这项工作是由Grant的奖数R01HL096593和K23HL72825从国家心脏，肺和血液研究所的支持。的内容是纯粹的托马斯杰斐逊大学的责任，并不一定代表国家心脏，肺和血液研究所和美国国立卫生研究院的官方意见。

病例报告 （如适用）:不适用，这仍然是一个实验的研究过程，尚未应用于临床。 诊断，评估和计划 （必填，如果适用）:不适用，这仍然是一个实验的研究过程，尚未应用于临床。 过程 （必填）:这部分应包括有关程序一步一步的描述，下面的指导方针。 1。毛细管显微镜（ 图1） <ol><li>我们的甲襞微循环技术被改编从Serne和他的同事们1。在此过程中的排除标准是胶原血管疾病，，因为胶原血管疾病称为毛细管变化。 </li><li>至少10小时通宵速度快，20分钟层次的休息，微血管的测量进行了一个半小时，在7上午11时，在一个安静的温控室（保持在21.5-22.5°C之间），在心脏水平的坐姿，左手的主题。 </li><li>甲襞毛细血管中的第三个手指背皮肤是采用体视显微镜（奥林巴斯中心谷，PA），与4万像素的的SPOT洞察单色数码相机（型号IN-1400:诊断仪器;斯特林高地，MI），和一台笔记本电脑（戴尔Latitude D600戴尔奥斯汀，德克萨斯州）。要限制运动，左手和前臂松散地覆盖有一个折叠的毯子，并搁在显微镜的基础上定位在另一个折起的毯子。 </li><li>甲床照明是实现一个250-W卤素光纤光纤灯（KL 2500LCD:肖特-Fostec艾姆斯佛德，纽约）;额外照明的补充150W光纤光缆卤素光源（B＆B显微镜有限公司，Warrendale，PA）的使用在深色个人。 </li><li>可视化的毛细血管，使用3.2倍物镜​​（奥林巴斯3.2/0.07）与系统总放大倍数为38.4x。 </li><li>使用相同的标准SPOT软件功能（拉伸光明和黑暗的水平），以最大限度地提高知名度的毛细血管，在所有科目使用SPOT成像相机随附的软件在毛细管的照片，光/暗对比度增强。 </li><li>量化毛细血管密度，数字显微照片，每3-5秒在每个分三个阶段，在休息的基线，在闭塞后反应性充血，并在静脉闭塞。 （一）在休息基线，显微检测毛细血管灌注在休息三分钟的时间内，被接管。 （b）在闭塞后反应性充血，显微采取量化功能灌注的毛细血管（基线加上储备毛细血管），如下所示。首先，左上臂的遮挡扎口膨胀收缩压为10分钟以上至40毫米汞柱。显微照片，然后在第E首先分钟后立即释放所有功能灌注的毛细血管动脉闭塞，可视化。反应性充血的毛细血管密度显示功能受损毛细血管招聘，和因此功能稀薄。 （c）在静脉闭塞显微照片采取量化最大的毛细血管密度，其中既包括灌注（活性红血细胞（RBC）的运动）和未注满（充满停滞，非移动的红细胞）毛细血管3如下。经过10分钟的休息后，闭塞后反应性充血过程，手臂袖口膨胀到60毫米汞柱，持续60秒，被动地迫使血液进入所有专利毛细血管目前，在这段时间内采取显微照片。由于毛细血管密度最大，包括所有的毛细血管结构，减少毛细血管密度最大表示结构疏松。 </li><li>毛细血管密度被定义为每平方密耳的毛细血管的数目limeter甲襞的皮肤，并且被计算为四个测量值的平均值，得到的四个最清晰的图像，至少扭曲运动。在我们的研究中，已被用于毛细管计数的典型值（毛细血管个/ mm 2）55-80为基线，65-90的局部缺血后，和90-105静脉闭塞。 ％毛细管征聘的值通常是在5％和25％（平均〜10-15％）之间，和％灌注毛细管之间的70％和95％（意思〜80-90％），其值较低，比正常血压的高血压。三个观察员进行独立评估的10个不同主题的照片（ 图2）手动计数过程的重现性得到了验证。观察员的身份和血压这些科目被蒙蔽。后来独立进行的计数培训，表现出高度认同。平均评分和评分间差异的奥德r的2-3毛细血管/月2，组内相关系数均大于0.90。短期的甲襞微循环变化的措施的评分和评分间差异的同一量级（约2毛细血管/毫米2），但观察到2-3年的长期的变化是一个数量级更大的（约15毛细管/毫米2），表明纵向变化，可以容易地区别于评价者变化。的毛细作用的措施的可靠性也高（组内相关系数为0.84，％的毛细招聘和0.82％的灌注毛细血管）。 </li><li>调查人员现在利用以计算机为基础的方法量化毛细血管密度使用Image-Pro Plus的成像软件（版本6.2，Media Cybernetics公司，马里兰州贝塞斯达， 图3）。基线，缺血后，和肝静脉之间的Pearson相关我们充血计数完成10个科目的软件和相应的手动计数分别为0.78，0.78和0.71​​（P &lt;0.05），表明两种方法之间的合理的协议。基于计算机的计数的可靠性是略低于手动计数，但仍处于高位（组内相关系数= 0.91为基准，静脉闭塞缺血后0.86和0.84）。我们有未公布的数据也证明了该协会的自动计数与多种心血管危险因素，包括高血压，这是我们目前正在准备出版。 </li><li> 表1总结了毛细血管密度的测定和计算。除以毛细血管密度的增加引起的闭塞后反应性充血（闭塞后反应性充血的毛细血管密度，减去基线毛细血管密度），的毛细血管密度最大的（无源静脉观察毛细管招聘百分比进行评估我们阻塞）。百分比灌注毛细血管代表所有的毛细血管目前，灌注功能活跃，比​​例的计算方法是将闭塞后反应性充血的毛细血管密度最大的毛细血管密度。两者％的毛细管招聘和百分之灌注毛细血管反映毛细血管功能。较低的值对这些措施表示功能的毛细管稀疏。 </li></ol> 2。血管内皮功能的评估<ol><li>血管内皮功能的评估之前和之后闭塞后的反应性充血，使用非侵入性的体积描记法测量前臂的血流量，根据该方法，4 Sivertsson利用内皮依赖性的刺激诱 ​​导的血管舒张反应性充血。 </li><li>平卧休息10分钟后，在坐姿的主题，水银的橡胶应变计拉长到10％，超出了它的休息长度的循环around为主体的前臂5厘米以下的肘前窝。 </li><li>应变计连接到一个体积描记器（EC-4:DE霍坎森公司;贝尔维尤，WA），而这又是连接到多普勒记录（CW-1，DE霍坎森公司; Belleveue，WA）。 </li><li>上臂闭塞袖口，和手臂被暂停使用吊​​带绷带连接到一个可调节的静脉极舒适心脏水平。收缩压和舒张压血压和心脏率DINAMAP特全公司的100自动BP袖口（GE Healthcare公司，新泽西州Piscataway，）放置在相反的手臂。 </li><li>儿科袖带膨胀到200毫米汞柱堵塞流的手绕在手腕上。上臂袖带充气至50毫米汞柱，瘪1.5秒，然后迅速重新充气之前每个前臂血流量的测量，通过扩展的应变计对周围的前臂下得到。 </li><li>前臂血流量（FBF）休息基线（FBF 基地 ）和一个测量吨闭塞后充血引起的最大血管舒张（FBF 最大 ）。对于基线血流量测量，连续四个FBF曲线是在30秒内（FBF 基地 ）。 </li><li>的遮挡扎口，然后膨胀，收缩压为10分钟以上至40毫米汞柱。释放后动脉闭塞（闭塞后反应性充血），在第一个30秒的流量（FBF 最大 ）四个连续的的FBF曲线获得。 </li><li>计算的比率FBF 最高 / FBF 碱作为血管舒张的估计值，由四个FBF 基值的平均值除以四个FBF max值的平均值。5前臂血管阻力在最大血管舒张（FVR 最大 ）被计算为平均动脉压（MAP）除以FBF 最大 。 :FBF 最大反应性充血时被直接有关FBF最大输注动脉内乙酰胆碱后，内皮依赖性的血管扩张剂。6 </sup&gt;的因此，FBF max和比FBF 最大 / FBF 基地的被接受的非侵入性的血管内皮功能的措施。6-8此外，FBF max和FVR 最大反映电阻动脉的结构变化（增加壁/管腔比）的9 </li></ol>

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没有透露这项工作的行为或发布相关的经济利益冲突。

毛细管显微镜（毛细血管显微镜）的毛细管结构是一个非标准的措施。然而，目前有没有为直接毛细管structure.Furthermore评估的标准方法，毛细血管显微镜已被广泛用于直接评价capillarydensity发表的作品，12日，13日，10日，11日，14日至18日在大，越来越多的。此外，我们已经证实了的毛细血管显微镜技术的关联甲襞微循环的研究结果与前臂血流，血管功能的一个行之有效的措施，10日，11日和阿联酋，血管损伤（未发表的作品，手稿，准备一套行之有效的措施） 。而原来的方法，利用连续的可视化，使用视频摄影，我们的简化方法利用反复分钟拍摄的数字照片，详细的程序段中。 尽管理论上真实峰值毛细血管密度共同ULD使用静止图像数毛细血管不需要一个运行的录像带是指被低估或遗漏，我们利用1分钟快速连续地拍摄的静止图像，在繁忙的反应性充血以下版本的袖口，特别是计数的前四个由此获得的图像，提供最大的缺血后微血管计数。此外，有报道称，通过使用长时间的上臂缺血在suprasystolic压力为几分钟，充血的反应，可能会持续3至4分钟，在健康受试者至少，作为只有10秒后，1分钟堵塞相对手指套19 我们做了适应的技术等Serne。有两个原因:1）主要使该技术更实际和在临床和临床研究设置用于访问，和2）为了更密切镜像的方法，我们已经使用验证技术COMPA响我们的研究结果发现在前臂plethymography的技术，这是一个建立的方法评估血管功能的那些。我们使用了一个手臂袖口数字袖口的地方，这是不容易在美国。另外，作为上面提到的，与数字缺血上臂的使用可能导致更长时间的充血响应，贷款本身更容易捕捉到的峰值充血响应分析。缺血结果在更大的充血流量（产生较高的肱动脉血管舒张。在发展中的技术，而不是前臂，上臂袖带的位置，我们的研究文献中动脉闭塞最适当的时间，我们发现期间从3分钟19日至5分钟4，6，8至13分钟，20，21与一个使用13分钟的研究报告重现性好，由此得到的值，然而，另一个研究表明增加血流量与INCreasing闭塞时间22因此，我们选择了10分钟为一个中间值。 我们引进了新的参数， 百分比毛细管招聘 ，这表示毛细管招聘的最大数量的百分比显示在静脉闭塞，在试图招募血管正常化的人数除以总数的毛细血管目前，中，能够直接比较积极灌流（功能）毛细血管个人之间的总数。我们也报告了越来越广泛的应用参数（毛细管招募和静脉闭塞）在我们的出版物的日期。 我们的技术采用了低倍率（38.4 X）相比，在以前的文献报道的其他。毛细血管密度詹姆斯/邵氏等 23（98-117每平方毫米 ）和安东尼奥斯12，谁也使用高倍率196x（57 - 93％的报告0.68毫米2）确实较高。然而，Debbabi /利维24日获得毛细血管密度（60-79）我们（55-74）几乎是相同的，使用200倍的放大倍率。我们的数量实际上是更高比那些报道由Serne 等。25，谁用100倍的放大倍率（48-57）。这可以是至少部分地由于船只每场的绝对数量减少的事实，即在较高的放大倍数，使之更难以识别数目减少了的血管的疾病。此外，我们使用的38.4x放大产生的数字图像，从中我们可以得到几个相邻的4，而不需要进一步的处理（缩小或放大）的图像清晰的领域。 在我们的研究中，我们已选择研究毛细血管末梢循环，这是使用一个简单的体视显微镜在指尖易于接触，并可以很容易地进行研究，而无需使用静脉内染料注射26 </s&gt;据指出，甲襞微循环的技术是很难进行高色素的主题24，27，没有毛细血管稀疏的黑色主题报告之前，我们的研究结果。使用双光源和照片增强软件中描述的方法，我们可以在所有受试可视化和量化的毛细血管，包括深色的黑人。 综上所述，毛细管显微镜是一种非侵入性的，相对廉价的直接可视化的微循环的方法。甲襞微循环和前臂技术，易于接受的患者，可以学得很快。的微血管和内皮细胞的功能，使用本文介绍的方法可能有未来效用在临床病人的心血管疾病的危险减贫战略措施。正如我们所发表的报告表明，微血管和内皮功能障碍的初始阶段hypertension包括高血压前期，甲襞微循环和静脉闭塞体积描记法，最终可能会有助于早期识别，风险分层和预防终末期血管病变，其潜在的致命后果。

non-invasive assessment of microvascular and endothelial function

作者们利用甲襞微循环和前臂血流技术调查的微血管功能障碍在心血管疾病的发病机制中的作用。毛细管显微镜是一种非侵入性的，相对廉价的直接可视化的微循环的方法。毛细管招聘百分比评估，将毛细血管密度的增加引起的闭塞后反应性充血（闭塞后反应性充血毛细血管密度减去基线毛细血管密度），最大毛细血管密度（被动静脉阻塞的过程中观察到的）。百分比灌注毛细血管代表所有的毛细血管目前，灌注功能活跃，比​​例的计算方法是将闭塞后反应性充血的毛细血管密度最大的毛细血管密度。两者％的毛细管招聘和百分之灌注毛细血管反映毛细血管功能。前臂血流（FBF）技术公关ovides接受血管内皮功能的非侵入性的措施:该比率FBF 最高 / FBF 碱作为血管舒张的估计值，计算除以四个FBF由四个FBF 基值的平均值的最大值的平均值。前臂血管阻力最大的血管扩张（FVR 最大值 ）计算的平均动脉压（MAP）除以FBF 最大 。甲襞微循环和前臂技术，易于接受的患者，可以学得很快。 的微血管和内皮细胞的功能，使用本文介绍的方法可能有未来效用在临床病人的心血管疾病的危险减贫战略措施。正如我们所发表的报告表明，微血管和内皮功能障碍被发现在初始阶段包括高血压前期，高血压，血管和内皮细胞功能的措施最终可能会有助于早期识别，终末期血管病变的危险分层和预防，其潜在的致命后果。

在血压正常和高血压个体之间的微血管的外观的差异是显而易见的，通过比较图4和图5。组织良好的行中的血压正常的个体中， 图4示出了典型的网络直毛细血管。相比之下， 图5显示了一个弄乱模式，皱缩，卷曲的毛细血管。 作者有持续的兴趣的微血管功能障碍在心血管疾病的发病机制中的作用。在他们的第一NIH / NHLBI资助，成功地组建了一个200人的队列，包括正常血压，高血压前期，高血压患者，并进行了一系列的调查，对微血管功能障碍（毛细血管疏松和血管内皮功能的措施）。在这个世代的个人年龄从18-55岁不等，60％为女性，46％的非裔美国人，61％PREHypertensive组（n = 122）。我们公布的报告微血管功能障碍被发现在初始阶段，高血压（高血压前期和一期高血压:以前出版的详细信息。10，11，虽然横截面，我们的研究结果表明，高血压血管的病理过程已经开始了在这个世代展示温和的水平，血压升高，表明微血管功能障碍可能早慢性持续性高血压的发展。 <table border="1"><tbody><tr><td>毛细血管密度=毛细管数为每平方毫米（mm 2）的手指甲襞皮肤<ol class="ualpha"><li> 休息基准:持续灌注毛细管1 </li><li> 闭塞后反应性充血性+间歇性持续灌注灌流储备功能毛细血管;措施的毛细作用1 </li><li> 静脉阻塞（MAXI正常毛细血管密度）:所有毛细管目前，最大的可视化包括灌注（活性红血细胞（RBC）的运动）和未注满（填充有停滞，非移动的红细胞）毛细管，毛细管构造体3的措施</li></ol></td></tr><tr><td> 毛细管招聘百分比 =（BA）÷C×100 [闭塞后反应性充血毛细血管密度 - 安静的基线毛细血管密度] ÷ 最大毛细血管密度（在被动静脉​​淤血） 测量的毛细作用 百分比灌注毛细血管 =（B÷C）×100 闭塞后反应性充血的毛细血管密度 ÷ 最大毛细血管密度（在被动静脉​​淤血） 测量的毛细作用 </td></tr></tbody></table> 表1。毛细结构的措施和功能。 <img alt="图1" src="/files/ftp_upload/50008/50008fig1.jpg" /> 图1。毛细管显微镜诊断毛细血管，特别是甲床，用显微镜检查。 <img alt="图2" src="/files/ftp_upload/50008/50008fig2.jpg" /> 图2。甲襞微循环图像。计数的手。盒装注意的是绘制使用一个Staedler的工程统治者，并表示1平方毫米。 <img alt="图3" src="/files/ftp_upload/50008/50008fig3.jpg" /> 图3。计算机辅助甲襞微循环的Image-Pro软件是用来提高对比度（例如在右侧）在原来的数字图像（例如在左侧），然后，可以使用该软件计数。 图4。甲襞微循环图像:血压正常的个体。 <img alt="图6" src="/files/ftp_upload/50008/50008fig5.jpg" /> 图5。甲襞微循环图像:高血压个人。

Watch this Scientific Journal Video about Non-invasive Assessment of Microvascular and Endothelial Function at JoVE.com

Non-invasive Assessment of Microvascular and Endothelial Function

毛细管显微镜是一种非侵入性的，相对廉价的直接可视化的微循环的方法。前臂血流技术提供接受血管内皮功能的非侵入性的措施。

非侵入性的评估微血管和内皮功能

The authors have utilized capillaroscopy and forearm blood flow techniques to investigate the role of microvascular dysfunction in pathogenesis of ...

non-invasive-assessment-of-microvascular-and-endothelial-function

Universidad San Sebastian

Research

JoVE Journal

Medicine

16.1K 次观看.  Thomas Jefferson University. 毛细管显微镜是一种非侵入性的，相对廉价的直接可视化的微循环的方法。前臂血流技术提供接受血管内皮功能的非侵入性的措施。

视频: Non-invasive Assessment of Microvascular and Endothelial Function

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

Perinatal brain injury remains a significant cause of infant mortality and morbidity, but there is not yet an effective bedside tool that can accurately screen for brain injury, monitor injury evolution, or assess response to therapy. The energy used by neurons is derived largely from tissue oxidative metabolism, and neural hyperactivity and cell death are reflected by corresponding changes in cerebral oxygen metabolism (CMRO2). Thus, measures of CMRO2 are reflective of neuronal viability and provide critical diagnostic information, making CMRO2 an ideal target for bedside measurement of brain health.
Brain-imaging techniques such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT) yield measures of cerebral glucose and oxygen metabolism, but these techniques require the administration of radionucleotides, so they are used in only the most acute cases.
Continuous-wave near-infrared spectroscopy (CWNIRS) provides non-invasive and non-ionizing radiation measures of hemoglobin oxygen saturation (SO2) as a surrogate for cerebral oxygen consumption. However, SO2 is less than ideal as a surrogate for cerebral oxygen metabolism as it is influenced by both oxygen delivery and consumption. Furthermore, measurements of SO2 are not sensitive enough to detect brain injury hours after the insult 1,2, because oxygen consumption and delivery reach equilibrium after acute transients 3. We investigated the possibility of using more sophisticated NIRS optical methods to quantify cerebral oxygen metabolism at the bedside in healthy and brain-injured newborns. More specifically, we combined the frequency-domain NIRS (FDNIRS) measure of SO2 with the diffuse correlation spectroscopy (DCS) measure of blood flow index (CBFi) to yield an index of CMRO2 (CMRO2i) 4,5.
With the combined FDNIRS/DCS system we are able to quantify cerebral metabolism and hemodynamics. This represents an improvement over CWNIRS for detecting brain health, brain development, and response to therapy in neonates. Moreover, this method adheres to all neonatal intensive care unit (NICU) policies on infection control and institutional policies on laser safety. Future work will seek to integrate the two instruments to reduce acquisition time at the bedside and to implement real-time feedback on data quality to reduce the rate of data rejection.

We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.

婴幼儿的脑代谢和血流动力学的非侵入式光学测量

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

科研

医学

Murine Spinotrapezius Model to Assess the Impact of Arteriolar Ligation on Microvascular Function and Remodeling

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. Its unique anatomy makes the spinotrapezius useful for investigation of ischemic injury and subsequent microvascular remodeling. Here, we demonstrate an arteriolar ligation model in the murine spinotrapezius muscle that was developed by our research team and previously published1-3. For certain vulnerable mouse strains, such as the Balb/c mouse, this ligation surgery reliably creates skeletal muscle ischemia and serves as a platform for investigating therapies that stimulate revascularization. Methods of assessment are also demonstrated, including the use of intravital and confocal microscopy. The spinotrapezius is well suited to such imaging studies due to its accessibility (superficial dorsal anatomy) and relative thinness (60-200 &mu;m). The spinotrapezius muscle can be mounted en face, facilitating imaging of whole-muscle microvascular networks without histological sectioning. We describe the use of intravital microscopy to acquire metrics following a functional vasodilation procedure; specifically, the increase in arterilar diameter as a result of muscle contraction. We also demonstrate the procedures for harvesting and fixing the tissues, a necessary precursor to immunostaining studies and the use of confocal microscopy.

We demonstrate a novel arterial ligation model in murine spinotrapezius muscle, including a step-by-step procedure and description of required instrumentation. We describe the surgery and relevant outcome measurements relating to vascular network remodeling and functional vasodilation using intravital and confocal microscopy.

小鼠Spinotrapezius模型来评估的影响小动脉结扎​​术，微血管功能与重塑

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

小鼠模型的非侵入性成像检测和监测卵巢癌复发

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

A Methodological Approach to Non-invasive Assessments of Vascular Function and Morphology

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis. Damage to the endothelium may predispose the vessel to atherosclerosis and increase the risk for cardiovascular disease. Assessments of peripheral endothelial function are good indicators of early abnormalities in the vascular wall and correlate well with assessments of coronary endothelial function. The present manuscript details the important methodological steps necessary for the assessment of microvascular endothelial function using laser Doppler imaging with iontophoresis, large vessel endothelial function using flow-mediated dilatation, and carotid atherosclerosis using carotid artery ultrasound. A discussion on the methodological considerations for each of the techniques is also presented, and recommendations are made for future research.

The present article describes the methodological considerations for several non-invasive assessments of vascular function and morphology that are commonly used in medical research to assess different stages of atherosclerosis.

方法论方法血管功能和形态的非侵入性评估

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis.  Damage to the endothelium may ...

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic mechanisms that contribute to the onset and progression of intestinal diseases. The identification of new molecules that promote these pathologies has led to a flurry of activity focused on the development of potential new therapies to inhibit their function. As a result, various pre-clinical mouse models with an intact immune system and stromal microenvironment are now heavily used. Here we describe three experimental protocols to test the efficacy of new therapeutics in pre-clinical models of (1) acute mucosal damage, (2) chronic colitis and/or colitis-associated colon cancer, and (3) sporadic colorectal cancer. We also outline procedures for serial endoscopic examination that can be used to document the therapeutic response of an individual tumor and to monitor the health of individual mice. These protocols provide complementary experimental platforms to test the effectiveness of therapeutic compounds shown to be well tolerated by mice.

We describe methods for longitudinal monitoring of the efficacy of therapeutics for the treatment of colonic pathologies in mice using a rigid endoscope. This protocol can be readily used for the characterization of the therapeutic response of an individual tumor in live mice and also for monitoring potential disease relapse.

新的治疗肠道病变的疗效，使用活体小鼠的串行内镜成像的非侵入性评估

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic ...

Transcutaneous Assessment of Renal Function in Conscious Rodents

Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and time-consuming. In addition, serial blood or urine samples are required, with the associated stress for the experimental animals. A recent technique significantly reduces the investment in time and resources, minimizing the invasiveness and the animal stress, but being equally valid as the traditional approaches. The method measures transcutaneously renal function. Using an optical device and the exogenous renal marker fluorescein isothiocyanate (FITC)-sinistrin, this technique is capable of measuring the elimination kinetics of the marker through the skin. With neither blood nor urine samples nor the associated laboratory assays needed, the results of the transcutaneous measurement are almost instantaneously available. The method has been already validated in different species and successfully applied in several models of renal pathology. Moreover, due to its minimally invasive characteristics, it is suitable for sequential measurements within the same animal. Here is provided a detailed protocol to carry out the transcutaneous assessment of renal function in rodents.

Determination of glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional procedures to measure this parameter are cumbersome and require a large investment of time. Here we describe a faster and minimally invasive method to determinate GFR transcutaneously.

在鼠害自觉肾功能的评估皮

Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and ...

Ultrasound Assessment of Endothelial Function: A Technical Guideline of the Flow-mediated Dilation Test

Cardiovascular disease is the primary cause of mortality and a major cause of disability worldwide. The dysfunction of the vascular endothelium is a pathological condition characterized mainly by a disruption in the balance between vasodilator and vasoconstrictor substances and is proposed to play an important role in the development of atherosclerotic cardiovascular disease. Therefore, a precise evaluation of endothelial function in humans represents an important tool that could help better understand the etiology of multiple cardio-centric pathologies.
Over the past twenty-five years, many methodological approaches have been developed to provide an assessment of endothelial function in humans. Introduced in 1989, the FMD test incorporates a forearm occlusion and subsequent reactive hyperemia that promotes nitric oxide production and vasodilation of the brachial artery. The FMD test is now the most widely utilized, non-invasive, ultrasonic assessment of endothelial function in humans and has been associated with future cardiovascular events.
Although the FMD test could have clinical utility, it is a physiological assessment that has inherited several confounding factors that need to be considered. This article describes a standardized protocol for determining FMD including the recommended methodology to help minimize the physiological and technical issues and improve the precision and reproducibility of the assessment.

The flow mediated dilation (FMD) test is the most commonly utilized, non-invasive, ultrasound assessment of endothelial function in humans. Although the FMD test has been related with the prediction of future cardiovascular disease and events, it is a physiological assessment with many inherent confounding factors that need to be considered.

血管内皮功能的超声评估:血流介导的舒张试验的技术指导原则

Cardiovascular disease is the primary cause of mortality and a major cause of disability worldwide. The dysfunction of the vascular endothelium is a ...

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. It is hypothesized that pro-atherogenic mediators released from fat tissues particularly in association with central/visceral adiposity may promote pathogenic vascular changes locally and systemically, and the notion that cardiovascular disease may be the consequence of adipose tissue dysfunction continues to evolve. Here, we describe a unique method of videomicroscopy that involves analysis of vasodilator and vasoconstrictor responses of intact small human arterioles removed from the adipose depot of living human subjects. Videomicroscopy is used to examine functional properties of isolated microvessels in response to pharmacological or physiological stimuli using a pressured system that mimics in vivo conditions. The technique is a useful approach to gain understanding of the pathophysiology and molecular mechanisms that contribute to vascular dysfunction locally within the adipose tissue milieu. Moreover, abnormalities in the adipose tissue microvasculature have also been linked with systemic diseases. We applied this technique to examine depot-specific vascular responses in obese subjects. We assessed endothelium-dependent vasodilation to both increased flow and acetylcholine in adipose arterioles (50 - 350 &#181;m internal diameter, 2 - 3 mm in length) isolated from two different adipose depots during bariatric surgery from the same individual. We demonstrated that arterioles from visceral fat exhibit impaired endothelium-dependent vasodilation compared to vessels isolated from the subcutaneous depot. The findings suggest that the visceral microenvironment is associated with vascular endothelial dysfunction which may be relevant to clinical observation linking increased visceral adiposity to systemic disease mechanisms. The videomicroscopy technique can be used to examine vascular phenotypes from different fat depots as well as compare findings across individuals with different degrees of obesity and metabolic dysfunction. The method can also be used to examine vascular responses longitudinally in response to clinical interventions.

Videomicroscopy systems are used to examine functional properties of isolated adipose tissue arterioles in response to physiological and pharmacological stimuli. This technique can be used to examine microvascular phenotypes in different adipose tissue domains in obese humans.

应用 Videomicroscopy 评价人脂肪组织微血管功能

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. ...

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively affect skeletal muscle. This can result in a loss of muscle mass (atrophy) and/or loss of muscle quality (reduced force per unit of muscle mass), both of which are prevalent in chronic disease, muscle-specific disease, immobilization, and aging (sarcopenia). Skeletal muscle function in animals can be evaluated by a range of different tests. All tests have limitations related to the physiological testing environment, and the selection of a specific test often depends on the nature of the experiments. Here, we describe an in vivo, non-invasive technique involving a helpful and easy assessment of force frequency-curve (FFC) in mice that can be performed on the same animal over time. This permits monitoring of disease progression and/or efficacy&#160;of a potential therapeutic treatment.

Measurement of rodent skeletal muscle contractile function is a useful tool that can be used to track disease progression as well as efficacy of therapeutic intervention. We describe here the non-invasive, in vivo assessment of the dorsiflexor muscles that can be repeated over time in the same mouse.

小鼠肌功能的无创评价

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively ...

A Computerized Functional Skills Assessment and Training Program Targeting Technology Based Everyday Functional Skills

Today, many functional skills are technology-based, so development of a technology-based training program has broad importance. Here we present a computerized functional skills training program that was paired in half of the participants with a commercially available cognitive training (CCT) program.
Non-impaired older individuals (NC) aged 60+ (n=45) and similarly aged individuals with mild cognitive impairment (MCI; n=50) were randomized to receive 12 weeks of twice-weekly computerized functional skills training (CFST) or 12 weeks of twice-weekly sessions split between CCT and CFST. Skills trained were use of an ATM; internet banking; ticket kiosk; telephone and internet prescription refill; medication management; and internet shopping. As with previous functional capacity assessments, we focus on completion time for each simulation.
51 participants completed the training program, either by mastering all 6 tasks (34) or completing 12 weeks of training. 44 more participants completed 4 or more training sessions so they were also analyzed for improvement up to their last training session. Completion time for all 6 tests significantly improved from the baseline assessment to the final training session in both groups of participants (all p&#60;0.001 with an average improvement in task completion time of 45%). Further, there was no differential improvement in MCI and NC in the 6 tests from baseline to end of training (all t&#60;1.66, all p&#62;0.12). Finally, combined CCT plus CFST did not differ from CSFT alone on any of the percent-change score measures (all t&#60;1.64, all p&#62;0.11).
Both NC and MCI groups evidenced substantial improvements in performance. CCT supplementation led to similar functional gains with half as many training sessions. The NC participants proceeded through the training fairly rapidly even without CCT supplementation; MCI participants required more training but learned equivalently. These findings suggest that even in cases with memory impairments, functional skills can be efficiently learned with training.

This training protocol uses computerized training to teach technology-related everyday functional skills. These skills include financial skills, travel and transit, as well as medication management.