这项研究是由大学医学中心的乌得勒支，血管连接BV和EuroTransBio，项目ELANA锁眼（ETB110014）的支持。 Medistim提供的VeriQÇ控制台，以及超声和流量探针计费降低价格。我们承认伊夫林Velema，Marlijn扬森，乔伊斯维瑟，格雷斯克罗夫特，面包车的Martijn Nieuwburg，塞斯Verlaan， 里克·贝克Mansvelt，桑德面包车Thoor，安德烈面包车迪伦的建设性贡献， &#160;从乌得勒支大学中心动物设施的同事。

注:该动物接受人文关怀符合"指南实验动物的护理和使用"编写的实验动物资源研究所，国家研究理事会。乌得勒支大学的动物实验委员会批准的协议。 1.冠状动脉吻合程序注:请始终使用激光防护眼镜当激光激活。 <ol style="margin-left: 40px;"><li>安装三位一体剪辑<ol><li>开的连接器3（ 图1）与动脉瘤施夹器（ 图4A）的上叉，并插入到灌注移植物的管腔，定向远侧（ 图5A）。确保铲叉的全长intraluminally定位。随后，释放涂药。 </li><li>具有无创伤钢夹和carefu近端堵塞乳内动脉LLY冲洗移植"与管腔肝素盐水。接着，介绍激光导管4（ 图2）血管内，通过移植物的远端自由端，插入连接器，以及由外固定夹（ 图3）固定它。 </li><li>通过导管上的接枝引发真空抽吸（通过导管施加真空，真空管连接到导管和一个真空泵），激活激光和激光打孔接枝一个VUE，导致0.8的吻合口2.0毫米。激光加工后直接释放真空吸力。目视检查动脉壁被完全切除。 </li><li>除去接枝（ 即 "瓣"），它被连接到该导管的真空通道的激光打孔片段，并留下迷恋在接枝和连接器（ 图5B）的导管。 </li></ol></li><li>嫁接的非阻塞性连接到录音ipient <ol><li>打开与VASCO涂抹体（ 图4B）的连接器的下叉，并插入到灌注冠状动脉管腔，定向远侧（ 图5C）。确保铲叉的全长intraluminally定位。随后，释放涂药。 </li></ol></li><li>激光打孔动脉切开术<ol><li>启动真空抽吸到冠状壁并随后激光打孔的冠状壁下充满天然冠状动脉血流，导致0.8的孔口2.0毫米。施工期间不要用力到导管。 </li><li>除去固定夹与动脉瘤施夹器，并随后缩回导管。检查，如果激光打孔瓣附连到导管的真空通道并停止真空抽吸（ 图5D）。 </li><li>以下襟翼检索，通过临时剪辑（ 例如 ，钢夹）阻塞接枝的前端。如果一个有趣虚构旁路被确认（ 即，成功的襟翼检索和足够旁路流动），永久结扎用血管夹（ 图5E）的移植物的远端。 </li><li>在案件瓣检索功能衰竭（ 即不完全光刻瓣仍部分附着在冠状动脉壁），收回连接，关闭部分光刻冠状动脉缝合（8-0普理灵），并创建一个新的吻合，首发在步骤1.1.2。 </li></ol></li></ol> 2.动物，麻醉和安乐死<ol style="margin-left: 40px;"><li>动物和麻醉<ol><li>用女性（ 例如，荷兰长白）猪（70-90公斤[公斤]），并喂正常饮食。 </li><li>管理乙酰水杨酸和75毫克的氯吡格雷320毫克（mg）口服，每日开始手术前3天。继续这种抗凝协议直至终止。 </li><li>管理芬太尼25微克（微克），透皮给药3天:25微克24小时的术前和25微克，在对术后镇痛1天术后。 </li><li>麻醉诱导管理氯胺酮（10毫克/千克），咪达唑仑（0.5毫克/公斤），与阿托品（0.04毫克/千克）肌注。 </li><li>随后，通过静脉内线路管理硫喷妥钠（4毫克/千克），咪达唑仑（0.5毫克/千克），舒芬太尼柠檬酸盐（6微克/公斤），和1000/100毫克的阿莫西林 - 克拉维酸（抗生素预防）。 </li><li>插管和通风与氧和空气的混合物（1:1体积/体积）。 </li><li>使用眼药，并随后关闭了眼睛，防止眼睛的脱水。 </li><li>然后施用作为连续静脉内输注咪达唑仑（0.7毫克/千克/小时），舒芬太尼柠檬酸盐（6微克/ kg /小时），泮库溴铵（0.1毫克/千克/小时），和食盐水（300毫升/小时）。首先通过静脉注射线300毫克胺碘酮在500毫升的羟乙基淀粉溶液。 </li><li>插入动脉线股动脉˚F或动脉血压监测和动脉血液样本。 注:在此之前的压力线是功能不手术将启动。一个可检测的疼痛，以防动物不能充分麻醉，通过增加心脏速率和血压。如果检测到增加心脏速率和血压，增加咪达唑仑和枸橼酸舒芬太尼的管理。 </li><li>管理静脉注射美托洛尔（范围5-20毫克），以减轻心脏的机械烦躁不安，直到大约50-70次一个心脏率/分钟时获得的。 </li><li>在手术过程中插入导管经由尿道膀胱。 </li></ol></li><li>恢复<ol><li>停止麻醉如果漏极（参见步骤3.9-3.11）被除去。 </li><li>如果动物是警觉和足够的恢复意识的呼吸拔管后充分，回归动物的笼子（ 即动物住房面积）。不要让动物在无人值守前面这一点。 </li><li>通过使用一个安全的设置将在鼻子前面的0.5升（L），氧气面罩。 </li><li>不返回已经经历手术治疗对其他动物的公司，直到完全恢复的动物。 </li><li>肌内每日口服，每天两次，在第1天作为一种预防性抗生素，美洛昔康（0.4毫克/千克）在第1天和第2术后镇痛;术后，辖synolux（250毫克/20公斤阿莫西林 - 克拉维酸）。 </li></ol></li><li>安乐死<ol><li>充分heparinize由25000 IU的肝素（获得至少4倍的控制值的活化凝血时间[ACT]），以防止验尸凝结。 </li><li>安乐死与戊巴比妥钠（200毫克/千克）静脉注射的动物，开始与尸检前确认死亡 - 该协议的第6部分。 </li></ol></li></ol> 3.手术注:一个标准的手术室是requi红色的程序，包含所有的标准的材料和设备（至少一个血压监视器，心电图装置和脉搏血氧仪）。标准开胸套，同比胸廓内动脉（ITA）拉钩，一显微组和实验专用仪器必须准备和消毒。推荐使用的外科回路和外科头灯。 <ol style="margin-left: 40px;"><li>通过胸骨打开胸腔。直言不讳地剖析（用手指）胸骨的心包。然后看到或分割（用锤子和刻刀）从xyphoid过程长达胸骨柄胸骨。使用bonewax以防止泄漏从胸骨骨髓。 </li><li>收获的左（或右）从第二肋ITA到隔膜，部分heparinize（ACT至少2.5倍的控制值），和剪辑和解剖ITA在远端站点。 </li><li>固定并通过组织稳定在预定的部位呈现目标冠状动脉（外径[OD] 1.6-1.9毫米，用卡尺测量;或者，ID 1.4-1.6毫米，与心外膜超声测定值[ECUS]）。解剖目标冠心病，删除松围外膜组织，并覆盖冠状动脉与罂粟碱浸泡过的纱布目标。 </li><li>制备天然冠状动脉±2.0-3.0厘米的永久结扎近侧吻合（参见第3.7节）由冠状动脉的广泛的横向清扫，以这样的方式一个金属夹钳夹可以完全结扎冠状动脉。 </li><li>解剖ITA的目标区域，±2.0-3.0厘米近侧到远侧自由端，除去关节周围外膜组织，并测量口径（外径2.0-4.0毫米）。 </li><li>构造以如先前描述的，或可替代地三位一体剪辑的吻合术，构建一个手工缝合吻合术，并使用一个分流，以减少心肌缺血。 </li><li>与3中hemoclips近端结扎冠状动脉。确保无副作用分支闭塞和结扎是100％的闭塞性，防止竞争的流动。 注:调整，并通过将临时无创伤结扎夹在冠状动脉，同时维持一个代表平均动脉血压和生理位置心脏的设定接枝流动;创建一个低流量旁路，结扎冠状动脉充分向远侧，并且允许更多的接枝流，结扎冠状动脉相对更近侧10。 </li><li>盖上心包补片吻合，以防止不受控制的牵引到吻合，关闭胸部后。 </li><li>放置纵隔和/或胸膜漏，并连接到抽吸系统。 </li><li>关闭胸部。 </li><li>一旦水渠停止生产，取下排水管。 </li></ol> 4.术考试<ol style="margin-left: 40px;"><li>一般术数据<ol><li>记录ITA和冠状动脉用卡尺（OD）或ECUS（ID的目标尺寸，见第3节。3），吻合口施工时间（分钟或秒），任何吻合口瘘（分类: 如 ，直接止血，渗出，或轻快泄漏5,6），并注意是否需要额外的针以止血。 </li></ol></li><li>运输时间流量测量（TTFM） <ol><li>记录平均流量，流量曲线，舒张期充盈百分比，并且搏动指数（PI），并结合了平均动脉压。 注:现代TTFM控制台自动计算这些变量。 </li><li>放置在远端接枝用含水凝胶的镂空段的传输时间流量探针，以提高探测器的接触。使用不同的探针的大小，以避免移植物的变形或压缩。 </li><li>通过适当的全身血压测量，胸部关闭前才和释放组织稳定，与心脏的生理位置之后，又一次。 </li><li>计算PI由（最大流量最小流量）/平均流量。该PI是indicat吻合11,12的质量或。 </li></ol></li><li>可选:峰值充血流响应<ol><li>在90毫米汞柱的平均动脉压，夹持接 ​​枝为30秒，并随后测量峰值充血潮流响应，±所述组织稳定器6的释放后30分钟。 </li><li>计算出的平均峰值流速移植的平均基线流量在90毫米汞柱分为冠状动脉峰值流速充血反应（ 即吻合口血流储备）。 </li><li> 10分钟后重复测量。 </li></ol></li><li>可选:心外膜超声<ol><li>放置的ECU探针上用含水凝胶的吻合，以提高探测器的接触。收购ITA-LAD吻合的组织稳定稳定了心脏的横向和纵向的形象。 </li><li>记录用的ECU系统吻合的宽度，长度和高度，并评估吻合的几何形状的质量和冠状动脉流出吨RACT 13。如果缩小（ 例如 ，通过背景或侧壁捕获），修改吻合。 注:金属（ 例如，金属钛夹或吻合口连接器）影响成像质量。 </li></ol></li><li>可选:术冠状动脉造影<ol><li>可视化旁路通过一个标准的冠状动脉造影。通过髂动脉引入导管。等级根据菲茨吉标准通畅。 </li></ol></li></ol> 5.跟踪检查<ol style="margin-left: 40px;"><li>冠状动脉造影<ol><li>可视化旁路通过一个标准的冠状动脉造影和档次根据菲茨吉本标准的通畅。 </li></ol></li><li>可选:运输时间流量测量<ol><li>使腋下切口背腹侧线，下面的肋骨弯曲。如果需要的话，部分地去除所述第二或第三肋，并解剖近端ITA。 </li><li>测量并记录移植流过境时间流量测量（见4.2节）。 </li></ol></li><li>可选:血流储备分数与冠状动脉血流储备<ol><li>辖冠状动脉内硝酸甘油（200微克），以防止痉挛。 </li><li>同时测量冠状动脉内压力和流速。记录的压力和流量，并结合主动脉压和ECG信号。 </li><li>计算从（连续3）直接测量远端（冠状动脉）和近端（LITA）向吻合的血流储备分数（FFR），并且在回旋冠状动脉（CX;控制冠状动脉）。进行测量，在基线和期间最大充血，诱发冠状动脉内腺苷（60微克）的推注。 </li><li>计算冠状动脉血流储备（CFR）的最大充血流速由流速比基线14。 </li></ol></li><li>可选:光学相干断层扫描<ol><li>使用频域光学相干来断层扫描（OCT）系统进行成像旁路用自动拉回速度为20mm /秒和对比度通过手动注射连续平齐​​。 </li><li>记录内膜增生和旁路的尺寸（ 即，基准腔区域中的冠状动脉和ITA1.0厘米向下和上游的吻合术，分别和吻合口）7,15。 </li></ol></li></ol> 6.尸检<ol style="margin-left: 40px;"><li>外植，固定和宏观检验<ol><li>为了尽量减少损害旁路的风险，外植体的心脏整块 ，包括胸骨和肋骨，并标记ITA的福尔马林灌注非常近的一部分。 注:一个开放的胸部手术后几天，心脏被完全附着于胸骨结缔组织粘连。执行胸骨切开术可能损坏旁路，并且，因此，不推荐使用。 </li><li>进行灌注，fixati对以修复其生理形状绕行，从而为后续正确的组织学解释。注入的ITA用福尔马林（4％）中的流动在机壳±90毫米汞柱: </li><li>放置一个瓶子用福尔马林（1L）±1米高于心脏。 </li><li>连接的管（ 例如，一个标准输注系统或类似的硅管）瓶子和近侧ITA之间。 </li><li>注入福尔马林进入心脏，经由ITA和吻合，约60分钟或直到所有福尔马林被完全注入。 </li><li>然后，小心地切除用刀片，剪刀和镊子旁路。留下一些血纤维蛋白/瘢痕组织，心肌，近端ITA和冠状动脉，和远端的冠状动脉，附着到吻合。 </li><li>在4％的福尔马林固定的吻合，该ITA（±1厘米上游吻合）的基准部分，和在LAD（±1厘米下游吻合）的基准部分，过夜。 </li><li>打开冠状动脉longitudinally的下壁，并检查使用10或20倍的放大倍率吻合。由垂直拍摄孔口与它旁边的标尺记录吻合口的宽度和长度。接着，测量吻合的宽度和长度的数字。 </li></ol></li><li>组织学分析<ol><li>嵌入吻合和基准部件中的塑料（甲基丙烯酸甲酯）。 </li><li>在横向（或纵向）面用金刚石锯段，开始在下游5mm时，持续达5毫米吻合染色用苏木精和曙红的上游，和。 </li><li>记录并评估血管壁对合，吻合口区，血栓形成，血管内膜增生，血液暴露nonintimal表面16（BENIS;连接器表面的腔内照射和激光边缘[ 即，无论是移植物和冠状动脉的内侧和外膜表面动脉]），急性和慢性炎性细胞反应（多形核细胞，巨噬细胞和异物巨细胞），和组织损伤6。 </li><li>执行使用软件包的测量。 </li></ol></li><li>可选:扫描电子显微镜<ol><li>固定的吻合术，上述的灌注 - 固定后（参见6.1.2），在2％的戊二醛溶液中缓冲的0.1M纯化磷酸盐缓冲液中。 </li><li>放入1％的缓冲四氧化锇吻合1小时即可完成固定。 </li><li>固定后，脱水的吻合在乙醇分级系列（50，70，90，和100％），并在液态CO 2通过使用临界点的方法。 </li><li>接着，打开冠状动脉的后壁和ITA的上壁在吻合部位用锋利的外科刀片。 </li><li>固定在扫描桶的检体，并与一个薄的铂层通过溅射处理覆盖，以提高图像质量。 </li><li>然后计算血管吻合surfaCE（ 即，内皮细胞和/或血小板覆盖的评估）通过使用扫描型电子显微镜6。 </li></ol></li></ol>

