作者想，持续的支持和技术援助，以感谢他对这项工作的精辟的审查和威廉Sotka Koby Sheffy，博士。 这项工作是支持的，一部分由来自美国国立卫生研究院的资助（K12 HL087746，RC2HL103400，1U01HL100397），以及加州烟草相关疾病的研究计划的加州大学（18XT - 0098）。

一，准备病人远藤的PAT研究<ol><li>在研究之前，确保病人禁食至少4个小时，已经有至少8个小时，咖啡因，烟草，维生素或药物，可能会影响血管张力忍住。患者不妨使用厕所前的研究。 </li><li>远藤PAT的研究应在一个安静，光线昏暗，温度控制的考场进行，以减少血管张力的波动。 </li><li>手机或传呼设备应保持沉默，而限制性的服装可能干扰血流武器应予删除。病人还应该删除的手表，戒指，或其他首饰，手或手指。 </li><li>检查任何畸形或受伤，可能会影响研究的病人的手指。不要将一个被切断或受伤的手指上的探针。指甲不应该延长超过5mm或超出1 / 5英寸的手指组织的一角。修剪或文件如有必要，以避免损坏内部膜的PAT探头和传感探头的地区取代手指指甲。 </li><li>食指建议为研究对象，但是，如果这个手指是不合适的，不同的数字（拇指除外），可使用，只要使用同一手指双手。 </li><li>患者应为15分钟，仰卧和舒适，从而达到心血管稳态。沿每个病人的两侧放置两手臂的支持者。 </li><li>测量血压使​​用的控制臂（远藤PAT的研究过程中没有遮​​挡手臂。 </li><li>放置在手臂上的血压袖带在远藤的PAT研究闭塞。应用袖带紧贴，但没有多余的压力。不要助长此时的袖口。 </li></ol> II.Prepare内PAT系统的研究<ol><li>启动内PAT 2000软件，单击工具栏上创建一个新的病人档案的“以病人信息”图标。 </li><li>完整的病人信息的对话框，包括病人编号，姓名（可选），年龄，性别，身高，体重，收缩压和舒张压。可选字段允许自由文本的意见。在预先定义的列表中的Patographer名称字段选择你的名字。如果你的名字尚未插入，添加列表和选择。 </li><li>选择两个新的PAT探头和连接的气动电管。要连接的探针，插入探头缝隙连接器“选项卡，并轻轻按压到探头的连接器，直至其卡入到位。 </li><li>放入支持臂插座连接的探针，然后按内PAT 2000设备的顶部的“放气”按钮。 </li></ol> III.Conduct一个远藤的PAT研究<ol><li>患者食指完全放入探针，与患者确认他或她能感觉到非常探头结束，按“膨胀”内PAT 2000设备的顶部按钮。 </li><li>放置在相邻中指基地的泡沫锚环。确保泡沫环和八传感器请勿触摸。否则环机械干扰传感器。 </li><li>创建一个大约7 - 10CM，气动电管循环。循环应延长从PAT传感器和相邻的手指上的泡沫环，而其余的油管EndoPAT设备连接到指出油管手指的尖端。 </li><li>位置病人的手臂，手臂上的支持和手指晃动着自由关闭的边缘的支持，使前臂支持。确保探头不与任何物体接触，包括手臂的支持，泡沫环，管，床垫或其他手指。 </li><li>让患者不要移动手指，因为这将创建机械构件。重要的是要放松整个研究的病人。解释的病人，在测试过程中，你会抬高手臂袖口，并在这段时间里，他们可能会感到有些不适，麻木或刺痛。 </li><li>远藤帕特的计算机接口，点击“待机”图标。时基调整到1分钟，并在屏幕上，以最大限度地提高信号的清晰度调整信号增益。检查从两个探头的PAT信号描记，以确认他们artifactual信号。如果artifactual信号存在，veriify探头不接触的任何东西，该病人是不动的手指。 </li><li>要开始的研究中，单击“转到”计算机界面上的图标。开始计时，点击“开始/停止计时器”图标。这将启动一个五分钟的计数基线录音期间。五分钟后，停止秒表，通过点击“开始/停止计时器”图标。 </li><li>告诉病人，你会在flate闭塞阶段和袖口，他或她应该保持放松，而不是移动手指。 </li><li>迅速膨胀到了病人的收缩压200mmHg 60mmHg以上的超收缩压血压袖带，取其较高，再次启动秒表。手血流完全停止从闭塞手臂PAT信号的情况下验证。为了确认闭塞增加渠道闭塞侧的屏幕上的增益至20,000的对侧端的增益，同时保持不变。减少的渠道，既为30秒的时间。验证，你不遵守任何信号在一个周期相匹配的控制臂的信号，说明这是一个不完全闭塞。如果是这种情况，然后再充气袖带，直到没有信号看到。袖带充气至300mmHg最大。 </li><li>这将启动一个五分钟的计数动脉闭塞录音期间。对闭塞期的结束，告诉病人你要释放的，他们应该继续，以避免自己的手指移动的袖口和的。整整5分钟后，紧缩的袖口突然尽快通过点击“开始/停止计时器”图标，并停止秒表。 </li><li>再次点击“开始/停止计时器”图标，启动五分钟后闭塞，记录周期。五分钟后停止计时器，并点击“停止测试”图标来完成这项研究。该探测器将自动放气。 </li><li>删除的探头，磁带，并从患者的手指泡沫环和断开气动电管的PAT探头。丢弃的探头。 </li></ol> IV.Review和分析<ol><li>负载的研究文件在屏幕上使用的负载图标。要运行的自动分析，按一下“魔术师坚持”图标。闭塞期间将突出显示为蓝色，和测试结果将显示，包括反应性充血指数（RHI）和心率（HR），在屏幕的右侧栏，。 </li><li>审查的其他数据，包括研究的参数，计算变量，病人信息，和信号质量的措施，请点击图标“打开最后计算结果”。这将打开一个试算表，研究参数和执行所有的分析结果与包含从最近期的分析的数据表的最后一行。 </li></ol>五，代表性的成果一个代表远藤PAT个人内皮血管扩张功能正常进行的一项研究的屏幕如图1所示。个人内皮血管扩张功能不全的远藤的PAT研究的一个代表性的画面如图2所示。 <img src="/files/ftp_upload/2167/2167fig1.jpg" alt="图1" /> 图1：内皮血管舒张功能正常。代表记录的个体与正常内皮血管扩张功能，特点是袖口释放后相对于基线信号的幅度增加。 <img src="/files/ftp_upload/2167/2167fig2.jpg" alt="图2" /> 图2：内皮血管舒张功能障碍。内皮血管扩张功能不全的个人代表录音。

