这项工作是由美国国立卫生研究院（NIH）K12的研究院物理及职业治疗师综合康复研究训练机会（K12的HD055931），物理疗法和匹兹堡克劳德D.辣椒老年人美国人独立中心基金会（P29 AG024827 ）。

1。电极的制备<ul><li>切载体电路板1厘米×1厘米一节。 </li><li>稳定2 wirewrap引脚副把手，使用间隔约3.5毫米外销。引脚岔完应朝上。 </li><li>弯曲在90°角wirewrap针，约一半沿其长度，使用needlenose钳子。 </li><li>揭露连接器，导线连接约7厘米，剥电线绝缘的铜线。 </li><li>锡铜线之一wirewrap引脚岔。重复为其他wirewrap引脚焊接一步。 </li><li>拉收缩铜线和金销之间的焊接连接管，以隔离和稳定。使用烙铁头加热管。 </li><li>插入相邻线路板电路板开口的wirewrap引脚unsoldered提示。 </li><li>确保了wirewrap引脚是相互平行和STICK提示位于相同的距离，通过放置在电路板时，通过线路板等。 </li><li>嵌入在清晰环氧树脂的引脚焊完。允许环氧树脂固化约10分钟，或直至完全干燥。 </li><li>重复直到载体的引脚和电路板，以提供结构完整性和导线应变消除完全嵌入环氧树脂应用。 </li><li>沙下环氧树脂使用一个金属文件揭露黄金引脚的提示。 </li><li>用万用表的两根导线，以确保电气短路和导线有适当的连续性</li><li>将电极从神经肌肉电刺激装置，导致女性2路电线连接器（波莫纳跳线）。 </li></ul> 2。动物的制备<ul><li>动物是通过2的％isofluorane（雅培，北芝加哥，IL）使用100％的O 2气体吸入麻醉。动物应保持整个ţ麻醉他NMES会议。 </li><li>在开始之前，神经肌肉电刺激协议执行脚趾捏，以确保该动物是完全麻醉。 </li><li>动物应定期检查，以确保麻醉的水平是足够的刺激。麻醉应根据需要进行调整。运动刺激的腿，呼吸深度的变化，和粘膜的颜色都应该定期评估监测麻醉深度。 </li><li>放置侧卧动物。 </li><li>适用于眼科润滑剂在手术过程中，以防止角膜干燥的眼睛。 </li><li>刮胡子后肢区域来对待，用酒精擦拭。 </li><li>用3％漂白粉擦拭电极，然后用清水冲洗干净。 </li><li>应用层进行了电极将应用于网站的凝胶。必要再涂整个神经肌肉电刺激协议，以确保电极与皮肤之间的电流流动。 </li><li>一个热灯Øŕ应使用循环水毯，以维持正常的身体核心范围。 </li><li> 注：由美国匹兹堡大学制度的动物护理和使用委员会已审查和批准的所有程序，在保证小灵通和AAALAC认证诠释演出。认可的程序和设施。 </li></ul> 3。刺激<ul><li>电极的位置： <ul><li>前车厢的肌肉，包括在胫骨前和，伸趾长肌的肌肉刺激，将直接通过动物的深腓骨神经，1远端分支腓总神经断的表面电极，是位于只是前了腓骨头。 </li><li>前室肌肉的刺激，放置电极确认时刺激引起全踝关节背屈和数字扩展。另一方面，背屈位分机的情况下，建议只胫事前rior肌肉受到刺激。这种有针对性的收缩反应，可能是可取的，这取决于研究设计。 </li><li>肌肉收缩分为2个阶段的刺激：一个自由的移动，同心阶段，在最初的发生〜0.5秒。在第一阶段，爪子移动到最大背屈位分机从静止位置。第二阶段的刺激是在结束范围背屈位分机持续等长收缩。 </li></ul></li><li>刺激参数： <ul><li>神经肌肉电刺激模式采用300光伏EMPI多功能电疗设备，它提供了传统的神经肌肉电刺激2通道（见表）。对于这种模式的目的，只有1个通道。 </li><li>使用的参数包括对称波形，150μs的脉冲持续时间和频率为50Hz。刺激的时间设置为5秒，0.5秒的上升和0.5秒的斜坡下降。这将使肌肉逐渐accli队友来刺激。在目前的协议，关闭之间的收缩时间设置为10秒，但是这取决于预期的效果可能会有所调整。减少关闭时间将导致更快速的启动了肌肉疲劳。这些刺激参数，根据临床方案设计，以提高使用神经肌肉电刺激肌肉力量，不引起显着的骨骼肌损伤1，15，16。 </li><li>小鼠完成两套10收缩，在5分钟的休息间套。 </li><li>对于成年动物，神经肌肉电刺激强度通常始于9毫安。从我们的经验，这是接近最大强度不引起明显的步态障碍，紧随刺激。为会议后续神经肌肉电刺激，强度增加1 mA的动物是能够完成20个完整的dorsiflexions每次。 </li></ul></li><li>刺激协议和RECO完成后非常麻醉，动物通常表现出正常的步态和姿势。步态应该继续受到损害整个神经肌肉电刺激计划的持续时间。 </li></ul> 4。代表结果已实施几个小鼠品系，包括：野生型（B6/10），B6.SCID和MDX / SCID小鼠神经肌肉电刺激在这篇文章中描述的协议。 MDX / SCID的老代表进行了4周（20届）在3-5个月的神经肌肉电刺激结果提出。 动物人道安乐死，而麻醉下颈椎脱位。胫骨前肌收获，并立即冻结在2甲基丁烷在液氮预冷，并保存在-80°C。串行的横截面（10微米），获得并安装到幻灯片。使用标准的统计软件包（SPSS软件，v19.0软件）进行统计分析。首先，列文的测试是用来评估是否有WA差异的平等。独立样本t检验，然后进行探讨神经肌肉电刺激组和对照组之间的差异。 苏木精伊红（H＆E）的污渍进行了探讨神经肌肉电刺激的协议是否会导致营养不良肌肉损伤增加。选择一节，并获得了使用光学显微镜（日本）;尼康，尼康的Eclipse E800的图片。手工计算，使用国立卫生研究院（NIH） - 开发的图像分析软件，图像J.再生指数没有显着增加（中央核纤维纤维和位于市中心的核纤维数量总数/总数在提交给神经肌肉电刺激动物纤维），较对照组（P = 0.802; 图1）。这表明，神经肌肉电刺激的应用不会增加在营养不良的动物中观察到的变性再生级联，因此不可能被诱导增加​​肌肉损伤。 CD31的免疫染色进行探讨肌肉血管的4周神经肌肉电刺激协议的影响。简言之，肌肉部分被固定在4％福尔马林和阻止使用5％马血清（HS）目录。部分培养与鼠抗鼠抗体（1:300稀释于5％HS）和555标记的羊抗鼠二抗（1:300稀释于5％HS）治疗。量化的CD31阳性细胞总数，一节被选中，并拍下使用荧光显微镜（日本尼康的Eclipse E800的）。毛细血管的总人数是手动计算使用北Eclipse软件（Empix影像公司）。有一个在提交给神经肌肉电刺激动物的CD31阳性细胞数量显着增加，比对照组（P &lt;0.01，图2）时，表明，神经肌肉电刺激协议描述促进骨骼肌血管生成。 <stro吴&gt; 在原位收缩测试：四个星期的神经肌肉电刺激协议完成后，车厢前部肌肉收缩测试使用原位测试仪器（型号809B，极光科技公司，加拿大），刺激（型号701C，极光科技有限公司，加拿大），力传感器（极光科技公司，加拿大）。简言之，通过一个小切口膝外侧腓总神经的麻醉动物分离。小鼠仰卧，然后放置在一个平台上，脚被测试踏板的位置上。膝盖和脚用布带用于测试后肢稳定。使用钩插入皮下的电极刺激肌肉。肌肉强直收缩，在150Hz频率为350MS测试调查4周神经肌肉电刺激的协议是否会诱发肌肉力量改善。结果正常化湿肌肉的重量。约30％的增幅在tetanIC提交给神经肌肉电刺激动物的收缩时，比对照组（P = 0.005）（ 图3），表明所述的协议，提高了MDX / SCID的动物骨骼肌肉力量。 <img alt="图1" src="/files/ftp_upload/3914/3914fig1.jpg" /> 图1苏木精和曙红营养不良骨骼肌染色（MDX）/免疫缺陷小鼠（4-5月）（10倍放大）：一）未经处理的控制，二）经过4周的神经肌肉电刺激。 <img alt="图2" src="/files/ftp_upload/3914/3914fig2.jpg" /> 图2。4周后，CD31的免疫（红色），作为骨骼肌血管标记，在营养不良的骨骼肌（MDX）/免疫缺陷小鼠（4-5月）（20倍放大）一）未经处理的控制，二）神经肌肉电刺激。 <img alt="图3" src="/files/ftp_upload/3914/3914fig3.jpg" /> 收缩测试图3。在控制前车厢肌肉和4周神经肌肉电刺激治疗的动物。 MNM =毫牛顿米。 <a href="http://www.jove.com/files/ftp_upload/3914/3914fig3large.jpg">点击这里查看大图</a> 。

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我们什么都没有透露。

这些结果表明，我们发达的小鼠模型的神经肌肉电刺激促进骨骼肌血管生成（ 图2）和加强（ 图3），但不引起骨骼肌肉损伤（ 图1）。 应当指出的是，这里所描述的刺激参数已超载车厢前部肌肉引起肌肉。就像是为临床情景的情况下，电极位置可能会有所调整，以刺激其他肌肉群，虽然肌肉到神经肌肉电刺激的反应可能会有所不同，取决于纤维类型组成。根据研究设计，神经肌肉电刺激的持续时间，疗程的总人数，以及重复的总数可能会修改。 这种方法有任何协议，应注意限制。在本研究中，进行深腓骨神经刺激。然而，在诊所，刺激我小号通常在电机单位管理。在动物模型，对神经的刺激将需要较低的强度，以引起全肌肉收缩。作为提出的模型的另一个限制是，我们无法获得有关的舒适程度，在神经肌肉电刺激的应用，给动物麻醉。因此，在临床人口可比强度的耐受性是难以估量。然而，我们的组织学研究结果表明我们的神经肌肉电刺激模型，描述的那样，不会引起肌肉损伤。 动物模型，模拟通常在门诊实施的方式提供有用的实验工具，可以更好地了解细胞和分子反应治疗干预的发展。此外，这种模式是有用的，要进行临床前研究，同时完善现有的康复协议，并开发新的迹象。

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>矢量线岔焊接端子的包装职位</td><td>纽瓦克</td><td> T68 </td></tr><tr><td>矢量电路板</td><td>纽瓦克</td><td> 3662-2 </td></tr><tr><td>波诺马修补线</td><td>纽瓦克</td><td>的P-36-0 </td></tr><tr><td> 5分钟的环氧树脂</td><td>旁白贝克公司</td><td> 4001 </td></tr><tr><td>光谱360电极凝胶</td><td>米利肯医学</td><td> MR41217 </td></tr><tr><td>便携式神经肌肉刺激装置</td><td> EMPI </td><td> 300pv </td></tr></tbody></table>

a murine model of muscle training by neuromuscular electrical stimulation

神经肌肉电刺激（NMES）是临床上常见的方式，被广泛用于恢复，保持或提高3-5肌功能的能力。皮的表面刺激骨骼肌涉及阴极和阳极之间的电流流动，从而引起电机的单位和周围的肌肉纤维的兴奋。 神经肌肉电刺激是一个有吸引力的方式来评估骨骼肌的适应性反应，有以下几个原因。首先，它提供了一个可重复的实验模型，是有据可查的生理适应，如肌纤维hypertophy和6加强肌肉，血管生成7-9，9-11生长因子的分泌和肌肉前体细胞活化12。这种生理反应，可仔细滴定使用不同的刺激参数（Cochrane回顾，见13）。此外，NMES新兵运动单位的非选择性，在一个固定的空间和时间同步的方式14，提供所有纤维的治疗效果发挥优势，无论纤维类型。虽然也有特定的禁忌症，在临床的人口，包括周边静脉疾病或恶性肿瘤，神经肌肉电刺激，例如，神经肌肉电刺激是安全和可行的，甚至为那些生病和/或长期卧床和严格锻炼可具有挑战性的人群。 在这里，我们展示市售电极适应和利用小鼠模型进行神经肌肉电刺激协议的协议。这种动物模型利用临床上​​可用的设备，并提供即时反馈电极引出所需的肌肉收缩作用的定位优势。本手稿的目的，我们将描述为鼠标后肢前车厢肌肉刺激肌肉的协议。

Watch this Scientific Journal Video about A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation at JoVE.com

A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

小鼠模型的神经肌肉电刺激（NMES），临床安全和廉价的方式，车厢前部肌肉被描述。这种模式有针对性和具体的肌肉收缩引发小鼠的目的修改一个现成的临床设备的优势。

一个小鼠模型，通过神经肌肉电刺激肌肉训练

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional ...

a-murine-model-muscle-training-neuromuscular-electrical

Research

JoVE Journal

Medicine

20.8K 次观看.  University of Pittsburgh. 小鼠模型的神经肌肉电刺激（NMES），临床安全和廉价的方式，车厢前部肌肉被描述。这种模式有针对性和具体的肌肉收缩引发小鼠的目的修改一个现成的临床设备的优势。

视频: A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

Programmed Electrical Stimulation in Mice

Genetically-modified mice have emerged as a preferable animal model to study the molecular mechanisms underlying conduction abnormalities, atrial and ventricular arrhythmias, and sudden cardiac death.1 Intracardiac pacing studies can be performed in mice using a 1.1F octapolar catheter inserted into the jugular vein, and advanced into the right atrium and ventricle. Here, we illustrate the steps involved in performing programmed electrical stimulation in mice. Surface ECG and intracardiac electrograms are recorded simultaneously in the atria, atrioventricular junction, and ventricular myocardium, whereas intracardiac pacing of the atrium is performed using an external stimulator. Thus, programmed electrical stimulation in mice provides unique opportunities to explore molecular mechanisms underlying conduction defects and cardiac arrhythmias.

Programmed electrical stimulation provides the ability to determine conduction properties of the heart, and the possibility to induce and terminate cardiac arrhythmias using various pacing protocols. Using a transvenous catheter, intracardiac electrogram recordings can be obtained in mice following programmed electrical stimulation protocols to identify arrhythmogenic substrates.

程控电刺激小鼠

Genetically-modified mice have emerged as a preferable animal model to study the molecular mechanisms underlying conduction abnormalities, atrial and ...

科研

医学

Utilizing Transcranial Magnetic Stimulation to Study the Human Neuromuscular System

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade. While the use of TMS has expanded to the study of many systems and processes during this time, the original application and perhaps one of the most common uses of TMS involves studying the physiology, plasticity and function of the human neuromuscular system. Single pulse TMS applied to the motor cortex excites pyramidal neurons transsynaptically 2 (Figure 1) and results in a measurable electromyographic response that can be used to study and evaluate the integrity and excitability of the corticospinal tract in humans 3. Additionally, recent advances in magnetic stimulation now allows for partitioning of cortical versus spinal excitability 4,5. For example, paired-pulse TMS can be used to assess intracortical facilitatory and inhibitory properties by combining a conditioning stimulus and a test stimulus at different interstimulus intervals 3,4,6-8. In this video article we will demonstrate the methodological and technical aspects of these techniques. Specifically, we will demonstrate single-pulse and paired-pulse TMS techniques as applied to the flexor carpi radialis (FCR) muscle as well as the erector spinae (ES) musculature. Our laboratory studies the FCR muscle as it is of interest to our research on the effects of wrist-hand cast immobilization on reduced muscle performance6,9, and we study the ES muscles due to these muscles clinical relevance as it relates to low back pain8. With this stated, we should note that TMS has been used to study many muscles of the hand, arm and legs, and should iterate that our demonstrations in the FCR and ES muscle groups are only selected examples of TMS being used to study the human neuromuscular system.

Transcranial magnetic stimulation (TMS) is a non-invasive tool to gain insight on the physiology and function of the human nervous system. Here, we present our TMS techniques to study cortical excitability of the upper limb and lumbar musculature.

利用经颅磁刺激，以研究人类的神经肌肉系统

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade.  While ...

Breathing-controlled Electrical Stimulation (BreEStim) for Management of Neuropathic Pain and Spasticity

Electrical stimulation (EStim) refers to the application of electrical current to muscles or nerves in order to achieve functional and therapeutic goals. It has been extensively used in various clinical settings. Based upon recent discoveries related to the systemic effects of voluntary breathing and intrinsic physiological interactions among systems during voluntary breathing, a new EStim protocol, Breathing-controlled Electrical Stimulation (BreEStim), has been developed to augment the effects of electrical stimulation. In BreEStim, a single-pulse electrical stimulus is triggered and delivered to the target area when the airflow rate of an isolated voluntary inspiration reaches the threshold. BreEStim integrates intrinsic physiological interactions that are activated during voluntary breathing and has demonstrated excellent clinical efficacy. Two representative applications of BreEStim are reported with detailed protocols: management of post-stroke finger flexor spasticity and neuropathic pain in spinal cord injury.

Breathing-controlled Electrical Stimulation BreEStim for Management of Neuropathic Pain and Spasticity

The purpose is to present a new method, breathing-control electrical stimulation (BreEStim) for management of neuropathic pain and spasticity.

呼吸控制电气的刺激（BreEStim）的神经性疼痛和痉挛的管理

Electrical stimulation (EStim) refers to the application of electrical current to muscles or nerves in order to achieve functional and therapeutic goals. ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

伤口愈合的小鼠模型

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

蛛网膜下腔出血的小鼠模型

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Background 
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered advancements to decrease the occurrence this injury.&#160;The current model demonstrates reproducible lower&#160;extremity paralysis following thoracic aortic occlusion.
Methods 
Adult male C57BL6 mice were anesthetized with isoflurane. Through a cervicosternal incision the aorta was exposed. The descending thoracic aorta and left subclavian arteries were identified without entrance into pleural space. Skeletonization of these arteries was followed by immediate closure (Sham) or occlusion for 4 min (moderate ischemia) or 8 min (prolonged ischemia). The sternotomy and skin were closed and the mouse was transferred to warming bed for recovery.&#160; Following recovery, functional analysis was obtained at 12 hr intervals until 48 hr.
Results 
Mice that underwent sham surgery showed no observable hind&#160;limb deficit. Mice subjected to moderate ischemia for 4 min had minimal functional deficit at 12 hr followed by progression to complete paralysis at 48 hr. Mice subjected to prolonged ischemia had an immediate paralysis with no observable hind-limb movement at any point in the postoperative period. There was no observed intraoperative or post&#160;operative mortality.
Conclusion 
Reproducible lower extremity paralysis whether immediate or delayed can be achieved in a murine model. Additionally, by using a median sternotomy and careful dissection, high survival rates, and reproducibility can be achieved.

The lack of mechanistic understanding of spinal cord ischemia-reperfusion injury has hindered further adjuncts to prevent paraplegia following high risk aortic operations. Thus, the development of animal models is imperative. This manuscript demonstrates reproducible lower extremity paralysis following thoracic aortic occlusion in a murine model.

可重放截瘫胸主动脉阻断脊髓缺血再灌注的小鼠模型

Background
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered ...

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.
A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.
Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic&#160;and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.
Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

右心室结构和功能的肺动脉缩窄经胸超声小鼠模型的评估

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As such, primary or &#39;neuronal-like&#39; cell culture systems are commonly utilized. While&#160;these systems are relatively easy to work with, and are useful model systems in which various functional outcomes (such as cell death) can be readily quantified, the examined outcomes and pathways in cultured immature neurons (such as excitotoxicity-mediated cell death pathways) are not necessarily the same as those observed in mature brain, or in intact tissue. Therefore, there is the need to develop models in which cellular mechanisms in mature neural tissue can be examined. We have developed an in vitro technique that can be used to investigate a variety of molecular pathways in intact nervous tissue. The technique described herein utilizes rat cortical tissue, but this technique can be adapted to use tissue from a variety of species (such as mouse, rabbit, guinea pig, and chicken) or brain regions (for example, hippocampus, striatum, etc.). Additionally, a variety of stimulations/treatments can be used (for example, excitotoxic, administration of inhibitors, etc.). In conclusion, the brain slice model described herein can be used to examine a variety of molecular mechanisms involved in excitotoxicity-mediated brain injury.

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

We have developed a brain slice model which can be used to examine molecular mechanisms involved in excitotoxicity-mediated brain injury.&#160;This technique generates viable mature brain tissue and reduces animal numbers required for experimentation, whilst keeping the neuronal circuitry, cellular interactions, and postsynaptic compartments partly intact.

脑Microslices的兴奋性刺激作为<em&gt;在体外</em&gt;中风模型

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

Clinical Assessment of Spatiotemporal Gait Parameters in Patients and Older Adults

Spatial and temporal characteristics of human walking are frequently evaluated to identify possible gait impairments, mainly in orthopedic and neurological patients1-4, but also in healthy older adults5,6. The quantitative gait analysis described in this protocol is performed with a recently-introduced photoelectric system (see&#160;Materials table) which has the potential to be used in the clinic because it is portable, easy to set up (no subject preparation is required before a test), and does not require maintenance and sensor calibration. The photoelectric system consists of series of high-density floor-based photoelectric cells with light-emitting and light-receiving diodes that are placed parallel to each other to create a corridor, and are oriented perpendicular to the line of progression7. The system simply detects interruptions in light signal, for instance due to the presence of feet within the recording area. Temporal gait parameters and 1D spatial coordinates of consecutive steps are subsequently calculated to provide common gait parameters such as step length, single limb support and walking velocity8, whose validity against a criterion instrument has recently been demonstrated7,9. The measurement procedures are very straightforward; a single patient can be tested in less than 5 min&#160;and a comprehensive report can be generated in less than 1&#160;min.

This protocol is used to evaluate spatial and temporal gait variables of neurological/orthopedic patients and older persons by means of a recently-introduced floor-based photocell system.

时空步态参数的患者和老年人的临床评估

Spatial and temporal characteristics of human walking are frequently evaluated to identify possible gait impairments, mainly in orthopedic and ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.