The author is grateful for the assistance of Mrs. Sharon Spana (University of Sydney) in rodent monitoring, handling and experimentation. The authorwould like to thank Prof. Jonathan Stone and Dr. Dan Johnstone for the assistance in the preparation of this manuscript. PhD funding support has been provided by University of Sydney and Australian Center for Excellence in Vision.

伦理声明：本协议遵循悉尼大学的动物护理准则，AEC＃5657。麻醉是经动物伦理委员会（悉尼大学，AEC＃5657）。 1，设备的准备<ol><li>使用实时皮肤温度跟踪。打开电脑和数据采集硬件。 </li><li>打开温度记录软件，调整设定温度之间30-35℃和采样，以每100毫秒的频率。 </li><li>可选：插入直肠体温计，以保证核心温度保持稳定在37.5℃。 </li></ol> 2.校准手册血压计<ol><li>新生儿连接手臂袖口sphygmomanometer.Use大小2袖口为250-550克的老鼠。适配器可能需要袖带管路连接到血压计。 </li><li>由任一松开空气释放阀或从适应断开袖带管袖带放气要么。确保无压力保持在管中及压力计针搁置在零的椭圆形/矩形内。 </li><li>检查油管，压力计和袖带之间的压力。充气的充气球的温柔泵袖口，直到它读取100毫米汞柱的压力表。确保压力保持不变。慢慢打开空气释放值袖带放气。 </li></ol> 3.动物的制备注意：动物是经过光损伤需要暗适应远程缺血前一天晚上。动物进行光损伤需要暗饲养（12小时光照：黑暗周期（5勒克斯）） <ol><li>在任何醒着还是anesthetised啮齿类动物进行远程缺血。保证动物有一个健康的肌张力。按捏上后肢，以确认有足够的肌肉存在确保这一点。 RIP诱导保护，防止光损伤已在大鼠久坐经测试长达6个月的年龄。 </li><li>对于R Anesthetised准备IP <ol><li>注入大鼠腹膜内注射60毫克/千克氯胺酮和5mg / kg的甲苯噻嗪。通过延伸腿和捏在脚的下侧的皮肤检查麻醉深度。动物有没有反射，如果它深深地anesthetised。应用人工泪液，以免角膜干燥，而在麻醉下。 </li><li>放置大鼠在任一热垫或循环水加热器管保持37.5℃的恒定体温。定位大鼠与下肢&#39;脚垫朝上俯卧位。右侧或左肢体可经历远程缺血。 </li></ol></li><li>对于RIP清醒的准备 注：清醒动物实验需要两个人。一个人抑制了动物和第二个人操作手动血压计。实验者必须有信心来执行的程序，约束会增加受伤的处理程序的风险。接受远程缺血大鼠必须牛逼空调o手动克制。取决于直觉准则手册约束应该从30秒进展到最多5分钟在数星期。胆小的动物，无法适应环境约束手动应排除清醒实验。最后，手动约束很可能引起应力（和潜在介绍的困惑的研究）对动物和假队列（袖带放置无炎症）必须被用来准确地解释的RIP的研究结果。 <ol><li>切一块毛巾到15厘米×30〜50厘米长，然后将短边垂直于老鼠的脊椎，覆盖头部后肢的顶部。 </li><li>掖短边下老鼠的躯干紧紧地开始缠上毛巾的剩余长边的老鼠。固定臂下的包裹动物以仰卧姿势。如果老鼠是根据左臂举行，不受毛巾老鼠的右侧肢体。 </li></ol></li></ol> 4.应用Ø˚F皮肤温度探头<ol><li>延伸是经受缺血和放置在皮肤探头放在脚掌的大鼠的腿。定位皮肤探头最大化温度探头和皮肤之间的接触。牢固地将探头插入脚垫并加盖探头纸带。 </li><li>通过跟踪温度记录软件温度检查皮肤探针放置。确保皮肤温度是30-34之间℃，并保持稳定。跟踪皮肤温度1-2分钟。调整皮肤探头如果温度不稳定或低于30℃。 </li></ol> 5.