This work was supported by National Natural Science Foundation of China (No. 81922076, 81973951, 81873373), Construction project of collaborative innovation of shanghai (No. ZYJKFW201811010, ZYJKFW201701005).

All animal experiments were reviewed and approved by the Committee on the Ethics of Animal Experiments of Shanghai University of Traditional Chinese Medicine (Ethical clearance number PZSHUTCM190308020).
1. Fabrication of the high platform
<ol>
	<li>Set the length and width of the supporting device to 86 cm x 17 cm (Figure 1).</li>
	<li>Fix the connecting device vertically to the supporting device. The connecting device should be approximately 30 cm long to prevent the rat from running to the top surface of the high platform and have a width of 14 cm.</li>
	<li>Fix the horizontal pedal (5 cm x 5 cm) to the connecting device. The suspended height at the bottom of the horizontal pedal should be greater than or equal to 50 cm. 
	NOTE: Depending on the number of experimental rats, multiple horizontal pedals can be used, with one rat placed on each pedal.</li>
</ol>
2. Establishment of a rat model of asthma
<ol>
	<li>Sensitizing the rats with OVA
	<ol>
		<li>Prepare the sensitization solution on day 0. Prepare each 1 mL of the sensitization solution with 1 mg of OVA and 10 mg of aluminum hydroxide, with normal saline as the solvent.</li>
		<li>Intraperitoneally inject each rat with 1 mL of the sensitization solution on day 0. Draw the sensitizing solution with a suitable syringe for later use.</li>
		<li>Grab the tail and lift up the rat, place it on a rough surface, and gently pull the tail back. Grasp the skin of both ears and neck with the left thumb and index finger while tightly holding the back skin with the remaining three fingers and palms (be careful to refrain from applying excess force that will suffocate the rat).</li>
		<li>With the right hand, perform an intraperitoneal injection in the rat with the prepared syringe with 1 mL of the sensitization solution.</li>
	</ol>
	</li>
	<li>Challenging rats with OVA
	<ol>
		<li>Select a box with a lid (the length, width and height are 39 cm x 28 cm x 23 cm), and drill a small hole in the side. Connect the sealed box, nebulizing cup and the compression nebulizer with a rubber tube (Supplementary Figure 1). 
		NOTE: The nebulized particle size of the compressed nebulizer is 1 - 5 &#956;m, the nebulization rate is greater than 0.2 mL/min, and the ultimate pressure is 205 kPa.</li>
		<li>On the 14th day after sensitization, open the lid of the homemade sealed box and place the rat in the box (the box can accommodate 5 rats), and then put the sealed box in a biosafety cabinet.</li>
		<li>Prepare the challenge solution. Thoroughly mix 300 mg of OVA with 30 mL of normal saline.</li>
		<li>Turn on the ultrasonic nebulizer and add 30 mL of the challenge solution into the nebulizing cup. Adjust the dosage of challenge solution and the number of rats according to the volume of the homemade sealed box.</li>
		<li>Repeat nebulization several times (once a day) or adjust the procedure according to the severity of asthma symptoms.</li>
	</ol>
	</li>
</ol>
3. Acupuncture treatment
<ol>
	<li>Select acupoints, including GV14, bilateral BL12 and bilateral BL13, according to the theory of traditional Chinese medicine for treating asthma (Supplementary Figure 2). Administer manual acupuncture on day 2 and perform once every other day for a total of 7 session (Figure 2).
