关于微血管回应空气污染微粒的结果复制与环境与健康展望24权限。经证实，在气象和空气质量数据进行了亲切比利时皇家气象研究所和佛兰芒语环境局提供的。视网膜图像分析软件，从北费里尔博士（工程麦迪逊学院和眼底照相读片中心，眼科及视觉科学系，威斯康星大学麦迪逊分校）获得。 Tijs Louwies和埃利纳教务长与一个VITO奖学金支持。埃利纳教务长认为佛兰德科学基金的上进研究奖学金。蒂姆·S. Nawrot是一个欧洲研究委员会开始拨款持有人。

哈瑟尔特大学的伦理委员会和大学医院安特卫普批准了研究。参加了他们的书面知情同意参加。 1，仪器设置<ol><li>从数字眼底相机和该单元的主块取出黑色保护壳。 </li><li>打开电池仓，将电池放在相机。不要断开连接电池和主机的电线。 </li><li>用螺丝将摄像头到主单元和连接的两根导线。主机连接到电网和计算机附带的USB电缆。 </li><li>通过开关的开/关按钮“上的”启动主单元。通过开关的开/关按钮“上的”启动相机。 </li><li>启动计算机。这将防止在主单元和计算机之间的连接错误。 </li></ol> 2，采集照片<ol><li>启动视网膜意马NG控制软件（并填写所需的密码）。该软件是数码相机视网膜的一部分（见资料表的链接）。 </li><li>通过点击在屏幕的左上部分的“学习”图标展开研究。对于新患者，填写所有的细节，如患者ID，患者姓名，出生日期等，如果患者已经在系统中，填写“病人ID”和使用“搜索历史记录列表”。对患者的名字双击展开研究。 </li><li>让患者在镜头前要坐一坐，把他/她的下巴的下巴休息，对其余的额头和额头上“锁定”头拍摄照片。 </li><li>让患者直视摄像机的镜头。移动摄像机在水平（XY）平面的右眼或左眼。 </li><li>使用颚台来定位，出现在照相机显示器上的两个圆圈内的患者的角膜。微调用车轮上的操纵杆。 </li><li>移动相机向前，向后，侧向和在XY平面上，以定位在圆内的病人的瞳孔。确保学生形成了一个连贯的圆圈。通过这样做，在患者的虹膜将在两片被分割。 </li><li>使用操纵杆上的“背面触发”，从角膜到视网膜的切换。在这个阶段，患者应注意观察了绿灯。让病人看开了绿灯。 </li><li>通过比对两条线，弹出当车轮在操纵杆的基部被接通聚焦照相机。转动车轮，直至两条线形成一条连续的线。 </li><li>使用绿色光源放置在眼睛的照片的最佳位置。如果需要，可使用箭头按钮的右侧照相机的移动灯光。定位在绿色光的方式，视盘为中心的照相机显示器上。 </li><li>搜索2的白色斑点的出现switchi后ng到视网膜（在步骤2.7）。要找到点，移动设备在XY平面上。这些点是作为一个模糊的色斑明显。移动单元向前或向后，直到模糊污点变成明亮的白色斑点。在明亮的圆斑，画面的质量越好。定位点，直到两个是可见的。使用操纵杆上的小轮带来的斑点到相机显示屏的中间。 </li><li>确认两行（步骤2.8），形成一条连续的线。视盘为中心的照相机显示器上，并通过两个明亮的白点侧接。采取了射击上的操纵杆顶部的按钮视网膜照片。 </li><li>通过按电脑屏幕右下角的“研究完成”按钮来保存照片。完成研究会自动保存在地图画面，并关闭研究。 </li></ol>视网膜照片3。分析<ol><li>确定由MEAS规模比例uring的黄斑（黄斑）的中心之间以及视神经盘中心（盲区）的距离。解剖学这个距离被确定为4500微米或2.5倍视盘的直径，而后者是约1800微米。确保该距离测量像素。由黄斑和盲区之间除以4500的距离（以像素为单位）计算的缩放比例。 </li><li>打开视网膜血管分析软件“伊万”。 注：该软件是在美国威斯康星大学麦迪逊分校创建。详细信息伊万的使用是从手册。 </li><li>在规模比例填充，然后继续完成配置。 </li><li>验证三黄圈出现在视网膜照片。缩放比例确定的内圆的半径，并且包围视盘。验证该中间点的内圈是在中间点的视盘。