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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+Vascular+Function+in+Obese+Adolescents+with+Insulin+Resistance.">Impaired Vascular Function in Obese Adolescents with Insulin Resistance.</a> J Pediatr. 155 (5), 678-682 (2009).</li><li>Hirata, Y., Nagata, D., Suzuki, E., Nishimatsu, H., Suzuki, J., Nagai, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+and+treatment+of+endothelial+dysfunction+in+cardiovascular+disease.">Diagnosis and treatment of endothelial dysfunction in cardiovascular disease.</a> Int Heart J. 51 (1), 1-6 (2010).</li><li>Hamburg, N. M., Keyes, M. J., Larson, M. G., Vasan, R. S., Schnabel, R., Pryde, M. M., Mitchell, G. F., Sheffy, J., Vita, J. A., Benjamin, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cross-Sectional+Relations+of+Digital+Vascular+Function+to+Cardiovascular+Risk+Factors+in+the+Framingham+Heart+Study.">Cross-Sectional Relations of Digital Vascular Function to Cardiovascular Risk Factors in the Framingham Heart Study.</a> Circulation. 117 (19), 2467-2474 (2008).</li><li>Truschel, E., Jarczok, M. N., Fischer, J. E., Terris, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+ambulatory+assessment+of+digital+reactive+hyperemia:+Concurrent+validity+with+known+cardiovascular+risk+factors+and+potential+confounding.">High-throughput ambulatory assessment of digital reactive hyperemia: Concurrent validity with known cardiovascular risk factors and potential confounding.</a> Prev Med. 49 (6), 468-4672 (2009).</li><li>Rubinshtein, R., Kuvin, J. T., Soffler, M., Lennon, R. J., Lavi, S., Nelson, R. E., Pumper, G. M., Lerman, L. O., Lerman, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+Endothelial+Function+by+Non-invasive+Peripheral+Arterial+Tonometry+Predicts+Late+Cardiovascular+Adverse+Events.">Assessment of Endothelial Function by Non-invasive Peripheral Arterial Tonometry Predicts Late Cardiovascular Adverse Events.</a> Eur Heart. , (2010).</li><li>Yamaoka-Tojo, M., Tojo, T., Kosugi, R., Hatakeyama, Y., Yoshida, Y., Machida, Y., Aoyama, N., Masuda, T., Izumi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+ezetimibe+add-on+therapy+for+high-risk+patients+with+dyslipidemia.">Effects of ezetimibe add-on therapy for high-risk patients with dyslipidemia.</a> Lipids Health Dis. 8 (1), 41-41 (2009).</li></ol>

支持视频开发伊塔马尔医药有限公司博士库克是NOS途径，从他收到版税的诊断和治疗应用的斯坦福大学拥有的专利的发明者。

全世界的发病率和死亡率的主要原因是动脉粥样硬化性血管疾病，导致中风，心肌梗塞，心脏衰竭，肾功能不全，动脉瘤破裂出血或栓塞，间歇性跛行和坏疽1,2。内皮功能障碍是导致这3个动脉粥样硬化疾患的病理生理过程的最早事件之一。另外，血管内皮功能障碍，有助于疾病的进展，促进炎症和血栓形成。4-6。传统的心血管危险因素与血管内皮血管扩张功能不全 7-11 。内皮血管扩张功能障碍，可以检测到在看似健康的个人发展心血管疾病的12风险。此外，内皮血管扩张功能障碍的调查结果，主要不良心血管事件的预测，以及死亡率13-16。 由内皮细胞释放的血管扩张剂包括如前列环素，如肾上腺髓质素和心钠素肽，小分子，如内皮依赖性超极化因子，一氧化碳和一氧化氮的 17等血管扩张的前列腺素。此外，在血管内皮细胞产生的极化电流的影响可以传输到底层的小型船只的血管平滑肌放松其中18。可以得到一个间接产生这些内皮细胞的血管扩张的影响评估研究血管反应性。最常见的方法来评估内皮调节血管反应性的非侵入性已多普勒超声检测 19肱动脉血流介导的血管舒张。 流体流动的牵引力刺激血管内皮细胞释放血管扩张，最突出的一氧化氮20。在前臂血流增加，在反应性充血 19诱发的肱动脉超声可以观察到这种现象。这项技术已被广泛利用文件内皮血管扩张功能障碍与心血管病危险因素的关联;内皮血管扩张功能障碍的各种生物标志物的关系，如C -反应肽，或不对称二甲基精氨酸（一氧化氮合酶的内源性拮抗剂），;内皮血管扩张功能与营养和生活方式的修改以及使用血管紧张素转换酶抑制剂，血管紧张素受体 ​​阻滞剂，他汀类药物，胰岛素增敏剂或阿司匹林 21-23校正。 的血流介导的血管舒张的肱动脉超声评估需要昂贵的设备，是运营商高度依赖，响应有一个非常小的动态范围和信号信噪比低。新的方法来解决这些问题，内皮血管扩张功能的评估都需要 24。远藤PAT的2000是一个快速的非侵入性的方式，以评估内皮血管扩张功能的一种新的方法。该技术提供的一个反应性充血指数（RHI），使内皮血管扩张功能的指示的计算值。 RHI是后前在闭塞的手臂，在对照组相同的比例相对闭塞的PAT信号比，并为基线血管张力纠正。 EndoPAT使用的研究表明，奥镁得分反映NO的生物利用度25。奥镁在26冠状动脉内皮血管扩张功能的测量和肱口蹄疫 27相关。更大程度的心血管疾病患者表现出分数较低的28和价值观也与血管内皮功能受损及心血管疾病的29-33风险相关的其他条件。值得注意的是，奥镁值出现35心血管结局的预测。一个低的奥镁（血管内皮功能障碍）是可以逆转与治疗 36 。 总之，我们已经演示了如何执行一个可靠和可重复性的测试与远藤的PAT 2000内皮血管扩张功能。测试是无创，易于操作，是一个有用的研究工具。其在血管内皮功能的临床监测和剪裁疾病管理的效用正在调查中。

assessing endothelial vasodilator function with the endo-pat 2000

内皮是一个微妙的细胞单层，行所有的血管，并包括全身和毛细淋巴管。凭借的盔甲，它分泌旁分泌的因素，血管内皮细胞调节底层的血管平滑肌收缩和增殖状态，以及与血液循环元素的血管壁的相互作用。由于其在调解船只语调和增长，其作为网关循环免疫细胞的地位，以及其止血和凝血的地方性法规的核心作用，正常运转的血管内皮是心血管健康的关键。相反，在大多数血管疾病的最早的障碍是血管内皮功能障碍。 在动脉循环，健康内皮细胞一般对对血管平滑肌的血管扩张的影响。这里有许多方法来评估内皮的血管扩张功能。远藤PAT的2000是一个新的设备，用于在一个快速，非侵入性的方式来评估内皮的血管扩张功能。与常用的双面超超声技术评估血流介导的血管舒张，它是完全不取决于运营商和设备是为了幅度不太昂贵。波形的PAT（外周动脉音）信号测量用新型改性体积描记探头对位于每只手的食指，被称为内皮细胞介导的​​设备记录的数字脉冲的变化。在PAT信号的内皮细胞介导的​​变化，引起下游的充血反应。充血是诱发咬合通过5分钟，一方面利用充气袖带肱动脉的血流量。反应性充血反应是由系统自动计算。使用后和前闭塞值创建一个PAT的比例。这些值是标准化的测量，从对侧的手臂，作为控制非内皮依赖性的系统性影响。最值得注意的是，这种正常化控制交感神经流出的波动，可能诱发叠加的充血反应的变化，在周围动脉音。 在这个视频中，我们将演示如何使用远藤的PAT 2000执行内皮血管扩张功能的临床相关的评估。

Watch this Scientific Journal Video about Assessing Endothelial Vasodilator Function with the Endo-PAT 2000 at JoVE.com

Assessing Endothelial Vasodilator Function with the Endo-PAT 2000

表现出一种非侵入性的程序，以评估血管内皮功能是使用远藤的PAT 2000。

评估与远藤的PAT 2000内皮血管舒张功能

The endothelium is a delicate monolayer of cells that lines all blood vessels, and which comprises the systemic and lymphatic capillaries. By virtue of ...

assessing-endothelial-vasodilator-function-with-the-endo-pat-2000

Research

JoVE Journal

Medicine

43.4K 次观看.  Stanford University. 表现出一种非侵入性的程序，以评估血管内皮功能是使用远藤的PAT 2000。

视频: Assessing Endothelial Vasodilator Function with the Endo-PAT 2000

Dual-mode Imaging of Cutaneous Tissue Oxygenation and Vascular Function

Accurate assessment of cutaneous tissue oxygenation and vascular function is important for appropriate detection, staging, and treatment of many health disorders such as chronic wounds. We report the development of a dual-mode imaging system for non-invasive and non-contact imaging of cutaneous tissue oxygenation and vascular function. The imaging system integrated an infrared camera, a CCD camera, a liquid crystal tunable filter and a high intensity fiber light source. A Labview interface was programmed for equipment control, synchronization, image acquisition, processing, and visualization. Multispectral images captured by the CCD camera were used to reconstruct the tissue oxygenation map. Dynamic thermographic images captured by the infrared camera were used to reconstruct the vascular function map. Cutaneous tissue oxygenation and vascular function images were co-registered through fiduciary markers. The performance characteristics of the dual-mode image system were tested in humans.

A dual-mode imaging system was developed for non-contact assessment of cutaneous tissue oxygenation and vascular function.

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 ...

科研

医学

In Vivo Canine Muscle Function Assay

We describe a minimally-invasive and reproducible method to measure canine pelvic limb muscle strength and muscle response to repeated eccentric 
contractions. The pelvic limb of an anesthetized dog is immobilized in a stereotactic frame to align the tibia at a right angle to the femur. Adhesive wrap affixes the 
paw to a pedal mounted on the shaft of a servomotor to measure torque. Percutaneous nerve stimulation activates pelvic limb muscles of the paw to either push (extend) or 
pull (flex) against the pedal to generate isometric torque. Percutaneous tibial nerve stimulation activates tibiotarsal extensor muscles. Repeated eccentric (lengthening) 
contractions are induced in the tibiotarsal flexor muscles by percutaneous peroneal nerve stimulation. The eccentric protocol consists of an initial isometric contraction 
followed by a forced stretch imposed by the servomotor. The rotation effectively lengthens the muscle while it contracts, e.g., an eccentric contraction. During 
stimulation flexor muscles are subjected to an 800 msec isometric and 200 msec eccentric contraction. This procedure is repeated every 5 sec. To avoid fatigue, 4 min rest 
follows every 10 contractions with a total of 30 contractions performed.

In Vivo Canine Muscle Function Assay

We describe a minimally-invasive and painless method to measure canine hindlimb muscle strength and muscle response to repeated eccentric contractions.

在犬体内的肌肉功能测定

We describe a minimally-invasive and reproducible method to measure canine pelvic limb muscle strength and muscle response to repeated eccentric 
...

Assessing Replication and Beta Cell Function in Adenovirally-transduced Isolated Rodent Islets

Glucose homeostasis is primarily controlled by the endocrine hormones insulin and glucagon, secreted from the pancreatic beta and alpha cells, respectively. Functional beta cell mass is determined by the anatomical beta cell mass as well as the ability of the beta cells to respond to a nutrient load. A loss of functional beta cell mass is central to both major forms of diabetes 1-3. Whereas the declining functional beta cell mass results from an autoimmune attack in type 1 diabetes, in type 2 diabetes, this decrement develops from both an inability of beta cells to secrete insulin appropriately and the destruction of beta cells from a cadre of mechanisms. Thus, efforts to restore functional beta cell mass are paramount to the better treatment of and potential cures for diabetes.
Efforts are underway to identify molecular pathways that can be exploited to stimulate the replication and enhance the function of beta cells. Ideally, therapeutic targets would improve both beta cell growth and function. Perhaps more important though is to identify whether a strategy that stimulates beta cell growth comes at the cost of impairing beta cell function (such as with some oncogenes) and vice versa.
By systematically suppressing or overexpressing the expression of target genes in isolated rat islets, one can identify potential therapeutic targets for increasing functional beta cell mass 4-6. Adenoviral vectors can be employed to efficiently overexpress or knockdown proteins in isolated rat islets 4,7-15. Here, we present a method to manipulate gene expression utilizing adenoviral transduction and assess islet replication and beta cell function in isolated rat islets (Figure 1). This method has been used previously to identify novel targets that modulate beta cell replication or function 5,6,8,9,16,17.

This protocol allows one to identify factors that modulate functional beta cell mass to find potential therapeutic targets for the treatment of diabetes. The protocol consists of a streamlined method to assess islet replication and beta cell function in isolated rat islets following manipulation of gene expression with adenoviruses.

复制和β细胞功能评估Adenovirally转分离的啮齿动物胰岛

Glucose homeostasis is primarily controlled by the endocrine hormones insulin and glucagon, secreted from the pancreatic beta and alpha cells, ...

Non-invasive Assessment of Microvascular and Endothelial Function

The authors have utilized capillaroscopy and forearm blood flow techniques to investigate the role of microvascular dysfunction in pathogenesis of cardiovascular disease. Capillaroscopy is a non-invasive, relatively inexpensive methodology for directly visualizing the microcirculation. Percent capillary recruitment is assessed by dividing the increase in capillary density induced by postocclusive reactive hyperemia (postocclusive reactive hyperemia capillary density minus baseline capillary density), by the maximal capillary density (observed during passive venous occlusion). Percent perfused capillaries represents the proportion of all capillaries present that are perfused (functionally active), and is calculated by dividing postocclusive reactive hyperemia capillary density by the maximal capillary density. Both percent capillary recruitment and percent perfused capillaries reflect the number of functional capillaries. The forearm blood flow (FBF) technique provides accepted non-invasive measures of endothelial function: The ratio FBFmax/FBFbase is computed as an estimate of vasodilation, by dividing the mean of the four FBFmax values by the mean of the four FBFbase values. Forearm vascular resistance at maximal vasodilation (FVRmax) is calculated as the mean arterial pressure (MAP) divided by FBFmax. Both the capillaroscopy and forearm techniques are readily acceptable to patients and can be learned quickly.
The microvascular and endothelial function measures obtained using the methodologies described in this paper may have future utility in clinical patient cardiovascular risk-reduction strategies. As we have published reports demonstrating that microvascular and endothelial dysfunction are found in initial stages of hypertension including prehypertension, microvascular and endothelial function measures may eventually aid in early identification, risk-stratification and prevention of end-stage vascular pathology, with its potentially fatal consequences.

Capillaroscopy is a non-invasive, relatively inexpensive methodology for directly visualizing the microcirculation. The forearm blood flow technique provides accepted non-invasive measures of endothelial function.

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 ...

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 ...

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 ...