	<ol>
		<li>Acupuncture the GV14 located between the seventh cervical vertebra and the first thoracic vertebra (the hollow of the scapula), on the midline of the back (Figure 3).</li>
		<li>Acupuncture the BL13 located below the third thoracic vertebra, on both sides of the intercostal spaces (lower edge of the scapula).</li>
		<li>Acupuncture the BL12 located below the second thoracic vertebra, on both sides of the intercostal spaces (below the scapula, position identified according to BL13).</li>
	</ol>
	</li>
	<li>Grasp the rats and place them on the high platform.</li>
	<li>Hold the disposable stainless-steel needle (0.3 mm x&#160;13.0 mm, Supplementary Figure 3) handle with the thumb and index finger of the right hand, and press the back of the rat with the palm of the right hand. 
	NOTE: Disposable needles must be one-time-use only. Because repeated use will blunt the needle, which leads to increased pain in rats.</li>
	<li>Touch the depression in the middle of the upper edge (GV14) or the lower edge of the scapula (BL12 and BL13), and then gently prick the skin of the rat with the needle tip to locate the acupoints.</li>
	<li>Quickly insert the needle after determining the specific location of the acupoint and rapidly remove the right hand.</li>
	<li>Evenly twisted the needles approximately 360&#176; at a rate of 60 times/min for 10 - 20 s, manipulate them every 5 min, and then withdraw them after 20 - 30 min (Supplementary Figure 4).</li>
</ol>
4. The Penh measurement
<ol>
	<li>Prepare methacholine: Place 100 mg of methacholine into a 5 mL centrifuge tube and add PBS to a volume of 2 mL (50 mg/mL methacholine). Then, dilute the 50 mg/mL methacholine solution to 25 mg/mL, 12.5 mg/mL, 6.25 mg/mL and 3.125 mg/mL.</li>
	<li>Instrument inspection
	<ol>
		<li>Turn on the signal amplifier (the aerosol distribution system and the rodent bias flow supply system are all turned off, and the nebulization connector is closed).</li>
		<li>Check the channels: Click Calibrate, return the coarse adjustment baseline to 0 and switch to the direct current (DC) position. Fine-tune the baseline to 0 and inject 5 mL of gas with a syringe. Then, switch to the alternating current (AC) position.</li>
		<li>After checking the channels, ensure that the error value is less than 0.5%.</li>
	</ol>
	</li>
	<li>Setting parameters
	<ol>
		<li>Turn on the aerosol distribution system and the rodent bias flow supply system. Adjust the flow rate to 4.5 mL according to the liquid level on the mercury ball.</li>
		<li>Click Create Study Options and select Dose Response Study, select Rat of species and click Next.</li>
		<li>Select Simple Dose List, set the Number of Doses to 6 and click Next.</li>
		<li>Select the Penh and Area parameters: Set the animal adaptation time to 5 min, the nebulization time to 2 min, the response time to 3 min, and the recovery time to 2 min.</li>
		<li>Sequentially add the following solutions to the nebulizer (200 &#956;L per rat): methacholine (0 mg/mL, 3.125 mg/mL, 6.25 mg/mL, 12.5 mg/mL, 25 mg/mL and 50 mg/mL). Real-time changes in Penh can be observed (Figure 4).</li>
		<li>After all 6 solutions with different concentrations have been reacted, click File and End Session to complete the experiment.</li>
	</ol>
	</li>
</ol>