如果不是这种情况，调整圆的，使用光标键位置。中间和外层圆的半径分别为2倍和3倍比内侧圆的半径大，分别为。在这种方式中，区域A和B在从视盘的固定距离被创建。 </li><li>检查视网膜图像中，在照片的中央视盘。这可确保在区域B中的图像的锐聚焦，这将有利于分级处理（ 图4A）。 </li><li>注意，该软件会自动检测血管和分配这些船只为小静脉（ 图4B）。 </li><li>区分的基础上的生理差异小动脉和微静脉之间的血管。动脉将被指示在红色和小静脉中的蓝色（ 图4C）。使用以下准则来确定每艘船： <ol><li>确定容器颜色。动脉有一个打火机橙红色具有强烈的中心光反射。脉有较深的紫色- 红颜色很少或没有中央对光反射消失。 </li><li>确定容器中的过程。动脉往往是直的，更流畅的轮廓;他们都是路径和轮廓更加规整。静脉一般都比较曲折，而在外形和直径更不规则。静脉是更广泛的，直径在盘缘比相应的动脉。 </li><li>通过观察前述容器的注释识别容器。原则上，小动脉相间的小静脉。因此，如果在不同的微静脉被测量，下一容器是更可能是一个小动脉。 </li><li>定义交叉的方式。作为一般规则，小动脉不交叉小动脉和小静脉不交叉的小静脉。如果不明身份的船只穿过内静脉分支或远端B区，则未知船只的动脉。如果它穿过内动脉分支或远端B区，那么它是一个小静脉。 </li><li>通过跟踪他们找出更小的分支proximally的从母体血管分支，它的身份可以是显而易见的，从前两个准则。使用船舶之间的角度来区分通道和支化。 注意：关卡常常几乎垂直（90°），或者，如果两个容器都追猎平行，相交的角可以是非常浅的（小于30°）。支化通常比垂直稍差（与从30°到45°的两个分支之间的角度）。 </li><li>选择在分级区域中的容器的整个长度上。确保所选容器的标准偏差不超过10小于标准偏差的值指示了良好的测量。 </li></ol></li><li>使用软件工具来选择未由软件本身选择的船只。同样的规则也适用于这些船只是由软件自动选择的船只。 </li><li>确定中央视网膜小动脉和小静脉当量（CRAE和CRVE自动伊凡）。 </li><li>使用帕尔和哈伯德28日修订后的公式各自的分支血管的女儿计算CRAE和CRVE。 注：树干和树枝之间的关系，用实验导出的支化系数，给出了下面的两个公式来近似容器当量。 IVAN使用六大小动脉和小静脉计算CRAE和CRVE。该公式应用在迭代过程来配对六大动脉（或静脉）获得中央动脉（或小静脉）血管相当于罚金。 动脉： <img alt="式（1）" fo:content-width="1in" src="/files/ftp_upload/51904/51904eq1.jpg" /> （1） 静脉： <img alt="式（2）" fo:content-width="1in" src="/files/ftp_upload/51904/51904eq2.jpg" /> （2） 其中W 1，W 2和W的窄分支部，该宽分支部，并且父躯干的宽度，分别。 注：假设塔吨视网膜照片六大动脉是120，110，100，90，80，和70微米宽。放120和70代入式（1），以及110和80，和100和90的第一次迭代之后有三个值：122.2，120.0和118.4。通过配对122.2和118.4，得到149.8执行下一次迭代。携带在中间数（120.0）的最后一次迭代。对149.8和120.0，得到168.7的CRAE。 </li></ol>

作者宣称，他们没有实际的或潜在竞争的经济利益。

<html>视网膜图像分析，提出作为一个方便的工具，用于研究在流行病学研究微血管反应。当操作者经验丰富，只需不到5分钟取眼底照片。此外，可用于从幼年到老年参与者这个不显眼的过程可视化的微循环。 文献在增加相对于形态学变化之间的关联，在视网膜脉管系统（例如改变在容器口径，几何图案等）和可修改的生活方式和环境风险因素15,16。实验和流行病学的...

微循环是由血管直径小于150微米，并且包括最小的阻力动脉，小动脉，毛细血管和小静脉。这些血管弥补循环系统的很大一部分并在维持心血管健康具有重要作用。 150微米的血管直径是一种生理和物理限制。容器，直径小于150微米的流变学特性，从大动脉不同。此外，大多数的自动调节的电阻发生变化，从150微米的下游在血管床表现出血流自动调节1。微循环有两个重要的功能。其主要功能是提供细胞与氧和代谢底物以匹配组织需求和排出废物和二氧化碳。在交换血管和微血管血流模式的数目的改变降低了有效交换的表面积，并且可以LEAd，来次优的组织灌注，未能满足代谢需求2。此外，静压力降低的血管床内的微循环在调节整体外周阻力3的作用。 视网膜是层状组织衬眼睛内部。它的主要功能是将入射光转换成被进一步传播到视觉皮层用于处理视觉信息的神经信号。视网膜的功能是看外界和所有参与该过程的眼结构是光学透明的。这使得视网膜组织访问的微血管4的非侵入性的成像。视网膜成像被用来识别眼疾。例如，黄斑变性的一种高级形式可以导致由于异常的血管生长成黄斑视力丧失。这些血管往往更可渗透的，可能会流血克和内或下方的视网膜渗漏血液和蛋白质。后者的事件是负责向光感受器的不可逆损伤。青光眼的相关发展具有破坏性的神经节细胞及其轴突。这一过程的作用导致拔罐视盘，它可以在视网膜图像5可以观察到的。糖尿病视网膜病变是高血糖导致的视网膜血管壁损伤造成的。这可能导致局部缺血，新血管的生长和在血管几何网络的变化。此外，血-视网膜屏障可能会受到破坏，从而引起扩张hyperpermeable毛细血管和动脉瘤6的泄漏。 视网膜微血管显示同源性在心脏，肺和大脑7中发现的微血管床。经确定了影响大脑的微循环全身性疾病可引起视网膜平行的变化。动脉NArrowing和视网膜小动脉增强光的反射与血管发育异常，脑白质病变与腔隙是由脑小血管病变引起的8有关。 à显著关系较窄视网膜静脉，改变的视网膜微血管网络和早老性痴呆症的发生之间被发现。因此建议患者大脑具有改变的脑微血管那也是可观察到的视网膜中的9。 证据也越来越多关于视网膜血管的改变与冠状动脉心脏疾病10,11之间的相关性。视网膜动脉与视网膜静脉（A / V）的直径之间的比例已被证明是一个敏感的代理，以反映高血压和动脉粥样硬化12。变窄的动脉和静脉增宽，导致降低的A / V比，证实了中风和心肌梗死的风险13。高血压可以引起直接的视网膜缺血，视网膜梗塞的变得像棉絮斑和深层视网膜白斑14可见。塞尔和Sasongko最近总结了文献中，他们得出的结论是暴露在生活方式和环境危险因素（ 如饮食，体力活动，吸烟和空气污染），可诱导的视网膜微血管床15的形态变化。重要的是，这样的眼底改变与心血管危险因素有关，甚至在疾病的16个临床表现。 显著增加心血管疾病发病率和死亡率的发生归因于长期和短期暴露于颗粒物质空气污染17,18。研究表明，颗粒物（PM），空气污染的一个重要部分，导致心血管病的发展和诱发心血管事件19,20。功能的损害微血管床认为在观察协会的作用。在这方面，暴露于空气污染和动脉变窄的视网膜之间的关联已经报道由亚达和同事21。视网膜动脉管径较窄和小静脉管径是4607参与者动脉粥样硬化（MESA）是生活在地区增加长期和短期暴露于PM 2.5（颗粒物的多种族研究之间更广泛≤2.5微米直径）21。引起的慢性空气污染暴露全身性炎症可导致较宽的小静脉的直径22。这证实了报道吸烟对视网膜微血管床23的影响研究。短期空气污染暴露与健康成人微血管的变化（22-63岁）与视网膜眼底照相24测量之间的关系最近出版的报告。一个increa本身在PM 10（粒子状物质≤10微米的直径）和BC（黑碳，燃烧副产物可以被用作用于与交通有关的柴油机废气的代理）用在小动脉口径24,25的减少相关联。 在这个科学视频协议，该程序的描述来收集眼睛的眼底图像，进行图像分析，以获得小动脉和小静脉血管的口径，并且计算中央视网膜小动脉当量（CRAE）和中央视网膜小静脉当量（CRVE）。视网膜成像是获得越来越多的关注，因为视网膜，使微血管和图像的一个不显眼的分析，可以从早期收集到老年26,27的唯一组织。 CRAE和CRVE似乎反映改变的生活方式和环境的心血管疾病危险因素对微血管的影响敏感的参数。在手稿中，可重复性容器的分析论证。此外，在流行病学研究视网膜微血管分析的应用如通过总结我们在重复测量设计，获得了专注于微粒空气污染暴露24的影响的研究结果。

<table border="0" cellpadding="0" cellspacing="0" width="970">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:321px;">Name of Material/ Equipment</td>
			<td style="width:223px;">Company</td>
			<td style="width:196px;">Catalog Number</td>
			<td style="width:231px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Canon CR-2 nonmydriatic retinal camera&#160;</td>
			<td>Hospithera (Brussels, Belgium)</td>
			<td></td>
			<td>http://www.usa.canon.com/cusa/healthcare/products/eyecare/digital_non_mydriatic_retinal_cameras/cr_2. Any other retinal camera with comparable resolution and specifications can be used for the analysis of the retinal microvasculature. Compatibility should&#160; be checked before starting a study.</td>
		</tr>
		<tr height="357">
			<td height="357" style="height:357px;width:321px;">IVAN: Vessel Measurement Software</td>
			<td style="width:223px;"></td>
			<td style="width:196px;"></td>
			<td style="width:231px;">This software can be used without charge for scientific purpose. It can be obtained by contacting Dr. Nicola Ferrier (Madison School of Engineering and the Fundus Photograph Reading 
			Center, Department of Ophthalmology and Visual Sciences, University of Wisconsin&#8211;Madison). http://directory.engr.wisc.edu/me/faculty/ferrier_nicola. Phone: (608) 265-8793, 
			Fax: (608) 265-2316 or e-mail: ferrier@engr.wisc.edu 
			&#160;</td>
		</tr>
	</tbody>
</table>

fundus photography as a convenient tool to study microvascular responses to cardiovascular disease risk factors in epidemiological studies

微循环是由血管直径小于150微米。它构成了循环系统的很大一部分并在维持心血管健康具有重要作用。视网膜是一种组织的线眼的内部，这是允许的微血管的非侵入性分析的唯一组织。现在，高品质的眼底图像可以使用数码相机来获取。视网膜图像可以被收集在5分钟或更少，即使没有瞳孔的扩张。这不显眼的和快速的程序的可视化微循环是有吸引力的流行病学研究应用，并从幼年到老年监测心血管的健康。 影响循环全身性疾病可以导致在视网膜脉管渐进的形态学变化。例如，改变在视网膜动脉和静脉的血管口径已患有高血压，动脉粥样相关硬化和中风和心肌梗死的风险增加。容器的宽度是使用图像分析软件和六个最大的动脉的宽度衍生和静脉总结在中央视网膜小动脉当量（CRAE）和中央视网膜小静脉当量（CRVE）。后者的功能已经显示，研究可修改的生活方式和环境心血管疾病的危险因素的影响是有用的。 该程序以获取眼底图像和分析步骤，以获得CRAE和CRVE描述。的CRAE和CRVE重复测量的变异系数是小于2％，并在信度是非常高的。使用面板的研究中，视网膜血管口径在微粒空气污染的短期变化的快速反应，对心脑血管发病率和死亡率的一个已知危险因素，报道。总之，视网膜成像提议作为一种方便的工具的工具流行病学studies到学习微血管应对心血管疾病的危险因素。

的CRAE和CRVE测定重复性 22〜56岁，无临床诊断心血管疾病的61个人的小组被招募研究技术的可重复性和中央视网膜小动脉当量（CRAE）和视网膜中央小静脉当量（CRVE）确定的范围内评价者变异。每个单独的右眼睛眼底是5分钟使用眼底相机（ 图1和2）的时间内拍摄到的两倍。这个过程已完成的连续4天，大约在一天的同一时间。变化在±CRAE的标准偏差和5分钟...

Watch this Scientific Journal Video about Fundus Photography as a Convenient Tool to Study Microvascular Responses to Cardiovascular Disease Risk Factors in Epidemiological Studies at JoVE.com

Fundus Photography as a Convenient Tool to Study Microvascular Responses to Cardiovascular Disease Risk Factors in Epidemiological Studies

视网膜图像分析是一个不显眼的过程可视化微循环。心血管疾病危险因素的影响，可能会导致视网膜血管口径的变化。的程序，以取得眼底图像和步骤，用于计算口径是所述的容器中。

眼底摄影作为一种方便的工具，研究微血管回应流行病学研究心血管疾病的风险因素

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an ...

fundus-photography-as-convenient-tool-to-study-microvascular

Research

JoVE Journal

Medicine

19.0K 次观看.  Flemish Institute for Technological Research (VITO). 视网膜图像分析是一个不显眼的过程可视化微循环。心血管疾病危险因素的影响，可能会导致视网膜血管口径的变化。的程序，以取得眼底图像和步骤，用于计算口径是所述的容器中。 

视频: Fundus Photography as a Convenient Tool to Study Microvascular Responses to Cardiovascular Disease Risk Factors in Epidemiological Studies

Spectral Karyotyping to Study Chromosome Abnormalities in Humans and Mice with Polycystic Kidney Disease

Conventional method to identify and classify individual chromosomes depends on the unique banding pattern of each chromosome in a specific species being analyzed 1, 2. This classical banding technique, however, is not reliable in identifying complex chromosomal aberrations such as those associated with cancer. To overcome the limitations of the banding technique, Spectral Karyotyping (SKY) is introduced to provide much reliable information on chromosome abnormalities.
SKY is a multicolor fluorescence in-situ hybridization (FISH) technique to detect metaphase chromosomes with spectral microscope 3, 4. SKY has been proven to be a valuable tool for the cytogenetic analysis of a broad range of chromosome abnormalities associated with a large number of genetic diseases and malignancies 5, 6. SKY involves the use of multicolor fluorescently-labelled DNA probes prepared from the degenerate oligonucleotide primers by PCR. Thus, every chromosome has a unique spectral color after in-situ hybridization with probes, which are differentially labelled with a mixture of fluorescent dyes (Rhodamine, Texas Red, Cy5, FITC and Cy5.5). The probes used for SKY consist of up to 55 chromosome specific probes 7-10.
The procedure for SKY involves several steps (Figure 1). SKY requires the availability of cells with high mitotic index from normal or diseased tissue or blood. The chromosomes of a single cell from either a freshly isolated primary cell or a cell line are spread on a glass slide. This chromosome spread is labeled with a different combination of fluorescent dyes specific for each chromosome. For probe detection and image acquisition,the spectral imaging system consists of sagnac interferometer and a CCD camera. This allows measurement of the visible light spectrum emitted from the sample and to acquire a spectral image from individual chromosomes. HiSKY, the software used to analyze the results of the captured images, provides an easy identification of chromosome anomalies. The end result is a metaphase and a karyotype classification image, in which each pair of chromosomes has a distinct color (Figure 2). This allows easy identification of chromosome identities and translocations. For more details, please visit Applied Spectral Imaging website (<a href="http://www.spectral-imaging.com/" target="_blank">http://www.spectral-imaging.com/</a>).
SKY was recently used for an identification of chromosome segregation defects and chromosome abnormalities in humans and mice with Autosomal Dominant Polycystic Kidney Disease (ADPKD), a genetic disease characterized by dysfunction in primary cilia 11-13. Using this technique, we demonstrated the presence of abnormal chromosome segregation and chromosomal defects in ADPKD patients and mouse models 14. Further analyses using SKY not only allowed us to identify chromosomal number and identity, but also to accurately detect very complex chromosomal aberrations such as chromosome deletions and translocations (Figure 2).

Spectral Karyotyping (SKY) is an advanced cytogenetics technique to identify genomic and chromosomal aberrations. This technique takes advantage of chromosome painting probes, which allow classification of all chromosomes. SKY can also identify complex chromosome aberrations and segregation defects in mice and humans with various diseases, including polycystic kidney disease.

光谱核型多囊肾病的研究在人类和小鼠的染色体异常

Conventional method to identify and classify individual chromosomes depends on the unique banding pattern of each chromosome in a specific species being ...

科研

医学

Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated blood is the aorta. The aorta may become affected in diseases such as atherosclerosis and aneurysm. Researchers investigating these diseases would benefit from direct observation of the aorta to characterize disease progression as well as to evaluate efficacy of potential therapeutics. The goal of this protocol is to describe proper isolation and excision of the aorta to aid investigators researching cardiovascular disease. Isolation and excision of the aorta allows investigators to look at gross morphometric changes as wells as allowing them to preserve and stain the tissue to look at histologic changes if desired. The aorta may be used for molecular studies to evaluate protein and gene expression to discover targets of interest and mechanisms of action. This technique is superior to imaging modalities as they have inherent limitations in technology and cost. Additionally, primary isolated cells from a freshly isolated and excised aorta can allowing researchers to perform further in situ and in vitro assays. The isolation and excision of the aorta has the limitation of having to sacrifice the animal however, in this case the benefits outweigh the harm as it is the most versatile technique in the study of aortic disease.

Pathology of the aorta can lead to severe morbidity and mortality, therefore research of disease progression and potential therapies is warranted. Here, we present a protocol to isolate and excise the murine aorta to aid researchers in their investigation of cardiovascular disease.

隔离和小鼠主动脉切除;心血管疾病研究的一个多功能的技术

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated ...

Adapting Human Videofluoroscopic Swallow Study Methods to Detect and Characterize Dysphagia in Murine Disease Models

This study adapted human videofluoroscopic swallowing study (VFSS) methods for use with murine disease models for the purpose of facilitating translational dysphagia research. Successful outcomes are dependent upon three critical components: test chambers that permit self-feeding while standing unrestrained in a confined space, recipes that mask the aversive taste/odor of commercially-available oral contrast agents, and a step-by-step test protocol that permits quantification of swallow physiology. Elimination of one or more of these components will have a detrimental impact on the study results. Moreover, the energy level capability of the fluoroscopy system will determine which swallow parameters can be investigated. Most research centers have high energy fluoroscopes designed for use with people and larger animals, which results in exceptionally poor image quality when testing mice and other small rodents. Despite this limitation, we have identified seven VFSS parameters that are consistently quantifiable in mice when using a high energy fluoroscope in combination with the new murine VFSS protocol. We recently obtained a low energy fluoroscopy system with exceptionally high imaging resolution and magnification capabilities that was designed for use with mice and other small rodents. Preliminary work using this new system, in combination with the new murine VFSS protocol, has identified 13 swallow parameters that are consistently quantifiable in mice, which is nearly double the number obtained using conventional (i.e., high energy) fluoroscopes. Identification of additional swallow parameters is expected as we optimize the capabilities of this new system. Results thus far demonstrate the utility of using a low energy fluoroscopy system to detect and quantify subtle changes in swallow physiology that may otherwise be overlooked when using high energy fluoroscopes to investigate murine disease models.

This study successfully adapted human videofluoroscopic swallowing study (VFSS) methods for use with murine disease models for the purpose of facilitating translational dysphagia research.

适应人类Videofluoroscopic燕子研究的方法来检测和检定吞咽困难的小鼠疾病模型

This study adapted human videofluoroscopic swallowing study (VFSS) methods for use with murine disease models for the purpose of facilitating ...

Using Adeno-associated Virus as a Tool to Study Retinal Barriers in Disease

M&#252;ller cells are the principal glial cells of the retina. Their end-feet form the limits of the retina at the outer and inner limiting membranes (ILM), and in conjunction with astrocytes, pericytes and endothelial cells they establish the blood-retinal barrier (BRB). BRB limits material transport between the bloodstream and the retina while the ILM acts as a basement membrane that defines histologically the border between the retina and the vitreous cavity. Labeling M&#252;ller cells is particularly relevant to study the physical state of the retinal barriers, as these cells are an integral part of the BRB and ILM. Both BRB and ILM are frequently altered in retinal disease and are responsible for disease symptoms.
There are several well-established methods to study the integrity of the BRB, such as the Evans blue assay or fluorescein angiography. However these methods do not provide information on the extent of BRB permeability to larger molecules, in nanometer range. Furthermore, they do not provide information on the state of other retinal barriers such as the ILM. To study BRB permeability alongside retinal ILM, we used an AAV based method that provides information on permeability of BRB to larger molecules while indicating the state of the ILM and extracellular matrix proteins in disease states. Two AAV variants are useful for such study: AAV5 and ShH10. AAV5&#160;has a natural tropism for photoreceptors but it cannot get across to the outer retina when administered into the vitreous when the ILM is intact (i.e., in wild-type retinas). ShH10 has a strong tropism towards glial cells and will selectively label M&#252;ller glia in both healthy and diseased retinas. ShH10 provides more efficient gene delivery in retinas where ILM is compromised. These viral tools coupled with immunohistochemistry and blood-DNA analysis shed light onto the state of retinal barriers in disease.

To investigate the blood-retinal barrier permeability and the inner limiting membrane integrity in animal models of retinal disease, we used several adeno-associated virus (AAV) variants as tools to label retinal neurons and glia. Virus mediated reporter gene expression is then used as an indicator of retinal barrier permeability.

使用腺相关病毒作为一种工具来研究疾病中的视网膜障碍

M&#252;ller cells are the principal glial cells of the retina. Their end-feet form the limits of the retina at the outer and inner limiting membranes ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

静脉介入型号：研究旁路移植通畅一个合适的模型

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

Smartphone Fundus Photography

Smartphone fundus photography is a simple technique to obtain ocular fundus pictures using a smartphone camera and a conventional handheld indirect ophthalmoscopy lens. This technique is indispensable when picture documentation of optic nerve, retina, and retinal vessels is necessary but a fundus camera is not available. The main advantage of this technique is the widespread&#160;availability of smartphones that allows documentation of macula and optic nerve changes in many settings that was not previously possible. Following the well-defined steps detailed here, such as proper alignment of the phone camera, handheld lens, and the patient&#39;s pupil, is the key for obtaining a clear retina picture with no interfering light reflections and aberrations. In this paper, the optical principles of indirect ophthalmoscopy and fundus photography will be reviewed first. Then, the step-by-step method to record a good quality retinal image using a smartphone will be explained.

Fundus photography normally requires specialized fundus cameras that are not always available in all clinical settings.&#160;Here, a simple method to record ocular fundus images using a smartphone camera and a conventional high-plus handheld indirect ophthalmoscopy lens is described.

智能手机眼底摄影

Smartphone fundus photography is a simple technique to obtain ocular fundus pictures using a smartphone camera and a conventional handheld indirect ...

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

As a functional status of microcirculation, microvascular vasomotion is important for the delivery of oxygen and nutrients and the removal of carbon dioxide and waste products. The impairment of microvascular vasomotion might be a crucial step in the development of microcirculation-related diseases. In addition, the highly vascularized pancreatic islet is adapted to support endocrine function. In this respect, it seems possible to infer that the functional status of pancreatic islet microvascular vasomotion might affect pancreatic islet function. Analyzing the pathological changes of the functional status of pancreatic islet microvascular vasomotion may be a feasible strategy to determine contributions that pancreatic islet microcirculation makes to related diseases, such as diabetes mellitus, pancreatitis, etc. Therefore, this protocol describes using a laser Doppler blood flow monitor to determine the functional status of pancreatic islet microvascular vasomotion, and to establish parameters (including average blood perfusion, amplitude, frequency, and relative velocity of pancreatic islet microvascular vasomotion) for evaluation of the microcirculatory functional status. In a streptozotocin-induced diabetic mouse model, we observed an impaired functional status of pancreatic islet microvascular vasomotion. In conclusion, this approach for assessing pancreatic islet microvascular vasomotion in vivo may reveal mechanisms relating to pancreatic islet diseases.

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

Pancreatic islet microvascular vasomotion regulates islet blood distribution and maintains the physiological function of islet &#946; cells. This protocol describes using a laser Doppler monitor to determine the functional status of pancreatic islet microvascular vasomotion in vivo and to assess the contributions of pancreatic islet microcirculation to pancreatic-related diseases.

激光多普勒: 一种测量胰岛微血管血管舒缩体内的工具

As a functional status of microcirculation, microvascular vasomotion is important for the delivery of oxygen and nutrients and the removal of carbon ...

Conducting Maximal and Submaximal Endurance Exercise Testing to Measure Physiological and Biological Responses to Acute Exercise in Humans

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic responses to acute exercise are predominantly influenced by the duration and intensity of the exercise regimen. As exercise is increasingly thought of as a therapeutic treatment and/or diagnostic tool, it is important that standardizable methodologies be utilized to understand the variability and to increase the reproducibility of exercise outputs and measurements of responses to such regimens. To that end, we describe two different cycling exercise regimens that yield different physiologic outputs. In a maximal exercise test, exercise intensity is continually increased with a greater workload resulting in an increasing cardiopulmonary and metabolic response (heart rate, stroke volume, ventilation, oxygen consumption and carbon dioxide production). In contrast, during endurance exercise tests, the demand is increased from that at rest, but is raised to a fixed submaximal exercise intensity resulting in a cardiopulmonary and metabolic response that typically plateaus. Along with the protocols, we provide suggestions on measuring physiologic outputs that include, but are not limited to, heart rate, slow and forced vital capacity, gas exchange metrics, and blood pressure to enable the comparison of exercise outputs between studies. Biospecimens can then be sampled to assess cellular, protein, and/or gene expression responses. Overall, this approach can be easily adapted into both short- and long-term effects of two distinct exercise regimens.

To assess the influence of exercise intensity on physiologic and biologic responses, two different exercise testing protocols were utilized. Methods outlining exercise testing on a cycle ergometer as an incremental maximal oxygen consumption test and endurance, steady state submaximal endurance test are described.

进行最大和次极大耐力运动测试, 测量人类急性运动的生理和生物学反应

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic ...

The Dyspepsia Educational Tool As a Novel Aid in Dyspepsia Management

Digital educational tools have a well-established role in current healthcare. In particular, disorders that are managed non-pharmacologically benefit from this development, as it enables patient engagement in self-management. Dyspepsia is a condition thought to arise from gastric and duodenal perturbations, brain-gut axis disturbances, and dietary factors. Behavioral interventions are a major part of dyspepsia treatment, hence patient engagement and motivation through education is essential. Protocols that describe the development process of such educational tools are scarce. We provide a methodology describing development of a dyspepsia educational tool. Assessment of users' needs is the first step, followed by a literature search. The content is developed based on the main themes and entered into a content management system, to build the program. Final adjustments are made after a pilot test of the tool. The presented protocol can be used as a guide for development of a digital dyspepsia educational tool or as a tool for similar situations.

This protocol describes the development process of a digital dyspepsia educational tool. Assessment of unmet needs and literature, content development, and building of the tool are presented. The methodology can be used as a guide for future development of digital educational tools.

消化不良教育工具作为消化不良管理中的一种新帮助

Digital educational tools have a well-established role in current healthcare. In particular, disorders that are managed non-pharmacologically benefit from ...

A Cryoinjury Model to Study Myocardial Infarction in the Mouse

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we demonstrate a murine cryoinjury infarct model that generates precise infarct sizes with high reproducibility and replicability. In brief, after intubation and sternotomy of the animal, the heart is lifted from the thorax. The probe of a handheld liquid nitrogen delivery system is applied onto the myocardial wall to induce cryoinjury. Impaired ventricular function and electrical conduction can be monitored with echocardiography or optical mapping. Transmural myocardial remodeling of the infarcted area is characterized by collagen deposition and loss of cardiomyocytes. Compared to other models (e.g., LAD-ligation), this model utilizes a handheld liquid nitrogen delivery system to generate more uniform infarct sizes.

This article demonstrates a model to study cardiac remodeling after myocardial cryoinjury in mice.

研究小鼠心肌梗死的低温损伤模型

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we ...