远程缺血<ol><li>袖带放气，并保证空气压力阀关闭。伸腿和松散包围在上后肢的袖口。用食指和拇指伸腿和低位保持松软的袖口位置。 </li><li>提高anesthetised动物的袖带压力为160毫米汞柱，而在清醒动物incre酶袖带压力180毫米汞柱。 注：大鼠的anesthetised血压范围从120-140毫米汞柱，并上升到160毫米汞柱时意识。开始的计时器和脚的温度记录一次正确的压力为止。 注：脚的温度应在5分钟的恒压后2℃下降。 </li><li>保持上述动物的“拐点”在整个缺血​​箍的位置。袖带压力将开始几分钟后丢弃，或者如果老鼠的肢体运动。 </li><li>反复泵送充气球在短脉冲以维持期望的袖带压的重复短脉冲抽</li><li>远程缺血可连续进行5至15分钟，被传递。缺血再灌注协议包括2中间有一个5分钟再灌注期5分钟缺血。 </li><li>通过松开空气压力阀袖带放气压力。检查温度变化过缺血亲的过程母育。松开袖带。 </li><li>继续损伤的实验。在麻醉状态下的效果的动物将需要被放置在加热垫上。继续监测动物，直到门诊。动物不能退回住房，直到行走。 </li></ol> 6.光损伤 - 视网膜变性模型<ol><li>暗适应动物过夜（12-15小时）。紧接着远程缺血空调或假远程缺血（动物约束）的地方动物有机玻璃外壳，食物和水。 </li><li>打开上方的有机玻璃外壳在上午9点24小时荧光灯（1000勒克斯）。曝光后，返回动物昏暗灯光循环7天。 </li></ol> 7.后远程缺血程序<ol><li>视觉评估与电图（ERG）： 注：ERG设置和闪光灯协议遵循布兰德利和石26。 <ol><li>一夜之间暗适应动物（12-15小时）。下暗淡红色照明通过腹膜内注射氯胺酮和赛拉嗪（60毫克/公斤和5毫克/公斤，分别）麻醉动物。散瞳（硫酸阿托品1.0％），角膜麻醉剂（丙美卡因0.5％）。 </li><li>应用角膜水化（卡波姆聚合物）的眼睛立即下降到角膜。适用于眼部凝胶，在20分钟的间隔，以维持角膜的​​水化。 </li><li>周围画一个眼球捆绑松散线程，以帮助稳定ERG录音。用直肠探头监测温度，保持动物的体温在37-37.5℃。 </li><li>放置一个Ganzfeld积分球内的头部。 注：Ganzfeld是一个完全可编程的光刺激，可提供统一的白衣闪烁的LED从的眼睛。 </li><li>记录使用定做4毫米铂正极轻轻触摸角膜和一个直径为2毫米的Ag / AgCl颗粒电极插入口的电图。参考两个电极的不锈钢针插入皮下臀部。 </li><li>与0.3-1,000赫兹（-3分贝）带通设置记录的信号，以2 kHz的采集速率（公元仪器）。经过一个稳定的ERG记录建立主题的动物有10分钟的暗适应录音开始前。 </li><li>按照先前由布兰德利和石26中描述的闪光灯协议。 <ol><li>计划闪光的持续时间（我们使用闪烁1-2毫秒的持续时间），并设置其强度为-4.4至2.0日志苏格兰人cd.sm -2。使用明亮的闪烁（2.0日志苏格兰人cd.sm -2，1毫秒）来衡量视网膜功能。在这项研究中是控制，光损伤和光损伤RIP之间的比较。 </li></ol></li></ol></li></ol> 8. TUNEL法<ol><li>通过腹膜内注射苯巴比妥过量（100毫克/千克）的动物实施安乐死。剜除眼睛和4％多聚甲醛固定。 </li><li> cyroprotecting前PBS洗眼睛眼睛过夜蔗糖30％（重量/体积）。嵌入眼睛在OCT化合物中并切成使用低温恒温器20微米的矢状切面。 </li><li>以下Maslim 等人的协议来执行与DAPI染色的视网膜切片的TUNEL法。27 </li><li>使用荧光显微镜的视网膜TUNEL计数。 TUNEL染色的细胞从外核层（ONL）记录;其中包含感光细胞核视网膜的最外层。在这项研究中，TUNEL计数一式三份作了为每只眼睛，用5眼对于每个处理组。 </li><li>使用单因素方差分析统计比较组是指控制，光损伤和光损伤+ RIP只。 </li></ol>

The author has nothing to disclose.

啮齿动物后肢缺血成功诱导带有手动血压计和袖口递送神经保护视网膜的光感受器。一发现与光损伤9,28缺血诱导调节感光保护一致。 从本质上讲，远程缺血引起短暂的缺氧给组织。因此，远程缺血预处理有许多相似之处缺血空调或交替称为缺血耐受，缺氧预处理，并在一定程度上，无氧运动。细胞缺血挑战通过释放大量的各种蛋白质，核苷和转录因子直接提供神经?...

大多数存活，也许所有，在代谢应激的面组织可以通过现有调理用周期亚致死缺血1,2的改进。缺血预处理（IP）在实际应用中是组织的暴露于亚致死缺血，组织经历更严重的压力，如随后的缺血性损伤之前。在动物模型中，IP提供给大脑，视网膜，心脏和肺3-6醒目的保护。与此相对应，在中风患者观察表明先前短暂性缺血发作和更好的临床结果7,8之间的链接。 IP也保护非缺血性损伤9视网膜感光。 知识产权在不同组织和伤害效果表明，它是活化细胞存活存在于所有组织的固有机制。心肌缺血预处理已建议有通过上调保护作用缺氧诱导因子（HIF），已知通过腺苷的释放或通过线粒体ATP钾通道10,11的开口，以调节许多代谢途径。腺苷释放和ATP钾通道有牵连的脑缺血但，调查缺血调理迄今为止的神经保护机制已经被集中在修改反兴奋性毒性，抗凋亡和抗炎通路12,13。总体上，缺血性调理用于保护神经元的分子过程的理解是有限的。 远程缺血预处理试图调节遥远极为重要器官（心脏，脑，肺）通过在较不关键组织产生缺血。使用后肢远程缺血预处理（RIP）已被证明是神经保护性中风14-17的啮齿动物模型。由我们所描述的方法提供了一种简单，可靠和非侵入性PROTocol诱导RIP。 RIP的协议绝大多数涉及后肢，大概是因为股动脉位于上后肢可以很容易地识别和手术夹紧和止血带的应用程序访问。在大脑和皮肤保护的研究侵入性肢体缺血的研究，缺血是由分离腹股沟韧带的股动脉夹闭股动脉致2,15,18。 从任一肢袖套或股动脉夹紧造成的缺血已证实在​​肢体变化，包括损失脉冲，降低氧合和皮肤温度下降。远程缺血可通过脉冲通过使用激光多普勒或超声多普勒17-19损失来确认。皮肤温度可以用作替代多普勒尽管这种关系是非线性的20,21。精确的温度记录是司空见惯的实验室和CAN很容易地纳入远程缺血性研究。 一种替代股骨夹紧手术是缺血使用止血带的诱导。止血带的应用程序产生媲美缺血到实现了与船舶夹紧; Kutchner 等 。相比侵入股动脉夹紧到一个非侵入止血带，发现两种方法停止在皮瓣缺血18整形外科模型血液流向肢体和减少皮肤损伤。掴或腿或臂和提高袖带压力到高于收缩期血压已发现可预防在猪和人类17,19,22局部缺血损伤。 不同止血带接近诱发缺血性远程包括使用血压袖带或松紧带17,22,23的。然而，使用弹性带以诱发局部缺血是一种不安全的方法，有可能引起的压力未调节量在肢，与压力上升到高于500毫米汞柱被记录在人类24。另外，使用弹性带肢体缺血导致肌肉损伤后大鼠除去带23的，如评估通过伊文思蓝染料， 在体内标记的肌纤维渗透率25。与此相反，输送一个受控的压力向止血带可以使用连接到一个血压计17,19,22,26血压袖带来实现。 在这项研究中，光感受器变性的光损伤模型，用于演示的远程缺血预处理的神经保护功效。远程缺血立即被诱导之前光损伤，并阻止随后的光感受器变性证实了视网膜功能测试。随行的视频将展示非侵入性的远程缺血的应用。

