这项工作是由骨科遗产基金会公元前克拉克的资助部分经费。我们希望国家​​特别感谢她的协助您玛丽莎麦克金尼创造许多数字图形。

1。 FCR和ES肌肉的单和双脉冲TMS <ol><li> 基本安全预防措施：在此之前执行TMS在一个人的主体性是必要的，因为它涉及到基本的安全预防措施，他们的第一个屏幕接触磁场。在我们的实验室中，我们遵循筛选磁共振安全， 教育和研究10研究所提出的指导方针。在我们的实验室中，我们也经常与癫痫发作的家族病史的人排除在外。我们还要求受试者接受TMS的ES肌肉戴上耳塞和一张嘴后卫的重点和更有力的刺激强度。 </li><li> 电气录音：要检查是必要的记录骨骼肌肌电图（EMG）信号在电机系统TMS的反应。对于FCR肌表面电极放在前臂上使用双极电极在T纵向安排位于他的肌肉光头和磨损皮肤，正如我们先前所描述的7,11。对于竖脊肌的肌肉，我们使用类似的电极位于剃光和磨损皮肤8在L3 - L5椎体水平的肌肉纵向安排。 </li><li> TMS线圈方向：主要激活皮质的神经元，transsynaptically它是必要的适当位置的TMS线圈12。对于FCR的肌肉，我们把一个70毫米的数字“8 TMS线圈相切的头皮和中线45度，所以，在侧后方内侧前方向的感应电流的流动。对于ES肌肉中，我们使用一个双锥形线圈，具有更大的穿透深度，是由于这些肌肉homunculus更深的代表性需要。在这里，线圈定位如在眼前的电流流入后的方向。我们有自定义的修改与激光附件系统我们的线圈，以协助我们在Subsequent重新定位的双锥形线圈。 </li><li> 识别“热点”：这是必要的，以确定引出最大的运动诱发电位的刺激位置。 FCR的肌肉，我们通过巧妙的移动TMS的线圈周围非常小的增量，并确定在我们观察到的最大的运动诱发电位振幅。一旦找到，我们注意到了不可磨灭的油墨，在头皮或莱卡第这方面。 TMS的ES肌肉TMS上肢肌肉比人类受试者相当多的不舒服。因此，我们简化了我们的ES肌肉的TMS的协议，以增加它的耐受性和可行性。在这里，而不是寻找“热点”我们用人体测量，以确定顶点的头骨。具体来说，我们确定了在矢状交集的头骨顶点（nasinon和枕外隆凸尖之间）和日冕（耳屏之间）飞机。 </li><li> 生物力学的定位： </在我们的实验室STRONG&gt;当我们执行TMS FCR的肌肉科目的手臂搁在一个BioDex系统4测功机（图2）扩展位置坐下。但是，我们应该注意，这只是一个可能的设置测量施加力量的例子。为ES肌肉科目要求一个直立的姿势坐下来与他们的手在他们的腿上休息（图3）。他们坐在旋转底座椅子上，大腿90 °相对躯干，小腿〜45 °相对的大腿上，在一个中立的posture8腰椎。 </li><li> 量化运动阈值 ：FCR的，我们提供单脉冲，在逐步增加刺激强度，直至运动诱发电位峰-峰值幅度超过50％的试验的大于50毫伏（图4）确定电机阈值（ MT） 。为了简化的TMS协议和提高耐受性和可行性，我们不运动阈值确定在E与相同的精度，当我们测试上肢肌肉的肌肉。相反，我们开始由50％的最大刺激输出提供一个初始的单脉冲，以确定是否这个刺激的强度是高于或低于运动阈值的TMS的协议。如果MEP是在观察到这种刺激强度的背景水平相对可辨别的MEP定义肌电图强度降低，以刺激输出的40％，以确定是否这个刺激的强度是子或超阈值 8 。 </li><li>我们提供一个单脉冲TMS的强度等于130％运动阈值的“热点”的FCR 量化MEP波幅使用单脉冲TMS：检查电机诱发电位波幅，并计算峰-峰幅度。一般情况下，我们正常化的最大复合肌肉纤维的动作电位超最大的正中神经电刺激后，这一结果。我们应该注意到，MEP的大小是版Ÿ皮质兴奋程度的依赖。因此，当TMS脉冲的背景下收缩期间交付，皮质兴奋性增加时，环境保护部大小将显着增加。