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 Provincial Center for Disease Control and Prevention Laboratory Animal Management and Use Committee. A total of 18 3-month-old female Sprague-Dawley (SD) rats weighing 220-250 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. Following a one-week acclimatization period, during which the rats were allowed to habituate to the laboratory conditions, the experiments were initiated.
1. Model development
<ol>
	<li>The experimental design is shown in Figure 1. Randomly divide 18 Sprague-Dawley 3-month-old female rats weighing 220-250 g into control, model, and TEAT groups. Each group consists of 6 rats.</li>
	<li>Inject the model group and TEAT group daily with D-galactose (D-gal; 120 mg/kg/day), while the control group receives saline injections (120 mg/kg/day), for 7 weeks (49 days) in the lower ventral area22. The injection should be administered daily between 8:00 and 9:00 a.m.</li>
	<li>Weigh the rats on the 7th day of each week between 9:00 and 10:00 a.m. The experiment lasted for 49 days. Bury the threads every 14 days on the 14th, 28th, and 42nd days. Fix the time for each thread embedding between 9:00-12:00 am.</li>
	<li>Continue intraperitoneal administration of D-galactose for 7-weeks to induce Alzheimer&#39;s disease-related pathological manifestations in rats22.</li>
</ol>
2. Preparation for TEAT
<ol>
	<li>Sterilize surgical instruments such as ophthalmic scissors, trays, and tweezers and disinfect. Clean the surfaces in the operating room environment thoroughly and disinfect.</li>
	<li>Prepare the surgical platform by laying a small animal heating pad flat on the surface, followed by draping with a sterile towel. Subsequently, place the cleaned and disinfected operating board smoothly on top, with the rough side facing upwards. Then, switch on the heating pad and set the temperature to 32 &#176;C.</li>
	<li>For the procedure, select a No. 6 disposable thread-embedding needle with a diameter of 0.6 mm and a 5-0 absorbable surgical suture (collagen suture) with a length of 2 cm. Cut the 2 cm 5-0 absorbable surgical suture into 0.5 cm and 0.2 cm pieces using ophthalmic scissors. Soak the cut thread in 0.9% saline and set aside.</li>
	<li>Wash hands with soap and water for 20 s, and then dry them. Afterward, wear sterile gloves.</li>
	<li>Anesthetize the rats with an IP injection of chloral hydrate (300 mg/kg) in the lower ventral area, ensuring that anesthesia is confirmed when animals are unresponsive in a surgical plane, based on the lack of response from a toe pinch test, and maintaining a stable physiological state. Additionally, to prevent corneal dryness and potential damage while the rats are under anesthesia, apply a generous amount of veterinary ointment to the eyes, ensuring they remain lubricated throughout the procedure. 
	NOTE: It is recommended to anesthetize the rat, as it is painful for them to experience the piercing.</li>
</ol>
3. Thread-embedding operation for Baihui (GV 20)
<ol>
	<li>Locate the acupoint Baihui (GV 20) in rats at the right midpoint of the parietal bone (Figure 2A). Position the rats to fully expose the head, with the ears pulled forward to approximate the method used in humans for connecting the ear apex lines. Identify the Baihui (GV 20) point by palpating the central longitudinal ridge of the skull and pinpointing the location roughly at its midpoint. Confirm this acupoint further at the level where the ears would make contact with the head if laid flat, aligning with the intersection of the sagittal suture and the parietal bone. Use a marker to mark Baihui (GV 20).</li>
	<li>Shave the head area around the acupoint of the rat using a trimmer (usually an electric men&#39;s beard trimmer).</li>
	<li>Prior to surgery, disinfect the acupoints and surrounding skin by applying a 0.5% iodine solution in circular motions, starting from the center, and moving outwards. Subsequently, perform a triple disinfection process: use the iodophor 3x, each followed by an application of alcohol to ensure proper decontamination and prepare the site for aseptic technique.</li>
