This work was supported with funds from the National Natural Science Foundation of China (NSFC) Nos. 82374564 (to YJD) and Nos. 82074566 (to YJD) and from Sun Guojie Inheritance Base for TCM Acupuncture-Moxibustion of World Federation of Acupuncture-Moxibustion Societies in Wuhan, China (World Federation of Acupuncture-Moxibustion Societies [2019] No. 26) to YJD and the National Famous TCM Inheritance Studio construction project of the National Administration of Traditional Chinese Medicine, China (the National Administration of Traditional Chinese Medicine [2022] No. 5) to YJD.

All experiments were carried out following the National Institutes of Health - Guide for the Care and Use of Laboratory Animals and were approved by the Hubei University of TCM - Institutional Animal Care and Use Committee. Adult female Sprague-Dawley (SD) rats, weighing 220 &#177; 20 g, were used for the test. All rats were maintained on a 12 h light/dark cycle at 23 &#177; 1 &#176;C and 40%-50% humidity, with free access to food and water.
1. Pre-operative preparation
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Osteoporosis Management in Rats Through Lingnan Fire-Needle Therapy

The authors declare that they have nothing to disclose.

Lingnan fire-needle therapy has a profound historical background. Records on fire needles and needling methods can be traced back to Huangdi Neijing. Modern acupuncturists Situ Bell and Zhang Jiawei combined Lingnan&#39;s climate and the physical characteristics of its people to create Lingnan fire-needle therapy3. Lin Guohua summarized Lingnan Fire-Needle therapy, as warming, dredging, tonifying, clearing, and eliminating, with warming as the essence24.
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Chinese medicine believes that factors such as climate and environment can affect a person&#39;s physique, which in turn affects the occurrence and development of diseases. The Lingnan region of China, mainly including Guangdong, Guangxi, and Hainan Island, is located in the tropical monsoon climate region. As a result, people living in this region for a long time are prone to a form of spleen deficiency. In some cases, it spreads to the kidney and liver, leading to the prevalence of osteoporosis1. The spleen and stomach function in transporting and absorbing water and grains, which are the source of qi and blood biochemistry, while the liver helps maintain the marrows. So, bone nutrition relies on the supply of the spleen, stomach, and liver. In Chinese medicine, the kidney is capable of generating marrow and strengthening bones, and its vitality directly affects the strength and function of bones. Therefore, kidney deficiency is directly related to osteoporosis. According to TCM, liver (Gan)-kidney (Shen) deficiency and lack of essence (Jing) are the underlying causes. In some cases, malnutrition may occur due to a spleen (Pi) and stomach (Wei) deficiency or an imbalance of the two. That is, the deficiency of the liver (Gan), kidneys (Shen), spleen (Pi), and stomach (Wei) are the root causes, while external qi (Xie qi) blocks meridians and damages bones and muscles. According to the World Health Organization (WHO), OP is a medical condition characterized by low bone mass and tissue deterioration2, which is a serious concern for the elderly. The selection of appropriate interventions is crucial for the prevention and treatment of osteoporosis.
Modern acupuncturists have developed a unique Lingnan characteristic of fire-needling therapy by combining the climate of Lingnan and the physical characteristics of Lingnan people3. Fire needle therapy is a kind of treatment method in which special needles are burned red and quickly pierced into specific parts or acupoints of the human body to treat various ailments. This method is widely used in skin diseases, musculoskeletal and joint diseases, neurological diseases, and other systemic diseases and shows good efficacy4. The treatment of osteoporosis with fire needles has been used in clinics for a long time, and it has a better curative effect than simple acupuncture or oral administration of traditional Chinese medicine5,6,7. The Lingnan fire-needle therapy integrates acupuncture, three-edged needles, and scarring moxibustion. It can improve the qi and blood circulation in the meridians while warming the kidney. This treatment can address both blood stasis and kidney (Shen) deficiency, which contribute to the pathogenesis of OP in TCM. What&#39;s more, it has the advantages of being low cost, having quick effects, a short treatment period, and being safe. However, there is a lack of research on its mechanism. This paper introduces a specific procedure of Lingnan Fire-Needle in osteoporosis model rats and provides a methodological basis for conducting related basic research.

