由美国国立卫生研究院奖励RO1 HL073087，通用汽车公司077185，和通用汽车069589到中正支持。这项工作也从再生医学的DHLRI计划的拨款支持。作者感谢的技术支持和临床投入在美国俄亥俄州立大学的下列人员：（综合伤口中心）博士Sabyasachi.Biswas，博士阿利森Spiwak（循环技术部），约瑟夫Agoston（流通部），托马Shives（流通部），约瑟夫尤因（机械工程），斯科特Killinger（机械工程），和小尹戈（电气工程）。

1。多光谱成像的测绘组织氧合双模成像系统全面的伤口中心（“禁止化学武器公约”），从此称为“化学武器公约”制度，是发达国家在俄亥俄州立大学的多光谱成像，皮肤组织氧合和血管功能的热成像成像。较大差距二阶导数光谱技术用于重建组织氧合图1多光谱图像。在这项研究中，一个健康的主题坐在左前臂搁在台面上。一个前臂的“化学武器公约”制度领域内的部分涂上墨汁（1％溶解于乙醇），模仿不同的肤色。多光谱图像被收购，充氧地图较大差距的二阶导数谱的基础上重建。重建后的充氧地图相比，与商业Hypermed OxyVu光谱组织氧合测量系统获取。 关于同一主题的闭塞后反应性充血（PORH）2协议后，血管闭塞组织对氧的反应进行了研究。 PORH测试之前，主体的收缩压和舒张压均录得左上臂置于袖带。 PORH协议包括一个闭塞的基准期的前两分钟，一个suprasystolic闭塞（收缩压50毫米汞柱）的两分钟内，两分钟的充血和无功期间。多光谱图像被收购PORH测试期间，在每个波长为0.75秒的采样率在4个波长（即530nm，550nm处，570nm处，和590nm）。深部组织的血氧饱和度和相同的手臂上的皮肤组织的氧张力，同时OxiplexTS组织分光光度计（ISS公司，Urbana Champaign分校，IL）和中医经皮血氧监视器（辐射计，丹麦）分别录得。 2。动态热成像成像测绘血管功能动态热成像成像表现出健康的使用相同的“化学武器公约”系统的主体。舒适的主题谎称桌子上，在仰卧位，左手臂搁在台面上和背部朝向的“化学武器公约”系统的红外摄像机单元的左手。激光多普勒探头放在指尖上的同一只手的手指皮肤灌注的连续监测。一个袖带置于左上臂产生不同层次的闭塞。实验前，主题是要求要休息至少10分钟，记录的血压袖带与收缩压和舒张压。动态热成像图像被抓获以下袖压力水平：没有闭塞，0.5 ×舒张压，0.5 ×（舒张压+收缩压），以及1.5 ×收缩压。在每个袖口压力水平，介绍了30秒的左手放在一个水袋室温（25℃）热刺激。热刺激去除后，立即被收购，左手热成像图像2帧/秒的速度和手指皮肤灌注激光多普勒探头在10赫兹的采样率记录。测试之间的时间间隔为10分钟。左手的位置显着提前，使随后的测量，在相同的位置。通过计算温度响应和血管功能指数的组织血管闭塞水平在不同的功能进行了评价。热刺激后的时间和相应的温度变化之间的平方根的比值定义为血管功能指数。 3。组织氧合和血管功能的地图之间的合作注册动态热成像成像和皮肤组织氧合多光谱成像收购地图收购的皮肤组织血管功能的地图，共注册了一个先进的成像算法。四个受托标记，同时热和光的对比，被放置在生物组织，以方便合作登记的形象。目前的合作注册的光学图像与热图像（和血管功能的地图）（氧合图）协议包括以下主要步骤：1）改造中的所有图像灰度和正常化的像素点的亮度值介于0和1，2，光学照片），确定前景（皮肤）地区有一个经验性的全局阈值（对整个图像的平均强度值的0.8倍）的高像素比强度值;在前台区域的小孔填充形态的关闭操作; 3）确定作为受托标记较深的地区的前景，其强度值既克以下地区<html> lobal阈值和自适应的局部阈值平均在20 - 20像素的邻域像素值定义。自适应局部阈值，让我们以适应光照变化; 4）完善受托标记使用形态学运算去除噪声和尖峰的地区;四个地区的重心控制点的光学照片; 5）重复类似的步骤识别标记的地区，在热图像; 6）符合两套控制点的基础上接近（因为这两个相机密切定位）; 7）计算两个图像之间的仿射变换热图像作为参考和改造光学照片和氧气的地图（这是由同一摄像机的光学照片获得）相应; 8）最终生成可视化的叠加图像。 4。代表性的成果： 充氧协议（即，＃1）具有代表性的结果是皮肤组织氧合地图较大差距二阶导数光谱的基础上重建。较大差距二阶导数光谱的方法，有效地减少了测量组织的背景吸收引起的，使皮肤充氧测量模拟皮肤颜色的变化影响较小的工件。我们也表现出了PORH协议皮肤组织氧合图，深部组织的氧合，和皮肤组织的氧张力，同时记录。 血管功能的协议（即，＃2）代表性的成果，包括皮肤组织的温度分布，在应对外部热刺激皮肤组织温度的变化，以及皮肤组织血管指数动态热成像成像所得的地图。皮肤组织血管指数和皮肤组织灌注的激光多普勒测量的热成像成像之间的相关性观察。 共同注册协议（即，＃3）具有代表性的结果，包括组织氧合图，血管指数图，摄影，充氧之间的图像融合，血管指数多对比受托标记注册的图像。 <img src="/files/ftp_upload/2095/2095fig1.jpg" alt="图1" /> 图1。 “化学武器公约”双模组织氧合和血管功能的非接触式成像系统设置系统安装在移动车。它包括一个红外热成像摄像机，CCD相机，液晶可调谐滤波器，和一个宽带光源。一台计算机，用于同步​​数据的收集，分析，和显示任务。 <img src="/files/ftp_upload/2095/2095fig2.jpg" alt="图2" /> 图2。 “化学武器公约”的系统软件界面软件界面编程在LabVIEW环境。在界面左侧的顶部控制面板的硬件配置和系统校准。在界面右边是组织温度由红外摄像机收购地图的实时显示。在界面的底部，是皮肤组织灌注的激光多普勒测量。 <img src="/files/ftp_upload/2095/2095fig3.jpg" alt="图3" /> 图3。软件界面（续）“禁止化学武器公约”制度，这是一个弹出窗口显示组织氧合参数。在左边的是从多光谱图像重建的组织氧合图。界面右边显示从自上而下的组织氧合以下参数：（1）皮肤组织氧合从一个人们感兴趣的某些区域（ROI），在组织氧合图的平均;（2）深部组织的氧合OxplexTS组织血氧饱和度监测（3）皮肤组织氧分压监测中医经皮血氧监视器。 <img src="/files/ftp_upload/2095/2095fig4.jpg" alt="图4" /> 图4。皮肤组织氧合OxyVu成像系统的成像 。左边的图是一个灰度图像的皮肤组织。涂在皮肤上的油墨层来模拟皮肤的颜色。 OxyVu系统取得皮肤组织充氧的地图显示在右边。选择墨水绘制的区域内部和外部两个方方正正的感兴趣区域（投资回报）​​。墨水绘制的区域内的投资回报率，平均皮肤组织氧合62.8 ± 11.0％。墨水绘制的区域外的投资回报率，平均皮肤组织氧合44.0 ± 11.0％。 18.8％的差异，观察皮肤组织内部和外部的油墨画皮面积氧合测量。 <html>古热5“/&gt; 图5。在相同的皮肤位置就在图4中的相同主题的“化学武器公约”制度，“化学武器公约”制度的皮肤组织氧合的成像，多光谱图像被抓获。左图是皮肤组织的单一波长的图像，右图是重建皮肤组织氧合图。一层的墨水被涂在皮肤上用墨水在图4中相同的浓度。 。选择墨水绘制的区域内部和外部两个方方正正的感兴趣区域（投资回报）​​。墨水绘制的区域内的投资回报率，平均皮肤组织氧合60.9 ± 6.9％。墨水绘制的区域外的投资回报率，平均皮肤组织氧合65.8 ± 5.5％。 4.9％的差异，观察皮肤组织内部和外部的油墨画皮面积氧合测量。 <img src="/files/ftp_upload/2095/2095fig6.jpg" alt="图6" /> 图6。对皮肤组织氧合测量的可靠性的效果模拟皮肤颜色的变化进行统计分析，以确定OxyVu和“化学武器公约”的成像系统模拟皮肤颜色的变化氧合测量的意义。对于每一个成像系统，分别计算平均的皮肤组织oxygenations随机选择10地区的利益（投资回报）​​，在每个区域内部和外部墨水画皮地区。我们的零假设是，皮色不变，不会影响皮肤组织氧合测量。这个零假设进行了测试，分别使用充氧地图（即OxyVu测量）在图4和图5（即“化学武器公约”的测量）。学生的t -检验表明，OxyVu测量的P值远小于0.001，这意味着零假设被拒绝。因此，皮色不变，不影响皮肤的组织氧合测量OxyVu成像系统。相比之下，“化学武器公约”测量的P值是0.728，暗示的可能性，皮色不变，不影响皮肤组织氧合在“禁止化学武器公约”的成像系统测量。 <img src="/files/ftp_upload/2095/2095fig7.jpg" alt="图7" /> 图7。皮肤组织氧合从闭塞后反应性充血（PORH）在“化学武器公约”制度获得的图像 。PORH测试进行了前臂上一个健康的主题下面的＃1 。 （一）。单波长的灰度图像放在图像共同注册了四个受托标记的手臂。 （二）基线皮肤组织氧合图收购之前，血管闭塞。 （三）血管闭塞后的皮肤组织氧合地图（收缩压+ 50mmHg）为2分钟。 （d）在皮肤组织后，反应性充血充氧地图。在皮肤组织氧合中的明显变化之前，期间和之后，血管闭塞。 <img src="/files/ftp_upload/2095/2095fig8.jpg" alt="图8" /> 图8。组织氧参数连续监测，在一个PORH测试（一）测试协议 。显示闭塞前的基线由suprasystolic闭塞期间，反应性充血期结束期间。 （b）中绘制皮肤组织氧合期限PORH程序的历史。它获得的平均利益的选定区域（ROI），在“化学武器公约”的皮肤充氧地图。我们也OxiplexTS组织血氧饱和度监测的深部组织的氧合，（C）绘制。皮组织氧分压也中医设备和监控（d）中绘制。皮肤组织氧合的“化学武器公约”的测量与在PORH程序的其他氧气参数完全吻合。 <img src="/files/ftp_upload/2095/2095fig9.jpg" alt="图9" /> 图9。在不同的血管闭塞压力的皮肤组织的反应。热成像图像被收购后的热刺激（即，在室温25℃的水包）立即从主体的左手。横轴对应于不同的时间点被拆除后，热刺激。垂直轴对应于以下4个级别的血管闭塞压力：0（无遮挡），0.5DBP，0.5（DBP +收缩压），1.5SBP，其中舒张压舒张压和收缩压是收缩压。对于这个特定的主题，舒张压为69mmHg和收缩压是123mmHg。测试结果表明，组织的温度响应外部热刺激血管闭塞水平相关。闭塞压力增加，降低了热响应速度。 fig10.jpg“ALT =”图10“/&gt; 图10。重建在不同的血管闭塞压力的血管指数图 。血管功能指数（V）的热成像图像的每个像素是派生的，反对倒退的热刺激后时间的平方根（√吨的皮肤组织的温度变化（ΔT） ）：Δ√T + K +ε，其中ε是随机误差和K是一个常数。从左边的血管功能指数映射到正确的对应闭塞以下条件：（一）无闭塞，（二）0.5DBP，（C）0.5（DBP +收缩压），（D）1.5SBP。对于这个特定的主题，舒张压（DBP）69mmHg的收缩压（SBP）123mmHg。 <img src="/files/ftp_upload/2095/2095fig11.jpg" alt="图11" /> 图11。之间的血管功能指数和皮肤组织灌注的相关性。＃2闭塞压在每个指尖的血管功能指数计算，平均五个地区的利益，在组织中的血管指数图（投资回报）。激光多普勒设备测量指尖的皮肤组织灌注。用激光多普勒测量指尖的血管指数相关，说明组织血管功能的定量评估采用动态热成像方法的潜力。 <img src="/files/ftp_upload/2095/2095fig12.jpg" alt="图12" /> 图12。图像合作登记的过程中，在第一行中，我们显示：（一）归一的照片，（b）在分段前景的地区，和（c）在照片图像的分割标记。在第二行中，我们显示：（D）归灌注地图（E）分段前景区域，和（f）在灌注地图分段标记。两套标记位置之间的仿射变换计算。共同注册图像（G和H），得到了转化的照片图像和氧地图使用此回收的仿射变换。 <img src="/files/ftp_upload/2095/2095fig13.jpg" alt="图13" /> 图13。合作注册的结果。共转化的照片和氧气一起灌注地图地图登记，并在热图显示。热图（一）提交与转化的照片图像，红色通道的100％，100％转化氧气的形象，在绿色通道，在蓝色通道灌注地图和50％。为了更好地形象化的船只，我们目前登记结果（b）中的合作的另一个版本。热图是由在红色通道中的照片图像转化为100％，50％的转化绿色通道，氧气和50％的改变，其中血管指数图倒在蓝色通道灌注地图图像和唯一的信息前景区域内被保留。

<ol>
	<li>Myers, D. E., Anderson, L. D., Seifert, R. P., Ortner, J. P., Cooper, C. E., Beilman, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+method+for+measuring+local+hemoglobin+oxygen+saturation+in+tissue+using+wide+gap+second+derivative+near-infrared+spectroscopy.">Noninvasive method for measuring local hemoglobin oxygen saturation in tissue using wide gap second derivative near-infrared spectroscopy.</a> J Biomed Opt. 10 (3), 034017-034017 (2005).</li><li>Jarm, T., Kragelj, R., Liebert, A., Lukasiewitz, P., Erjavec, T., Preseren-Strukelj, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postocclusive+reactive+hyperemia+in+healthy+volunteers+and+patients+with+peripheral+vascular+disease+measured+by+three+noninvasive+methods.">Postocclusive reactive hyperemia in healthy volunteers and patients with peripheral vascular disease measured by three noninvasive methods.</a> Adv Exp Med Biol. 530, 661-669 (2003).</li><li>Treacher, D. F., Leach, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxygen+transport-1.+Basic+principles.">Oxygen transport-1. Basic principles.</a> Bmj. 317 (7168), 1302-1306 (1998).</li><li>Leach, R. M., Treacher, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxygen+transport-2.+Tissue+hypoxia.">Oxygen transport-2. Tissue hypoxia.</a> BMJ. 317 (7169), 1370-1373 (1998).</li><li>Sen, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+healing+essentials:+let+there+be+oxygen.">Wound healing essentials: let there be oxygen.</a> Wound Repair Regen. 17 (1), 1-18 (2009).</li><li>RL, G. r. e. e. n. m. a. n., Panasyuk, S., Wang, X., TE, L. y. o. n. s., Dinh, T., Longoria, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+changes+in+the+skin+microcirculation+and+muscle+metabolism+of+the+diabetic+foot.">Early changes in the skin microcirculation and muscle metabolism of the diabetic foot.</a> Lancet. 366 (9498), 1711-1717 (2005).</li><li>Valvano, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The use of thermal diffusivity to quantify tissue perfusion [dissertation]. , (1981).</li><li>Hassan, M., Togawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Observation+of+skin+thermal+inertia+distribution+during+reactive+hyperaemia+using+a+single-hood+measurement+system.">Observation of skin thermal inertia distribution during reactive hyperaemia using a single-hood measurement system.</a> Physiol Meas. 22 (1), 187-200 (2001).</li><li>Ley, O., Deshpande, C., Prapamcham, B., Naghavi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lumped+parameter+thermal+model+for+the+study+of+vascular+reactivity+in+the+fingertip.">Lumped parameter thermal model for the study of vascular reactivity in the fingertip.</a> J Biomech Eng. 130 (3), 031012-031012 (2008).</li><li>Wilson, S. B., Spence, V. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+thermographic+imaging+method+for+quantifying+dermal+perfusion:+potential+and+limitations.">Dynamic thermographic imaging method for quantifying dermal perfusion: potential and limitations.</a> Med Biol Eng Comput. 27 (5), 496-501 (1989).</li></ol>

氧气存在多种形式，如血红蛋白或肌红蛋白的约束的氧，溶解氧，活性氧的生物组织。氧的运输起着至关重要的作用，在保持组织的活力和正常的代谢过程3。急性轻度至中度缺氧，会主动适应代谢，血管调节，4血管生成的响应。极度缺氧，缺氧会导致足够的血管生成和细胞死亡。有限的氧气供应和增加氧气需求之间的不平衡是主要的致病因素之一，许多疾病 ，如慢性伤口5。在缺血性创面的情况下，氧气供应是有限的灌注不足，不能满足愈合过程中的代谢需要，旨在抵抗感染损害呼吸爆发功能。同时，皮肤组织氧合和血管功能的评估在慢性创面管理的临床意义。 高光谱成像技术估计照明组织皮肤组织氧合检测组织在6个不同的波长的反射率。高光谱成像的主要优势之一就是非侵入性和非接触式检测组织的功能特性。然而，许多高光谱成像系统中的氧气测量的可靠性是影响皮肤颜色和其他背景吸收变化。宽缝隙二阶导数光谱测量组织氧合以前散射和皮肤色素沉着1的影响最小。我们采用的二阶导数光谱的原则，多光谱成像和皮肤组织氧合独立与模拟皮肤颜色的变化表现出一致的测量。 组织灌注以前通过测量组织热扩散7的研究。一个单罩的方法是引入热刺激和图像组织的动态响应，以估计的皮肤组织的热惯性分布 8 。对人体前臂的皮肤受到袖口闭塞动脉的实验表明热惯量和血液灌流用激光多普勒成像测量之前和期间血流 8之间的线性关系。集中的bioheat模式也被用来估计在 9静脉闭塞体积描记法的指尖血管反应性。尽管上述努力，量化的皮肤，从皮肤温度测量血液灌流是具有挑战性的，因为缺乏敏感性，皮下脂肪厚度的依赖，和其他促成因素，如血管收缩，血管扩张，和运动伪影10。在这个协议中，我们使用了红外摄像头捕捉到组织温度变化的反应在室温下的热刺激。被选中的热刺激，使血管收缩和血管扩张的影响减少到最低限度。进一步的建模和测量工作是必要的，以划定之间的血管功能指数和皮肤组织灌注的一个可靠的定量关系。 ＃3，我们使用的仿射变换的共同登记任务。然而，鉴于这两个相机都用不同的角度在三维空间中的定位，它是申请有关的两个摄像机之间的3D转换的转型可能更准确。目前，我们正在探索这个方向涉及的相机使用极线几何在三维空间的外在校准。

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		<th>Catalogue Number</th>
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<td>Higgins Calligraphy waterproof black ink</td>
<td>&nbsp;</td>
<td>Sanford</td>
<td>44314</td>
<td>diluted to 1%</td>
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<td>Hamamatsu ORCA ER deep cooling CCD camera</td>
<td>&nbsp;</td>
<td>Hamamatsu, Bridgewater, NJ</td>
<td>C4742-80-12AG</td>
<td>&nbsp;</td>
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<td>Varispec SNIR liquid crystal tunable filter</td>
<td>&nbsp;</td>
<td>Cambridge Research Inc., Cambridge, MA</td>
<td>VIS-10-HC-20</td>
<td>&nbsp;</td>
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<td>ThermoVision A40 infrared camera</td>
<td>&nbsp;</td>
<td>FLIR Systems Inc.,Sweden</td>
<td>A40</td>
<td>&nbsp;</td>
<tr>
<td>Thorlabs OSL1 high intensity fiber light source</td>
<td>&nbsp;</td>
<td>Thorlabs</td>
<td>OSL1</td>
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dual-mode imaging of cutaneous tissue oxygenation and vascular function

相应的检测，分期，和许多健康疾病，如慢性伤口治疗，皮肤组织氧合和血管功能的准确评估是很重要的。我们报告一个非侵入性和非接触的皮肤组织氧合和血管功能成像双模成像系统的发展。成像系统集成的红外摄像机，CCD相机，液晶可调谐滤波器和高强度纤维光源。一个LabVIEW接口设备的控制，同步，图像采集，处理和可视化编程。 CCD相机所拍摄的多光谱图像被用来重建组织氧合图。分别用红外相机拍摄的动态热成像图像重建血管的功能图。皮肤组织氧合和血管功能图像共通过受托标记注册。在人类的双模影像系统的性能特点进行了测试。

Watch this Scientific Journal Video about Dual-mode Imaging of Cutaneous Tissue Oxygenation and Vascular Function at JoVE.com

Dual-mode Imaging of Cutaneous Tissue Oxygenation and Vascular Function

双模式成像系统开发的非接触的皮肤组织氧合和血管功能的评估。

双模式成像，皮肤组织氧合和血管功能

Accurate assessment of cutaneous tissue oxygenation and vascular function is important for appropriate detection, staging, and treatment of many health ...

dual-mode-imaging-cutaneous-tissue-oxygenation-vascular

Research

JoVE Journal

Medicine

16.4K 次观看.  The Ohio State University. 双模式成像系统开发的非接触的皮肤组织氧合和血管功能的评估。

视频: Dual-mode Imaging of Cutaneous Tissue Oxygenation and Vascular Function

Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive surgical procedures. Although this is an effective method of assessing in vitro diaphragm activity, it involves non-survival surgery. The application of non-invasive ultrasound imaging as an in vivo procedure is beneficial since it not only reduces the number of animals sacrificed, but is also suitable for monitoring disease progression in live mice. Thus, our ultrasound imaging method may likely assist in the development of novel therapies that alleviate muscle injury induced by various respiratory diseases. Particularly, in clinical diagnoses of obstructive lung diseases, ultrasound imaging has the potential to be used in conjunction with other standard tests to detect the early onset of diaphragm muscle fatigue. In the current protocol, we describe how to accurately evaluate diaphragm contractility in a mouse model using a diagnostic ultrasound imaging technique.

Diagnostic ultrasound imaging has proven to be effective in diagnosing various respiratory diseases in human and animal subjects. We demonstrate a comprehensive ultrasound protocol utilized by Dr. Zuo's lab to analyze diaphragm kinetics specifically in mouse models. This is also a non-invasive research technique which can provide quantitative information on mouse respiratory muscle function.

小鼠膈肌功能诊断超声成像

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive ...

科研

医学

Assessment of Vascular Function in Patients With Chronic Kidney Disease

Patients with chronic kidney disease (CKD) have significantly increased risk of cardiovascular disease (CVD) compared to the general population, and this is only partially explained by traditional CVD risk factors. Vascular dysfunction is an important non-traditional risk factor, characterized by vascular endothelial dysfunction (most commonly assessed as impaired endothelium-dependent dilation [EDD]) and stiffening of the large elastic arteries. While various techniques exist to assess EDD and large elastic artery stiffness, the most commonly used are brachial artery flow-mediated dilation (FMDBA) and aortic pulse-wave velocity (aPWV), respectively. Both of these noninvasive measures of vascular dysfunction are independent predictors of future cardiovascular events in patients with and without kidney disease. Patients with CKD demonstrate both impaired FMDBA, and increased aPWV. While the exact mechanisms by which vascular dysfunction develops in CKD are incompletely understood, increased oxidative stress and a subsequent reduction in nitric oxide (NO) bioavailability are important contributors. Cellular changes in oxidative stress can be assessed by collecting vascular endothelial cells from the antecubital vein and measuring protein expression of markers of oxidative stress using immunofluorescence. We provide here a discussion of these methods to measure FMDBA, aPWV, and vascular endothelial cell protein expression.

The degree of vascular dysfunction and contributing physiological mechanisms can be assessed in patients with chronic kidney disease by measuring brachial artery flow-mediated dilation, aortic pulse-wave velocity, and vascular endothelial cell protein expression.

血管功能在慢性肾脏疾病的评估

Patients with chronic kidney disease (CKD) have significantly increased risk of cardiovascular disease (CVD) compared to the general population, and this ...

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 ...

Chronic Thromboembolic Pulmonary Hypertension and Assessment of Right Ventricular Function in the Piglet

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. Pulmonary Hypertension (PH) was induced in 3-week-old piglets by a progressive obstruction of the pulmonary vascular bed. A ligation of the left Pulmonary Artery (PA) was performed first through a mini-thoracotomy. Second, weekly embolizations of the right lower pulmonary lobe were done under fluoroscopic guidance with n-butyl-2-cyanoacrylate during 5 weeks. Mean Pulmonary Arterial Pressure (mPAP) measured by ritght heart catheterism, increased progressively, as well as Right Atrial pressure and Pulmonary Vascular Resistances (PVR) after 5 weeks compared to sham animals. Right Ventricular (RV) structural and functional remodeling were assessed by transthoracic echocardiography (RV diameters, RV wall thickness, RV systolic function). RV elastance and RV-pulmonary coupling were assessed by Pressure-Volume Loops (PVL) analysis with conductance method. Histologic study of the lung and the right ventricle were also performed. Molecular analyses on RV fresh tissues could be performed through repeated transcutaneous endomyocardial biopsies. Pulmonary microvascular disease in obstructed and unobstructed territories was studied from lung biopsies using molecular analyses and pathology. Furthermore, the reliability and the reproducibility was associated with a range of PH severity in animals. Most aspects of the human CTEPH disease were reproduced in this model, which allows new perspectives for the understanding of the underlying mechanisms (mitochondria, inflammation) and new therapeutic approaches (targeted, cellular or gene therapies) of the overloaded right ventricle but also pulmonary microvascular disease.

Chronic Thromboembolic Pulmonary Hypertension (CTEPH) and Right Ventricular (RV) dysfunction were induced in piglets by progressive obstruction of the pulmonary arteries. Consequences were remarkably similar to those observed in CTEPH patients. This animal model would be a very useful tool for pathophysiology and therapeutic experiments on CTEPH and RV failure.

慢性血栓栓塞性肺动脉高压和右心室功能的仔猪评估

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. ...

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the silicone compound, via a 26 G catheter, into the portal or hepatic vein. After silicone injection, organs were explanted and prepared for ex-vivo micro-CT (&#181;CT) scanning. The silicone injection procedure is technically challenging. Achieving a successful outcome requires extensive microsurgical experience from the surgeon. One of the challenges of this procedure involves determining the adequate perfusion rate for the silicone compound. The perfusion rate for the silicone compound needs to be defined based on the hemodynamic of the vascular system of interest. Inappropriate perfusion rate can lead to an incomplete perfusion, artificial dilation and rupturing of vascular trees.
The 3D reconstruction of the vascular system was based on CT scans and was achieved using preclinical software such as HepaVision. The quality of the reconstructed vascular tree was directly related to the quality of silicone perfusion. Subsequently computed vascular parameters indicative of vascular growth, such as total vascular volume, were calculated based on the vascular reconstructions. Contrasting the vascular tree with silicone allowed for subsequent histological work-up of the specimen after &#181;CT scanning. The specimen can be subjected to serial sectioning, histological analysis and whole slide scanning, and thereafter to 3D reconstruction of the vascular trees based on histological images. This is the prerequisite for the detection of molecular events and their distribution with respect to the vascular tree. This modified silicone injection procedure can also be used to visualize and reconstruct the vascular systems of other organs. This technique has the potential to be extensively applied to studies concerning vascular anatomy and growth in various animal and disease models.

Tools used for visualizing vascular regeneration require methods for contrasting the vascular trees. This film demonstrated a delicate injection technique used to achieve optimal contrasting of the vascular trees and illustrate the potential benefits resulting from a detailed analysis of the resulting specimen using &#181;CT and histological serial sections.

之后，正常小鼠70％部分肝切除血管和实质再生的可视化

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

主动脉钙化和炎症血管平滑肌细胞和成像的钙化

Cardiovascular disease is the leading cause of morbidity and mortality in the world.  Atherosclerotic plaques, consisting of lipid-laden macrophages and ...

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. ...

Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with 'One Stop Shop' Near-infrared Spectroscopy

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic demand. Reactive hyperemia (in response to a brief period of tissue ischemia) is an independent predictor of cardiovascular events and provides important insight into vascular health and vasodilatory capacity. Skeletal muscle oxidative capacity is equally important in health and disease, as it determines the energy supply for myocellular processes. Here, we describe a simple, non-invasive approach using near-infrared spectroscopy to assess each of these major clinical endpoints (reactive hyperemia, neurovascular coupling, and muscle oxidative capacity) during a single clinic or laboratory visit. Unlike Doppler ultrasound, magnetic resonance images/spectroscopy, or invasive catheter-based flow measurements or muscle biopsies, our approach is less operator-dependent, low-cost, and completely non-invasive. Representative data from our lab taken together with summary data from previously published literature illustrate the utility of each of these end-points. Once this technique is mastered, application to clinical populations will provide important mechanistic insight into exercise intolerance and cardiovascular dysfunction.

Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with &#039;One Stop Shop&#039; Near-infrared Spectroscopy

Here, we describe a simple, non-invasive approach using near-infrared spectroscopy to assess reactive hyperemia, neurovascular coupling and skeletal muscle oxidative capacity in a single clinic or laboratory visit.

"一站式" 近红外光谱对骨骼肌神经血管的耦合、氧化能力及微血管功能的作用

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic ...

Veno-Venous Extracorporeal Membrane Oxygenation in a Mouse

The use of extracorporeal membrane oxygenation (ECMO) has increased substantially in recent years. ECMO has become a reliable and effective therapy for acute as well as end-stage lung diseases. With the increase in clinical demand and prolonged use of ECMO, procedural optimization and prevention of multi-organ damage are of critical importance. The aim of this protocol is to present a detailed technique of veno-venous ECMO in a non-intubated, spontaneously breathing mouse. This protocol demonstrates the technical design of the ECMO and surgical steps. This murine ECMO model will facilitate the study of pathophysiology related to ECMO (e.g., inflammation,bleeding and thromboembolic events). Due to the abundance of genetically modified mice, the molecular mechanisms involved in ECMO-related complications can also be dissected.

Here we present a protocol describing the technique of veno-venous extracorporeal membrane oxygenation (ECMO) in a non-intubated, spontaneously breathing mouse. This murine model of ECMO can be effectively implemented in experimental studies of acute and end-stage lung diseases.

小鼠静脉内膜氧化术

The use of extracorporeal membrane oxygenation (ECMO) has increased substantially in recent years. ECMO has become a reliable and effective therapy for ...

Measurement of Tissue Oxygenation Using Near-Infrared Spectroscopy in Patients Undergoing Hemodialysis

Near-infrared spectroscopy (NIRS) has recently been applied as a tool to measure regional oxygen saturation (rSO2), a marker of tissue oxygenation, in clinical settings including cardiovascular and brain surgery, neonatal monitoring and prehospital medicine. The NIRS monitoring devices are real-time and noninvasive, and have mainly been used for evaluating cerebral oxygenation in critically ill patients during an operation or intensive care. Thus far, the use of NIRS monitoring in patients with chronic kidney disease (CKD) including hemodialysis (HD) has been limited; therefore, we investigated rSO2 values in some organs during HD. We monitored rSO2 values using a NIRS device transmitting near-infrared light at 2 wavelengths of attachment. The HD patients were placed in a supine position, with rSO2 measurement sensors attached to the foreheads, the right hypochondrium and the lower legs to evaluate rSO2 in the brain, liver and lower leg muscles, respectively. NIRS monitoring could be a new approach to clarify changes in organ oxygenation during HD or factors affecting tissue oxygenation in CKD patients. This article describes a protocol to measure tissue oxygenation represented by rSO2 as applied in HD patients.

We present a protocol to measure regional oxygen saturation (rSO2) in hemodialysis (HD) patients by using a near-infrared spectroscopy monitor. The rSO2 value is an index of tissue oxygenation. This noninvasive and real-time monitoring could be useful for confirming changes in organ oxygenation during HD.

使用近红外光谱测量血液透析患者的组织氧合

Near-infrared spectroscopy (NIRS) has recently been applied as a tool to measure regional oxygen saturation (rSO2), a marker of tissue oxygenation, in ...