<ol>
	<li>Papi, A., Brightling, C., Pedersen, S. E., Reddel, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asthma.">Asthma.</a> Lancet. 391 (10122), 783-800 (2018).</li><li>. Global strategy for asthma management and prevention Available from: <a target="_blank" href="https://ginasthma.org/gina-reports/">https://ginasthma.org/gina-reports/</a> (2019)</li><li>Enilari, O., Sinha, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Global+Impact+of+Asthma+in+Adult+Populations.">The Global Impact of Asthma in Adult Populations.</a> Annals of Global Health. 85 (1), (2019).</li><li>. Global strategy for asthma management and prevention Available from: <a target="_blank" href="https://ginasthma.org/gina-reports/">https://ginasthma.org/gina-reports/</a> (2020)</li><li>Zhuang, Y., Xing, J. J., Li, J., Zeng, B. Y., Liang, F. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=History+of+acupuncture+research.">History of acupuncture research.</a> International Review of Neurobiology. 111, 1-23 (2013).</li><li>Yang, Y. Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Considerations+for+use+of+acupuncture+as+supplemental+therapy+for+patients+with+allergic+asthma.">Considerations for use of acupuncture as supplemental therapy for patients with allergic asthma.</a> Clinical Reviews in Allergy &amp; Immunology. 44 (3), 254-261 (2013).</li><li>Nurwati, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+interleukin-17+level+by+acupuncture+at+Feishu+(BL13)+is+strengthened+by+acupuncture+at+Zusanli+(ST36)+in+a+mouse+model+of+chronic+asthma:+An+experimental+study.">Reduction of interleukin-17 level by acupuncture at Feishu (BL13) is strengthened by acupuncture at Zusanli (ST36) in a mouse model of chronic asthma: An experimental study.</a> Medical Acupuncture. 27 (4), 278-282 (2015).</li><li>Nurwati, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvement+in+inflammation+and+airway+remodelling+after+acupuncture+at+BL13+and+ST36+in+a+mouse+model+of+chronic+asthma.">Improvement in inflammation and airway remodelling after acupuncture at BL13 and ST36 in a mouse model of chronic asthma.</a> Acupuncture in Medicine. 37 (4), 228-236 (2019).</li><li>Reddy, A. T., Lakshmi, S. P., Reddy, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+model+of+allergen+induced+asthma.">Murine model of allergen induced asthma.</a> Journal of Visualized Experiments. (63), e3771 (2012).</li><li>Zhou, D. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metallothionein-2+is+associated+with+the+amelioration+of+asthmatic+pulmonary+function+by+acupuncture+through+protein+phosphorylation.">Metallothionein-2 is associated with the amelioration of asthmatic pulmonary function by acupuncture through protein phosphorylation.</a> Biomedicine &amp; Pharmacotherapy. 123, 109785 (2020).</li><li>Yin, L. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgelin-2+as+a+therapeutic+target+for+asthmatic+pulmonary+resistance.">Transgelin-2 as a therapeutic target for asthmatic pulmonary resistance.</a> Science Translational Medicine. 10 (427), (2018).</li><li>Shin, Y. S., Takeda, K., Gelfand, E. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+asthma+using+animal+models.">Understanding asthma using animal models.</a> Allergy Asthma &amp; Immunology Research. 1 (1), 10-18 (2009).</li><li>Cui, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electro-acupuncture+relieves+chronic+visceral+hyperalgesia+in+rats.">Electro-acupuncture relieves chronic visceral hyperalgesia in rats.</a> Neuroscience Letters. 376 (1), 20-23 (2005).</li><li>Tobin, J. R., Martin, L. D., Breslow, M. J., Traystman, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+anesthetic+inhibition+of+brain+nitric+oxide+synthase.">Selective anesthetic inhibition of brain nitric oxide synthase.</a> Anesthesiology. 81 (5), 1264-1269 (1994).</li><li>Dimitrov, N., Sivrev, D., Atanasova, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Method+of+rat+immobilization+during+experimental+acupuncture.">Method of rat immobilization during experimental acupuncture.</a> Scripta Scientifica Medica. 45, 105-108 (2013).</li><li>Aguilar, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=emotional+reactivity+and+fear+of+heights:+a+factor+analytic+map+from+a+large+F(2)+intercross+of+Roman+rat+strains.">emotional reactivity and fear of heights: a factor analytic map from a large F(2) intercross of Roman rat strains.</a> Brain Research Bulletin. 57 (2), 17-26 (2002).</li><li>Lai, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteomic+response+to+acupuncture+treatment+in+spontaneously+hypertensive+rats.">Proteomic response to acupuncture treatment in spontaneously hypertensive rats.</a> PloS One. 7 (9), 44216 (2012).</li><li>Niu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+uni-acupoint+electroacupuncture+stimulation+method+for+pain+relief.">A novel uni-acupoint electroacupuncture stimulation method for pain relief.</a> Evidence-Based Complementary and Alternative Medicine. 2011, 209879 (2011).</li><li>McFadden, E. R., Luparello, T., Lyons, H. A., Bleecker, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mechanism+of+action+of+suggestion+in+the+induction+of+acute+asthma+attacks.">The mechanism of action of suggestion in the induction of acute asthma attacks.</a> Psychosomatic Medicine. 31 (2), 134-143 (1969).</li><li>Savov, J. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ozone-induced+acute+pulmonary+injury+in+inbred+mouse+strains.">Ozone-induced acute pulmonary injury in inbred mouse strains.</a> American Journal of Respiratory Cell and Molecular Biology. 31 (1), 69-77 (2004).</li></ol>

The authors have nothing to disclose.

The GV14, bilateral BL12 and bilateral BL13 acupoints used in this study are located on the back, and thus the high platform is suitable for fixing rats. In other studies, an adequate fixation method should be selected according to the position of the acupoints15. For acupoints on the abdomen, the rat can be placed downward in a black cone (similar to a pastry bag used by bakers)17. A chain is attached to the restraint cone for fixation, and the hind leg of the rat is also ...

Asthma is a chronic disease characterized by bronchoconstriction and airway hyper-responsiveness1. Approximately 339.4 million people worldwide are diagnosed with asthma2. The prevalence of asthma is increasing globally, with 100 million new cases expected in the next decade3. Inhaled corticosteroids and long-acting &#946;2-agonists do not effectively control the symptoms of all patients4. Therefore, the use of alternative therapies such as acupuncture to treat asthma has been receiving increased attention5. An effective method for treating asthma is to needle GV14 (Dazhui), bilateral BL12 (Fengmen) and bilateral BL13 (Feishu)6. Acupuncture significantly reduces the levels of secretory immunoglobulin A (SIgA) in the saliva and nasal secretions of patients with allergic asthma and reduces peripheral blood eosinophil counts with a total effective rate of 85%6. Acupuncture at BL13 and ST36 (Zusanli) in mice with chronic asthma decreases the level of IL-17 by 40% and decreases the smooth muscle thickness by 33% compared with that of the ovalbumin (OVA) group, and thus relieving inflammation symptoms7,8. However, the mechanism of acupuncture treatment for asthma is not completely clear.
Animal models are an important tool for asthma research9 because the animal model can be used for the continuous measurement of respiratory function to assess the effectiveness of the asthma treatment10. Meanwhile, studies of the pathophysiology of asthma require test samples, including samples from the trachea, lung and bronchoalveolar lavage to verify changes in the levels of key factors11. Rat models of asthma are commonly used to assess asthma pathophysiology because they produce long-lasting airway responses and show immediate and late-phase airway responses12. However, due to repeated experimental stimulation, the rats often become irritable. Therefore, a suitable method to fix rats is needed. Commonly used methods for rat fixation include anesthesia and binding13. Although the anesthesia method provides better exposure of the acupoints, it may affect nerve conduction and ultimately affect the experimental results14. The binding method does not induce the same physiological effects as anesthesia, but restrained rats will experience skeletal muscle tension. Also, the needle cannot easily reach the designated position15. Here, we introduce a high platform that can be used to fix rats without restricting them and fully expose the acupoints on the back. The rats can be placed on the high platform during the administration of acupuncture. The high platform is able to accommodate the rat only while it is standing, and as it is placed a certain distance from the ground, the rats will not move because of a fear of heights16.
This article describes in detail the fabrication of the high platform, the establishment of a rat model of asthma, the operation of needling a rat on the high platform and the measurement of respiratory function using a whole-body plethysmography (WBP) system.

<table><tbody><tr><td>0.3 mm x 13.0 mm acupuncture needle</td><td>China Suzhou Medical Device Factory</td><td></td><td></td></tr><tr><td>Al(OH)&lt;sub&gt;3&lt;/sub&gt;</td><td>American Thermo</td><td>77161</td><td></td></tr><tr><td>Compression nebulizer</td><td>Jiangsu Lude Medical Electronics Co., Ltd.</td><td>NB-212C</td><td></td></tr><tr><td>Fine Pointe V2.0</td><td>American Buxco</td><td></td><td></td></tr><tr><td>Laboratory gas drying unit</td><td>American Drierite</td><td>26800</td><td></td></tr><tr><td>Methacholine</td><td>American Sigma</td><td>MKCF6054</td><td></td></tr><tr><td>Ovalbumin</td><td>American Sigma</td><td>A5503-25G</td><td></td></tr><tr><td>Phosphate Buffer solution (1x)</td><td>Gelifesciences</td><td>SH30256.01</td><td></td></tr><tr><td>Seal box</td><td>IRIS Corporation of Japan</td><td></td><td></td></tr><tr><td>Whole-body plethysmography respiratory function measurement system</td><td>American Buxco</td><td></td><td></td></tr></tbody></table>

acupuncture in a rat model of asthma

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes methods for the fabrication of the high platform, establishes a rat model of asthma and measures changes in respiratory function using a noninvasive and real-time whole-body plethysmography (WBP) system.

With acupuncture at the GV14, bilateral BL12 and bilateral BL13 points, the rat maintains a prone position. The rat can twist its head only to the outside and not the inside because one direction is blocked with a wooden board (Figure 5).
As the methacholine concentration increased, Penh shows an increasing trend and the growth rate gradually increases. At concentrations of 3.125 mg/mL, 6.25 mg/mL, 12.5 mg/mL, 25 mg/mL and 50 mg/mL, Penh increased by 18.40% &#177;...

Watch this Scientific Journal Video about Acupuncture in a Rat Model of Asthma at JoVE.com

Acupuncture in a Rat Model of Asthma

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes ...

Research

JoVE Journal

Medicine

5.9K Views.  Shanghai University of Traditional Chinese Medicine. Een hoog platform kan ratten zonder beperking repareren en de acupunten op de rug volledig blootleggen tijdens acupunctuurmanipulatie. Dit artikel beschrijft methoden voor de fabricage van het hoge platform, stelt een rattenmodel van astma vast en meet veranderingen in de ademhalingsfunctie met behulp van een niet-invasief en real-time plethysmografiesysteem voor het hele lichaam (WBP).

Video: Acupuncture in a Rat Model of Asthma

Murine Model of Allergen Induced Asthma

Asthma is a major cause of morbidity and mortality, affecting some 300 million people throughout the world.1 More than 8% of the US population has asthma, with the prevalence increasing.2 As with other diseases, animal models of allergic airway disease greatly facilitate understanding of the underlying pathophysiology, help identify potential therapeutic targets, and allow preclinical testing of possible new therapies. Models of allergic airway disease have been developed in several animal species, but murine models are particularly attractive due to the low cost, ready availability, and well-characterized immune systems of these animals.3 Availability of a variety of transgenic strains further increases the attractiveness of these models.4 Here we describe two murine models of allergic airway disease, both employing ovalbumin as the antigen. Following initial sensitization by intraperitoneal injection, one model delivers the antigen challenge by nebulization, the other by intratracheal delivery. These two models offer complementary advantages, with each mimicking the major features of human asthma.5
The major features of acute asthma include an exaggerated airway response to stimuli such as methacholine (airway hyperresponsiveness; AHR) and eosinophil-rich airway inflammation. These are also prominent effects of allergen challenge in our murine models,5,6 and we describe techniques for measuring them and thus evaluating the effects of experimental manipulation. Specifically, we describe both invasive7 and non-invasive8 techniques for measuring airway hyperresponsiveness as well as methods for assessing infiltration of inflammatory cells into the airways and the lung. Airway inflammatory cells are collected by bronchoalveolar lavage while lung histopathology is used to assess markers of inflammation throughout the organ. These techniques provide powerful tools for studying asthma in ways that would not be possible in humans.

Experimental mouse models of allergic asthma offer new possibilities for studying disease pathogenesis and developing new therapeutics. These models are well suited to measuring factors governing the allergic immune response, airway inflammation, and pulmonary pathophysiology.

Asthma is a major cause of morbidity and mortality, affecting some 300 million people throughout the world.1 More than 8% of the US population has asthma, ...

Immunology and Infection

Cognitive Improvement in Chronic Hypoxia Mice with Acupuncture

Acupuncture Treatment in a Mouse Model of Chronic Hypoxia-Induced Cognitive Dysfunction

The treatment of central nervous disorders has consistently posed significant challenges to the medical field. Acupuncture, a non-pharmacological practice rooted in traditional Chinese medicine, entails the insertion of fine needles into precise points on the body and is commonly employed for the management of diverse conditions. Recently, acupuncture has emerged as a promising therapeutic intervention for a range of neurological diseases, including anxiety and respiratory disorders. However, the potential of acupuncture in treating cognitive dysfunction induced by chronic hypoxia has not yet been explored. This paper presents a comprehensive protocol for establishing a mouse model of chronic hypoxia-induced cognitive impairment, administering mild anesthesia, performing acupuncture treatment, and assessing behavioral changes and memory abilities using open field tests and water mazes. The step-by-step protocol provides detailed instructions on accurately locating and positioning acupoints and needles for cognitive improvement. By employing this protocol, researchers can conduct systematic studies to thoroughly evaluate the therapeutic potential of acupuncture for cognitive dysfunction.

Author Spotlight: Improving Anesthesia Protocols for Enhanced Mouse Acupuncture Research

Here, we describe a protocol for implementing mild anesthesia and acupuncture treatment on a chronic hypoxia mouse model and conducting behavioral tests to assess the cognitive alterations post-treatment.

The treatment of central nervous disorders has consistently posed significant challenges to the medical field. Acupuncture, a non-pharmacological practice ...

Behavior

Assessing Tuina Therapy's Pain Relief in Chronic Constriction Rats

Characteristics of Pain Changes in Rats with Nerve Injury Within 24 hours After One-Time Tuina Intervention

Tuina, as an external treatment method of traditional Chinese medicine, has been proven to have an analgesic effect on peripheral neuropathic pain (pNP) in clinical and basic research. However, the optimal time point for the analgesic effect of tuina may vary according to different injury sensations, affecting the exploration of the initiation mechanism of tuina analgesia. 
The research used minor chronic constriction injury (minor CCI) model rats to simulate pNP and used the intelligent tuina manipulation simulator to simulate the three methods (point-pressing, plucking, and kneading) and three acupoints (Yinmen BL37, Chengshan BL57, and Yanglingquan GB34) for performing tuina therapy. The study evaluated the changes in pain within 24 h and the optimal time point for the efficacy of tuina analgesia in rats with minor CCI models by testing cold sensitivity threshold (CST), mechanical withdrawal threshold (MWT), and thermal withdrawal latency (TWL). Furthermore, the study evaluated IL-10 and TNF-&#945; expression changes through Elisa detection. The results show that tuina has both immediate and sustained analgesic effects. For the three different injury sensitivity thresholds of CST, MWT, TWL, and two cytokines of IL-10 and TNF-&#945;, the analgesic efficacy of tuina within 24 h after intervention is significantly different at different time points.

Author Spotlight: Exploring the Initiation Mechanism of 'Three-Method and Three-Acupoint' Tuina Analgesia in Rats with Nerve Injury

We present a protocol using the tuina manipulation simulator to perform the "Three-Manipulation and Three-Acupoint" tuina therapy for minor chronic constriction injury rats and evaluate the effective analgesic time points of tuina within 24 h by testing pain changes through behavioral analysis and changes in inflammatory factor expression using enzyme-linked immunosorbent assay.

Tuina, as an external treatment method of traditional Chinese medicine, has been proven to have an analgesic effect on peripheral neuropathic pain (pNP) ...

Neuroscience

A Traditional Chinese Medicine Characteristic Therapy for Bronchial Asthma: Moxibustion

Bronchial asthma is a chronic inflammatory disease of the airway which can lead to symptoms such as recurrent wheezing, shortness of breath, chest tightness, or cough, as a result of increased airway reactivity. With high diurnal variation, these symptoms often occur or worsen at night or in the morning. By burning and roasting the Chinese medica above human acupoints, moxibustion is a type of treatment that stimulates the activity of the human meridians through drugs and heat stimulation to prevent and treat diseases. According to the principle of syndrome differentiation and treatment of traditional Chinese medicine, acupoints are selected on the corresponding parts, which has a definite effect. It is regarded as a characteristic traditional Chinese medicine therapy for bronchial asthma. This protocol elaborates the methods and steps of patient management, material preparation, selection of acupoints, operation, and postoperative nursing to ensure safe and effective moxibustion treatment to significantly improve the clinical symptoms and quality of life of patients with bronchial asthma.

This protocol presents a curative procedure of moxibustion in treating patients with bronchial asthma.

Bronchial asthma is a chronic inflammatory disease of the airway which can lead to symptoms such as recurrent wheezing, shortness of breath, chest ...

Improving FSN Therapy in the Rat Model&#58; Minimizing Discomfort and Maximizing Efficiency

Efficacy of Fu's Subcutaneous Needling on Sciatic Nerve Pain: Behavioral and Electrophysiological Changes in a Chronic Constriction Injury Rat Model

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the mechanisms of action are still not fully understood. During FSN treatment, the FSN needle is inserted and retained in the subcutaneous tissues for a long duration with a swaying movement. However, challenges arise from maintaining a posture while manipulating FSN in animal models (e.g., rats) for researchers. Uncomfortable treatment can lead to fear and resistance to FSN needles, increasing the risk of injury and may even affect research data. Anesthesia may also affect the study results too. Hence, there is a need for strategies in FSN therapy on animals that minimize injury during the intervention. This study employs a chronic constriction injury model in Sprague-Dawley rats to induce neuropathic pain. This model replicates the pain induced by nerve injury observed in humans through surgical constriction of a peripheral nerve, mimicking the compression or entrapment seen in conditions such as nerve compression syndromes and peripheral neuropathies. We introduce an appropriate manipulation for easily inserting an FSN needle into the subcutaneous layer of the animal&#39;s body, including needle insertion and direction, needle retention, and swaying movement. Minimizing the rat&#39;s discomfort prevents the rat from being tense, which causes the muscle to contract and hinder the entry of the needle and improves the study efficiency.

Author Spotlight: Unveiling the Therapeutic Effects of FSN Treatment &#8211; Bridging Research and Clinical Applications in Neuropathic Pain

We present a protocol for using Fu's subcutaneous needling in a chronic constriction injury model to induce sciatic nerve pain in rats.

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the ...

Standardized Tuina Protocol for Rat DRG Compression Model

Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar disc herniation, spinal canal stenosis, and intervertebral foramen stenosis. Although Tuina, a traditional Chinese manual therapy, has shown analgesic effects in clinical practice for the treatment of neuropathic pain, its underlying neurobiological mechanisms remain unclear. Animal models are essential for elucidating the basic principles of Tuina. In this study, we propose a standardized Tuina protocol for rats with compression of the dorsal root ganglion (DRG), which involves inducing DRG compression by inserting a stainless steel rod into the intervertebral foramen, performing Tuina manipulation with specific parameters of location, intensity, and frequency in a controlled environment, and assessing the behavioral and histopathological outcomes of Tuina treatment. This article also discusses the potential clinical implications and limitations of the study and suggests directions for future research on Tuina.

Author Spotlight: Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain  

This article presents a manipulation for treating chronic compression of the dorsal root ganglion in rats using Tuina therapy, along with a method for evaluating its effectiveness based on pain behavior and histopathological results.

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar ...

Rat Jackets to Enhance Suspension Fixation During Electroacupuncture on Rats

Improved Suspension Fixation Using Rat Jacket in Rat Acupuncture Experiments

In basic experimental acupuncture research, rats are commonly used as laboratory animals. However, it is difficult for them to maintain a fixed posture. During electroacupuncture procedures, proper immobilization of rats is essential. Various methods of rat fixation are currently used, including anesthesia fixation, high-platform fixation, binding fixation, and fixation with a self-made rat coat. However, these methods have their limitations, which may affect the efficiency and operability of the experiment to some extent. This protocol introduces a method of suspending and fixing rats using rat clothes. Firstly, rats are clothed with rat jackets that match their body shape, taking advantage of their preference for darkness and burrowing. The needling operation can then be carried out after the rats have worn rat clothes. When suspended, the rats are relatively still, as their limbs cannot move. This fixation method offers not only economical and user-friendly benefits but also ensures a stable and reliable fixation of the rats in a comfortably relaxed position. It also effectively minimizes time consumption, experimental space, and manpower resources. Additionally, this method allows for the exposure of most acupoints used for acupuncture in rats. This article primarily concentrates on the device's composition, encompassing a specially designed rat jacket, an elevated fixation rack, and their connecting structures. Additionally, an illustrative example will be presented to demonstrate the application of the rat clothing-based suspension fixation method in rat electroacupuncture procedures.

Author Spotlight: Efficient Rat Clothing for Improved Rat Fixation During Electroacupuncture 

Here, we present an improved suspension fixation method using a rat jacket to effectively immobilize rats during acupuncture experiments. Furthermore, the structure and application of this fixation apparatus, as well as instructions and evaluation for its utilization in electroacupuncture therapy, is elucidated.

In basic experimental acupuncture research, rats are commonly used as laboratory animals. However, it is difficult for them to maintain a fixed posture. ...

Multicolor Immunofluorescence of the Nasal Tissue in an Allergic Rhinitis Rat Model

Immunofluorescent Labeling in Nasal Mucosa Tissue Sections of Allergic Rhinitis Rats via Multicolor Immunoassay

Allergic rhinitis (AR) is a chronic, non-infectious inflammatory disease of the nasal mucosa, primarily mediated by specific immunoglobulin E (IgE), affecting approximately 10%-20% of the world's population. While immunofluorescence (IF) staining has long been a standard technique for detecting disease-specific protein expression, conventional IF techniques are limited in their ability to detect the expression levels of three or more proteins in the same sample. Consequently, multicolor IF techniques have been developed in recent years, which allow the simultaneous labeling of multiple targets in cells or tissues.
This protocol provides a comprehensive overview of the process for establishing a rat model of AR, obtaining nasal mucosal samples, and the technical procedures for multicolor immunofluorescence. All rats in the AR group exhibited typical symptoms such as sneezing, a runny nose, and an itchy nose, with behavioral observations scoring &#8805;5 points. Hematoxylin and eosin (H&amp;E) staining revealed increased inflammatory cell counts and disrupted nasal mucosal integrity in the AR group. Multicolor immunofluorescence (mIF) demonstrated increased expression of ROR&#947;t and TICAM-1, while Foxp3 expression decreased in the nasal mucosa tissue of AR rats.