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	<li>Halkos, M. E., Puskas, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Off-pump+coronary+surgery:+where+do+we+stand+in+2010.">Off-pump coronary surgery: where do we stand in 2010.</a> Curr Opin Cardiol. 25, 583-588 (2010).</li><li>Lapierre, H., Chan, V., Sohmer, B., Mesana, T. G., Ruel, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimally+invasive+coronary+artery+bypass+grafting+via+a+small+thoracotomy+versus+off-pump:+a+case-matched+study.">Minimally invasive coronary artery bypass grafting via a small thoracotomy versus off-pump: a case-matched study.</a> Eur J Cardiothorac Surg. 40, 804-810 (2011).</li><li>Tulleken, C. A. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+vessel+connectors+and+methods+for+blood+vessel+connection.">Blood vessel connectors and methods for blood vessel connection.</a> US patent. , (2013).</li><li>Van Thoor, A. C. E., Stecher, D., Keizer, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catheter+apparatus+and+method.">Catheter apparatus and method.</a> US patent. , (1995).</li><li>Stecher, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+nonocclusive+laser-assisted+coronary+anastomotic+connector+in+a+rabbit+model.">A new nonocclusive laser-assisted coronary anastomotic connector in a rabbit model.</a> J Thorac Cardiovasc Surg. 145, 1124-1129 (2013).</li><li>Stecher, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+nonocclusive+laser-assisted+coronary+anastomotic+connector+in+an+off-pump+porcine+bypass+model.">The nonocclusive laser-assisted coronary anastomotic connector in an off-pump porcine bypass model.</a> J Thorac Cardiovasc Surg. 147, 1390-1397 (2014).</li><li>Stecher, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Six-Month+Healing+of+the+Nonocclusive+Coronary+Anastomotic+Connector+in+an+Off-Pump+Porcine+Bypass+Model.">Six-Month Healing of the Nonocclusive Coronary Anastomotic Connector in an Off-Pump Porcine Bypass Model.</a> Innovations (Phila). 9, 130-136 (2014).</li><li>Tulleken, C. A., Verdaasdonk, R. M., Berendsen, W., Mali, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+the+excimer+laser+in+high-flow+bypass+surgery+of+the+brain).">Use of the excimer laser in high-flow bypass surgery of the brain).</a> J Neurosurg. 78, 477-480 (1993).</li><li>Doormaal, T. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patency,+flow,+endothelialization+of+the+sutureless+Excimer+Laser+Assisted+Non-occlusive+Anastomosis+(ELANA)+technique+in+a+pig+model.">Patency, flow, endothelialization of the sutureless Excimer Laser Assisted Non-occlusive Anastomosis (ELANA) technique in a pig model.</a> J Neurosurg. 115, 1221-1230 (2011).</li><li>Buijsrogge, M. P., Grundeman, P. F., Verlaan, C. W., Borst, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unconventional+vessel+wall+apposition+in+off-pump+porcine+coronary+artery+bypass+grafting:+low+versus+high+graft+flow.">Unconventional vessel wall apposition in off-pump porcine coronary artery bypass grafting: low versus high graft flow.</a> J Thorac Cardiovasc Surg. 123, 341-347 (2002).</li><li>Walpoth, B. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transit-time+flow+measurement+for+detection+of+early+graft+failure+during+myocardial+revascularization.">Transit-time flow measurement for detection of early graft failure during myocardial revascularization.</a> Ann Thrac Surg. 66, 1097-1100 (1998).</li><li>Ancona, G., Karamanoukian, H. L., Salerno, T. A., Schmid, S., Bergsland, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow+measurement+in+coronary+surgery.">Flow measurement in coronary surgery.</a> Heart Surg Forum. 2, 121-124 (1999).</li><li>Lovstakken, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+flow+imaging:+a+new+two-dimensional+ultrasound+modality+for+enhanced+intraoperative+visualization+of+blood+flow+patterns+in+coronary+anastomoses.">Blood flow imaging: a new two-dimensional ultrasound modality for enhanced intraoperative visualization of blood flow patterns in coronary anastomoses.</a> J Am Soc Echocardiogr. 21, 969-975 (2008).</li><li>Koudstaal, S., et al. <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+coronary+microvascular+resistance+in+the+chronic+infarcted+pig+heart.">Assessment of coronary microvascular resistance in the chronic infarcted pig heart.</a> J Cell Mol Med. 17, 1128-1135 (2013).</li><li>Agostoni, P., Stella, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography:+new+(near-infrared)+light+on+stent+implantation.">Optical coherence tomography: new (near-infrared) light on stent implantation.</a> Heart. 95, 1895-1896 (2009).</li><li>Scheltes, J. S., van Andel, C. J., Pistecky, P. V., Borst, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coronary+anastomotic+devices:+blood-exposed+non-intimal+surface+and+coronary+wall+stress.">Coronary anastomotic devices: blood-exposed non-intimal surface and coronary wall stress.</a> J Thorac Cardiovasc Surg. 126, 191-199 (2003).</li><li>Balkhy, H. H., Wann, L. S., Arnsdorf, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+patency+evaluation+of+new+distal+anastomotic+device+in+internal+mammary+artery+grafts+using+computed+tomography+angiography.">Early patency evaluation of new distal anastomotic device in internal mammary artery grafts using computed tomography angiography.</a> Innovations. 5, 109-113 (2010).</li><li>Matschke, K. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Cardica+C-Port+System:+clinical+and+angiographic+evaluation+of+a+new+device+for+automated,+compliant+distal+anastomoses+in+coronary+artery+bypass+grafting+surgery--a+multicenter+prospective+clinical+trial.">The Cardica C-Port System: clinical and angiographic evaluation of a new device for automated, compliant distal anastomoses in coronary artery bypass grafting surgery--a multicenter prospective clinical trial.</a> J Thorac Cardiovasc Surg. 130, 1645-1652 (2005).</li><li>Suyker, W. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stapled+coronary+anastomosis+with+minimal+intraluminal+artifact:+The+S2+Anastomotic+System+in+the+off-pump+porcine+model.">Stapled coronary anastomosis with minimal intraluminal artifact: The S2 Anastomotic System in the off-pump porcine model.</a> J Thorac Cardiovasc Surg. 127, 498-503 (2004).</li><li>Carrel, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+and+angiographic+results+after+mechanical+connection+for+distal+anastomosis+in+coronary+surgery.">Clinical and angiographic results after mechanical connection for distal anastomosis in coronary surgery.</a> J Thorac Cardiovasc Surg. 127, 1632-1640 (2004).</li><li>Filsoufi, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+distal+coronary+bypass+with+a+novel+magnetic+coupler+(MVP+system).">Automated distal coronary bypass with a novel magnetic coupler (MVP system).</a> J Thorac Cardiovasc Surg. 127, 185-192 (2004).</li><li>Fischell, T. A., Virmani, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracoronary+brachytherapy+in+the+porcine+model:+a+different+animal.">Intracoronary brachytherapy in the porcine model: a different animal.</a> Circulation. 104, 2388-2390 (2001).</li><li>Kostering, H., Mast, W. P., Kaethner, T., Nebendahl, K., Holtz, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+coagulation+studies+in+domestic+pigs+(Hanover+breed)+and+minipigs+(Goettingen+breed).">Blood coagulation studies in domestic pigs (Hanover breed) and minipigs (Goettingen breed).</a> Lab Anim. 17, 346-349 (1983).</li><li>Ancona, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Graft+revision+after+transit+time+flow+measurement+in+off-pump+coronary+artery+bypass+grafting.">Graft revision after transit time flow measurement in off-pump coronary artery bypass grafting.</a> Eur J Cardiothorac Surg. 17, 287-293 (2000).</li><li>Di Giammarco, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Predictive+value+of+intraoperative+transit-time+flow+measurement+for+short-term+graft+patency+in+coronary+surgery.">Predictive value of intraoperative transit-time flow measurement for short-term graft patency in coronary surgery.</a> J Thorac Cardiovasc Surg. 132, 468-474 (2006).</li><li>Kieser, T. M., Rose, S., Kowalewski, R., Belenkie, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transit-time+flow+predicts+outcomes+in+coronary+artery+bypass+graft+patients:+a+series+of+1000+consecutive+arterial+grafts.">Transit-time flow predicts outcomes in coronary artery bypass graft patients: a series of 1000 consecutive arterial grafts.</a> Eur J Cardiothorac Surg. 38, 155-162 (2010).</li><li>Haaverstad, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraoperative+color+Doppler+ultrasound+assessment+of+LIMA-to-LAD+anastomoses+in+off-pump+coronary+artery+bypass+grafting.">Intraoperative color Doppler ultrasound assessment of LIMA-to-LAD anastomoses in off-pump coronary artery bypass grafting.</a> Ann Thorac Surg. 74, 1390-1394 (2002).</li><li>Klein, P., Meijer, R., Eikelaar, J. H., Grundeman, P. F., Borst, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epicardial+ultrasound+in+off-pump+coronary+artery+bypass+grafting:+potential+aid+in+intraoperative+coronary+diagnostics.">Epicardial ultrasound in off-pump coronary artery bypass grafting: potential aid in intraoperative coronary diagnostics.</a> Ann Thorac Surg. 73, 809-812 (2002).</li><li>Dessing, T. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Geometry+assessment+of+coronary+artery+anastomoses+with+construction+errors+by+epicardial+ultrasound.">Geometry assessment of coronary artery anastomoses with construction errors by epicardial ultrasound.</a> Eur J Cardiothorac Surg. 26, 257-261 (2004).</li><li>Budde, R. P., Meijer, R., Dessing, T. C., Borst, C., Grundeman, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+construction+errors+in+ex+vivo+coronary+artery+anastomoses+by+13-MHz+epicardial+ultrasonography.">Detection of construction errors in ex vivo coronary artery anastomoses by 13-MHz epicardial ultrasonography.</a> J Thorac Cardiovasc Surg. 129, 1078-1083 (2005).</li><li>Di Giammarco, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraoperative+graft+verification+in+coronary+surgery:+increased+diagnostic+accuracy+adding+high-resolution+epicardial+ultrasonography+to+transit-time+flow+measurement.">Intraoperative graft verification in coronary surgery: increased diagnostic accuracy adding high-resolution epicardial ultrasonography to transit-time flow measurement.</a> Eur J Cardiothorac Surg. , (2013).</li><li>Pijls, N. H. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coronaire+fysiologie+en+myocardischemie.">Coronaire fysiologie en myocardischemie.</a> Cardiologie. 2, 169-170 (2008).</li></ol>

The authors declare that they have no competing financial interests. C.A.F. Tulleken holds shares in AMJ b.v., Utrecht, The Netherlands. D. Stecher, G. Bronkers, J.O.T. Noest, C.A.F. Tulleken, and M.P. Buijsrogge are registered as co-inventors of patents regarding the ELANA technique, without financial benefits.

本文介绍了一种新的冠状吻合连接器，三位一体夹，以及如何在猪不停跳搭桥模型来评估这种新设备。提出了不同的技术来评估吻合的质量，通过新的连接器或传统构造的便利:术中，术后，以及验尸技术。以及愈合和重塑过程 - - 所述推动吻合的质量和安全性的评价在短期和长期的是最重要的前向冠状动​​脉吻合连接器的未来临床应用。 目前，只有1冠状动脉吻合连接器被应用于临床17,18，多个其他设备表现出不利的实验或临床结果，或者开发商未能向市场推出该产品19-21。相比其他方法促使冠状动脉吻合，三位一体夹包括几个有兴趣的朋友刺痛功能。首先，由于血管壁的非闭塞性连接，冠状动脉调理（ 例如，挖到或分流）是多余的，从而使吻合结构中不流血字段无时间限制，因此，降低了操作的冠状动脉。第二，该结构也相对简单和直接的，既不是一个单独的切口插入冠状动脉，也不将额外的线圈以止血是必要的。第三，连接器是一种低轮廓装置，无需大体积设备部署系统;因此，它不会妨碍旁通结构上难以到达或心脏的偏远地区，以及，因此，它会潜在地通过微创方法延长的可能性血运重建。 关于促进吻合的生物学行为的重要问题尚未得到答复。什么是激光打孔动脉切开成两个ITA与LAD的影响？可以在血液暴露的非内膜表面（ 即，叉的材料和内侧和外膜激光轮圈），相对于小尺寸靶冠状动脉，是通过过量的内膜增生形成在长期的潜在限制？为了回答这些问题，临床前研究中，使用在本文中所描述的猪模型，将评估长期通畅，此外，愈合以及有关内膜增生的形成与后续潜力吻合狭窄重塑的效果。此外，在该临床前研究中，通畅，愈合，和重塑的促进吻合将与对照相比，常规手工缝合，吻合术。猪模型适用于这些人类类似的生理和心脏和冠状动脉的解剖研究问题，因为它的，它的畅通愈合反应（ 如内膜增生形成），其中6个月随访杜尔ATION在猪模型相当于1.5-3年的随访支架人类冠状动脉22。然而，年轻的，健康的猪的动脉不是患病的和兼容的，并且因此不同的心胸外科实践中遇到的人的患病的血管。因此，之前的临床介绍，在连接器的可行性和安全性也将评估在人动脉粥样硬化尸体模型。此外，高凝状态的倾向在猪23被找到。因此，评估对小口径冠状动脉便利地吻合，猪模型是相当具有挑战性。这一点，在此协议中所述抗血小板治疗（75毫克氯吡格雷和320毫克乙酰基水杨酸）的理由。此外，该抗血小板方案是在预期吻合（BENIS）的血液暴露nonintimal表面。在我们以前的研究中，我们发现，前身科伦的吻合nonintimal面元连接后10天完全6,7内皮。对于抗血小板治疗在诊所中的作用，使用这种连接器，应根据内皮的速率。一旦nonintimal表面被内皮化，抗血小板治疗方案可能会被降低。 准分子激光器是一种接触式激光，并会只成功激光打孔血管壁的情况下有充分直接圆周激光 - 组织接触。在吻合结构中最关键的步骤，因此，这是激光导管到移植物的血管壁和冠状动脉的正确位置。这一步对体外尸体模型进行训练（ 如猪心脏），以尽量减少学习曲线。可能出现的情况，这将导致在激光器的瓣检索失败，并且应考虑到一点，这里的描述:1）冠状动脉吻合连接器被设计为接枝连接到冠状动脉，和secondly，以用作激光平台。该连接器呈现出容器壁（ 即，直链组织表面，没有凸块），并允许导管到血管壁中的垂直位置。如果接头MAL-定位（ 如，不完全插入，背景或侧壁捕捉，肌壁间或-adventitial定位），血管壁介绍是次优的（ 即没有直表面）。因此，在故障位置的情况下，每个人都应该重新定位连接器之前的不归路（ 即动脉切开术）的地步。 2）固定夹设计为垂直保持激光导管插入连接器的施工过程中。然而，固定夹不旨在抵抗许多禁忌力，因此，外科医生具有在施工过程中，以支持激光导管。如果没有足够的支持，导管可以脱臼。 3）为了确保最佳的激光 - 组织接触，冠状动脉壁具有被解剖自由ö˚F松围外膜组织，对侧壁。可以肯定的是，激光表面仅由冠状动脉壁，从内膜至其外膜，也没有围外膜组织捕获到连接器。 如果不幸吻合构造失败，应该收回剪辑（通过打开下叉只），然后关闭冠状动脉病变（±2毫米长），具有修复缝合（8-0普理灵）。猪通常是缺血性的压力相当敏感。因此，缺血预处理被阻断静脉修​​复缺陷之前推荐的。接着，新的吻合可以构造远端到第一目标。接枝仍安装在连接器的上叉。所以导管重新定位和固定后，连接器可以直接插入到冠状动脉。 最重要的吻合评估技术是冠状动脉造影（临床金sTANDARD）和组织学（实验金标准，与冠状动脉造影结合）。然而，吻合过境时间流量测量（TTFM）术中质量评估是非常翔实。 TTFM是快速的，非侵入性的，实时的，而且容易，此外，正确的解释可以减少技术，不可见的，错误11,12,24-26的数目。现代TTFM游戏机自动计算并显示实时平均流量，流量曲线，以及搏动指数（PI），以及大量的其他参数。在PI是由（最大流分钟流量）计算/平均流量和是吻合的质量的指标，而在它自己的平均流量是不可靠指标。低的平均流量（&lt;15毫升/分钟）具有良好的PI（&lt;5）和良好的舒张流曲线可以通过在一个小目标冠状动脉用适度的径流完美吻合发现，而良好的平均流量（ "15毫升/分钟）与异常舒张图案和高PI（&gt; 5）则提示吻合缺陷或移植失败（ 即扭转，压缩或扭结移植物）的。在这种情况下，应该考虑修改吻合。因此，吻合的质量的良好评估应包括流动曲线的解释，该搏动指数，平均流量，结合临床状态。然而，所报道的特异性和TTFM的灵敏度并不统一，并且，因此，诊断的准确性是争论。另外，PI的截止值是临床经验，而不是临床研究的基础上凭经验确定。该TTFM控制台，我们目前正处于临床前动物研究使用处置心外膜超声成像。如果仍然关于吻合的流量测量后的质量的不确定性，实时心外膜超声图像可以是在吻合进一步评估有很大的帮助，在此增加了诊断一个ccuracy 27-31。 实验替代TTF测量是峰值充血潮流响应32， 即，冠状动脉血流储备，这是峰值充血流量之比，以下30秒的接枝闭塞，和基部的流动。峰值流量充血反应应&gt; 4的远端吻合。如果吻合是针对近端上的冠状动脉，峰值充血潮流响应可以是略微降低并且应&gt; 3 6。缺席的充血反应流程暗示的吻合技术错误或移植失败。在这种情况下，咨询TTF测量和临床状况，并考虑修改吻合。请注意，与动脉压（因此，总是测量在相同的平均动脉压，一式两份），并且缺血预处理可以峰值充血潮流响应产生负面影响的绝对血流储备而变化。此外，该峰hypere麦克风潮流响应是不是一个有效的方法和绝对截止值尚未确定。我们经验选择了我们的实验经验的基础上截止。 最后，在此协议中描述的吻合术是，其目的和潜在的实验吻合技术在临床微创设置要应用。目前，在本文中给出的技术的应用材料尚未最终确定和市场准备的产品，而是原型工具。仍有改善（ 例如 ，通用的施放和灵活的激光导管）的一个窗口，这将在不久填入。这种新技术具有有趣的潜力，并且将被彻底在临床前研究中使用该协议进行评价。

