这项研究是由Lundbeck公司基金会，克伦Elise的詹森基金会，丹麦中部地区的卫生研究基金和奥胡斯大学医院临床医学研究所的财政支持。我们承认唐纳德F史密斯，安德斯斯蒂恩Soerensen，亨里克Soerensen，Asbjoern Ettrup - 克里斯滕森，并在Paaskehoejgaard工作人员娴熟的援助表示感谢。

1。插管<ol><li>气管插管所需的设备：辅助通气，打开口，先端钝低压吸，直叶片兽医喉镜在17和25厘米长之间，钢丝，与空气的注射器，听诊器和胶带吊索的手段。成人哥廷根小型猪气管插管大小5至7毫米的10。在这个视频使用一个5.5毫米的管10 。 </li><li>小型猪是放置在它的后面，并得到适当的支持，以确保喉通道保持伸直。 </li><li>稍微拉出舌头和下颚由一名助理举行开放而不过度的头部。航空公司或过度扩展头的旋转，可以堵塞呼吸道，喉开放更加难以确定和复杂管气管位置。 </li><li>喉镜是通过咽腔和使用，以压低舌头，使会厌可见。吸的一角是用来取代软腭会厌。喉镜的一角，然后按舌根会厌向上走向，揭示了喉开放。 </li><li>映入眼帘的arytenoids软骨和声带进入气管，气管插管是先进的，有轻微的旋转轻轻期间届满。 </li><li>套管是按照制造规格膨胀，同时照顾，以避免过量的通货膨胀，这可能引起肿胀和水肿的航空公司。猪有支气管右颅pulmonal叶通风。因此重要的是气管分叉以上地方管。这是促进使用正确的管大小，重要的是使用胶带口鼻水平紧固管近端。 </li><li>小型猪是正确插管时，感受到的空气的自由通行，必须通过管，并应显示在胸廓的左侧和右侧胸部听诊呼吸音。 </li><li>连接与监控系统的小型猪呼吸后，生理呼气二氧化碳值应该是可以衡量和二氧化碳曲线如下的呼吸模式，可以观察到呼吸 6,10设置决定。 </li><li>二氧化碳和血氧饱和度值最初可能由术前用药和麻醉诱导的影响。然而，这些值应尽快恢复正常，适当的通风 6,10 。尽管人工辅助通气，紫绀的任何迹象表明管放置不当。最常见的错误是无心管放置在食道。 </li></ol> 2。经尿道膀胱导尿<ol><li>此过程只适用于女性的猪。对于成年小型猪的一个尿管法国大小8个或10个是适当的。由于曲率和尿道外口的大小，与钢丝导管是可取的。可用于小型小型猪鼻窥器窥器而具有更长的叶片可用于大型动物。必要的设备还包括一个钝头镊子，注射器，生理盐水，润滑剂和一个封闭的系统尿液引流袋。 </li><li>为了更好地可视化在导尿尿道外口的小型猪是放置在它的后面的下腰地区的支持，而一个助理拉后腿颅。 </li><li>润滑窥器插入，打开可视化的阴道壁。女性尿道外口位于阴道内的子宫颈的距离（ 图1A）的一半大约三分之一的地板。不过，因为动物是放在后面，尿道口的位置会出现倒置。 </li><li>钝尖镊子控制导管，使导管尖端在继续钳。在尿道口（ 图1B）小心地插入导管。如果用力过猛，在使用导尿，尿道很容易受到创伤。然而，最初可能会感觉到轻微的抵抗，直到导管已通过外部尿道括约肌。如果这个问题仍然存在，这可能表明，动物是不足够深镇静剂。解决的办法是等待一段时间，让麻醉的效果。 </li><li>由于导管到达膀胱，取出钢丝，尿便看到流淌。 </li><li>用生理盐水的气球充气按照制造规格，直到气球渔获膀胱颈部，轻轻一拉导管。 </li><li>将封闭系统尿液引流袋。 </li><li>带尾巴导管的导管，以避免意外的位移时，处理的动物。 </li></ol> 3。股动脉和静脉catheterizatioñ <ol><li>必要的设备包括：手术刀，手术剪钝尖，组织钳，小钝尖手术钳，小自护组织拉钩，持针器，手术掉期，用针缝合，肝素生理盐水作为冲洗液，以保持尿管通畅两个静脉插管，18号两个4法国与导引，并采用Seldinger导丝BRITE提示鞘。 </li><li>测试启动程序之前，趾间疼痛反射，应该评估麻醉深度。 </li><li>股动脉和静脉是在它的后面放置的小型猪接触和后排的腿部横向翻供。确定在皮肤皱褶之间的股薄肌和缝匠肌隐动脉内侧表面的脉动消失。 </li><li>作一纵形的浅表性皮肤切口，颅这一点，从而避免无意中破坏内侧隐血管。 </li><li>使用钝头剪刀，解剖的基础皮下组织。 </li><li>内侧隐血管，先用小钝尖手术钳，然后数字穿透部位分为颅缝匠肌及股薄肌的筋膜分工。 </li><li>这两个肌肉群是一个小的自护组织拉钩分开，同时注意不要损伤股神经和血管。 </li><li>使用钝性分离约1至2厘米长的动脉被隔离。 </li><li>旋转的斜面venflon针，使管腔朝上。弯曲的针一点点，所以它会按照船只曲率更容易。请记住，以确保针仍然可以毫不费力地来回移动。 </li><li>动脉被刺破，针缩回，采用Seldinger导丝通过venflon管插入。 </li><li>删除venflon管，一边轻轻地运用对动脉的压力，以确保导丝固定。 </li><li>与导引鞘插入在采用Seldinger线和先进到所需的位置，被删除后，介绍人，并采用Seldinger线。 </li><li>股静脉位于正下方，内侧动脉。隔离静脉使用钝性分离后，静脉插管，动脉。 </li><li>为了验证，无论是动脉和静脉插管是否正确，从中既鞘的​​血液和比较样品的颜色。每当绘制的血液样本，记得后来与肝素生理盐水冲洗导管，以保持尿管通畅。 </li><li>导管担保和关闭几个缝合皮肤。 </li><li>一艘环可放置周围的动脉和静脉，以注视穿刺前的船只。尽管它的优势，我们一般避免这种额外的步骤，因为它需要过剩剥离的船只，可以增加损害横向和深分行的风险。 </li></ol> 4。 Transcardial灌注<ol><li>需要以下设备的程序：灌注系统（ 图2），有关灌注液，手术刀，手术剪钝尖，骨剪，和两个大的自我持物钳，其中至少有一个有一个弯曲的技巧。自护胸骨牵开也不是绝对必要的，但它简化了程序，大大。 </li><li>必须在通风良好的室内，一个大功率的排烟系统和设施，使随后的安全处置收集和血液灌流液体Transcardial灌注。 </li><li>静脉点滴注射致死剂量的苯巴比妥后，测试趾间疼痛反射，然后再继续。 </li><li>心脏和主动脉的最佳途径是通过胸骨正中。作深刻的纵向切开皮肤，从胸骨柄剑突延伸。增益通过一个小切口在剑突膜片略低于，进入胸腔。 </li><li>插入胸腔骨剪刀和对开胸骨。执行此过程时，必须采取护理，在胸骨内的一些地方，因为可能是心脏壁。因此，作出一定的骨剪，刀片在所有时间内对胸腔胸骨内掌握。 </li><li>胸骨后进行了划分，把自护的胸骨牵开，打开胸腔。 </li><li>如果仍然完好，在心包小切口在心尖和数字，然后打开心包囊。 </li><li>识别左心室，右耳廓和主动脉，加上优越腔静脉（ 图 1C ） </li><li>心尖​​附近的一个小表面的剪切和访问左心室心肌射孔与钝尖仪器。 </li><li>插入的perfus离子左心室内插管，主动脉插管尖端移动颅。 </li><li>当导管是在主动脉的手指之间的感觉，到位与自我控股弯钳钳套管。 </li><li>做一个切口在右耳廓，并开始通过灌注导管注入固定液。主动脉灌流在流可以感受到，血液倒入通过切开右耳廓进入小型猪血管灌流流继续。请注意如何系统性与多聚甲醛灌注诱导过度伸展四肢和由于醛交联的神经和肌肉的浅表肌肉抽搐。当使用压缩空气代替重力灌注系统供电，确保取出导管，或钳明确的硅胶管，前罐是空的，以避免空气进入血管系统。 </li></ol><img src="/files/ftp_upload/2652/2652fig1.jpg" alt="图1" /> 图1。答：哥廷根小型猪的女性尿道外开放B：导管插入后尿道口：后正中切口小型猪心脏的腹面观。 <img src="/files/ftp_upload/2652/2652fig2.jpg" alt="图2" /> 图2。答：灌注系统由入口管可调压力阀和一个杠杆，用来控制压缩空气入口控股灌流液的容器。清除硅胶管从4毫米的钢管的钝尖的导管连接容器，B和C。通过压缩进口杆，压力容器和灌流上升是通过导管ð推进：当明确硅胶管和套管填充与灌流，流量是停止与钳系统已准备就绪。

