We acknowledge funding support by K23 AT008477 (Kong), NIH K23 DA031808 (Johnson), and the Chris Redlich Endowment in Pain Research (Mackey, Dixon).

1.时间总和协议<ol><li>设备<ol><li>热脉冲传递和地点: <ol><li>使用圆形热电极，直径为2.9厘米这个热电极具有驱动的微处理器，自动化控制系统，并且，能够迅速的温度变化的珀尔帖元件。放置在受试者的鱼际热电极，并使用手套以固定的位置。 </li></ol></li><li>计算机接口: <ol><li>接口热电极上能精确控制温度交付一台笔记本电脑的软件程序。根据表1中所列的规格预编程的适当热刺激的协议的各个阶段（训练1和仅2中，作为最优化和最终的试验遵循相同的脉冲结构作为训练2）。 </li><li>通过选择合适的软件，每个阶段的热刺激计划执行热脉冲的传递，调节温度和打击"ST艺术"在用户界面上。 </li></ol></li><li>疼痛分级设备: <ol><li>使用电脑视觉模拟评分（科瓦斯）。 注:科瓦斯是表示视觉模拟评分法（VAS）单杠上移动杠杆的盒子。它是一个附属于主热电极。 </li></ol></li></ol></li><li>训练<ol><li>上定义科瓦斯视觉模拟评分（VAS）: <ol><li>目前科瓦斯的参与者和定义对VAS锚固点:0表示没有疼痛，10表示最剧烈的疼痛可想而知。确定该参与者为舒适左右移动自如的杠杆。 </li></ol></li><li>定义第二疼痛: <ol><li>指导参与者把重点放在第二疼痛从热脉冲。定义第二疼痛为"一个缓慢的，灼热，疼痛的疼痛大约需要热脉冲后的一个第二位。" </li></ol></li><li>从单一的热脉冲（训练试验1）率第二疼痛:<ol><li>在固定参与者的鱼际的热电极。指导参与者使用科瓦斯从通过热电极每个交付的单脉率第二疼痛。 </li><li>通过点击软件菜单上的"开始"按钮，开始在笔记本电脑上的预编程测试。每个加热脉冲持续时间0.5秒具有10秒（ 参照图1）的刺激间间隔。顺序地增加每表2中的加热脉冲的基线和峰温度。 </li><li>密切观察参与者的评级的第二疼痛。只要参与率的第二次痛苦比2/10以上，按软件上的停止按钮停止训练试验1。取下热电极，记录最后的热脉冲参数（ 即基线和峰值温度）。 </li></ol></li><li>从TS试验（训练试验2）疼痛率持续: <ol><li>固定在对面的手热电极。通知PARTICIPA新台币它们将接受一系列的快速10脉冲。指示参与者率仅第二疼痛，而不是着眼于快热痛的感觉，马上随每个脉冲（第疼痛）。解释给参与者，该第二疼痛可能增加，减少，或保持每个脉冲之间是相同的。 </li><li>开始在笔记本电脑上预先编程的测试。使用来自上述步骤1.2.3记录最后的基准和峰值温度。保持在0.5秒的脉冲宽度，并在2秒（ 图2）的刺激间间隔。开关的手，直到参与者舒适的评价第二疼痛持续响应快速的热脉冲必要时重复这种训练试验。 </li></ol></li></ol></li><li>优化<ol><li>定义P1，P max和TS E: <ol><li>在峰值温度下的从第一脉冲递送后约2秒定义P1作为疼痛等级（参照图3）。 11 </SUP&gt; 21 注:TS E是疼痛P1和最大疼痛（P 最大 ）之间的估计差值。 </li></ol></li><li>调整脉冲温度。旨在通过根据图4顺序地调节峰和基线温度达到30和70/100之间的TS 即 注: 见图。 4为优化算法。 </li><li>安全热电极:使用外科手套，保证热电极，以参与者的鱼际。交换机的每个审判之间的手。 </li><li>提供热脉冲:指导参与者只能从热脉冲评价他们的第二疼痛。确认参加者准备开始，然后启动预编程测试的笔记本电脑。 </li><li>迭代:重复步骤1.3.2 -1.3.4直到参与者的疼痛评级为P1是&lt;50/100 VAS和TS E是VAS 30/100和70/100之间。进行不超过5更优化试验和确保有每个审判之间的3分钟休息。从最后的审判作为最后的参数采用参数。 </li></ol></li><li>最终审判<ol><li>休息的优化试验和最终试验间5分钟。重复1.3.3和1.3.4。使用上述步骤1.3.5最终的参数。有与会者休息3分钟，然后再重复。 </li></ol></li></ol> 2.空调疼痛调制协议<ol><li>概述和设备: <ol><li>测试刺激:30秒校准到6选自10疼痛（耐热-6）接触的热（由与上述相同的热电极递送）。见2.2的详细信息。 </li><li>条件刺激: <ol><li>构造使用具有穿孔的分隔壁上的一个透明的塑料盒，并用冰和水的一侧，只是水​​的其他填充它冷水浴中。轻轻搅动然后插入温度计只水侧，以确保温度为10℃是稳定的。 </li><li>如果无法获得稳定的温度，附加水电泵到一边用冰不断循环水，以保持稳定的温度。开启泵并确保水侧温度是在10℃下是稳定的。 </li></ol></li><li>时序和计算CPM: <ol><li>在调节冷水浴的最后30秒施加试验刺激参与者一次与前一次。 CPM计算在之前和期间冷浴的最后30秒采用的热刺激6的疼痛程度的差异。 </li></ol></li></ol></li><li>用于校准热6: <ol><li>通过加热快速上升估算热6: <ol><li>固定热电极到参与者的非优势手的鱼际。在32℃下开始，提高温度以每秒0.3℃的速率。 注:升速度较慢用于优先激活C纤维20，其负责CPM。 12 </li><li>停止磨片从未参加者达到热耐受性或C最高51°。指导参与者的热量斜坡期间连续打分疼痛。重复3次，并计算平均温度在那里的痛苦评级是6/10。 </li></ol></li><li>精阈值:应用热火-6和指导参与率的痛苦。如果评级是5和7/10之间，继续2.3.2。如果是&gt; 7或&lt;5，应用一系列30秒的热热刺激的参与者（每次刺激之间交替手），从44°C至49°C，并在0.5-1℃的增量变化，直到该评级是5-7之间，允许30秒每间刺激休息。 </li><li>最终化热6和预处理的评级，测试刺激: <ol><li>结束精阈值，一旦参与者的疼痛评级是5和7/10之间。注:使用此最后，区域内疼痛为预处理测试刺激的CPM的计算的评级。也，记录最终热6导致这种等级的温度。 </li></ol></li></ol></li><li> CPM协议<ol><li>前置条件测试刺激按照步骤2.2.3。 </li><li>条件刺激: <ol><li>在10℃下施加在冷水浴对侧脚2分钟。指导参与者在0，30和90 s到率由冷水浴的痛苦。 </li></ol></li><li>重复条件刺激试验过程中的刺激: <ol><li>在冷水浴的最后30秒应用从细阈值（2.2.3），以再次对同一只手的最终热6刺激。指导参加者在30秒的热刺激，这也是120秒冷水浴月底结束时率从热6的痛苦。 注:CPM计算为在疼痛分级之前报告的测试刺激的差异和冷水浴中。 </li></ol></li></ol></li></ol>