<html>非体外循环冠状动脉搭桥（OPCAB）手术可能会减少在冠状动脉搭桥手术使用体外循环相关的疾病（ 例如，血栓栓塞并发症，保留过多的液体，输血，并激活免疫系统）并能有利于患者的高风险体外循环和主动脉操作1相关的并发症。微创冠状动脉搭桥手术（ 例如，胸腔镜或机器人辅助外科手术），降低了切口的大小，因此，降低了患者的恢复时间，住院时间，和发病率2。尽管对有需要的病人进行冠状动脉血管重建的潜在益处（的一个子集），通过这些技术尚未普及。其中一个原因是，一个非体外循环微创冠状动脉搭桥手术侵入性的方法在技术上是非常具有挑战性的。 ontent"&gt;为了简化和促进心脏跳动（ 即非体外循环），微创冠状动脉搭桥手术，一个新的冠状吻合连接器开发:三位一体夹3,4的基础上，准分子激光辅助非闭塞吻合（ELANA ）技术5-9。所述的连接器实现了简化，缝合，并接枝到冠状动脉非闭塞性连接，以及受激准分子激光器导管激光打孔吻合的开口，因此，由于完整的非闭塞吻合结构，冠状动脉调理（ 即，阻断或挖到和分流）是没有必要的，而相比之下，传统吻合术，因此，简化了旁路过程。 之前关于前身原型ELANA冠状动脉连接器的研究，展示了其在相对 ​​大动脉的可行性（内径[ID]2.4毫米[毫米]）在急性兔模型5。更多以上，在猪开放胸骨OPCAB模型，适当的愈合具有最小内膜增生被发现在长期6,7。 近日，冠状动脉吻合技术，进一步改善对临床应用。在连接器和受激准分子激光导管的设计修改使一个简化和加速结构（ 即，移植物缝合安装）上，以临床相关，小口径冠状动脉（ID 1.4-1.6毫米）。之前在冠状动脉旁路移植术的临床应用，这种新颖的连接器的安全性和质量将在长期（6个月的随访）猪打开胸骨OPCAB模型进行评估，根据在本文中描述的协议。 这个协议描述了我们的实验猪OPCAB模型，并提供了冠状动脉吻合过程的详细描述。此外，选项为术，postoperati描述已经和吻合，这是极为重要的评估吻合质量的验尸评估。在本文中，代表结果部分总结了猪OPCAB模式（N = 3头猪，与后续5小时），这是之前的临床前研究中进行了试点研究的结果。

<table border="0" cellpadding="0" cellspacing="0" width="1333">
	<tbody>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;"><a name="RANGE!A1:D23">Name of Material/ Equipment</a></td>
			<td style="width:415px;">Company</td>
			<td style="width:139px;">Catalog Number</td>
			<td style="width:267px;">Comments/Description</td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">Trinity Clip</td>
			<td>Vascular Connect b.v., Utrecht, The Netherlands</td>
			<td style="width:139px;"></td>
			<td>Initial Prototype and Proprietary Design</td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">Excimer Laser System CVX-300</td>
			<td>Spectranetics Corp., Colorado Springs, CO</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">Oval laser catheter</td>
			<td>Vascular Connect b.v., Utrecht, The Netherlands</td>
			<td style="width:139px;"></td>
			<td>Initial Prototype and Proprietary Design</td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">Silicon Extension Tube (vacuum tube)</td>
			<td>Medela, Baar, Switzerland</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">Medela Dominant 50 Pump (vacuum pump)</td>
			<td>Medela, Baar, Switzerland</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">Fixation clip</td>
			<td>Vascular Connect b.v., Utrecht, The Netherlands</td>
			<td style="width:139px;"></td>
			<td>Initial Prototype and Proprietary Design</td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">Standard Aneurysm clip applier</td>
			<td>Peter Lazic, GmbH, Tuttlingen, Germany</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">VasCo applicator</td>
			<td>Vascular Connect b.v., Utrecht, The Netherlands</td>
			<td style="width:139px;"></td>
			<td>Initial Prototype and Proprietary Design</td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">Microvascular Acland clamp B-3V</td>
			<td style="width:415px;">S&#38;T Marketing Ltd, Neuhausen,Switzerland</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">Aneurysm clip Yasargil-type, curved, 9 mm</td>
			<td style="width:415px;">Scanlan International, Inc, Saint Paul, Minn</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">Weck Hemoclip</td>
			<td>Teleflex Medical, Research Triangle Park, NC</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">Hemochron Signature Elite</td>
			<td style="width:415px;">International Technidyne Corporation (ITC), Edison, NJ, USA</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">Hemochron Jr. Activated Clotting Time Plus (ACT+) (cartridge)</td>
			<td style="width:415px;">International Technidyne Corporation (ITC), Edison, NJ, USA</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">Arteriotomy shunt</td>
			<td>Medtronic, Inc, Minneapolis, Minn</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">Octopus Evolution AS (cardiac tissue stabilizer)</td>
			<td>Medtronic, Inc, Minneapolis, Minn</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">VeriQ C (TTFM and epicardial ultrasound)</td>
			<td>Medi-Stim ASA, Oslo, Norway</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">Allura Xper FD20</td>
			<td>Philips, Eindhoven, the Netherlands</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">Combowire&#160;</td>
			<td>Volcano Corporation, San Diego, CA, USA</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;">ComboMap system&#160;</td>
			<td>Volcano Corporation, San Diego, CA, USA</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">C7 Dragonfly (frequency domain optical coherence tomography (OCT) system)</td>
			<td>LightLab Imaging, Inc., Westford, MA</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">AnalySiS (software package)</td>
			<td>Soft-Imaging Software GmbH, M&#252;nster, Germany</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:515px;">Philips XL30LAB (scanning electron microscope)</td>
			<td style="width:415px;">FEI Europe, Eindhoven, The Netherlands</td>
			<td style="width:139px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

evaluation of a novel laser-assisted coronary anastomotic connector - the trinity clip - in a porcine off-pump bypass model

为了简化和促进心脏跳动（ 即非体外循环），微创冠状动脉搭桥手术，一个新的冠状吻合连接器，三位一体夹，是基于准分子激光辅助非闭塞吻合技术开发。三位一体夹子接头实现了简化，缝合，并接枝到冠状动脉非闭塞性连接，以及受激准分子激光器的激光导管冲头吻合的开口。因此，由于完整的非闭塞吻合结构，冠状动脉调理（ 例如，闭塞或分流）是没有必要的，而相比之下，传统的吻合术，因此，简化了断开泵的旁路过程。此前在冠状动脉旁路移植术的临床应用，这种新型连接器的安全性和质量将在长期的实验猪非体外循环冠状动脉搭桥（OPCAB）的研究进行评估。在本文中，我们将介绍如何评价吨他在使用各种技术来评估其质量的猪OPCAB模型冠状动脉吻合。代表性的结果总结并目视证实。

我们进行了试点研究之前，新的三位一体剪辑的评价在大型长期的临床前安全性研究，以评估的可行性。在这个试验性研究，3 LITA至LAD吻合（N =每动物1）构建由1研究者（DS）的连接器，在猪OPCAB模型。 5小时随访计划。 冠状动脉吻合连接器启用完全非闭塞，缝合，和快速吻合建设（平均3.4±0.4分钟）。在所有吻合彻底止血证明了100％瓣检索率。手术数据，在表1中列出的，表明在猪模型OPCAB冠状动脉吻合连接器的可行性。以最小的收缩峰，低于5一个PI和一个主要的舒张充盈接枝（舒张期充盈[DF] 80％）在随访被始终如一测量，如在图6中看到的，这是正常的SUG吸引力流量曲线充血性专利冠状动脉移植。平均峰值充血流响应，以下30秒移植闭塞，为5.6±0.5，表明充足的冠状动脉血流储备。以5小时跟进，宏观检验证明专利不吻合腔内血栓形成如可在图8A可以看出， 图7显示出血管造影5周的一个例子跟进，并验宏观和组织学的例子检验结果显示在图8B和C，均清楚地表明一个改造和完全吻合专利5周随访（临床前研究的初步成果）。此外，先前的研究中具有前身ELANA冠状动脉吻合连接器6,7的OCT和SEM图像的例子证明了专利吻合而不缩小内膜增生的形成，并完全覆盖内皮，分别在6个月的随访（ 图9 </STRONG&gt;）。 <table border="0" cellpadding="0" cellspacing="0" width="432"><tbody><tr><td style="height:21px;width:345px;">吻合（N） </td><td style="width:87px;"> 3 </td></tr><tr><td> LITA（毫米，OD） </td><td> 3.2±0.2 </td></tr><tr><td> LAD（毫米，OD） </td><td> 1.8±0.0 </td></tr><tr><td>建设时间（min） </td><td> 3.4±0.4 * </td></tr><tr><td>襟翼检索率（％） </td><td> 100（3/3） </td></tr><tr><td>完全止血（％） </td><td> 100（3/3） </td></tr><tr><td>附加缝针</td><td> 0 </td></tr><tr><td>接枝基线流量（毫升/分钟） </td><td> 20±3 </td></tr><tr><td>在t = 5小时（毫升/分钟）接枝流</td><td> 18±5 </td></tr><tr><td>峰值流量充血反应（峰/基线流量） </td><td> 5.6±0.5 </td></tr></tbody></table> 表1:PI的手术数据大量的研究。数据以平均值±标准差或％（N）。 *包括:安装所述连接器，移植物冠状动脉，激光打孔动脉切开，远端接枝结扎连接。 <img alt="图1" src="/files/ftp_upload/52127/52127fig1highres.jpg" /> 图1:三位一体剪辑（A）动画图像冠状动脉吻合连接器，侧视图。该连接器被构造钛和适于靶冠状动脉与1.4和1.6毫米的内径（ID）。所述连接器包括:1:一个弹簧，它使2叉地打开和关闭，分别逐个（左下和右侧面板），通过将施加器到1的弹簧的轴（见星号，施加器不示出）。此外，它还提供了2叉的活性压缩2和<str翁&gt; 3:两个叉与每个2尖锐管脚，连接到所述弹簧（1）;上部叉（2，红色）将被插入到移植物，下部叉（3，蓝色）将被插入到冠状动脉4:一种血管外带（销2倍 ​​的厚度;在上透明地示出。面板），毗邻叉在他们的全长，在此获得额外的横向压缩。它被连接到弹簧，在锚点的叉之间。而打开下叉（3，蓝色;右下），上叉（2，红色）保持压缩（接枝）到带（4）。放大第（左上）表明在血管外带（4）。（B）的动画图象的连接器的，对角顶VI的前较长上部叉（2，红色）的前端的凹口的位置EW。上部叉子被打开（涂药未示出）。 <img alt="图2" src="/files/ftp_upload/52127/52127fig2highres.jpg" /> 图2:椭圆形的激光导管的椭圆形激光导管被用于激光打孔动脉切开术放入移植物和冠状动脉。请注意，激光导管不焊接或密封该吻合（A）中的外带（1;最宽的部分）。便于定位和稳定化入吻合连接器，并提供安全（ 即，它可防止通过连接器滑脱的激光导管并损坏冠状动脉的下壁）。所述真空通道（2）位于中央，并且由激光纤维（3）。（B）中 ，在激光导管的尖端的顶视图包围。 2激光纤维一排的可视化。 <img alt="图3" srC ="/文件/ ftp_upload / 52127 / 52127fig3highres.jpg"/&gt; 图3:该固定夹的外部，临时固定夹被用来固定和稳定激光导管插入装吻合连接器，确保了吻合施工期间导管的适当的垂直定位（A）的侧视图（左）和较差的视图（右）。所述弹簧（1）提供力到两个壳（2），其中保存了导管，并且该杆（3），它抓住连接器（B）中的箭头指向的导管，其垂直地通过固定夹迷恋并形成一个与所述连接器和所述接枝（未示出）以稳定复合物。 <img alt="图4" src="/files/ftp_upload/52127/52127fig4highres.jpg" /> 图4:喷头（A）。一个标准的动脉瘤夹器控制通过弹簧的下涂敷轴的上叉（见小节），此外，固定夹。（B）中的原型的Vasco敷贴控制通过弹簧的上应用的轴的下叉（请参阅本节）。 <img alt="图5" src="/files/ftp_upload/52127/52127fig5highres.jpg" /> 图5:用三一剪辑系统中的冠状动脉吻合过程冠状动脉吻合连接器（A）的安装:施放（未示出）用于插入所述连接器的上叉进入灌注移植物的管腔，定向远侧。注意:通过释放施加器，该连接器关闭，并且积极地压缩该2叉和血管外带之间的接枝（B）中的安装和激光打孔接枝。激光导管血管内导入，通过DISTA接枝，插入连接器的升自由端，并垂直地由所述外固定夹迷恋。接枝是激光打孔。在接枝（ 即 "瓣"）的激光打孔片段的箭头点（C）的接枝到冠状动脉的非阻塞性连接:施放（未示出）用于插入下部叉子。叉穿刺冠状动脉壁和被完全插入到灌注冠状动脉的内腔，定向远侧。在插入过程中，上叉保持移植到血管外频段压缩，确保这个动作在移植的正确固定，而固定夹可以确保激光导管的正确定位垂直（D）的冠状激光打孔动脉切开术动脉:所述连接器是关闭的，并在2管腔内叉和血管外频带之间压缩两容器壁（ 即，移植物和冠状动脉）。冠状动脉壁激光打孔By中的迷恋导管，垂直放置到冠状动脉墙。随后，固定夹被删除，导管缩回，包括所检索的翼板（见箭头）。（E）的最后吻合。的结扎血管夹放置在移植物的远端。注:完整的连接器保持原位吻合施工后不会被删除。 <img alt="图6" src="/files/ftp_upload/52127/52127fig6highres.jpg" /> 图6:三位一体剪辑术中通过时间的流量测量促进离开胸廓内动脉（LITA）-to-左 ​​前降支（LAD）吻合术和传统的手工缝合LITA至LAD吻合无论是促进（A）。与手工缝合（B）的 LITA至LAD旁路示出了正常的吸引力流动曲线，低于5一个PI和一个predomina<html> NT舒张移植充填（舒张期充盈[DF] 80％），以最小的收缩期峰值，暗示了专利冠状动脉移植。 <img alt="图7" src="/files/ftp_upload/52127/52127fig7highres.jpg" /> 图7:三位一体剪辑五周冠状动脉造影协助左胸廓内动脉（LITA）-to-左 ​​前降支（LAD）吻合（临床前研究的一个例子）（A）横向侧视图。结扎hemoclips被放置在LITA（1）的远端和近端天然LAD（2）。所述连接器（3）可以只看见在侧视图。注意，覆盖的叉和所述连接器由非不透射线物质的血管外频带能够被看见。所述LITA的前端没有填充相反，通过简化新内膜暗示重塑。（B）中的顶视图。 ghres.jpg"/&gt; 图8:治疗和三位一体剪辑重塑促进左胸廓内动脉（LITA）-to-左 ​​前降支（LAD）吻合:宏观和组织视图（A）从LAD内宏观上看，在5小时跟进。专利吻合，没有任何腔内血栓形成论证。连接器的下部叉子定位intraluminally，而不捕获或损坏的横向或下冠状动脉壁。两者LITA和LAD的小而尖的激光切割边缘（0.1毫米），是在叉之间可见的，并且这两个容器壁被定位恰在彼此的顶部上，不重叠外膜组织。（B）的宏观鉴于在LAD内以5周的随访（临床前研究的一个例子）。专利吻合证明和腔内叉和激光边缘被完全覆盖组织。层，而不缩小吻合口（C）的组织学横截面，中间吻合，在5周的随访（12.5X放大;临床前研究的一个例子）。由新内膜（NI）的连接器的简化覆盖（最初intraluminally曝光）叉（1）是可见的。放大第（40X放大倍率）演示缩回和改造激光边缘。叉和血管外带之间，动脉壁的压缩被看作无不良重构（ 例如，侵蚀，脱位，或形成假性）。此外，下壁不受影响，而没有任何内膜增生反应（可能是暗示对于[激光可]损坏），而且没有过度的炎性细胞反应被发现（这有可能由异物植入物被触发）。该LITA中，"死角"，的前端填充有有组织血栓，覆盖内膜组织，精简anastom肌病（未示出）。最后，将连接器的弹簧完全集成，extravascularly，所述LITA和LAD之间，无侵蚀作用或损坏相邻动脉壁（未示出）。注:下冠状动脉壁上的中断可能是检查前，纵向开放的冠状动脉。比例尺（1毫米）在左下角提供。 <img alt="图9" src="/files/ftp_upload/52127/52127fig9highres.jpg" /> 图9:右胸廓内动脉的冠状动脉内光学相干断层扫描（OCT）图像（RITA）的例子-to-右冠状动脉（RCA）吻合和扫描电子显微镜左胸廓内动脉（SEM）图像（LITA ）-to-左 ​​前降支（LAD）吻合。两人都构建了一个前身ELANA冠状动脉接头6,7。（A）OCT图像在6个月的随访，横截面，中anastomOSIS 7。注意:在OCT线是在管腔中可见。横向线表示的最小宽度吻合（= 2.2毫米）。 C =连接器; RITA =右胸廓内动脉; RCA =右冠状动脉。（B）的详细SEM图像，在连接器展示完全覆盖与内皮细胞（2,080X放大倍数）的叉的水平，在6个月随访6,7。

Watch this Scientific Journal Video about Evaluation of a Novel Laser-assisted Coronary Anastomotic Connector - the Trinity Clip - in a Porcine Off-pump Bypass Model at JoVE.com

Evaluation of a Novel Laser-assisted Coronary Anastomotic Connector - the Trinity Clip - in a Porcine Off-pump Bypass Model

This paper describes a novel nonocclusive coronary anastomotic connector in a porcine off-pump coronary artery bypass (OPCAB) model. This easy-to-use coronary connector has intrinsic potential to facilitate minimally invasive OPCAB surgery.

评价一种新型激光辅助冠状动脉吻合口连接器 - 三位一体夹 - 在猪非体外循环旁路模式

To simplify and facilitate beating heart (i.e., off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the ...

evaluation-novel-laser-assisted-coronary-anastomotic-connector

Research

JoVE Journal

Medicine

15.4K 次观看.  University Medical Center Utrecht. This paper describes a novel nonocclusive coronary anastomotic connector in a porcine off-pump coronary artery bypass (OPCAB) model. This easy-to-use coronary connector has intrinsic potential to facilitate minimally invasive OPCAB surgery.

视频: Evaluation of a Novel Laser-assisted Coronary Anastomotic Connector - the Trinity Clip - in a Porcine Off-pump Bypass Model

Sublingual Immunotherapy as an Alternative to Induce Protection Against Acute Respiratory Infections

Sublingual route has been widely used to deliver small molecules into the bloodstream and to modulate the immune response at different sites. It has been shown to effectively induce humoral and cellular responses at systemic and mucosal sites, namely the lungs and urogenital tract. Sublingual vaccination can promote protection against infections at the lower and upper respiratory tract; it can also promote tolerance to allergens and ameliorate asthma symptoms. Modulation of lung&#8217;s immune response by sublingual immunotherapy (SLIT) is safer than direct administration of formulations by intranasal route because it does not require delivery of potentially harmful molecules directly into the airways. In contrast to intranasal delivery, side effects involving brain toxicity or facial paralysis are not promoted by SLIT. The immune mechanisms underlying SLIT remain elusive and its use for the treatment of acute lung infections has not yet been explored. Thus, development of appropriate animal models of SLIT is needed to further explore its potential advantages. 
This work shows how to perform sublingual administration of therapeutic agents in mice to evaluate their ability to protect against acute pneumococcal pneumonia. Technical aspects of mouse handling during sublingual inoculation, precise identification of sublingual mucosa, draining lymph nodes and isolation of tissues, bronchoalveolar lavage and lungs are illustrated. Protocols for single cell suspension preparation for FACS analysis are described in detail. Other downstream applications for the analysis of the immune response are discussed. Technical aspects of the preparation of Streptococcus pneumoniae inoculum and intranasal challenge of mice are also explained.
SLIT is a simple technique that allows screening of candidate molecules to modulate lungs&#8217; immune response. Parameters affecting the success of SLIT are related to molecular size, susceptibility to degradation and stability of highly concentrated formulations.

The present work illustrates the convenience of using sublingual immunotherapy to boost the innate immune response in the lungs and confer protection against acute pneumococcal pneumonia in mouse.

舌下免疫治疗作为一种替代起到保护作用对急性呼吸道感染

Sublingual route has been widely used to deliver small molecules into the bloodstream and to modulate the immune response at different sites. It has been ...

科研

医学

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality of life when compared to dialysis. Although the last few decades have seen major improvements in patient outcomes following kidney transplantation, the increasing shortage of available organs represents a severe problem worldwide. To expand the donor pool, marginal kidney grafts recovered from extended criteria donors (ECD) or donated after circulatory death (DCD) are now accepted for transplantation. To further improve the postoperative outcome of these marginal grafts, research must focus on new therapeutic approaches such as alternative preservation techniques, immunomodulation, gene transfer, and stem cell administration.
Experimental studies in animal models are the final step before newly developed techniques can be translated into clinical practice. Porcine kidney transplantation is an excellent model of human transplantation and allows investigation of novel approaches. The major advantage of the porcine model is its anatomical and physiological similarity to the human body, which facilitates the rapid translation of new findings to clinical trials. This article offers a surgical step-by-step protocol for an autotransplantation model and highlights key factors to ensure experimental success. Adequate pre- and postoperative housing, attentive anesthesia, and consistent surgical techniques result in favorable postoperative outcomes. Resection of the contralateral native kidney provides the opportunity to assess post-transplant graft function. The placement of venous and urinary catheters and the use of metabolic cages allow further detailed evaluation. For long-term follow-up studies and investigation of alternative graft preservation techniques, autotransplantation models are superior to allotransplantation models, as they avoid the confounding bias posed by rejection and immunosuppressive medication.

Porcine models of organ transplantation provide an important platform to study mechanisms of organ preservation. This article describes a heterotopic porcine renal autotransplantation model, which allows investigating new approaches to improve the outcome of transplantation using marginal kidney grafts.

异位自体肾移植的猪模型:有步骤，分步协议

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality ...

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. ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

小鼠体外循环模型: 一种新方法

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

Surgical Swine Model of Chronic Cardiac Ischemia Treated by Off-Pump Coronary Artery Bypass Graft Surgery

Chronic cardiac ischemia that impairs cardiac function, but does not result in infarct, is termed hibernating myocardium (HM). A large clinical subset of coronary artery disease (CAD) patients have HM, which in addition to causing impaired function, puts them at higher risk for arrhythmia and future cardiac events. The standard treatment for this condition is revascularization, but this has been shown to be an imperfect therapy. The majority of pre-clinical cardiac research focuses on infarct models of cardiac ischemia, leaving this subset of chronic ischemia patients largely underserved. To address this gap in research, we have developed a well-characterized and highly reproducible model of hibernating myocardium in swine, as swine are ideal translational models for human heart disease. In addition to creating this unique disease model, we have optimized a clinically relevant treatment model of coronary artery bypass surgery in swine. This allows us to accurately study the effects of bypass surgery on heart disease, as well as investigate additional or alternate therapies. This model surgically induces single vessel stenosis by implanting a constrictor on the left anterior descending (LAD) artery in a young pig. As the pig grows, the constrictor creates a gradual stenosis, resulting in chronic ischemia with impaired regional function, but preserving tissue viability. Following the establishment of the hibernating myocardium phenotype, we perform off-pump coronary artery bypass graft surgery to revascularize the ischemic region, mimicking the gold-standard treatment for patients in the clinic.

This protocol presents a surgical large animal model of chronic, single vessel ischemia that results in regional abnormalities but does not create infarct, known as hibernating myocardium. Following establishment of chronic ischemia, animals are treated with off-pump LIMA-LAD coronary artery bypass graft surgery to revascularize the ischemic tissue.

非体外循环冠状动脉旁路移植术治疗慢性心肌缺血的外科猪模型

Chronic cardiac ischemia that impairs cardiac function, but does not result in infarct, is termed hibernating myocardium (HM). A large clinical subset of ...

Development of a Noninvasive, Laser-Assisted Experimental Model of Corneal Endothelial Cell Loss

Nd:YAG lasers have been used to perform noninvasive intraocular surgery, such as capsulotomy for several decades now. The incisive effect relies on the optical breakdown at the laser focus. Acoustic shock waves and cavitation bubbles are generated, causing tissue rupture. Bubble sizes and pressure amplitudes vary with pulse energy and position of the focal point. In this study, enucleated porcine eyes were positioned in front of a commercially available Nd:YAG laser. Variable pulse energies as well as different positions of the focal spots posterior to the cornea were tested. Resulting lesions were evaluated by two-photon microscopy and histology to determine the best parameters for an exclusive detachment of corneal endothelial cells (CEC) with minimum collateral damage. The advantages of this method are the precise ablation of CEC, reduced collateral damage, and above all, the non-contact treatment.

Here, we present a protocol to detach corneal endothelial cells (CEC) from Descemet&#8217;s membrane (DM) using a neodymium:YAG (Nd:YAG) laser as an ex vivo disease model for bullous keratopathy (BK).

角膜内皮细胞损失非侵入性激光辅助实验模型的发展

Nd:YAG lasers have been used to perform noninvasive intraocular surgery, such as capsulotomy for several decades now. The incisive effect relies on the ...

In Vitro Model of Coronary Angiogenesis

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects in these vessels represent severe health risks such as in atherosclerosis, which can lead to myocardial infarctions and heart failures in patients. Consequently, coronary artery disease is one of the leading causes of death worldwide. Despite its clinical importance, relatively little progress has been made on how to regenerate damaged coronary arteries. Nevertheless, recent progress has been made in understanding the cellular origin and differentiation pathways of coronary vessel development. The advent of tools and technologies that allow researchers to fluorescently label progenitor cells, follow their fate, and visualize progenies in vivo have been instrumental in understanding coronary vessel development. In vivo studies are valuable, but have limitations in terms of speed, accessibility, and flexibility in experimental design. Alternatively, accurate in vitro models of coronary angiogenesis can circumvent these limitations and allow researchers to interrogate important biological questions with speed and flexibility. The lack of appropriate in vitro model systems may have hindered the progress in understanding the cellular and molecular mechanisms of coronary vessel growth. Here, we describe an in vitro culture system to grow coronary vessels from the sinus venosus (SV) and endocardium (Endo), the two progenitor tissues from which many of the coronary vessels arise. We also confirmed that the cultures accurately recapitulate some of the known in vivo mechanisms. For instance, we show that the angiogenic sprouts in culture from SV downregulate COUP-TFII expression similar to what is observed in vivo. In addition, we show that VEGF-A, a well-known angiogenic factor in vivo, robustly stimulates angiogenesis from both the SV and Endo cultures. Collectively, we have devised an accurate in vitro culture model to study coronary angiogenesis.

In vitro models of coronary angiogenesis can be utilized for the discovery of the cellular and molecular mechanisms of coronary angiogenesis. In vitro explant cultures of sinus venosus and endocardium tissues show robust growth in response to VEGF-A and display a similar pattern of COUP-TFII expression as in vivo.

冠状血管生成体模型

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects ...

A Modified Two Kidney One Clip Mouse Model of Renin Regulation in Renal Artery Stenosis

Renal artery stenosis is a common condition in patients with coronary or peripheral vascular disease where the renin angiotensin aldosterone system (RAAS) is overactivated. In this context, there is a narrowing of the renal arteries that stimulate an increase in the expression and release of renin, the rate-limiting protease in RAAS. The resulting rise in renin expression is a known driver of renovascular hypertension, frequently associated with kidney injury and end organ damage. Thus, there is a great interest in developing novel treatments for this condition. The molecular and cellular mechanism of renin control in renal artery stenosis is not fully understood and warrants further investigation. To induce renal artery stenosis in mice, a modified 2 kidney 1 clip (2K1C) Goldblatt mouse model was developed. The right kidney was stenosed in wild type mice and sham operated mice were used as control. After renal artery stenosis, we determined renin expression and kidney injury. Kidneys were harvested, and fresh cortices were used to determine protein and mRNA expression of renin. This animal model is reproducible and can be used to study pathophysiological responses, molecular and cellular pathways involved in renovascular hypertension and kidney injury.

A modified 2 kidney 1 clip (2K1C) Goldblatt mouse model was developed using polyurethane tubing to initiate renal artery stenosis, inducing an increase in renin expression and kidney injury. Here, we describe a detailed procedure of preparing and placing the cuff onto the renal artery to generate a reproducible and consistent 2K1C mouse model.

肾动脉狭窄肾素调节的改良双肾单夹小鼠模型

Renal artery stenosis is a common condition in patients with coronary or peripheral vascular disease where the renin angiotensin aldosterone system (RAAS) ...

Retinal Pathophysiological Evaluation in a Rat Model

A posterior segment eye disease like diabetic retinopathy alters the physiology of the retina. Diabetic retinopathy is characterized by a retinal detachment, breakdown of the blood-retinal barrier (BRB), and retinal angiogenesis. An in vivo rat model is a valuable experimental tool to examine the changes in the structure and function of the retina. We propose three different experimental techniques in the rat model to identify morphological changes of retinal cells, retinal vasculature, and compromised BRB. Retinal histology is used to study the morphology of various retinal cells. Also, quantitative measurement is performed by retinal cell count and thickness measurement of different retinal layers. A BRB breakdown assay is used to determine the leakage of extraocular proteins from the plasma to vitreous tissue due to the breakdown of BRB. Fluorescence angiography is used to study angiogenesis and leakage of blood vessels by visualizing retinal vasculature using FITC-dextran dye.

Diabetic retinopathy is one of the leading causes of blindness. Histology, blood-retinal barrier breakdown assay, and fluorescence angiography are valuable techniques to understand the pathophysiology of the retina, which could further enhance the efficient drug screening against diabetic retinopathy.

大鼠模型中的视网膜病理生理学评估

A posterior segment eye disease like diabetic retinopathy alters the physiology of the retina. Diabetic retinopathy is characterized by a retinal ...

Establishment and Evaluation of a Porcine Vein Graft Disease Model

Venous graft disease (VGD) is the leading cause of coronary artery bypass graft (CABG) failure. Large animal models of CABG-VGD are needed for the investigation of disease mechanisms and the development of therapeutic strategies. 
To perform the surgery, we enter the cardiac chamber through the third intercostal space and carefully dissect the internal mammary vein and immerse it in normal saline. The right main coronary artery is then treated for ischemia. The target vessel is incised, a shunt plug is placed, and the distal end of the graft vein is anastomosed. The ascending aorta is partially blocked, and the proximal end of the graft vein is anastomosed after perforation. The graft vein is checked for patency, and the proximal right coronary artery is ligated. 
CABG surgery is performed in minipigs to harvest the left internal mammary vein for its use as a vascular graft. Serum biochemical tests are used to evaluate the physiological status of the animals after surgery. Ultrasound examination shows that the proximal, middle, and distal end of the graft vessel are unobstructed. In the surgical model, turbulent blood flow in the graft is observed upon histological examination after the CABG surgery, and venous graft stenosis associated with intimal hyperplasia is observed in the graft. The study here provides detailed surgical procedures for the establishment of a repeatable CABG-induced VGD model.

In this protocol, novel pig vein bypass grafting was performed through a small incision in the left chest wall without cardiopulmonary bypass. A postoperative pathology study was done, which showed intimal thickening.