<table border="0" cellpadding="0" cellspacing="0">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr>
			<td>Gold series DuraShock Hand Aneroid Sphygmomanometer</td>
			<td>Welch Allyn</td>
			<td>DS56</td>
			<td>Manual Sphygmomanometer</td>
		</tr>
		<tr>
			<td>Neonate [size 2] 1 Tube, 10 Pack</td>
			<td>Welch Allyn</td>
			<td>5082-102-1&#160;</td>
			<td>Limb blood pressure cuff</td>
		</tr>
		<tr>
			<td>Luer lock adaptor</td>
			<td>Welch Allyn</td>
			<td>5082-178</td>
			<td>Adaptor for neonatal cuff</td>
		</tr>
		<tr>
			<td>Thermistor pod</td>
			<td>AD Instruments</td>
			<td>ML 309</td>
			<td>skin tempertature unit</td>
		</tr>
		<tr>
			<td>Skin temperture probe</td>
			<td>AD Instruments</td>
			<td>MLT 422/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Powerlab, 4 channel acquistion hardware</td>
			<td>AD Instruments</td>
			<td>PL 35044</td>
			<td>Skin temperature recorder</td>
		</tr>
		<tr>
			<td>Homeothermic blanket system with flexible probe</td>
			<td>Harvard Appartus</td>
			<td>507222F</td>
			<td></td>
		</tr>
		<tr>
			<td>Towel</td>
			<td></td>
			<td></td>
			<td>optional: awake remote ischemia</td>
		</tr>
		<tr>
			<td>Isoflo - 100% Isoflurane -250 ml</td>
			<td>Abbot Animal Health</td>
			<td>05260-05</td>
			<td>optional: inhaltion anaesthetic remote ischemia</td>
		</tr>
		<tr>
			<td>Ketamil - ketamine 100 mg/ml - 50 ml</td>
			<td>Troy Laboratories Pty Ltd</td>
			<td></td>
			<td>optional: injectable anaesthetic remote ischemia</td>
		</tr>
		<tr>
			<td>Xylium - Xylazine 100 mg/ml - 50 ml</td>
			<td>Troy Laboratories Pty Ltd</td>
			<td></td>
			<td>optional: injectable anaesthetic remote ischemia</td>
		</tr>
	</tbody>
</table>

remote limb ischemic preconditioning:  a neuroprotective technique in rodents

亚致死缺血保护免受随后，更严重的缺血组织通过内源性机制在受影响的组织的上调。亚致死缺血也已表明上调保护机制在远程组织。缺血（5-10分钟）在哺乳动物后肢的简短期间诱导的脑，肺，心脏和视网膜自我保护反应。该效应被称为远程缺血预处理（RIP）。它是保护重要器官的治疗有希望的途径，并且已经根据对心脏和脑损伤的临床试验。该出版物表明使肢体的控制，微创方法 - 大鼠的具体后肢 - 缺血。在人类新生儿开发用于一种血压箍被连接到一个手动血压计和用于施加围绕后肢的上部160毫米汞柱的压力。设计成检测皮肤温度的探针被用来验证ischemi一，通过记录下降引起的腿动脉压引起的闭塞皮肤温度和温度升高随后的压脉袋的释放。 RIP的这种方法提供保护，以对抗明亮光损伤和退行性变大鼠视网膜。

升高到高于160毫米汞柱的血压停止血流到后肢如在图1B中清楚地看到。组织氧合的缺乏导致了减少动物的脚温度为缺血再灌注协议（ 图2）。脚的温度（33℃）比核心温度低，可靠地减少在袖带压力升高（31℃）时，袖带放气（32℃）上升。单1000勒克斯的光损伤交付黯淡引发白鼠带或不带远程缺血性预处理。视网膜功能被记录并使用电图（ERG）进行评估。 <p class="jove_conte...

Watch this Scientific Journal Video about Remote Limb Ischemic Preconditioning:  A Neuroprotective Technique in Rodents at JoVE.com

Remote Limb Ischemic Preconditioning:  A Neuroprotective Technique in Rodents

远程缺血预处理（RIP）是调节组织反对破坏性压力的方法。我们已经建立了远程缺血的方法在后肢，由充气血压计袖带5-10分钟。 RIP的神经保护能力已经被证明在视网膜变性的啮齿类动物模型。

远程肢体缺血预适应：在啮齿动物的保护技术

Sublethal ischemia protects tissues against subsequent, more severe ischemia through the upregulation of endogenous mechanisms in the affected tissue. ...

remote-limb-ischemic-preconditioning-neuroprotective-technique

Research

JoVE Journal

Medicine

10.9K 次观看.  University of Sydney. 远程缺血预处理（RIP）是调节组织反对破坏性压力的方法。我们已经建立了远程缺血的方法在后肢，由充气血压计袖带5-10分钟。 RIP的神经保护能力已经被证明在视网膜变性的啮齿类动物模型。 

视频: Remote Limb Ischemic Preconditioning:  A Neuroprotective Technique in Rodents

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety.
Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential.
In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates.

A method of gene transfer for the treatment of ischemic heart failure is described using a swine model of myocardial infarction. Our simple and reproducible method enables us to readily evaluate the efficacy of various gene transfers with a very simple and reproducible way.

缺血性心力衰竭在临床前模型的转基因

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

科研

医学

A Simplified Technique for Producing an Ischemic Wound Model

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work well in non-ischemic wounds may not work in human diabetic wounds because vasculopathy is one major factor that hinders healing of these wounds. We published an article in 2007 describing a rabbit ear ischemic wound model created by a minimally invasive surgical technique. Since then, we have further simplified the procedure for easier operation. On one ear, three small skin incisions were made on the vascular pedicles, 1-2 cm from the ear base. The central artery was ligated and cut along with the nerve. The whole cranial bundle was cut and ligated, leaving only the caudal branch intact. A circumferential subcutaneous tunnel was made through the incisions, to cut subcutaneous tissues, muscles, nerves, and small vessels. The other ear was used as a non-ischemic control. Four wounds were made on the ventral side of each ear. This technique produces 4 ischemic wounds and 4 non-ischemic wounds in one animal for paired comparisons. After surgery, the ischemic ear was cool and cyanotic, and showed reduced movement and a lack of pulse in the ear artery. Skin temperature of the ischemic ear was 1-10 &deg;C lower than that on the normal ear and this difference was maintained for more than one month. Ear tissue high-energy phosphate contents were lower in the ischemic ear than the control ear. Wound healing times were longer in the ischemic ear than in the non-ischemic ear when the same treatment was used. The technique has now been used on more than 80 rabbits in which 23 were diabetic (diabetes time ranging from 2 weeks to 2 years). No single rabbit has developed any surgical complications such as bleeding, infection, or rupture in the skin incisions. The model has many advantages, such as little skin disruption, longer ischemic time, and higher success rate, when compared to many other models. It can be safely used in animals with reduced resistance, and can also be modified to meet different testing requirements.

We have developed a minimally invasive technique to create a rabbit ischemic ear wound model by dividing the central artery and nerve and the cranial neurovascular bundle. A subcutaneous tunnel then cuts all subcutaneous tissues. This procedure causes minimal skin disruption and can be safely used in diabetic animals.

一个生产缺血性损伤模型的简化技术

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work ...

Remote Magnetic Navigation for Accurate, Real-time Catheter Positioning and Ablation in Cardiac Electrophysiology Procedures

New remote navigation systems have been developed to improve current limitations of conventional manually guided catheter ablation in complex cardiac substrates such as left atrial flutter. This protocol describes all the clinical and invasive interventional steps performed during a human electrophysiological study and ablation to assess the accuracy, safety and real-time navigation of the Catheter Guidance, Control and Imaging (CGCI) system. Patients who underwent ablation of a right or left atrium flutter substrate were included. Specifically, data from three left atrial flutter and two counterclockwise right atrial flutter procedures are shown in this report. One representative left atrial flutter procedure is shown in the movie. This system is based on eight coil-core electromagnets, which generate a dynamic magnetic field focused on the heart. Remote navigation by rapid changes (msec) in the magnetic field magnitude and a very flexible magnetized catheter allow real-time closed-loop integration and accurate, stable positioning and ablation of the arrhythmogenic substrate.

This report provides a detailed description of a new remote navigation system based on magnetic driven forces, which has been recently introduced as a new robotic tool for human cardiac electrophysiology procedures.

遥控磁导航导管定位准确，实时和心脏电生理消融程序

New remote navigation systems have been developed to improve current limitations of conventional manually guided catheter ablation in complex cardiac ...

The Use of Cystometry in Small Rodents: A Study of Bladder Chemosensation

The lower urinary tract (LUT) functions as a dynamic reservoir that is able to store urine and to efficiently expel it at a convenient time. While storing urine, however, the bladder is exposed for prolonged periods to waste products. By acting as a tight barrier, the epithelial lining of the LUT, the urothelium, avoids re-absorption of harmful substances. Moreover, noxious chemicals stimulate the bladder's nociceptive innervation and initiate voiding contractions that expel the bladder's contents. Interestingly, the bladder's sensitivity to noxious chemicals has been used successfully in clinical practice, by intravesically infusing the TRPV1 agonist capsaicin to treat neurogenic bladder overactivity1. This underscores the advantage of viewing the bladder as a chemosensory organ and prompts for further clinical research. However, ethical issues severely limit the possibilities to perform, in human subjects, the invasive measurements that are necessary to unravel the molecular bases of LUT clinical pharmacology. A way to overcome this limitation is the use of several animal models2. Here we describe the implementation of cystometry in mice and rats, a technique that allows measuring the intravesical pressure in conditions of controlled bladder perfusion.
	After laparotomy, a catheter is implanted in the bladder dome and tunneled subcutaneously to the interscapular region. Then the bladder can be filled at a controlled rate, while the urethra is left free for micturition. During the repetitive cycles of filling and voiding, intravesical pressure can be measured via the implanted catheter. As such, the pressure changes can be quantified and analyzed. Moreover, simultaneous measurement of the voided volume allows distinguishing voiding contractions from non-voiding contractions3.
	Importantly, due to the differences in micturition control between rodents and humans, cystometric measurements in these animals have only limited translational value4. Nevertheless, they are quite instrumental in the study of bladder pathophysiology and pharmacology in experimental pre-clinical settings. Recent research using this technique has revealed the key role of novel molecular players in the mechano- and chemo-sensory properties of the bladder.

Cystometry is an efficient technique to measure bladder function of small animals in vivo. The bladder is continuously infused at rates controlled through an intravesical catheter, whereas the urethra is left free for micturition. This allows for repetitive filling and emptying of the bladder, while intravesical pressure and voided volume are recorded.

的的膀胱测压小老鼠：A膀胱Chemosensation研究的使用

	The lower urinary tract (LUT) functions as a  dynamic reservoir that is able to store urine and to efficiently expel it at a  convenient time. While ...

Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We have established bilateral common carotid artery occlusion as a preconditioning stimulus to induce early ischemic tolerance to transient focal cerebral ischemia in C57Bl6/J mice. In this video, we will demonstrate the methodology used for this study.

There is accumulating evidence, that ischemic preconditioning (PC) &#x2013; a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice.

双侧颈总动脉闭塞足够的预处理刺激诱导局灶性脑缺血早期缺血耐受

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a ...

Angiogenesis in the Ischemic Rat Lung

The adult lung is perfused by both the systemic bronchial artery and the entire venous return flowing through the pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that responds to a need for enhanced lung perfusion and shows robust neovascularization. Pulmonary vascular ischemia induced by pulmonary artery obstruction has been shown to result in rapid systemic arterial angiogenesis in man as well as in several animal models. Although the histologic assessment of the time course of bronchial artery proliferation in rats was carefully described by Weibel 1, mechanisms responsible for this organized growth of new vessels are not clear. We provide surgical details of inducing left pulmonary artery ischemia in the rat that leads to bronchial neovascularization. Quantification of the extent of angiogenesis presents an additional challenge due to the presence of the two vascular beds within the lung. Methods to determine functional angiogenesis based on labeled microsphere injections are provided.

The lung is perfused by both the systemic bronchial artery and pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that shows robust neovascularization. Cessation of pulmonary blood flow promotes brisk bronchial angiogenesis. We provide surgical details of inducing left pulmonary artery ischemia that promotes bronchial neovascularization.

在缺血大鼠肺的血管生成

The  adult lung is perfused by both the systemic bronchial artery and the entire  venous return flowing through the pulmonary arteries. In most lung ...

Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment epithelium (RPE) cells. RPE provide critical support for photoreceptors and death or dysfunction of RPE cells is characteristic of age-related macular degeneration (AMD), the leading cause of permanent vision loss in people age 55 and older. While no cure for AMD has been identified, implantation of healthy RPE in diseased eyes may prove to be an effective treatment, and large numbers of RPE cells can be readily generated from pluripotent stem cells. Several interesting questions regarding the safety and efficacy of RPE cell delivery can still be examined in animal models, and well-accepted protocols used to inject RPE have been developed. The technique described here has been used by multiple groups in various studies and involves first creating a hole in the eye with a sharp needle. Then a syringe with a blunt needle loaded with cells is inserted through the hole and passed through the vitreous until it gently touches the RPE. Using this injection method, which is relatively simple and requires minimal equipment, we achieve consistent and efficient integration of stem cell-derived RPE cells in between the host RPE that prevents significant amount of photoreceptor degeneration in animal models. While not part of the actual protocol, we also describe how to determine the extent of the trauma induced by the injection, and how to verify that the cells were injected into the subretinal space using in vivo imaging modalities. Finally, the use of this protocol is not limited to RPE cells; it may be used to inject any compound or cell into the subretinal space.

Here we present a community accepted protocol in multimedia format for subretinally injecting a bolus of RPE cells in rats and mice. This approach can be used for determining rescue potentials, safety profiles, and survival capacities of grafted RPE cells upon implantation in animal models of retinal degeneration.

视网膜下进行注射的鼠害提供视网膜色素上皮细胞悬浮

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment ...

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

新生儿缺氧缺血性脑病的小猪模型

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

A Method for Quantifying Upper Limb Performance in Daily Life Using Accelerometers

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. It has become important to measure a person's activities in daily life, and not just measure their capacity for activity in the structured environment of a clinic or laboratory. A wearable sensor that is now enabling measurement of daily movement is the accelerometer. Accelerometers are commercially-available devices resembling large wrist watches that can be worn throughout the day. Data from accelerometers can quantify how the limbs are engaged to perform activities in peoples' homes and communities. This report describes a methodology to collect accelerometry data and turn it into clinically-relevant information. First, data are collected by having the participant wear two accelerometers (one on each wrist) for 24 h or longer. The accelerometry data are then downloaded and processed to produce four different variables that describe key aspects of upper limb activity in daily life: hours of use, use ratio, magnitude ratio, and the bilateral magnitude. Density plots can be constructed that visually represent the data from the 24 h wearing period. The variables and their resultant density plots are highly consistent in neurologically-intact, community-dwelling adults. This striking consistency makes them a useful tool for determining if upper limb daily performance is different from normal. This methodology is appropriate for research studies investigating upper limb dysfunction and interventions designed to improve upper limb performance in daily life in people with stroke and other patient populations. Because of its relative simplicity, it may not be long before it is also incorporated in clinical neurorehabilitation practice.

This protocol describes a method to quantify upper limb performance in daily life using wrist-worn accelerometers.

一种使用加速度计量化上肢表现在日常生活的方法

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. ...

Remote Laboratory Management: Respiratory Virus Diagnostics

An uptick in recent pandemics (Ebola, Zika, MERS, influenza, etc.) underlines the need for a more 'nimble,' coordinated response that addresses a multitude of issues ranging from transportation, access, facilities, equipment, and communication to provider training. To address this need, we have developed an innovative, scalable, logistics-enhanced, mobile, laboratory facility for emergencies and epidemics in resource-constrained global settings. Utilizing a background in clinical operations as an academic medical center, we designed a rapidly-deployable, modular BSL-2 and BSL-3 facility with user-friendly software for tracking and management of drugs and supplies in remote regions during epidemics and outbreaks. Here, we present our intermodal, mobile, expandable shipping-container laboratory units. The design of the laboratory facilitates off-grid usage by minimizing power consumption and allowing alternate water sources. The unit's information communication technology (ICT) platform provides (i) user-friendly tablet-based documentation, (ii) enhanced tracking of patients and supplies, and (iii) integrated communication onsite with built-in telehealth capabilities. To ensure quality in remote environments, we have developed a checklist for a basic laboratory workflow and a protocol for respiratory viral diagnosis using reverse-transcription polymerase chain reaction (RT-PCR). As described, this innovative and comprehensive approach allows for the provision of laboratory capability in resource-limited global environments.

A rapidly-deployable, off-grid laboratory has been designed and built for remote, resource-constrained global settings. The features and critical aspects of the logistically-enhanced, expandable, multifunctional laboratory modules are explored. A checklist for a basic laboratory workflow and a protocol for a respiratory viral diagnostic test are developed and presented.