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	<tbody>
		<tr>
			<td>75% Alcohol</td>
			<td>Shandong Lircon medicaltechnology co., ltd</td>
			<td></td>
			<td>Disinfection</td>
		</tr>
		<tr>
			<td>95% Alcohol</td>
			<td>Shandong Lircon medicaltechnology co., ltd</td>
			<td></td>
			<td>Burn needles</td>
		</tr>
		<tr>
			<td>Adult female Sprague-Dawley (SD) rats</td>
			<td>Liaoning Changsheng biotechnology co., Ltd.</td>
			<td>SCXK 2020-0001</td>
			<td>Modeling of osteoporosis</td>
		</tr>
		<tr>
			<td>Deluxe thermostat heat mat</td>
			<td>Zhenhua Instruments</td>
			<td></td>
			<td>For maintaining the body temperature of rats.</td>
		</tr>
		<tr>
			<td>Diclofenac sodium injection</td>
			<td>Shandong Province Jianmu Bio-pharmaceutical Co.</td>
			<td>150182913</td>
			<td>Antipyretic and analgesic</td>
		</tr>
		<tr>
			<td>Elevated fixation rack</td>
			<td>Taizhou Shi Ai Scientific and Educational Instruments and Equipment Co.</td>
			<td></td>
			<td>Fix rats</td>
		</tr>
		<tr>
			<td>Fire needles</td>
			<td>Beijing Luoya Shanchuan Medical Equipment Co., Ltd.</td>
			<td>20190240</td>
			<td>A tool of needling</td>
		</tr>
		<tr>
			<td>Hemostatic forceps</td>
			<td>Qingdao Schultz Biotechnology Co.</td>
			<td></td>
			<td>Fix rats</td>
		</tr>
		<tr>
			<td>Liguter</td>
			<td>Shanghai Machi Trading Co.</td>
			<td>H5</td>
			<td>Ignite alcohol</td>
		</tr>
		<tr>
			<td>Lodophor disinfecting solution</td>
			<td>Qingdao Hai&#39;s Hainuo Inovation Disinfection Technology Co., Ltd.</td>
			<td>2016-0604</td>
			<td>Disinfection</td>
		</tr>
		<tr>
			<td>Medical cotton ball</td>
			<td>Jiangsu Misawa Medical Supplies Co., Ltd.</td>
			<td>20172142357</td>
			<td>Disinfection</td>
		</tr>
		<tr>
			<td>pentobarbital sodium</td>
			<td>Beijing Jiehui Biotechnology Co.</td>
			<td>P3761</td>
			<td>anesthesia</td>
		</tr>
		<tr>
			<td>Pet electric clipper</td>
			<td>Fenghua Xikou Lubao Knives &#38; Scissors Electrical Appliance Factory</td>
			<td></td>
			<td>Shave the hair of rats</td>
		</tr>
		<tr>
			<td>Self-made rat coat</td>
			<td></td>
			<td></td>
			<td>Fix rats</td>
		</tr>
		<tr>
			<td>Tweezers</td>
			<td>Qingdao Schultz Biotechnology Co.</td>
			<td></td>
			<td>Pinch cotton balls</td>
		</tr>
	</tbody>
</table>

operation procedure and precautions of lingnan fire-needle therapy in osteoporosis model rats

Compared to filiform needle therapy, fire-needle therapy has both the stimulation of needles and the warming effect of heat, making it have unexpected effects on some chronic diseases and incurable diseases. Osteoporosis (OP) has a high incidence in postmenopausal women and middle-aged and elderly men, and the treatment cycle is long. According to Traditional Chinese Medicine (TCM), Lingnan fire-needle therapy has shown potential in treating osteoporosis. However, there is still a long way to go before it can be widely used. This article focuses on the application of Lingnan fire-needle therapy in the intervention of OP in rats. It covers the selection of needle tools, acupuncture point selection, positioning of rats' bodies, and fixation methods. We also outline the steps and precautions to be taken during and after needling with fire needles. The experiment was done with three groups: a normal group, a model group, and a fire-needle group, each containing 10 rats. The rats in the fire-needle group were treated with fire-needle intervention for six sessions. After the intervention period, we collected femoral specimens and performed micro-CT scans. The results suggest that fire needling can enhance bone morphology and mineral density in OP rats. This information can serve as a methodological basis for conducting basic research on fire-needle therapy.

Author Spotlight: Exploring the Impact of Lingnan Fire-Needle Therapy in Osteoporosis Intervention

Operation Procedure and Precautions of Lingnan Fire-Needle Therapy in Osteoporosis Model Rats

Our research is dedicated to exploring the therapeutic effects and the mechanisms of acupuncture in the management of age-related and chronic conditions. Specifically, I'm investigating the impact of Lingnan fire-needle therapy on osteoporosis in golden models. The existing literature on osteoporosis treatments is predominantly focused on pharmacological interventions.Our protocol aims to fill the gap by examining a non-pharmacological approach, Lingnan fire-needle therapy, for its potential in improving bone health. Compared to conventional treatments, the Lingnan fire-needle therapy offers a non-invasive, drug-free alternative with minimal side effects. It integrates traditional acupuncture with the unique application of heat, potentially enhancing therapeutic outcomes for osteoporosis.Our findings could provide empirical evidence supporting the use of fire-needle acupuncture in osteoporosis treatment, thereby enriching the body of research on integrative medicine, and offering new insights into the integration of traditional practices in modern healthcare. In the future, our laboratory will concentrate on elucidating the molecular and cellular pathways through which fire-needle acupuncture exerts its effect. We aim to extend our research to include a broader range of geriatric and chronic diseases, further validating the therapies efficacy and applicability.

The data in Table 1 shows that compared with the normal group, the thickness of trabecular bone (Tb.Th), the number of trabecular bone (Tb.N) and the static parameter bone volume fraction of trabecular bone (BV/TV) in the model group were all significantly reduced (p &#60;0.05), and the separation of trabecular bone (Tb.Sp) was elevated (p &#60;0.05); indicating that the bone structure of trabecular bone in trabecular bone-enriched areas of the femoral epiphysis of the rat was significantly disrupted aft...

Read this Scientific Article Protocol about Author Spotlight: Exploring the Impact of Lingnan Fire-Needle Therapy in Osteoporosis Intervention at JoVE.com

Here, we present a protocol for the application of Lingnan Fire-Needle therapy in the intervention of osteoporosis model rats.

Compared to filiform needle therapy, fire-needle therapy has both the stimulation of needles and the warming effect of heat, making it have unexpected ...

operation-procedure-precautions-lingnan-fire-needle-therapy

Research

JoVE Journal

Medicine

Fire-Needle Therapy to Treat Osteoporosis in Rat Models

To begin, position the oophorectomized menopause model rat on a non slippery mat and locate the acupoints for the treatment. Using a pet electric clipper, shave the acupuncture sites and mark them with an oil-based marker. With the left hand, hold the rat's tail and gently pull back.Open the port of the rat coat with the right hand, and carefully let the rat burrow into it. When the rat reaches the middle of the coat, hold both sides of the rat's shoulder blades with the right hand and pull out the rat's forelimbs with the left hand. After tightening the ties on the front of the rat coat, position the hind limbs of the rat outside the coat.For the swing type suspension, fix two S hooks into the knot on the back of the rat coat and hang them on the iron frame to suspend the rat. Sterilize the local area of the rat three times with 5%ionophore, followed by 75%alcohol. Use hemostatic forceps to hold a cotton ball soaked in 95%alcohol and light it with a lighter.Then, with the right hand, hold the acupuncture needle between the thumb, forefinger, and middle finger. Hold both the needle tip and the body of the needle onto the flaming cotton ball until it turns red. Next, holding the needle vertically above the acupoint, quickly puncture the point and pull out within one second.Finally, press a sterile dry cotton ball to the needle hole to prevent bleeding. Compared with the normal group, a large number of trabeculae were lost in the femur cancellous matrix in the model group. However, there was a large increase in the thickness of trabeculae in the center and periphery of the cancellous bone of the femur in the fire needle group, as compared to the model group.

317 Views.  Hubei University of Chinese Medicine.  Here, we present a protocol for the application of Lingnan Fire-Needle therapy in the intervention of osteoporosis model rats.

Osteoporosis Management in Rats Through Lingnan Fire-Needle Therapy

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