对于ES肌肉，我们提供了一个单TMS脉冲强度为40或50％以上子运动阈值烈度8顶点。不幸的是，因为末梢神经支配ES肌肉电刺激不能访问，我们不能够正常化，这些运动诱发电位的复合型肌纤维的动作电位。 </li><li> 量化缄默期持续时间，采用单脉冲 TMS的：当一个TMS脉冲皮层交付过程中肌肉收缩，它会产生一个电气静止恢复活动前即指示皮质抑制，通常被称为沉默的运动诱发电位13期（图5）。要量化的缄默期，我​​们提供一个单一TMS脉冲“热点”等于130％运动阈值的强度，同时研究参与者是其最大强度的15％执行一个屈腕肌肉收缩。以前我们没有量化的ES肌肉的缄默期时间，但是，我们应该注意到，我们有传言称观察它的存在，在这个肌肉群时TMS脉冲ID的背景下收缩期间交付。 </li><li> 量化Intracortical便利使用双脉冲TMS的：我们使用双脉冲TMS量化intracortical便利6,7（图6和7的FCR和ES的肌肉，分别代表本测量）。 FCR的肌肉，我们首先要确定需要引出一个运动诱发电位之间的0.5-1.0 mV的刺激强度。下一步，我们提供了一个亚阈值调节脉冲在我们的实验室通常设置为等于70％运动阈值15毫秒阈上测试脉冲前。这一调理在这段时间内的测试脉冲前交付的脉冲将增加，或促进，电机的振幅诱发电位无条件相同的强度比单一脉冲。为ES肌肉群的调节脉冲强度设置为观察到的子运动阈值的强度（40％或50％的刺激输出），并设置测试脉冲强度分运动阈值水平（80％至40％以上或刺激产出的90％）8 。我们应该注意到，调节脉冲的强度可以根据这项研究的目的是多种多样的。同样，脉冲间隔可以改变肌肉和其相对位置的皮层而定。 </li><li> 量化短间隔Intracortical抑制使用双脉冲TMS的：我们也使用双脉冲TMS的量化短间隔intracortical抑制6,7（如图6日和7代表的FCR和ES肌肉的测量，分别） 。在这里，无论是FCR和ES的肌肉，程序是相同的测量两个脉冲之间的刺激间隔减少到3毫秒的异常intracortical便利的描述。在这段时间之前的测试脉冲发表这空调脉冲将减少，或抑制，电机的振幅诱发电位无条件相同的强度比单一脉冲。 </li><li> 量化长区间Intracortical抑制使用双脉冲TMS：提供两个相同的阈上测试是由100毫秒分开的脉冲也可以用来评估长的时间间隔intracortical 抑制 6,7 。在这种情况下，FCR的肌肉，第二个脉冲的运动诱发电位较小，或抑制更比第一（图8）相关，。我们以前没有量化intracortical抑制长的时间间隔在ES肌肉因超过受耐受关注。 </li></ol> 2。代表性的成果： 以下提供一个阈上TMS脉冲，刺激肌肉应该表现出一个很容易观察到的肌电响应（MEP）的（图4-8所示）。尽管它应该刺激发作和环境保护部之间的延迟正在审议的肌肉群之间会有所不同，但FCR的一般是16-19毫秒（图6）和ES的是17-22毫秒（图7;应当指出，明确环境保护部在ES肌肉开始在一些学科更难以直观地识别）。应该指出，当测试的ES肌肉群的其他几个肌肉群也明显随之而来，极大地刺激（包括下肢homunculus一般在同一地区，这是代表肌肉）。在测量intracortical便利MEP波幅通常比单一无条件脉冲（F观察igure 6和7​​）。然而，这是我们的经验，肌肉群之间的变化与一些肌肉群 - 如FCR显示在很多科目只有轻微的便利程度的便利。对于短间隔和长间隔intracortical抑制测量MEP波幅下降一般是观察比较单一相同的强度（图6-8）无条件脉冲。 <img alt="图1" src="/files/ftp_upload/3387/3387fig1.jpg" /> 图1。TMS的基本机制。TMS线圈感应磁场，穿透头皮和运动皮层内引起涡流。此涡流能够刺激大脑内的神经元。图转载自麦克金尼和克拉克，按14。 <img alt="图2" src="/files/ftp_upload/3387/3387fig2.jpg" /> 图2 T执行安装。<html> MS FCR的肌。请注意记录肌电（EMG）从前臂的信号，并在运动皮层的TMS的桨。我们一般也创纪录的肌肉力量，并使用电气周围神经刺激，以获得最大的复合肌纤维动作电位，因为这是在解释振幅值（例如，一个可以表达和MEP相对最大肌肉反应，而不是一个绝对有用mV值可大量皮下脂肪组织，如非生理因素）的影响。从下面的图转载：Clark等。 2008年9月 ，克拉克等，2010年6月 ，和麦克金尼等。 2010年7。 <img alt="图3" src="/files/ftp_upload/3387/3387fig3.jpg" /> 图3。执行TMS上竖spinale肌肉的设置 。戈斯等重印图。 2011 8。 _upload/3387/3387fig4.jpg“/&gt; 图4的运动阈值测定的例子。肌电图的痕迹代表诱发电位（MEP）的反应逐渐增加刺激强度（作为刺激输出（SO）的百分比表示）的电机。请注意，在低强度（28-30％的SO）非常小的MEP引起（亚阈值），但在32％，所以一个MEP引起电机，达到阈值（通常作为一个PP幅度MEP定义&gt; 50μV）。 图转载自麦克金尼和克拉克，按14。 <img alt="图5" src="/files/ftp_upload/3387/3387fig5.jpg" /> 图5的TMS：在收缩运动诱发潜在的缄默期。当对象执行一个单一的刺激，适用于运动皮层的轻微收缩和缄默期。第一部分是缄默期由于脊髓抑制，而后者则部分是由于大脑皮层的抑制，特别是GABA B受体。有没有达成共识的方式来量化的缄默期的持续时间，但我们的研究结果表明，无论是从刺激发作或MEP发病自愿的干扰肌电信号返回定义它是最可靠的的15。 图从2011年16克拉克和快速和麦克金尼和克拉克，按14重印。 <img alt="图6" src="/files/ftp_upload/3387/3387fig6.jpg" /> 图6：在电机的变化引起的潜力大小的FCR肌肉的第i个成对的脉冲TMS的。测量短的时间间隔intracortical抑制（SICI）和intracortical便利（ICF）。要量化SICI和ICF的调节脉冲（CP）运动阈值以下，并测试脉冲（TP）的设置，以唤起人们之间的0.5-1 mV的MEP的。在很短的interstimulus间隔（例如，3毫秒）CP抑制比较只到TP（SICI）环境保护部，而在较长的interstimulus间隔（如15毫秒），它促进了环境保护部“（ICF）。 CP：调节脉冲，总磷：测试脉冲图重印Clark等，2010年6月 ，麦克金尼等。。 2010 14，克拉克和快速，2011年16麦克金尼和克拉克，按14。 <img alt="图7" src="/files/ftp_upload/3387/3387fig7.jpg" /> 图7。电机的变化诱发电位的ES肌肉配对脉冲TMS的大小。来自竖脊肌的肌肉和测量短的时间间隔intracortical抑制（SICI）和intracortical便利（ICF）的肌电图的痕迹的例子。 戈斯等图再版。 2011 8。 <img alt="图8" src="/files/ftp_upload/3387/3387fig8.jpg" /> 图8电机EVO的变化 。KED配对脉冲TMS的潜力大小。长间隔intracortical抑制的测量（LICI）。 LICI两个量化的测试脉冲刺激间隔100毫秒交付。第二MEP结果抑制比较第一MEP。 Clark等重印图，2010年6月 ，麦克金尼等。 2010年7麦克金尼和克拉克，在出版社14。

没有利益冲突的声明。

这篇文章的总体目标是提供了我们的实验室，视觉帐户使用经颅磁刺激的科学家和临床医生。然而，除了提供这些实验的可视化，下面我们讨论执行TMS以这种方式时要考虑的基本问题，提供了一个简要介绍TMS的反应的生理，并讨论关于使用TMS的使用别人。 一般的问题要知道当执行条所述的TMS 在执行配对脉冲TMS的时需要注意的几个问题。例如，Magstim BiStim <su...

<table border="1"><tbody><tr><th>设备名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td> 2002年经颅磁刺激 经颅磁刺激双向Stim2 图八70毫米线圈 双锥形线圈</td><td> Magstim公司</td><td>不适用</td><td> TMS设备（包括线圈） </td></tr><tr><td> Biodex系统4 </td><td> Biodex </td><td>不适用</td><td>测力计</td></tr><tr><td> BIOPAC MP150数据采集系统</td><td> BIOPAC </td><td> MP150WSW </td><td> AD转换器，肌电图和力</td></tr><tr><td> AcqKnowledge 4.0数据采集软件</td><td> BIOPAC </td><td> ACK100W </td><td></td></tr><tr><td> Nikomed跟踪1心电电极</td><td> Nikomed </td><td> 2015 </td><td>肌电图电极</td></tr><tr><td>恒定电流刺激器</td><td> Digitimer </td><td> DS7A </td><td>周围神经刺激</td></tr></tbody></table>

utilizing transcranial magnetic stimulation to study the human neuromuscular system

经颅磁刺激（TMS）已经在使用20年以上1，并已越来越流行指数在过去的十年。虽然使用TMS的扩大，在此期间，许多系统和过程的研究，原来的应用程序，或许TMS的最常见的用途之一，涉及到学习的生理，可塑性和人类的神经肌肉系统的功能。单脉冲TMS适用于运动皮层兴奋的锥体神经元transsynaptically 2（图1）和可衡量的肌电反应，可用于研究和评估人类的3皮质脊髓束的完整性和兴奋结果。此外，在磁刺激技术的最新进展现在允许皮层与脊髓兴奋4,5分区。例如，双脉冲TMS的可以用来评估intracortical易化和抑制性能相结合的条件ING刺激和刺激测试，在不同的interstimulus 时间间隔3,4,6-8。在这个视频文章中，我们将演示这些技术的方法和技术方面。具体来说，我们将证明单脉冲和双脉冲TMS的技术，适用于桡侧腕屈（FCR）的肌肉以及竖脊肌（ES）肌肉。我们的实验室研究FCR的肌肉，因为它是利益上腕手的石膏固定减少肌肉性能 6,9影响的研究，我们研究的ES肌肉由于这些肌肉的临床意义，因为它涉及到腰痛8。这说明，我们应该注意到，TMS已被用来研究的手，手臂和腿的许多肌肉，并应重申我们在FCR和ES肌肉群的游行示威，只选择了TMS的例子被用来研究人类神经肌肉制度。

Watch this Scientific Journal Video about Utilizing Transcranial Magnetic Stimulation to Study the Human Neuromuscular System at JoVE.com

Utilizing Transcranial Magnetic Stimulation to Study the Human Neuromuscular System

经颅磁刺激（TMS）是一种非侵入性的工具获得洞察力和人体神经系统的生理功能。在这里，我们目前TMS的技术研究上肢和腰部肌肉的皮质兴奋。

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 ...

utilizing-transcranial-magnetic-stimulation-to-study-human

Research

JoVE Journal

Medicine

26.4K 次观看.  Ohio University. 经颅磁刺激（TMS）是一种非侵入性的工具获得洞察力和人体神经系统的生理功能。在这里，我们目前TMS的技术研究上肢和腰部肌肉的皮质兴奋。

视频: Utilizing Transcranial Magnetic Stimulation to Study the Human Neuromuscular System

A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional capacity. Transcutaneous surface stimulation of skeletal muscle involves a current flow between a cathode and an anode, thereby inducing excitement of the motor unit and the surrounding muscle fibers. 
NMES is an attractive modality to evaluate skeletal muscle adaptive responses for several reasons. First, it provides a reproducible experimental model in which physiological adaptations, such as myofiber hypertophy and muscle strengthening6, angiogenesis7-9, growth factor secretion9-11, and muscle precursor cell activation12 are well documented. Such physiological responses may be carefully titrated using different parameters of stimulation (for Cochrane review, see 13). In addition, NMES recruits motor units non-selectively, and in a spatially fixed and temporally synchronous manner14, offering the advantage of exerting a treatment effect on all fibers, regardless of fiber type. Although there are specified contraindications to NMES in clinical populations, including peripheral venous disorders or malignancy, for example, NMES is safe and feasible, even for those who are ill and/or bedridden and for populations in which rigorous exercise may be challenging. 
Here, we demonstrate the protocol for adapting commercially available electrodes and performing a NMES protocol using a murine model. This animal model has the advantage of utilizing a clinically available device and providing instant feedback regarding positioning of the electrode to elicit the desired muscle contractile effect. For the purpose of this manuscript, we will describe the protocol for muscle stimulation of the anterior compartment muscles of a mouse hindlimb.

A murine model of neuromuscular electrical stimulation (NMES), a safe and inexpensive clinical modality, to the anterior compartment muscles is described. This model has the advantage of modifying a readily available clinical device for the purpose of eliciting targeted and specific muscle contractions in mice.

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 ...

科研

医学

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human malignancies.1 Other than surgery for the minority of patients who present with localized disease, there is little or no survival benefit of systemic therapy. Therefore, there is a great need to better understand the biology of NETs, and in particular define new therapeutic targets for patients with nonresectable or metastatic neuroendocrine tumors. 3D cell culture is becoming a popular method for drug screening due to its relevance in modeling the in vivo tumor tissue organization and microenvironment.2,3 The 3D multicellular spheroids could provide valuable information in a more timely and less expensive manner than directly proceeding from 2D cell culture experiments to animal (murine) models.
To facilitate the discovery of new therapeutics for NET patients, we have developed an in vitro 3D multicellular spheroids model using the human NET cell lines. The NET cells are plated in a non-adhesive agarose-coated 24-well plate and incubated under physiological conditions (5% CO2, 37 &deg;C) with a very slow agitation for 16-24 hr after plating. The cells form multicellular spheroids starting on the 3rd or 4th day. The spheroids become more spherical by the 6th day, at which point the drug treatments are initiated. The efficacy of the drug treatments on the NET spheroids is monitored based on the morphology, shape and size of the spheroids with a phase-contrast light microscope. The size of the spheroids is estimated automatically using a custom-developed MATLAB program based on an active contour algorithm. Further, we demonstrate a simple method to process the HistoGel embedding on these 3D spheroids, allowing the use of standard histological and immunohistochemical techniques. 
This is the first report on generating 3D spheroids using NET cell lines to examine the effect of therapeutic drugs. We have also performed histology on these 3D spheroids, and displayed an example of a single drug's effect on growth and proliferation of the NET spheroids. Our results support that the NET spheroids are valuable for further studies of NET biology and drug development.

We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy.

人类神经内分泌肿瘤细胞株为研究人类神经内分泌肿瘤治疗的三维模型

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

Utilizing Repetitive Transcranial Magnetic Stimulation to Improve Language Function in Stroke Patients with Chronic Non-fluent Aphasia

Transcranial magnetic stimulation (TMS) has been shown to significantly improve language function in patients with non-fluent aphasia1. In this experiment, we demonstrate the administration of low-frequency repetitive TMS (rTMS) to an optimal stimulation site in the right hemisphere in patients with chronic non-fluent aphasia. A battery of standardized language measures is administered in order to assess baseline performance. Patients are subsequently randomized to either receive real rTMS or initial sham stimulation. Patients in the real stimulation undergo a site-finding phase, comprised of a series of six rTMS sessions administered over five days; stimulation is delivered to a different site in the right frontal lobe during each of these sessions. Each site-finding session consists of 600 pulses of 1 Hz rTMS, preceded and followed by a picture-naming task. By comparing the degree of transient change in naming ability elicited by stimulation of candidate sites, we are able to locate the area of optimal response for each individual patient. We then administer rTMS to this site during the treatment phase. During treatment, patients undergo a total of ten days of stimulation over the span of two weeks; each session is comprised of 20 min of 1 Hz rTMS delivered at 90% resting motor threshold. Stimulation is paired with an fMRI-naming task on the first and last days of treatment. After the treatment phase is complete, the language battery obtained at baseline is repeated two and six months following stimulation in order to identify rTMS-induced changes in performance. The fMRI-naming task is also repeated two and six months following treatment. Patients who are randomized to the sham arm of the study undergo sham site-finding, sham treatment, fMRI-naming studies, and repeat language testing two months after completing sham treatment. Sham patients then cross over into the real stimulation arm, completing real site-finding, real treatment, fMRI, and two- and six-month post-stimulation language testing.

We explore the use of repetitive transcranial magnetic stimulation (rTMS) to improve language abilities in patients with chronic stroke and non-fluent aphasia. After identifying a site in the right frontal gyrus for each patient that responds optimally to stimulation, we target this site during ten days of rTMS treatment.

利用重复经颅磁刺激改善脑卒中患者的语言功能与慢性非流利性失语

Transcranial magnetic stimulation (TMS) has been shown to significantly improve language function in patients with non-fluent aphasia1. In this ...

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including Parkinson's and Huntington's can disrupt it. People with Parkinson's disease, for example, tend to have increased saccadic latencies. Consequently, the quantitative measurement of saccadic eye movements has received considerable attention as a potential biomarker for neurodegenerative conditions. A lot more can be learned about the brain in both health and disease by observing what happens to eye movements when the function of specific brain areas is perturbed. Deep brain stimulation is a surgical intervention used for the management of a range of neurological conditions including Parkinson's disease, in which stimulating electrodes are placed in specific brain areas including several sites in the basal ganglia. Eye movement measurements can then be made with the stimulator systems both off and on and the results compared. With suitable experimental design, this approach can be used to study the pathophysiology of the disease being treated, the mechanism by which DBS exerts it beneficial effects, and even aspects of normal neurophysiology.

This paper describes the use of quantitative measurement of eye movements in conjunction with stimulation of focal areas of the deep brain in order to study physiology, pathophysiology, and the mechanisms of deep brain stimulation.

使用Saccadometry与深部脑刺激来研究正常和病理脑功能

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including ...

Using Retinal Imaging to Study Dementia

The retina offers a unique &#8220;window&#8221; to study pathophysiological processes of dementia in the brain, as it is an extension of the central nervous system (CNS) and shares prominent similarities with the brain in terms of embryological origin, anatomical features and physiological properties.&#160; The vascular and neuronal structure in the retina can now be visualized easily and non-invasively using retinal imaging techniques, including fundus photography and optical coherence tomography (OCT), and quantified semi-automatically using computer-assisted analysis programs. Studying the associations between vascular and neuronal changes in the retina and dementia could improve our understanding of dementia and, potentially, aid in diagnosis and risk assessment.&#160; This protocol aims to describe a method of quantifying and analyzing retinal vasculature and neuronal structure, which are potentially associated with dementia. This protocol also provides examples of retinal changes in subjects with dementia, and discusses technical issues and current limitations of retinal imaging.

The retina shares prominent similarities with the brain and thus represents a unique window to study vasculature and neuronal structure in the brain non-invasively. This protocol describes a method to study dementia using retinal imaging techniques. This method can potentially aid in diagnosis and risk assessment of dementia.

利用视网膜成像研究痴呆

The retina offers a unique &#8220;window&#8221; to study pathophysiological processes of dementia in the brain, as it is an extension of the central ...

Surgical Labyrinthectomy of the Rat to Study the Vestibular System

To study the vestibular system or the vestibular compensation process, a number of methods have been developed to cause vestibular damage, including surgical or chemical labyrinthectomy and vestibular neurectomy. Surgical labyrinthectomy is a relatively simple, reliable, and rapid method. Here, we describe the surgical technique for rat labyrinthectomy. A postauricular incision is made under general anesthesia to expose the external auditory canal and the tympanic membrane, after which the tympanic membrane and the ossicles are removed without the stapes. The stapes artery, which is located between the stapes and the oval window, is a vulnerable structure and must be preserved to obtain a clear surgical field. A hole to fenestrate the vestibule is made with a 2.1-mm drill bur superior to the stapes. Then, 100% ethanol is injected through this hole and aspirated several times. Meticulous dissection under a microscope and careful bleeding control are essential to obtain reliable results. Symptoms of vestibular loss, such as nystagmus, head tilting, and a rolling motion, are seen immediately after surgery. The rotarod or rotation chair test can be used to objectively and quantitatively evaluate the vestibular function.