	<li>Use forceps to clamp out the 0.2 cm absorbable surgical suture soaked in saline and place it in the front end of the needle tube. Then, connect it to the needle core.</li>
	<li>Pinch the area around Baihui (GV 20) with thumb and index finger on one hand, while holding the needle with the other.</li>
	<li>Insert the needle horizontally and quickly at a 15&#176; angle to the skin surface, 0.1 cm below the Baihui (GV 20) point, while pinching the skin (Figure 3A).</li>
	<li>After the needle tip reaches 0.1 cm above the Baihui (GV 20) point, push the needle core while retracting the needle tube to fully implant the 0.2 cm absorbable surgical suture into the subcutaneous tissue of the acupoint (Figure 3B).</li>
	<li>Slowly remove the needle, making sure no suture is exposed. Press the needle hole with sterile dry cotton for 10 s to prevent bleeding. If the thread is outside the body after embedding the line at acupoints, extract it and re-operate.</li>
</ol>
4. Thread-embedding operation for Shenshu (BL 23)
<ol>
	<li>The acupoint Shenshu (BL 23) in rats is located 5 mm lateral to the 2nd lumbar vertebra (Figure 2B). First, locate the 6th lumbar vertebra based on the hip tubercle of the rat. Then locate the 2nd lumbar vertebra by counting upward four vertebral bodies. Shenshu (BL 23) will be 5 mm lateral to it23. 
	NOTE: The combination of the Baihui (GV 20) and Shenshu (BL 23) acupoints is a frequently utilized set in Traditional Chinese Medicine (TCM) theory for the prevention and treatment of cerebral disorders. As confirmed by previous research20,21, this acupoint pairing has been established for its efficacy.</li>
	<li>Use a marker to mark Shenshu (BL 23). Shave the back and waist area around the acupoint of the rat using a trimmer (usually an electric beard trimmer).</li>
	<li>Prior to surgery, disinfect the acupoints and surrounding skin by applying 0.5% iodine solution in a circular motion, starting from the center and moving outwards. Subsequently, perform a triple disinfection process: use the iodophor three times, each followed by an application of alcohol to ensure proper decontamination and preparation of the site for aseptic technique.</li>
	<li>Use forceps to clamp out the 0.5 cm absorbable surgical suture soaked in saline and place it in the front end of the needle tube. Then, connect it to the needle core.</li>
	<li>Stretch the skin with the index and middle fingers of the pressing hand or the thumb and index fingers. Hold the needle with the puncturing hand.</li>
	<li>Quickly insert the needle perpendicularly into the point 0.25 cm below the Shenshu (BL 23). See Figure 4A.</li>
	<li>After the needle tip reaches 0.5 cm under the skin, adjust the needle tip, making the direction of insertion centripetal, forming a 45&#176; angle with the skin surface and obliquely insert 0.5 cm (Figure 4B). 
	NOTE: It is important to exercise caution when needling the Shenshu (BL 23), which is located near the kidney. Deep insertion or repeated lifting and thrusting of the needle can cause damage. Therefore, we recommend an insertion depth of around 0.3-0.6 cm.</li>
	<li>Push the needle core while retracting the tube and fully implant the 0.5 cm absorbable surgical suture into the acupoint&#39;s muscle tissue of the acupoint (Figure 4C).</li>
	<li>Slowly pull out the needle, ensuring no suture is exposed. Then, press the needle hole with sterile dry cotton for 10 s to prevent bleeding. If the thread is outside the body after embedding the line at acupoints, extract it and re-operate.</li>
</ol>
5. Post-operative care
<ol>
	<li>Postoperatively, place the rat on a small animal heating pad set at a temperature of 32&#176;C to facilitate recovery from anesthesia.Ensure the animal is fully conscious and its condition is satisfactory before returning it to its housing enclosure.</li>
	<li>After surgery, observe signs of sterile inflammatory responses in the affected area within a week. These signs may include redness, swelling, or a pus-like discharge. Typically, these symptoms do not require any treatment. However, if there is an excessive amount of exudate, drain and disinfect the affected area. In such cases, apply erythromycin ointment until the wound fully recovers.</li>
</ol>
6. Behavioral testing and histological examination
<ol>
	<li>Following the intervention, at week 8, perform the Morris Water Maze (MWM) experiment2,24 to evaluate their learning and memory capabilities. Perform the learning trials over 5 days, with four trials per day. Then administer a probe trial after 24 h from the last acquisition day.</li>
	<li>Perform statistical comparisons using statistical analysis software. Analyze the behavioral data of learning ability by one-way analysis of variance (ANOVA) and post-hoc Tukey&#39;s test. Assess the significance of escape latency data by two-way repeated measure analysis of variance (ANOVA) followed by the post-hoc Bonferroni test. A value of p &#60;0.05 is considered significant in all tests.</li>
	<li>After the behavioral assessment, anesthetize the rats as above and euthanize with intraperitoneal injections of 150 mg/kg body weight of pentobarbital sodium.</li>
	<li>Fix the rat&#39;s brain through perfusion using 4% paraformaldehyde perfused via the vascular system25.
	<ol>
		<li>Insertion and fixation of the gavage needle
		<ol>
			<li>Prepare a 12G needle and ensure it is straight and suitable for passing through the ventricle of the rat. Locate the ventricle of the rat, which is typically the left ventricle, and insert the needle through it into the ascending aorta. This step requires precision and a good understanding of rat anatomy.</li>
			<li>Once the needle is in place, use a hemostat to clamp the gavage needle near the point where it enters the aorta. This will help to secure the needle and prevent it from moving.</li>
			<li>Ensure that the clamp is tight enough to hold the needle securely but not so tight as to damage the aorta or impede blood flow. The gavage needle should remain in the ascending aorta throughout the procedure to maintain proper perfusion.</li>
		</ol>
		</li>
		<li>Perfusion with normal saline
		<ol>
			<li>Prepare a 50cc syringe without a needle for the perfusion. Attach the other end of the gavage needle, which is outside the rat&#39;s body, to the syringe filled with normal saline.</li>
			<li>Make a small incision to cut the right atrial appendage to allow blood to flow out of the heart. This step ensures that the blood is displaced by the saline during perfusion.</li>
			<li>Begin injecting the normal saline into the ascending aorta at a steady and constant rate. This will flush out the blood and prepare the tissues for fixation.</li>
			<li>Monitor the fluid coming out of the right atrial appendage and be prepared to change the syringe as needed to maintain a continuous perfusion with fresh saline. 
			&#8203;placed, prepare to perfuse with the fixative.</li>
			<li>Replace the syringe containing saline with one containing 4% paraformaldehyde, a commonly used fixative in histology. Proceed with the perfusion using the paraformaldehyde to fix the tissues in place for subsequent processing and analysis. 
			NOTE: Throughout this procedure, it is crucial to maintain aseptic techniques to prevent infection and to handle the rat gently to minimize injury. Additionally, all personnel involved should be trained in the technique to ensure consistency and effectiveness.</li>
		</ol>
		</li>
	</ol>
	</li>
	<li>Remove the rat brain and place it in a specimen bottle containing 4% paraformaldehyde for 24-48 h at 4 &#176;C25,26. Perform ethanol dehydration and xylene treatment step by step, followed by conventional continuous coronal sections (4-5 &#181;m) for measurement. Evaluate the morphology and loss of hippocampal neurons in each group using Hematoxylin and Eosin (H&#38;E) staining (200x). 
	NOTE: A necropsy examination to assess for gastrointestinal (GI) lesions was not conducted on the rodents based on the absence of clinical signs indicative of GI disturbances throughout the experimental period. The animals exhibited no overt adverse reactions that would suggest the presence of such pathologies.</li>
</ol>

Improving Cognitive Functions in Rats via Thread-Embedding Acupuncture Therapy

The authors declare that they have nothing to disclose.

Intervention time 
The etiology of AD is complex, and its pathogenesis remains uncertain. Currently, there is no effective treatment for the disease, and its course is irreversible. Therefore, the academic community has reached a consensus on the importance of focusing on the prevention and treatment of AD. The prevention and treatment strategies for AD emphasize early intervention, following the TCM concept of treating conditions before they occur, emphasizing treatment before its onset, transmissi...

Thread-embedding acupuncture (TEA) involves embedding catgut or absorbable sutures into acupoints using a special needle. This technique has a prolonged effect until the sutures are absorbed and degraded. It provides continuous and sustained stimulation to the acupoints for a week or longer1, achieving a comparable outcome without the necessity for daily repetitive procedures2. This reduces the number of patients seeking medical treatment, thus conserving medical resources to some extent. In recent years, TEAT has been increasingly used in various medical fields in China, including internal medicine, external medicine, dermatology, facial features, gynecology, and pediatrics3. Studies have shown that it is effective in producing long-term and reliable effects for certain chronic and complex diseases, such as epilepsy4 and facial nerve paralysis5. It has the advantages of minimal trauma and simple operation6. Additionally, the treatment costs of TEAT have been reported to be lower than those of electroacupuncture (EA)7. Therefore, TEAT is increasingly valued in clinical and scientific research.
Alzheimer's disease (AD) is a neurodegenerative condition that progresses over time. Its pathophysiological process begins to accumulate approximately 20 years before clinical symptoms appear, but its mechanism is still not fully understood8. Given the limited availability of effective drugs for AD, research is increasingly focusing on the prodromal and preclinical stages of the disease9. Prevention is a crucial strategy for addressing the development and progression of AD9,10. In recent years, traditional Chinese medicine (TCM) has made significant progress, and it is expected to offer new possibilities for preventing and treating AD11,12. As a subtype of acupuncture, TEAT is preferred over other therapies due to its wide treatment range, minimal trauma, simple operation, low treatment frequency, and long-lasting efficacy6. Emerging literature suggests that TEAT may benefit individuals with obesity13,14, diabetes15, insomnia16, postmenopausal osteoporosis17, and depression18, all of which are risk factors for AD. It also improves senescence in aging rats by regulating mitophagy2, improves spatial learning and memory impairment, alleviates pathological damage of the hippocampus, and inhibits inflammation response in vascular dementia (VD) rats19. It appeared attractive to figure out whether TEAT could also act on pathological processes during AD.
The current research lacks uniformity and standardization in several critical elements of TEAT operation, including the embedding method, embedding tool, thread, and embedding interval period. This lack of standardization affects the evaluation of the therapy's effect and the exploration of related mechanisms and restricts external promotion and communication. Prior research20,21 has demonstrated that intraperitoneal administration of D-galactose (D-gal) can induce Alzheimer's disease-like pathological changes in rats, including neuronal degeneration and cognitive impairments. Furthermore, electroacupuncture at the Baihui(GV20) and Shenshu(BL23) has been shown to effectively ameliorate cognitive decline, mitigate neuroinflammation, and reduce neuronal damage. This study takes Alzheimer's disease-like model rats as an example to introduce the selection of acupoints and the operation process and precautions of TEAT under this model. It discusses the critical operational elements of TEAT in experimental research to provide a reference for future studies.

<table><tbody><tr><td>0.9% saline, 4% paraformaldehyde, Hematoxylin, Eosin Y (water-soluble), absolute ethanol, 0.01M PBS powder, xylene, hydrochloric acid, embedded paraffin, neutral gum</td><td>sinopharm (www.sinopharm.com)</td><td></td><td></td></tr><tr><td>5-0 absorbable surgical suture</td><td>bodamedical (www.bodamedical.com)</td><td></td><td></td></tr><tr><td>70% alcohol, iodophor, cotton ball</td><td></td><td></td><td>Used for disinfection.</td></tr><tr><td>chloral hydrate</td><td>sigma</td><td>C8383</td><td></td></tr><tr><td>coverslips, glass slides</td><td>Guangzhou Wuqiang Experimental Equipment Co., Ltd</td><td></td><td></td></tr><tr><td>Dehydrator</td><td>Wuhan Junjie JT-12J Computerized Biological Tissue Dehydrator</td><td></td><td></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>D-galactose</td><td>sigma</td><td>G0750</td><td></td></tr><tr><td>electric men&#039;s beard trimmer</td><td></td><td></td><td>Used for shaving rats.</td></tr><tr><td>Electronic scale for small animals (ZK-DST)</td><td>Henan Zhike Information Technology Co., Ltd</td><td></td><td></td></tr><tr><td>erythromycin ointment</td><td>frontpharm</td><td></td><td></td></tr><tr><td>glass dish</td><td></td><td></td><td>Used to place cut absorbable sutures.</td></tr><tr><td>High-Pressure Steam Sterilization Pot (YM100Z)</td><td>Shanghai Sanshen medical equipment factory</td><td></td><td>For sterilization of surgical instruments.</td></tr><tr><td>Ice Maker (CMB40)</td><td>Shanghai Sheyan Instrument Co., Ltd.</td><td></td><td></td></tr><tr><td>Microscope (BX-53)</td><td>OLYMPUS</td><td></td><td></td></tr><tr><td>milk powder</td><td></td><td></td><td>Used for the MWM test.</td></tr><tr><td>Morris Water Maze Video Tracking System &amp;amp; Data Analysis Software (Version: Watermaze 2.0)</td><td>Chengdu Taimeng Technology Co., Ltd</td><td></td><td></td></tr><tr><td>No.6 thread-embedding needle</td><td>bodamedical (www.bodamedical.com)</td><td></td><td></td></tr><tr><td>ophthalmic scissors, trays, tweezers</td><td></td><td></td><td></td></tr><tr><td>Pathology Microtome&amp;nbsp; (RM 2016)</td><td>LEICA, Germany</td><td></td><td></td></tr><tr><td>pentobarbital</td><td>Germany</td><td>P11011</td><td></td></tr><tr><td>permanent marker</td><td></td><td></td><td>Used to mark the acupoint.</td></tr><tr><td>Slicing knives (R35)</td><td>Feathers, Japanese</td><td></td><td></td></tr><tr><td>soap</td><td></td><td></td><td>Used to wash hands.</td></tr><tr><td>sterile gloves</td><td></td><td></td><td></td></tr></tbody></table>

operation procedure and precautions of thread-embedding acupuncture therapy in alzheimer

Thread-embedding therapy (TEAT) is a treatment that prevents and manages diseases by inserting a biodegradable suture into an acupoint, providing long-lasting stimulation. TEAT is a simple approach that avoids the discomfort of regular acupuncture and provides sustained therapeutic effects. This article discusses the potential impact of TEAT on the learning and memory abilities of rats with Alzheimer&#39;s disease-like symptoms. Since chemically induced neuronal degeneration and cognitive impairments in rats does not entirely reflect the true pathological changes observed in Alzheimer&#39;s disease. Consequently, our research group has designated these manifestations as Alzheimer&#39;s disease-like symptoms. A protocol has been established to outline the selection of acupoints, the operation process, and necessary precautions for the head and lower back. The experiment was conducted on three groups: a control group, a model group, and a TEAT group, each containing 6 rats. To induce Alzheimer&#39;s disease-like symptoms, rats were intraperitoneally injected with D-galactose for 7 weeks (49 days). The rats in the TEAT group received acupoint catgut embedding treatment. Following the intervention period, a Morris Water Maze (MWM) was conducted to evaluate the rats&#39; learning and memory. Subsequently, the rats were sacrificed, and their brain tissue was examined. A histological examination was performed to understand the effects of TEAT on the pathology of rats exhibiting symptoms of Alzheimer&#39;s disease. This study suggests that TEAT may improve learning and memory in rats with Alzheimer&#39;s disease-like symptoms, indicating a potentially promising new treatment approach for this neurodegenerative condition.

This article introduces the specific operation method of TEAT in Alzheimer&#39;s disease-like rats. It discusses the critical operational elements of TEAT in experimental research to provide a reference for future studies.
TEAT improves learning in Alzheimer&#39;s disease-like rats 
As illustrated in the flowchart (Figure 5), during the adaptive training period, it was observed that rats in each group were observing the surrounding environmen...

Read this Scientific Article Protocol about Author Spotlight: Exploring Acupuncture in Alzheimer's Research from Thread-Embedding Techniques to Clinical Trials at JoVE.com

Author Spotlight: Exploring Acupuncture in Alzheimer's Research from Thread-Embedding Techniques to Clinical Trials

This study presents a protocol for thread-embedding acupuncture therapy in Alzheimer's disease-like rats.

Operation Procedure and Precautions of Thread-Embedding Acupuncture Therapy in Alzheimer

Thread-embedding therapy (TEAT) is a treatment that prevents and manages diseases by inserting a biodegradable suture into an acupoint, providing ...

operation-procedure-precautions-thread-embedding-acupuncture-therapy

Research

JoVE Journal

Neuroscience

1.4K Views.  Hubei University of Chinese Medicine. This study presents a protocol for thread-embedding acupuncture therapy in Alzheimer's disease-like rats.