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丹麦实验动物督察，丹麦司法部批准的所有程序，并根据（美国国立卫生研究院出版86-23号，1985年修订）“实验动物保健的原则”进行。

此演示文稿提供了有关如何执行四个经常使用的程序，在小型猪研究12,13的视觉指示一步一步。描述的技术可以用很少或根本没有适应，长白猪的猪和其他大型研究的动物如狗和羊14。必须采用无菌技术操作，以避免感染，当生存研究中所描述的程序。 麻醉诱导是很重要的动物福利和科学的原因，并应提供角膜反射和良好的肌肉松弛的情况下，气管插管前尝试。然而，因为使用的麻醉剂诱导是不给予固定剂量，但预期的效果观察，直到管理，重要的是密切监测动物因窒息的危险，特别是如果使用异丙酚6,10。 xylocaine喷雾或肌肉松弛剂的使用气管插管，可以方便的引入管，但神经肌肉阻断剂的使用应只能由那些训练有素的这种程序，以符合道德要求的人员进行。应注意不要创伤插管时喉通道，以避免气管肿胀和水肿。 10甾体抗炎药或类似的消炎药物可以治疗呼吸道水肿。猪气管插管，可以从不同的位置，但动物在它的后面往往是定期开展人力插管容易，因为它提出以类似的方式6呼吸道。插管过程中最常见的错误是无心管放置在食管，喉通道时，动物是在它的后面正好低于位于。请注意，不应该管的先进没有首先清晰地识别arytenoids软骨和声带。 经尿道经尿道膀胱导尿是在女猪很容易做到，而这个过程是非常困难的，也许是不可能的，在男性6，的。应该指出，因为女性的尿道口阴道前，阴道口较大，可误插管。灭菌和消毒润滑剂的使用是最好的生存研究。此外，局部麻醉作用的润滑剂，可缓解导尿过程。 当执行股动脉和静脉catherization，血管导管的选择和如何担保非生存和生存研究之间的差异，尤其是当导管植入时间6长时间。例如慢性植入导管应高度生物相容性，有固定的保留或袖口珠，以及不随着 ​​时间的推移创伤的血管壁，从而诱发血栓形成或侵蚀6。 在目前的视频中，我们使用一个封闭的系统，由压缩空气驱动transcardial灌注。另一种选择是使用由重力驱动系统。重力系统，压缩空气系统利用比较优势的是，他们是简单，高效，目前没有加压空气进入血管系统的风险。然而，重力系统施加静水压力不断变化，在灌注过程中，这可能会影响的 15个组织学结果。根据一个要执行的组织病理学研究哪种类型，它可能是有利的肝素生理盐水灌注与动物的。这是特别importantance进行免疫荧光染色时，为了避免红细胞引起的自体荧光的问题。此外，如果感兴趣的器官是大脑，可以钳制降主动脉保存固定液。然而，在猪，这一步需要额外的解剖，并将延长的过程。

<table border="1"><tbody><tr><th>程序</th><th>设备</th><th>公司</th><th>目录号</th></tr><tr><td>插管</td><td></td><td></td><td></td></tr><tr><td></td><td>呼吸器</td><td>德恩的S / AVANCE，GE医疗集团</td><td> 1009-9002-000 </td></tr><tr><td></td><td>先端钝的吸</td><td> Unomedical </td><td> 34091181 </td></tr><tr><td></td><td>喉镜</td><td>磨坊主</td><td> 85-0045（喉镜集） </td></tr><tr><td></td><td>气管导管5.5 mm内径</td><td>史密斯医疗国际有限公司</td><td> 100/199/055 </td></tr><tr><td></td><td>短剑</td><td>哈德逊RCI </td><td> 5-151001 </td></tr><tr><td></td><td>注射器5毫升</td><td></td><td></td></tr><tr><td></td><td>听筒</td><td>马丁</td><td> 17-262-00 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>膀胱catherization </td><td></td><td></td><td></td></tr><tr><td></td><td> AquaFlate甘油硅胶导尿管8，法国与钢丝</td><td>鲁施</td><td> 178003 </td></tr><tr><td></td><td>窥鼻</td><td>马丁</td><td> 36-859-50 </td></tr><tr><td></td><td>窥器直肠</td><td>马丁</td><td> 30-613-15 </td></tr><tr><td></td><td>钝尖镊子</td><td>马丁</td><td> 13-374-16 </td></tr><tr><td></td><td>导管润滑凝胶</td><td></td><td></td></tr><tr><td></td><td> Cathejell利多卡因</td><td> Montavit </td><td> CJL08501 </td></tr><tr><td></td><td> Instillagel梳子。 </td><td> FARCO制药</td><td> 08-40-200 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td></td><td> A4尿液引流袋</td><td> Unomedical </td><td> 38 155 1​​S </td></tr><tr><td></td><td>胶带</td><td></td><td></td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>股动脉和静脉catherization </</td><td></td><td></td><td></td></tr><tr><td></td><td>解剖刀</td><td>马丁</td><td> 10-295-11 </td></tr><tr><td></td><td>钝尖手术剪</td><td>马丁</td><td> 11-881-18 </td></tr><tr><td></td><td>组织钳</td><td>马丁</td><td> 12-100-14 </td></tr><tr><td></td><td>小钝尖手术钳</td><td>马丁</td><td> 13-375-13 </td></tr><tr><td></td><td>小自护组织拉钩</td><td>马丁</td><td> 15-753-13 </td></tr><tr><td></td><td> Needleholder </td><td>马丁</td><td> 20-634-14 </td></tr><tr><td></td><td>手术掉期</td><td></td><td></td></tr><tr><td></td><td>缝合用针</td><td></td><td></td></tr><tr><td></td><td>盐水</td><td></td><td></td></tr><tr><td></td><td> Heparine </td><td>低轨</td><td> 585679 </td></tr><tr><td></td><td> 18压力表venflon </td><td> BD Venflon临</td><td> 393206 </td></tr><tr><td></td><td> 4法国光亮与导引，并采用Seldinger导丝的尖端护套</td><td>科迪斯</td><td> 154-2629-1/0017 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Transcardial灌注</td><td></td><td></td><td></td></tr><tr><td></td><td>加压灌注系统</td><td>从地区最专业的五金商店取得的首页</td><td></td></tr><tr><td></td><td>排烟系统</td><td>革兰氏A / S公司</td><td> 01750000和0303700 </td></tr><tr><td></td><td> Perfusant 5L </td><td></td><td></td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td></td><td>钝尖手术剪</td><td>马丁</td><td> 11-881-18 </td></tr><tr><td></td><td>骨剪</td><td>福斯特</td><td> 16150-24 </td></tr><tr><td></td><td>大弯尖自我持物钳</td><td>马丁</td><td> 13-421-18 </td></tr><tr><td></td><td>自护的胸骨牵开</td><td>鲁道夫医疗</td><td> 7520-10 </td></tr><tr><td></td><td>戊巴比妥</td><td> TheRoyal兽医和农业大学（KVL） </td><td> 200毫克/毫升。 </td></tr><tr><td></td><td> 22压力表venflon </td><td> BD Venflon临</td><td> 393202 </td></tr></tbody></table>

basic surgical techniques in the g&ouml;ttingen minipig: intubation, bladder catheterization, femoral vessel catheterization, and transcardial perfusion

哥廷根小型猪研究的主题，如神经科学，毒理学，糖尿病，肥胖，和实验手术的出现，反映了人体解剖学和生理学1-6这些动物非常相似。哥廷根小型猪的大小允许使用的手术6-8在人类所使用的类似设备和先进的成像方式。本教学视频的目的是演示如何进行气管插管，经尿道膀胱插管，股动脉和静脉插管，以及transcardial灌注，增加对小型猪在生物医学研究的价值的意识。 每当一个小型猪进行全身麻醉，因为它维护了专利的气道，允许辅助通气，保护吸呼吸道，应进行气管插管。经尿道膀胱导尿，可在漫长的手术过程中提供有关水合状态的有用信息，以及肾脏和心血管功能。此外，尿导管检查可以防止污染微妙的医疗技术设备和膀胱疼痛的扩展，这可能会伤害动物和不必要的影响的实验，由于迷走神经张力增加和生理参数的改变。动脉和静脉插管获得重复的血液样本，并监测各种生理参数。涉及基于帧的立体定向神经外科和脑成像进行实验时，股血管插管导尿颈部血管四通八达的交通网络，更可取。执行船只在生存研究导尿时，必须严格无菌技术操作，以避免感染6。 Transcardial灌注是最有效的固定方法，并准备小型猪器官组织学和组织化学 2,9时，会产生杰出的成果。有关麻醉，手术和实验技术，在一般的猪的详细信息，我们是指骗取2007年。补充资料，对术前用药和麻醉诱导，辅助通气，镇痛，术前和术后护理哥廷根小型猪可通过<a href="http://www.minipigs.com">互联网</a> http://www.minipigs.com 10 。有关猪解剖的广泛的信息，我们是指镍等。卷1-5 11。

Watch this Scientific Journal Video about Basic Surgical Techniques in the GÃ¶ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion at JoVE

Basic Surgical Techniques in the G&amp;#246;ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

使用科学的国内和小型猪在近年来有明显的增加。演示如何执行，经尿道膀胱导尿插管，股动脉和静脉插管，以及transcardial灌注，我们的目标，进一步加大在生物医学研究中的哥廷根小型猪的价值。

基本外科技术，在哥廷根微型猪：气管插管，膀胱导尿，股血管的导管和Transcardial灌注

The emergence of the G&ouml;ttingen minipig in research of topics such as neuroscience, toxicology, diabetes, obesity, and experimental surgery reflects ...

basic-surgical-techniques-gttingen-minipig-intubation-bladder

Research

JoVE Journal

Medicine

Basic Surgical Techniques in the G&ouml;ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

69.0K 次观看.  Aarhus University Hospital. 使用科学的国内和小型猪在近年来有明显的增加。演示如何执行，经尿道膀胱导尿插管，股动脉和静脉插管，以及transcardial灌注，我们的目标，进一步加大在生物医学研究中的哥廷根小型猪的价值。

视频: Basic Surgical Techniques in the G&#246;ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

股动脉和静脉导管采血，药品监督管理局和意识的血压和心率测量

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

科研

医学

Cardiac Stress Test Induced by Dobutamine and Monitored by Cardiac Catheterization in Mice

Dobutamine is a &beta;-adrenergic agonist with an affinity higher for receptor expressed in the heart (&beta;1) than for receptors expressed in the arteries (&beta;2). When systemically administered, it increases cardiac demand. Thus, dobutamine unmasks abnormal rhythm or ischemic areas potentially at risk of infarction. 
Monitoring of heart function during a cardiac stress test can be performed by either ecocardiography or cardiac catheterization. The latter is an invasive but more accurate and informative technique that the former.
Cardiac stress test induced by dobutamine and monitored by cardiac catheterization accomplished as described here allows, in a single experiment, the measurement of the following hemodynamic parameters: heart rate (HR), systolic pressure, diastolic pressure, end-diastolic pressure, maximal positive pressure development (dP/dtmax) and maximal negative pressure development (dP/dtmin), at baseline conditions and under increasing doses of dobutamine.
As expected, in normal mice we observed a dobutamine dose-related increase in HR, dP/dtmax and dP/dtmin. Moreover, at the highest dose tested (12 ng/g/min) the cardiac decompensation of high fat diet-induced obese mice was unmasked.

We describe the protocol to perform a cardiac stress test induced by dobutamine and monitored by cardiac catheterization in normal mice. Also we show its application to unmask subclinical cardiac disease in high fat diet-induced obese mice.

多巴酚丁胺诱导的心脏压力测试和监控通过在小鼠心脏导管插入术

Dobutamine is a &beta;-adrenergic agonist with an affinity higher for receptor expressed in the heart (&beta;1) than for receptors expressed in the ...

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. Intra-tracheal inoculation of a known amount of bacteria has been successfully used to reproduce the pathogenesis of pneumonia which then develops into sepsis. Monitoring hemodynamic parameters and providing standard clinical treatment including infusion of antibiotics, fluids and drugs to maintain blood pressure is critical to simulate routine supportive care in this model but to do so requires both arterial and venous vascular access. The video details the surgical technique for implanting carotid artery and common jugular vein catheters in an anesthetized rat. Following a 72 hr recovery period, the animals will be re-anesthetized and connected to a tether and swivel setup attached to the rodent housing which connects the implanted catheters to the hemodynamic monitoring system. This setup allows free movement of the rat during the study while continuously monitoring pressures, infusing fluids and drugs (antibiotics, vasopressors) and performing blood sampling.

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

颈动脉和颈静脉导管在有意识大鼠模型进行血流动力学措施，输液和血液采样

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

Cardiac Catheterization in Mice to Measure the Pressure Volume Relationship: Investigating the Bowditch Effect

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these strategies is to examine their effects on heart function. There are several techniques to measure in vivo cardiac mechanics (e.g., echocardiography, pressure/volume relations, etc.). Compared to echocardiography, real-time left ventricular (LV) pressure/volume analysis via catheterization is more precise and insightful in assessing LV function. Additionally, LV pressure/volume analysis provides the ability to instantaneously record changes during manipulations of contractility (e.g., &#946;-adrenergic stimulation) and pathological insults (e.g., ischemia/reperfusion injury). In addition to the maximum (+dP/dt) and minimum (-dP/dt) rate of pressure change in the LV, an accurate assessment of LV function via several load-independent indexes (e.g., end systolic pressure volume relationship and preload recruitable stroke work) can be attained. Heart rate has a significant effect on LV contractility such that an increase in the heart rate is the primary mechanism to increase cardiac output (i.e., Bowditch effect). Thus, when comparing hemodynamics between experimental groups, it is necessary to have similar heart rates. Furthermore, a hallmark of many cardiomyopathy models is a decrease in contractile reserve (i.e., decreased Bowditch effect). Consequently, vital information can be obtained by determining the effects of increasing heart rate on contractility. Our and others data has demonstrated that the neuronal nitric oxide synthase (NOS1) knockout mouse has decreased contractility. Here we describe the procedure of measuring LV pressure/volume with increasing heart rates using the NOS1 knockout mouse model.

This article describes the measurement of murine left ventricular function via pressure/volume analysis at different heart rates.

心导​​管在小鼠测量压力体积的关系：调查的影响鲍迪奇

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these ...

Ex Situ Normothermic Machine Perfusion of Donor Livers

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous perfusion of organs provides the opportunity to improve organ quality and allows ex situ viability assessment of donor livers prior to transplantation. This video article provides a step by step protocol for ex situ normothermic machine perfusion (37 &#176;C) of human donor livers using a device that provides a pressure and temperature controlled pulsatile perfusion of the hepatic artery and continuous perfusion of the portal vein. The perfusion fluid is oxygenated by two hollow fiber membrane oxygenators and the temperature can be regulated between 10 &#176;C and 37 &#176;C. During perfusion, the metabolic activity of the liver as well as the degree of injury can be assessed by biochemical analysis of samples taken from the perfusion fluid. Machine perfusion is a very promising tool to increase the number of livers that are suitable for transplantation.

Ex Situ Normothermic Machine Perfusion of Donor Livers

Here we present a protocol describing oxygenated ex situ machine perfusion of donor liver grafts. This article contains a step by step protocol to procure and prepare the liver graft for machine perfusion, prepare the perfusion fluid, prime the perfusion machine and perform oxygenated normothermic machine perfusion of the liver graft.

供肝易地常温灌注机

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous ...

Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs

Catheterization of the intestinal lymph trunk in neonatal pigs is a technique allowing for the long-term collection of large quantities of intestinal (central) efferent lymph. Importantly, the collection of central lymph from the intestine enables researchers to study both the mechanisms and lipid constitutes associated with lipid metabolism, intestinal inflammation and cancer metastasis, as well as cells involved in immune function and immunosurveillance. A ventral mid-line surgical approach permits excellent surgical exposure to the cranial abdomen and relatively easy access to the intestinal lymph trunk vessel that lies near the pancreas and the right ventral segment of the portal vein underneath the visceral aspect of the right liver lobe. The vessel is meticulously dissected and released from the surrounding fascia and then dilated with sutures allowing for insertion and subsequent securing of the catheter into the vessel. The catheter is exteriorized and approximately 1 L/24 hr of lymph is collected over a 7 day period. While this technique enables the collection of large quantities of central lymph over an extended period of time, the success depends on careful surgical dissection, tissue handling and close attention to proper surgical technique. This is particularly important with surgeries in young animals as the lymph vessels can easily tear, potentially leading to surgical and experimental failure. The video demonstrates an excellent surgical technique for the collection of intestinal lymph.

We present a surgical procedure to catheterize the intestinal lymph trunk in neonatal pigs to collect large quantities of lipid metabolism components from efferent lymph.

在新生儿猪淋巴的肠淋巴躯干和收集的长期导尿

Catheterization of the intestinal lymph trunk in neonatal pigs is a technique allowing for the long-term collection of large quantities of intestinal ...

Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension

In this protocol, PAH is induced by combining a 60 mg/kg monocrotalin (MCT) injection with increased pulmonary blood flow through an aorto-caval shunt (MCT+Flow). The shunt is created by inserting an 18-G needle from the abdominal aorta into the adjacent caval vein. Increased pulmonary flow has been demonstrated as an essential trigger for a severe form of PAH with distinct phases of disease progression, characterized by early medial hypertrophy followed by neointimal lesions and the progressive occlusion of the small pulmonary vessels. To measure the right heart and pulmonary hemodynamics in this model, right heart catheterization is performed by inserting a rigid cannula containing a flexible ball-tip catheter via the right jugular vein into the right ventricle. The catheter is then advanced into the main and the more distal pulmonary arteries. The histopathology of the pulmonary vasculature is assessed qualitatively, by scoring the pre- and intra-acinar vessels on the degree of muscularization and the presence of a neointima, and quantitatively, by measuring the wall thickness, the wall-lumen ratios, and the occlusion score.

This protocol describes a surgical procedure to create a model for flow-induced pulmonary arterial hypertension (PAH) in rats and the procedures to analyze the principle hemodynamic and histological end-points in this model.

分流手术，右心导管，血管形态计量学在大鼠模型中的流量引起的肺动脉高压

In this protocol, PAH is induced by combining a 60 mg/kg monocrotalin (MCT) injection with increased pulmonary blood flow through an aorto-caval shunt ...