This protocol describes the surgical labyrinthectomy of a rat, which is a useful method for studying the vestibular system.

大鼠前庭系统的外科迷路切除术研究

To study the vestibular system or the vestibular compensation process, a number of methods have been developed to cause vestibular damage, including ...

Updated Technique for Reliable, Easy, and Tolerated Transcranial Electrical Stimulation Including Transcranial Direct Current Stimulation

Transcranial direct current stimulation (tDCS) is a noninvasive method of neuromodulation using low-intensity direct electrical currents. This method of brain stimulation presents several potential advantages compared to other techniques, as it is noninvasive, cost-effective, broadly deployable, and well-tolerated provided proper equipment and protocols are administered. Even though tDCS is apparently simple to perform, correct administration of the tDCS session, especially the electrode positioning and preparation, is vital for ensuring reproducibility and tolerability. The electrode positioning and preparation steps are traditionally also the most time consuming and error-prone. To address these challenges, modern tDCS techniques, using fixed-position headgear and pre-assembled sponge electrodes, reduce complexity and setup time while also ensuring that the electrodes are consistently placed as intended. These modern tDCS methods present advantages for research, clinic, and remote-supervised (at home) settings. This article provides a comprehensive step-by-step guide for administering a tDCS session using fixed-position headgear and pre-assembled sponge electrodes. This guide demonstrates tDCS using commonly applied montages intended for motor cortex and dorsolateral prefrontal cortex (DLPFC) stimulation. As described, selection of the head size and montage-specific headgear automates electrode positioning. Fully assembled pre-saturated snap-electrodes are simply affixed to the set position snap-connectors on the headgear. The modern tDCS method is shown to reduce setup time and reduce errors for both novice and expert operators. The methods outlined in this article can be adapted to different applications of tDCS as well as other forms of transcranial electrical stimulation (tES) such as transcranial alternating current stimulation (tACS) and transcranial random noise stimulation (tRNS). However, since tES is application specific, as appropriate, any methods recipe is customized to accommodate subject, indication, environment, and outcome specific features.

When administering transcranial direct current stimulation (tDCS), reproducible electrode preparation and placement are vital for a tolerated and effective session. The purpose of this article is to demonstrate updated modern setup procedures for the administration of tDCS and related transcranial electrical stimulation techniques, such as transcranial alternating current stimulation (tACS).

用于可靠、简单和耐受的颅内电刺激（包括颅内直流刺激）的更新技术

Transcranial direct current stimulation (tDCS) is a noninvasive method of neuromodulation using low-intensity direct electrical currents. This method of ...

Visualization of Endogenous Mitophagy Complexes In Situ in Human Pancreatic Beta Cells Utilizing Proximity Ligation Assay

Mitophagy is an essential mitochondrial quality control pathway, which is crucial for pancreatic islet beta cell bioenergetics to fuel glucose-stimulated insulin release. Assessment of mitophagy is challenging and often requires genetic reporters or multiple complementary techniques not easily utilized in tissue samples, such as primary human pancreatic islets. Here we demonstrate a robust approach to visualize and quantify formation of key endogenous mitophagy complexes in primary human pancreatic islets. Utilizing the sensitive proximity ligation assay technique to detect interaction of the mitophagy regulators NRDP1 and USP8, we are able to specifically quantify formation of essential mitophagy complexes in situ. By coupling this approach to counterstaining for the transcription factor PDX1, we can quantify mitophagy complexes, and the factors that can impair mitophagy, specifically within beta cells. The methodology we describe overcomes the need for large quantities of cellular extracts required for other protein-protein interaction studies, such as immunoprecipitation (IP) or mass spectrometry, and is ideal for precious human islet&#160;samples generally not available in sufficient quantities for these approaches. Further, this methodology obviates the need for flow sorting techniques to purify beta cells from a heterogeneous islet population for downstream protein applications. Thus, we describe a valuable protocol for visualization of mitophagy highly compatible for use in heterogeneous and limited cell populations.

This protocol outlines a method for quantitative analysis of mitophagy protein complex formation specifically in beta cells from primary human islet samples. This technique thus allows analysis of mitophagy from limited biological material, which are crucial in precious human pancreatic beta cell samples.

利用邻近结扎测定法在人类胰腺β细胞中原位内源性米托巴结物的可视化

Mitophagy is an essential mitochondrial quality control pathway, which is crucial for pancreatic islet beta cell bioenergetics to fuel glucose-stimulated ...

Employing the Forced Oscillation Technique for the Assessment of Respiratory Mechanics in Adults

There is increasing interest in the use of the forced oscillation technique (FOT) or oscillometry to characterize respiratory mechanics in healthy and diseased individuals. FOT, a complementary method to traditional pulmonary function testing, utilizes a range of oscillatory frequencies superimposed on tidal breathing to measure the functional relationship between airway pressure and flow. This passive assessment provides an estimate of respiratory system resistance (Rrs) and reactance (Xrs) that reflect airway caliber and energy storage and dissipation, respectively. Despite the recent increase in popularity and updated Technical Standards, clinical adoption has been slow which relates, in part, to the lack of standardization regarding the acquisition and reporting of FOT data. The goal of this article is to address the lack of standardization across laboratories by providing a comprehensive written protocol for FOT and an accompanying video. To illustrate that this protocol can be utilized irrespective of a particular device, three separate FOT devices have been employed in the case examples and video demonstration. This effort is intended to standardize the use and interpretation of FOT, provide practical suggestions, as well as highlight future questions that need to be addressed.

As the use of forced oscillation technique (FOT) is increasingly utilized to characterize respiratory mechanics, there is a need to standardize methods with respect to nascent technical guidelines and various manufacturer's recommendations. A detailed protocol is provided including FOT assessment and interpretation for two cases to facilitate the standardization of methods.

采用强制振荡技术评估成人呼吸力学

There is increasing interest in the use of the forced oscillation technique (FOT) or oscillometry to characterize respiratory mechanics in healthy and ...

Study Design for Navigated Repetitive Transcranial Magnetic Stimulation for Speech Cortical Mapping

The cortical areas involved in human speech should be characterized reliably prior to surgery for brain tumors or drug-resistant epilepsy. The functional mapping of language areas for surgical decision-making is usually done invasively by electrical direct cortical stimulation (DCS), which is used to identify the organization of the crucial cortical and subcortical structures within each patient. Accurate preoperative non-invasive mapping aids surgical planning, reduces time, costs, and risks in the operating room, and provides an alternative for patients not suitable for awake craniotomy. Non-invasive imaging methods like MRI, fMRI, MEG, and PET are currently applied in presurgical design and planning. Although anatomical and functional imaging can identify the brain regions involved in speech, they cannot determine whether these regions are critical for speech. Transcranial magnetic stimulation (TMS) non-invasively excites the cortical neuronal populations by means of electric field induction in the brain. When applied in its repetitive mode (rTMS) to stimulate a speech-related cortical site, it can produce speech-related errors analogous to those induced by intraoperative DCS. rTMS combined with neuronavigation (nrTMS) enables neurosurgeons to preoperatively assess where these errors occur and to plan the DCS and the operation to preserve the language function. A detailed protocol is provided here for non-invasive speech cortical mapping (SCM) using nrTMS. The proposed protocol can be modified to best fit the patient- and site-specific demands. It can also be applied to language cortical network studies in healthy subjects or in patients with diseases that are not amenable to surgery.

Navigated repetitive transcranial magnetic stimulation is a highly efficient non-invasive tool for mapping speech-related cortical areas. It helps in designing brain surgery and speeds up the direct cortical stimulation conducted during the surgery. This report describes how to perform speech cortical mapping reliably for preoperative evaluation and research.

用于语音皮质映射的导航重复经颅磁刺激的研究设计

The cortical areas involved in human speech should be characterized reliably prior to surgery for brain tumors or drug-resistant epilepsy. The functional ...