Video: Author Spotlight: Exploring Acupuncture in Alzheimer's Research from Thread-Embedding Techniques to Clinical Trials

Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting

Two-dimensional gel electrophoresis (2DE) is a powerful tool to uncover proteome modifications potentially related to different physiological or pathological conditions. Basically, this technique is based on the separation of proteins according to their isoelectric point in a first step, and secondly according to their molecular weights by SDS polyacrylamide gel electrophoresis (SDS-PAGE). In this report an optimized sample preparation protocol for little amount of human post-mortem and mouse brain tissue is described. This method enables to perform both two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and mini 2DE immunoblotting. The combination of these approaches allows one to not only find new proteins and/or protein modifications in their expression thanks to its compatibility with mass spectrometry detection, but also a new insight into markers validation. Thus, mini-2DE coupled to western blotting permits to identify and validate post-translational modifications, proteins catabolism and provides a qualitative comparison among different conditions and/or treatments. Herein, we provide a method to study components of protein aggregates found in AD and Lewy body dementia such as the amyloid-beta peptide and the alpha-synuclein. Our method can thus be adapted for the analysis of the proteome and insoluble proteins extract from human brain tissue and mice models too. In parallel, it may provide useful information for the study of molecular and cellular pathways involved in neurodegenerative diseases as well as potential novel biomarkers and therapeutic targets.

A common protein extraction protocol using urea/thiourea/SDS buffer for human and mice brain tissue allows indentification of proteins by 2D-DIGE and their subsequent characterization by mini 2DE immunoblotting. This method enables one to obtain more reproducible and reliable results from human biopsies and experimental models.

Two-dimensional gel electrophoresis (2DE) is a powerful tool to uncover proteome modifications potentially related to different physiological or ...

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

Assessment of Dendritic Arborization in the Dentate Gyrus of the Hippocampal Region in Mice

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity and extent of dendritic arborization correlates well with the synaptic plasticity in these cells. A reliable method for assessment of dendritic arborization is needed to characterize the deleterious effects of neurological disorders on these structures and to determine the effects of therapeutic interventions. However, quantification of these structures has proven to be a formidable task given their complex and dynamic nature. Fortunately, sophisticated imaging techniques can be paired with conventional staining methods to assess the state of dendritic arborization, providing a more reliable and expeditious means of assessment. Below is an example of how these imaging techniques were paired with staining methods to characterize the dendritic arborization in wild type mice. These complementary imaging methods can be used to qualitatively and quantitatively assess dendritic arborization that span a rather wide area within the hippocampal region.

We describe two methods for visualization and quantification of dendritic arborization in the hippocampus of mouse models: real-time and extended depth of field imaging. While the former method allows sophisticated topographical tracing and quantification of the extent of branching, the latter allows speedy visualization of the dendritic tree.

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity ...

Intracerebroventricular Injection of Amyloid-&#946; Peptides in Normal Mice to Acutely Induce Alzheimer-like Cognitive Deficits

Amyloid-&#946; (A&#946;) is a major pathological mediator of both familial and sporadic Alzheimer's disease (AD). In the brains of AD patients, progressive accumulation of A&#946; oligomers and plaques is observed. Such A&#946; abnormalities are believed to block long-term potentiation, impair synaptic function, and induce cognitive deficits. Clinical and experimental evidences have revealed that the acute increase of A&#946; levels in the brain allows development of Alzheimer-like phenotypes. Hence, a detailed protocol describing how to acutely generate an AD mouse model via the intracerebroventricular (ICV) injection of A&#946; is necessary in many cases. In this protocol, the steps of the experiment with an A&#946;-injected mouse are included, from the preparation of peptides to the testing of behavioral abnormalities. The process of preparing the tools and animal subjects before the injection, of injecting the A&#946; into the mouse brain via ICV injection, and of assessing the degree of cognitive impairment are easily explained throughout the protocol, with an emphasis on tips for effective ICV injection of A&#946;. By mimicking certain aspects of AD with a designated injection of A&#946;, researchers can bypass the aging process and focus on the downstream pathology of A&#946; abnormalities.

The amyloid-&#946; (A&#946;)-injected animal model enables the administration of a defined quantity and species of A&#946; fragments and reduces individual differences within each study group. This protocol describes the intracerebroventricular (ICV) injection of A&#946; without stereotactic instruments, enabling the production of Alzheimer-like behavioral abnormalities in normal mice.

Amyloid-&#946; (A&#946;) is a major pathological mediator of both familial and sporadic Alzheimer's disease (AD). In the brains of AD patients, ...

Acupoint Needle-Embedding Combined with Ironing Therapy for Postoperative Pain After Anal Surgery

Acupoint needle-embedding combined with ironing therapy is a non-drug treatment method to release postoperative pain after anal surgery. The practice is guided by traditional Chinese medicine (TCM) syndrome differentiation theory and employs acupoint stimulation and heat to alleviate pain. Although prior research has shown that these are dependable methods for pain relief, the combined effect of the two techniques has not been described. In our research, we found that compared to using diclofenac sodium enteric-coated capsules alone, adding acupoint needle-embedding combined with ironing therapy was more effective for reducing pain levels at different stages after hemorrhoid surgery. Although this technique is efficient and commonly used in clinics, due to its invasive practice, acupoint needle embedding still carries risks relating to hospital-acquired infections and broken needles. Ironing therapy, on the other hand, can result in burns and connective tissue injuries. Therefore, there is an urgent need to develop a standardized protocol for medical staff. Our protocol refines the traditional techniques and provides detailed instructions on patient preparation, operation techniques, and postoperative care to ensure the therapy is carried out safely and efficiently. By standardizing this therapy, this technique is expected to become an important complementary therapy for postoperative pain relief&#160;in hemorrhoids, which will significantly improve patients &#39;life quality after anal surgery.

The high incidence of pain after anal surgery is due to the physiological structure and pathological status of the anus. At present, the use of painkillers cannot provide satisfactory pain relief, which makes complementary therapy important. This paper proposes acupoint needle-embedding combined with ironing therapy to relieve postoperative pain.

Acupoint needle-embedding combined with ironing therapy is a non-drug treatment method to release postoperative pain after anal surgery. The practice is ...

Medicine

Scalp Acupuncture Synchronizing Dual Task Gait for Enhancing Prefrontal Cortex Response

Studies have shown that motor-cognitive dual task can greatly improve motor/cognitive function. However, the therapeutic effect of motor-cognitive dual task is still limited. How to improve dual-task performance is the key to solving this problem. Scalp acupuncture is a non-drug intervention method of traditional Chinese medicine to treat brain-derived diseases by acupuncturing the corresponding projection area of cerebral cortex function on the scalp. Studies have shown that scalp acupuncture helps improve neuronal damage and cognitive dysfunction and plays a neuroprotective function in central nervous system diseases. However, no relevant studies have discussed the synergistic gain effect of motor-cognitive dual task and scalp acupuncture. Therefore, this protocol aims to demonstrate the standardized operation of scalp acupuncture synchronizing motor-cognitive dual task and motor-cognitive dual task and compares the differences between these two tasks in healthy subjects through a randomized cross-over trial. This protocol initially revealed the possible influence mechanism of scalp acupuncture synchronizing motor-cognitive dual task on cognitive performance, gait control, and cortical brain function, which can provide new ideas and a theoretical basis for clinical exploration of new and effective non-drug treatment of integrated Chinese and Western medicine.

This protocol demonstrates the standardized procedure of scalp acupuncture synchronizing motor-cognitive dual task and motor-cognitive dual task. This can provide an important reference for the clinical exploration of the new and effective non-drug treatment of integrated Chinese and Western medicine.

Studies have shown that motor-cognitive dual task can greatly improve motor/cognitive function. However, the therapeutic effect of motor-cognitive dual ...

Transcranial Pulse Stimulation for Alzheimer's Patients

Transcranial pulse stimulation (TPS) is a noninvasive neuromodulation therapy with Conformit&#233; Europ&#233;enne (CE) marking for the treatment of Alzheimer's disease (AD). Initial pilot studies have demonstrated promising effects on cognitive function. This article focuses on the procedure for treating patients with AD using an MRI-guided, neuro-navigated TPS device. The protocol to be followed for this is described in detail, including the necessary procedures and device settings. A brief overview of the representative clinical results published to date is also provided. In addition to significant clinical improvements in cognition and affect, adverse events (AE) and possible adverse device events (ADE) are presented to provide safety data. Finally, the method is critically discussed. In the future, randomized controlled trials should be conducted to rule out any placebo effects. There is also currently a lack of long-term studies with a larger number of patients. Despite these unresolved questions, TPS has the potential as an adjunct treatment for Alzheimer's patients when used in a controlled, scientifically guided setting.

This article describes the procedure of transcranial pulse stimulation in patients with Alzheimer's disease. It discusses the indications, methodology, and future prospects in detail.

Transcranial pulse stimulation (TPS) is a noninvasive neuromodulation therapy with Conformit&#233; Europ&#233;enne (CE) marking for the treatment of ...

Acupoint Catgut Embedding for Chronic Pelvic Pain Management

Acupoint Catgut Embedding for Treatment of Chronic Pelvic Pain Due to the Sequelae of Pelvic Inflammatory Disease

Chronic pelvic pain caused by the sequelae of inflammatory pelvic disease is a common clinical condition of pelvic pain in women. At present, the main challenges in its treatment are the limited effectiveness of pain relief and the frequent recurrence of symptoms, which significantly impact patients' quality of life and impose a considerable psychological burden on them. It is a clinically challenging disease. After summarizing years of treatment experience, the author's team discovered that acupoint catgut embedding demonstrated notable clinical efficacy in managing chronic pelvic pain stemming from pelvic inflammatory disease sequelae. Compared to existing Western medicine treatment methods, acupoint catgut embedding offers advantages such as a good analgesic effect, lower recurrence rate, economic benefits, and a relatively straightforward procedure. This article provides a comprehensive guide on embedding absorbable catgut into patients' acupoints for the treatment of chronic pelvic pain in females resulting from the sequelae of pelvic inflammatory disease.

Author Spotlight: Enhancing Women's Chronic Pelvic Pain Management Through Acupoint Catgut Embedding

This article outlines acupoint catgut embedding therapy for chronic pelvic pain caused by the sequelae of pelvic inflammatory diseases.

Chronic pelvic pain caused by the sequelae of inflammatory pelvic disease is a common clinical condition of pelvic pain in women. At present, the main ...

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 ...

An Acupoint Catgut-embedding Therapy for Treating Obesity

Obesity is a chronic and complex disease caused by excessive fat deposition, which can lead to multiple complications. It affects individuals of all ages and backgrounds, contributing to a growing public health crisis worldwide. In modern medicine, the available treatments for obesity include drugs, behavioral modifications, and surgical procedures. However, their efficacy is often unsatisfactory and may be accompanied by various side effects. In contrast, Traditional Chinese Medicine offers a wide range of treatment options, including herbal tonics, acupuncture, tuina, acupoint catgut-embedding therapy, and more. Among these, acupoint catgut-embedding therapy has emerged as a popular choice for weight management due to its effectiveness, convenience, affordability, and minimal adverse effects. It involves the insertion of absorbable sutures into acupoints, stimulating the body's natural healing processes and enhancing metabolic function. The gradual absorption of the sutures over time provides a sustained therapeutic effect, allowing for prolonged benefits without the need for continuous treatment sessions. Whether used alone or in conjunction with other therapies, acupoint catgut-embedding therapy exemplifies the benefits of integrative medicine and its potential to empower individuals on their journey toward a healthier lifestyle. Aiming to visualize this ethnic medicine theory and promote its effective inheritance, we detail the acupoint catgut-embedding therapy method for weight loss, covering acupuncture points selection, materials, procedures, and precautions.

Here, we propose a specialized Chinese medicine treatment protocol to manage obesity. This treatment achieves weight loss by implanting absorbable surgical sutures into specific acupuncture points.

Obesity is a chronic and complex disease caused by excessive fat deposition, which can lead to multiple complications. It affects individuals of all ages ...