Harvest of Endothelial Cells from the Balloon Tips of Swan-Ganz Catheters after Right Heart Catheterization

A variety of pathologies lead to pulmonary hypertension (PH), which is defined as a mean pulmonary artery pressure exceeding 25 mmHg at rest. To further diagnose and manage PH, patients undergo repeated right heart catheterizations (RHC) wherein a Swan-Ganz catheter is advanced into a branch of the pulmonary artery and a balloon is inflated to wedge the catheter tip. This article illustrates a protocol whereby pulmonary artery endothelial cells (PAECs) may be harvested from the balloon tips of Swan-Ganz catheters after RHC, and purified with an anti- CD146 affinity column technique to purify putative PAECs. These cells might be used to provide an in situ snapshot of the biological state of the pulmonary vasculature endothelium to complement hemodynamic measurements obtained during RHC. Harvested and purified PAECs may be used for either cell culture or for subsequent analytical assays such as flow cytometery.

This protocol describes the removal and purification of pulmonary artery endothelial cells (PAECs) that adhere to the balloon tips of Swan-Ganz catheters while they are wedged in the pulmonary artery during right heart catheterization and remain adherent to the deflated balloon following removal from the patient's body.

右心导管插入术后天鹅管囊小贴中内皮细胞的提取

A variety of pathologies lead to pulmonary hypertension (PH), which is defined as a mean pulmonary artery pressure exceeding 25 mmHg at rest. To further ...

Brachial Artery Catheterization in Swine

The video describes in detail the catheterization of the distal brachial artery in swine. This technique enables researchers to measure arterial blood pressure continuously and collect arterial blood samples to assess arterial blood gas measurements. Arterial blood pressures and arterial blood gases are important physiological parameters to monitor during experimental procedures. In swine, four common methods of arterial catheterization have been described, including catheterization of the carotid, femoral, auricular, and medial saphenous arteries. Each of these techniques have advantages, such as ease of access for the auricular artery, and disadvantages that include deep tissue dissection for carotid artery catheterization. The described alternative method of arterial catheterization in swine, the catheterization of the distal aspect of the brachial artery, is a rapid procedure that requires relatively minimal tissue dissection and provides information that is in line with data collected from other arterial catheterization sites. The procedure uses a medial approach along an oblique plane of the lower brachium, positioned between the olecranon and the flexor aspect of the elbow joint, and this approach allows researchers the major advantage of unimpeded freedom for procedures that involve the caudoventral, caudodorsal back, or hind limbs of the pig. Due to the location of the upper forelimb of the catheterized vessel and potential challenges of effective homeostasis following catheter removal from the artery, this technique may be limited to non-recovery procedures.

The video describes in detail the catheterization of the distal brachial artery in swine. This procedure accurately measures arterial blood pressure and is a simple and fast method to collect samples for arterial blood gas measurements.

猪的臂动脉插管

The video describes in detail the catheterization of the distal brachial artery in swine. This technique enables researchers to measure arterial blood ...

Repeated Orotracheal Intubation in Mice

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ventral neck for direct confirmation of cannula placement in the trachea. The relative difficulty or the tissue trauma induced to the subject by such procedures can be an impediment to an investigator&#8217;s ability to perform longitudinal studies. This article illustrates a technique in which physical manipulation of the mouse following the use of a depilatory to remove hair from the ventral neck permits transcutaneous visualization of the trachea for orotracheal intubation regardless of degree of skin pigmentation. This method is innocuous to the subject and easily achieved with a limited understanding of murine anatomy. This refined approach facilitates repeated intubation, which may be necessary for monitoring progression of disease or instillation of treatments. Using this method may result in a reduction of the number of animals and technical skill required to measure lung function in mouse models of respiratory disease.

The goal of this article is to describe a refined method of intubation of the laboratory mouse. The method is noninvasive and, therefore, ideal for studies that require serial monitoring of respiratory function and/or instillation of treatments into the lung.

小鼠重复的奥内切管插管

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ...