感谢陆军医科大学第二附属医院神经外科的现场支持。资助：本研究由中国自然科学基金（82104696）资助。

本研究的实验符合动物伦理学的“3R”原则，并已获得陆军医科大学实验动物福利和民族委员会（AMUWEC20234543）的批准。
1.啮齿动物身体固定装置的制备
<ol><li>基于中国实用新型专利9设计一种小动物支架。</li>
	<li>通过3D打印技术生产设备。</li>
	<li>准备一根 2 m 长的绳子，分成 6 根绳子。将这 6 根绳索穿过固定器上的孔。利用弹簧加载扣固定绳索末端，便于调整环尺寸。 注意：固定器可以由任何配备基于设计图纸的 3D 打印技术的公司制造。该器件的设计图如 图1所示。本研究中使用的固定器是由 3D 打印机通过一种称为熔融沉积建模 （FDM） 的打印方法生产的，也称为熔融长丝制造 （FFF）10。这种增材制造方法通过选择性地将熔化形式的材料沉积在预定路径中，逐层打印设计的零件。该工艺使用长丝形式的热塑性聚合物来创造最终的物理结构。本研究中使用的固定装置可以同时固定三只动物，需要六根绳索，每只动物需要一对绳索：一根用于颈部，一根用于躯干。</li>
</ol>2.啮齿动物的制备
<ol><li>购买3只Sprague-Dawley大鼠（10-40g，7-45日龄）和3只C57 BL / C小鼠（5-40g）。</li>
	<li>在昼夜循环12小时，环境温度为21-23°C，湿度为45%-60%的房间内饲养动物。定期对饲养环境进行消毒通风，让动物自由进食、饮水。</li>
</ol>3. 将啮齿动物固定在固定器上（俯卧位）
<ol><li>将啮齿动物的头部放在固定器前面的U形孔中，使啮齿动物处于俯卧位。将啮齿动物的身体穿过两个绳环。</li>
	<li>将绳索系在啮齿动物的脖子和腹部，确保它不会逃脱，但不要太紧，以免引起呼吸窘迫和死亡。<ol><li>关键步骤是将颈部固定到位。在啮齿动物的脖子和绳子之间留出 2-5 毫米的间隙。确保绳子不会太紧而无法勒死动物或太松而无法逃脱。</li>
	</ol></li>
	<li>在设备中以相同的方式修复其他啮齿动物。</li>
	<li>重复上述固定步骤（3.1-3.2）3天，以尽量减少固定过程引起的潜在应激反应。连续 3 天重复固定程序，每次持续 5 分钟。 注意：为确保啮齿动物在固定过程中不会过度紧张，我们连续 3 天重复该过程。每次治疗每天持续 5 分钟，为小鼠或大鼠提供充足的时间来适应该程序并最大限度地减少任何潜在的不适。</li>
	<li>从第 4 天开始 Strat EA。</li>
	<li>为防止疾病传播，请将无菌棉签或棉球浸入 75% 的酒精中，并在每次使用前后彻底擦拭支架以清洁和消毒。处理啮齿动物时，请使用手套和其他防护设备，以避免潜在的健康风险。</li>
</ol>4. 在小鼠或年轻大鼠中执行 EA （MS6）
<ol><li>选择 0.25 mm x 13 mm 一次性无菌针灸针。</li>
	<li>按解剖位置找到穴位，并确认穴位处的插入深度。 注意： MS6 位于头部的外侧。头部的这个区域穿过神聪（Ex-hn1）和玄里（GB6）之间的连接，也称为前顶叶（Du21）。MS6对应于头皮上大脑皮层中央前回的表达。Ex-hn1 位于头部前后中线中点后 0.5 cm 处。GB6位于耳朵前部的中点和眼睛的末端角。滑车上上神经、眶上神经、枕神经和耳颞神经分布在其中。MS6主要用于治疗中风和脑病，如运动障碍。</li>
	<li>在3只啮齿动物中，从前申冲（Exhn1）到玄礼（GB6）沿皮下，分别在3/5深度处将双侧无菌针头拧入MS6。</li>
	<li>将连接电缆的一侧连接到 EA 仪器。</li>
	<li>将包含正负EA夹的连接电缆的另一侧连接到左右MS6上的针灸针柄。</li>
	<li>打开 EA 仪器。<ol><li>旋转按钮（功能/选择开关）设置频率和功能模式。</li>
		<li>将 频率 设置为 10 Hz，然后选择 连续波 模式。</li>
		<li>旋转相应的按钮（调整强度）以设置强度。每个调整强度按钮控制其下方连接的电线的啮齿动物的电流强度。将电流强度调整到啮齿动物可接受的范围，通常为 1-2 mA，持续 5 分钟。 注意： 在手术过程中监测啮齿动物的呼吸和行为非常重要，以确保它们不会感到任何痛苦或不适。如果大鼠似乎在挣扎或疼痛，则应立即停止该程序。</li>
	</ol></li>
</ol>5. 将啮齿动物固定在固定器上（仰卧位）
<ol><li>将啮齿动物的头部放在固定器前面的U形孔中，使啮齿动物处于俯卧位。将啮齿动物的身体穿过两个绳环。</li>
	<li>将绳子系在啮齿动物的脖子和腹部，确保它不会逃脱，但不要太紧以至于导致呼吸窘迫和死亡。 注意：固定过程中要注意的要点与步骤3.2相同。</li>
	<li>以同样的方式修复其他啮齿动物。</li>
	<li>与步骤3.4一样，重复上述固定步骤3天，以减少啮齿动物的应激反应。</li>
	<li>从第 4 天开始 Strat EA。 注意： 按照步骤 3.6 中所述进行消毒。</li>
</ol>6. 执行 EA （ST25）
<ol><li>选择 0.25 mm x 13 mm 一次性无菌针灸针。</li>
	<li>按解剖位置找到穴位，并确认穴位处的插入深度。 注意： ST25 位于脐部水平。在啮齿动物中定位这个点需要首先测量啮齿动物爪子的长度。ST25 位于肚脐旁边爪子长度的 2/3。例如，如果测量的啮齿动物爪子为 0.6 cm，则 ST25 位于脐部左右 0.4 cm。</li>
	<li>在3只啮齿动物上，分别以4-8mm的深度缓慢扭曲并穿透无菌针头进入ST25双侧。</li>
	<li>将连接电缆的一侧连接到 EA 仪器。</li>
	<li>将包含正负EA夹的连接电缆的另一侧连接到左右ST25的针灸针柄。</li>
	<li>打开 EA 仪器。<ol><li>旋转按钮（功能/选择开关）设置频率和功能模式。</li>
		<li>将 频率 设置为 10 Hz，然后选择 连续波模式。</li>
		<li>旋转相应的按钮（调整强度）以设置强度。每个调整强度按钮控制其下方连接的电线的老鼠的电流强度。将电流强度调整到啮齿动物可接受的范围，通常为 1-2 mA，持续 5 分钟。 注：观察结果与步骤4.6相同。</li>
	</ol></li>
</ol>

使用 3D 打印支架对小动物进行体电针

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提交人声明不存在相互竞争的利益冲突。

电针 （EA） 是一种针灸形式，涉及在针灸针上使用电刺激11。该技术涉及使用特定强度和频率的微脉冲电流来刺激穴位并达到增强的治疗效果3.然而，EA的愈合机制是有限的，需要大量的基础研究来证明1,2,3。啮齿动物模型通常用于基本的 EA 研究 8,12,13<...

电针疗法是一种独特的方法，将针灸针插入头皮并连接到电机以刺激特定穴位 1。与手动刺激不同，EA 通过稳定特定频率和波形来更好地控制刺激，以实现最佳治疗效果2。据调查，中国81.2%的医疗机构使用EA或手针灸治疗脑瘫、神经痛、面瘫等疾病。尽管很受欢迎，但EA的具体疗效机制仍然未知，这阻碍了其在神经系统疾病康复治疗中的推广和应用1,2,3。需要进一步的研究来充分了解EA的有效性机制，并促进其在神经系统疾病康复治疗中的应用。
随着针灸在世界范围内的影响力不断扩大，许多研究已经研究了对啮齿动物模型（例如大鼠或小鼠模型）进行 EA 的机制 1,2,3。在EA的啮齿动物实验中经常会遇到几个问题。首先是如何在没有麻醉的情况下固定啮齿动物，因为对清醒受试者进行的针灸更能反映临床实践。此外，一些实验要求动物保持清醒以观察治疗效果2,3。另一个挑战是准确定位小鼠或大鼠的穴位，这些穴位与人类的穴位相对应。目前，许多学者正在研究实验啮齿动物穴位的精确定位4,5。在该协议中，选择了MS6和ST25，它们已通过人体解剖定位的转换在啮齿动物中明确定义。MS6 通常用于治疗某些脑部疾病，例如帕金森病6。ST25 通常用于治疗胃肠道问题，例如腹泻7.选择这两个穴位主要是为了展示如何在仰卧位和俯卧位上有效地固定啮齿动物。此外，这些穴位已被广泛研究，并为研究目的提供了重要的见解 6,7。
以前固定一只大鼠进行实验的方法不仅耗时，而且很难由一个人处理 8.此外，由于动物的不合作，在实践中成功率相对较低。因此，迫切需要创建一个易于建立的具有稳定特性的动物模型，以提高实验效率。在这篇文章中，介绍了一种用于小动物的3D打印支架，可以很容易地固定多只啮齿动物，导致运动受限。本文的目的是对一批年轻大鼠或小鼠进行EA治疗，重点研究小鼠批量限制、识别和刺激MS6和ST25穴位的策略。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3D printing batch mice fixator</td>
			<td>MESH INVENT</td>
			<td>Custom made</td>
			<td>The fixator produced by 3D printer. The printing method is called fused deposition modeling (FDM), also known as fused filament fabrication (FFF).&#160;&#160;</td>
		</tr>
		<tr>
			<td>Electroacupuncture instrument</td>
			<td>Hwato, Suzhou Medical Appliance Factory</td>
			<td>SDZ-III</td>
			<td>www.Hwato-med.com</td>
		</tr>
		<tr>
			<td>Disposable sterile acupuncture needle</td>
			<td>Suzhou Medical Appliance Factory</td>
			<td>N/A</td>
			<td>0.25 mm x 13 mm</td>
		</tr>
	</tbody>
</table>

bulk electroacupuncture operation for mice or young rats

电针（EA）被广泛用于治疗各种健康状况。然而，EA治疗的潜在机制尚不清楚，阻碍了其推广。机理研究需要小鼠或大鼠模型来解决这个问题。然而，这些动物不服从实验过程，这很耗时。为了解决这些问题，我们设计了3D打印的小动物身体散装固定器，以提高EA动物实验的效率。该视频详细介绍了如何使用固定器对小鼠或年轻大鼠进行批量 EA。对于穴位的选择，选取顶点颞部（MS6头部）和天枢点（ST25腹部）的前斜线，验证俯卧位和仰卧位固定装置的效果。使用3D打印的小动物支架可以同时固定和治疗多个啮齿动物，从而减少实验所需的时间和资源。这种技术可以通过3D打印不同尺寸的动物模型应用于其他动物模型，并有可能用于各种固定条件。该设备有利于促进EA的实验科学研究。

Electroacupuncture (EA) therapy is a unique method in which acupuncture needles are inserted into the scalp and connected to an electro-machine to stimulate specific points1. Unlike manual stimulation, EA allows better control of stimulation by stabilizing specific frequencies and waveforms to achieve optimal therapeutic effects2. According to a survey, 81.2% of medical institutions in China use EA or manual acupuncture to treat cerebral palsy, neuralgia, facial palsy, and other conditions. Despite its popularity, the specific mechanism of efficacy of EA is still unknown, which has hindered its promotion and application in the rehabilitation treatment of neurological diseases1,2,3. Further research is needed to fully understand the mechanism of EA&#39;s effectiveness and to promote its use in the rehabilitation treatment of neurological diseases.
As the influence of acupuncture expands worldwide, many studies have already investigated the mechanisms of EA performed on rodent models, such as rat or mouse models1,2,3. Several issues are often encountered in EA&#39;s rodent experiments. The first is how to immobilize rodents without anesthesia, as acupuncture performed on awake subjects is more reflective of clinical practice. In addition, some experiments require animals to be awake to observe treatment effects2,3. Another challenge is accurately locating the acupoints in mice or rats that correspond to those in humans. The precise localization of acupoints in experimental rodents is currently being studied by many scholars4,5. In this protocol, MS6 and ST25 were selected, which have been clearly defined in rodents by the transformation of human anatomical localization. MS6 is commonly used for the treatment of some brain diseases, such as Parkinson&#39;s disease6. ST25 is usually used to treat gastrointestinal problems, such as diarrhea7. These two acupoints were chosen primarily to demonstrate how rodents can be effectively immobilized in both supine and prone positions. Moreover, these acupoints have been extensively studied and offer significant insights for research purposes6,7.
The previous method of immobilizing a single rat for experiments is not only time-consuming but also difficult to handle by a single person8. Additionally, due to the non-cooperation of animals, the success rate is relatively low in practice. Therefore, there is a critical need to create an easily established animal model with stable characteristics to improve experiment efficiency. In this article, a 3D printed holder for small animals was introduced that could easily immobilize multiple rodents, leading to motor restriction. The aim of this paper is to administer EA treatments to a batch of young rats or mice, focusing on the strategies for bulk restriction of mice, identification, and stimulation of MS6 and ST25 acupoints.

作者聚焦：使用 3D 打印支架同时进行多次电针手术

小鼠或幼鼠的批量电针操作

In this video, we introduce an efficient way of using electroacupuncture on multiple awake mice or rats. In vesical research, animal is urinating, but they are not cooperative, and acupuncture only is time consume. Another issue is about the anesthesia, which is not clinical applicable of acupuncture.To address these issues, we demonstrate efficient method of performing electroacupuncture on multiple awake mice or rats by using 3D printed holder. Our protocol demonstrates a more efficiency and safer way to hold multiple animals simultaneously, which unlike other method that imply immobilization, this method not only saves time, but also improve experimental success rates. Additionally, device is designed to be easy to handle, which could be operated by one person.The ProTech and device are beneficial for the promotion of experimental scientific research in electroacupuncture.

我们设计的动物身体固定器的平面图如图1所示。此外，还提交了该固定器的 3D 图形模型，以提供设计的全面视图（补充文件 1）。这是一种允许 3 只啮齿动物固定并同时执行 EA 的设备。大鼠和小鼠在清醒状态下被限制在固定器内，俯卧位和仰卧位都可以牢固地固定，而不会对啮齿动物造成伤害（图2和图3</strong...

Watch this Scientific Journal Video about Bulk Electroacupuncture Operation for Mice or Young Rats at JoVE.com

在这里，我们提出了一种使用三维（3D）打印支架在小鼠或年轻大鼠中进行高效和快速电针（EA）的方案。该技术允许在多只动物身上同时操作，节省时间并提高实验效率。

Electroacupuncture (EA) is widely used to treat various health conditions. However, the underlying mechanism of EA treatment remains unclear, hindering ...

在本视频中，我们介绍了一种在多只清醒的小鼠或大鼠身上使用电针的有效方法。在膀胱研究中，动物在排尿，但它们不合作，针灸只是消耗时间。另一个问题是关于麻醉，这在针灸的临床上不适用。为了解决这些问题，我们展示了使用3D打印支架对多只清醒的小鼠或大鼠进行电针灸的有效方法。我们的方案展示了一种更有效、更安全的方式来同时持有多只动物，这与其他意味着固定的方法不同，这种方法不仅节省了时间，而且提高了实验成功率。此外，该设备设计为易于操作，可由一个人操作。ProTech和设备有利于促进电针的实验科学研究。

bulk-electroacupuncture-operation-for-mice-or-young-rats

Research

JoVE Journal

Biology

Bulk Electroacupuncture Operation for Mice or Young Rats

442 次观看.  Army Medical University. 在本视频中，我们介绍了一种在多只清醒的小鼠或大鼠身上使用电针的有效方法。在膀胱研究中，动物在排尿，但它们不合作，针灸只是消耗时间。另一个问题是关于麻醉，这在针灸的临床上不适用。为了解决这些问题，我们展示了使用3D打印支架对多只清醒的小鼠或大鼠进行电针灸的有效方法。我们的方案展示了一种更有效、更安全的方式来同时持有多只动物，这与?...