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The authors have no conflicts of interests or any financial interests to disclose.

该议定书中的关键步骤该TS协议包括按时间顺序的关键步骤如下:多步训练（使用视觉模拟评分评价疼痛，从一个单一的加热脉冲的第二疼痛评级，并从快速加热脉冲串等级第二疼痛）;脉冲温度优化;在2-3试验，优化温度取得TS。与大多数心理的措施，参加培训万分危急确保痛苦的评级是在各试验一致，并尽可能准确。优化步骤是同样重要的，其中两个基线和峰值脉冲的温度被调节，使得第一加热脉冲的评级是小于5/10，和所述近似TS为7 3之间。 CPM的关键步骤包括视觉模拟评分疼痛分级培训，获得热火-6慢热坡道，确认热-6和精细阈值，如果necess元，应用冷水浴对侧肢体远端，并在冷水浴的最后30秒重新应用证实热6。到TS协议类似，两者的训练和热刺激的个性化（热-6）是在CPM协议的关键。另外，从经验以及从文献，在冷浴的最后30秒的重复热-6刺激是关键的，并产生比较冷浴后施加热刺激的CPM更大幅值。 26然而，鉴于有些个人不能在10摄氏度容忍冷加压的全部2分钟，这可能是合理考虑条件刺激完成后立即施加测试刺激在所有个人数据收集标准化。 修改和故障排除与TS协议的最常见的问题是，得到的TS无力，这可能是由于3主要原因。第一，也是最常见的是，疼痛评级从第一热脉冲可以是如此强大，它淹没在与随后的脉冲（TS）的疼痛的任何增加的感知。尽量减少此问题的最佳方法是优化TS的大小之前遵循协议，并依次减少基线和峰刺激温度，直到第一脉冲的疼痛评级是小于5（满分10分）。第二个原因，相对于第一个，是当与会者在即使在最高温度设定在10个脉冲的结束任何感知没有痛苦。在这种情况下，可以考虑由1或2℃提高基线脉冲温度。有时，一个人可能只是有一个很难识别和评价第二疼痛，可能是由于外周和中枢的因素。未经第二疼痛可靠感知，这是非常难以捕捉的TS。在这种情况下，我们发现温度的最好的一套，一个individual可容忍和记录TS为零。 一个成功的CPM协议中最常见的障碍是热6的不稳定性和无法容忍2分钟冷浴（10℃）。使用精细阈值在当前的协议，通过调整，以解决第一个问题的热刺激温度逐步直至疼痛评分为5和7对于第二个问题之间，注意，文献表明的抑制作用从条件刺激是饱和。 27因此，即使一个人不能他或她的脚保持冷浴2分钟，应该与此强烈痛苦的冷的刺激会出现足够的CPM效果。修改协议来记录在冷水浴浸没脚的持续时间和该参与者后立即将热刺激他或她的脚从冷水浴撤出。 CPM随后的疼痛分级热刺激的疼痛评级HEA之前减去计算ŧ刺激冷水浴后立即应用（未过程中，当一般的协议表示）。 技术的局限性这种方法也不是没有局限性。第一，尽管我们最大的努力，我们不能（分别为1遗漏参与者TS和1 CPM，）征求TS和CPM的每一个人。此，部分地，可能是由于大之间的个体差异在这些参数。 5，15，16，28，29然而，成功率为94％，这比从文献中引用的50-60％的成功率更好。 22，28二，研究者应该用这种方法，因为我们使用不同的热脉冲温度每INDI产生TS产生的TS不同个人之间演绎时要谨慎维杜阿尔。因此，横截面样品中比较的TS时，应考虑这两个在TS的幅度和在用于产生它的温度的差异。个性化的TS的方法是最适合于纵向研究，其中，重点是在同一个体加班的变化。因为同样的条件刺激用于所有个体和只在个体热6的疼痛知觉的变化被记录，而不是热-6-疼痛原始分数相同关注并不适用于个性化的CPM。尽管此方法允许TS和CPM的广泛采集，它相比于使用通用参数的方法需要更多的时间。最后，该技术需要一个有经验的操作者和高级热试验机，这两者都是不立即适应繁忙临床设置实用。我们鼓励今后的努力简化的方法。 关于到现有的技术意义。/另类方法我们的个体化​​TS和CPM参数的方法旨在消除地板和天花板效应影响，由于周边散热灵敏度变化。该方法介绍了我们组的两个更广泛的捕捉和时间效率的目标公布以前的方法改善。 11，30个体化的TS和CPM的优点是捕获上升的状态和下降疼痛处理在一个宽范围的个人，由此允许使用这些参数作为合理的测量结果为纵向研究的能力。 掌握技术后，未来的应用方向或今后的研究应着重补充修改，以节省时间，在人口众多的TS和CPM数据采集表征个体这些参数谁是无痛苦VS那些历代志的范围集成电路的疼痛，以及在TS和CPM响应于除了卷起装置和DNIC特定的生理过程的多样性的相关性。

在国际疼痛研究学会（IASP）疼痛定义为"一种不愉快的感觉和情绪体验。"慢性疼痛是指疼痛持续超过6个月。慢性疼痛是在美国一个显著的问题，影响在$ 635十亿每年成本向上100多万美国成年人。 1由于疼痛的主观性质，很难让研究者和临床医生客观地衡量一个人的痛苦经历（S），因此使得它具有挑战性的评估和治疗疼痛。出于这个原因，我们开发的标准化测试，以尽可能客观地量化疼痛是很重要的。一个这样的测试是QST，其中标准化感官刺激施用到由被检者定级。 2有静态和动态QST。前者通常评估到感觉阈值或单个刺激的等级，而后者评估了响应于一些刺激。 3日前，动态QST已经获得了，因为它提供了机会来探测传入伤害性信号的中央处理越来越多的关注。 4，5，6 动态QST的两个关键组成部分是时间总和（TS）和空调疼痛调制（CPM）。时间总和指疼痛从重复的，伤害性刺激增加的感知。 TS是卷起，在脊髓神经元继发增加显示烧制由于重复的C-光纤输入的现象行为相关。 7,8一般来说，TS可以通过使用各种伤害性刺激，如热，电，和触觉方法（ 即压力或针眼）诱导的，条件是重复刺激的频率大于0.3Hz更大，C的固有频率-纤维秒。 9,10许多研究人员使用重复的热脉冲生成生产和规范有毒热刺激，因为容易的TS。 11 CPM是指现象"痛抑制痛"，其中第二次伤害性刺激的存在降低了从最初的有害刺激的疼痛感觉。 12初始有害刺激，这是之前和之后（或期间），第二刺激的应用测量，被称为测试刺激。测试激励可以是热，电或触觉。这里热刺激经常被用来作为测试刺激，因为其在调整和标准化的缓解。 13第二刺激，称为条件刺激，通常由施加到远侧末端冷或热水浴中。 13 CPM是行为的归属关系Ø˚F弥漫有毒抑制控制（DNIC），一种生理现象，其中输入来自从异位场由C-纤维介导的所有传入刺激的脑干弥漫抑制外周C纤维的结果。 12,14 虽然TS和CPM都有反映状态和中枢性疼痛处理变化的潜力，局限性同时存在。 4，5。例如，因为在个人热灵敏度大可变性，通用刺激的应用可以导致测试的个体的高达50％的缺乏的TS。 11，15同样，热测试刺激往往造成了很可能导致CPM测试不可能的，由于地板或天花板效应广泛不同的疼痛评级。 16因此，捕捉TS和CPM广义上的协议，广告甫一热刺激是需要个人。为TS，我们调整加热脉冲的温度，以便他们可以生成在疼痛评分与每个相继脉冲充分提高;而对于CPM，我们在调整热测试刺激中度疼痛（6出10），为每个单独的使有足够的疼痛分级仍可能调理刺激后应用存在。 对于所有的精神物理学检查，在适当的评级疼痛刺激的参与者的培训是这些行为测试的准确性和再现性是至关重要的。 17当多个刺激被以较快的速度呈现，并且在CPM的情况下，当两个不同的刺激被同时施加到参加者。这是TS特别相关。此外，从热脉冲的TS，它培养参与者进行评分C-纤维介导的第二疼痛（慢，燃烧，通常是普热后亮起大约1秒钟就显得尤为重要LSE），而不是最初的阵痛（由A-δ纤维介导，并与热脉冲立即来吧）。 18,19这是较少在CPM一个问题，因为刺激有（&gt; 30秒）更长和C-纤维介导的感觉会支配在这些情况下有毒感知。 19，20在下面的协议，我们将在参与者的细节适当的培训。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Medoc Pathway CHEPS system</td>
			<td>Medoc Advanced Medical Systems&#160;&#160;</td>
			<td></td>
			<td>This system includes the machine to generate contact heat (Pathway), the thermode capable of rapid temperature change (CHEPS), and the Medoc software.&#160;</td>
		</tr>
		<tr>
			<td>CoVAS accessory hardware with the CHEPS systems</td>
			<td>Medoc Advanced Medical Systems&#160;&#160;</td>
			<td></td>
			<td>This device is a Medoc accessory that allows real-time pain rating by the participant.&#160;</td>
		</tr>
		<tr>
			<td>Laptop Computer</td>
			<td>Lenovo</td>
			<td></td>
			<td>This is the computer that runs the Medoc software and communicates with the Pathway machine.&#160;</td>
		</tr>
		<tr>
			<td>Glove</td>
			<td>Kimberly-Clark</td>
			<td></td>
			<td>Gloves are used to secure the thermode on the participant&#39;s thenar eminence.&#160;</td>
		</tr>
		<tr>
			<td>Clear plastic box with a perforated dividing wall - filled with Ice and water</td>
			<td></td>
			<td></td>
			<td>This box provides the cold water bath for the CPM task.&#160;</td>
		</tr>
		<tr>
			<td>Aquarium pump&#160;</td>
			<td>Aquarium Systems Micro-Jet pump MC 450</td>
			<td></td>
			<td>This pump circulates water, to maintain stable, even temperature in the cold water bath.&#160;</td>
		</tr>
		<tr>
			<td>Infrared Thermometer</td>
			<td>Exergen Temporal Scanner, model TAT 2000&#160;</td>
			<td></td>
			<td>To monitor constantly the temperature of the water bath.&#160;</td>
		</tr>
		<tr>
			<td>Stop Watch</td>
			<td>Any handheld stop watch or stop watch built into a smartphone</td>
			<td></td>
			<td>To prompt the participant to rate pain at specific time pointds during the CPM task.&#160;</td>
		</tr>
	</tbody>
</table>

dynamic quantitative sensory testing to characterize central pain processing

Central facilitation and modulation of incoming nociceptive signals play an important role in the perception of pain. Disruption in central pain processing is present in many chronic pain conditions and can influence responses to specific therapies. Thus, the ability to precisely describe the state of central pain processing has profound clinical significance in both prognosis and prediction. Because it is not practical to record neuronal firings directly in the human spinal cord, surrogate behavior tests become an important tool to assess the state of central pain processing. Dynamic QST is one such test, and can probe both the ascending facilitation and descending modulation of incoming nociceptive signals via TS and CPM, respectively. Due to the large between-individual variability in the sensitivity to noxious signals, standardized TS and CPM tests may not yield any meaningful data in up to 50% of the population due to floor or ceiling effects. We present methodologies to individualize TS and CPM so we can capture these measures in a broader range of individuals than previously possible. We have used these methods successfully in several studies at the lab, and data from one ongoing study will be presented to demonstrate feasibility and potential applications of the methods.

在正在进行的临床试验中，我们执行患者深表型慢性轴向腰痛，我们包括动态QST作为评估的一个组成部分。下面的表3总结了其中使用上述确切TS和CPM协议的第15例患者的基线数据。需要注意的是该数据包括患者＃19，因为不是所有的19例患者招募出现了对自己的评价，由于日程安排冲突和其他情况。 图5可视地显示在TS和CPM数据侧由端来揭示这些患者中枢性疼痛处理的变化的图案。 如上图所示，采用个性化的优化方案，我们与0-10 VAS规模平均2.7获得TS在鱼际。我们能够获得TS在所有19名学员除参加1，谁是不能够可靠地识别秒OND疼痛无需从最初的阵痛由感觉所淹没。参与者13需要早退因此没有接受植物检疫任务。否则，所有的18名学员展示CPM，其中平均约为3.1。我们成功地实现在大多数的学科一定程度的TS和CPM;并且TS和CPM的幅度与从文献一致。 13，15，22 此外，如在图4中显示出，TS和CPM的同时测量可导致对患者的在中枢性疼痛处理信息的洞察力。把它放在简体而言，高TS可能表示异常增强上升便利而低CPM建议受损降痛觉传递的抑制作用。例如，参加10，显示出7.7（满分10分）的高TS，同时参加18魔trated一个基本上不存在的CPM（-0.5 / 10）。几个先驱研究表明，中枢性疼痛处理，例如不同的配置文件可以预测术后慢性疼痛的发展的不同的速率，并以对特定中枢性疼痛途径起作用的药物不同的反应。 4，23，24，25在这里此例中，这是合理的推测，参与者10可能给加巴喷丁，钙通道阻断剂响应而参与者18，谁具有受损的CPM可能向度洛西汀，血清素-去甲肾上腺素再摄取抑制剂响应。显然还需要更多的数据和研究来验证假设。介入审判正在进行中，并会评估这些背痛患者的反应真药和假电。这里介绍的个体QST方法将允许我们准确地和纵向地跟踪在TS和CPM的变化许多个人尽可能。 <img alt="图1" src="/files/ftp_upload/54452/54452fig1.jpg" /> 图1.单热脉冲用于训练试验1.每个热普乐持续0.5秒，是除了10秒从下一个热脉冲。每个脉冲的基线和峰温度根据表2逐渐增加。只要参与者感觉到任何的脉冲，达到训练试验1的终点第二疼痛而导致的第二疼痛感觉被记录，并在训练试验2.使用脉冲的温度设置<a href="http://ecsource.jove.com/files/ftp_upload/54452/54452fig1large.jpg" target="_blank">，请点击这里查看一个更大的版本这个数字。</a> <img alt="图2" src="/files/ftp_upload/54452/54452fig2.jpg" /> 图2.热脉冲串用于培训TRIAL 2，优化并最终TS试验。十热脉冲，每0.5秒长，且2秒分开，在一个标准的TS试验被递送。 <img alt="图3" src="/files/ftp_upload/54452/54452fig3.jpg" /> 图3.定义P1，P max和TSË。 P1:从第一脉冲的第二疼痛评级; {P MAX:最大额定值的整个10脉冲串的第二疼痛; TS E:时间总和的估计的大小。 <a href="http://ecsource.jove.com/files/ftp_upload/54452/54452fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/54452/54452fig4.jpg" /> 图4.算法单独优化时间总和。这里的目标是找到基线和峰值脉冲的温度导致在预计吨emporal总和（TSE）3进出10 VAS 7之间。如果TSE低于目标，用C顺序1°增加峰值和基线的温度。如果TSE是上述目标，用C顺序1°减少基线和峰值温度。上述算法之前，应先热脉冲的肯定疼痛评级是由小（0.5-1°C）的增量减少基线和峰值脉冲温度≤5。 <a href="http://ecsource.jove.com/files/ftp_upload/54452/54452fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/54452/54452fig5.jpg" /> 图5.从正在进行的临床试验TS和CPM型材患者。在第一19例来自正在进行的临床试验慢性腰痛的TS和CPM的基线测量。相对的TS和CPM幅度的各种图案揭示于百分之电位差拉尔疼痛处理。 <a href="http://ecsource.jove.com/files/ftp_upload/54452/54452fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 审讯 </td><td> 脉冲＃ </td><td> 刺激持续时间 </td><td> 峰峰值ISI </td><td> 斜率 </td><td> 基准温度 </td><td> 最高温度 </td></tr><tr><td>训练试验1 </td><td> 1个脉冲</td><td> 0.5秒</td><td> 10秒</td><td> 40℃/秒</td><td>变量（ 见表2） </td><td>变量（ 见表2） </td></tr><tr><td>训练试验2 </td><td> 10个脉冲</td><td> 0.5秒</td><td> 2秒</td><td> 40℃/秒</td><td>从训练试验1 </td><td>从训练试验1 </td></tr><tr><td>优化试验</td><td> 10个脉冲</td><td> 0.5秒</td><td> 2秒</td><td> 40℃/秒</td><td>先从训练试验1临时工</td><td>先从训练试验1临时工</td></tr><tr><td>最终审判</td><td> 10个脉冲</td><td> 0.5秒</td><td> 2秒</td><td> 40℃/秒</td><td>个性化</td><td>个性化</td></tr></tbody></table> 表1:用于TS协议热刺激的规格（包括训练试验）。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 脉冲数 </td><td> 基线温度（℃） </td><td> 峰温度（℃） </td></tr><tr><td> 1 </td><td> 36 </td><td> 45 </td></tr><tr><td> 2 </td><td> 36 </td><td> 46 </td></tr><tr><td> 3 </td><td> 37 </td><td> 47 </td></tr><tr><td> 4 </td><td> 38 </td><td> 48 </td></tr><tr><td>五</td><td> 39 </td><td> 49 </td></tr><tr><td> 6 </td><td> 40 </td><td> 50 </td></tr><tr><td> 7 </td><td> 41 </td><td> 51 </td></tr><tr><td> 8 </td><td> 42 </td><td> 51 </td></tr><tr><td> 9 </td><td> 43 </td><td> 51 </td></tr><tr><td> 10 </td><td> 44 </td><td> 51 </td></tr></tbody></table> 表2: 温度设置用于捕捉第二疼痛在训练试验1.参见图1与这些温度参数传递单热脉冲的形状。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> SUBJ ID </td><td>碱基T </td><td>山顶ŧ </td><td> TS </td><td>热6 </td><td> CPM </td></tr><tr><td></td><td>摄氏</td><td>摄氏</td><td> VAS </td><td>摄氏</td><td> VAS </td></tr><tr><td> 1 </td><td></td><td></td><td> 0 </td><td> 44 </td><td> 4.4 </td></tr><tr><td> 2 </td><td> 41 </td><td> 51 </td><td> 0.43 </td><td> 47.6 </td><td> 4.717 </td></tr><tr><td> 3 </td><td> 40 </td><td> 50 </td><td> 0.5 </td><td> 44.7 </td><td> 4.42 </td></tr><tr><td> 4 </td><td> 38 </td><td> 49 </td><td> 0.45 </td><td> 44 </td><td> 4.355 </td></tr><tr><td>五</td><td> 42 </td><td> 51 </td><td> 3.287 </td><td> 44 </td><td> 4.0713 </td></tr><tr><td> 6 </td><td> 44 </td><td> 51 </td><td> 0.415 </td><td> 45.5 </td><td> 4.5085 </td></tr><tr><td> 7 </td><td> 44 </td><td> 51 </td><td> 4.2495 </td><td> 45.5 </td><td> 4.12505 </td></tr><tr><td> 8 </td><td> 41 </td><td> 51 </td><td> 2.575 </td><td> 45 </td><td> 4.2425 </td></tr><tr><td> 9 </td><td> 44 </td><td> 51 </td><td> 3.435 </td><td> 48 </td><td> 4.4565 </td></tr><tr><td> 10 </td><td> 43 </td><td> 51 </td><td> 7.713 </td><td> 44 </td><td> 3.6287 </td></tr><tr><td> 11 </td><td> 39 </td><td> 51 </td><td> 4.59 </td><td> 42.5 </td><td> 3.791 </td></tr><tr><td> 12 </td><td> 39 </td><td> 49 </td><td> 5.395 </td><td> 43.3 </td><td> 3.7905 </td></tr><tr><td> 13 </td><td> 44 </td><td> 51 </td><td> 2.165 </td><td></td><td> 0 </td></tr><tr><td> 14 </td><td> 39 </td><td> 49 </td><td> 2.55 </td><td> 44.1 </td><td> 4.155 </td></tr><tr><td> 15 </td><td> 40 </td><td> 51 </td><td> 4.0635 </td><td> 44 </td><td> 3.99365 </td></tr><tr><td> 16 </td><td> 42 </td><td> 51 </td><td> 3.45665 </td><td> 45.5 </td><td> 4.204335 </td></tr><tr><td> 17 </td><td> 40 </td><td> 51 </td><td> 0.953 </td><td> 46.1 </td><td>4.5147 </td></tr><tr><td> 18 </td><td> 37.5 </td><td> 49 </td><td> 0.21 </td><td> 45.5 </td><td> 4.529 </td></tr><tr><td> 19 </td><td> 40 </td><td> 51 </td><td> 2.2135 </td><td> 46.6 </td><td> 4.43865 </td></tr></tbody></table> 表3.从正在进行的临床试验TS和CPM的结果。参见图4为TS和CPM从并排绘制边露出痛苦调制谱相同的参与者。

Watch this Scientific Journal Video about Dynamic Quantitative Sensory Testing to Characterize Central Pain Processing at JoVE.com

Dynamic Quantitative Sensory Testing to Characterize Central Pain Processing

By assessing pain in response to repetitive or different types of standardized stimuli, dynamic quantitative sensory testing (QST) can reveal changes in the central processing of pain. We present methods to optimize and individualize two dynamic QST measures: temporal summation (TS) and conditioned pain modulation (CPM).

动态定量感觉测试表征中央疼痛处理

Central facilitation and modulation of incoming nociceptive signals play an important role in the perception of pain. Disruption in central pain ...

dynamic-quantitative-sensory-testing-to-characterize-central-pain

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

16.8K 次观看.  Stanford University School of Medicine. By assessing pain in response to repetitive or different types of standardized stimuli, dynamic quantitative sensory testing (QST) can reveal changes in the central processing of pain. We present methods to optimize and individualize two dynamic QST measures: temporal summation (TS) and conditioned pain modulation (CPM).

视频: Dynamic Quantitative Sensory Testing to Characterize Central Pain Processing

Physiological, Morphological and Neurochemical Characterization of Neurons Modulated by Movement

The role of individual neurons and their function in neuronal circuits is fundamental to understanding the neuronal mechanisms of sensory and motor functions. Most investigations of sensorimotor mechanisms rely on either examination of neurons while an animal is static1,2 or record extracellular neuronal activity during a movement.3,4 While these studies have provided the fundamental background for sensorimotor function, they either do not evaluate functional information which occurs during a movement or are limited in their ability to fully characterize the anatomy, physiology and neurochemical phenotype of the neuron. A technique is shown here which allows extensive characterization of individual neurons during an in vivo movement. This technique can be used not only to study primary afferent neurons but also to characterize motoneurons and sensorimotor interneurons. Initially the response of a single neuron is recorded using electrophysiological methods during various movements of the mandible followed by determination of the receptive field for the neuron. A neuronal tracer is then intracellularly injected into the neuron and the brain is processed so that the neuron can be visualized with light, electron or confocal microscopy (Fig. 1). The detailed morphology of the characterized neuron is then reconstructed so that neuronal morphology can be correlated with the physiological response of the neuron (Figs. 2,3). In this communication important key details and tips for successful implementation of this technique are provided. Valuable additional information can be determined for the neuron under study by combining this method with other techniques. Retrograde neuronal labeling can be used to determine neurons with which the labeled neuron synapses; thus allowing detailed determination of neuronal circuitry. Immunocytochemistry can be combined with this method to examine neurotransmitters within the labeled neuron and to determine the chemical phenotypes of neurons with which the labeled neuron synapses. The labeled neuron can also be processed for electron microscopy to determine the ultrastructural features and microcircuitry of the labeled neuron. Overall this technique is a powerful method to thoroughly characterize neurons during in vivo movement thus allowing substantial insight into the role of the neuron in sensorimotor function.

A technique is described to quantify the in vivo physiological response of mammalian neurons during movement and correlate the physiology of the neuron with neuronal morphology, neurochemical phenotype and synaptic microcircuitry.

由运动神经元调制生理，形态学和神经化学的表征

The role of individual neurons and their function in neuronal circuits is fundamental to understanding the neuronal mechanisms of sensory and motor ...

科研

神经科学

An Injury Paradigm to Investigate Central Nervous System Repair in Drosophila

An experimental method has been developed to investigate the cellular responses to central nervous system (CNS) injury using the fruit-fly Drosophila. Understanding repair and regeneration in animals is a key question in biology. The damaged human CNS does not regenerate, and understanding how to promote the regeneration is one of main goals of medical neuroscience. The powerful genetic toolkit of Drosophila can be used to tackle the problem of CNS regeneration.
A lesion to the CNS ventral nerve cord (VNC, equivalent to the vertebrate spinal cord) is applied manually with a tungsten needle. The VNC can subsequently be filmed in time-lapse using laser scanning confocal microscopy for up to 24 hr to follow the development of the lesion over time. Alternatively, it can be cultured, then fixed and stained using immunofluorescence to visualize neuron and glial cells with confocal microscopy. Using appropriate markers, changes in cell morphology and cell state as a result of injury can be visualized. With ImageJ and purposely developed plug-ins, quantitative and statistical analyses can be carried out to measure changes in wound size over time and the effects of injury in cell proliferation and cell death. These methods allow the analysis of large sample sizes. They can be combined with the powerful genetics of Drosophila to investigate the molecular mechanisms underlying CNS regeneration and repair.

An Injury Paradigm to Investigate Central Nervous System Repair in Drosophila

An injury paradigm using the Drosophila larval ventral nerve cord to investigate central nervous system regeneration and repair is described. Stabbing followed by laser scanning confocal microscopy in time-lapse and fixed specimens, combined with quantitative analysis with purposefully developed software and genetics, are used to investigate the molecular mechanisms of CNS regeneration and repair.

调查中枢神经系统修复中受伤范式<em&gt;果蝇</em

An experimental method has been developed to investigate the cellular responses to central nervous system (CNS) injury using the fruit-fly Drosophila. ...

Direct Intraventricular Delivery of Drugs to the Rodent Central Nervous System

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses specifically on antisense oligonucleotides (ASOs), though the techniques shown in the video here can also be used to deliver a plethora of other drugs to the CNS. Antisense oligonucleotides (ASOs) have the capability to knockdown sequence-specific targets 1 as well as shift isoform ratios of specific genes 2. To achieve widespread gene knockdown or splicing in the CNS of mice, the ASOs can be delivered into the brain using two separate routes of administration, both of which we demonstrate in the video.
The first uses Alzet osmotic pumps, connected to a catheter that is surgically implanted into the lateral ventricle. This allows the ASOs to be continuously infused into the CNS for a designated period of time. The second involves a single bolus injection of a high concentration of ASO into the right lateral ventricle. Both methods use the mouse cerebral ventricular system to deliver the ASO to the entire brain and spinal cord, though depending on the needs of the study, one method may be preferred over the other.

We describe a method to target drugs to the central nervous system by either implanting a catheter or performing a bolus injection into the right lateral ventricle in mice. We focus specifically on the delivery of antisense oligonucleotides. This technique is readily adaptable to other drugs and to rats.

啮齿动物的中枢神经系统药物直接脑室交货

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses ...

Novel Metrics to Characterize Embryonic Elongation of the Nematode Caenorhabditis elegans

Dissecting the signaling pathways that control the alteration of morphogenic processes during embryonic development requires robust and sensitive metrics. Embryonic elongation of the nematode Caenorhabditis elegans is a late developmental stage consisting of the elongation of the embryo along its longitudinal axis. This developmental stage is controlled by intercellular communication between hypodermal cells and underlying body-wall muscles. These signaling mechanisms control the morphology of hypodermal cells by remodeling the cytoskeleton and the cell-cell junctions. Measurement of embryonic lethality and developmental arrest at larval stages as well as alteration of cytoskeleton and cell-cell adhesion structures in hypodermal and muscle cells are classical phenotypes that have been used for more than 25 years to dissect these signaling pathways. Recent studies required the development of novel metrics specifically targeting either early or late elongation and characterizing morphogenic defects along the antero-posterior axis of the embryo. Here, we provide detailed protocols enabling the accurate measurement of the length and the width of the elongating embryos as well as the length of synchronized larvae. These methods constitute useful tools to identify genes controlling elongation, to assess whether these genes control both early and late phases of this stage and are required evenly along the antero-posterior axis of the embryo.

Novel Metrics to Characterize Embryonic Elongation of the Nematode Caenorhabditis elegans

Here we detail protocols specifically designed to monitor morphogenic defects that occur during early and late phases of embryonic elongation of the nematode Caenorhabditis elegans. Ultimately, these protocols are designed to identify genes that regulate these phases and to characterize their differential requirements along the antero-posterior axis of the embryo.

新颖的指标来表征线虫胚胎伸长率<em&gt;秀丽隐杆线虫</em

Dissecting the signaling pathways that control the alteration of morphogenic processes during embryonic development requires robust and sensitive metrics. ...

In Vivo Gene Transfer to Schwann Cells in the Rodent Sciatic Nerve by Electroporation

The formation of the myelin sheath by Schwann cells (SCs) is essential for rapid conduction of nerve impulses along axons in the peripheral nervous system. SC-selective genetic manipulation in living animals is a powerful technique for studying the molecular and cellular mechanisms of SC myelination and demyelination in vivo. While knockout/knockin and transgenic mice are powerful tools for studying SC biology, these methods are costly and time consuming. Viral vector-mediated transgene introduction into the sciatic nerve is a simpler and less laborious method. However, viral methods have limitations, such as toxicity, transgene size constraints, and infectivity restricted to certain developmental stages. Here, we describe a new method that allows selective transfection of myelinating SCs in the rodent sciatic nerve using electroporation. By applying electric pulses to the sciatic nerve at the site of plasmid DNA injection, genes of interest can be easily silenced or overexpressed in SCs in both neonatal and more mature animals. Furthermore, this in vivo electroporation method allows for highly efficient simultaneous expression of multiple transgenes. Our novel technique should enable researchers to efficiently manipulate SC gene expression, and facilitate studies on SC development and function.

In Vivo Gene Transfer to Schwann Cells in the Rodent Sciatic Nerve by Electroporation

Here, we present an in vivo technique for gene transfer to Schwann cells (SCs) in the rodent sciatic nerve. This simple technique is useful for investigating signaling mechanisms involved in the development and maintenance of myelinating SCs.

体内基因转移到雪旺细胞在啮齿动物坐骨神经电穿孔

The formation of the myelin sheath by Schwann cells (SCs) is essential for rapid conduction of nerve impulses along axons in the peripheral nervous ...

Physiological Preparation of Hair Cells from the Sacculus of the American Bullfrog (Rana catesbeiana)

The study of hearing and balance rests upon insights drawn from biophysical studies of model systems. One such model, the sacculus of the American bullfrog, has become a mainstay of auditory and vestibular research. Studies of this organ have revealed how sensory cells hair can actively detect signals from the environment. Because of these studies, we now better understand the mechanical gating and localization of a hair cell&#39;s transduction channels, calcium&#39;s role in mechanical adaptation, and the identity of hair cell currents. This highly accessible organ continues to provide insight into the workings of hair cells. Here we describe the preparation of the bullfrog&#39;s sacculus for biophysical studies on its hair cells. We include the complete dissection procedure and provide specific protocols for the preparation of the sacculus in specific contexts. We additionally include representative results using this preparation, including the calculation of a hair bundle&#39;s instantaneous force-displacement relation and measurement of a bundle&#39;s spontaneous oscillation.

Physiological Preparation of Hair Cells from the Sacculus of the American Bullfrog Rana catesbeiana

The American bullfrog&#39;s (Rana catesbeiana) sacculus permits direct examination of hair-cell physiology. Here the dissection and preparation of the bullfrog&#39;s sacculus for biophysical studies is described. We show representative experiments from these hair cells, including the calculation of a bundle&#39;s force-displacement relation and measurement of its unforced motion.

从美国牛蛙的小囊（毛细胞的生理准备<em&gt;牛蛙</em&gt;）

The study of hearing and balance rests upon insights drawn from biophysical studies of model systems. One such model, the sacculus of the American ...

Stimulus-specific Cortical Visual Evoked Potential Morphological Patterns

This paper presents a methodology for the recording and analysis of cortical visual evoked potentials (CVEPs) in response to various visual stimuli using 128-channel high-density electroencephalography (EEG). The specific aim of the described stimuli and analyses is to examine whether it is feasible to replicate previously reported CVEP morphological patterns elicited by an apparent motion stimulus, designed to simultaneously stimulate both ventral and dorsal central visual networks, using object and motion stimuli designed to separately stimulate ventral and dorsal visual cortical networks.&#160; Four visual paradigms are presented: 1. Randomized visual objects with consistent temporal presentation. 2. Randomized visual objects with inconsistent temporal presentation (or jitter).&#160; 3. Visual motion via a radial field of coherent central dot motion without jitter.&#160; 4. Visual motion via a radial field of coherent central dot motion with jitter.&#160; These four paradigms are presented in a pseudo-randomized order for each participant.&#160; Jitter is introduced in order to view how possible anticipatory-related effects may affect the morphology of the object-onset and motion-onset CVEP response.&#160; EEG data analyses are described in detail, including steps of data exportation from and importation to signal processing platforms, bad channel identification&#160;and removal, artifact rejection, averaging, and categorization of average CVEP morphological pattern type based upon latency ranges of component peaks. Representative data show that the methodological approach is indeed sensitive in eliciting differential object-onset and motion-onset CVEP morphological patterns and may, therefore, be useful in addressing the larger research aim. Given the high temporal resolution of EEG and the possible application of high-density EEG in source localization analyses, this protocol is ideal for the investigation of distinct CVEP morphological patterns and the underlying neural mechanisms which generate these differential responses.&#160; &#160; &#160;&#160;

In this paper, we present a protocol to investigate differential cortical visual evoked potential morphological patterns through stimulation of ventral and dorsal networks using high-density EEG. Visual object and motion stimulus paradigms, with and without temporal jitter, are described. Visual evoked potential morphological analyses are also outlined.&#160;&#160;

刺激特定的皮质视觉引发潜在形态模式

This paper presents a methodology for the recording and analysis of cortical visual evoked potentials (CVEPs) in response to various visual stimuli using ...

Advanced Diffusion Imaging in The Hippocampus of Rats with Mild Traumatic Brain Injury

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous variability and heterogeneity (age, gender, type of trauma, other possible pathologies, etc.), animal models play a key role in unraveling factors that are limitations in clinical research. They provide a standardized and controlled setting to investigate the biological mechanisms of injury and repair following TBI. However, not all animal models mimic the diffuse and subtle nature of mTBI effectively. For example, the commonly used controlled cortical impact (CCI) and lateral fluid percussion injury (LFPI) models make use of a craniotomy to expose the brain and induce widespread focal trauma, which are not commonly seen in mTBI. Therefore, these experimental models are not valid to mimic mTBI. Thus, an appropriate model should be used to investigate mTBI. The Marmarou weight drop model for rats induces similar microstructural alterations and cognitive impairments as seen in patients who sustain mild trauma; therefore, this model was selected for this protocol. Conventional computed tomography and magnetic resonance imaging (MRI) scans commonly show no damage following a mild injury, because mTBI induces often only subtle and diffuse injuries. With diffusion weighted MRI, it is possible to investigate microstructural properties of brain tissue, which can provide more insight into the microscopic alterations following mild trauma. Therefore, the goal of this study is to obtain quantitative information of a selected region-of-interest (i.e., hippocampus) to follow up disease progression after obtaining a mild and diffuse brain injury.

The overall goal of this procedure is to obtain quantitative microstructural information of the hippocampus in a rat with mild traumatic brain injury. This is done using an advanced diffusion-weighted magnetic resonance imaging protocol and region-of-interest based analysis of parametric diffusion maps.

轻度创伤性脑损伤大鼠海马学的高级扩散成像

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous ...

Assessment of Sensory Thresholds in Dogs Using Mechanical and Hot Thermal Quantitative Sensory Testing

Quantitative sensory testing (QST) is used to evaluate the function of the somatosensory system in dogs by assessing the response to applied mechanical and thermal stimuli. QST is used to determine normal dogs&#39; sensory thresholds and evaluate alterations in peripheral and central sensory pathways caused by various disease states, including osteoarthritis, spinal cord injury, and cranial cruciate ligament rupture. Mechanical sensory thresholds are measured by electronic von Frey anesthesiometers and pressure algometers. They are determined as the force at which the dog exhibits a response indicating conscious stimulus perception. Hot thermal sensory thresholds are the latency to respond to a fixed or ramped temperature stimulus applied by a contact thermode.
Following a consistent protocol for performing QST and paying attention to details of the testing environment, procedure, and individual study subjects are critical for obtaining accurate QST results for dogs. Protocols for the standardized collection of QST data in dogs have not been described in detail. QST should be performed in a quiet, distraction-free environment that is comfortable for the dog, the QST operator, and the handler. Ensuring that the dog is calm, relaxed, and properly positioned for each measurement helps produce reliable, consistent responses to the stimuli and makes the testing process more manageable. The QST operator and handler should be familiar and comfortable with handling dogs and interpreting dogs&#39; behavioral responses to potentially painful stimuli to determine the endpoint of testing, reduce stress, and maintain safety during the testing process.

This work describes a standard protocol for mechanical and hot thermal quantitative sensory testing to evaluate the somatosensory system in dogs. Sensory thresholds are measured using an electronic von Frey anesthesiometer, pressure algometer, and hot contact thermode.

使用机械和热热定量感官测试评估狗的感官阈值

Quantitative sensory testing (QST) is used to evaluate the function of the somatosensory system in dogs by assessing the response to applied mechanical ...

Dural Stimulation and Periorbital von Frey Testing in Mice As a Preclinical Model of Headache

The cranial meninges, comprised of the dura mater, arachnoid, and pia mater, are thought to primarily serve structural functions for the nervous system. For example, they protect the brain from the skull and anchor/organize the vascular and neuronal supply of the cortex. However, the meninges are also implicated in nervous system disorders such as migraine, where the pain experienced during a migraine is attributed to local sterile inflammation and subsequent activation of local nociceptive afferents. Of the layers in the meninges, the dura mater is of particular interest in the pathophysiology of migraines. It is highly vascularized, harbors local nociceptive neurons, and is home to a diverse array of resident cells such as immune cells. Subtle changes in the local meningeal microenvironment may lead to activation and sensitization of dural perivascular nociceptors, thus leading to migraine pain. Studies have sought to address how dural afferents become activated/sensitized by using either in vivo electrophysiology, imaging techniques, or behavioral models, but these commonly require very invasive surgeries. This protocol presents a method for comparatively non-invasive application of compounds on the dura mater in mice and a suitable method for measuring headache-like tactile sensitivity using periorbital von Frey testing following dural stimulation. This method maintains the integrity of the dura and skull and reduces confounding effects from invasive techniques by injecting substances through a 0.65 mm modified cannula at the junction of unfused sagittal and lambdoid sutures. This preclinical model will allow researchers to investigate a wide range of dural stimuli and their role in the pathological progression of migraine, such as nociceptor activation, immune cell activation, vascular changes, and pain behaviors, all while maintaining injury-free conditions to the skull and meninges.

The most notable symptom of migraine is severe head pain, and it is hypothesized that this is mediated by sensory neurons innervating the meninges. Here, we present a method to locally apply substances to the dura in a minimally invasive manner while using facial hypersensitivity as an output.

小鼠硬脑膜刺激和眶周冯弗雷测试作为头痛的临床前模型

The cranial meninges, comprised of the dura mater, arachnoid, and pia mater, are thought to primarily serve structural functions for the nervous system.  ...

动态定量感觉测试表征中央疼痛处理

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