Isolation of Human Lymphatic Endothelial Cells by Multi-parameter Fluorescence-activated Cell Sorting

Lymphatic system disorders such as primary lymphedema, lymphatic malformations and lymphatic tumors are rare conditions that cause significant morbidity but little is known about their biology. Isolating highly pure human lymphatic endothelial cells (LECs) from diseased and healthy tissue would facilitate studies of the lymphatic endothelium at genetic, molecular and cellular levels. It is anticipated that these investigations may reveal targets for new therapies that may change the clinical management of these conditions. A protocol describing the isolation of human foreskin LECs and lymphatic malformation lymphatic endothelial cells (LM LECs) is presented. To obtain a single cell suspension tissue was minced and enzymatically treated using dispase II and collagenase II. The resulting single cell suspension was then labelled with antibodies to cluster of differentiation (CD) markers CD34, CD31, Vascular Endothelial Growth Factor-3 (VEGFR-3) and PODOPLANIN. Stained viable cells were sorted on a fluorescently activated cell sorter (FACS) to separate the CD34LowCD31PosVEGFR-3PosPODOPLANINPos LM LEC population from other endothelial and non-endothelial cells. The sorted LM LECs were cultured and expanded on fibronectin-coated flasks for further experimental use.

The goal of this protocol is to isolate lymphatic endothelial cells lining human lymphatic malformation cyst-like vessels and foreskins using fluorescence-activated cell sorting (FACS). Subsequent cell culturing and expansion of these cells permits a new level of experimental sophistication for genetic, proteomic, functional and cell differentiation studies.

人淋巴管内皮细胞中的多参​​数荧光激活细胞分选分离

Lymphatic system disorders such as primary lymphedema, lymphatic malformations and lymphatic tumors are rare conditions that cause significant morbidity ...

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including Parkinson's and Huntington's can disrupt it. People with Parkinson's disease, for example, tend to have increased saccadic latencies. Consequently, the quantitative measurement of saccadic eye movements has received considerable attention as a potential biomarker for neurodegenerative conditions. A lot more can be learned about the brain in both health and disease by observing what happens to eye movements when the function of specific brain areas is perturbed. Deep brain stimulation is a surgical intervention used for the management of a range of neurological conditions including Parkinson's disease, in which stimulating electrodes are placed in specific brain areas including several sites in the basal ganglia. Eye movement measurements can then be made with the stimulator systems both off and on and the results compared. With suitable experimental design, this approach can be used to study the pathophysiology of the disease being treated, the mechanism by which DBS exerts it beneficial effects, and even aspects of normal neurophysiology.

This paper describes the use of quantitative measurement of eye movements in conjunction with stimulation of focal areas of the deep brain in order to study physiology, pathophysiology, and the mechanisms of deep brain stimulation.

使用Saccadometry与深部脑刺激来研究正常和病理脑功能

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including ...

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 ...

Nerve-sparing Mid-urethral Obstruction (NeMO) in Female Small Rodents

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal model to investigate bladder remodeling in pBOO are female rodents undergoing partial obstruction at the proximal urethra. Variability in the degree of obstruction and animal mortality are major concerns with proximal obstruction. Furthermore, dissecting around the proximal urethra and bladder neck jeopardizes bladder innervation.
We developed a nerve-sparing mid-urethral obstruction (NeMO) model for pBOO avoiding the disadvantages of the traditional model. We approached the urethra just inferior to the pubic symphysis, which obviated the need for laparotomy as well as for dissection in this area; also, the striated urethral sphincter remained untouched. We performed NeMO in female Sprague-Dawley rats (12 obstructions, 6 sham animals) as well as in female C57/bl6 mice (20 obstructions, 18 sham animals). After two weeks, we evaluated bladder function, bladder mass, and body mass.
We had no mortalities among obstructed- or sham-operated female rats; as described for the traditional proximal pBOO-method, we tied the suture around the proximal urethra and a temporarily placed 0.9 mm metal rod. NeMO induced an 85% increase in bladder mass after two weeks, average residual urine volume was 0.4 mL in partially obstructed rats while only 0.03 mL in sham animals. In mice, we tested 3 sizes of cannulas that we placed along the urethra when tying the suture. We found that using a 27-gauge cannula resulted in over 50% animal mortality; placing the 25-gauge cannula did not yield the desired response in increasing bladder mass; utilizing a 26-gauge cannula yielded favorable results with minimal animal mortality (1/8) yet a significant 2-fold increase in bladder mass.

Nerve-sparing Mid-urethral Obstruction NeMO in Female Small Rodents

Traditional modeling of partial bladder outlet obstruction in rodents is fraught with animal mortality. A denervation injury from dissection around the proximal urethra and bladder neck is also of major concern. We developed and evaluated a safe and reliable mid-urethral obstruction model, avoiding the shortcomings of the traditional model.

在女小鼠害保留神经的中期尿道梗阻（NEMO）

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal ...