作者感谢组织学的布赖恩·约翰逊和影像核心在华盛顿大学与三色染色和分析的帮助。这项工作是由美国国立卫生研究院资助HL098067和HL089455支持。

机构的动物护理和使用委员会（IACUC）在华盛顿和雪松 - 西奈医学中心大学的必要批准这些研究动物的工作。 1.准备<ol><li>既消毒钝端钳和降压通过高压灭菌。 </li><li>使用生物安全柜，从冻干粉末制备BLM的工作原液在PBS中。超声处理10分钟，该溶液在35千赫到保证均匀混合。 注:建议的30和45微升之间的总体积，以防止与较大体积在低端移液变异，和窒息。 </li><li>准备一个干净的工作区，其包括用于该过程本身约1米2，以及两个前和后的程序笼指定地点。 </li><li>通过在底座和underl铺设2或3条实验室胶带固定的程序董事会立即在研究者面前的板凳基地颖替补。 图 1所示为创造一个板进一步规范。 </li><li>领带的过程板的两个定位螺丝之间尺寸4.0缝合线的单根长度。 </li><li>产生通过拆卸和从三个1毫升注射器丢弃柱塞，以及沉积60微升PBS中到每个筒的顶部上以形成气密密封的一个临时的肺活量计。固定导管的枢纽松散到注射器之一，它放置在电路板的一侧。 </li><li>吸300微升空气进入1毫升的注射器，并将其放置在电路板的一侧。 </li><li>切割胶带在长度和地点约6英寸的附加片一侧。这将被用于动物固定在板步骤2.4。 </li><li>设置一个异氟醚室。附加O 2，异氟醚，真空到相应的端口上都曝光室和真空间隙。另外，施用麻醉剂在异氟醚兼容的生物安全柜。 </li></ol> 2.插管<ol><li>麻醉异氟烷鼠标在室内，直到它失去意识，呼吸减慢，以适当的速度。典型的曝光包括3至4分钟的4％的异氟醚和2％O 2，和理想的结果是2〜2.5分钟的镇静。这对应于1呼吸每2秒的呼吸速率。 </li><li>在等待镇静设置在，吸BLM的30和45微升之间成移液器和地点到一侧。 </li><li>当准备好后，从连接于程序平台的定位螺钉的螺纹暂停其上门牙的镇静鼠标。确保动物的背部靠在台表面平坦。 </li><li>小心不要限制通风，放置一块胶布松散整个胸腔下部（尾）部分，只是隔膜上方。位置应足够紧PROC过程中保持正确对准edure，但不是那么紧，它限制呼吸。 </li><li>打开照明灯％和100％的强度80之间和定向鹅颈，以便它是从皮肤的表面1至2厘米，靠近太阳穴。定期检查鹅颈管的尖端热，以确保它不伤害老鼠。 </li><li>站在平台背后，用无菌，钝端钳定位舌头。小心，以避免下切牙，轻轻夹住并绘制舌头口腔。 </li><li>使用剩余的手，插入抑制剂并用它来压平舌针对口腔的楼层。松开钳，但留下的降压到位接下来的两个步骤。 </li><li>定向光使气管是由从太阳穴的水平向近侧引导鹅颈管，直到它到达主支气管的水平可见的。 注意:气管可通过呼吸的作用而容易地分开，WHICH导致发出的光的强度波动。当正确定位时，该结构将在轴向平面内的光的一个位于中心的销与在口腔中本身最小环境光可辨别。 </li><li>角的注射器，使得它跟随气管的自然路径，并降低22-G的导管末端，与含有微滴，直入管腔所附注射器。 PBS的泡沫将开始上升，并在成功安置每次呼吸下降。 注意:此动作可通过几秒钟作为深度镇静的结果被延迟。 </li><li>喂导管在额外的5毫米。取出压舌板。 </li><li>移注射器相反的手，抓住中心，轻轻取下注射器。 </li><li> BLM的30和45微升之间存到导管毂的内部的中心，连接第二注射器和分配300μl的空气进入中心。 </li><li>更换第二与第一含PBS的气泡注射器。泡沫将继续上升，如果程序已经成功地进行下降。 </li></ol> 3.后护理程序<ol><li>取下导管和胶带，然后将动物在干燥温暖的地方，直到它恢复意识 - 通常在一两分钟。 </li></ol>

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The authors have nothing to disclose.

在雾化是不切实际由于有限的试剂可用性，安全性或成本的情况下，直接气管施用是用于递送外源剂的进入肺部优良的方法16经气管滴注已被广泛用于完成此。然而，与所有的外科手术，它也与它携带用于引起该过程本身并发症的可能性，并且不一定剂被灌输。13 由于这些原因，已经变得越来越普遍，直接经由插管16,17,18,19,20-服用一种物质进入气管。然而，?...

尽管某些解剖和生理差异，1鼠模型继续是无价建模人类生物学和疾病的发病机制。2从饲养的角度来看，小鼠是容易处理，具有低的繁殖时间，加速寿命，并且是相对便宜的去家。用不同遗传品系和策略（例如，条件敲奏，记者小鼠谱系追踪方法等），以及在宽范围内可用的试剂（例如，抗体，重组蛋白，抑制剂的开发，等），老鼠已经成为一个重要的脊椎动物模型生物体揭示人类的稳态和疾病过程。3 小鼠已经特别有价值用于研究肺部疾病，包括急性肺损伤（ALI）和肺纤维化。4 ALI在人类可以通过创伤，损伤或败血症引起，其特征是上皮和内皮泄漏（即，水肿），炎症，和新生的纤维化。在许多患者中，急性肺损伤前进到它的严重的形式，急性呼吸窘迫综合征（ARDS），这经常导致纤维化和死亡因呼吸衰竭。5,6-肺纤维化是一个渐进的，致命的病理学特征在于细胞外基质的过量沉积，最显着的I型胶原，导致肺功能受损。-7,8-管理博莱霉素（BLM）是最广泛使用和最佳表征模型实验动物诱导ALI和纤维化。9虽然博莱霉素诱导的肺纤维化啮齿动物确实不概括完全人类纤维化表型，与该模型10小鼠研究已导致的影响的发生和疾病的进展许多重要因素的发现。11 虽然确切机制（S）的后面博莱霉素诱导的纤维化是未知的，则发起伤被认为是从在上皮细胞接触依赖性的DNA链断裂衬传导气道和肺泡，并且特别地，1型肺泡产生。12的必要性BLM和肺上皮之间的直接接触突出一个健壮的递送途径的重要性和这些问题也有密切关系广泛的治疗靶向远端气道，包括重组蛋白，抗体，siRNA的，病毒，细菌，微粒，等等。口咽抽吸（OPA）已被广泛地用于此目的13，但OPA的一个主要缺点是，递送剂的某些部分可以被吞咽到胃肠道，从而导致不精确的给药剂量。另一广泛使用的方法是经气管滴注，涉及强麻醉下气管到代理的气管和滴注暴露直接进入呼吸道。14然而，不仅可以这样一个过程是不希望的，由于它的invasivity，但它也是耗时的，需要的培训公平位，同时产生强效伤到呼吸道15,16若干协议已被开发涉及的试剂的直接施用进而不需要手术干预气管，引起强大麻醉药16,17,18,19,20-但这些方法包括延长的恢复时间，使用昂贵的设备（即，耳镜/喉镜，市售程序板，光纤线等），在口腔中的过量操纵，和不确定性有关的剂量。 本文描述的给药通过插管相对容易的方法，其允许研究者快速，廉价和可靠地灌输试剂进鼠肺与对周围组织的残余破坏的风险有限。

<table border="0" cellpadding="0" cellspacing="0" width="1528">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:481px;">Bleomycin For Injection, 30 units/vial</td>
			<td style="width:304px;">APP Pharmaceuticals, LLC</td>
			<td style="width:139px;">103720</td>
			<td style="width:604px;">For best results, BLM should be suspended in PBS, aliquoted, and stored as single use lyophilzed aliquots</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Blunt End Forceps</td>
			<td style="width:304px;">N/A</td>
			<td style="width:139px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tongue Depressor (i.e. bent Valleylab Blade Electrode, 2.4&#34;)&#160;</td>
			<td style="width:304px;">Covidien</td>
			<td>E1551G</td>
			<td>Before use, create a 45 degree bend 1.5 cm&#160; from the blade tip. A suitable depressor can also be created from any metal implement of similar dimensions.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Exel Safelet Catheter 22G X 1&#34;</td>
			<td style="width:304px;">Exel International</td>
			<td>26746</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1 mL Slip-tip Disposable Tuberculin Syringe (200/sp, 1600/ca)</td>
			<td style="width:304px;">BD</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.2ml Pipettor and Filter Tips</td>
			<td style="width:304px;">N/A</td>
			<td style="width:139px;">N/A</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:481px;">Fiber-Lite Illuminator. Model 181-1: Model 180 mated with Standard Dual Gooseneck illuminator</td>
			<td style="width:304px;">Dolan Jenner Industries, Inc.</td>
			<td style="width:139px;">181-1</td>
			<td>Lower output LED illuminators are not recommended as they fail to suficiently illuminate the trachea.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Intubation Board</td>
			<td style="width:304px;">N/A</td>
			<td style="width:139px;">N/A</td>
			<td>See Diagram 1.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Colored Label Tape: 0.5&#160;in. Wide</td>
			<td style="width:304px;">Fisherbrand</td>
			<td>15-901-15A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oxygen&#160;</td>
			<td style="width:304px;">N/A</td>
			<td style="width:139px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Phosphate-Buffered Saline, 1X</td>
			<td style="width:304px;">Corning</td>
			<td>21-040-CV</td>
			<td>Product should be sterile</td>
		</tr>
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			<td height="21" style="height:21px;">Non-Sterile Silk Black Braided Suture Spool, 91.4 m, Size 4-0</td>
			<td style="width:304px;">Harvard Apparatus</td>
			<td>517698</td>
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			<td height="21" style="height:21px;">Table Top Anesthesia Machine Isoflurane</td>
			<td style="width:304px;">Highland Medical Equipment</td>
			<td>N/A</td>
			<td>http://www.highlandmedical.net/</td>
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			<td height="21" style="height:21px;">Slide Top Induction Mouse Isoflurane Chamber</td>
			<td>MIP / Anesthesia Technologies</td>
			<td>AS-01-0530-SM</td>
			<td></td>
		</tr>
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			<td height="21" style="height:21px;">FORANE (isoflurane, USP) Liquid For Inhalation 100 mL&#160;</td>
			<td style="width:304px;">Baxter</td>
			<td>1001936040</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;Nanozoomer Digital Pathology system</td>
			<td style="width:304px;">Hamamatsu</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">IgM ELISA Quantification Kit&#160;</td>
			<td style="width:304px;">Bethyl Laboratories</td>
			<td style="width:139px;">E90-101</td>
			<td></td>
		</tr>
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an improved method for rapid intubation of the trachea in mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

如别处所述21插管小鼠每天监测体重减轻和窘迫，处死4,10或17天后通过2.5％2,2,2- 三溴腹腔注射。 支气管肺泡灌洗 （BAL）收集在PBS中洗涤三次和右肺固定在10％福尔马林，石蜡包埋，并用Masson三色由华盛顿组织学和成像核心22的大学染色。 在既定的数据一致，博莱霉素处理的小鼠经历7和10天暴露后<...

Watch this Scientific Journal Video about An Improved Method for Rapid Intubation of the Trachea in Mice at JoVE.com

An Improved Method for Rapid Intubation of the Trachea in Mice

在小鼠气管插管快速的改进方法

本文介绍了通过插管直接施用博莱到小鼠气管一种快速，简便的方法。这种方法的主要优点是，它是高度可再现的，容易掌握，并且不需要专门的设备或冗长的恢复时间。

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

an-improved-method-for-rapid-intubation-of-the-trachea-in-mice

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

25.0K 次观看.  University of Washington. 本文介绍了通过插管直接施用博莱到小鼠气管一种快速，简便的方法。这种方法的主要优点是，它是高度可再现的，容易掌握，并且不需要专门的设备或冗长的恢复时间。 

视频: An Improved Method for Rapid Intubation of the Trachea in Mice

Guidelines for Elective Pediatric Fiberoptic Intubation

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small babies will desaturate very quickly if ventilation is interrupted mainly to high metabolic rate. We describe guidelines to perform a safe fiberoptic intubation while maintaining spontaneous breathing throughout the procedure. Steps requiring the use of propofol pump, fentanyl, glycopyrrolate, red rubber catheter, metal insuflation hook, afrin, lubricant and lidocaine spray are shown.

We describe guidelines to perform a safe and efficient elective fiberoptic intubation in pediatric patients while maintaining spontaneous ventilation.

选修儿科光纤插管指南

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small ...

科研

医学

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

原位主动脉瓣移植在小鼠血管疾病的研究

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

Mice, both wildtype and transgenic, are the principal mammalian model in biomedical research currently. Intubation and mechanical ventilation are necessary for whole animal experiments that require surgery under deep anesthesia or measurements of lung function. Tracheostomy has been the standard for intubating the airway in these mice to allow mechanical ventilation. Orotracheal intubation has been reported but has not been successfully used in many studies because of the substantial technical difficulty or a requirement for highly specialized and expensive equipment. Here we report a technique of direct laryngoscopy using an otoscope fitted with a 2.0 mm speculum and using a 20 G&#160;intravenous catheter as an endotracheal tube. We have used this technique extensively and reliably to intubate and conduct accurate assessments of lung function in mice. This technique has proven safe, with essentially no animal loss in experienced hands. Moreover, this technique can be used for repeated studies of mice in chronic models.

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

We have developed a simple, reliable, and relatively inexpensive method for endotracheal intubation in mice via direct laryngoscopy using an otoscope with a 2.0 mm speculum. This technique is atraumatic and can be used for repeated measurements in chronic experiments. We find it superior to tracheostomy or previously reported nonsurgical techniques.

气管插管小鼠<em&gt;通过</em&gt;直接喉镜使用耳镜

Mice, both wildtype and transgenic, are the principal mammalian model in biomedical research currently. Intubation and mechanical ventilation are ...

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

一个简单的方法小鼠肺插管

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) models based on pBECs in mono-culture (MC), MC with astrocyte-conditioned medium (ACM), and non-contact co-culture (NCC) with astrocytes of porcine or rat origin. pBECs were isolated and cultured from fragments of capillaries from the brain cortices of domestic pigs 5-6 months old. These fragments were purified by careful removal of meninges, isolation and homogenization of grey matter, filtration, enzymatic digestion, and centrifugation. To further eliminate contaminating cells, the capillary fragments were cultured with puromycin-containing medium. When 60-95% confluent, pBECs growing from the capillary fragments were passaged to permeable membrane filter inserts and established in the models. To increase barrier tightness and BBB characteristic phenotype of pBECs, the cells were treated with the following differentiation factors: membrane permeant 8-CPT-cAMP (here abbreviated cAMP), hydrocortisone, and a phosphodiesterase inhibitor, RO-20-1724 (RO). The procedure was carried out over a period of 9-11 days, and when establishing the NCC model, the astrocytes were cultured 2-8 weeks in advance. Adherence to the described procedures in the protocol has allowed the establishment of endothelial layers with highly restricted paracellular permeability, with the NCC model showing an average transendothelial electrical resistance (TEER) of 1249 &#177; 80 &#937; cm2, and paracellular permeability (Papp) for Lucifer Yellow of 0.90 10-6 &#177; 0.13 10-6 cm sec-1 (mean &#177; SEM, n=55). Further evaluation of this pBEC phenotype showed good expression of the tight junctional proteins claudin 5, ZO-1, occludin and adherens junction protein p120 catenin. The model presented can be used for a range of studies of the BBB in health and disease and, with the highly restrictive paracellular permeability, this model is suitable for studies of transport and intracellular trafficking.

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of the protocol is to present an optimized procedure for the establishment of an in vitro blood-brain barrier (BBB) model based on primary porcine brain endothelial cells (pBECs). The model shows high reproducibility, high tightness, and is suitable for studies of transport and intracellular trafficking in drug discovery.

一种基于猪脑内皮细胞的体外血脑屏障模型的改进方法

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) ...

Assessment and Characterization of Hyaloid Vessels in Mice

In the eye, the embryonic hyaloid vessels nourish the developing lens and retina and regress when the retinal vessels develop. Persistent or failed regression of hyaloid vessels can be seen in diseases such as persistent hyperplastic primary vitreous (PHPV), leading to an obstructed light path and impaired visual function. Understanding the mechanisms underlying the hyaloid vessel regression may lead to new molecular insights into the vascular regression process and potential new ways to manage diseases with persistent hyaloid vessels. Here we describe the procedures for imaging hyaloid in live mice with optical coherence tomography (OCT) and fundus fluorescein angiography (FFA) and a detailed technical protocol of isolating and flat-mounting hyaloid ex vivo for quantitative analysis. Low-density lipoprotein receptor-related protein 5 (LRP5) knockout mice were used as an experimental model of persistent hyaloid vessels, to illustrate the techniques. Together, these techniques may facilitate a thorough assessment of hyaloid vessels as an experimental model of vascular regression and studies on the mechanism of persistent hyaloid vessels.

This protocol describes both in vivo and ex vivo methods to fully visualize and characterize hyaloid vessels, a model of vascular regression in mouse eyes, using optical coherence tomography and fundus fluorescein angiography for the live imaging and ex vivo isolation and subsequent flat mount of hyaloid for quantitative analysis.

小鼠透明血管的评估与表征

In the eye, the embryonic hyaloid vessels nourish the developing lens and retina and regress when the retinal vessels develop. Persistent or failed ...

A Revised Method for Inducing Secondary Lymphedema in the Hindlimb of Mice

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore potential treatment options. This mouse model allows researchers to induce significant lymphedema lasting at least 8 weeks. Lymphedema is induced using a combination of fractioned radiotherapy and surgical ablation of lymphatics. This model requires that the mice get a dose of 10 Gray (Gy) radiation before and after surgery. The surgical part of the model involves ligation of three lymph vessels and extraction of two lymph nodes from the mouse hindlimb. Having access to microsurgical tools and a microscope is essential, due to the small anatomical structures of mice. The advantage of this model is that it results in statistically significant lymphedema, which provides a good basis for evaluating different treatment options. It is also a great and easily available option for microsurgical training. The limitation of this model is that the procedure can be time consuming, especially if not practiced in advance. The model results in objectively quantifiable lymphedema in mice, without causing severe morbidity and has been tested in three separate projects.

This animal model enables researchers to induce statistically significant secondary lymphedema in the hindlimb of mice, lasting at least 8 weeks. The model can be used to study the pathophysiology of lymphedema and to investigate novel treatment options.

小鼠下肢诱导继发性淋巴水肿的修订方法

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore ...

Repeated Orotracheal Intubation in Mice

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ventral neck for direct confirmation of cannula placement in the trachea. The relative difficulty or the tissue trauma induced to the subject by such procedures can be an impediment to an investigator&#8217;s ability to perform longitudinal studies. This article illustrates a technique in which physical manipulation of the mouse following the use of a depilatory to remove hair from the ventral neck permits transcutaneous visualization of the trachea for orotracheal intubation regardless of degree of skin pigmentation. This method is innocuous to the subject and easily achieved with a limited understanding of murine anatomy. This refined approach facilitates repeated intubation, which may be necessary for monitoring progression of disease or instillation of treatments. Using this method may result in a reduction of the number of animals and technical skill required to measure lung function in mouse models of respiratory disease.

The goal of this article is to describe a refined method of intubation of the laboratory mouse. The method is noninvasive and, therefore, ideal for studies that require serial monitoring of respiratory function and/or instillation of treatments into the lung.

小鼠重复的奥内切管插管

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ...

Semi-Minimal Invasive Method to Induce Myocardial Infarction in Rats and the Assessment of Cardiac Function by an Isolated Working Heart System

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily and highly reproducible MI induction procedure is required to obtain further insight and better understanding of the underlying pathological changes. This procedure can also be used to evaluate the effects or potency of new and promising treatments (as drugs or interventions) in acute MI, subsequent remodeling and heart failure (HF). After intubation and pre-operative preparation of the animal, an anesthetic protocol with isoflurane was performed, and the surgical procedure was conducted&#160;quickly. Using a minimally invasive approach, the left anterior descending artery (LAD) was located and occluded by a ligature. The occlusion can be performed acutely for subsequent reperfusion (ischemia/reperfusion injury). Alternatively, the vessel can be ligated permanently to investigate the development of chronic MI, remodeling or HF. Despite common pitfalls, the drop-out rates are minimal. Various treatments such as remote ischemic conditioning can be examined for their cardioprotective potential pre-, peri- and post-operatively. The post-operative recovery was quick as the anesthesia was precisely controlled and the duration of the operation was short. Post-operative analgesia was administered for three days. The minimally invasive procedure reduces the risk of infection and inflammation. Furthermore, it facilitates rapid recovery. The &#8220;working heart&#8221; measurements were performed ex vivo and enabled precise control of preload, afterload and flow. This procedure requires specific equipment and training for adequate performance. This manuscript provides a detailed step-by-step introduction for conducting these measurements.

This article presents an efficient method to perform myocardial ischemia and subsequent chronic reperfusion in rats using a minimally invasive approach. In addition, left ventricular hemodynamic function of rats is assessed by echocardiography and isolated working heart methods.&#160;

诱导大鼠心肌梗死的半微创方法及独立工作心脏系统对心脏功能的评估

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily ...

A Modified Simple Method for Induction of Myocardial Infarction in Mice

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI remodeling and evaluation of novel therapeutics. Different methods (e.g., isoproterenol treatment, cryoinjury, coronary artery ligation, etc.) have been used to induce MI. Compared with isoproterenol treatment and cryoinjury, coronary artery ligation may better reflect the ischemic response and chronic remodeling after MI. However, traditional methods for coronary ligation in mice are technically challenging. The current study describes a simple and efficient process for induction of MI in mice with readily available materials. The mouse chest skin was cut open under stable anesthesia. The heart was immediately externalized through the intercostal space after blunt separation of the pectoralis major and pectoralis minor. The left anterior descending branch (LAD) was ligated with a 6-0 suture 3 mm from its origin. Following LAD ligation, staining with 2,3,5-Triphenyltetrazolium chloride (TTC) indicated successful induction of MI and temporal changes of post-MI scar size. Meanwhile, survival analysis results showed overt mortality within 7 days after MI, mainly due to cardiac rupture. Moreover, post-MI echocardiographic assessment demonstrated successful induction of contractile dysfunction and ventricular remodeling. Once mastered, an MI model can be established in mice within 2-3 min with readily available materials.

Under adequate anesthesia, the mouse heart was externalized through the intercostal space, and myocardial infarction was successfully induced by ligating the left anterior descending artery (LAD) using materials readily available in most laboratories.

一种改良的小鼠心肌梗死诱导简易方法

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI ...