XHTW WM凯克基金会杰出青年学者在医学研究中，也是由美国国立卫生研究院/ NHLBI补助R01 - HL089598和R01HL091947和肌肉萎缩症协会授予＃69238的支持。这项工作是支持部分由基金会Leducq CaMKII信号在心脏联盟。 NL是2009-2010年的米歇尔Mirowski国际奖学金在接受心脏起搏与电从心脏节律协会和2009-2012年美国心脏协会博士后奖学金。

第1部分。手术准备<ol><li>编程的电刺激实验是在小鼠的生存期手术无菌条件下是必要的。然而，在本质上是最常见的类型的实验终端，普通明确的手术技巧不够的。 </li><li>鼠标使用2％，在0.5 L / min的100％O 2的异氟醚麻醉。 </li><li>卷发器是用来刮胡子的皮草，从领口到中期胸部水平。 </li><li>麻醉鼠标被放置在一个仰卧的姿势，其四肢录音心电图电极在加热板（梧桐仪器，休斯敦，得克萨斯州）成立上。手术区消毒，用10％聚维酮碘。建议的系统，包括直肠温度探头连接到一个加热垫，由一个thermoanalyzer系统控制，体温保持在37.0 ° C ± 1.0 ° C。 </li></ol>第2部分。插管和EP的导管插入右心房和心室<ol><li>脚趾捏鼠标完全麻醉，确认后，1 / 2英寸的切口中线右侧骶管总站与锁骨水平。 </li><li>分隔钝性分离皮下组织，唾液腺和淋巴组织，以可视化的右颈内静脉。 </li><li>与近端的静脉用6-0缝合。轻轻地拉动这个缝合，将保持直颈内静脉插入导管。另一个缝合静脉划归在可视化段的末端。这缝线将导管绑在一次心导管放置在最佳状态，以确保止血和保持在所需的位置（ 图1）导管。 </li><li>基于计算机的数据采集，现在开始记录体表心电图同时腔内电图（即IOX - 2数据采集软件，Emka技术，弗吉尼亚州，美国）和4个信息。心腔内电图记录，导管上的电极连接在“录音”模式（即STG3008模型，多通道系统，罗伊特林根，德国）的外部刺激。 </li><li>使用微型剪刀，在纵向方向的静脉，是一个小切口和先进1.1F octapolar导管（EPR - 800，米勒仪器，休斯顿，德克萨斯）是通过静脉流入右心房。轻轻拉动近端缝合，将有助于保持直颈内静脉，将允许更容易通过导管进入右心房和心室。验证正确的导管位置是由4腔内电图的波形图的可视化右心室，右心室，房室结，右心房的基地，分别为（图2）的顶点水平。 </li><li>现在与远端缝合关闭，以确保导管的位置，并防止可能发生的的出血。 </li></ol>第3部分。程控刺激<ol><li>在选择性心房调搏的情况下，位于中庭内的电极对“重新编码”模式切换“刺激”的模式，而另一个电极对留在“录音”模式。 </li><li>要确定鼠标是否有房性心律失常的脆弱性增加，右心房的电刺激进行编程。首先，心房起搏阈值确定的应用，在不同的基本周期长度（BCL）中的2毫秒（至少50个）的电流脉冲刺激捕获的一致性测试。初始脉冲列车BCL略高于内在BCL较低，是10毫秒（例如，100毫秒，90毫秒，80毫秒，70毫秒）下降。典型的电流刺激捕获所需的幅度是100-200μA。 </li><li>窦房结恢复时间（SNRT）测量后，申请在100毫秒BCL一个15秒的心房起搏列车。 SNRT最后的刺激和起搏列车第一次自发的窦性搏动的发生之间的时间间隔定义为。 </li><li>心房有效不应期（AERP）是由心房起搏列车系列应用在一个固定的BCLS一个加上较短的S2过早刺激（即100毫秒）。 S1 - S2的时间间隔逐步减少2 - MS在每个起搏列车从70ms的20ms的。 AERP定义为最长的S1 - S2的心房耦合故障间隔生成一个定期列车脉冲S2（S1和S2过早刺激）传播击败。彼此之间的刺激协议，是一个至少30秒的恢复期。 </li><li>房室的有效不应期（AV）是由节点（AVNERP）申请在100毫秒BCL耦合S2过早刺激心房起搏列车系列。 S1 - S2的时间间隔是逐步减少2 MS每个起搏列车从70毫秒到20毫秒。 AVNERP被定义为S1 - S2的耦合最长的间隔过早刺激传递到心房是由他的潜力，而不是由QRS波群。彼此之间的刺激协议，是一个至少30秒的恢复期。 </li><li>诱发房性心律失常，包括心房纤维颤动（AF），可使用Verheule 等突发起搏协议测试2 2秒阵阵应用于确定房性心律失常的诱发。第2次爆裂，一个40毫秒的周期长度（CL），和一个2 - MS CL比以前爆裂短，每次连续2秒爆裂，直到最终CL 20毫秒。在心律失常基板的情况下，心脏会立即恢复窦性心律，起搏协议。在心房扑动的情况下，将会有一个快速一个心房电图上看到的波规律，而心室反应的频率通常是慢，心室电图上看到。房颤的情况下，体表心电图P波的情况下将显示在不规则的RR间隔。此外，心房电图，就会发现一个快速和不规则波，而心室电图将揭示不规则和较慢的心室波（ 图3） 。通常心律失常感应协议一式三份，被认为是存在的，如果它可以在至少有2 3项试验诱发心律失常 。 3,4。 </li></ol>第4部分。拔除尿管<ol><li>所有起搏协议完成后，数据采集停止。在导管的末端缝合轻轻切断，以释放导管。 </li><li>在终端EP研究的情况下，结轻轻地松开释放导管。 </li><li>在研究的结论，将鼠标人道安乐死，而下异氟醚使用颈椎脱位。 </li></ol>代表性的成果体表心电图和心内电图记录同时在整个体内的电生理学研究，并完成所有协议的详细审查。基准电参数包括PQ的时间间隔，PR间期，QRS时限，QT间期，QRS形态。可以测量这些参数，手动或自动使用数据采集软件IOX - 2或心电图自动（Emka技术，弗吉尼亚州，美国）。 SNRT，AERP和AVNERP提供有关窦房结“领跑者”的功能，心房，房室结传导性能，分别的信息。起搏诱发房颤发作的例子，可以发现在纸， 车路等。 <img src="/files/ftp_upload/1730/1730fig1.jpg" alt="图1" /> 图1。腔内导尿在小鼠体内的电生理学研究的插图。右侧颈内静脉是孤立的，空心的。静脉末端打结一次取得适当的导管位置，B 。鼠标被放置在此过程中仰卧位C.卡通描绘的导管腔内的位置。对电极定位于右心室，右心室的基础，房室结，右心房的顶点水平， 分别为， D。特写1.1F octapolar导管。图Mathur 等人的修改。中国保监会Arrhtyhm Electrophys 5的许可。 <img src="/files/ftp_upload/1730/1730fig2.jpg" alt="图2" /> 图2。在鼠标的代表体表心电图和心内电图 。 （一）体表心电图显示正常窦性节律，频率为540次每分钟。（BE）的双极心内电图的录音（二）在右心房水平，房室结（三），和基地铅II配置（四）右心室，右心室（E）的顶点分别。请注意，在B组的A波腔内对应体表心电图的P波。心室电图的V型波对应体表心电图上QRS波。 <img src="/files/ftp_upload/1730/1730fig3.jpg" alt="图3" /> 图3。在鼠标后心房突发起搏房颤代表体表心电图和心内电图。

<ol>
	<li>Nattel, S., Shiroshita-Takeshita, A., Brundel, B. J., Rivard, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+atrial+fibrillation:+lessons+from+animal+models.">Mechanisms of atrial fibrillation: lessons from animal models.</a> Prog Cardiovasc Dis. 48, 9-9 (2005).</li><li>Verheule, S., Sato, T., Everett, T. t., Engle, S. K., Otten, D., Rubart-von der Lohe, M., Nakajima, H. O., Nakajima, H., Field, L. J., Olgin, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+vulnerability+to+atrial+fibrillation+in+transgenic+mice+with+selective+atrial+fibrosis+caused+by+overexpression+of+TGF-beta1.">Increased vulnerability to atrial fibrillation in transgenic mice with selective atrial fibrosis caused by overexpression of TGF-beta1.</a> Circ Res. 94, 1458-1458 (2004).</li><li>Sood, S., Chelu, M. G., Oort, R. J. v. a. n., Skapura, D., Santonastasi, M., Dobrev, D., Wehrens, X. H., H, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+calcium+leak+due+to+FKBP12.6+deficiency+in+mice+facilitates+the+inducibility+of+atrial+fibrillation.">Intracellular calcium leak due to FKBP12.6 deficiency in mice facilitates the inducibility of atrial fibrillation.</a> Heart Rhythm. 5, 1047-1047 (2008).</li><li>Chelu, M. G., Sarma, S., Sood, S., Wang, S., van Oort, R. J., Skapura, D. G., Li, N., Santonastasi, M., Muller, F. U., Schmitz, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calmodulin+kinase+II-mediated+sarcoplasmic+reticulum+Ca2++leak+promotes+atrial+fibrillation+in+mice.">Calmodulin kinase II-mediated sarcoplasmic reticulum Ca2+ leak promotes atrial fibrillation in mice.</a> J Clin Invest. 119, 1940-1940 (2009).</li><li>Mathur, N., Subeena, S., Wang, S., van Oort, R. J., Sarma, S., Li, N., Skapura, D., Bayle, J. H., Valderrabano, M., Wehrens, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sudden+Infant+Death+Syndrome+in+Mice+With+an+Inherited+Mutation+in+RyR.">Sudden Infant Death Syndrome in Mice With an Inherited Mutation in RyR.</a> Circ Arrhythmia Electrophysiol. , (2009).</li></ol>

在心脏导管插入术，导尿过程中的广泛出血，可能会增加心率由于血容量不足。在这种情况下，腹腔注射无菌生理盐水（0.3 1.0毫升）可以正常化充盈压，减少鼠标的血流动力学压力。 吸入异氟醚的时间超过2小时，或异氟醚的浓度较高（&gt; 2％），能抑制在小鼠的心脏和呼吸功能。因此，建议所有的研究，在不到2个小时完成。此外，它是必不可少的，体​​温始终维持在正?...

programmed electrical stimulation in mice

转基因小鼠已成为一个较好的动物模型，研究底层的传导异常，心房和心室心律不齐，心脏猝死的分子机制。<sup&gt; 1</sup&gt;心内起搏研究可以进行小鼠使用1.1​​F octapolar的导管插入颈静脉，流入右心房和心室先进。在这里，我们说明了在执行编程小鼠电刺激所涉及的步骤。体表心电图和心内电图同步记录在心房，房室交界区，心肌，而心内起搏的心房使用一个外部的刺激。因此，程控电刺激的小鼠提供了独特的机会，发掘潜在的传导缺陷和心律失常的分子机制。

Watch this Scientific Journal Video about Programmed Electrical Stimulation in Mice at JoVE.com

Programmed Electrical Stimulation in Mice

程控电刺激的能力，以确定心脏的传导性能，并有可能诱发和终止使用各种起搏协议心律不齐。使用经静脉导管，心内电图的录音，可以得到以下编程的电刺激协议，以确定心律失常基板的小鼠。

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 ...

programmed-electrical-stimulation-in-mice

Research

JoVE Journal

Medicine

20.2K 次观看.  Baylor College of Medicine (BCM). 程控电刺激的能力，以确定心脏的传导性能，并有可能诱发和终止使用各种起搏协议心律不齐。使用经静脉导管，心内电图的录音，可以得到以下编程的电刺激协议，以确定心律失常基板的小鼠。

视频: Programmed Electrical Stimulation in Mice

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

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. ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

A Multimodal Imaging- and Stimulation-based Method of Evaluating Connectivity-related Brain Excitability in Patients with Epilepsy

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations over time in the blood-oxygenation level dependent signal. rs-fcMRI has been applied extensively to identify abnormalities in brain connectivity in different neurologic and psychiatric diseases. However, the relationship among rs-fcMRI connectivity abnormalities, brain electrophysiology and disease state is unknown, in part because the causal significance of alterations in functional connectivity in disease pathophysiology has not been established. Transcranial Magnetic Stimulation (TMS) is a technique that uses electromagnetic induction to noninvasively produce focal changes in cortical activity. When combined with electroencephalography (EEG), TMS can be used to assess the brain's response to external perturbations. Here we provide a protocol for combining rs-fcMRI, TMS and EEG to assess the physiologic significance of alterations in functional connectivity in patients with neuropsychiatric disease. We provide representative results from a previously published study in which rs-fcMRI was used to identify regions with abnormal connectivity in patients with epilepsy due to a malformation of cortical development, periventricular nodular heterotopia (PNH). Stimulation in patients with epilepsy resulted in abnormal TMS-evoked EEG activity relative to stimulation of the same sites in matched healthy control patients, with an abnormal increase in the late component of the TMS-evoked potential, consistent with cortical hyperexcitability. This abnormality was specific to regions with abnormal resting-state functional connectivity. Electrical source analysis in a subject with previously recorded seizures demonstrated that the origin of the abnormal TMS-evoked activity co-localized with the seizure-onset zone, suggesting the presence of an epileptogenic circuit. These results demonstrate how rs-fcMRI, TMS and EEG can be utilized together to identify and understand the physiological significance of abnormal brain connectivity in human diseases.

Resting-state functional-connectivity MRI has identified abnormalities in patients with a wide range of neuropsychiatric disorders, including epilepsy due to malformations of cortical development. Transcranial Magnetic Stimulation in combination with EEG can demonstrate that patients with epilepsy have cortical hyperexcitability in regions with abnormal connectivity.

评价中的连接有关的大脑兴奋患者癫痫多式联运Imaging-和激励为基础的方法

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations ...

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 ...

评估颅脑电刺激对纤维肌痛的疗效

A Randomized, Sham-Controlled Trial of Cranial Electrical Stimulation for Fibromyalgia Pain and Physical Function, Using Brain Imaging Biomarkers

Fibromyalgia is a chronic pain syndrome that presents with a constellation of broad symptoms, including decreased physical function, fatigue, cognitive disturbances, and other somatic complaints. Available therapies are often insufficient in treating symptoms, with inadequate pain control commonly leading to opioid usage for attempted management. Cranial electrical stimulation (CES) is a promising non-pharmacologic treatment option for pain conditions that uses pulsed electrical current stimulation to modify&#160;brain function via transcutaneous electrodes. These neural mechanisms and the applications of CES in fibromyalgia symptom relief require further exploration.
A total of 50 participants from the Atlanta Veterans Affairs Healthcare System (VAHCS) diagnosed with fibromyalgia were enrolled and then block-randomized into either a placebo plus standard therapy or active CES plus standard therapy group. Baseline assessments were obtained prior to the start of treatment. Both interventions occurred over 12 weeks, and participants were assessed at 6 weeks and 12 weeks after treatment initiation. The primary outcome investigated whether pain and functional improvements occur with the application of CES. Additionally, baseline and follow-up resting state functional connectivity magnetic resonance imaging (rs-fcMRI) were obtained at the 6-week and 12-week time points to assess for clinical applications of neural connectivity biomarkers and the underlying neural associations related to treatment effects.
This is a randomized, placebo-controlled trial to determine the efficacy of CES for improving pain and function in fibromyalgia and further develop rs-fcMRI as a clinical tool to assess the neural correlates and mechanisms of chronic pain and analgesic response.

作者聚焦：慢性疼痛管理研究的方法和进展

The current study is a randomized, placebo-controlled trial to determine the efficacy of cranial electrical stimulation (CES) for improving pain and function in fibromyalgia and further develop resting functional connectivity magnetic resonance imaging (rs-fcMRI) as a clinical tool to assess the neural correlates and mechanisms of chronic pain and analgesic response.

一项使用脑成像生物标志物进行颅脑电刺激治疗纤维肌痛疼痛和身体功能的随机、假对照试验

Fibromyalgia is a chronic pain syndrome that presents with a constellation of broad symptoms, including decreased physical function, fatigue, cognitive ...