocyte transformation
rate (SI), while EPA up-regulated it. In Experiment II, chicks were injected with either 0.2 ml or 0.4ml EPA and compared to
untreated saline control. EPA increased SI in a dose dependent and age related manner in chicks. In experiment III, infectious
canine hepatitis virus (ICHV) antibody level was evaluated in rabbits injected with virus + EPA and without EPA. Antibody
levels were higher in the EPA group. Experiment IV results demonstrated that EPA and Freund’s adjuvant (FA) created nearly
equal antibody increases in hens, however, EPA did not demonstrate the adverse side effects seen with FA. In conclusion,
EPA improved cellular and humoral immune function of animals in a dose dependent manner with the greatest intensity of
the effect in young animals. In addition, the use of EPA as an adjuvant is as effective as Freund’s adjuvant but without the
deleterious side effects on health. This study adds to the body of research demonstrating the benefit of using the Chinese herbal
medicine, Epimedium along with extracts from propolis as an adjuvant in vaccines for animals.
Key words: Epimedium-Propolis adjuvant, cyclophosphamide, T lymphocyte transformation rates, cellular immunity, hu-
moral immunity
ABBREVIATIONS chemicals are effective, they often have severe side effects
such as local inflammation, slow biodegradation and
EPA Epimedium-Propolis Adjuvant absorption, difficult preparation and inability to effectively
Cy Cyclophosphamide improve the immunogenicity of a weak antigen.2
PS Physiological Saline Epimedium-Propolis adjuvant (EPA) is s a combination of
FCS Fetal Calf Serum flavonoids from the Chinese herbal medicine, Epimedium,
FA Freund’s adjuvant combined with extracts from propolis. It has been shown
ICHV Infectious Canine Hepatitis Virus to be an effective immune enhancing agent in both
XOO Xanthomonas Oryzae Pv. oryzae clinical applications and experimental studies.3.4 In a
NK Natural Killer previous experimental study conducted by the authors,
PHA Phytohemagglutinin it was demonstrated that EPA enhanced the activity of
natural killer (NK) cells in peripheral blood, increased
SI Stimulation Index
the phagocytic activity of macrophages and increased
concentrations of cAMP and cGMP in plasma of chickens
An adjuvant is an immunological agent that can and mice.5,6
enhance the immune response to an antigen by an animal. The objective of this randomized, controlled study
They can be used as an immunostimulant to enhance was to add to this body of knowledge by determining in
vaccine response and can be injected prior to antigen 4 related experiments the effects of EPA on T lymphocyte
exposure or together with the antigen.1 Although widely transformation rates (SI) in 4 and 8 week-old mice and
used adjuvants such as oil emulsion, aluminum and some 3 day-old chicks when given EPA alone or with the
immunosuppressive agent cyclophosphamidea (Cy).
From: College of Veterinary Medicine, Nanjing Agricultural In addition, antibody levels were measured in viral and
University, Nanjing, 210095, China (Hu, Liu, Song, Xu)

bacterial challenged rabbits and hens, respectively, with determined by micro-whole blood culture 3H-TdR
and without administration of EPA. The hypothesis of (tritiated thymide) incorporation assay.3,7 The results
the study was that EPA would stimulate T lymphocyte are recorded as Stimulation index (SI). The formula to
transformation rates and perform as an efficetive adjuvant calculate this number was as follows: SI=PHA+ CPM
even in the face of an immunosuppressive agent and that (count per minute, mean of 3 tubes)/PHA- CPM (mean
it would have its greatest effect on young animals with an of 3 tubes). The same experiment was also conducted on
immature immune system. 8-week-old mice (Table 1).
In Experiment II, three hundred sixty healthy Roman
MATERIALS AND METHODS male chicksd (3-days-old) were randomly divided into 3
The experimental mice, chicks, hens and rabbits used groups. Chicks in Group 1 were injected with 0.2 ml PS,
in this study were housed, fed and watered conventionally. chicks in Group 2 were injected with 0.2 ml EPA, while
All care and experimental procedures were performed in chicks in Group 3 were given 0.4 ml EPA. Heart blood was
accordance with the guidelines of the Chinese Council for collected from each group at 7, 21, 35 and 49 days (Table
Animal Care. In Experiment 1, twenty-four healthy miceb 2). T lymphocyte transformation rate was determined as
(4-weeks-old) were randomly divided into 4 groups (n=6): described in Experiment I with the following exception: in
Group 1 was dosed with 25 mg/kg (Cy), Group 2 dosed Experiments I and II, the concentration of fetal calf serum
with 0.2 ml physiological saline (PS), Group 3 dosed with (FCS)e and phytohemagglutinin (PHA) had different
0.2 ml EPAc (flavones:20 mg/ml) + 25 mg/kg Cy, or Group results with CPM (RPMI 1640)f, so the concentration of
4 with 0.2 ml EPA. Mice were injected subcutaneously FCS and PHA that would stimulate CPM at the highest
once every 48 hours (hrs) in the lumbar muscles for a total level was selected as a culture and stimulation condition.
of three injections. Forty-eight hrs after the last injection, Therefore, in Experiment I, 10% FCS and 0.02% PHA
an orbital blood sample (heparin anticoagulation) was selected; while in Experiment II, 2.5% FCS and 10%
was collected. T lymphocyte transformation rate was PHA was selected.
Table 1: Experimental Groups and Test Material Dose; Experiment I

Group 1 2 3 4
Dose Cy (25 mg) PS (0.2mL) EPA (0.2mL)+Cy (25 mg/kg/BW) EPA (0.2 mL)
N= 6 6 6 6
Twenty-four healthy mice (4-weeks-old) were randomly divided into 4 groups. Group 1 was dosed with 25 mg/kg (Cy), Group
2 dosed with 0.2 ml physiological saline (PS), Group 3 dosed with 0.2 ml EPA + 25 mg/kg Cy, or Group 4 with 0.2 ml EPA.
Mice were injected subcutaneously once every 48 hours (hrs) in the lumbar muscles for a total of three injections. Forty-eight
hrs after the last injection, a blood sample (heparin anticoagulation) was collected. The experiment was repeated in 8-week-
old mice.
Table 2: Experimental Groups and Test Material Dose; Experiment II

Group I II III
Dose PS (0.2mL) EPA (0.2 mL) EPA (0.4 mL)
Blood Collected Days 7, 21, 35, 49 Days 7, 21, 35, 49 Days 7, 21, 35, 49
In Experiment II, three hundred sixty healthy Roman male chicks (3-days-old) were randomly divided into 3 groups. Chicks
in Group 1 were injected with 0.2 ml PS, chicks in Group 2 were injected with 0.2 ml EPA, while chicks in Group 3 were
given 0.4 ml EPA. Heart blood was collected (n=8) from each group at 7, 21, 35 and 49 days.
Table 3: Experimental Groups and Test Material Dose; Experiment III

Group A B
Dose EPA-ICHV (1 mL) ICHV (1 mL)
N= 4 4
Eight adult Chinchilla rabbits were divided into two groups: the experimental group with EPA adjuvant plus Infectious
Canine Hepatitis Virus (Group A) and the control group given ICHV without adjuvant (Group B). Rabbits in the 2 groups
were subcutaneously injected in the neck with 1 ml of EPA-ICHV or ICHV respectively, once per week repeated 4 times. Ear
vein blood was collected on the 7th day after the first vaccination and then repeated every 2 weeks for a total of 5 collections.

In Experiment III, eight adult Chinchilla rabbits were in the Freund’s Group. Xanthomonas oryzae pv. oryzae
divided into two groups: the experimental group with EPA (XOO) is a bacterium which is associated with rice blight.
adjuvant plus Infectious Canine Hepatitis Virus (Group A second injection was given to each group in the 10th week
A) and the control group given ICHV without adjuvant (when the antibody titer had decreased). In the FA group,
(Group B). Rabbits in the 2 groups were subcutaneously Freund’s complete adjuvant was given the first time, and
injected in the neck with 1 ml of EPA-ICHV or ICHVg Freund’s incomplete adjuvant (does not contain cell wall
respectively, once per week repeated 4 times (Table 3). of Mycobacterium tuberculosis to enhance immunization)
Ear vein blood was collected on the 7th day after the first was given at the second time. After the first vaccination,
vaccination and then repeated every 2 weeks for a total of 5 the effects of the two agents on the diet, life quality and
collections. Serum ICHV antibody OD490 was measured egg laying of hens were observed daily. The egg yolk
in each sample with indirect ELISA method.8 XOO antibody titer was determined by a double diffusion
In Experiment IV, six healthy Roman hens which had method, determined once daily at the start of egg laying
just started to lay eggs were divided into two groups: EPA by hens. Once the antibody was at measurable levels, the
group and Freund’s adjuvant (FA)h group (Table 4). Two detection was carried out every 3 days for three months.
ml of EPA + Xanthomonas oryxae pv. oryzae (XOO)I was Statistical significance for all 4 experiments in this
injected into the pectoralis muscle in the EPA group and 2 study was determined by using a T-test. This simple
ml of FA + XOO was injected into the pectoralis muscle statistical method was used since comparison was aimed
Table 4: Experimental Groups and Test Material Dose; Experiment IV

Group EPA-XOO FA-XOO
Dose EPA-XOO (2 mL) FA-XOO (2 mL)
N= 3 3
Six healthy Roman hens were divided into two groups: EPA group or Freund’s adjuvant (FA) group. Two ml of EPA +
Xanthomonas oryxae pv. oryzae (XOO) or 2 ml of FA + XOO was injected into the pectoralis muscle. A second injection was
given to each group in the 10th week (when the antibody titer had decreased).
Table 5: Effect of EPA on T lymphocyte transformation rate (SI) in mice; Experiment I

Group 4-Week-old 8-Week-old
1 (Cy) 1.08±0.04 1.14±0.96
2 (PS) 1.32±0.05** 1.30±0.19
3 (EPA+Cy) 1.24±0.08* 1.24±0.05
4 (EPA) 1.72±0.68** 1.40±0.10*
*P<0.05, **P<0.01; mice of same age compared to control
In 4-wk old mice, SI was greatest in Group 4 (EPA), while Group 1 (Cy) had the lowest value. The level of SI in Group 4 was
very significantly (P<0.01) higher than the other 3 groups, while the SI level in Group 1 was very significantly (P<0.01) lower
than the other 3 groups. These results indicate that EPA adjuvant could significantly enhance T lymphocyte transformation rate
while Cy could significantly inhibit it in 4-week-old mice. EPA adjuvant has an enhancing effect on the transformation of T
lymphocytes in 8-week-old mice and can antagonize the immunosuppression of Cy but the effect is weaker in the older mice
Table 6: Effect of EPA on T lymphocyte transformation rate (SI) in chicks; Experiment II

Group 7-day-old 21-day-old 35-day-old 49-day-old
1 (PS) 3.68±0.86 4.94±2.13 5.40±1.91 6.64±3.20
2 (EPA 0.2ml) 4.23±0.68 10.35±5.09** 9.20±4.28* 8.10±3.95
3 (EPA 0.4ml) 5.30±1.57 13.29±5.98** 11.34±3.34** 7.16±3.38
*P<0.05, **P<0.01; chicks of same age compared to control
Group 3 (0.4ml EPA) had the highest SI (Day 7, 21, 35), followed by Group 2 (0.2ml EPA), and Group 1 (saline) with the
lowest values. There was no significant difference among the 3 groups (P>0.05) at the age of 7 days and 49 days, while at
the age of 21 days and 35 days, SI in Groups 2 and 3 were significant or very significantly higher (P<0.05 or P<0.01) than
that in Group 1.

at only comparing the difference between the experimental no significant adverse effects on hens injected with EPA.
group and control group. Statistical significance was The hens had normal appetites by the 2nd day following
placed at P<0.05 (significant) or P<0.01 (very significant). injection and started egg laying 1 week later. The hens in
the FA group had severe adverse effects during the first 4
RESULTS days after injection demonstrated by loss of appetite, poor
The lymphocyte transformation rate (SI) in 4-week- health for 1 week and a delay of egg laying for 2 weeks.
old mice is shown in Table 5. SI was greatest in Group 4 After the second injection, the effects of the two different
(EPA), while Group 1 (Cy) had the lowest SI. The level adjuvants on health and egg laying of the hens were
of SI in Group 4 was very significantly (P<0.01) higher similar to the effects observed after the first injection (FA
than the other 3 groups, while the SI level in Group 1 compromised health of hens) with slightly less severity.
was very significantly (P<0.01) lower than the other 3 For both adjuvants, similar increasing antibody titers
groups. These results indicate that EPA adjuvant could were detected 1 week after inoculation with a high titer
significantly enhance T lymphocyte transformation rate level (1:64) still present 2 weeks post-inoculation. This
of 4-week-old mice, and Cy could significantly inhibit T high titer remained for at least two months with the second
lymphocyte transformation rate of 4-week-old mice. The injection boosting titer levels again to 1:256 at two weeks
level of SI in Group 3 (EPA+Cy) was also significantly post-inoculation (Figure 2).
(P<0.05) higher than Group 1, and its value was close to
that of Group 2 (saline). There was no significant difference DISCUSSION
between Groups 2 and 3 (P>0.05), which indicates that T lymphocytes are the primary cell involved in cellular
EPA adjuvant can effectively antagonize the immune immune function. There are several subgroups, which
suppressing effect of Cy on T lymphocyte transformation jointly constitute the immune regulatory network.9 The
rate of 4-week-old mice and return T lymphocyte activity T lymphocyte transformation rate reflects the functional
to normal level (Table 5). activation of T cells directly. Results from Experiment I
For 8-week-old mice, the SI values in each group showed that EPA adjuvant can significantly improve the
were consistent with those for the 4-week-old mice. transformation rate of T lymphocytes in mice at the age of
There was a difference between Groups 1 (Cy) and 4 4 weeks even when exposed to the deleterious effects of
(EPA) (P<0.05) but no significant difference among the an immunosuppressive agent such as cyclophosphamide.
other groups (P>0.05). This indicated that although EPA Experiment II demonstrated similar effects in that EPA
adjuvant has an enhancing effect on the transformation of adjuvant significantly improved the T lymphocyte
T lymphocytes in 8-week-old mice and can antagonize the transformation rate in chicks at ages 21 and 35 days. In
immunosuppression of Cy, it is a weaker effect in the older addition, Experiments I and II also demonstrated that the
mice (Table 5). intensity of the EPA adjuvant effect was both related to
Results of Experiment II showed that Group 3 chicks the dose and the age of the animals. The effect of the EPA
(0.4ml EPA) had the highest SI, followed by Group 2 adjuvant in 0.4ml/bird was greater than that in 0.2ml/bird,
(0.2ml EPA), and Group 1 (saline) with the lowest values. and its effect on 4-week-old mice and 21 and 35 day-old
There was no significant difference among the 3 groups chicks was more intense than on 8-week-old mice and
(P>0.05) at the age of 7 days and 49 days, while at the 49-day-old chicks (Table 5 and 6).
age of 21 days and 35 days, SI in Groups 2 and 3 were More mature animals have completed development
significant or very significantly higher (P<0.05 or P<0.01) of their immune organs while the immune organs and
than that in Group 1 (Table 6). immune function in young animals is still immature. This
Experiment III which evaluated the effect of EPA juvenile immune system allows the EPA adjuvant to have
on ICHV titers as depicted in Figure 1, show that OD490 a more significant positive effect on young animals as
values in rabbit serum of Group A (experimental group an immune enhancer alone or together with an antigen.
with adjuvant) were higher than those of Group B (control With enhanced function, young animals can improve their
group) without adjuvant. Significant (P<0.05) or a very early immune response ability and resistance to infectious
significant difference (P<0.01) was observed for all days disease. A further consideration is the use of EPA adjuvant
except the 7th day after the first vaccination. For Group B, in sick animals. It is expected that it would enable an
the antibody level increased slightly after vaccination, and animal that is suffering from a poorly functioning immune
reached its peak at the 21st day and then decreased rapidly, response the ability to more effectively confront infectious
with the 35th day containing antibody levels close to that agents.
of the 7th day. For Group A, the antibody level increased The use of EPA also significantly enhances the
significantly after vaccination and continued to rise at the humoral immune function. The results from Experiment
21st day. It peaked at the 28th day, then decreased slowly III demonstrated that the addition of EPA in a vaccine
through the 35th day, although, the antibody titer was still administered to study animals can significantly improve
significantly higher than at 7th day (P<0.01). the level of serum ICHV antibody production and can
In experiment IV, after the first injection, there were maintain a higher antibody titer for a longer time (Figure

Figure 1: Effect of EPA (red line) on serum ICHV antibody titers (OD490 Values) in rabbits; Experiment III
△ Inoculation date (0d, 7d, 14d, 21d)
Group A titers (EPA adjuvant group) were higher than those of Group B (control group). Significant (P<0.05) or a very
significant difference (P<0.01) was observed for all days except the 7th day after the first vaccination. For Group B, the anti-
body level increased slightly after vaccination, and reached its peak at the 21st day and then decreased rapidly, with the 35th
day containing antibody levels close to that of the 7th day. For Group A, the antibody level increased significantly after vac-
cination and continued to rise at the 21st day. It peaked at the 28th day, then decreased slowly through the 35th day, although,
the antibody titer was still significantly higher than at 7th day (P<0.01).
Titer
Figure 2: Effect of EPA (red line) and FA (blue line) on titers of yolk XOO antibody in hens; Experiment IV
△ Inoculation date (0d, 76d)
For both adjuvants, similar increasing antibody titers were detected 1 week after inoculation with a high titer level (1:64)
still present 2 weeks post-inoculation. This high titer remained for at least two months with the second injection boosting
titer levels again to 1:256 at two weeks post-inoculation. Results showed that EPA stimulated antibody production levels
similar to those of Freund’s adjuvant, without the adverse side effects.
1). In Experiment IV, the EPA adjuvant and Freund’s REFERENCES

adjuvant had similar egg yolk XOO antibody titers. 1. Zhang ZX, Jiang P, editor in chief. Practical
Differences between the 2 adjuvants, however, occurred Veterinary Biological Products Technology. Chinese
when technical production difficulty of the adjuvant and Agricultural Science and Technology Press, 1996:
adverse health effects were examined. There is a greater 50-55.
delay in egg laying with FA along with greater adverse 2. Xie QG. Research Progress on Immune Control of
effects on life quality. In addition, the EPA adjuvant Major Epidemic Diseases of Livestock and Poultry.
is technically simpler and faster to produce. It is an Chinese Agricultural Science and Technology Press,
aqueous solution, which is more readily mixed with the 1995:143-168.
antigen and does not have a long emulsion time like FA 3. Gui ZY, Geng PL. The Regulation Effect and
(0.5-1 hrs) which is associated with greater loss of the Mechanism of the Herbal Epimedii Mixture and its
antigen. Finally Freund’s adjuvant vaccine has a higher Extract on the Immune Activity of the Experimental
viscosity compared to EPA, making it difficult to use for Animal. Chinese Journal of Immunology, 1985, 1
subcutaneous skin injections. (1): 25-29.
In conclusion, the EPA adjuvant improves both 4. Cheng QL, Chen XM. Changes of Immune Function
cellular and humoral immune functions. The hypothesis of in Rats with Renal Resection and the Regulation of
the study was proven through demonstration of enhanced Chinese Herbal Medicine Epimedii. Chinese Journal
immune function through increasing T lymphocyte of Microbiology and Immunology, 1993, (3): 198-
transformation rate even when animals were exposed to 200.
cyclophosphamide. In addition, the EPA adjuvant was 5. Liu JG, Hu YL, Zhang BK et al. Effect of EPAAdjuvant
associated with prolonged increases of serum antibody on the Activity of NKC and Peritoneal Macrophages
levels in rabbits exposed to an infectious agent and also in Mice. Chinese Journal of immunology, 1996, 12
stimulated antibody production levels similar to those of (Suppl1): 240-242.
Freund’s adjuvant, without the adverse side effects. The 6. Liu JG, MING K, Hu YL. Effect of Epimedium-
experiments conducted in this study add to the body of Propolis Adjuvant on the Nonspecific Immunologic
research demonstrating the benefit of using Epimedium- Functions of Experimental Animals. American
Propolis as an adjuvant in vaccines for animals. Journal of Traditional Chinese Veterinary Medicine,
2016, 11(1): 29-33.
Footnotes 7. Yang GZ, editor in chief. Medical Immunology, Jilin
a. Shanghai Hualian pharmaceutical company People’s Publishing House, 1981:381-385.
b. Nanjing General Hospital of Nanjing Military 8. Department of Microbiology, Beijing Medical
Command Experimental Animal Center College, Experimental immunology. People’s
c. Institute of Traditional Chinese Veterinary Medicine, Medical Publishing House, 1980: 273-283.
College of Veterinary Medicine, Nanjing Agricultural 9. Wu JG. Practical Clinical Immunology Test. Jiangsu
University. It contains 1% Epimedium total flavone Science and Technology Press, 1990, pp 3-7.
and 10% Propolis
d. A state-owned chick farm
e. Gibco products, United States
f. Culture media developed by Roswell Park Memorial
Institute; Gibco products, United States
g. Provided by the Nanjing Police Dog Institute
h. Gibco products, United States, including the complete
adjuvant and incomplete adjuvant
i. Provided by the Department of Plant Pathology

Clinical Studies
A Randomized Controlled Study Comparing Da Xiang

Lian Wan to Metronidazole in the Treatment of Stress
Colitis in Shelter/Rescued Dogs
Margaret P Fowler DVM, MS, Deng-Shan Shiau PhD, Huisheng Xie DVM, PhD
ABSTRACT
Traditional Chinese Veterinary Medicine (TCVM), which can include treatment with Chinese herbal medicine, is being used
increasingly in Western medicine to treat a variety of diseases in dogs. With the increased interest in Chinese herbal medicines,
evidence-based research to prove efficacy and safety of these traditional medicines is necessary. The objective of this research
was to compare the effectiveness of the Chinese herbal medicine, Da Xiang Lian Wan (DXLW), to the Western pharmaceutical,
metronidazole, in the treatment of stress colitis in sheltered/rescued dogs utilizing a randomized controlled study with a
hypothesis that successful treatment with DXLW would not be less than metronidazole. Fifty-six dogs were randomly assigned
to either metronidazole treatment group or DXLW treatment group. Each medication was administered orally twice daily at
recommended clinical doses for a maximum of 10 doses. The results indicate that dogs in the metronidazole treatment group
had an 89% response rate (normal stools) within the 10 dose protocol while the DXLW group had a 97% positive response rate
(p < 0.05, non-inferiority test). This study demonstrates that DXLW is as effective as metronidazole in resolving stress colitis
in sheltered dogs and can be an effective alternative treatment for patients that do not respond to metronidazole or cannot
tolerate it.
Key words: colitis, dog, stress, traditional Chinese veterinary medicine, Da Xiang Lian Wan, Coptis, Saussurea, Huang Lian,
Mu Xiang, Great Saussurea Coptis
ABBREVIATIONS from fear, anxiety and sometimes over-crowding. Their

gastrointestinal tract is also physiologically stressed from
diet changes. While no medical literature could be found
discussing the role of stress in dogs with colitis, in humans
DXLW Da Xiang Lian Wan
a higher perceived level of stress in patients appeared to
BID twice daily
be a contributing factor in ulcerative colitis and Crohn’s
TID three times daily
disease and experimental work with mice demonstrated
that stress aggravated existing colitis. 3,4
Stress colitis is a common problem in rescued or
Colitis and ulcerative colitis in dogs and other species
sheltered dogs housed in a kennel type environment and in
have been well discussed in veterinary journals. Several
the case of kill shelters, can result in euthanasia of affected
conventional medications have been used as a treatment
dogs.1 Colitis is defined as a high frequency of defecation
addition or alternative to metronidazole, including
with small fecal volume. Affected dogs often display
tylosin, fluoroquinolones, azathioprine, sulfasalazine,
prolonged tenesmus after defecation and the loose stools
amoxicillin and prednisone. Often no one single agent
typically contain mucus and/or blood.2 The syndrome may
such as metronidazole is effective and combinations
be multi-factorial involving gastrointestinal parasites,
of medications can be required. 5 The role of bacteria
bacterial or viral infections, diet changes, anxiety and
in the development of colitis has been discussed and
stress. Abandoned dogs in kennel type facilities are
demonstrated along with the failure of antibiotics such
particularly under a high degree of psychological stress
as metronidazole in the treatment of canine colitis due to
antibiotic resistance. 5,6,7
From: Acupuncture & Holistic Veterinary Services of Northwest
Florida, Panama City Beach, Florida (Fowler); Chi Institute, Metronidazole is listed as the drug of choice for
Reddick, Florida, and Optima Neuroscience, Inc., Alachua, Florida Inflammatory Bowel Disease and Large Bowel Diarrhea
(Shiau); Chi Institute, Reddick, Florida (Xie).

(colitis) at dosages in dogs of 10-15 mg/kg of body weight and Police Dogs. A well designed experimental study
two (BID) to three (TID) times daily or 20-25 mg/kg could also provide further scientific evidence to validate
BID.8 It is typically effective, inexpensive, and is the drug the use of Chinese herbal medicine in Western clinical
of choice for colitis in shelter facilities at dosages of 10- practice. The aim of the study was to provide evidence
22 mg/kg of body weight BID. Metronidazole, however, based experimental support of an effective alternative
carries a risk of adverse reactions such as neurotoxicity treatment for stress diarrhea in dogs. The objective of this
including ataxia and seizures, lethargy, weakness, randomized, controlled study focused specifically on stress
neutropenia, hepatotoxicity, hematuria, anorexia, nausea, colitis in shelter dogs and compared treatment efficacy of
vomiting and diarrhea, and may be teratogenic.8,9 These Da Xiang Lian Wan to metronidazole. The hypothesis was
toxic side-effects, in addition to poor clinical response in that Da Xiang Lian Wan was at least as effective (speed of
some dogs, limit the drug’s usage in a subset of shelter diarrhea resolution, % response rate of treated dogs) as the
patients. conventional medication metronidazole in the treatment
With metronidazole’s ineffectiveness and intolerable of stress colitis in dogs held in a rescue dog sanctuary or
side effects for some patients, alternative treatments are shelter.
being considered by pet owners, shelters and veterinarians
in conventional clinical practice. In traditional Chinese MATERIALS AND METHODS
veterinary medicine, disease patterns (TCVM patterns) Over a period of 8 months, there were a total of
are identified and associated with imbalances. Colitis, as 56 dogs enrolled in the study located at one of the 3
defined in this study, is consistent with a TCVM Excess participating animal shelters in the United States: Alaqua
Pattern of Large Intestine Damp Heat.10 Chinese herbal Animal Refuge located at 914 Whitfield Road, Freeport
medicines are formulated to correct these imbalances FL, 32439, Low Country Lab Rescue located at 1612
to assist healing and restore health. Da Xiang Lian Wan Harbor View Rd, Charleston, SC 29412, and Cape May
(Great Saussurea Coptis®) a is an ancient Chinese herbal County Animal Shelter located at 110 Shelter Road, Cape
medicine to clear Damp-Heat, move Qi, relieve pain and May Court House, NJ 08210. Shelters were screened to be
is appropriate for the Excess Pattern diagnosed in enrolled certain a staff veterinarian supportive of the research was
study dogs (Table 1).10 available and that shelter staff members were adequately
In contrast to metronidazole, DXLW lists no trained on the importance of adhering to study protocols,
contraindications for its use.10 A review of scientific and testing procedures and medication administration.
medical literature was conducted on the use of DXLW When a shelter dog presented with acute onset
and its ingredients in dogs prior to this study, and it was diarrhea, a standardized flowsheet was utilized to
determined that very little research information on this determine eligibility for the study (Figure 1). Inclusion
subject was available. In relation to the herbal formula Da criteria for the study from this flowsheet required that
Xiang Lian Wan and the individual ingredient herbs, Huang the dog must be older than 5 months of age and have an
Lian (Coptis) and Mu Xiang (Saussurea), some references odiferous, bloody or mucus diarrhea typical of a colitis
could be found on scientific research conducted in-vitro or or frequent small volumes of loose stool with straining.
in-vivo in humans or non-canine species.11,12,13,14,15 There If a dog qualified through the first 3 steps of the flow
was, however, no statistically valid controlled randomized sheet (Figure 1), a fecal sample was then obtained and a
research studies using dogs as subjects. There were several fecal floatation and an IDEXX Giardia Snap Test b were
claims of the effectiveness of this Chinese herbal medicine run to rule out contributing parasites. Flowsheets on
for the treatment of colitis, diarrhea or inflammatory bowel dogs that did not qualify based on parasite test results
disease in dogs, but none had been extensively studied in were retained for further review (19 dogs). Exclusion
dogs. 10,12,16,17,18 criteria included subjects with a positive fecal flotation
With the potential clinical benefit of less side effects parasite test or IDEXX Giardia Snap Test, watery diarrhea
than metronidazole, successful efficacy validation of typical of a TCVM Deficiency pattern diarrhea (i.e.
DXLW would support its use in rescue dogs with stress Spleen Qi Deficiency) or evidence of severe debilitation,
related colitis, the general pet dog population with emaciation, or other serious known health issues (i.e.
colitis, as well as stress prone Military Working Dogs kidney insufficiency, heart failure, liver disease, life-
Table 1: Ingredients and actions of Da Xiang Lian Wan (Great Saussurea Coptis®)
English Name Chinese Pin-Yin Ingredient % Action
Saussurea Mu Xiang 50 Move Qi and relieve pain
Coptis Huang Lian 25 Clear Damp-Heat in the intestines
Evodia Wu Zhu Yu 25 Warm the Middle Jiao and stop vomiting
Manufactured by Jing Tang Herbal, Reddick, FL, USA

Enroll a patient in Herbal vs. Metronidazole Study
No
Exclude from
Is the patient at least 5 months
the study
old?
Yes
Is the patient critically

No Exclude from
ill, emaciated, having cancer, known
liver or kidney failure, or watery the study
diarrhea?
Yes
Does the patient have

diarrhea with any of the following? No Exclude from
(1) mucus, (2) fresh blood in stools,
the study
(3) frequent small volume stinky
stools, (4) straining as
in colitis?
Yes
No
Is the patient negative on Exclude from
Fecal Float AND Giardia the study
Snap Test?
Yes
Include in the study
Figure 1: Subject recruitment form used at all participating shelters for study dog enrollment.
threatening cancer). Dogs that qualified for the study were 10 doses and administration of each dose was recorded.
randomly assigned to either the metronidazole treatment All medications were administered directly by trained
group or the DXLW treatment group using a commercial personnel preferably with or immediately after feeding.
software program c. For each participating facility, a A chart was supplied to each shelter by the investigator
separate list of computer generated random assignment (Fowler) with preset DXLW doses which were calculated
numbers based on the anticipated number of eligible based on weight ranges that fell within the recommended
dogs was utilized to prevent assignment bias. The study dose (Table 2). The same approach was predominately
identification number, the facility identification, age, sex, used for dosing metronidazole. This was done to provide
dose, weight, and breed were recorded for each dog. Each consistency and simplify study dosing calculations for
facility had its own data sheet to record information on shelter staff irrespective of who was attending the study
each dog in the study (Figure 2). The study was conducted dog on a particular day. Based on the weight ranges,
unblinded in order to efficiently use the small pool of metronidazole was orally dosed at 10-22 mg per kilogram
trained personnel available to conduct this study in a of body weight twice daily and DXLW was orally dosed at
shelter environment. Each medication was administered 0.5 grams per 10-20 pounds of body weight twice daily.
in the morning and late afternoon, for a maximum of Trained technicians/caregivers documented the

Table 2: Da Xiang Lian Wan Dosing Protocol*

< 5# 5.1- 10.1- 15.1- 19.1- 25.1- 30.1- 45.1- 60.1- >80#
BW 10# 15# 19# 25# 30# 45# 60# 80# BW
BW BW BW BW BW BW BW BW
0.2g capsule 1 2
0.5g capsule 1 1.5 2 2.5 3 4 5 6
BW= Study Dog’s Body Weight *Each study dog was dosed twice daily based on a weight range
ALAQUA DATA SHEET FOR HERBAL/METRONIDAZOLE STUDY

DATE:
1. Has Dog Qualified from Flowsheet? If Yes, attach flowsheet and continue. Keep Excluded flowsheets and note reason
for Exclusion.
2. Randomly assign to Herbal or Metronidazole group based on admission sequence (below).
HERB METRONIDAZOLE
HERB GROUP: Consists of Dogs 1 3 6 9 10 11 12 15 20 22 23 25 27 28 30
METRONIDAZOLE GROUP: Consists of Dogs 2 4 5 7 8 13 14 16 17 18 19 21 24 26 29
3. Dose prescribed:
4. Dog ID: Study Number ( ) Alaqua ID ( ) AGE: SEX: WEIGHT: BREED:
COMMENTS (i.e. vomiting, not eating, did

DAY DIARRHEA MEDS GIVEN
not take meds, died, etc.)
AM PM AM PM
1 Y N Y N Y N Y N
2 Y N Y N Y N Y N
3 Y N Y N Y N Y N
4 Y N Y N Y N Y N
5 Y N Y N Y N Y N
HERBAL: Give for 2 doses AFTER diarrhea has resolved up to a MAXIMUM of 10 doses. Give preferably with food in
stomach.
Metronidazole: Give for 10 doses.
Figure 2: Sample Data Collection Form used by all participating shelter/rescue facilities to record study information for each
dog each day.

presence or absence of diarrhea twice daily (in the proportions (first non-inferiority hypothesis). The null
morning and late afternoon), recorded the administration hypothesis would be rejected if the lower bound of the
of DXLW or metronidazole, and recorded the number of 95% confidence interval was greater than the preset non-
doses of DXLW or metronidazole required to resolve the inferiority limit (-0.15). A 95% confidence interval of the
diarrhea. DXLW treatment was continued for 2 doses after mean dose difference (DXLW Group – Metronidazole
resolution of diarrhea or 10 doses, whichever came first. Group) was calculated based on the two-sample t-test for
Metronidazole treatment was administered for a total of equality of means (second non-inferiority hypothesis).
10 doses. Minor adverse reactions including vomiting The null hypothesis would be rejected if the upper
and any major adverse reactions including mortality were bound of the 95% confidence interval was less than the
documented. Diarrhea was considered resolved when the preset non-inferiority limit (2 doses). It was anticipated
clinical signs of colitis including blood or mucus in the that the standard metronidazole treatment would have a
stools or tenesmus were absent. Study dogs had to then success rate of 75% and the DXLW treatment a success
remain free of clinical signs of colitis for a 24-hour period rate of 90%. With the non-inferiority limit set to be 15%,
before qualifying as a treatment success. Subjects were 54 subjects (27 per group) were required to reach a test
considered a treatment failure if clinical resolution failed power of 90% to reject the null hypothesis (inferiority)
to occur during the treatment protocol period or within 24 with 95% confidence. With this sample size, the study
hours of completing the treatment protocol. could guarantee a 90% power to reject the null hypothesis
Two outcome measures were recorded for statistical (inferiority) with 95% confidence, with the non-inferiority
analysis; either the diarrhea resolved or persisted within limit set to be 2 doses. For each of the statistical tests, a
10 doses, and the number of doses that were required for p-value < 0.05 was considered statistically significant.
resolution of clinical signs was tracked as continuous
measures data. The two statistical hypotheses were: RESULTS
(1) Within 10 doses of the treatment, the proportion of All 56 dogs which were enrolled completed this study
subjects successfully treated by the proposed DXLW with 29 in the DXLW Group and the remaining 27 in the
treatment is not smaller than (i.e., not inferior to) that by Metronidazole Group. Study dogs were randomly assigned
the standard metronidazole treatment and (2) For subjects to study groups, therefore, a comparison of the distribution
successfully treated by either the DXLW or metronidazole of gender, age, and body weight of study animals between
treatment, the mean number of dosages required for the the 2 groups was evaluated for effects on study results. Of
DXLW treatment is not more than that required for the the 56 dogs in the study, 35 (62.5%) were male or neutered
metronidazole treatment. males and 21 (37.5%) were female or spayed females.
A 95% confidence interval of the success rate In the Metronidazole Group, 18 were male and 9 were
difference (DXLW Group – Metronidazole Group) was female, a 2:1 ratio. In the DXLW Group, 17 were male and
calculated based on the two-sample test for equality of 12 were female, an approximately 3:2 ratio (Figure 3). The
Figure 3: Gender distribution of study dogs with an overall population of 35 males and 21 females. The Metronidazole
Group contained 18 males and 9 females (2:1 ratio) while the DXLW Group contained 17 males and 12 females (3:2 ratio).
The difference in gender between the 2 groups was not statistically significant (p = 0.73).
mean age of the entire study subject population was 4.30 DXLW Group had a higher proportion of females relative
years (range 0.4 - 13.0), with the Metronidazole Group to the entire population and the Metronidazole Group, but
mean at 3.64 years (range 0.4 – 10.0) and the DXLW the difference was not statistically significant (p = 0.73).
Group mean at 4.91 years (range 0.9 – 13.0) (Figure 4). Since there is no evidence that a certain gender would
The mean body weight over all study subjects was 38.2 respond better to either treatment, this difference was
pounds (range 4.4 – 84.0) with the Metronidazole Group considered not to have impacted the results of the study.
averaging 42.0 pounds (range 4.4 – 75.0) and the DXLW Age and body weight distribution between the groups were
Group 34.6 pounds (range 6.8 – 84.0) (Figure 5). The evaluated with a two-sample t-test. Again, the mean age
Figure 4: Mean age of the overall population of study dogs is 4.30 (range 0.4- 13.0) years. The Metronidazole Group’s mean
age is 3.64 (range 0.4- 10.0) years and the DXLW Group has a mean of 4.91 (range 0.9- 13.0) years. There was no statistically
significant difference in mean age between treatment groups (p =0.169).
Figure 5: The body weight distribution of study dogs with the overall population mean weight equal to 38.2 pounds (range
4.4-84.0). The Metronidazole Group had a mean of 42.0 (range 4.4-75.0) and DXLW Group a mean of 34.6 pounds (range
6.8-84.0). There was no statistically significant difference in mean weight between treatment groups (p =0.284).

and body weight comparison between the two treatment within 10 doses or less (Figure 6). The mean dose for
groups was not significantly different (p = 0.169 and p = successful resolution of the stress induced colitis was
0.248, respectively) which reduced the possibility that age 3.46 doses for metronidazole and 3.26 doses for DXLW.
or body weight would affect the outcomes of the study. There was no mortality associated with the administration
Of the 27 dogs entered into the Metronidazole of either study therapeutic compound (Figure 7). Adverse
Group, 24 (88.9%) had resolution of their colitis type reactions were minimal and limited to vomiting in 1 dog
diarrhea within the 10 dose protocol while out of the 29 in the Metronidazole Group which resolved with anti-
dogs entered in the DXLW Group, 28 (96.6%) resolved emetics and 1 dog in the DXLW Group which resolved
Figure 6: Proportion of successful treatment for resolving the colitis type diarrhea in study dogs with either metronidazole
(88.9%) or DXLW (96.6%) within 10 doses, p < 0.05 in testing the non-inferiority of DXLW group against Metronidazole
Group.
Figure 7: Box plot: Comparison of the observed mean dose required to resolve the colitis type diarrhea in the metronidazole
(3.46 doses) and DXLW (3.26 doses) treatment groups (p < 0.05, non-inferiority testing). The two treatments had very simi-
lar distribution of the “effective” doses, as both the median (the bold horizontal black line inside the box) and the range (the
height of the box and between the two whiskers) are very close between the two distributions. The only obvious difference
is an outlier in the DXLW group (the data point above the upper whisker). This particular subject required 9 doses to resolve
the colitis type diarrhea.
without treatment. The 4 dogs that failed to resolve with dogs.

treatment were equally represented by male and female, Although no clinical research studies on colitis or
with an age range of 2 to 10 years, and a weight range stress induced colitis in dogs were found, several TCVM
of 33 to 62 pounds. There was no commonality in their textbooks describe the effectiveness of DXLW herbal
representation. formula or the individual herbs contained within it for
To test the hypothesis that, within 10 doses of the the treatment of gastrointestinal disorders including those
treatment, the proportion of subjects successfully treated conventionally labeled as colitis. 10,16,17 In both TCVM
by the proposed DXLW treatment is not smaller than (i.e., clinical and herbal textbooks, DXLW is recommended
not inferior to) the standard metronidazole treatment, the for treatment of the TCVM pattern Damp Heat or Heat
study applied both the two-sample proportion test and the Toxin at a dosage of 0.5 grams per 10-20 pounds body
bootstrap method to generate the 95% confidence interval weight. Its excellent effects when used as a treatment for
of the proportion difference between the two groups, as the Western diagnosis of Inflammatory Bowel disease
described in the previous section. With the observed is described.10,16 The single herbs, Huang Lian (Coptis)
successful treatment rate at 0.89 (24/27) and 0.966 and Mu Xiang (Saussurea) are cited in Xie’s Chinese
(28/29) for the Metronidazole Group and DXLW Group, Veterinary Herbology text for Damp Heat in the intestines
respectively, since the 95% confidence interval of the with clinical signs of “Bloody mucous diarrhea, dysentery,
proportion difference calculated based on the two-sample and tenesmus”.17 Similarly, Mu Xiang combined with
test for equality of proportions has a lower bound greater Huang Lian is listed as an excellent Chinese herbal
than the preset non-inferiority limit -0.15 and even more medicine for Damp Heat dysentery.17 Other journal
significant result obtained through the bootstrap method, articles recommended further research on plants which
the null hypothesis could be rejected (p < 0.05). have been recommended for gastrointestinal disorders
Similar statistical evaluation of the hypothesis that, including Inflammatory Bowel Disease.19,20
for subjects successfully treated by either the DXLW or In this research project, through the design of a
metronidazole treatment, the mean number of doses randomized controlled clinical study, the investigators
required for the DXLW treatment is not more than that sought to demonstrate statistical evidence to support
required for the metronidazole treatment, the study applied the hypothesis that treating stress colitis in sheltered
both the two-sample t-test and again the bootstrap method dogs with the Chinese herbal DXLW is just as effective
to generate the 95% confidence interval of the difference as treating them with the conventional treatment of
of means between the two treatment groups. With the metronidazole. Dogs from 3 shelters were enrolled in the
mean dose required for success at 3.46 and 3.25 doses for study. An enrollment flow sheet which applied inclusion
the Metronidazole Group and DXLW Group, respectively, and exclusion criteria to each potential study dog was
since the 95% confidence interval of the mean dose performed. Overall distribution differences in gender,
difference calculated based on the two-sample t test for body weight and age were evaluated and found to be
equality of means had an upper bound smaller than the statistically insignificant between the 2 treatment groups,
non-inferiority limit of 2 doses and similar results obtained which suggests that these subject characteristics were
through the bootstrap method, the null hypothesis could be deemed unlikely to impact the outcomes of the study.
rejected (p < 0.05). The most common reason for exclusion from the
study for dogs that had qualified in all other enrollment
DISCUSSION aspects (19 potential subjects) was disqualification due to
Pet owners are increasingly seeking out herbal positive parasite tests. This represented 25% of otherwise
therapies to replace conventional pharmaceuticals which eligible presenting subjects, a higher than anticipated
often have adverse side-effects. Animal shelters are also ratio. IDEXX Giardia Snap Tests were positive in 84%
frequently looking at TCVM as a better way to manage of the parasitized dogs demonstrating that Giardia was
their populations, prevent and treat medical issues. The the primary disqualifying parasite, but additionally 21%
test formula selected to evaluate in this experimental study, of the potential subjects were positive for Ancylostoma
DXLW (Table 1) was identified as a good Chinese herbal (hookworms), 11% for Isospora (coccidia), and 5% for
formula to use because of the safety and efficacy observed Trichuris (whipworms). This higher than anticipated ratio
when treating canine colitis in one of the investigators’ by both the investigator (Fowler) and attending shelter
TCVM practice (Fowler), even in cases in which other veterinarians unfortunately encumbered the study through
conventional treatments such as metronidazole had failed. subject disqualification. It also resulted in one shelter
Additionally, a need to identify a safe, cost-effective changing their protocol to include an IDEXX Giardia
alternative medication to the limited choices available to Snap Test.
treat stress related colitis in sheltered dogs was needed. Based on the 56 dogs that qualified and completed the
The formula’s effectiveness in other species, including study, 89% of the animals assigned to the Metronidazole
humans, is well demonstrated in medical literature which Group responded to the treatment, while 97% of the
also made it attractive to assess as a treatment strategy for DXLW Group had successful resolution of their stress

colitis. These observations provided statistical evidence species. The 97% success rate of DXLW in treating
(p < 0.05) that the success rate of DXLW Chinese herbal sheltered dogs with stress colitis demonstrated in this study
medicine is not inferior to that of the standard treatment is close to the 90% success rate in humans cited in the
Metronidazole when treating stress colitis in sheltered literature review. 12 This research helps to bridge the gap
dogs. In addition, study dogs that responded to the assigned that exists between human and non-canine species versus
treatment (52 out of 56), had a mean dose of 3.46 doses dogs on the subject of DXLW and provides the scientific
in the Metronidazole Group and 3.26 doses in the DXLW community with validation of the efficacy of DXLW
Group (3.26 versus 3.46; p < 0.05). These observations in stress colitis in sheltered dogs. This validation also
statistically support that DXLW works as quickly as the supports the use of DXLW in stress colitis prone Military
current conventional treatment metronidazole. Working, Search and Rescue, and Police Dogs. The pet dog
Based on the population in the investigator’s TCVM population prone to colitis benefits as well by providing an
practice (Fowler), where most colitis affected dogs were effective alternative treatment to metronidazole and other
failed or untreatable with metronidazole, the anticipated conventional colitis medications, with fewer potential side
success rate of metronidazole was postulated to be 75%. effects. Additionally, this research helps to validate the
The measured success rate for study dogs, however, effectiveness of Chinese herbal medicine in general.
was 89%. The higher than anticipated success rate of In conclusion, DXLW, is an effective alternative to the
metronidazole in this study is most likely due to the fact standard metronidazole treatment in treating stress colitis
that the study subjects represented acute cases or acute in sheltered dogs. It is easily administered, is free of any
exacerbations of stress induced colitis. This is in contrast significant adverse side-effects and is not encumbered
to the often more chronic cases of colitis and ulcerative by antibiotic resistance seen in some cases. Lastly, this
colitis seen in the general dog population and veterinary research project lays the groundwork for a more detailed
practices. With multiple treatments of metronidazole and study to further validate DXLW and other Chinese herbal
other antimicrobials, presumably antibiotic resistance medicines. A well-funded study at a major university
becomes a factor in metronidazole failure in dogs seen in could recruit a larger number of eligible subjects and test
conventional practices.18,20 additional measurable indices such as routine clinical
While the anticipated success rate of DXLW was pathology, inflammatory factors, hormones, etc. Potential
90%, the final study results demonstrated a 97% success subjects could also be evaluated by an experienced TCVM
rate which again was better than the anticipated success veterinarian using tongue and pulse observations and
rate. This may be associated with DXLW not being other TCVM diagnostic criteria. These potential subjects
encumbered by microbial antibiotic resistance as a cause could then be diagnosed with a specific TCVM Pattern
of treatment failure in canine colitis. Treatment failures to determine that an Excess Pattern of Intestinal Damp
associated with DXLW administration are typically the Heat appropriate for DXLW qualified them for the study.
result of inappropriate use due to incorrectly identified Possible Deficiency patterns for which DXLW would not
TCVM patterns. After study completion, some dogs in be appropriate would then more effectively be excluded
both treatment groups, that had met treatment success from the study. A more detailed medical and background
criteria during the study, had stress colitis relapses as long history, which is not available in rescued and abandoned
as they continued to be held in the facility. In these cases, dogs, could be taken and yield additional useful
following recovery, administration of a Chinese herbal information. Additionally, DXLW could be compared to
medicine such as Xiang Lian San could be considered for other conventional medications used to treat colitis in
effective long term maintenance. 10 dogs and evaluated for bacterial resistance, the postulated
Useful information provided by the shelter reports factor in failure of conventional antimicrobial colitis
indicated that encapsulated DXLW was as easily medications. This could offer insight into DXLW efficacy
administered in food as metronidazole. Purchased in in resolving cases that have failed with metronidazole
bulk powder, the formula DXLW was cost effective even treatment.
for animal shelters at less than 10 cents per gram. The
investigator (Fowler) observed that the higher end DXLW ACKNOWLEDGMENTS
dose of 0.5 grams per 10 pounds body weight worked This work was financially supported by donations
more effectively and quickly at resolving the stress colitis from IDEXX Laboratories, The Chi Institute of Traditional
than the lower end of 0.5 grams per 20 pounds body Chinese Veterinary Medicine, Janet Stoner, Alicia Crew,
weight. In future studies, administering DXLW for the full Lynne Watling, and many of the clients of Acupuncture
10 doses rather than discontinuing treatment 2 doses after & Holistic Veterinary Services of Northwest Florida. The
the cessation of diarrhea may be even more effective, and authors would like to thank the veterinarians and staff
help prevent relapses. of Low Country Lab Rescue, Cape May County Animal
This research is in agreement with previously Shelter, Alaqua Animal Refuge and specifically Dr. Amy
conducted studies on the efficacy of DXLW and its Williams, for the donation of time and effort in the conduct
ingredients in treating colitis in humans and non-canine of this study.

FOOTNOTES 10. Xie H. Chinese Veterinary Herbal Handbook

a. Great Saussurea Coptis, Jing Tang Herbal, Reddick, 2nd Edition. Reddick, FL: Jing Tang Publishing
Florida, USA 2008:14,15, 247, 248.
b. IDEXX Laboratories, Inc., Westbrook, Maine 11. Cho JY, Baik KU, Yoo ES, et al. In Vitro
c. “R” version 3.2.2; The R Foundation for Statistical Antiinflammatory Effects of Neolignan Woorenosides
Computing, Vienna, Austria from the Rhizomes of Coptis japonica. J. Nat. Prod.
2000; 63(9):1205–1209.
REFERENCES 12. Zhang C, Ma A, Xie H. Review of Clinical and
1. Fowler, M P. Personal Communication: attending Experimental Research from China on 8 Chinese
veterinarians of Alaqua Animal Refuge, Cape May Herbal Medicines for Gastrointestinal Disorders.
County Animal Shelter, Walton County Animal American Journal of Traditional Chinese Veterinary
Shelter, Low Country Lab Rescue, and organizers of Medicine 2014; 9(2):53-68.
Florida Greyhound Rescue. Dec 2015-March 2016. 13. Fan H, Liu XX, Zhang LJ, et al. Intervention effects of
2. Tilley LP, Smith FWK. Blackwell’s Five Minute QRZSLXF, a Chinese medicinal herb recipe, on the
Veterinary Consult: Canine and Feline (5th Edition). DOR-β-arrestin1-Bcl2 signal transduction pathway
Ames, Iowa: Wiley Blackwell 2011:274-275. in a rat model of ulcerative colitis. J Ethnopharmacol
3. Tabibian A, Tabibian JH, Beckman LJ, et al. 2014; 154(1):88-97.
Predictors of health-related quality of life and 14. Luo Y, Zhao H, Liu Z, et al. Comparison of the
adherence in Crohn’s disease and ulcerative colitis: enteric mucosal immunomodulatory activity of
implications for clinical management. Dig Dis Sci. combinations of Coptis chinensis Franch. Rhizomes
2015; 60(5):1366-1374. and Evodia rutaecarpa (Juss.) Benth. Fruits in mice
4. Schultz M, Lai C, Lindstroem A, et al. Aggravation of with dextran sulphate sodium-induced ulcerative
Established Colitis in Specific Pathogen-free IL-10 colitis. Planta Med 2010; 76(8):766-772.
Knockout Mice by Restraint Stress Is Not Mediated 15. Yu LZ, Wu JY, Luo JB, et al. Effects of different
by Increased Colonic Permeability. J Crohns Colitis compositions of Gegenqinlian decoction on
2015; 9(9):754-762. experimental shigellosis in rabbit. [Article in
5. Hostutler RA, Luria BJ, Johnson SE, et al. Antibiotic- Chinese]. Di Yi Jun Yi Da Xue Xue Bao 2005;
responsive histiocytic ulcerative colitis in 9 dogs. J 25(9):1132-1134.
Vet Intern Med 2004; 18(4):499-504. 16. Xie H, Wedemeyer L, Chrisman C, et al. Practical
6. Menozzi A, Pozzoli C, Poli E, et al. Effects of the Guide to Traditional Chinese Veterinary Medicine
macrolide antibacterial drug, tylosin, on TNBS- Small Animal Practice. Reddick, FL: Chi Institute
induced colitis in the rat. Pharmacology 2005; Press 2014:362.
74(3):135-142. 17. Xie H, Priest V. Xie’s Chinese Veterinary Herbology.
7. Mansfield CS, James FE, Craven M, et al. Remission Ames, Iowa: Blackwell Publishing 2010:132-133,
of histiocytic ulcerative colitis in Boxer dogs 392-393, 211-212.
correlates with eradication of invasive intramucosal 18. Bessant C. Chinese Herbs – Science or Tradition?
Escherichia coli. J Vet Intern Med 2009; 23(5):964- Integrative Veterinary Care Journal 2013; 3(2):48-
969. 49.
8. Plumb DC. Plumb’s Veterinary Drug Handbook 7th 19. Dubreuil, JD. Antibacterial and Antidiarrheal
Edition. Ames, Iowa: Wiley Blackwell 2011:916- Activities of Plant Products against Enterotoxinogenic
918. Escherichia coli. Toxins 2013; 5(11):2009-2041.
9. Lee, Justin. Metronidazole Risks. Clinician’s Brief 20. Iqbal M, Verpoote R, Henrie A, et al. Phytochemicals
digital edition; Sept 2014. as a potential source for TNF-α inhibitors.
Phytochemistry Reviews 2013; 12(1):65-93.
TCVM Practice/Client list for sale on Florida Gulf Coast.

World’s most beautiful beach!
Well-established solo TCVM practice with 1 assistant in the most affluent and alternative medicine
embracing area of the NW Florida Gulf Coast. Ongoing long term mutually beneficial relationships
with 2 conventional hospitals. Annual net over $140,000 in 2 days of appointments. Much more
potential. Sale price negotiable and includes owner financing and mentorship/assistance from one
of Chi Institute’s senior instructors and faculty, Dr Margaret Fowler.
Contact DrFowler@HolisticVetFL.com.

Case Series
The Therapeutic Effect of Acupuncture and Chinese

Herbal Medicine in 12 Dogs with Hyperadrenocorticism
Ronald Koh DVM, MS, Huisheng Xie DVM, PhD, May-li Cuypers DVM, DACVIM
ABSTRACT
Medical records of all dogs diagnosed with hyperadrenocorticism and treated with acupuncture and Chinese herbal medicine
between 2004 and 2013 were reviewed. Twelve dogs met inclusion criteria and 8/12 also had pre-treatment adrenocorticotrophic
hormone (ACTH) stimulation tests, urine cortisol-creatinine ratios (UCCR) and/or basal serum cortisol levels. Dogs had
received acupuncture for 5-42 months at the time of review. Mai Men Dong (Ophiophogon Formula®) alone or combined
with modified Jin Gui Shen Qi (Rehmannia 14®) or combined with modified Chai Hu Shu Gan (Liver Happy®), or modified
Xia Xiao Fang (Rehmannia 11®) alone were administered to all dogs for 12-45 months. No adverse side effects were seen.
Following treatment, polyuria, polydipsia and panting resolved in all 12 dogs. Lethargy improved more than 50% in all dogs.
Polyphagia resolved in 7/12 dogs and improved over 50% in 5/12 dogs. Skin and hair changes and distended abdomens
were more resistant, but improved more than 50% in 7/12 and 5/12 dogs respectively. Following a mean± standard deviation
(M±SD) of 21.9±13.35 months of treatment, 8/8 dogs studied had significantly (p<0.05) reduced post-ACTH stimulation
test results and 7/8 had returned to normal levels. All 5 dogs studied had significantly (p<0.05) improved UCCR following
20.8±13.27 months of treatment and 3 dogs had normal values. Basal cortisol levels returned to normal levels in 2/2 dogs
studied after a M±SD 8±4.24 months of treatment. Based on the treatment results in 12 dogs, acupuncture and Chinese herbal
medicine may be a safe and effective alternative to conventional medication and surgery for the treatment of some dogs with
hyperadrenocorticism.
Key words: Hyperadrenocorticism, Cushing’s disease, dogs, acupuncture, electroacupuncture, Chinese herbal medicine, Mai
Men Dong, Jin Gui Shen Qi, Xia Xiao Fang, Chai Hu Shu Gana
ABBREVIATIONS adenoma or carcinoma of the adrenal gland.1-3 Iatrogenic

Cushing’s syndrome has similar clinical signs and is
ACTH Adrenocorticotrophic hormone associated with the chronic overzealous administration
ALP Alkaline phosphatase of glucocorticoids with suppression of adrenal gland
ALT Alanine aminotransferase function. The clinical signs and diagnostic test results
M±SD Mean± standard deviation associated with hyperadrenocorticism vary based on the
TCM Traditional Chinese medicine duration and severity of the hypercortisolemia. Typical
TCVM Traditional Chinese veterinary medicine history, physical examination, clinicopathological and
UCCR Urine cortisol-creatinine ratio other diagnostic findings are outlined in Table 1. The
diagnosis can be challenging to confirm, but typical
Hyperadrenocorticism (Cushing’s disease) is a clinical parameters can lead to a strong suspicion.1-3
common endocrine disorder in dogs associated with Conventional medical and surgical treatments are
excessive secretion of glucocorticoid hormones by aimed at eliminating the increased secretion of cortisol.
the adrenal cortex.1-3 Approximately 85% of dogs Mitotane (o,p’-DDD [1,1-dichloro-2-(2-chlorophenyl)-2-
with hyperadrenocorticism have pituitary-dependent (4-chlorophenyl) ethane]) and trilostane (4α, 5α-epoxy-
hyperadrenocorticism with hyperplasia or neoplasia of the 17β-hydroxy-3-oxoandrostane-2α-carbonitrile), are
pituitary gland. The other 15% have a cortisol-secreting 2 medications currently recommended.4-12 Adverse
reactions, however, are common.4-7 In one study of
From: Louisiana State University School of Veterinary Medicine,
200 dogs receiving mitotane, approximately 25% of
Baton Rouge, LA (Koh), University of Florida College of Veterinary dogs exhibited one or more adverse effects during
Medicine, Gainesville, FL (Xie), Veterinary Mobile and Diagnostics, induction including: weakness, vomiting, anorexia,
Largo, FL (Cuypers)

Table 1: Signalment, history, physical examination, clinicopathological and other diagnostic test findings in dogs with
hyperadrenocorticism1,3,4
Signalment and Physical CBC, Serum Chemistry Other Clinicopathological and Diagnostic
History Examination and Urinalysis Tests
• Hypercortisolemia
• Neutrophilia
• Middle age or • Increased urine cortisol- creatinine ratio
• Obesity • Lymphopenia
older • Exaggerated increase in serum cortisol
• Pendulous • Monocytosis
• Polydipsia following ACTH stimulation test
abdomen • Eosinophilia
• Polyuria • No reduction of serum cortisol with low-dose
• Muscle atrophy • Lipemia
• Polyphagia dexamethasone suppression test
• Alopecia • Elevated serum ALT
• Lethargy • Lowered or unchanged serum cortisol with
• Thin skin • Elevated serum ALP
• Excessive high dose dexamethasone test (PDH)
• Comedones • Mild hyperglycemia
panting • Normal to increased plasma ACTH with PDH
• Calcinosis cutis • Hypercholesterolemia
• Bruising • Low plasma ACTH in adrenal tumors
• Hyperpigmentation • Decreased BUN
• Cool-seeking • Adrenal tumor visualized with abdominal
• Hepatomegaly • Glucosuria
behavior ultrasound, CT or MRI
• Hypertension • Urinary tract infection
• Pituitary tumor visualized on brain CT or
MRI
ALT =alanine aminotransferase; ALP= alkaline phosphatase; BUN= blood urea nitrogen; ACTH= adrenocorticotrophic
hormone; PDH= pituitary-dependent hyperadrenocorticism; CT= computerized tomography; MRI= magnetic resonance
imaging
diarrhea and ataxia.4 Diarrhea, vomiting and lethargy medicine can be safe and effective for many disorders,
have been reported in 63% of dogs receiving trilostane.5 when prepared and administered properly.19-21 Occasional
Hypoadrenocortical crisis and deaths associated with side effects are vomiting and diarrhea, which can usually
adrenal rupture and necrosis have also been reported be curtailed by reducing the dose. Typical TCVM patterns
with mitotane and trilostane treatment.1,6,7 Further, some associated with hyperadrenocorticism include: Kidney Yin
dogs with pituitary-dependent hyperadrenocorticism Deficiency initially leading to Kidney Qi Deficiency and
are not well controlled with trilostane.5-10 In some cases then resulting in Kidney Qi-Yin Deficiency.19,20 In later
surgical removal of adrenal and pituitary tumors can be stages of hyperadrenocorticism, Kidney Yang Deficiency
performed, but availability of equipment and trained often develops.
veterinary surgeons are limited and the procedures involve Although acupuncture and Chinese herbal medicine
risk and are expensive.13-14 Stereotactic radiosurgery and have been suggested for the long-term medical management
radiotherapy of pituitary tumors are also being performed of hyperadrenocorticism in dogs, scientific data regarding
at specialized centers.15 Radiosurgery is non-invasive and therapeutic effects is limited.18-21 At University of Florida
less risky than conventional surgery, but the availability Veterinary Medical Center (UFVMC), acupuncture and
and cost may be limiting factors for many clients. Chinese herbal medicine has been used to treat suspected
Originating 2,500 years ago in China, traditional and confirmed cases of hyperadrenocorticism in dogs for
Chinese medicine (TCM) especially acupuncture over 15 years. There are currently no English published
is becoming one of the most widespread forms of clinical studies of the treatment of hyperadrenocorticism
complementary and alternative medicine in the United in humans or animals with acupuncture and Chinese
State and Europe.16-21 In TCM and traditional Chinese herbal medicine. The purpose of this retrospective
veterinary medicine (TCVM), disease patterns can be study was to investigate the long-term effectiveness
identified (Bian Zheng) and are associated with imbalances of acupuncture and Chinese herbal medicines for the
(Excesses and Deficiencies) in the musculoskeletal and treatment of hyperadrenocorticism in dogs. Based on
nervous systems and internal organs of the body.17-22 anecdotal information, the hypothesis for this study was
Acupuncture and Chinese herbal medicines correct these that hyperadrenocorticism could be effectively managed in
imbalances so normal healing processes can be mobilized some dogs with acupuncture and Chinese herbal medicine
to restore health. People are becoming more interested without adverse side effects.
in effective alternative treatment options with fewer side
effects and risks than those of conventional medical or MATERIALS AND METHODS
surgical treatments.16 Acupuncture has rare side effects Medical records of 20 dogs diagnosed with
that include bleeding, bruising and local infection in hyperadrenocorticism and treated with a combination of
the area where the needle is inserted.17 Chinese herbal acupuncture and Chinese herbal medicine at the UFVMC

between 2004 and 2013 were reviewed. Inclusion lethargy (n=11), excessive panting (n=10), cool-seeking
criteria included: 1) middle aged and older dogs, 2) a behavior (n=11), exercise intolerance (n=9), weight gain
typical history and physical examination, complete blood (n=8), and pendulous abdomen (n=7) (Table 2). Thin
count, serum biochemistry and urinalysis findings for skin, alopecia and hyperpigmentation occurred in 9 dogs
hyperadrenocorticism, 3) acupuncture and Chinese herbal and calcinosis cutis in 1 dog. All 12 dogs had moderate
medicine treatment of the disease for at least 5 months to marked elevations of serum alanine aminotransferase
and 4) Follow-up information available about resolution (ALT) and alkaline phosphatase (ALP). Other abnormal
or improvement of clinical signs. Cases of suspected findings included: mild leukocytosis (7 dogs), mild
hyperadrenocorticism were excluded if: 1) Records were hypercholesterolemia (4 dogs) and marked hepatomegaly
incomplete as to clinical signs, treatments or outcomes, on abdominal radiographs (5 dogs). The duration of
2) Treatment was less than 5 months and 3) No follow- clinical signs prior to treatment ranged from 2-8 months
up information was recorded or could be collected from with a M±SD of 4.75±1.71 months (Table 2).
clients. Concurrent illnesses were present in 8/12 dogs and
Two veterinarians certified in veterinary acupuncture included: 2 dogs with hypothyroidism, 2 dogs with chronic
and Chinese herbal medicine treated all patients. Dry renal failure, 1 dog with atopy and pyoderma, 1 dog with
needle acupuncture, electroacupuncture and aqua- epilepsy, 1 dog with keratoconjunctivitis sicca, cataracts,
acupuncture were performed during acupuncture Grade III/VI left apical systolic heart murmur and urinary
sessions. All dogs received one or more Chinese herbal tract infection, and 1 dog with diabetes mellitus and
medicines.19-21 Clinical response to treatment of dogs was urinary tract infection (Table 2).
based on the history and physical examination findings A presumptive conventional diagnosis of
during subsequent visits, and follow-up information via hyperadrenocorticism had been made on the 12 dogs based
telephone. Dogs with resolution of clinical signs were on the typical historical and physical examination findings,
classified as 100% improvement. Moderate improvement but 8 dogs also had testing done more specifically to
was characterized by greater than 50% (>50%) support the diagnosis of hyperadrenocorticism. All 8 dogs
improvement, but not resolution of clinical signs. Dogs had ACTH stimulation tests, 5/8 dogs also had UCCR and
that had between 50%-30% improvement of clinical 2/8 dogs had basal serum cortisol assays (Table 2). Based
signs were considered to have mild improvement. Dogs on the TCVM pattern diagnostic system, 8/ dogs had Yin
considered to have less than 30% improved were classified Deficiency, 2 dogs had Qi and Yin Deficiency, 1 dog had
as unimproved. Yin Deficiency with Liver Qi Stagnation, and 1 dog had
Although not an inclusion criteria, 8 dogs included in Yin and Yang Deficiency on presentation (Table 2).
the study had pre-treatment adrenocorticotrophic hormone Acupuncture treatment entailed the insertion of
(ACTH) stimulation testsa, urine cortisol-creatinine sterile 13 mm or 25 mm acupuncture needlesf, based on the
ratiosb (UCCR) and/or basal serum cortisol concentration size of patient and the location of the acupuncture points
assaysc. The same 8 dogs had post treatment testing every treated. Generally 8-14 acupuncture points were treated.
3-6 months after commencement of acupuncture and Needles were left in place for 10-20 minutes. Electrical
Chinese herbal medicine. The same laboratory ran all stimulationg for 10 minutes was administered to 6/12 dogs
tests. Accepted normal values were a serum cortisol level at different sessions. Aqua-acupuncture with injection of
post-ACTH stimulation test of 6-17 μg/dld, UCCR of 15 diluted vitamin B12h with salinei at acupuncture points was
or lessd, and basal serum cortisol concentration of 0.5-6 also performed in 10/12 dogs at different sessions. Aqua-
μg/dld. acupuncture was performed at acupoints BL-23, KID-3,
Comparisons of pre and post-ACTH serum cortisol SP-6, SP-9, SP-8 ST-36, in addition to acupoints selected
concentrations and UCCR on individuals before and based on pattern diagnosis. Initially, all 12 dogs received
after treatment were performed by boxplots. The mean ± acupuncture therapy every 2-4 weeks. The time between
standard deviation (M±SD) of all dogs before and after sessions was progressively lengthened to every 2-6 month
treatment was analyzed using the Student paired t test. intervals as the severity of clinical signs decreased. The
Differences were considered significant at p values equal number of acupuncture treatments per dog ranged from
to or less than (<) 0.05. Standard statistical softwaree was 6-21, with a M±SD of 12.42±4.7 treatments. Acupuncture
used for all computations. treatment duration ranged from 5-42 months with a M±SD
of 19.41±11.74 months (Table 3).
RESULTS Modified Mai Men Dong (Ophiophogon Formula®j)
Twelve dogs, 7-14 years old (average age 10.5 alone was administered to 8 dogs with Yin Deficiency
years), met the inclusion criteria (Table 2). There were (Tables 2-4). Modified Mai Men Dong (Ophiophogon
10 neutered females and 2 neutered males with 7 pure Formula®j) and modified Chai Hu Shu Gan (Liver Happy®)
breed and 5 mixed breed dogs represented (Table 2). j
) were administered to 1 dog with a combination pattern
The most common initial clinical signs reported were of Yin Deficiency and Liver Qi Stagnation (Tables 2-5).
polyuria (n=12), polydipsia (n=12), polyphagia (n=9), Modified Mai Men Dong (Ophiophogon Formula®j)

Table 2: Signalment, primary clinical signs, duration of signs prior to treatment, concurrent disease, TCVM pattern and
special
diagnostic tests of 12 dogs with hyperadrenocorticism
Clinical Duration Concurrent Initial TCVM Diagnostic Tests for
Case Signalment
Signs of Signs Disease Pattern(s) Hyperadrenocorticism
10 year old,
PU, PD, PP, ACTH Stimulation test
female spayed,
1 L, EP, CS, 5 months Hypothyroidism UCCR
Labrador Yin Deficiency
EI, WG, SC Basal serum cortisol
Retriever
11 year old, PU, PD, PP, ACTH Stimulation test
Qi-Yin
2 female spayed, L, EP, CS, 6 months None recorded UCCR
Deficiency
Dachshund EI, WG, PA Basal serum cortisol
10 year old, PU, PD, L,
Chronic renal ACTH Stimulation test
3 female spayed, EP, CS, EI, 4 months Yin Deficiency
failure UCCR
mixed breed SC
PU, PD, PP,
7 year old, Yin Deficiency
L, EP, CS, ACTH Stimulation test
4 female spayed, 8 months Hypothyroidism and Liver Qi
EI, WG, PA, UCCR
mixed breed Stagnation
SC
10 year old, PU, PD, PP,
ACTH Stimulation test
5 female spayed, EP, CS, WG, 4 months Atopy, pyoderma Yin Deficiency
UCCR
mixed breed PA, SC
11 year old,
PU, PD, PP,
male neutered,
6 L, CS, EI, 7 months Epilepsy Yin Deficiency ACTH Stimulation test
American
WG
Eskimo

Chronic renal
7 female spayed, EP, CS, EI, 4 months Yin Deficiency ACTH Stimulation test
failure
Chinese crested PA, SC
Keratoconjunctivitis
sicca, cataracts,
Grade III/VI left
8 female spayed, L, EP, CS, 6 months Yin Deficiency ACTH Stimulation test
apical systolic heart
Pekingese PA, SC
murmur, urinary
tract infection
female spayed, L, EP, CS, Qi-Yin
9 4 months None recorded None performed
Yorkshire EI, WG, PA, Deficiency
Terrier SC
Yin-Yang
10 female spayed, L, EI, WG, 2 months None recorded None performed
Deficiency
Dachshund PA, SC
11 female spayed, EP, CS, EI, 3 months None recorded Yin Deficiency None performed
mixed breed WG
11 year old, PU, PD, PP, Diabetes mellitus,
12 male neutered, L, EP, CS, 4 months urinary tract Yin Deficiency None performed
mixed breed EI, SC infection
Mean±SD 9.67±1.23 N/A 4.75±1.71 N/A N/A N/A
PU= polyuria, PD= polydipsia, PP= polyphagia, L= lethargy EP= excessive panting, CS= cool seeking, EI= exercise
intolerance, WG= weight gain, PA= pendulous abdomen, SC= skin changes; N/A= not applicable

Table 3: Signalment, number and duration of acupuncture treatments, duration of Chinese herbal medicine treatment and
clinical outcomes of 12 dogs with hyperadrenocorticism
Duration of Duration of
Number of
Acupuncture Chinese Herbal Herbal Clinical Signs
Case Signalment Acupuncture
Treatment** Medicine Medicine** Resolved**
treatments
(months) (months)
10 year old, female spayed, Ophiopogon PU, PD, PP, L, EP,
1 14 11
Labrador Retriever Formula 14 EI, WG, SC
11 year old, female spayed, PU, PD, PP, L, EP,
2 6 5 Rehmannia 11
Dachshund 12 EI, WG, PA
10 year old, female spayed, Ophiopogon
3* 21 39 L, EP, EI, SC
mixed breed Formula 45
Ophiopogon
4 7 24 Formula,
mixed breed 36 EI, WG, PA, SC
Liver Happy
10 year old, female spayed, Ophiopogon PU, PD, PP, EP,
5 9 25
mixed breed Formula 30 WG, PA, SC
11 year old, male neutered, Ophiopogon PU, PD, PP, L, EI,
6* 15 42
American Eskimo Formula 46 WG
10 year old, female spayed, Ophiopogon
7 17 18 L, EP, EI, PA, SC
Chinese crested Formula 18
8 year old, female spayed, Ophiopogon PU, PD, PP, L, EP,
8 12 11
Pekingese Formula 20 PA, SC
9 12 Rehmannia 11
Yorkshire Terrier 12 18 EI, WG, PA, SC
Ophiopogon
9 year old, female spayed, PU, PD, PP, L, EI,
10 10 Formula,
Dachshund 8 26 WG, PA, SC
Rehmannia 14
10 year old, female spayed, Ophiopogon PU, PD, L, EP, EI,
11 24
mixed breed 18 Formula 24 WG
11 year old, male neutered, Ophiopogon
12 12 L, EP, EI, SC
mixed breed 10 Formula 17
Mean±
9.67±1.23 12.42±4.7 19.41±11.74 N/A 25.5±11.53 N/A
SD
* Clients indicated the dog had died at the final communication (all others were alive); ** final client communication time
after onset of treatment; PU= polyuria, PD= polydipsia, PP= polyphagia, L= lethargy EP= excessive panting, CS= cool
seeking, EI= exercise intolerance, WG= weight gain, PA= pendulous abdomen, SC= skin changes.
Table 4: Ingredients of the Chinese herbal medicine modified Mai Men Dong (Ophiopogon Formula®f) and their actions19
Pin Yin Name English Name Actions
Mai Men Dong Ophiopogon Nourishes Yin
Lu Gen Phragmites Clears Heat, promotes Body Fluids
Zhi Mu Anemarrhena Nourishes Yin, clears Heat
Tian Hua Fen Trichosanthes Clears Heat, promotes Body Fluids
Bei Shan Shen Glehnia Nourishes Yin
Zhu Ye Bambusa Clears Heat
Ge Gen Pueraria Promotes Body Fluids
Wu Mei Mume Astringently consolidates
Huang Qin Scutellaria Clears Heat
Yu Li Ren Prunus Moves Blood
Shan Zha Crataegus Resolves Food Stagnation
Shen Qu Massa Fermentata Resolves Food Stagnation

Table 5: Ingredients of the Chinese herbal medicine modified Chai Hu Shu Gan (Liver Happy®f) and their actions19
Xiang Fu Zi Cyperus Soothes Liver, resolves Stagnation
Qing Pi Citrus Moves Qi, soothes Liver, resolves Stagnation
Bai Shao Yao Paeonia Soothes Liver, nourishes Liver Yin and Blood
Chen Pi Citrus Dries Dampness, moves Qi
Bo He Mentha Moves Qi
Chai Hu Bupleurum Soothes Liver
Mu Dan Pi Moutan Cools Blood, clears Heat, dissipates Stagnation
Zhi Zi Gardenia Clears Heat
Dang Gui Angelica Moves Blood
Gan Cao Glycyrrhiza Harmonizes the formula
Table 6: Ingredients of the Chinese herbal medicine modified Jin Gui Shen Qi (Rehmannia 14®f) and their actions19
Sheng Di Huang Rehmannia Nourishes Yin and Jing
Shan Yao Dioscorea Tonifies Qi and Jing
Bai Shao Yao Paeonia Nourishes Liver Yin and Blood
Huang Qi Astragalus Tonifies Qi
Zhi Mu Anemarrhena Clears Heat, promotes Body Fluids and Yin
Huang Bai Phellodendron Clear Heat, nourishes Yin
Wu Wei Zi Schisandra Consolidates and nourishes Lung Yin
Mai Men Dong Ophiopogon Nourishes Yin
Fu Ling Poria Drains Damp, strengthens Spleen
Ze Xie Alisma Drains Damp
Gui Zhi Cinnamomum Warms and tonifies Kidney Yang
Fu Zi Aconite Warms and tonifies Kidney Yang
Shan Zhu Yu Cornus Nourishes Yin
Table 7: Ingredients of the Chinese herbal medicine modified Xia Xiao Fang (Rehmannia 11®f) and their actions19
Huang Qi Astragalus Tonifies Qi
Rou Gui Cinnamomum Warms and tonifies Kidney Yang
Shu Di Huang Rehmannia Nourishes Kidney Jing and Yin
Shan Yao Dioscorea Tonifies Qi
Shan Zhu Yu Cornus Nourishes Yin
Fu Ling Poria Drains Damp, strengthens Spleen
Ze Xie Alisma Drains Damp
Fu Pen Zi Rubus Astringently consolidates
Jin Ying Zi Rosa Astringently consolidates
Sang Piao Xiao Mantidis Astringently consolidates

and modified Jin Gui Shen Qi (Rehmannia 14®j) were (17%) respectively. At the fifth re-evaluation (median of
administered to 1 dog with a combination pattern of Yin 18 weeks) further improvement of skin and distended
and Yang Deficiencies (Tables 2-4 and 6). Modified Xia abdomens had increased to 7/12 dogs (58%) and 5/12
Xiao Fang (Rehmannia 11®j) alone was administered to 2 dogs (42%) respectively. At a median time of 2 years of
dogs with a combination pattern of Qi and Yin Deficiencies treatment 9/12 dogs (75%) were described by caretakers
(Tables 2, 3 and 7). Chinese herbal medicine treatment as having resolution of clinical signs, 2/12 dogs (17%)
duration ranged from 12-46 months with a M±SD of had overall moderate improvement, and 1 dog (8%) had
25.5±11.53 months at the time of review. only slight improvement (30%-50%). Long-term follow-
No adverse side effects were noted with either the up information on 10/12 dogs was obtained from phone
acupuncture or Chinese herbal medicine treatments. An calls to clients. After 18 and 24 months respectively, no
overview of resolution or improvement of clinical signs contact could be made for follow-up with clients with
on re-evaluations at median time periods after the onset Cases 7 and 11. Cases 2 and 6 had succumbed to renal
of therapy is outlined in Table 3. At the first re-evaluation and cardiac failure respectively after 12 and 46 months of
(median of 3 weeks) after initiation of acupuncture treatment for hyperadrenocorticism. The 8 surviving dogs
and Chinese herbal medicine, 12/12 dogs (100%) had where client contact could be made were still receiving
moderate improvement (>50%) of polyuria, polydipsia, acupuncture every 3-6 months and daily Chinese herbal
panting and activity levels. At the second re-evaluation medicine. At the time of contact, the mean survival time
(median of 6 weeks), 5/12 dogs (42%) had resolution for the 10 dogs had been M±SD of 31±2 months.
of polyuria, polydipsia and excessive panting, and 6/12 The ACTH stimulation test was used to monitor the
dogs (50%) showed moderate improvement (<50%) in response to acupuncture and Chinese herbal medicine
polyphagia and skin and hair coat condition. By the forth in 8/12 dogs (67%) in this study (Table 8). Following a
re-evaluation (median of 12 weeks), all 12 dogs had M±SD of 21.9 ± 13.35 months of acupuncture and Chinese
resolution of polyuria, polydipsia and excessive panting. herbal medicine, all 8 dogs had significantly reduced post-
At this time there was also resolution of polyphagia in ACTH serum cortisol concentration (p<0.05) compared
6/12 dogs (50%) and mild improvement (>50%) of the to pre-treatment values. Further 7 dogs had post-ACTH
distended abdomens in 6/12 dogs (50%) and 2/12 dogs stimulation test results that had returned to accepted
Table 8: Data of 8 dogs monitored with post-ACTH stimulation tests following acupuncture and Chinese herbal medicine
treatments
Case Post-ACTH cortisol before Length of Post-ACTH cortisol
Signalment
number treatment (μg/dl) treatment (months) after treatment (μg/dl)
10 year old, female spayed,
1 27.3 11 15.1*
Labrador Retriever
2 38.6 5 19.4*
Dachshund
3 28.9 39 13.9*
mixed breed
4 15.6 24 6.2*
mixed breed
5 20.1 25 15.3*
mixed breed
11 year old, male neutered,
6 29.5 42 8.4*
American Eskimo
7 23.5 18 11.2*
Chinese crested
8 21.2 11 14.5*
Pekingese
Mean ±
9.6 ± 1.41 year old 25.6 ± 7.09 21.9 ± 13.35 13.1 ± 4.21*
SD
Length of treatment was measured from the beginning of treatment until the dog died or records reviewed for the study;
*significant decrease (p<0.05) after treatment compared to pre-treatment values; Seven of the eight dogs had normal post-
ACTH stimulation serum cortisol levels of 6-17 ug/dld after treatment
Figure 1: Post ACTH serum cortisol concentrations of 8 dogs before treatment (blue bars) and after treatment of 11, 5, 39,
24, 25, 42, 18 and 11 months (Cases 1-8) respectively of acupuncture and Chinese herbal medicine (red bars); *= significant
reduction after TCVM treatment (p<0.05)
Figure 2: Mean ± standard (M±SD) deviation of ACTH stimulation test results in 8 dogs before treatment (blue bar) and after
treatment of a M±SD of 21.9 ±13.35 months of acupuncture and Chinese herbal medicine (red bar); *= significant reduction
after TCVM treatment (p<0.05)

normal values range (serum cortisol levels 6-17 ug/dl) hyperadrenocorticism include gluoconeogenesis,
(Table 8, Figures 1 and 2). immunosuppression, anti-inflammatory responses and
The UCCR was used to monitor 5/12 dogs (42%) in protein catabolism.22 Uncontrolled hyperadrenocorticism
this study (Table 9). All 5 dogs had significantly (p<0.05) markedly affects the quality of life of both dogs and clients
improved UCCR following a M±SD of 20.8 ± 13.27 and can lead to a number of complications including
months of acupuncture and Chinese herbal medicine infections, life-threatening thromboembolism, generalized
treatments. Further, 3/5 dogs (60%) had UCCR within catabolism and muscle fibrosis.1-3
normal values range (less than 15) (Table 9, Figure 3). From a TCVM perspective hyperadrenocorticism is a
The basal serum cortisol levels were monitored Lower Burner (Xia Jiao) thirsting and wasting disorder as
on 2/12 dogs (17%) (Table 10). The basal serum exemplified by the clinical signs of polydipsia, polyuria,
cortisol concentrations before and after treatment were polyphagia and loss of muscle mass.20 Early stages of the
significantly (p<0.05) different. In both dogs basal cortisol disease may be a mixed Excess and Deficiency disorder.
levels returned to normal values range (0.5-6 μg/dl) after Liver Qi Stagnation and Lung and Stomach Yin Deficiency
a M±SD of 8±4.24 months of acupuncture and Chinese cause Heat, as evidenced by panting (especially at night)
herbal medicine treatments. and cool-seeking behavior. A purple tongue and wiry
pulse is often found in dogs with Liver Qi Stagnation and
DISCUSSION a red tongue and a rapid, thready pulse in dogs with Yin
From a conventional perspective the wide spread Deficiency.20 Global Yin and Qi Deficiency follow and
effects of excess glucocorticoids in animals with affected dogs also develop lethargy, exercise intolerance
Table 9: Data of 5 dogs monitored with urine cortisol-creatinine ratio (UCCR) following acupuncture and Chinese herbal
medicine treatments
UCCR Length of UCCR
Case number Signalment
before treatment treatment (months) after treatment
1 10 year old, female spayed, Labrador 20 11 12*
Retriever
2 11 year old, female spayed, Dachshund 19 5 13*
3 10 year old, female spayed, mixed 22 39 18*
breed
4 7 year old, female spayed, mixed breed 26 24 19*
5 10 year old, female spayed, mixed 29 25 14*
breed
Mean ± SD 9.6 ± 1.52 year old 23.2 ± 4.20 20.8 ± 13.27 15.2 ± 3.11*
*significant decrease (p<0.05) after treatment compared to pre-treatment values; three dogs had normal ratios of less than 15d
after treatment
Table 10: Data of 2 dogs monitored with basal serum cortisol concentration following acupuncture and Chinese herbal
medicine treatments
Case Serum cortisol before Length of treatment Serum cortisol after
Signalment
number treatment (μg/dl) (months) treatment (μg/dl)
10 years old, female spayed,
1 9.5 11 6.0*
Labrador Retriever
11 years old, female spayed,
2 7.5 5 3.0*
Dachshund
Mean ± SD 10.5 ± 0.71 years old 8.5 ± 1.41 8 ± 4.24 4.5 ± 2.83
*significant (p<0.05) reduction of basal serum cortisol concentrations after treatment compared to pre-treatment values; both
dogs in normal cortisol levels between 0.5-6 μg/dld after treatment
and loss of muscle tone.19,20 Once Kidney/Spleen Yang hyperadrenocorticism include: ST-36, SP-6, SP-9, SP-
Deficiency develops the liver is palpably enlarged, the hair 10, BL-17, BL-23, BL-26, KID-3, KID-7, LIV-2, LIV-3,
coat and skin are thin, the ears, back and limbs are cool and CV-4 and CV-6.18,20 Typical acupoints treated for Yang
the dog has warm seeking behavior. 19,20 At this stage the Deficiency hyperadrenocorticism include: ST-36, SP-6,
tongue is pale and swollen and pulses are weak deep and BL-23, KID-7, KID-13, GV-3, GV-4, CV-4, CV-6 and
slow.19,20 In the study, 8/12 dogs (67%) had Yin deficiency. moxibustion at CV-8.20 The general acupoints that were
These dogs showed signs of panting, red tongue, fast treated in the 12 dogs of the study included: GV-20 (for
pulses, red skin, and cool seeking. A pattern diagnosis of calming), BL-23, KID-7, SP-6, SP-9, ST-36 and LIV-3,
a combination of Qi and Yin Deficiency was diagnosed in in addition to the acupoints that were selected based on
2/12 dogs (17%) based on lethargy, exercise intolerance, pattern differentiation. Acupoints, such as GV-14, LI-
or pale tongue, in addition to Yin Deficient signs. A 4, LI-11, Er-jian were added if the dog had Heat in the
combination of Yin deficiency with Liver Qi Stagnation body that needed to be cleared. For electroacupuncture,
was made in 1 dog (8%) based on clinical signs of Yin the connections of leads also varied among dogs for each
Deficiency along with a wiry pulse and irritable behavior. session, but in general lead included: BL-23 bilaterally,
A combination of Yin and Yang Deficiency was made in ST-36 bilaterally, and SP-6 to KID-3 on each limb.
1 dog (8%) with a pale tongue, slow pulse, lethargy and Like acupuncture, the selection of the appropriate
cool body. Chinese herbal medicine is based on the underlying TCVM
Acupuncture points treated vary with the TCVM pattern. Modified Mai Men Dong (Ophiopogon Formula®)
pattern. Therefore all dogs with a conventional diagnosis nourishes Yin, clears Heat and promotes Body Fluids so is
of hyperadrenocorticism will not receive the same often used in Yin Deficient hyperadrenocorticism (Table
treatment, as they do in conventional medicine. As 4).19-21 Modified Chai Hu Shu Gan (Liver Happy®)) is used
typical in TCVM practice, acupuncture was performed to soothe Liver and resolve Qi Stagnation, so may also
at different acupuncture points in different dogs in this be combined with modified Mai Men Dong (Ophiopogon
study. Also different acupuncture points were treated at Formula®) in TCVM patterns where Liver Qi Stagnation is
subsequent treatment sessions for the same dog, based on apparent (Table 5). Modified Xia Xiao Fang (Rehmannia
the individual’s current pattern and response to previous 11®) tonifies Qi and nourishes Yin so may be prescribed
treatments. Typical acupoints for hyperadrenocorticism in Qi/Yin Deficient hyperadrenocorticism (Table 7).
with Yin Deficiency include: BL-23, BL-52, KID-3, Modified Jin Gui Shen Qi (Rehmannia 14®) is used to
KID-7, KID-6, KID-10, SP-6, SP-9, SP-8 and LIV- warm and tonify Kidney Yang while also tonifying Kidney
3.18,20 Typical acupoints treated for Qi-Yin Deficiency Yin so may also be combined with modified Mai Men
Figure 3: The urine cortisol-creatinine ratio of 5 dogs before treatment (blue bars) and after treatment evaluated after 11, 5,
39, 24 and 25 months respectively (Cases 1-5) of acupuncture and Chinese herbal medicine treatment (red bars); *
= significant reduction after treatment (p<0.05)

Dong (Ophiopogon Formula®) in Yin/Yang Deficiency compared to the 11-42 months of treatment in the other
combination patterns (Tables 4 and 6) .19-21 All 12 dogs dogs. The UCCR was significantly (p<0.05) lower in all 5
in the study were administered the correct Chinese herbal dogs evaluated, but remained slightly elevated above the
medicine based on their pattern differentiation (Table 3). normal 15 or less range in 2 dogs (ratios of 18 and 19 in
Typically these Chinese herbal medicines are administered Case number 3 and 4 respectively, Table 9). Both of those
at a dose of 0.5g/10-20 pounds body weight twice daily dogs however had normal post ACTH stimulation serum
for 6 months. In this study, the herbal treatment continued cortisol levels (Case numbers 3 and 4, Table 11). Case
beyond 6 months (12-46 months) with no adverse effects number 2 that still had an elevated post ACTH stimulation
because of the need for long term control of a disease that serum cortisol level following 5 months of treatment had
can not be cured.9,3,20 a normal UCCR and basal cortisol levels after treatment
Polyuria, polydipsia, polyphagia, lethargy and (Tables 8-10).
excessive panting are the most common initial signs of While monitoring tests give an objective indication
hyperadrenocorticism, as evidenced by all 12 dogs in the as to what is occurring inside the body, they do not
current study, and affect the quality of life for both the give any indications as to how well the patient appears
animal and caretaker.1-3 At a median of 3 weeks after the physically or clinically, which may be a more reliable
onset of acupuncture and herbal treatment all dogs had a indicator of the patient’s condition.1-3 Overall acupuncture
greater than 50% improvement of polyuria, polydipsia, and Chinese herbal medicine treatment of the 12 dogs
panting and activity levels and by 3 months polyuria, with hyperadrenocorticism in the study resulted in a
polydipsia and panting had resolved, with no adverse side 100% improvement of polyuria, polydipsia and panting, a
effects (Tables 3 and 11). Such improvements can resolve greater than 50% improvement in activity and resolution
many quality of life issues such as urinary accidents in the of polyphagia in 58% of dogs (Table 3). Skin and hair
house and panting at night which disturb clients that sleep changes improved greater than 50% in 58% of dogs and
with their dogs. In additions, clients feel less stressed, 42% of dogs had improvement in distended abdomens.
when their pets have improved activity levels and are less The inclusion criteria of long-term follow-up did bias the
lethargic. results however, as only clients that saw improvement
All 8 dogs in the study that were monitored by ACTH would have returned for further acupuncture treatments
stimulation tests had a significant (p<0.05) reduction of and administered herbal capsules twice daily.
post ACTH stimulation serum cortisol levels following Whether from a conventional or TCVM
treatment. Further in 7/8 dogs (87.5%), the post treatment perspective, the primary goal of treatment of dogs
levels had returned to normal (Table 8). The one dog that with hyperadrenocorticism is aimed at controlling and
had post ACTH serum cortisol levels that was still above achieving resolution of clinical signs associated with
the normal range had only been treated for 5 months hypercortisolemia rather than curing the disease.3,9,20
Table 11: Percent improvement of clinical signs on re-evaluations; median re-evaluation periods of 3-18 weeks of treatment
of 12 dogs.
Number of Dogs and Percentage Improved at Median Re-evaluation Periods

Clinical Signs
25.5±11.53
3 weeks 6 weeks 12 weeks 18 weeks
months
5 dogs 100%
Polyuria 12 dogs >50% 12 dogs 100% 12 dogs 100% 12 dogs 100%
7 dogs >50%
5 dogs 100%
Polydipsia 12 dogs >50% 12 dogs 100% 12 dogs 100% 12 dogs 100%
7 dogs >50%
5 dogs 100%
Panting 12 dogs >50% 12 dogs 100% 12 dogs 100% 12 dogs 100%
7 dogs >50%
Activity 12 dogs >50% 12 dogs >50% 12 dogs >50% 12 dogs >50%
6 dogs 100% 7 dogs 100%
Polyphagia No change 6 dogs >50%
6 dogs >50% 5 dogs >50%
6 dogs <50% 7 dogs >50%
Skin/Hair No change 6 dogs <50%
6 dogs no change 5 dogs <50%
Distended 2 dogs <50% 5 dogs >50%
No change No change
abdomen 10 dogs no change 7 dogs <50%

It has been reported that 80-86% of dogs with 30 minutes, 3 times a week for 3 weeks. At the end of 3
hyperadrenocorticism are well controlled with mitotane, week experimental treatment period, all 4 dogs treated at
but 46% relapse.23 Reports on the efficacy of trilostane ST-36 had significantly increased post ACTH stimulation
vary from 62-100% and 69% with ketoconazole.5,8,11,23,24 cortisol concentrations (p<0.05) and the 4 dogs with sham
The adverse side effects of both mitotane and trilostane acupuncture showed no significant change. The authors
are well-documented and observed in approximately 25% concluded that electroacupuncture of ST-36 can regulate
of treated dogs.25 Therefore treatment with mitotane or the hypothalamus-pituitary-adrenal axis to restore function
trilostane involves frequent monitoring by the veterinarian in experimentally created adrenocortical hypofunction in
and a vigilant caretaker who is willing to closely monitor dogs.
the dog for signs of adverse side effects.8 There were no This is the first study of the outcome of dogs treated
side effects associated with the acupuncture and Chinese with acupuncture and Chinese herbal medicine from
herbal medicine in the 12 dogs with hyperadrenocorticism hyperadrenocorticism. Acupuncture and Chinese herbal
in the current study and there was greater than 50% medicine resolved or improved some or all of the clinical
improvement of polyuria, polydipsia and panting in just 3 signs of hyperadrenocorticism and significantly decreased
weeks with resolution in 3 months. basal and post-ACTH cortisol levels and UCCR in the 12
The exact mechanisms of the therapeutic effect dogs in this study. Although further studies are needed
of acupuncture and Chinese herbal medicine in with greater numbers of dogs, the study hypothesis
hyperadrenocorticism are unclear. It has been suggested was supported by the results. Therefore treatment with
that acupuncture promotes natural functional homeostasis acupuncture and modified Mai Men Dong (Ophiopogon
and possesses such effects as modulating neurotransmitter Formula®), modified Chai Hu Shu Gan (Liver Happy®))
function in the central nervous system as well as modified Xia Xiao Fang (Rehmannia 11®), and/or modified
adjusting hormonal imbalances.26,27 A well-known action Jin Gui Shen Qi (Rehmannia 14®) may be an alternative
of acupuncture is that it induces release of endogenous to conventional medication and surgery for the control of
endorphins resulting in analgesia.28 The endogenous opioid hyperadrenocorticism in some dogs.
system and pituitary-adrenal axis have a close physiological
link.28 One therapeutic mechanism of acupuncture and Footnotes
Chinese herbal medicine for hyperadrenocorticism a. Nichols Advantage ACTH Assay, Nichols Institute
may be associated with feedback inhibition of pituitary Diagnostics, Bad Vilbel, Germany
ACTH or hypothalamic corticotrophin releasing factor b. DPC Corporation, San Diego, California, USA
by beta-endorphin following acupuncture.28,29 An inverse (UCCR measured in Metrolab Autoanalizer Merck,
relationship between cortisol and endorphin was reported Germany)
as early as the 1980s.29 Taylor et al found that cortisol levels c. Coat-a-Count cortisol assay, Diagnostic Products
decreased below the basal level in response to infusion of Corp, Los Angeles, CA
beta-endorphin in normal humans and proposed that beta- d. University of California Davis School of Veterinary
endorphin might suppress cortisol release. Medicine Teaching Hospital data base
There is a growing body of studies that support e. SPSS, version 14.0, SPSS Inc., Chicago, IL 60606
acupuncture associated changes in the hypothalamic f. Tai Chi, Lhasa OMS, Inc., Weymouth, MA
pituitary axis with serum cortisol increasing or decreasing g. Donghua Electronic Instrument Factory, Beijing
dependent on the clinical situation. 30-34 Two studies 100083, PR China
looked at the effect of electroacupuncture on humans h. Vedco, Inc., St. Joseph, MO 64507
with increased levels of stress related serum cortisol due i. Hospira, Inc., Lake Forest, IL 60045
to orthopedic surgery and pain associated with chronic j. Dr. Xie’s Jing Tang Herbal Inc., Reddick, FL
osteoarthritis in the knee. Both studies demonstrated
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Endocrinol 1983;103(4):469-472. iatrogenic Cushing’s syndrome in dogs with
34. Lee SC, Yin SJ, Lee ML et al. Effects of acupuncture electroacupuncture stimulation of stomach 36. Am J
on serum cortisol level and dopamine beta- Chin Med 1991;19(1):9-15.
Alt-Vet The Revolutionary Pet Care and

Longevity Solution explores the TCVM
therapies that are available for companion
animals
Preface by Dr Huisheng Xie
Contains real life stories of pet owners that

were searching for any hope of saving their
pet, reducing their pain, and increasing
their quality of life.
Alt-Vet offers chapters on pet acupuncture,

massage, healing diets, prevention care
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Available in bookstores, amazon.com &

www.mitsievargas.com
About the Author Her love for Traditional Chinese Veterinary Medicine
Dr. Vargas received her DVM from Tuskegee University coupled with her desire to educate the public manifested
in 1994. In 1996 she established her AAHA accredited in her book Alt-Vet: The Revolutionary Pet Care and
Orchid Springs Animal Hospital in Winter Haven, FL. In Longevity Solution. In her practice she experienced
2009 she started her studies at Chi Institute, eventually unexpected successes treating terminal diseases and
garnering certifications in acupuncture, Chinese herbal conditions that were poorly responsive to allopatic
medicine, food therapy and Tui-na. She is pursuing her medicine. The hope and emotional response of the owners
Master’s Degree in TCVM at Chi Institute, FL where that came to her as the last option for their beloved pets
she works as a teacher assistant and as a lecturer for Chi ignited a desire to spread the word about TCVM as a valid
Latin America. In addition, she has worked as a TCVM medical option. The book is full of advice on responsible
consultant for Disney Animal Kingdom. Dr. Vargas pet ownership and holistic pet care. Food therapy and
was honored by the American Veterinary Foundation as Acupuncture are heavily featured as Dr. Vargas considers
“America’s Favorite Veterinarian” in 2015. them the basis of preventative medicine for pets.
She lives in sunny Winter Haven, Florida, with her
Dr. Vargas is also a certified veterinary journalist and a husband, two daughters and a menagerie of pets.
prolific author of both scientific and motivational articles.

Review
Review of Evidence-Based Clinical and

Experimental Research on the Use of Acupuncture
and Chinese Herbal Medicine for the Treatment or
Adjunct Treatment of Cancer
Huisheng Xie DVM, MS, PhD, Betsy Hershey DVM, DACVIM (Oncology), Aituan
Ma DVM, MS, PhD
ABSTRACT
Traditional Chinese Medicine includes acupuncture and herbal medicine. Clinical evidence indicates that acupuncture and
herbal medicine can be beneficial in the treatment of cancers in people and animals. Acupuncture reduced pain or discomfort,
promoted quality of life, or reduced drug-associated side-effects in cancer patients. Acupuncture also promoted the activities
of humoral and cellular immunity as well as natural killer (NK) cell activity. It improved patient survival and quality of
life as an adjunct with transcatheter arterial chemoembolization (TACE) for the treatment of hepatocellular carcinoma
and unresectable hepatocellular carcinoma, Chinese herbals improved patient survival time, quality of life, alleviation of
symptoms, and decreased marrow toxicity. Some herbs together with conventional protocols (chemotherapy or surgery)
demonstrated a promising outcome in lymphoma and breast cancer in both humans and dogs. The possible mechanisms
of herbal anti-cancer effects include: inhibition of the metabolism of tumor cells, alteration of tumor cell membranes and
information conduction, inhibition of tumor cell proliferation, induction of tumor cell apoptosis, enhancement of immune
function and anticancer capabilities, inhibition of neovascularization to prevent tumor growth and metastasis, prevention
of metastasis through improvement of the physical and chemical properties of blood cells, reversal of drug resistance and
enhancement of the effectiveness of chemotherapy.
Key words: Acupuncture, Chinese herbal medicine, traditional Chinese medicine, cancer, clinical application, oncology,
evidence based medicine, clinical trials
ABBREVIATIONS
MCT Mast cell tumor CHM Chinese herbal medicine

SCC Squamous cell carcinoma HCC Hepatocellular carcinoma
TCM Traditional Chinese medicine QOL Quality of life
WHO World Health Organization MST Mean survival time
CRT Chemoradiation therapy (CRT) PRST Pain-relieving sustained time
HNC Head and neck cancer
PEG Percutaneous endoscopic gastrostomy
Cancer is the most common natural cause of death in
BCRL Breast cancer-related lymphedema
dogs and cats in the United States. About 45% of dogs that
NK Natural killer
live to 10 years or older die of cancer.1-2 The most common
TACE Trans catheter arterial chemoembolization
cancers in pets are lymphoma, mammary tumors (benign/
UHCC Unresectable hepatocellular carcinoma
malignant), mast cell tumor (MCT), hemangiosarcoma,
soft tissue sarcomas, melanoma, squamous cell carcinoma
From: Department of Small Animal Clinical Sciences, College of
Veterinary Medicine, University of Florida, Gainesville, FL (Xie),
(SCC) and osteosarcoma.3-5 Traditional Chinese Medicine
Integrative Veterinary Oncology, 2501 N 32nd Street Phoenix, (TCM) has been used for the diagnosis and treatment of
AZ 85008 (Hershey), College of Traditional Chinese Veterinary tumors since the Warring States Period (475 to 221 BC),
Medicine, Agricultural University of Hebei, China (Ma)

when “Ling Shu Jing” (Miraculous Pivot) stated “the events and no infections or severe exacerbations after 255
tumor was caused by the pathological factors which had treatment sessions and 6 months of follow-up interviews.
stayed in the body over a long time”.6 Clinical evidence The authors concluded acupuncture for BCRL appears
indicates that acupuncture and Chinese herbal medicine safe and may reduce arm circumference.14
can be beneficial in the treatment of cancer in dogs and
people.7-13 This paper reviews acupuncture and Chinese Basic Research Studies on Immune Modulation
herbal medicine for the treatment or adjunct treatment of Associated with Acupuncture
cancers, and possible mechanisms of action. Five research studies that evaluated effects of
acupuncture on the immune system met inclusion criteria.
ACUPUNCTURE AND CANCER TREATMENT In the first study, Johnson et al reviewed 110 publications
Clinical Trials and concluded that acupuncture is associated with changes
There are 3 clinical trials that met inclusion criteria in natural killer (NK) cells, which secrete cytokines that
for this review that evaluated acupuncture treatment of inhibit or kill cancer cells.15 The use of acupuncture
cancer in human patients. In the first trial, cancer patients increased NK cell quantity and function in both humans
were divided into groups based on severity of pain (mild, and animals. It also could be used to promote the induction
moderate, severe) and then randomized into acupuncture and secretion of NK-cell activating cytokines that engage
group vs. medication group. The acupuncture group specific NK cell receptors that endogenously enhance
patients were treated with dry needling techniques at anticancer immune function. Three of the most commonly
3-5 severe A-shi or other related acupoints; for example, used acupoints in these studies were ST-36, PC-6 and LI-
BL-20, BL-21 and CV-12 were used for stomach cancer. 4. 15
Medication groups were treated with aspirin (mild The quantitative and qualitative effects of acupuncture
pain), codeine (moderate pain) and morphine (severe on the immune system was evaluated by collecting blood
pain) based on the World Health Organization (WHO) 3 from 17 healthy human volunteers aged 21-51 years old
step administration principle. Efficacy of pain relief of both before and after acupuncture treatment. The activity
acupuncture was 94.1% compared to the medicine group of T cells, B cells, macrophages and NK cells was
of 87.5% (P<0.05).7 In another randomized controlled assessed by using flow cytometry to measure CD-positive
clinical trial, it was shown that acupuncture treatment cell counts and cytokine expression levels. As compared
significantly promoted quality of life and reduced with the control group, a statistically significant increase
drug-associated side-effects of breast cancer patients in the number of CD2(+), CD4(+), CD8(+), CD11b(+),
medicated with anti-estrogens as compared with the sham CD16(+), CD19(+), CD56(+) cells as well as IL-4, IL-
acupuncture treatment.8 1beta and IFN-gamma levels in the cells were found in the
Dysphagia is a common side effect following acupuncture group after dry-needling stimulation at BL-
chemoradiation therapy (CRT) in patients with head 18, BL-20, BL-23 and ST-36. These observations indicate
and neck cancer (HNC). After dry needle at acupoints that acupuncture may regulate the immune system and
including ST-7, ST-36, LI-2, LI-11 and CV-23; a total promote the activities of humoral and cellular immunity
of 9 of 10 HNC patients (90%) post CRT reported an as well as NK cell activity.16
improvement in swallowing functions, xerostomia, pain, Tests were done on T cell subpopulations, NK
and fatigue levels. In addition, 6 of 7 (86%) percutaneous activity, humoral immunity and leukocyte count in
endoscopic gastrostomy (PEG) tube-dependent patients patients undergoing chemotherapy and acupuncture. Cell
had their feeding tubes removed after acupuncture, with levels were measured both before acupuncture treatment
a median duration of 114 days (range 49 to 368 days) and after 4 courses of electro-acupuncture was added at
post chemoradiation therapy versus PEG tube duration acupoints including ST-36, SP-6, CV-12 and PC- 6. Patients
(without acupuncture) with a median duration of 150 days undergoing combined acupuncture and chemotherapy
(range 178 to 365 days). 9 showed significantly higher leukocyte counts than that
In a study to evaluate the safety and potential efficacy of the control group (P < 0.01).17 The authors conclude
of acupuncture on upper-limb edema, 37 women with a electro-acupuncture may limit the immunologic damage
clinical diagnosis of breast cancer-related lymphedema associated with chemotherapy and can thus be used as
(BCRL) for 0.5-5 years were enrolled. The affected arm auxiliary therapy for patients undergoing chemotherapy.17
had a circumference ≥2 cm larger than the unaffected T lymphocyte subsets were also evaluated in another study
arm. Patients received dry needle acupuncture at TH-14, where human patients with malignant tumors were treated
LI-4, LI-15, LU-5, ST-36, SP-6, CV-3 and CV-12 twice in 30 minute sessions daily for 10 days with dry needle
weekly for 30 minutes over 4 consecutive weeks. The acupuncture at PC-6, LI-4, CV-4 + local acupoints. Results
mean reduction in arm circumference difference was of this study showed increases in T lymphocyte subsets,
0.90 cm (95% CI, 0.72-1.07; P < 0.0005). Eleven patients and beta-endorphin as well as decreased concentrations of
(33%) exhibited a reduction of ≥30% circumference after soluble interleukin 2 receptor.18
acupuncture treatment. There were no serious adverse

CHINESE HERBAL MEDICINE AND CANCER chemotherapeutic agents combining iodized oil chemo-
TREATMENT embolization and analgesics.22 The immediate and long-
Chinese Herbal Medicine for the Treatment of Liver term efficacy, adverse reaction, pain-relief initial time
Cancer-Clinical Trials (PRIT) and pain-relieving sustained time (PRST) of
A systematic review and meta-analysis of 37 the treatment, as well as the change in improvement of
randomized controlled clinical trials involving 2,653 patients’ quality of life (QOL) were observed. The CHM
patients was conducted to compare the efficacy and safety therapy group included using Oleum fructus bruceas
of Traditional Chinese Medicine (TCM) plus trans catheter (Ya Dan Zi You) intervention combined with oral intake
arterial chemoembolization (TACE) with that of TACE of Gan Ji Tang and external application of Ai Li Tong.
alone in treating unresectable hepatocellular carcinoma Gan Ji Tang ingredients included Codonopsis (Dang
(UHCC). The results showed that TCM+TACE compared Shen), Atractylodes (Bai Zhu), Bupleurum (Chai Hu),
with TACE alone, improved patient survival, quality of Paeonia (Chi Shao Yao), Curcuma (E Zhu), Eupolyphaga
life, alleviation of symptoms, and tumor response. No sinensis (Tu Bie Chong), Hedyotis Herb (Bai Hua She She
serious adverse events from TCM were reported and study Cao), Hirudo (Shui Zhi) and Ostrea (Mu Li). Ai Li Tong
results concluded the addition of TCM to TACE was contained Venenum Bfonis (Chan Su), Strychni (Ma Qian
therapeutically beneficial. 19 Zi), Cynanchi Paniculati (Xu Chang Qing), Moschus (She
In one retrospective study, 85 patients of 165 patients Xiang) and Borneolum syntheticum (Bing Pian). There was
with UHCC received TACE alone as the control group. no statistical difference (P>0.05) between the two groups
Another 80 patients (TACE + Chinese Herbal Medicine when disease control was evaluated, however, adverse
(CHM) group) received combined therapy consisting reaction occurrence rate was significantly less in the CHM
of TACE and a long-term oral herbal treatment with group when compared to the control group (P<0.05). The
Jiedu Formula granule (JDF). This CHM formula was pain-relief initial time (PRIT) was significantly superior
composed of 4 ingredients: root of Actinidia valvata (Mao in the herbal group when compared to the control group
Ren Shen), root of Salvia chinensis (Shi Jian Chuan), bulb (10.37±2.18 h vs 7.78±1.95 h, P<0.01). After treatment,
of Cremastra appendiculata (Du Juan Lan) and gizzard the CHM group patients’ somatic activity, symptoms
membrane of Gallus gallus domesticus (Ji Nei Jin). The and quality of life (QOL) were significantly improved
median overall survival (9.2 months) in the TACE+CHM when compared to those in the control group (P<0.05).
group was significantly longer than in the control group The survival rate in the two groups was similar at the 3rd
(5.87 months). The authors concluded that Chinese herbal month after treatment, but the CHM group was superior
medicine could improve the prognosis of patients with in terms of half- and 1-year survival rate (65.9% vs 42.5%
UHCC and prolong their survival.20 and 38.6% vs 18.1%, respectively, P<0.05). The median
In a case-controlled clinical trial, a total of 120 survival time in the CHM test group was 8.9 months and
patients with hepatocellular carcinoma (HCC) were that in the control group was 5.3 months.22, 23
randomized into two groups: transcatheter arterial Zhu reported that in a controlled study, 80 patients
chemoembolization (TACE) group (n=60) and TACE with primary middle and advanced stage liver cancer
+ CHM group (n=60). In TACE +CHM group, patients were randomly assigned to the TACE group (40 cases)
were given JDF and cinobufacini injection which was and to TACE plus herbal group (40 cases). Results
extracted from the skin of Bufo bufo gargarizans Cantor.21 showed that Kanglaite injection, extracted from herbal
There was a significant difference in survival rate between Coix seed oil (Yi Yi Ren), combined with TACE was
the two groups (P<0.01). The mean survival time (MST) significantly superior to TACE group alone in improving
of patients in the TACE+CHM group was 49.53 months symptoms and Karnofsky scores with effects such as
versus 39.90 months for the TACE alone group. The decreasing tumor growth and reducing marrow toxicity
1-, 2-, 3-, 4- and 5-year survival rates were 90%, 82%, for patients with advanced stage HCC.24
80%, 70% and 63%, respectively, in the TACE+CHM
group, and 79%, 70%, 60%, 60% and 36%, respectively, Chinese Herbal Medicine for the Treatment of
in the TACE alone group. A significant difference was Lymphoma-Clinical Trials
demonstrated in survival time between the two groups Lymphoma is the most common hematologic
(P=0.045) with Chinese herbal medicine able to postpone neoplasm in dogs. It is highly chemo-sensitive with
tumor recurrence and metastasis, prolong the survival complete remission rates ranging from 65 to 90% and
time and increase the survival rate of post-surgical patients median survival times of 26 to 51 weeks.25,26 There is
with HCC. 21 general consensus that combination protocols based on
In another prospective randomized controlled doxorubicin (e.g., in combination with cyclophosphamide,
design clinical trial, 97 patients with the middle/late stage vincristine, and prednisone--CHOP) are associated with
of HCC were assigned to the CHM group (49 cases) the longest disease-free interval and survival times.26
treated with Chinese herbal medicine therapy alone and Reported side effects of the CHOP protocol, however,
the control group (48 cases) which were treated with include sterile hemorrhagic cystitis, myelosuppression,

and gastrointestinal toxicity.26 While the treatment for Basic Research Studies on Development of New
lymphoma is efficacious with well-established outcomes Anticancer Chinese Herbal Medicines and Lymphoma
(survival time, time to complete or partial remission, and Wogonin (5,7-dihydroxy-8-methoxyflavone), isolated
disease-free interval), clients and oncologists equally from the roots of the traditional Chinese herb Scutellaria
emphasize the importance of a good quality of life (QOL) baicalensis Georgi (Huang Qin), has been recently
in cancer bearing dogs when selecting treatment options.27 recognized as a new anticancer drug that possesses
The Acupuncture Service at University of Florida a cytotoxic effect against a large panel of human
Small Animal Hospital treated 10 canine patients with cancer cell lines by inducing apoptosis in vitro. It was
lymphoma from 1999 to 2012. The final outcome was demonstrated that the application of drug-coated magnetic
determined in 7 of the 10 cases with 3 cases undetermined nanoparticles (MNPs) could strengthen wogonin-induced
due to loss of contact with the client. The overall QOL cell inhibition, apoptosis, and cell cycle arrest in Raji
including activity level, mental stimulation, social cells. This was documented by methylthiazol tetrazolium
relations, appetite and stool were generally better after assay, flow cytometer assay, and nuclear 4′,6-diamidino-
acupuncture and herbal medicine treatment in all 7 cases 2-phenylindole staining. The molecular mechanisms of
with a complete medical record. A combination of CHOP these phenomena were further explored by western blot, in
and acupuncture/Chinese herbal medicine was used in 5 which the protein levels of caspase 8 and caspase 3 were
cases while acupuncture/herbal medicine alone was used increased significantly while those of survivin and cyclin
in 2 cases. The survival time of 5 lymphoma dogs which E were decreased significantly in wogonin-MNPs group.
received both chemotherapy and acupuncture/herbal These findings suggest that the combination of wogonin
medicine ranged from 8 to 56 months (average of 25.2 and MNPs provides a promising strategy for lymphoma
months) while the survival time of 2 dogs receiving only therapy.30
acupuncture/herbal was 7 and 8 months. The three primary
Chinese herbal medicines used in these cases were Wei Chinese Herbal Medicine for the Treatment of
Qi Boostera (Modified Si Jun Zi Tang), Stasis Breakera Mammary Tumors
(Modified Nei Xiao Wan) and Max’s Formulaa (Modified Ninety-six Chinese herbal medical formulae have
Nei Xiao Luo Li San). The ingredients and indications for been used for the treatment of breast cancer (1990 - 2010)
these Chinese herbal medications have been published.28 in China with the most commonly used herbs including
Twenty-seven patients with malignant lymphoma (14 Astragali (Huang Qi), Atractylodis Macrocephalae (Bai
Hodgkin’s disease, 9 histiocytosarcoma, 2 plasmacytoma, Zhu), Poriae Cocos (Fu Ling), Pseudobulbus Shancigu
2 lympholeucosarcoma) were treated with a combination (Shan Ci Gu), Angelicae Sinensis (Dang Gui), Coix (Yi Yi
of western medicine (Ailin-injection) and Chinese herbal Ren), Herba Hedyotidis Diffusae (Bai Hua She She Cao),
medicine. The latter’s composition and dose varied and Curcuma Phaeocaulis (E Zhu).31 Other herbals for the
depending on the clinical manifestations. All patients treatment of breast cancer included Rabdosia rubescens
had stage III and IV lesions. The total remission rate was (Dong Ling Cao), Sanguisorba officinalis L (Di Yu) and
70.37% (complete remission 13 patients, partial remission Cornus officinalis (Shan Zhu Yu).32-33
6 patients, no remission 8 patients). In addition, in cell Eight dogs with histologically confirmed mammary
line studies, the suppressive effect of Ailin on transplanted adenocarcinoma had surgical excision of their cancer
tumor in vivo (S-180) was studied. Three repetitions of the and no evidence of metastasis on thoracic radiographs.
treatment yielded suppression rates of 36.09%, 48.80%, All dogs were administered 0.022 mg/kg Mammosolb,
32.67% with an average of 39.18% (P < 0.002). The same a proprietary blend of 28 herbal ingredients, mainly
treatment using S37 cells and Ehrlich ascites tumor cells including Bubali (Niu Jiao), Olibanum (Ru Xiang), Myrrh
yielded suppression rates of 37.25%, 35.29% (average (Mo Yao), Astragalus (Huang Qi), etc., twice daily for
of 36.27%; P < 0.005) and 46.53%, 44.56%, 51.35%, 2 years and monitored with thoracic radiographs and
45.95% (average of 47.09% ; P < 0.001), respectively. The physical examinations at 3, 6, 9, 12, 18 and 24 months
clinical and experimental studies suggest that Ailin-1 had post-surgery. Serum biochemical tests for liver and kidney
no serious side effects and was well tolerated by patients function were evaluated during the study period to monitor
with malignant lymphoma at moderate or advanced for unexpected toxicity. In this study, 7 of the 8 dogs lived
stages with no toxic reactions observed at clinical dose. greater than 2 years. The dogs that lived less than 2 years
In the above combination therapy (Ailin plus CHM), the only received half the dose of herbs due to improper
addition of the Chinese herbal medicine had a beneficial caretaker compliance. Survival rates were significantly
supplementary effect on the treatment of patients’ cachexia increased in the dogs that had surgical excision and
and immuno-suppression along with improved remission Mammosol as compared to previously published results
rates in humans (70%) versus tumor suppression in cells of dogs that had surgery alone (P<0.001) and dogs with
lines with Ailin only (36.27% and 47.09%).29 surgery and chemotherapy (P<0.05) using the discrete
Bonferroni method.34

Chinese Herbal Medicine for Treatment of Melanoma an indirect anti-neoplastic effect. 41

and Acanthomatous Epulis
A combination of Wei Qi Boostera (Modified Si Jun Alteration of tumor cell membranes and information
Zi Tang), Stasis Breakera (Modified Nei Xiao Wan) and conduction
Max’s Formulaa (Modified Nei Xiao Luo Li San) was In a controlled study, 48 mice were randomly
used for the treatment of an equine case of unresectable assigned to 3 groups, untreated control group,
melanoma in a horse and a canine case of acanthomatous experimental model group and model with herbal group.
epulis along with acupoints including BL-20, BL-21, Results showed the herbal mixture of Rehmannia (Sheng
LI-4, ST-36 and ST-40.35-36 The unresectable amelanotic Di), Isatis (Ban Lan Gen), and Oldenlandia (Bai Hua She
melanoma mass (11 x 7 x 4 cm3), located on the ramus of She Cao) can significantly reduce the level of cGMP in
the right mandible in a 7-year-old intact female Arabian a HepA hepatoma mouse model and increase the cAMP/
horse, completely resolved 4 months after treatment with cGMP ratio.42 This study suggested that the anti-tumor
Chinese herbal medicine and acupuncture treatments.35 It mechanism of these herbs is related to a change in the
has remained in remission for 9 yrs, when the paper on this mobility (fluidity) of the cell membrane and regulation of
case was published, at the age of 16 years old. A maxillary information along the cell signal conduction pathway of
acanthomatous epulis mass was present in a 5-year-old tumor cells.
male Chihuahua. It was located on the right side of the
maxilla over the incisors and canine tooth with a median Inhibition of tumor cell proliferation
diameter of 2.4 cm. Three months after therapy with The inhibition of tumor cell proliferation is another
Chinese herbal medicine and acupuncture, the mass began important area of onco-molecular biological research.
to shrink and was almost completely resolved 12 months A randomized controlled study by Qin SY et al.
after the start of treatment.36 It remained in remission for demonstrated that extracts of Chinese fan palm seed (Pu
6 years, at which time the dog died of an unrelated event Kui Zi) and Pyrola (Lu Ti Cao) had a clear dose-dependent
(congestive heart failure). Preliminary data on the basic and significant anti-tumor activity on a HeLa cell line
composition of these Chinese herbal products have been (cells derived from human cervical carcinoma), with IC50
recently published.37 (95.51 and 95.40 mg·L-1), respectively. Chinese fan palm
Based on anticancer in-vivo and in-vitro studies seed (Pu Kui Zi) has also been shown to have an inhibitory
(inducing apoptosis in cancer cells), individual Chinese effect on Bel-7402 tumor cell lines, with IC50 (122.84
herbs that showed promise included Sparganii Stoloniferi mg·L-1).43
(San Leng),38 Curcuma phaeocaulis (E Zhu)39 and Panax
notoginseng (San Qi) extracts.40 Both Sparganii Stoloniferi Induction of tumor cell apoptosis
(San Leng) and Curcuma phaeocaulis (E Zhu) could purge The apoptotic effects on tumor cells of matrine and
the interior, break Blood stasis and clear masses. These oxymatrine, two major alkaloid components extracted
Chinese herbs have been widely used for any tumor in mainly from Sophora japonica (Ku Shen), Sophora
traditional Chinese medicine. Their anti-neoplastic effects subprostrata (Shan Dou Gen) and Sophora alopecuroides
also have been confirmed by modern pharmacological (Ku Gan Cao), have been studied by Si KW et al.44 Matrine
experiments and clinical studies.38,39 Panax notoginseng induced apoptosis of HepG2 cells (a human hepatic
(San Qi) can stop hemorrhage and resolve Stagnation, carcinoma cell line) was demonstrated by morphologic
exerting beneficial effects on any hemorrhage and changes, increased DNA breakdown using a TUNEL assay
stasis conditions. One study demonstrated that Panax in-situ, observing apoptosis peaks with flow cytometry
notoginseng extracts had significant apoptosis-promoting and detection of early stage apoptosis using an AnnexinV-
effect on MNNG-transformed GES-1 cells.40 FITC/PI double labeled assay.45 The mechanism of action
of matrine to induce HepG2 cell apoptosis was to up-
RESEARCH STUDIES ON MECHANISM OF regulate the expression of apoptotic genes wp53 and box
ACTION OF CHINESE HERBAL MEDICINE ANTI- and down-regulate the expression of anti-apoptotic genes
CANCER EFFECTS bcl-2, which eventually led to cell apoptosis.
Inhibition of the metabolism of tumor cells
Rhubarb (Da Huang) affects numerous elements Enhancement of immune function and anticancer
of cancer cell metabolism and can cause inhibition of capabilities
respiration, oxidation and dehydrogenation of amino acids Most tumor cells are destroyed by host NK cells,
and glucose metabolism. It can also decrease DNA, RNA macrophages and neutrophils. In previous studies only
and protein biosynthesis of tumor cells and inhibit tumor 1% of tumor cells survived after 24 hours when injected
cell proliferation with no significant impact on normal intravenously into animals and 0.1% of cells could form
cells. Rhubarb (Da Huang) can also significantly improve pulmonary metastasis.45 There are many opportunities for
the efficacy of cellular immunity in mice and can promote tumor cells to be destroyed in the blood stream. Traditional
lymphocyte proliferation and IL-2 synthesis, which play Chinese medicines enhance immune function and

significantly improve the activity of the natural killer (NK) Prevention of metastasis through improvement of the
cells, macrophages and neutrophils which remove tumor physical and chemical properties of blood cells
cells. PSK (Kreston) is a protein-bound polysaccharide Blood viscosity is closely related to tumor metastasis
obtained from cultured mycelia of Coriolus versicolor and there have been studies that have shown that increased
basidiomycetes mushrooms (Yun Zhi) and is a biological blood viscosity can promote tumor metastasis and
response modifier with diverse actions, one of which adhesion of tumor cells under certain conditions.51-53 In
includes anticancer properties. PSK has been shown to the invasive stage of tumors, plasma viscosity increases
suppress pulmonary metastasis of methylcholanthrene and microthrombi containing cancer cells form. Adhesion
induced sarcomas, human prostate cancer cells (DU145M between cancer cell and capillary endothelium develops in
cell line) lymphatic metastasis of mouse leukemia cancer these hypercoagulative states. The Chinese herbal formula
cells (P388 cell line) and prolong the survival period in Huo Xue Hua Yu and traditional Chinese medicines
spontaneous metastasis models.46 The effects of oral PSK Salvia (Dan Shen), Leonurus (Yi Mu Cao), Rhubarb (Da
were also examined in CDF1 mice with liver metastases of Huang), Paeonia (Chi Shao), Carthamus (Hong Hua),
Colon 26 adenocarcinoma. The survival of tumor-bearing Curcuma (Jiang Huang), Sappan (Su Mu), Leech (Shui
mice was prolonged and both metastatic foci and liver Zhi), Ligusticum (Chuan Xiong), Sparganium (San Leng),
weight were decreased. E Zhu (Zedoaria), Blister bug (Ban Mao), Polygonum (Hu
Rhubarb (Da Huang) and Rehmannia 6 (Liu Wei Zhang) and others activate blood circulation and dissipate
Di Huang Tang) can enhance proliferation of spleen Blood Stasis to reduce hypercoagulation of the blood and
T-lymphocytes and secretion of interleukin-2 in S180 tumor metastasis.54 A study by Cui W et al. showed that
tumor-bearing mice causing an immune-mediated anti- the Chinese herbal formula Huo Xue Hua Yu (Activating
tumor effect.47 The Chinese herbal formula Jian Pi Yi Blood Circulation to Dissipate Blood Stasis) and one of
Shen preparation for infusion, composed of Codonopsis its herbal ingredients Millettia (Ji Xue Teng) significantly
(Dang Shen), Atractylodes (Bai Zhu), Lycium (Gou Qi inhibited platelet aggregation of tumor cells (P<0.001),
Zi), Ligustrum (Nv Zhen Zi) and others, can increase the suggesting these as another potential Chinese medicine to
capacity of macrophage phagocytosis and the overall inhibit hematogenous tumor metastasis.55
number of T lymphocytes in the peripheral blood and
inhibit Lewis lung cancer metastasis and U14 lymphatic Reversal of drug resistance and enhancement of the
metastasis via the blood stream.48 One review paper effectiveness of chemotherapy
summarized the status of the anti-leukemia effects of One of the main reasons that conventional
Traditional Chinese Medicine and naturally occurring chemotherapy fails is because multi-drug resistance
drugs. It stated that Polysaccharides from Achyranthes (MDR) develops as the tumor cells become tolerant to
(Niu Xi), Rehmannia (Di Huang) and Poria (Fu Ling) various agents. P-glycoprotein (P-gp) is a polymorphic
have been shown to enhance immune function. The efflux transporter located on cell membranes. In human
Chinese herbs Ginseng (Ren Shen), Astragalus (Huang tumors, the MDR phenotype is associated with an
Qi), Codonopsis (Dang Shen), Atractylodes (Bai Zhu), over-expression of the P-gp encoded by the multi-drug
Angelica (Dang Gui), Salvia (Dan Shen), Zedoaria (E resistance 1 (MDR1) gene and an increase in the ATP-
Zhu) and others strengthen the immune system as well.49 binding membrane protein 190 kDa encoded by a MRP
gene (multidrug resistance-associated protein (MRP)),
Inhibition of neovascularization to prevent tumor which results in lower intracellular drug concentrations.
growth and metastasis When MDR occurs there is an overproduction of Pgp,
Tumor cells obtain nutritional support and send an increase in the activity of the glutathione (GSH)
malignant cells out to the host through blood vessels, detoxication enzyme system and alteration of the DNA
which results in enhanced tumor growth and metastasis topoisomerase enzyme activity which improves DNA
respectively. An important antitumor strategy is to inhibit repair.56
neovascularization in order to “shut off the lifeline” of The effects of andrographolide extracted, from
the tumor cells. The search for effective angiogenesis Andrographis (Chuan Xin Lian), to reduce the
inhibiting factors (AIF) has evoked a strong interest in development of MDR was studied by Han Y et al.57
traditional Chinese medicines. Some traditional Chinese Different concentrations of andrographolide reversed the
medicines like Ten Large Tonification (Shi Quan Da Bu MDR effect of large intestine carcinoma cells (HCT-8/5-
Tang), Minor Bupleurum Decoction (Xiao Chai Hu Tang), FU cell line) as determined by a standard MTT assay.57
Coriolan (Yun Zhi Duo Tang), Lentinan (Xiang Gu Duo From their studies, they concluded that andrographolide
Tang), Angelica (Dang Gui), Sichuan peony (Chuan could not only increase the tumor toxicity of chemotherapy
Shao), Cinnamomum (Gui Zhi) or extracted components drugs, but to some degree reverse drug-resistantance of
like Cordyceps (Dong Chong Xia Cao) polysaccharide cell lines. The Chinese herbal formula Fu Fang San Gen
and others can induce tumor necrosis factor (TNF).49-50 is composed of Polygoni cuspidate root (Hu Zhang Gen),
Yangtao Actinidia root (Teng Li Gen) and Thin leaf adina

root (Shui Yang Mei Gen). Xie CS et al. used an MTT incidence rate and distribution of tumours in 4,653
assay to determine the ability of the Fu Fang San Gen cases of archival submissions derived from the
to reverse MDR effects in two multi-drug resistant cell Dutch golden retriever population. BMC Vet Res.
lines, K562/ADR and K562/VC (2 types of MDR human 2014; 10:34. doi: 10.1186/1746-6148-10-34.
erythroleukemia cells).58 Fu Fang San Gen partially 4. Merlo DF, Rossi L, Pellegrino C, et al. Cancer
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of telomerase activity and reversal of drug resistance to of traditional Chinese medicine and acupuncture
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Highway Speonk, NY 15. Johnston MF, Ortix Sanches E, Vujanovic NL, et
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Chinese) PET | TAO was created to provide solutions for supporting
53. Ma WJ, Cheng GZ. The relationship between and nurturing harmony in the lives of animals.
hematogenous metastasis and blood viscosity of
tumors. Intermediate Medical Journal. 1992; 27(4):8. We do this by offering nutritious products that combine
(In Chinese) Eastern medical principles with Western nutritional science.
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54. Huang J, Ma LH, Liu JX, et al. The application of
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Traditional Chinese drugs for Activating Blood
circulation to dissipate Blood Stasis and tumor www.pettao.com info@pettao.com
metastasis. Forum on Traditional Chinese Medicine.
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2001; 16 (4):11-12. (In Chinese)

t e r n a t io n a l
In
19th Annual ference 2017
TCVM Con bilitation | Reha
- of-li fe Ca r e | Geriatric
End
ada
Toronto, Can
, 2017
Conference Theme Traditional Chinese Veterinary Medicine
August 18-20
• TCVM for End-of-life Care: TCVM for Hospice and Palliative Care, tips to communicate with caregivers, how
to support caregivers and veterinarians, how to relieve distress;
• TCVM for geriatric conditions: Including cognitive dysfunction, cancer, osteoarthritis, renal failure, heart
failure, liver failure, Wei syndrome (rear weakness);
• TCVM for pediatric conditions: Including Orthopedic, Neurological, Gastrointestinal Immunological diseases,
special food therapy and Tui-na for Pediatric Disharmonies;
• Rehabilitation to enhance acupuncture results.
Main Speakers
Huisheng Xie Bruce Ferguson Tami Shearer Hanwen Cheng

DVM, PhD DVM, MS, CTCVMP DVM, CVA DVM, MS, CVA
The 19th Annual International TCVM Conference will be held in Toronto, Canada from August 18 to 20, 2017.
This will be the first ever Annual Conference to take place in Canada. This International TCVM Conference
is co-sponsored by the World Association of TCVM and the Chi Institute. The Annual TCVM Conference
provides a great opportunity for veterinarians to learn from the top TCVM experts, meet TCVM practitioners
from all over the world.
Registration Fee Location Co-sponsored by

• Early Bird Registration The conference will take place at the World
(register by June 1, 2017): $795 Westin Prince Toronto.
• Advanced Registration
Association
Address: 900 York Mills Road
(register by July 18, 2017) $895 Toronto, ON M3B 3H2 Canada
of TCVM
• Regular Registration Web: westin.com/prince
(register after July 18,2017): $975 To register or for more information
Phone: 1 416-444-2511
• Reception Banquet: $70 contact the Chi Institute:
All phone-in reservations must be made to Visit www.tcvm.com
Registration fee is due upon registration 416-444-2511 or 1-888-627-8550, by July
and covers: Breakfast and lunch during 17, 2017. Please refer to the “Chi Institute”
Call 800-860-1543
conference, conference proceedings and to receive the group rate of $139 CAD/ E-mail register@tcvm.com
handout, parking and WiFi access. night.

Brief Communications
Preliminary Evaluation of the Effect of

Acupuncture on Acoustic Myographic
Recordings in Five Sport Horses
Esther M. Millares DVM, Helga Bollingberg-Soerensen,

Sarah S. Le Jeune DVM, DACVS, DACVSMR
ABSTRACT
The objective of this prospective controlled study was to assess the effect of acupuncture (ACU) on acoustic myography (AM)
of the middle gluteal muscle at the walk and trot in healthy sport horses (n=5). Acoustic myographic sensors were placed on
the surface of the right and left middle gluteal muscles. Recordings were taken while the horses were walked and trotted in
hand, in a straight line, before and immediately after ACU treatments, and 48 hours and one week afterwards. Acoustic myo-
graphic data was used to calculate an acoustic myographic (EST) score. Due to the small number of horses in this pilot study,
data was presented descriptively. Gait abnormalities and sensitivity at routine ACU points were recorded at each session.
All horses tolerated the procedure well and did not exhibit any signs of lameness at any time. Sensitivity at ACU points was
present initially in 3 horses and was abolished immediately after ACU in all of them. All horses had an increased EST score
after 48 hours, of which 3 had a persistent increase for 1 week afterwards, suggesting a greater speed of activation of muscle
fibers after ACU. AM is a non-invasive technique, which is well tolerated and easily performed in a routine clinical setting.
This small pilot study suggests a change in acoustic myographic data after ACU treatment but this will need to be confirmed
in a greater number of horses.
Key words: Acupuncture, acoustic myography, sport horses
shown that needle stimulation of acupuncture points

ABBREVIATIONS
causes microtrauma that produces a local inflammatory
reaction, which results in an increased local tissue immune
ACU Acupuncture
response, improved local tissue blood flow, and muscle
AM Acoustic myography
relaxation.1-6 Studies in humans, have suggested that ACU
DAPE Diagnostic acupuncture palpation exam
might stimulate motor nerve fibers, and as such, increase
E Efficiency
muscle activity.7, 8 Other studies suggest that the effect of
S Spatial amplitude
ACU might be related to the inhibition of excitability of
T Temporal summation
the alpha motor neuron. 9
EST score Acoustic myographic score
In horses, the use of ACU as a therapeutic modality
has increased significantly over the last 20 years, however
Acupuncture (ACU) is part of Traditional Chinese scientific evidence to support this practice is somewhat
Medicine and involves stimulating points on the body lacking.10 Recently, multiple studies have been performed
surface with acupuncture needles. The effects of ACU to assess the efficacy of ACU in the treatment of various
cannot be explained in terms of a single mechanism, conditions, such as Equine Herpes Virus type 1, chronic
but involve a series of interactions between the nervous, back pain and laryngeal hemiplegia. 5, 11-13 Additionally,
endocrine, and the immune systems. Mechanisms of sport horses that have sensitivity to palpation of ACU
action are still not fully understood but studies have points are more likely to be lame.14 There continues to be
a need for clinical studies proving the efficacy of ACU.
From: William R. Pritchard Veterinary Medical Teaching Hospital Acoustic myography (AM) is a non-invasive
(Millares) and the Department of Surgical and Radiological
Sciences (le Jeune), School of Veterinary Medicine, University
technique that measures muscle activity by capturing the
of California-Davis, Davis, CA, USA and the Faculty of Health sounds generated by muscle contractions.15-17 The CURO
and Medical Sciences (Bollingberg-Soerensen), Copenhagen system®a is a commercially available AM system that has
University, Frederiksberg, Denmark

been validated in horses.18 The objective of this study was month of March at the same time and in a consistent order.
to assess the effect of ACU treatment on AM recordings The AM measurements and diagnostic acupuncture
of the middle gluteal muscle in healthy sport horses. We palpation exam (DAPE) were recorded before and after
hypothesized that ACU, would improve the efficiency of ACU, on day 0, 2, and 7, at the walk and trot. The DAPE
muscle contraction during exercise, and this would be was performed as previously described immediately prior
reflected on the AM recordings. to ACU.14 ACU was performed by the same certified
equine veterinary acupuncturist (S.L.J.) and consisted
MATERIALS AND METHODS of placement of sterile, stainless steel, 0.30 mm x 50
Five client-owned sport horses (Warmblood geldings) mm AP needlesb at fifteen standard and commonly used
were included in the study after approval by the Animal acupoints: PC-9 bilaterally, Bai-Hui, GB-21 bilaterally,
Care and Use Committee of the University of California, BL-10 bilaterally, BL-21 bilaterally, BL-23 bilaterally,
Davis. All horses were deemed healthy based on physical BL-54 bilaterally, BL-40 bilaterally.19, 20 The needles
and lameness examinations and the absence of recent were inserted to a depth of 20 mm, no manipulation of
medical issues. These horses were ridden regularly and the needles was performed after insertion, and they were
competing in showjumping and were free of any forelimb left in place for ten minutes after the last needle was
or hindlimb gait abnormality. placed. On each treatment day, sensitivity to the DAPE
Acoustic myographic (AM) data was recorded from was evaluated and recorded prior to the AM recording as
each horse at the walk and trot in a straight line. The previously described (Table 1).14
horses were evaluated at their respective training facility AM recordings were transformed into E, S, T values
on a consistent surface, either a sand- or gravel-based hard by a commercially available acoustic myographic systema.
surface. The horses were handled by the same handler This system measures 3 parameters involving muscular
throughout the study period. Each horse was instrumented contractions: efficiency (E),which is the way the Central
with a commercially available system (CURO system®)1 Nervous System (CNS) recruits and uses the active fibers
consisting of 2 single-use calibrated sensors (dimensions in a muscle, spatial amplitude (S), or the number of
9.5x14.5x3.5cm; weight 290g) (Figure 1). Each sensor active fibers and temporal summation (T), defined as the
was placed 3 inches abaxially to the pelvic midline over frequency with which the muscular fibers contract. The
the middle gluteal muscle, after the skin was cleaned acoustic myographic score or EST score was calculated by
with water and prepared with ultrasound gel to increase the following formula: EST average left: (EL+SL+TL)/3
the contact between the sensor and the skin. The sensors and EST average right: (ER+SR+TR)/3.
were kept in place with adhesive foam tape included with Statistical evaluation of data in this pilot study was
the CURO systema (Figure 2). The CURO systema was not performed due to small study animal size which does
placed in a pouch and attached to a surcingle, for real time not allow meaningful statistical interpretation. Study data
recordings. Data were digitally sampled at 2-250 Hz in is therefore presented descriptively.
real time and analyzed by the CURO softwarea. Horses
were evaluated on day 0, day 2, and day 7 during the
The CURO system
Weight: 290 g
Width 14.5 cm
Breadth: 3.5 cm
12 V dc charger + USB link
4 muscles (2 antagonist/agonist
groups)
Real-time assesment
ESTITM Analysis & Balance Score
Recording time: 7 hours non-stop
Figure 1: Image of the CURO system including Figure 2: Image of a horse instrumented with the
specifications. Myodynamik Aps (Denmark) CURO system

RESULTS Of the 5 horses included in the study, 3 showed

The 5 horses included in this study were client- sensitivity to DAPE initially. None of the horses showed
owned Warmblood geldings, ranging in age from 6 to 15 sensitivity to the DAPE 48 hours after ACU. One week
years (mean age 9 years, median 6 years), and were all after ACU treatment, 4/5 horses showed no sensitivity to
professionally trained show-jumping horses. The horses the DAPE scan and 2/5 horses had no sensitivity to DAPE
were deemed systemically healthy, and no evidence of at any time during the study (Table 2). All horses had an
lameness was observed at any time point during the study increased EST score after 48 hours at the walk and trot,
period. All horses tolerated the surcingle with the AM with 3 horses demonstrating a persistent increase for 1
system and the ACU treatment well. week following ACU treatment (Figure 3 and 4).
Table 1: Criteria for acupoint sensitivity during DAPE. Le Jeune SS et al. Prospective Study on the Correlation of Positive
Acupuncture Scans and Lameness in 102 Performance Horses AJTCVM Vol 9, No.2, August 2014
Grade Criteria
1 An inconsistent localized flinch
2 A mild consistent localized flinch
3 A moderate consistent flinch
4 A severe flinch
5 Severe flinching with evasion and may kick or bite
Table 2: Comparison of the sensitivity to the DAPE in each individual horse along the study period time. Horses are listed in the
order the procedure was performed. ( ) grade of sensitivity according to ‘criteria’ previously described: le Jeune SS et al, 2014
Day 0 Day 0 Day 2 Day 2 Day 7 Day 7
Horse
Left side Right side Left side Right side Left side Right side
Stomach 10 (2)
Bladder 18-19 (3)
#1 Bladder 18-19 (3) Clear Clear Bladder 18-21 (2) Clear
Bladder 54 (4)
Bladder 54 (4)
#2 Bladder 54 (2) Clear Clear Clear Clear Clear
Bladder 23 (2)
#3 Clear Clear Clear Clear Clear
Bladder 54 (1)
#4 Clear Clear Clear Clear Clear Clear
#5 Clear Clear Clear Clear Clear Clear
Figure 3: Comparison of the mean EST scores of the left and right middle gluteal muscles while horses were walked in a straight
line, over the course of the study. This graph represents the beneficial effects observed in 3/5 horses in the present study.
DISCUSSION muscle.15-17 The main advantage of AM is therefore that

There is a need for evidence based clinical studies the innervation zone or the tendinous regions can be used
evaluating the efficacy of acupuncture, particularly for for sensor placement. Thus the AM sensors can be placed
lameness in horses. Sensitivity at certain acupoints has anywhere on the muscle of interest to receive a reliable
previously been correlated to pain at specific anatomic signal.
sites in one study which demonstrated the association of a The acupuncture palpation exam (DAPE) was
positive DAPE with lameness in sport horses.14, 21-23 Results performed by the same certified acupuncturist (SLJ), just
of that study support that positive responses to the DAPE prior to AM measurement, reducing variability in this
are closely associated with lameness (accuracy, sensitivity measurement in the study. The AM measurements (EST
and specificity near 80%). Acoustic myography (AM) is a score) and DAPE measurements were recorded both before
non-invasive technique that measures muscle activity by and after ACU in sound horses on a straight line at walk
capturing the sounds generated by muscle contractions. and trot. Encouragingly, 5/5 horses had increased EST
It appears to lend itself to measuring changes in muscle scores 48 hours after ACU, suggesting an improvement
use which can be correlated with pain detection by DAPE of the muscle efficiency after ACU which supported the
and pain relief associated with ACU treatment.15-17 The use study hypothesis. In addition, there was improvement in
of AM has been validated both in research horses in the DAPE scores (5/5) at 48 hours post ACU and persisted in
gluteus medius muscle and in humans demonstrating its 3 horses at 1 week.
robust potential for assessment of force production and The results of this study suggest that ACU could
fatigue.17, 18, 24 This procedure is particularly appealing for affect muscle by altering the efficiency of the contractions
use in performance horses as it is non-invasive and could which is important to equine locomotion. These results
easily be performed in a clinical setting on any superficial are tempered by the limitations of this pilot study which
muscle. are small sample size, no control group, no lameness in
The objective of this pilot study was to assess whether the study group and horses evaluated in a straight line so
or not ACU has an appreciable effect on AM recordings, subtle gait asymmetry might not be detected. Even with
as measured by the effect on muscular activity of the these limitations, this study demonstrated that AM is a
gluteus medius in sport horses. This muscle was selected non-invasive technique which is well tolerated and easily
because of its previous validation for AM use, superficial performed in a routine clinical setting. The effect of ACU
location and its importance for propulsion as extensor of on AM was easily identified by changes in AM data after
the hip and abductor of the limb.25 The position of the AM ACU treatment in the study horses. The results of this pilot
sensors was selected as to not interfere with the DAPE and study are encouraging, therefore, further studies need to
ACU. Unlike electromyography, which is very placement be performed in a greater number of horses along with
sensitive in terms of signal strength, the AM signal remains inclusion of lame horses to evaluate the effects identified
relatively constant throughout the surface of a contracting in this study and validate these statistically.
Figure 4: Comparison of the mean EST scores of the left and right middle gluteal muscles while horses were trotted in a
straight line, over the course of the study. This graph represents the beneficial effects observed in 3/5 horses in the present
study.

record. 2009;165(20):602-4.
FOOTNOTES 13. Klide AM, Martin BB, Jr. Methods of stimulating
a. CURO System, MyoDynamiks Aps, Denmark acupuncture points for treatment of chronic back
b. Vinco, Helio Medical Supplies, Inc, San Jose, CA pain in horses. Journal of the American Veterinary
Medical Association. 1989;195(10):1375-9.
14. Le Jeune SS. Prospective Study on the Correlation
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KO, Fairhurst H, Jackson K, et al. A pilot study of 15. Barry DT, Geiringer SR, Ball RD. Acoustic
the effects of acupuncture treatment on objective and myography: a noninvasive monitor of motor unit
subjective gait parameters in horses. Vet Anaesth fatigue. Muscle & nerve. 1985;8(3):189-94.
Analg. 2016. 16. Harrison AP, Danneskiold-Samsoe B, Bartels
2. Kawakita K, Okada K. Acupuncture therapy: EM. Portable acoustic myography - a realistic
mechanism of action, efficacy, and safety: a noninvasive method for assessment of muscle
potential intervention for psychogenic disorders? activity and coordination in human subjects in most
BioPsychoSocial medicine. 2014;8(1):4. home and sports settings. Physiological reports.
3. P B. Acupuncture, Trigger Points and Musculoskeletal 2013;1(2):e00029.
Pain. 3rd Edition ed: Elsevier; 2005. 17. Smith TG, Stokes MJ. Technical aspects of acoustic
4. Shmalberg J, Xie H. Acupuncture and Chinese herbal myography (AMG) of human skeletal muscle:
medicine for treating horses. Compendium (Yardley, contact pressure and force/AMG relationships.
PA). 2011;33(5):E1-11. Journal of neuroscience methods. 1993;47(1-2):85-
5. Xie H, Colahan P, Ott EA. Evaluation of 92.
electroacupuncture treatment of horses with signs of 18. Riis KH, Harrison AP, Riis-Olesen K. Non-invasive
chronic thoracolumbar pain. Journal of the American assessment of equine muscular function: A case
Veterinary Medical Association. 2005;227(2):281-6. study. Open veterinary journal. 2013;3(2):80-4.
6. Chan WW, Chen KY, Liu H, Wu LS, Lin JH. 19. Fleming P. Equine Atlas of Acupuncture Loci. 2nd
Acupuncture for general veterinary practice. The edition ed. Veterinary Acupuncture: Ancient Art to
Journal of veterinary medical science / the Japanese Modern Medicine2000.
Society of Veterinary Science. 2001;63(10):1057-62. 20. Xie H, Trevisanello L. Equine Transpositional
7. Chou LW, Hsieh YL, Kao MJ, Hong CZ. Remote Acupoints. Xie’s Veterinary Acupuncture: Blackwell
influences of acupuncture on the pain intensity Publishing Ltd; 2008. p. 27-87.
and the amplitude changes of endplate noise in 21. Alfaro A. Correlation of Acupuncture Point
the myofascial trigger point of the upper trapezius Sensitivity and Lesion Location in 259 Horses.
muscle. Archives of physical medicine and AJTCVM. 2014;9(1):83-7.
rehabilitation. 2009;90(6):905-12. 22. McCormick W. The origins of acupuncture channel
8. Dar G, Hicks GE. The immediate effect of dry imbalance in pain of the equine hindlimb. Journal of
needling on multifidus muscles’ function in healthy Equine Veterinary Science. 1998;18.
individuals. Journal of back and musculoskeletal 23. AM S. Equine Acupuncture for Lameness Diagnosis
rehabilitation. 2015. and Treatment. Diagnosis and Management of
9. Sandberg M, Lindberg LG, Gerdle B. Peripheral Lameness in the Horse 2003.
effects of needle stimulation (acupuncture) on skin 24. Stokes MJ, Dalton PA. Acoustic myography for
and muscle blood flow in fibromyalgia. European investigating human skeletal muscle fatigue. Journal
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71. 1991;71(4):1422-6.
10. Habacher G, Pittler MH, Ernst E. Effectiveness of 25. Merkens HW, Schamhardt HC, van Osch GJ,
acupuncture in veterinary medicine: systematic Hartman W. Ground reaction force patterns of Dutch
review. Journal of veterinary internal medicine / Warmbloods at the canter. American journal of
American College of Veterinary Internal Medicine. veterinary research. 1993;54(5):670-4.
2006;20(3):480-8.
11. Chvala S, Nowotny N, Kotzab E, Cain M, van den
Hoven R. Use of the meridian test for the detection
of equine herpesvirus type 1 infection in horses with
decreased performance. Journal of the American
Veterinary Medical Association. 2004;225(4):554-9.
12. Kim MS, Xie H. Use of electroacupuncture to treat
laryngeal hemiplegia in horses. The Veterinary

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Pearls from TVCM Practice
Feline Inappropriate Elimination and Lower Urinary

Tract Inflammation: Theoretical Background and Simple
Management with Acupuncture and Chinese Herbal
Medicine
Bruce Ferguson DVM, MS
ABSTRACT
Inappropriate elimination behavior, including urination and defecation, is relatively common in domestic cats kept as companion
animals. Such behavior is poorly tolerated by their human caretakers and is a common reason that owned, otherwise healthy
cats are given up for adoption, consigned to animal shelters, or euthanized. The traditional Chinese veterinary medicine
(TCVM) pattern for inappropriate urination (spraying/marking) is straight forward and is considered Heart Fire which is
related to chronic stress in the animal’s environment. Inappropriate defecation has a slightly more complex etiology which
leads to a TCVM pattern of Heart Yin Deficiency. There are two Chinese herbal medicines, Dao Chi San for Heart Fire and
Tian Wang Bu Xin Dan for Heart Yin Deficiency, which are non-toxic, readily available, inexpensive and rapidly effective
when coupled with appropriate husbandry changes.
Key words: Inappropriate elimination, inappropriate defecation, urine marking and spraying behavior, Chinese herbal
medicine, Dao Chi San, Tian Wang Bu Xin Dan, Heart Fire, Heart Yin Deficiency
fecal examination along with a good physical exam and

ABBREVIATIONS
if warranted, a more extensive medical work-up, to rule-
out a purely physiological cause for the aberrant behavior.
If those tests and exam are negative, the behavior is then
termed “psychogenic” in origin, or purely behavioral. In
In Australia and the USA, the most common reason this case, besides some basic behavior modification and
that owned, otherwise healthy cats are given up for possibly the use of feline pheromones, and/or anxiolytic
adoption, consigned to animal shelters, or euthanized drugs, the veterinarian is often unable to help. Moreover
is excessive aggression (e.g. attacking human family to further complicate the clinical picture, the average busy
members) or inappropriate elimination (e.g. on household veterinarian who performs a urinalysis, may mistakenly
furniture or owner’s clothing). 1 Inappropriate elimination believe that a urine sample with blood in it is equivalent to
may be either defecation or urination. Inappropriate a urinary tract infection. In fact, over 90% of the cases of
urination is commonly either misdiagnosed by presumptive feline lower urinary tract infection (FLUTD)
conventional veterinarians as a urinary tract infection, or which may present with hematuria, stranguria and dysuria
categorized as difficult to modify domestic cat behavior. are inflammatory, but otherwise sterile.1 It has been known
Inappropriate defecation is associated with unburied for almost two decades that cats tend to have a pathogenesis
piles of feces placed in areas of the owner’s environment of hematuria that is similar to human idiopathic interstitial
outside approved feline toileting facilities. cystitis which is exacerbated by chronic stress.1
Veterinarians are commonly called upon to help Inappropriate elimination has proven to be a difficult
owners of indoor domestic cats to deal with these condition to treat with commonly used western approaches
abnormal behaviors. The primary contribution of a such as antibiotics, behavior modification and anxiolytic
veterinarian is to at least perform a urinalysis and/or drugs. Recognition of TCVM patterns of imbalance in
these animals offers veterinarians a different and possibly
From: Bruce Ferguson, DVM, MS, St. Augustine Beach, Florida more effective approach to this disease condition. This
USA

paper summarizes basic feline behaviors and environments boxes that are unacceptable to the cat, learned aversion to
that lead to this condition, TCVM patterns associated a particular toilet box or dissatisfaction with a new type or
with inappropriate urination or defecation and successful brand of litter for the toilet box.6
treatment strategies using acupuncture and Chinese herbal A second category is human-related in which cats
medicine. can be exposed to chronic stress in the form of protracted
arguments in households or additions of new household
Marking Behavior in Cats members. The third category is animal-related such as
The domestic cat, Felis catus, evolved from the North too many cats in an indoor environment. In addition, in
African Wild Cat, Felis libyca. The North African Wild the author’s experience, one of the most common causes
Cat is a semi-solitary, highly territorial small carnivore1,2,3. is indoor cats which perceive outdoor cats roaming and
In order to demarcate territories and home ranges, this marking the property at night. These “rogue” cats are
wild ancestor of the domestic cat uses feces, normal commonly scent-marking which creates chronic stress for
urine, and specialized application of urine on vertical the indoor cats. The owners are unaware of the situation
objects, called “scent marking” or “urine marking”.4 because the problem occurs in the middle of the night,
Urine marking in this sense differs slightly from normal perhaps around a window or door, while the owners are
urination in that it is “sprayed” as a horizontal stream from fast asleep.7,8 The indoor cats, unable to travel outdoors to
the cat’s urethra onto vertical objects. This “spraying” scent-mark over the scents of the rogue intruders, live in a
behavior is found in both male and female domestic state of constant agitation.
cats, with “queens” (female cats) spraying at about a In the case of inappropriate urination, the pathogenesis
60% frequency and toms (male cats) closer to 90%.5 is fairly straightforward. Chronic stress disturbs the Shen
The behavior is partially hormonally-controlled, with and causes the Heart to develop Heat from stagnation of
about a 50% reduction in spraying behavior found after emotions (lack of joy). Since the Heart is the Emperor, it
castration of male domestic cats or ovariohysterectomy in is protected by the Small Intestine which drains the Heart
female cats. Cats also utilize selective deposits of feces to Heat to the foreleg Tai Yang Small Intestine and then to
demarcate territories and home ranges. This behavior also the hind leg Tai Yang Bladder which then eliminates the
appears to be similar to normal defecation, except that in Heat via normal urination. The problem arises when the
the case of normal defecation, the cat generally attempts to Heart Heat is chronic. Some residual Heat remains in the
bury the feces in some substrate (soil, cat litter, etc.). The Bladder which leads to a type of Lin Syndrome with Heat
defecation used for territorial marking is more likely to or Damp-Heat and hematuria. This is often exacerbated
be left in an unburied pile at a visually obvious location.5 by an affected cat consuming a dry kibble diet. This leads
These “marking” behaviors are both quite normal for the to a state of mild to moderate chronic dehydration which
wild ancestors of the domestic cat. Unfortunately, when creates the inability of the bladder to remove residual Heat
either of these behaviors occurs in a domestic cat kept due to inadequate fluid movement.
exclusively as an indoor companion animal to humans, When looking at the similar but different clinical
the behavior becomes completely inappropriate. Scent problem of inappropriate defecation, the TCVM pattern
marking by spraying on vertical surfaces (i.e. the walls or Bian Zheng is most commonly associated with Heart
of a home or the vertical edge of a couch) can both stain Yin Deficiency. Along with the stressors leading to
the materials as well as leave a tell-tale and unacceptable inappropriate urination delineated above, domestic cats
odor. Fecal scent marking by defecation in human shoes commonly are fed dry cat kibble which leads to chronic
and on clothing, for example, instead of in the “litter box” Yin Deficiency and dryness. The Heart Heat generated by
(a distinct container with a substrate which cats use for the companion cat having to tolerate other household cats,
defecation or urination) becomes intolerable as well. When unacceptable toileting facilities, possible human stressors,
either defecation or urination is outside of the acceptable etc., eventually consumes Heart Yin and coupled with
toilet area for a domestic cat, it is termed “inappropriate chronic dryness leads to Heart Yin Deficiency.
elimination/urination/defecation” or scent marking.
Treatment of Inappropriate Elimination with
Etiology and TCVM Pattern Traditional Chinese Veterinary Medicine
What can be said about the etiology and pathology As in conventional medicine, it is important to rule
of “inappropriate elimination” behaviors in traditional out urinary tract infection, renal failure with polyuria,
Chinese veterinary medicine (TCVM)? The etiology lumbosacral pain, and arthropathies which might inhibit
appears to be a chronic stressful event(s) related to the the cat’s ability to easily access the litter box. Once these
cat’s environment which fall into 3 primary categories. potential confounding or even causative issues are ruled
The first category is unacceptable toileting facilities which out, then the basic assumption left is that there is a Shen
can include a variety of situations such as too few litter Disturbance in the cat or cats who have inappropriate
boxes in a multi-cat household, poor sanitation of the elimination.6
litter box by humans leading to excessively soiled litter The simpler issue to address when treating

inappropriate elimination is “inappropriate urination” than ideal. First, it is difficult to be at the right place during
which has several components; stress reduction, the exact time of spraying so that the human can deliver
administration of Chinese herbal medicine, acupuncture a punishing or noxious stimulus when the cat sprays.
and behavior modification through training. If possible, Second, B.F. Skinner, the father of modern behaviorism,
stress should be eliminated or at least reduced in and “discovered” what we all already know to be true. The
outside of the home. This might include reducing the “punisher” becomes associated with the punishment, and
number of cats in the household, increasing the number thus we fear, avoid, or otherwise try to have minimal
of litter boxes in a multi-cat household, or finding a new contact with the punisher.10 A more ideal behavior modifier
partner who does not argue so much. It may also include is “remote punishment” which is delivered automatically
using a “live-trap” in the yard or garden to catch the by the environment at the exact time the behavior to be
“rogue” cat who may be disrupting the harmony of the punished occurs. In the case of spraying onto vertical
indoor cat’s world. surfaces, putting aluminum foil or clear “cling-wrap” on
The “sages of the ages” recognized the underlying the vertical object for just a few weeks usually does the
TCVM pattern for this condition and created a Chinese job. Whenever a cat backs up to the vertical surface and
herbal medicine to treat it.9 The primary offender cat begins to spray, some of the urine is splashed back onto
should be given Dao Chi Sana, which is an elegant the cat. Cats are generally very fastidious and dislike being
formula to reduce Heart Heat and calm the Shen (Table 1). sprayed with their own urine. This remote punishment
In addition, acupuncture or aqua-acupuncture with dilute technique usually stops the behavior, at least long enough
Vitamin B12 may be used at GV 20 or Da-feng-men to for the other interventions to take effect.
calm the Shen. Lastly, moist foods that increase the fluid What if inappropriate defecation rather than urination
moving through the urinary bladder, thus facilitating the is the problem? Again, the approach can be similar to
draining of the Heart Heat or Fire, should be substituted inappropriate urination such as stress reduction, Chinese
for dry and drying foods. herbal medicine and acupuncture. It is recommended that
If spraying on a vertical surface is the issue, in the cat owner experiment with new litter box substrates.
addition to the above changes, the client can add “remote Sometimes the substrates that are “earthen” materials
punishment”. Direct punishment, Type 1, done by a human (e.g. clay), although “natural”, tend to have too much
has at least two unfortunate problems that make it less fine powder associated with them and cats seem to dislike
Table 1: Ingredients and Actions of Dao Chi San

% English Name Chinese Pin-Yin Action
30% Rehmannia Sheng Di Huang Clears Heat, Cools Blood, Nourishes Yin
30% Akebia Mu Tong Promotes Urination, Drains Heart Heat via Small Intestine
20% Lophatheri Dan Zhu Yu Clears Heat, Promotes Urination
20% Glycyrrhiza Gan Cao Harmonizes and tonifies Qi
Table 2: Ingredients and Actions of Tian Wang Bu Xin Dan

% English Name Chinese Pin-Yin Action
30% Rehmannia Sheng Di Huang Clears Heat, Cools Blood, Nourishes Yin
4% Panax ginseng Ren Shen Tonifies Qi, Calms the Shen
8% Asparagus Tian Men Dong Clears Heat, Promotes Urination
4% Scrophularia Xuan Shen Enriches Yin, Clears Heat, Calms Shen
4% Salvia Dan Shen Tonifies Blood
4% Poria Fu Ling Calms Shen, Assists the Heart Qi
4% Polygala Yuan Zhi Calms Shen
8% Angelica sinensis Dang Gui Tonifies Blood
8% Schisandra Wu Wei Zhi Prevents leakage of HT Qi and Yin Fluids
8% Biota Bai Zi Calms Shen
8% Zizyphus Suan Zao Ren Prevents Leakage of Heart Qi and Yin
4% Platycodon Jie Geng Conducts Herbs to Upper Burner

inhaling that dangerous material. Recycled, compressed appropriate Chinese herbal medicine and addressing
newspaper (“Yesterday’s News”) is commonly accepted environmental stress; the TCVM practitioner can expect
by cats. The owner can also use the litter box to “capture” to treat this difficult clinical condition with high success.
the defecation behavior again, by placing it wherever there
is inappropriate elimination. Once the cat begins to use Footnotes
the litter box again, then move the litter box 0.5 meter/ a. Guide Out the Red Powder, Cathay Herbs, 5/5-15
day back to its original place in the household. Usually the Dunning Ave, Rosebery NSW 2018, Australia
cat’s elimination behavior will “follow” the litter box back b. Heavenly Emperor’s Tonify the Heart Pill, Cathay
to its old location. Herbs, 5/5-15 Dunning Ave, Rosebery NSW 2018,
If the above environmental modifications do not Australia
change the cat’s behavior, there is a need to ask if there is
something in the environment that is not apparent to the REFERENCES
human, yet is able to keep a cat from voluntarily using
the litter box. In other words, what accidentally trained 1. Neilson, J.C. (2014). Feline house-soiling:
aversions to litter boxes has been created? Is there a mild Elimination and Marking Behaviors. Topics in
electric current from an improperly grounded electrical Companion Animal Medicine. volume 19, 4:216-
appliance? Is there a periodic or seemingly random noise 224.
or some other sensory disturbance occurring, such as a 2. Sunquist, Melvin E.; Sunquist, Fiona (2002). Wild
washing machine close by? These potentially disturbing Cats of the World. University of Chicago Press. p. 10.
stimuli need to be found and either corrected, or simply 3. Germain, E.; Benhamou, S.; Poulle, M.-L. (2008).
move the litter box to a more peaceful setting. During such “Spatio-temporal Sharing between the European
a potentially disruptive change in the cat’s world, treatment Wildcat, the Domestic Cat and their Hybrids”.
should also include using aqua-acupuncture at GV 20. In Journal of Zoology 276 (2): 195–203
addition, for most “intense” cats that are Yin Deficient 4. Wade, Nicholas (29 June 2007). “Study Traces Cat’s
due to a dry kibble diet and poor water consumption, the Ancestry to Middle East”. New York Times (New
administration of Tian Wang Bu Xin Dana for 2-3 weeks York: NYTC). Retrieved 2 April 2008.
usually allows them to experience and tolerate the changes 5. Borchelt, P.L., and Voith, V.L. (1996). Elimination
that need to be made and helps restore balance (Table 2). Behavior Problems in Cats. Readings in Companion
Animal Behavior. pp. 179-190. Veterinary Learning
Conclusion Systems, Trenton, NJ.
In summary, two of the most problematic domestic 6. Baron, Alan; Stewart, C. N.; Warren, J. M. (1 January
cat behaviors that may disrupt positive social interactions 1957). “Patterns of Social Interaction in Cats (Felis
between companion cats and their caretakers are domestica)”. Behaviour 11 (1): 56–66.
inappropriate urination or urine marking and inappropriate 7. Case, Linda P. (2003). The Cat: Its Behavior,
defecation. When addressed from a TCVM viewpoint, Nutrition, and Health. Ames, IA: Iowa State
these are both types of Shen Disturbance. The subtype University Press.
of Shen Disturbance leading to inappropriate urination is 8. Kahn, Cynthia M.; Line, Scott (2007). Hollander,
most commonly Heart Fire and is rather easily addressed Joseph Lee, ed. The Merck/Merial Manual for Pet
by the Chinese herbal medicine, Dao Chi San, Guide Out Health. Merck.
the Red Powdera. The second subtype of Shen Disturbance 9. Chen, J.K., Chen,T. T., Beeb, S. and Salewski.
that is most commonly associated with inappropriate (2012). Chinese Herbal Formulas for Veterinarians.
defecation is Heart Yin Deficiency. If other husbandry pp. 272-274. Art of Medicine Press, Inc., City of
factors are adequately addressed, it is also easily treated Industry, CA.
with the Chinese herbal medicine, Tian Wang Bu Xin 10. Powers, R.B., and Osborne, J.G. (1976).
Danb, Heavenly Emperor’s Special Pill to Tonify the Fundamentals of Behavior. p. 91. West Publishing
Heart. Addressing the TCVM pattern, selection of the Co. USA.

Laser Acupuncture and Local Laser Therapy in

Veterinary Medicine with Overview of Applied Laser
Types and Clinical Uses
Uwe Petermann DVM
ABSTRACT
Local laser therapy or low level laser therapy (LLLT) has been enjoying increased popularity in recent years in both human
and veterinary medicine. It provides excellent healing in traumatized or infected tissue by increased energy (ATP) in diseased
tissues which have high energy requirements, provides peroxide clearance and restructures abnormal dense connective tissue
(e.g. in tendonitis). Acupuncture and LLLT are separate treatment modalities, but the synthesis of these two very effective
therapies allows local healing of diseased tissue with LLLT supported by acupuncture as an autonomous regulating therapy. It
is important to emphasize that the resulting “laser acupuncture” not only means the stimulation of the acupuncture point by the
laser beam instead of the needle, but that the local tissue healing effect of increased ATP production by laser light supplements
acupuncture as an integral part of the therapy. It is important to differentiate between the different types of lasers (continuous
wave versus pulsed wave) and the different features that are associated with laser effects such as: wavelength, power output and
resonance frequencies. When evaluating appropriate laser units for use in veterinary laser acupuncture, the pulsed lasers with
a 90 watt pulse peak and 904 nm wavelength are featured as they achieve high penetration depth with sufficient application
of laser photons in traumatized or infected tissue, without a thermal reaction in the tissue. For veterinary acupuncturists, the
addition of low level laser therapy and laser acupuncture can be used in addition to any thinkable acupuncture treatment which
gives additional options for achieving optimal results.
Key words: 904nm pulsed laser, impulse laser, low level laser therapy, acupuncture, laser acupuncture, horse, dog
by Michael Hamblin of the Harvard Medical School and

ABBREVIATIONS
the Massachusetts Institute of Technology (MIT), after
this laser effect was originally discovered in the 70s by
LLLT Low level impulse laser therapy
T. Karu and U. Warncke. 1,2,3,4
W Watts
Acupuncture and LLLT are separate treatment
TCVM Traditional
modalities, but the synthesis of these two very effective
Hz Hertz
therapies allows local healing of diseased tissue with
Fr Resonance Frequency
LLLT supported by acupuncture as an autonomous
Rf Reninger
regulating therapy. It is important to emphasize that
mW Milliwatts
the resulting “laser acupuncture” not only means
cw Continuous Wave
the stimulation of the acupuncture point by the laser
nm Nanometers
beam instead of the needle, but that the local tissue
Low level laser therapy (LLLT) has been used healing effect of increased ATP production by laser
approximately 25 years and is one of the most light supplements acupuncture as an integral part of the
underestimated therapies in veterinary and human therapy. Acupuncture and local laser therapy are two
medicine. One of the primary reasons for its slow therapies of equal value, each very effective by itself that
acceptance has been the use of a plurality of laser in combination lead to a total holistic therapy approach.
types which has led to widely differing study results.
The long-known fact that laser light can be converted Acupuncture and Laser Theory
directly into the cellular energy, ATP, was reconfirmed Acupuncture
Acupuncture has become an effective therapy for
From: Veterinary Acupuncture Clinic, Melle, Germany

many different disorders in both human and veterinary organs can be affected that result in organ Qi and Blood
medicine.5 It is an important component of traditional Stagnation and Deficiency (e.g. disease of the upper
Chinese veterinary medicine (TCVM) which is an entire thoracic vertebrae can affect the autonomic control of the
system of disease diagnosis and treatment based on 3000 heart).5
years of tradition, experience and research since the time of
Sun Yang (Bo Le), the father of veterinary acupuncture. The Laser
Channels and TCVM functional cycles play fundamental In Western medicine, the production of ATP (Qi) is
roles and are responsible for the smooth transport of Qi recognized as occurring primarily through mitochondria
(e.g. the sum total of the substances and physiological production (mammals) or photosynthesis (plants). In
activities of the body). The current understanding of plants, it is known that the transfer of light energy into cell
conventional anatomy and physiology can be useful to energy during photosynthesis occurs due to the chlorophyll
increase the understanding of ancient TCVM concepts. present in green leaves. Thanks to the chlorophyll, the
One form of Qi from a conventional perspective is plants can use sunlight directly for ATP genesis to produce
adenosine triphosphate (ATP) produced by the citric acid the necessary building blocks and the energy needs of cell
cycle (tricarboxylic acid and Krebs cycle) responsible for metabolism. Mammalian cells are not able to use sunlight
aerobic respiration within the mitochondria of the body. for energy generation and ATP generation is only possible
Aerobic respiration produces ATP that provides the body by oxidative phosphorylation in the respiratory chain of
with energy (Qi) for all life processes from the synthesis of mitochondria. Laser light is different from normal sunlight
enzymes to the relaxation of muscles. In all diseases, there in that it sends strong light clusters that due to laser
is a disturbance of the optimum flow of Qi that results in properties reach the redox system of the respiratory chain
a loss of balance between Yin (e.g. the parasympathetic in the mitochondria. According to Warncke, the absorption
nervous system) and Yang (e.g. the sympathetic nervous of laser light works via a so-called “antenna pigment”
system).5 which links it to the respiratory chain in the mitochondria.
When a disease irritates afferent nerves, pain and 3,4
The system takes up light in the average wavelength
reflex contraction of local muscles and blood vessels range of 900nm with a range of 600 to 1400nm.
restricts the free flow of Qi (ATP) with resulting Blood According to Hamblin, Karu and Warnke, the
and Qi Stagnation. The Qi/Blood Stagnation results in conversion of laser light into cellular energy (ATP),
Deficiency of Qi (ATP) and Blood (nutrients) to local particularly benefits diseased cells and cells in
tissues. Since Qi (ATP) is necessary for muscle relaxation, pathologically altered tissue, which have a particularly high
a vicious cycle of contraction and vascular occlusion energy demand1,2,3,4. Likewise, a strong boost of energy by
occurs and further worsens the Stagnation of Qi and the laser light in nerve cells at an acupuncture point can
Blood. Further, autonomic reflex connections to internal generate an action potential similar to acupuncture needle
Figure 1: Laserpen (70 mW, 785 nm, cw Figure 2: 8 Hz, oscillating between 50mW and 100mW; 4 Hz oscillating
laser), Reimers & Janssen, Berlin, Germany. between 0 and 50mW; mW = milliwatt

stimulus effect1,2 . This fascinating discovery that the laser continuous wave (cw) or impulse (pulsed) wave. The
light is directly converted to ATP in the flavoprotein- continuous wave (cw) laser (Figure 1), has a continuous
metal- redox- system of mitochondria, was confirmed output of power. As an example, class 3B lasers (cw)
and continued by investigations of Michael R. Hamblin1. would have a continuous wave output of 30-500 mW
Michael R. Hamblin is Associate Professor at Harvard (milliwatt) (Figure 2) and class 4 (cw) would have a
Medical School and director of the Wellman Center for continuous wave output of 1-8W (watt). In the second
Photomedicine at Harvard Medical School and MIT. This type of laser, the impulse laser, also named pulsed laser or
confirmation from a well-known scientist at a prestigious superpulsed laser (Figure 3), the laser energy is emitted in
institute has led to the increasing use and popularity of the form of single very strong light pulses. The individual
laser technology, especially in the USA. light pulses have power spikes from 30-100W, about 1000
This direct provision of energy in the form of times more power output than a continuous wave laser in
additional ATP provides an excellent local effect with the same class. The emitted high energy pulses, however,
laser therapy. The laser light provides cells with urgently are of very short duration, only 200 nano-seconds each
needed ATP (similar to increasing local Qi in acupuncture), (Figure 4). These quick high pulses of energy provide
which provides clinical effects such as relaxation of much greater tissue penetration by the laser light, but due
muscle spasms associated with abnormal vertebral body to the short duration of the light flashes, even at high pulse
alignment (spondylosis), the maintenance of depleting frequencies of modern lasers (up to 40,000Hz), no thermal
organ functions and repair of wounds. The additional or even coagulating effect in tissue has been seen11,12. This
energy aides in removal of wound debris which blocks has the great advantage of deep tissue penetration without
synthesis of new components for healing and assists in emitting so much energy that it heats up or burns the tissue
restructuring damaged tendon tissue with the incorporation at the surface as may occur in continuous wave lasers.
of new collagen fibers6. Other important properties of
infrared laser light is its ability to increase clearance of Laser Classes, Laser Power and Wavelength
peroxide radicals which provide an outstanding effect In addition to consideration of the laser type
against infectious organisms both in wounds and organ (continuous vs pulsed), there are 3 additional defining
tissue.7,8 If the therapist has sufficient experience, laser features to consider when comparing lasers. These are
therapy can be used without any adjuvant therapy, such power output (mW or W), wavelength (nm) and resonance
as antibiotics. This independence from antibiotics is of (Hz). Lasers are divided into a number of classes (1, 2, 3A,
particular importance when it comes to infections with 3B, 4) depending upon the power output (energy) of the
antibiotic resistant pathogens. A final characteristic of the beam and the wavelength of the emitted radiation (Table
laser light that supplements the increased energy to tissues 1). Laser power (mW/W), is mainly responsible for the
is the promotion of blood circulation, specifically in penetration depth and the energy output per time (Joule),
microcirculatory disorders, such as in equine laminitis.9,10 which both together are responsible for a sufficient number
of laser photons being transported deep into the tissue
Laser Selection where the energy is needed. Wavelengths suitable for
Laser Types veterinary laser therapy operate primarily in the infrared
There are 2 types of lasers based on wave production: range (780-1400 nm) which is an important criterion for
Figure 3: Physiolaser (2x 90 Watt, 904nm, impulse Figure 4: Impulse laser with 100Watt pulse peak power (the
lasers), Reimers & Janssen, Berlin. Germany. width actually is smaller than shown in the figure).
depth of tissue penetration (along with power output) and by the Fr of the light pulses. The three most important
is necessary for stimulation of acupuncture points. 1,2,3,4 currently used series of resonance frequencies are Nogier,
Bahr and Reininger (Table 2).13,14,15
Resonance The most commonly used laser Fr series are those
In biology, resonance frequencies are of extreme according to Nogier13,. The French acupuncturist Dr.
importance and largely determine the therapeutic effect of Paul Nogier, who discovered the laser for the use of
laser treatment. The continuous laser beam is modulated acupuncture, also developed these frequencies (Table 2).
by a specific resonance frequency (Hz) while in the He established a range of 7 frequencies named A ‘, B’, C
impulse laser, the resonance frequency (Fr) is achieved ‘…G ‘. The Fr A’ (292Hz) is the most important one, since
Table 1: Laser classes are defined by wavelength and power output
Laser Classes Power Output* Wavelength Example

Class 1 Laser Printer
Class 2 0-1 mW Visible light Laser Pointer
Class 3 A < 5 mW Visible light 400-700 nm Laser sights-firearms
Class 3B 5-500 mW Infared LLLT, Laser Medical
700-1400 nm Device
Class 4 >500 mW Infared 700-1400 nm Laser Medical Device
*Continuous Wave (cw); pulsed wave peaks have higher power output
Table 2: Comparison indications and attributes of the Nogier, Bahr and Reininger frequencies used for low-level impulse laser
therapy and laser acupuncture
Nogier Frequencies
Frequency A’/A’’ B’ C’ D’ E’ F’ G’
Hz 292/37376 584 1,168 2,336 4,672 9,344 18,688
Tendonitis,
Wounds, Tendonitis, Mandibular
arthritis, Cerebral
inflammation, arthritis, Nerve and joint and
fractures, Acupoints of cortex and
Indications irritable foci fractures, spinal cord subcortical
all body the feet mental
in body and organ diseases brain
acupoints disorders
teeth acupoints disorders
except feet
Bahr Frequencies
Frequency 1 2 3 4 5 6 7
Hz 599.5 1,199 2,398 4,796 9,592 19,184 38,368
Central
Deep tissue layer, Opening
tissue layer, Surface Conception
“deep” points acupoints
Indications “subsequent” tissue Omega Vessel,
(source of illness, of the Governing
and points structures, Du “hidden”
related to the Eight Ex- Vessel
Attributes (related to the Omega Ren Channel foci in
sympathetic traordinary
parasympathetic Channel teeth
system) Channels
system)
Reininger Frequencies (Meridian Frequencies)
Channel LIV ST HT PC LI GB KID BL SP TH SI LU
Hz 442 471 497 530 553 583 611 667 702 732 791 834
*Used for treatment of acupoints on specific Channels and topical treatment of related organs; LIV=Liver, ST=Stomach,
HT=Heart, PC=Pericardium, LI=Large Intestine, GB=Gallbladder, KID=Kidney, BL=Bladder, SP=Spleen, TH=Triple Heater,
SI=Small Intestine, LU=Lung Channels

it is the resonance used to treat inflammation (e.g. wound is applied (Laserpenb). The impulse lasers (90W) can
infections, joint inflammation, organ inflammation). provide sufficient energy to stimulate acupuncture
Modern high tech lasers provide the high Fr A’’ with points, support tissue regeneration, wound healing and
37,376 Hz which provides 128 times more energy per infection control even at considerable tissue depth, such
time than Fr A’. Frequency C is the “orthopedic” Fr and as needed for equine work without generating excessive
is used both for treating orthopedic acupuncture points damaging heat as can be seen with Class 4 units.8,10,11,12
in auricular medicine (ear acupuncture) as well as the
local treatment of joints and the entire spinal column Treatment Principles
(especially blockages of the spine). Frequency E is the Laser energy and energy dose
resonance used to treat nervous tissue and spinal cord. When treating a diseased tissue locally (LLLT) or
The Bahr frequencies, also a series of 7 frequencies B1, performing laser acupuncture; careful consideration should
B2, B3, …B7 are very important for the pulse controlled be given to the amount of energy that needs to be supplied
acupuncturists for acupuncture diagnosis and therapy.14 to a tissue for optimal tissue effect (energy dose). Laser
Frequency B5 is also important for the local treatment of energy (Joule), is the amount of energy that is delivered
inflammation as an alternative to Fr A ‘ in impulse lasers to the tissue or to the acupuncture point. The absorbed
that do not have the high Fr A’’. Frequency 7 (299.5 Hz) is energy in the target area, which provides usable energy for
a resonance Fr for treatment of another kind of disturbing the organism, is determined by 3 factors: the laser power
foci in the teeth, so called “hidden disturbing foci”. Here in watts, the depth of the pathological process and the
again, similar to A’ and A’’, only new high tech lasers will irradiation time. Based on the author’s experience, it can
have high Fr 7’ with 38,386 Hz, which is 128 times more be assumed that an optimal effect on superficial structures,
energy per time compared to Fr 7. such as an acupuncture point with an impulse laser (90W
The Reininger frequencies have specific resonance peak pulse power), is achieved with an exposure time of
to the individual Meridians and organs which they are approximately 20-40 sec. This corresponds to an applied
representing.15 Thus, the Reininger Fr, Heart, is used energy of 0.3 to 1 Joule. In deep wounds, tendonitis,
for optimal treatment both for all acupoints of the Heart spondylosis, kissing spines, arthritis, laminitis, organ
Meridian as well as for the local treatment of the heart diseases (topical transcutaneous treatment) or deeper
muscle. The same applies to all other Meridian frequencies, infections, there will need to be an increase in the duration
such as the Kidney, Liver, Lung etc. of therapy to 2-3 minutes per location.
It must be emphasized that with an impulse laser, the
Selection of Laser Unit for Veterinary Clinical Use emitted energy dose depends on the number of emitted light
When choosing a therapy laser, it is very important pulses, since each pulse represents a unit of energy. There is
to identify an individual veterinarian’s needs and to a significant difference between the “normal” infection Fr
select a laser best suited for the clinical cases it will be A’ with 292 Hz and the “high” infection Fr A’’ with 37,376
used for. Suitable lasers for LLLT primarily belong to Hz. The energy output per time in a 90W impulse laser, at
Class 3B lasers and more recently, Class 4 lasers, which the higher Fr A’’ is 128 times higher than the “normal” Fr
differ in average energy output. The Class 3B lasers have A’. To apply 10 Joule of energy with Fr A’ (292 Hz) will
a maximum of 500milliwatts (continuous wave power, require 32 min. The same amount of energy provided by
or averaged power for impulse lasers). Class 4 lasers the laser with Fr A’’ (36,368 Hz), will require only 26 sec.
(continuous wave) are characterized by high power (up to It is particularly important to point out that the therapy
8W) and were designed initially for laser surgery. Since success described by the author has been achieved
there is “light off” between the single pulses of high solely with a 90W, 904nm pulsed lasera with the “high”
intensity in pulsed lasers, over time the average emitted inflammatory Fr A’’ (38,386 Hz), because only this
energy from the 3B laser is lower than in Class 4 lasers. laser unit transports sufficient energy into the depth
Although Class 4 lasers have a higher continuous wave of the tissue in an acceptable treatment time without
power output when compared to Class 3B impulse lasers damaging overlying tissue layers. The applied laser
(8W versus 500mW), the impulse laser can produce the energy is dependent on the pulse Fr between 1-10 joules
same amount or greater power during their peak spikes per localization11,12. The difference between the various
(90W versus 8W). species is of less importance, however, a wound can be
Other considerations for suitable LLLT lasers is deeper in a horse than a dog and the treatment of an organ
to look for wavelengths in the infrared range (780- needs more time in large animals than in small animals.
1400) and units that can program Nogier and Bahr A continuous wave laser is used solely by the author
resonance frequencies (usually not available in Class for the local treatment of eye diseases such as equine
4 lasers).1,2,3,4 In the author’s experience (28 years of recurrent uveitis, (ERU), corneal injuries, corneal ulcers
daily use), the 904nm (wavelength), Class 3B impulse etc. The applied energy in these conditions is about 1
laser (super pulsed to 90W), Physiolasera, gives the best joule, which is an equivalent for a 100mW continuous
benefits. The only exception is the local treatment of wave laser therapyb for about 20 sec. In contrast to the
the eye, where the continuous wave laser with 70mW body tissues, the eye is optically transmissive and the
cellular need for laser energy is at the surface of this sec. The continuous wave laser emits 1 Joule (100mW), the
transparent medium. Here, high wattage is not needed as pulse laser, depending on the therapy Fr about 1-10 joules.
it may penetrate too deep. The pulse laser with high pulse
powers would virtually “shoot into the void” of the eye. Laser acupuncture techniques
Both continuous wave and pulsed laser units are Laser acupuncture differs from pure LLLT in that
equally effective for laser therapy of acupuncture points additionally to the local effects of laser use, the clinician
as long as various resonance frequencies are available for is providing the disease treating benefits of acupuncture.
selection. For optimum treatment of an acupuncture point, A few very effective acupuncture points, which are
regardless of the type of laser (100mW continuous wave easy to use are presented (Table 3). These are acupoints
or impulse laser 90W), the treatment time is about 20 to 30 used regularly for some common conditions5. Every
Table 3: Acupoints, their indications, treatment time and strength, wavelength and frequencies of the impulse laser used in
the case studies
TCVM PCLAC Laser Strength Laser wavelength Pulse Frequency Treatment time
Acupoints
indication indication (Watt) (nm) (Herz) (seconds)
Tonifying
KID-7 Kidney point 90 904 Rf. KID (611) 20
point KID
Master pain
LI-4 Thalamus point 90 904 Rf. LI (553) 20
relief
Tonifying
LU-9 Blood perfusion 90 904 Rf. LU (824) 20
point LU
LIV-13
LIV Pituitary gland 90 904 Rf. LIV (442) 20
Tonifying
LIV-8 Liver point 90 904 Rf. LIV (442) 20
point LIV
Luo- TH and
TH-5 Thymus point 90 904 Rf. TH (732) 20
opening point
Back-Shu point Back-Shu point,
Nogier freq. C
BL-23 KID,Yang Sympathetic 90 904 20-40
(1168)
aspect KID aspect KID
Back-Shu point Back-Shu point,
Nogier freq. C
BL-18 LIV,Yang Sympathetic 90 904 20-40
(1168)
aspect LIV aspect LIV
Nogier freq. A’’
BL-1 Local Eye Local Eye 90 904 20
(37376)
ST-1 Local Eye Local Eye 90 904 20
(37376)
TH-23 Local Eye Local Eye 90 904 20
(37376)
Auricular point Nogier freq. A’’
GB-1 Local Eye 90 904 20
of the eye (37376)
SP-2 Tonifying Anabolic master
90 904 Rf. SP (702) 20
point SP point
SP-4 Luo- SP and
Interferon 90 904 Rf. SP (702) 20
opening point
GB-41 Opening point Prostaglandin 90 904 Rf. GB (583) 20
GB E1 point
SI-3 Opening point Master point 90 904 Rf. SI (791) 20
SI spasm
nm = nanometers, hertz = cycles per second; Nf. = Nogier frequency, Rf. = Reininger (meridian) frequency,
PCLAC = Pulse Controlled Laser Acupuncture Concept

experienced acupuncturist can combine LLLT with any of infected wounds or disturbing foci, throughout the
acupuncture treatment concept that one has in mind, even experience period of the author, no adverse effects or
if the acupoints are treated by a needle or by the laser with burns have occurred.
the appropriate resonance Fr. Class 4 lasers should have very strict precautions,
The laser should always be placed directly on the skin especially regarding the protection of the eyes. The risk
of the irradiated area, so that as little energy as possible of skin and tissue burns are higher with these lasers, since
is lost. The acupuncture points should be treated on both much of the laser energy is absorbed in the outer skin.
sides of the body. Local Laser Therapy, or the topical The author is aware of a number of cases of significant
(local) irradiation of the diseased tissue varies depending burns to the backs of horses with these units. Such events
on the size or volume. According to the depth of the contribute neither to treatment benefit nor to a better
location of the disease and according to the type of tissue acceptance of laser therapy.
and the nature of the pathological process, Fr and applied
energy have to be adapted to the body’s needs. The same Laser Use in Clinical Cases
is true in the case of local treatment of organs (Table 4). Wound treatment (Acute or Surgical Wound)
During the treatment of open wounds, the laser
Handling Precautions for Class 3B and Class 4 Lasers should always be wrapped (plastic), to both protect
Although, in the author’s veterinary clinic, there the wound against contamination from environmental
have been no accidents and there appears to be very bacteria and to protect the laser from contamination by
little accident potential to the eyes with Class 3B pulsed wound bacteria. If possible, during wound treatment, the
lasers, there are certain prescribed safety regulations for laser should be placed directly on or inside the wound
handling lasers, which naturally should be followed. In all (even to a depth of 1-2 cm). To promote wound healing in
lasers recommended by the author: Laserpenb (70/100mW non-infected wounds, treating the wound edges for 10 sec/
cw laser) to treat the eye and Physiolasera (90W pulsed cm with Nogier Fr B is sufficient. To prevent infection, the
lasers) for tissue therapy, the emitted laser beams have a wound depth can be treated for an additional 10 sec with
divergence of about 10 °. This means that no focused laser Fr A’’. In addition to local laser therapy, the acupoint, SP-2
beam can hit the eye, where it can cause injury. Still, wearing (anabolic master point wound healing/tissue building) is
laser safety goggles is mandatory and people present in the treated for approximately 20 seconds (100mW cw or 90W
room should wear such glasses. The treatment room should pulsed laser). This significantly improves wound healing
be free of mirrors and reflective metal surfaces to prevent and avoids secondary infection of surgical wounds which
any conceivable damage to the eyes of owners and staff. provides fast and stable wound closure and dehiscence
In the topical treatment of tissue or laser acupuncture can be reliably avoided. Tissue damage from laser therapy
using Class 3B cw laser or Class 3B impulse laser with with these lasers is unknown2,6
unfocused laser beam, there is no risk of skin burns. In
patients with strongly pigmented skin, particularly after Wound treatment (Infected Wounds)
prolonged laser exposure at high frequencies (Nogier E Laser acupuncture, in the author’s experience, is
‘, F’, G ‘, A’’, Bahr 5, 6 and 7’), if the animal is showing the treatment of choice for infected wounds; no matter
some discomfort; the treatment of that particular point whether it is an acute infection, a contaminated wound
is interrupted for a few seconds, until the animal is associated with trauma or a 6 month old castration wound
comfortable and then treatment with the desired energy with antibiotic resistance and revision surgery failure.8
is resumed. Here too, even at very intensive therapy Laser acupuncture and LLLT is extremely reliable, even in
Table 4: Laser Treatment Times and Energy Dose*

Location Energy Dose Treatment Time
Acupuncture Point 0.5-1 Joule 20 sec
Superficial Wound 2-3 Joule/cm2 20-40 sec
Deeper Wound Infected Wound 6-10 Joule/cm2 30-60 sec
Joint 4-16 Joule/location 30-120 sec
Tendon 4-16 Joule/location 30-120 sec
Spinal Cord 4-16 Joule/location 30-120 sec
Disturbing Focus-Scars 4-8 Joule/location 30-60 sec
Disturbing Focus-Teeth 8-20 Joule/location 60-120 sec
*Using Class 3B, 90W, Pulsed Laser (Physiolaser)
hopeless cases. The treatment consists of local treatment Rf Meridian frequencies. This treatment was continued
of the wound with Fr A’’ with an applied amount of energy by the owner with an identical laser at home every day
of 10-50 joules, depending on the size and the depth of the using the same protocol. After 1 week, the swelling was
wound. This corresponds to a treatment time of about 1-5 significantly reduced with the wound demonstrating
minutes with a 90W impulse laser. The major acupuncture less pain and reddening. The erysipelous discoloration
points to use are SP-2 (anabolic master point) to promote had almost disappeared. After another week of the same
wound healing, SP-4 (interferon point) and TH-5 (thymus therapy, the infection was healed. At a clinical recheck 14
point) for 20-30 sec with the corresponding Reninger (Rf) days later, the surgical scar was barely visible (Figure 6).
Meridian frequencies to fight infection. Depending on the
severity of the infection, the wound must be treated daily Muscle and tendon injuries
to 2 times a week until complete healing. The treatment Laser acupuncture and LLLT is particularly effective
intervals can be adapted to the healing process. in the treatment of muscle and tendon injuries. The
problem with acute and chronic tendonitis of the horse,
Case example: MRSA (Methicillin Resistant and ligament and muscle injuries in dogs and horses is
Staphylococcus Aureus) infection after castration of a that damaged tendons and muscle fibers are repaired by
2-year-old Bernese Mountain dog connective tissue. This dense, firm connective tissue is
A 2-year-old Bernese Mountain dog was presented inelastic and therefore extremely vulnerable to re-injury.
with a 15-20 cm purulent swelling of a castration wound. The elastic and muscle fibers cannot penetrate the scarred
The dog had been neutered 4 months previously. Within a area to a sufficient extent to restore the original structure
few days following surgery, a fistulous wound was noted. and function of the tendon or muscle.6 Low Level Laser
Methicillin Resistant Staphylococcus Aureus was cultured Therapy (LLLT) plays a fundamental role because it
from the tract. Antibiotic therapy with various antibiotic represents, in the author’s experience the best way to
cocktails, anti-inflammatories, wound drains along with loosen and restructure the scarred dense connective tissue,
additional surgical procedures for 4 months proved so that normal collagen fibers can be incorporated.15
ineffective in controlling the infection. The markedly
swollen, fluctuating wound was very painful, severely Case example: Complete rupture of the superficial
reddened and was characterized by an erysipelatous-like flexor tendon in a Trakehner mare
skin discoloration (Figure 5). The treatment was simple. An 18 year old Trakehner mare was presented with
In addition to 3 minutes local laser therapya with Fr a complete rupture of the superficial digital flexor tendon.
A ‘’ (37,376 Hz); acupoints for immune stimulation were Examination of the open wound revealed a tendon rupture
treated for 30 sec: TH-5 and SP-4 with the corresponding with the two ends of the tendon at a distance of about 6 cm.
Figure 6: The 2-year Bernese Mountain dog at clinical

recheck 14 days after laser acupuncture and LLLT. The dog
Figure 5: A 2-year-old Bernese Mountain dog presented with is culture negative for MRSA and the surgical scar is barely
a 15-20 cm purulent swelling of a castration wound. The dog visible.
had been neutered 4 months previously. Within a few days
following surgery, a fistulous wound was noted. Methicillin
Resistant Staphylococcus Aureus was cultured from the
tract. The markedly swollen, fluctuating wound was painful,
severely reddened and was characterized by an erysipelatous-
like skin discoloration. This photograph taken after the 2nd
revision surgery before laser treatment.

The wound was too contaminated for surgical treatment, were also treated daily with Fr Bahr 5. The wound was
therefore, laser acupuncture was preferred because of the cleaned daily with a lavage fluid (2g Rivanol® +50ml
excellent antibacterial, wound and tendon healing effects. Echinacea tincture, 50 ml Arnica tincture and 50 ml
Daily treatment consisted of intense irradiation of the Calendula tincture per liter) and kept under bandage.
entire wound surface, particularly the tendon ends with Except for a tetanus booster, no further therapy was
the anti-inflammatory laser Fr Nogier A’’ for 5 minutes carried out with antibiotics or anti-inflammatory drugs.
with a 5 x 30W pulsed multicluster probea (Figure 7). Despite intensive irradiation with anti-inflammatory Fr A’’,
Acupuncture points to promote wound healing (SP-2, initial swelling of the surrounding tissue and granulation
with Reininger Fr for the Spleen) and TH-5 and SP-4 tissue formation could not completely be suppressed. Two
(to remove necrotic tissue and control inflammation) weeks after starting laser acupuncture, a large wound and
Figures 7 and 8: An 18 year old Trakehner mare presented with complete rupture of the superficial digital flexor tendon.
Examination of the open wound revealed a tendon rupture with a 6 cm distance between the two ends of the tendon. The
wound was too contaminated for surgical treatment, therefore, laser cupuncture was preferred because of the excellent
antibacterial, wound and tendon healing effects. Tendon wound at the beginning of therapy (left photo) and at the end of laser
acupuncture and LLLT (right photo).
Figure 9: Modern impulse laser penetrating the hoof wall in a horse with laminitis. The cast has drilled holes of 8mm each
for placing the laser tip directly on the hoof wall.
swelling (8 cm diameter) was still present. is usually a visible improvement in gait within hours.
The horse had very little pain during therapy so Local treatment is applied with alternating Nogier Fr
treatment was continued at the author’s clinic in 2 day A’’ and Fr B. This option can automatically be selected
intervals with slow but steady progress noted. After a in the Physiolasera or the change can be made manually
total of 8 weeks, the external wound was closed and the during treatment in 10-second intervals between the two
mare was dismissed. Further treatment was continued by frequencies. The central part of the front hoof wall is
the owner with the same treatment protocol for another mainly treated (area of 3x5 cm), Figure 9. One can also
6 weeks until healing was completed. The healed tendon use a 5 x 60w multi-cluster probea with 5 laser diodes,
continued to improve to the point that almost normal which considerably shortens the individual duration of
physiological strength was achieved (Figure 8). The therapy. If there is a risk of sloughing hoof, the affected
mare was slowly started under saddle at a walk and as part of the coronary band should be intensively treated
improvement and comfort continued, she slowly returned locally with Fr A’’ and B’.
to former activities. Acupuncture points to use in these cases are LIV-
8 (Rf Meridian Fr Liver), SP-2 (Rf Meridian Fr Spleen/
Equine laminitis pancreas), KID-7 (Rf Meridian Fr Kidney) and GB-41 (Rf
Laser acupuncture can be an effective treatment Meridian Fr Gall Bladder) for 20 sec each on both sides. In
of laminitis in all stages and degrees of rotation10. In the case of metabolic syndrome or Cushing Disease, treat
these cases, the synergism of the acupuncture effect and the pituitary locally between the TMJ (temporal mandibular
local laser therapy is particularly clear. The laser locally Joint) and ear with Rf Liver Fr and A’’ for 1 minute
improves the microcirculation and peroxide clearance each on both sides and the acupoint LIV-13 for 20 sec.
which reduces pain and inflammation quickly 9,7 There
Figure 10 and 11: An 11- year-old Trakehner mare developed severe acute laminitis at the beginning of the grazing season.
Hoof conformation before (left photo) and after laser acupuncture (right photo).
Figure 12 and 13: Same 11 year old Trakehner with significant rotation of the left front coffin bone before start of laser ther-
apy (confirmed by radiographs, photo on left). Photo on right of radiograph of the left hoof after laser acupuncture.

Case Example: Severe acute laminitis with coffin bone ATP input) which the energy insufficient muscle urgently
rotation in an 11-year-old Trakehner mare needs to relax and, second, at the same time, the laser
An 11- year-old Trakehner mare developed severe light improves microcirculation in the contracted blood
acute laminitis at the beginning of the grazing season. vessels.1,2,3,4,8 This is accomplished by application of the
Despite intensive therapy for 1 week in a veterinary laser on both sides of the vertebral blockage with Fr Nogier
hospital, there was significant rotation of the left front C, both dog and horse, about 1 minute per location. The
coffin bone (confirmed by radiographs) (Figure 10). The third effect of the laser is acupuncture stimulus of the back-
mare was severely lame on both front feet with non-weight Shu point which acts directly on the “Segment Regulatory
bearing lameness on the left front (Figure 12). Treatment complex“ and immediately interrupts the vicious circle
protocol included 4 treatments at intervals of 3 to 4 days and reduces muscle tone.16 The fourth influence of the
with laser acupuncture and LLLT at the author’s clinic. laser is calming the irritable state of the nerve root. The
Local treatment of the lamina to improve circulation optimal therapeutic Fr in this case is Fr Nogier E with
and reduce inflammation was performed with a multi- affinity to nerve tissue, which is applied for 30 sec. Thus,
cluster probea. Additionally, acupuncture points to support stimulation by the laser performs 4 synergistic treatments
Liver and Kidney function (LIV-8, KID-7) and GB-41 at the same time.
(prostaglandin point) for pain and inflammation were Acupuncture points for support are LI-4 (master
used. After 14 days, the mare’s lameness was significantly pain relief point) 20 sec with Fr LI (Reininger) and GB-
reduced and she was able to trot. The horse was released to 41 (master point rheumatic pain) 20 sec with Fr Bahr
go home and treated by the owner with the same treatment 5. With this simple therapy more than 1000 horses and
protocol for the next 2 weeks. Expert farrier care of the dogs have been successfully treated by the author.17,18
hooves addressed abnormal growth by trimming the left In addition, an experienced acupuncturist can add any
toe as much as possible frequently. Four weeks after laser additional complimentary acupuncture points, depending
acupuncture was started, the mare was sound and within on the needs of the patient and TCVM pattern diagnosis.
6 more weeks, she was put into full training. After a total
of 4 months, radiographs were taken again and showed Acute and chronic lameness
that the rotation of the coffin bone was almost completely If the exact inciting area of a lameness is known, a
resolved and the newly formed hoof wall oriented almost simple treatment of local laser therapy and acupuncture
parallel to the coffin bone again (Figures 11 and 13). The is possible. This involves both acute lameness as sprains
mare is performing successfully in dressage now and at or contusions and chronic lameness, where a combination
1 year recheck, without any further treatment, is sound. of muscle and tendon disorders, insertion-desmopathy
and joint irritation have led to osteoarthritis. In all
Back pain, spinal disorders and spondylosis lameness, consideration should be from the perspective of
The almost ubiquitous back pain in horses and acupuncture as well as from the perspective of physiology.
spondylosis in dogs are predominately associated with The close connection between the above mentioned nerve
small displacements of the facet joints of focal sections root irritations and the segment regulatory complex may
of the spine (vertebra blockages). These blockages can form a pathologic tonus of muscles and tendons which
evoke nerve root irritation in the affected segment which secondarily harms the peripheral joints. This means that
stimulates the “segment regulatory complex” inside the assessment should also consider underlying back and even
posterior horn of the spinal cord.16 This creates permanent neck issues when treating a lameness. A similar treatment
muscle contraction, which locks the vertebral bodies in a as mentioned above can also be used for treatment. The
position of misalignment. It also leads to contraction of local joint pain and inflammation is treated with Nogier
blood vessels and thereby reduced blood flow resulting in Fr C’ and Nogier A’’ in combination. Modern lasers offer
acidosis. Here arises the disastrous vicious circle of pain, the setting of alternating frequencies, or one can treat with
duration of contraction, malnutrition and acidosis, from each of the two frequencies sequentially, each 30 sec with
which the patient cannot recover. The outcome in acute Fr C and 15-20 sec with Fr A’’. Supporting acupuncture
cases can be “tying up” in horses and in chronic cases if points are the same as in back problems, GB-41 and LI-
the spasm and the pain becomes permanent, the syndrome 4. Any acupuncture treatment can be applied additionally
of “kissing spines” occurs. In dogs, a similar genesis to this simple concept depending on a clinician’s
results in disc disease and spondylosis. own experience as to what has worked well before.
A very simple, but nevertheless very effective
treatment is the use of laser acupuncture.17,18 In the horse, Eye diseases
simple palpation of the back for pressure sensitive areas Treatment of eye diseases is another area where there
identifies painful areas. The horse will clearly show where is clear benefit associated with the use of laser acupuncture.
the pain is localized.19 With the use of the laser, a clinician Regularly treated eye diseases are corneal injuries and
now has 4 excellent treatment effects at once. First, local corneal ulcers, as well as immunologic eye diseases, such
LLLT, very effectively provides direct local energy (Qi / as equine recurrent uveitis. For local treatment of the eye,

a 70mW cw laserb (Figure 2) is used instead of the impulse TH-23. Acupoints LIV-8, TH-5, GB-41 were treated with
laser. This is in contrast to all other local laser treatment. the Rf Meridian frequencies. In addition, the ulcer was
In corneal ulcer or corneal injury, the beam path is applied treated locally with direct but slightly tangential continuous
strictly tangentially to the injured areas of the cornea. In wave laserb (Fr A) for 30 sec. The medical pretreatment
acute inflammation, Nogier Fr A is used for 30 seconds; was continued unchanged in this case. At intervals of 4-5
for regeneration, such as corneal injuries, Nogier Fr B is days, the eye was treated by the author with the treatment
used. protocol just described, and between sessions in the same
If the inflammatory process also includes the interior manner by the owners. At the first clinical recheck after 4
of the eye, as in equine recurrent uveitis, the laser beam days, there was an incipient organization of the ulcer with
can also be applied for 20-30 seconds up to an angle of marked pannus and absorption of the reddish secretion in
approximately 45° to the inner eye. It is important to avoid the anterior chamber. The pain was significantly decreased
irradiation of the retina directly. No adverse side-effect and the pony more readily tolerated the application
from treatment of the eye has been observed, however it is of ophthalmic ointment. After another 4 days, further
prudent to be cautious. improvement was obvious and antibiotic treatment was
There are 4 local acupoints around the eye, BL-1, ST- discontinued. After 18 days of treatment, following the
1, TH-23 and GB-1. GB-1 is identical to the eye point of first acupuncture treatment, the eye showed complete
ear acupuncture. Other points are LIV-8, GB-41 and TH-5. regeneration with no residual dysfunction (Figure 15). The
All are treated for about 20-30 sec with the corresponding original perforation was now a white, non-reactive scar,
Rf meridian Fr. For this purpose, both the pulse lasera and which did not create any visual limitations for the pony.
the continuous wave laserb may be used. The treatment
time for both lasers is 20-30 sec. Disturbing foci in scars and teeth
Another important application of laser treatment is
Case Example: Perforating corneal injury in a pony elimination of disturbing foci in scars and teeth. In the
gelding author’s experience, these foci often play an essential role
A pony gelding had his eye pierced by a pitchfork in development of chronic disease.5, 11 With their discovery
tip. The cornea was completely perforated with a lesion and treatment, one can greatly enhance the healing process
diameter of 7mm. The animal was pretreated carefully of some diseases and provide a good chance for resolution.
by another vet for over 3 weeks with general and local A disturbing focus mainly arises from larger scars,
antibiotics, eye ointment and an eye bandage. Medication especially when a delayed wound healing or drainage
for pain reduction included 15ml Novalgin®c solution preceded the chronic disease. Further, particularly
applied daily. Thereafter, laser acupuncture was started. persistent disturbing foci can include infected tooth roots
Ophthalmic examination revealed a very painful and light- or non-painful inflammatory processes which are difficult
sensitive eye with a deep corneal ulcer. The cornea was to detect. In the treatment of all these disturbance foci,
opaque, swollen, had a purulent discharge and a distinct laser acupuncture offers impressive opportunities. The
reddish/yellow-red purulent exudate in the anterior local therapy is done with Nogier Fr A’’ for 1 - 3 minutes
chamber (Figure 14). per localization (15-60 joules). Beside Fr A’’ we have a
The following points were treated with the impulse second kind of special teeth disturbing foci which are
lasera for 30 sec each with Fr A’ and A’’: ST-1, GB-1, BL-1, treated by fr. Bahr 7 (38,386 Hz.) An effective acupuncture
Figure 14 and 15: Panophtalmia in pony gelding after perforation of cornea (pitchfork-injury) and conventional therapy for
3 weeks, before laser acupuncture (left photo) and after 18 days of laser acupuncture (right photo).

method to support disturbing focus treatment is the 5. Petermann, U. The Components of the Pulse
acupoint TH-5, an important immunologically effective Controlled Laser Acupuncture, PCLAC, AJTCVM
point that promotes the healing of granulation tissue in 2012; 7 (1): 57-67.
the focal disturbance. A very pleasant side effect is that 6. Mester E. Jassagi-Nagy, E. Experimentelle
the scars almost completely regress after a few treatments. Untersuchungen über die Wirkung von Laserstrahlen
auf die Wundheilung. Z. Exper. Chirurgie 1969:
Conclusion 2,94-101.
Acupuncture and laser therapy are two separate 7. Karu T. et al. Suppression of human blood
therapeutic modalities that when combined have a unique chemiluminescence by
synergism. In the author’s 28 year experience with several 8. diode laser irradiation. Laser Therapy 5, 1993: 103-
thousand animals, the combination of these therapies 109.
as laser acupuncture is an extremely effective treatment 9. Petermann, U. (2015) Wound Infection Combination
option for almost all diseases found in horses and dogs. of Laser Acupuncture and Low Level Laser Therapy
With the use of suitable impulse lasers, this therapy is for Treatment of Non-healing and Infected Wounds,
capable of exerting profound anti-inflammatory and AJTCVM 2015; 10 (2): 33-42.
antibacterial effects along with the ability to promote 10. Skobelkin O.K. et al. Blood Microcirculation under
healing in devitalized tissue. The local effect of the laser Laser Physio-and Reflexotherapy in Patients with
with energy supplied in the form of ATP is excellently lesions in Vessels of Low Extremeties. Laser Therapy
supplemented by an autonomous regulation of the immune 1990: 69-77.
system and stimulation of organ functions by acupuncture. 11. Petermann, U. Comparision of Pre- and Post-
It is hoped that more veterinarians, particularly those with treatment Pain Scores of Twenty One Horse with
acupuncture expertise, will use this truly versatile therapy. Laminitis Treated with Acupoint and Topical Low
It provides another tool in the clinician’s tool box and in Level Impulse Laser Therapy, AJTCVM 2011; 6 (1):
the author’s experience is able to rescue cases that were 13-25.
thought to be hopeless. This “holistic treatment” not 12. Petermann, U. Pulse Controlled Laser Acupuncture
only treats a focal problem in the back, hip or tendon but Concept (PCLAC), Textbook for controlled
endeavors to treat the whole animal. acupuncture, 2007; self-published, www.
akupunkturtierarzt.de.
FOOTNOTES
13. Nogier, P. F.M. Lehrbuch der Auriculotherapie
a. The Physiolaser Olympic; 90 W/904nm, super pulsed
Maisonneuve; 1981, Sainte Ruffine.
laser. Reimers & Jansenn, 14057, Berlin, Germany
14. Bahr, F., Strittmatter, B. Das große Buch der
b. Laserpen; 70 mW/785nm, continuous wave laser.
Ohrakupunktur, 2010; Hippokrates Verlag, Stuttgart.
Reimers & Jansenn, 14057, Berlin, Germany
15. Reininger, M. Die meridianspezifischen Laser-

.c Novalgin® Sanofi-Aventis Deutschland GmbH
Frequenzen nach Reiniger EAA Expertenkongress,
Industriepark Höchst / Gebäude K607
Der Akupunkturarzt 4; 2002: 11-15.
65926 Frankfurt am Main
16. Petermann, U. Treatment of 29 Cases of Acute and
Chronic Equine Tendonitis with Local Laser Therapy
REFERENCES
and Laser Acupuncture, AJTCVM 2016; 11, (2): 43-
1. Hamblin MR, Demidova TN. Mechanisms of low
51.
level light therapy, Proc. SPIE. 2006: 6140, doi:
17. Bergsmann, O. Die biokybernetische Wirkung der
10.1117/12.646294.
Akupunktur im klinischen Versuc, Dtsch. Ztschr. f.
2. Karu TI. Photobiological Fundamentals of Low-
Akup. 1977; 5 :131ff.
Power Laser Therapy. IEEE Journal od Quantum
18. Petermann, U. Behandlung von BWS- und LWS-
Electronics QE-23, 1987: 1703-1717.
Beschwerden beim Pferd mit Ohrakupunktur.
3. Warnke, U. Der Dioden-Laser, Deutsches Ärzteblatt,
collegium veterinarium; 1989: 20/91-93.
44, 1987: 2941-2944
19. Petermann, U. Laser Acupuncture in the Treatment
4. Warnke, U. Wie Licht-Energie zu Zell-Energie wird.
of Chronic Back Pain in Horses, Procc. of 28th IVAS
Ärztliche Praxis Jahrg. 97, 1987: 3039-3040.
2001 World Congress, Liuhe Hawaii, USA: 81-88.

PCLAC book Dr. Uwe Petermann DVM Topics of the PCLAC Book
28 Years Expertise in Laser Acupuncture
 RAC Pulse Control to find „active“ acupoints

Pulse Controlled Laser Acupuncture Concept
 Laser Acupuncture for painless and forceful acupoint stimulation
 Auricular Medicine/Ear Acupuncture for diagnosis and treatment
 Low Level Laser Therapy on top of acupuncture for local tissue
This book describes the holis�c modern
acupuncture treatment concept for nearly healing, infected wounds, tendons, spinal cord- nerve lesions
all thinkable kind of disease with all the  Perturbative field treatment by topical laser treatment of teeth
tradi�onal and modern scien�fic and scars which are the main causes in chronic degenerative,
chronic allergic and auto immune disease
backgrounds in theory and by case studies.
Hardcover 280 pages, 200 photos and

graphics.
ADVANCED TOPIC COURSE on
Canine and Equine Auricular Medicine/
About the book and how to purchase:
Ear Acupuncture/Gold Bead Implants into Ear
Points. State of Art in Vet. Auricular Medicine
www.akupunkturtierarzt.de Dr. Uwe Petermann, Germany
Link „English Home“ May 5-7 2017 Fort Collins, Colorado
ABSTRACT
Wang Y1, Zhang Y, Wang W, Cao Y, Han JS.

Effects of synchronous or asynchronous electroacupuncture stimulation with low versus high frequency on spinal
opioid release and tail flick nociception.
Exp Neurol. 2005 Mar;192(1):156-62.
Electroacupuncture stimulation (EAS) is known to change brain neurotransmitter release. In the present study, we
investigated the effects of synchronous or asynchronous electroacupuncture stimulation with low versus high frequency
on spinal opioid release and tail flick nociception. Rats were given “2/100 Hz” EAS, which stands for an asynchronous
mode of stimulation, in which 2 Hz was alternated with 100 Hz, each lasting for 3 s, or “(2 + 100) Hz” EAS, a mode of
stimulation in which 2 Hz stimulation was applied to the left hind leg simultaneously with 100 Hz stimulation on the right
hind leg. The rats were subjected to the same total number of electrical stimulations in these two modes. Results were as
follows: (1) 2/100 Hz EAS was 40% more potent than (2 + 100) Hz EAS (P < 0.01) in producing an anti-nociceptive effect.
(2) Intrathecal (i.t.) injection of the mu-opioid receptor antagonist D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr amide (CTOP)
blocked in a dose-dependent manner the anti-nociceptive effect produced by 2/100 Hz EAS but not by (2 + 100) Hz EAS,
whereas i.t. injection of the kappa-opioid receptor antagonist norbinaltorphimide (Nor-BNI) blocked the anti-nociceptive
effect induced by both modes of EAS. (3) I.t. injection of endomorphin-2 antiserum blocked in a dose-dependent manner
the anti-nociceptive effect of 2/100 Hz EAS but not that of (2 + 100) Hz EAS, whereas i.t. injection of dynorphin antiserum
blocked the anti-nociceptive effect induced by both modes of stimulation. (4) 2/100 Hz EAS increased the release of both
endomorphin-2 and dynorphin, whereas (2 + 100) Hz EAS increased the release of dynorphin but not of endmorphin-2.
We conclude that the more potent anti-nociceptive effect induced by 2/100 Hz EAS, as compared with that of (2 + 100) Hz
EAS, was due, at least partly, to the synergistic interaction of endomorphin-2 and dynorphin in rat spinal cord.

Gui Zhi (Cinnamomum)

Common Name: Cinnamon twig
Botanical Name: Cinnamomum cassia Presl
Part Used: Twig
Channel/Organ: Heart, Lung, Bladder
Taste: Pungent, Sweet
Energy: Warm
Energetic Functions:
1. Promote sweating and clear the surface
2. Promote Blood circulation
3. Warm the Channels and dispel Cold
4. Relieve surface pain, especially in the upper body
5. Regulate the Ying and Wei
Comments:
1. Combining with Ma Huang (Ephedra) to dispel Wind-Cold invasion
2. Contraindication for Yin Deficiency with Heat, Blood Heat or Excessive Fire
3. Caution during pregnancy
4. Overdose or long-term use may cause dryness of the throat and mouth
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Herbal Formula Spotlight
Tao Hong Si Wu Tang (Four-Substance Decoction with Safflower and Peach Pit)
Tao Hong Si Wu Tang (Four-Substance Decoction with Safflower and Peach Pit) contains Si Wu Tang (Four-
Substance Decoction) plus Tao Ren (Semen Persicae) and Hong Hua (Flos Carthami). Si Wu Tang is considered to be the
representative formula for tonifying the blood. The inclusion of the herbs Tao Ren and Hong Hua enables this formula
to dispel blood stagnation in addition to tonifying the blood and regulating blood circulation. The original source of the
formula is from the Yi Zong Jin Jian (Golden Mirror of the Medical Tradition) by Wu Qian in 1742 .1 The original formula
of Tao Hong Si Wu Tang is composed of the following individual herbs: 12g Shu Di Huang (Rehmannia), 9g Dang Gui
(Angelica), 9g Bai Shao (Paeonia), and 6g Chuan Xiong (Ligusticum), 9 g Tao Ren (Semen Persicae), 3 g Hong Hua
(Flos Carthami) .2 The Chinese therapeutic actions of this formula are to nourish blood, regulate blood circulation and
dispel blood stasis. All of the herbs in the formula Si Wu Tang enter the blood level. Shu Di Huang (Radix Rehmanniae
Praeparata), the chief herb, tonifies the blood. It also tonifies the Kidney, which generates the marrow necessary for
blood production. When tonifying blood, it is important to also move the blood to prevent stagnation. Dang Gui (Radix
Angelicae Sinensis) tonifies the blood, activates blood circulation, and nourishes the Liver. Bai Shao (Radix Paeoniae
Alba) nourishes the blood and consolidates yin. It harmonizes the blood in the ying (nutritive) level. Chuan Xiong
(Rhizoma Chuanxiong) activates and regulates blood circulation, and prevents the tonic herbs from causing stagnation.
The addition of the herbs Tao Ren (Semen Persicae) and Hong Hua (Flos Carthami) act to regulate and invigorate blood
circulation and eliminate blood stagnation. Clinically Tao Hong Si Wu Tang has been used in veterinary medicine to
treat infertility, irregular estrus cycles in breeding animals, lumbar pain occurring at estrus, diabetic neuropathy, non-
healing bone fractures, progressive retinal atrophy, chronic superficial keratitis (CSK), non-healing corneal ulcer, optic
neuritis, cataracts, glaucoma, urticaria, itching, atopic dermatitis and seborrhea. Animals with blood deficiency and
blood stagnation often present with a pale purple tongue and a weak, wiry or choppy pulse. Tao Hong Si Wu Tang is
designed to treat disorders characterized by blood deficiency with blood stagnation. To potentiate its effects, Bai Shao
(Radix Paeoniae Alba) can be replaced with Chi Shao (Radix Paeoniae Rubra) to enhance the blood-activating effect,
and Shu Di Huang (Radix Rehmanniae Praeparata) can be replaced with Di Huang (Radix Rehmanniae) to nourish the
blood without creating more stagnation. Since Tao Hong Si Wu Tang has strong actions to regulate blood circulation and
eliminate blood stasis, it should be discontinued once the desired effects are achieved and should be used with caution
1
in animals with underlying qi and blood deficiencies. It should be kept in mind, that because it moves the blood, it
could potentially cause excessive bleeding in some animals if used immediately prior to a surgical procedure. It is
recommended to discontinue this formula at least 5 days prior to any surgical procedure.
Horses and Cattle: 15–60g, twice daily. Camels: 30–75g, twice daily. Llamas, Alpacas, Pigs, Goats, and Sheep:
5–15g, twice daily. Dogs: 0.5–5g (or 0.1g per kg body weight), twice daily. Cats and Rabbits: 0.2–0.5 g (or 0.1 g per kg
body weight), twice daily. Birds: 0.1–0.2g per kg body weight, twice daily.2
Tao Hong Si Wu Tang is contraindicated during pregnancy.
The pharmacological effects of Tao Hong Si Wu Tang include cardiovascular, anti-hyperlipidemic and anti-
inflammatory effects. An in vitro study in human patients with coronary artery disease showed that Tao Hong Si Wu
3
Tang can dilate the coronary artery and increase blood perfusion to cardiac muscles. Administration of Tao Hong Si Wu
4
Tang in rats was shown to have anti-hyperlipidemic effects by decreasing both total cholesterol and triglyceride levels.
According to another in vitro study in rats, Tao Hong Si Wu Tang was associated with marked anti-inflammatory effects
5
similar to hydrocortisone, and did not produce any of the side effects associated with hydrocortisone administration.
Human clinical studies showed that 32 patients with cerebral infarction were treated with complete recovery in 10
patients, significant improvement in 11 patients, moderate improvement in 9 patients, and no benefit in 2 patients. The
herbal treatment contained Tao Hong Si Wu Tang plus Ge Gen (Radix Puerariae Lobatae), Shui Zhi (Hirudo) and Huang
6
Qi (Radix Astragali). In another study modified Tao Hong Si Wu Tang was effective in treating female infertility caused
by blocked fallopian tubes. The base herbal treatment consisted of Tao Hong Si Wu Tang plus Xiang Fu (Rhizoma
Cyperi), Shui Zhi (Hirudo), Lu Lu Tong (Fructus Liquidambaris), Dan Shen (Radix et Rhizoma Salviae Miltiorrhizae),
7
and San Qi (Radix et Rhizoma Notoginseng) powder. The use of modified Tao Hong Si Wu Tang to treat diabetic
neuropathy: was associated with 96.4% effectiveness in treating peripheral neuropathy in 28 diabetic patients (22 with
more than 5 years of illness, and 6 with less than 5 years of illness). The base formula contained Tao Hong Si Wu
Tang plus Huang Qi (Radix Astragali), Dang Shen (Radix Codonopsis), Huang Jing (Rhizoma Polygonati) and Gou
Qi Zi (Fructus Lycii).) Of 28 patients, the study reported complete recovery in 15 patients, moderate improvement in 9
8
patients, slight improvement in 3 patients, and no benefit in 1 patient. In the treatment of bone fractures Tao Hong Si
Wu Tang was combined with Gu Sui Bu (Rhizoma Drynariae), Xu Duan (Radix Dipsaci), Zi Ran Tong (Pyritum), Tu Bie

Chong (Eupolyphaga seu Steleophaga), Mu Xiang (Radix Aucklandiae), and others as needed. Additional modifications
included Huang Bo (Cortex Phellodendri Chinensis) for heat; unprocessed Cao Wu (Radix Aconiti Kusnezoffii) for cold;
Sang Zhi (Ramulus Mori) for bone fractures in the upper extremities; and Niu Xi (Radix Achyranthis Bidentatae) for
9
bone fractures in the lower extremities. Use of modified Tao Hong Si Wu Tang was shown to be effective in treating
allergic rhinitis in 42 patients. The base formula contained Tao Hong Si Wu Tang plus Cang Er Zi (Fructus Xanthii),
10
Huang Qi (Radix Astragali), Fang Feng (Radix Saposhnikoviae), and Xin Yi Hua (Flos Magnoliae). One study reported
93.5% effectiveness using modified Tao Hong Si Wu Tang to treat itching. It was combined with He Shou Wu (Radix
Polygoni Multiflori) to nourish blood and Di Fu Zi (Fructus Kochiae), Bai Xian Pi (Cortex Dictamni), and Ji Li (Fructus
Tribuli) to control pruritis. Of 92 patients, the study reported complete recovery in 53 patients, moderate improvement
11
in 33 patients, and no benefit in 6 patients. Administration of Tao Hong Si Wu Tang plus Dan Shen (Radix et Rhizoma
Salviae Miltiorrhizae), Chi Shao (Radix Paeoniae Rubra), and Mu Dan Pi (Cortex Moutan) in 64 patients with eczema
was associated with complete recovery in 53 patients, significant improvement in 5 patients, and slight improvement
12
in 6 patients. One study reported beneficial effect in 32 patients with optic neuritis using Tao Hong Si Wu Tang plus
Dan Shen (Radix et Rhizoma Salviae Miltiorrhizae), Zhi Qiao (Fructus Aurantii), Fu Ling (Poria), Tong Cao (Medulla
13
Tetrapanacis), and Gan Cao (Radix et Rhizoma Glycyrrhizae).
Signe E Beebe DVM
Table 1: The ingredients of the Chinese herbal medicine Tao Hong Si Wu Tang (Four-Substance Decoction with Safflower
and Peach Pit) and their actions1,2
Pin Yin Name English Name Amount (g) Actions
Shu Di Huang Rehmannia 6g Tonifies yin and promotes blood production
Dang Gui Angelica 6g Tonifies the blood, activates blood circulation
Bai Shao White Peony 6g Nourishes the blood, consolidates yin, and harmonizes the blood
Chuan Xiong Ligusticum 3g Activates and regulates blood circulation
Tao Ren Persica 6g Activate Blood, dispel stasis
Hong Hua Cathamus 3g Activate Blood, dispel stasis
REFERENCES
1. Beebe, S. Salewski, M, Chen J, Chen T. Chinese Herbal Formulas for Veterinarians. City of Industry, CA., Art
of Medicine Press: 2012:569-573.
2. Xie H, Preast V. Xie’s Chinese Veterinary Herbology. Ames, IA:Wiley-Blackwell 2010: 454.
3. Jiang Su Zhong Yi Za Zhi (Jiangsu Journal of Chinese Medicine), 1986; 12:541.
4. Zhong Xi Yi Jie He Za Zhi (Journal of Integrated Chinese and Western Medicine), 1988; 8(10):601.
5. Zhong Guo Zhong Yao Za Zhi (People’s Republic of China Journal of Chinese Herbology), 1994; 19(11):680.
6. He Bei Zhong Yi (Hebei Chinese Medicine), 1995; 15(2):26.
7. Zhong Yi Han Shou Tong Xun (Reports of Chinese Medicine), 1992; 3:44.
8. Shi Yong Zhong Yi Nei Ke Za Zhi (Journal of Practical Chinese Internal Medicine), 1991; 5(2):73.
9. Zhong Yi Ming Fang Lin Chuang Xin Yong (Contemporary Clinical Applications of Classic Chinese Formulas),
2001; 1:29.
10. Yan Ke Tong Xun (Journal of Ophthalmology), 1987; 3:13.
11. Shi Yong Zhong Xi Yi Jie He Za Zhi (Practical Journal of Integrated Chinese and Western Medicines), 1991; 4:41.
12. Zhong Yi Za Zhi (Journal of Chinese Medicine), 1988; 29(5):366.
13. ZhongYiFangJiXianDaiYanJiu (ModernStudy of Medical Formulas in Traditional Chinese Medicine), 1997; 1103.

WATCVM
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All food supports the Earth Element
To prevent damage to the Spleen avoid over eating/feeding foods that are Dampening. Eat/feed in moderation
with the goal to keep the patient trim. Avoid excessive amounts of Cold foods. Cold damages the Middle Jiao
so the Spleen/Stomach cannot ripen food for proper transformation or transportation. Earth foods support
other organ systems and the five treasures. The goals are to tonify organs and treasures and balance according
to TCVM temperatures. Variety is important!
Foods which support the Earth, as well as other organ systems.
Meat/Protein Vegetable Fruit Grain/legume
Wood Beef, egg, mutton Mushrooms, spinach Apple, cherry Kidney bean, oats
Fire Beef, egg, lamb Pumpkin, tomato Apple, cherry Kidney bean, oats
Metal Egg, mutton Mushrooms, pumpkin, yam Apple, cherry Navy beans (great northern), oats
Water Beef, egg, lamb Pumpkin, yam Apple, cherry Kidney bean, oats
Earth Balancing Stew Recipe

2 cups Beef, diced to the size appropriate for the pet
1 tbs Olive oil for browning beef
Spinach, fresh (1/4 cup for cats, 1 cup for dogs)
Mushrooms, sliced, raw (1/8 cup for cats, ½ cup for dogs)
Kidney beans, cooked, canned (omit for cats, 2 cups for dogs)
Yam, baked or boiled (1/8 cup for cats, 1 cup for dogs)
Hardboiled egg (2 tsp for cats, whole for dogs)
Lightly brown beef in a little olive oil. Quickly

sauté spinach and mushrooms in oil at end
of cooking for the beef. Mix in kidney
beans, baked yam and hardboiled
egg when cooking is complete.
Sliced or dehydrated apples
for snacks.
Back cover content and design courtesy of Dr. Cathy Alinovi DVM, CVSMT, CTCVMP

American Journal of Traditional Chinese Veterinary Medicine

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AMERICAN JOURNAL OF

TRADITIONAL CHINESE VETERINARY MEDICINE

VOLUME 12, NUMBER 1 February 2017

The Official Journal of the American Association of

Join Your Veterinary Colleagues

Member Benefits: Mission:

• REGIONAL CE WITH DISCOUNTED REGISTRATION

AJTCVM Ronald Koh DVM

AJTCVM Vol. 12, No. 1, February 2017 i

About the Cover

ii AJTCVM Vol. 12, No. 1, February 2017

The American Journal of Traditional Chinese

February 2017 Vol. 12 No. 1

TCVM News and Educational Opportunities.................................................................................................................................5

AATCVM Updates, Letter from the President.............................................................................................................................9

IVAS Updates, Letter from the President......................................................................................................................................13

WATCVM Updates, Letter from the President.............................................................................................................................17

Basic Science Studies

A Randomized and Controlled Experimental Study on the Effects of Epimedium-Propolis Adjuvant

AJTCVM Vol. 12, No. 1, February 2017 iii

Pearls from TCVM Practice

Acupuncture and endorphins.........................................................................................................................................................8

Herbal Materia Medica

Chinese Herbal Medicine Spot Light

iv AJTCVM Vol. 12, No. 1, February 2017

Chi Institute International TCVM Conference................................................................................................................78

Instructions to Authors – visit www.ajtcvm.org (authors tab)

AJTCVM Vol. 12, No. 1, February 2017 v

Black Box Theory and Diagnostic System of

Huisheng Xie DVM, MS, PhD

History of Black Box functions internally by the relationship between the

Table 1: The Four Diagnostic Methods (Si-Zhen)

• Examine the tongue characteristics and color patterns

• Listen to the voice quality (weak, muffled, strong and/or robust)

2 AJTCVM Vol. 12, No. 1, February 2017

Figure 3: The Black Box approach of the TCVM Diagnosis

4 AJTCVM Vol. 12, No. 1, February 2017

TCVM Around the World and Educational

AJTCVM Vol. 12, No. 1, February 2017 5

spinal manipulative therapy, veterinary massage and

Biography of Dr. Caili

Chinese Herbs & Herbal products since 1969

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I am Keum-Hwa Choi. I have been elected as clinician.

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Upcoming AAVA Events

FAAVA Exam: Want to Take the Next Step in your

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Bruce Ferguson DVM, MS President AATCVM

Dr. Ferguson with Suzanne Hughes and Sunny at Upper

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2015-2017 AATCVM Funded Research Grants

Effect of oral administration of Yunnan Baiyao on

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Improve your ophthalmic

diagnostic and surgical skills

Earn IVAS CEUs Online

Learn from world-class speakers on topics including:

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Dr. Uwe Petermann

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Group photo of speakers. Gene Tagaban demonstrating the Raven Dance.

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