Author Spotlight: Advancing Allergic Rhinitis Research with Multicolor Immunofluorescence

This protocol describes a multicolor immunofluorescence technique for evaluating the rat model of allergic rhinitis.

Allergic rhinitis (AR) is a chronic, non-infectious inflammatory disease of the nasal mucosa, primarily mediated by specific immunoglobulin E (IgE), ...

Evaluating Acupoint Catgut Embedding Therapy for Allergic Rhinitis Management

Acupoint Catgut Embedding Therapy in Traditional Chinese Medicine for Managing Allergic Rhinitis

Acupuncture Point Embedding Therapy is a comprehensive treatment method that combines modern biomaterials with acupuncture techniques. Acupoint catgut embedding (ACE) not only avoids the adverse reactions associated with Western medicine but also incorporates the benefits of traditional acupuncture, extending the duration of its therapeutic effects. Allergic rhinitis (AR) is a condition known for its challenging treatment and tendency to persist. This randomized controlled trial included 128 AR patients to evaluate the effectiveness of ACE for AR management. Data were collected at baseline, four weeks after treatment, and eight weeks after treatment. Baseline analysis was performed, and the visual analog scale (VAS) was used as a clinical observation index for this method. The rhinoconjunctivitis quality of life questionnaire (RQLQ) served as a quality of life index. Specific IgE (sIgE), IL-4, IL-10, and IL-12 were used as laboratory observation indices. The results of the experiment validate the clinical effectiveness of ACE in treating AR. Although this technique is efficient and commonly used in clinics, its invasive nature poses risks of hospital-acquired infections. Based on this experiment, this protocol refines traditional techniques, providing detailed instructions on patient preparation, operation techniques, and postoperative care to ensure the safe and effective administration of therapy. By standardizing this therapy, ACE is anticipated to become an important non-drug treatment option for alleviating symptoms in AR patients, thereby significantly enhancing their quality of life.

This protocol details the procedure for acupoint burrowing and demonstrates its effectiveness in treating allergic rhinitis.

Acupuncture Point Embedding Therapy is a comprehensive treatment method that combines modern biomaterials with acupuncture techniques. Acupoint catgut ...

Effects of Different Connection Modes of Electroacupuncture on Electrocardiogram and Nerve Discharge in Rats

Electroacupuncture (EA) is one of the most commonly used methods in acupuncture and has a good effect on pain, depression, sensory movement disorders, and other diseases. The effectiveness of EA is influenced by many factors, such as the accuracy of acupoint selection, the duration and course of EA treatment, and EA parameters. However, it has rarely been discussed whether the positive and negative electrodes of the EA instrument with acupoints at different locations and distances have an effect on the curative effect. In this experiment, we observed the effects of connecting the EA instrument to acupoints at different positions and distances on ECG and vague nerve discharge in rats and preliminarily explained whether the electric field formed by different modes of EA connection has an effect on the function of the body. The connection modes of EA in this experiment included the same acupoints on both sides of the body, the same meridian or different meridian acupoints on the same side of the body, and two needles in the same acupoint area. The results showed that when the positive and negative poles were connected to the acupoints on both sides of the body, the recording of ECG and vagal nerve activity was disturbed (the same fore and hind limbs); when the acupoints were connected on the same side of the body, the smaller the distance between the two needles, the smaller the effect on ECG and vagal nerve activity recording, and the effect increased with increasing current; when the acupoints were in the same acupoint area, the recording of ECG and vagal nerve activity was not disturbed if the two needles did not form a short circuit.

Until now, there is no uniform method for electroacupuncture connection in clinical and fundamental research. We present a protocol that attempted to compare the effects of different connection methods on electrocardiogram (ECG) and neural discharge in rats to explore the most appropriate way to connect the electroacupuncture instrument.

Electroacupuncture (EA) is one of the most commonly used methods in acupuncture and has a good effect on pain, depression, sensory movement disorders, and ...

Acupuncture in a Rat Model of Asthma

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