该手稿的工作得到了血液医学研究所止血和血管生物学（P3HVB）试验项目计划和AAST研究奖学金资助Neal博士的支持。这项工作得到美国国家卫生研究院授权1 R35 GM119526-01和UM1HL120877-01的支持。

根据匹兹堡大学（美国宾夕法尼亚州匹兹堡）和美国国家卫生研究院（NIH; Bethesda，MD，美国）的动物护理指南，在特定无病原体条件下，12小时光暗循环并自由进入小鼠标准饲料和水。所有动物实验均按照匹兹堡大学动物研究和监督委员会制定的指导方针进行了批准和实施。 手术场和仪器设置<ol><li> 在手术之前，将所有手术器械，缝合线，纱布，棉尖涂抹器，管道和管接头进行灭菌。 <ol><li>在高压灭菌器中灭菌手术器械，缝合线，纱布和棉尖涂抹器。用环氧乙烷灭菌管道和管道连接器。 </li></ol></li><li> 手术领域 <ol><li>打开水循环加热垫并设置在37°C。放手术在其顶部的cal蓝色垫，然后在手术蓝色衬垫顶部的无菌悬垂。 </li><li>将所有灭菌的仪器打开到无菌悬垂物上。使用无菌手套，以避免在此步骤中破坏不育。 </li><li>在不锈钢碗里装上70％乙醇，放在一边。这将用于清洁动物之间的工具。 </li><li>打开微珠灭菌器，加热至150°C。这也将用于清洁动物之间的工具。如果对超过5只小鼠进行手术，请务必将仪器更换为新的无菌组。 </li></ol></li><li> 传感器设置 <ol><li>连接无菌传感器，PE-50管，两个23G针，以及男 - 雄路厄，以及三通旋塞。 6 </li><li>根据制造商的说明书校准并清零传感器。 </li></ol></li></ol>肝脏浸润手术<ol><li> 麻醉诱导和定位 </st荣&gt; <ol><li>以70mg / kg的剂量腹膜内注射戊巴比妥钠。麻醉应在5-10分钟之间生效;用脚趾压缩评估麻醉深度。如果鼠标对脚趾有反应，需要额外的时间或麻醉。如果在手术中需要额外的麻醉，请补充戊巴比妥钠。不要给予大于0.05毫升的补充剂以防止过量。 </li><li>鼠标完全麻醉后，将鼠标仰卧在外科手术板上。用胶带将鼠标四肢固定在板上。 </li><li>用剃刀刮腹部和双侧腹股沟。 </li><li>用betadine浸泡无菌纱布，适用于腹部和双侧腹股沟手术。对于生存实验，用betadine制备腹部和腹股沟，然后用乙醇制备总共三个制备周期。 </li><li>插入直肠温度探头以监控整个过程中的核心温度。保持c矿温在35-37℃之间。 </li></ol></li><li> 股动脉和静脉插管 <ol><li>静脉导管装置:从IV袋中装入PE-10管，30 G针，以及带有乳酸林格氏溶液的三通旋塞。 </li><li>对于动脉导管装置:用肝素化盐水（1,000 U肝素1:10稀释液）填充PE-10管和30G针。需要肝素盐水以防止凝血。 </li><li>将鼠标放在解剖显微镜下。 </li><li>使用手术虹膜剪刀在腹股沟肌肉上进行4-5毫米纵向切口。使用Dumont镊子抓住连接到内收肌的脂肪组织，并横向拉伸以清洁暴露于股骨束。不要通过脂肪组织进行解剖，因为这将导致血管损伤。 </li><li>用Dumont镊子仔细地将神经从动脉和静脉解剖出来。有一个邻近神经的脂肪垫。用一个Dumont强制抓住这个d横向拉这将神经从动脉拉开，创造出​​一条解剖平面。与其他杜蒙镊子直接解剖神经和动脉之间的结缔组织。 <ol><li>在这部分解剖过程中不要抓住神经。 </li></ol></li><li>循环三根6-0丝线缝合在动脉和静脉附近，股骨近端起飞。 <ol><li>将缝合线1放置在近端并放松。 </li><li>将缝合线2放置在最远处，立即绑扎。 </li><li>将缝合线3缝合在缝合线1和2之间，并松开。 </li></ol></li><li>在血管的腹侧表面进行动脉切开术。建议使用微血管剪刀进行动脉切开术以避免血管切断。 </li><li>将三通导管插入动脉。紧接着缝合线1和2将导管固定到位。 </li><li>将三通导管连接到换能器并收集基线血压数据。 </li><li>在对面的腹股沟上重复步骤2.2.4 - 2.2.6。以与动脉相似的方式插入股静脉。在血管的腹侧表面进行静脉切开术，然后插入导管。该导管可用于流体或药物给药。 </li></ol></li><li> 肝脏浸润 <ol><li>预先称重含有0.5mL PBS，三个吸收三角形和每只小鼠一个称重船的管。 </li><li>做一个腹侧中线剖腹手术切口从剑突进程开始，尾部延伸，以使肝脏完全暴露。 </li><li>在腹部插入一个吸收三角形对右侧腹壁。在左侧重复。 <ol><li>确保吸收三角形离开肝脏，以避免肝脏被撕裂后的包装止血作用。 </li></ol></li><li>小心抓住肝左中叶，用外科虹膜剪刀撕裂75％的瓣。放上撕裂d段在含有PBS的管中。 </li><li>用缝钉装置用缝合钉封闭腹壁。一起抓住皮肤和肌肉，然后点燃手指。在肝脏撕裂后尽快做到这一点，以避免腹部外失血。在生存实验中，腹部两层闭合。用于肌肉的可运行的可吸收缝线，随后为肌肤提供不可吸收缝合线的运行层，提供足够的闭合。 </li><li>对于长于30分钟的存活时间点的小鼠，应将股动脉导管取出，将动脉和静脉用缝合线3从步骤2.2.6中取出。然后如上一步骤所述，双侧腹股沟以两层封闭。 </li><li>按照规定的出血时间（30分钟至72小时），取出订书钉。取出吸收三角形并放入相应的称重称重船。使用额外的吸收三角形来吸收任何未吸收的血液。 </li><li&gt;称量吸收过滤器计算总失血量。 </li></ol></li><li> 术后护理 <ol><li>将待处死的小鼠在手术板上30分钟，并在不断监测下，完全麻醉下直至处死。小心安乐死与心脏穿刺和吸入异氟烷过量组合。 </li><li>将处于更长生存时间点的小鼠放置在水循环加热垫顶部的回收笼中。在恢复期间不断监测小鼠，不要离开无人看管，直到他们重新获得维持胸骨躺卧的意识。一旦它从麻醉中恢复，只需将鼠标移到笼子空间。 </li><li>通过皮下注射，每次12小时后，直到呕吐，麻醉后，用0.1mg / kg丁苯替尼治疗术后镇痛。 </li><li>允许老鼠在返回家中后，自由获取食物和水术后正常笼。 </li><li>在牺牲生存小鼠时，用吸入的异氟烷进行麻醉。一旦在麻醉下，通过右心脏心脏穿刺收集血液，如上所述记录失血量，最后用过量的异氟烷保证安乐死。 </li></ol></li></ol>

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作者没有财务竞争的兴趣来宣布。

这里描述的鼠肝切除模型提供了可靠，一致的不受控制的出血模型。这种模式是直接执行的，但是需要仔细考虑的重要步骤。模型中技术上最具挑战性的部分是用于血液动力学监测和流体/药物施用的股骨血管插管。在切除神经和动脉切开术/静脉切开术时必须小心。重要的是在解剖血管期间不要接触神经，以避免造成神经损伤和可能的麻痹，特别是对于生存模型。动脉切开术和静脉切开术需要精细处理血管。我们建议使用微血管剪刀来防止血管意外断断续续。 虽然肝脏撕裂在技术上较困难，但在模型部分中保持一致是重要的，以确保可重复性和一致的出血鼠标。我们的模型旨在在复苏中测试止血剂，因此重要的放置考虑包括确保吸收三角形远离撕裂部位，以避免包装或机械止血作用。避免不必要地操纵其他器官和其他肝叶，以防止在手术过程中无意识的损伤或出血。撕裂后腹部应迅速闭合，以避免腹部外失血。 在整个手术过程中，应仔细监测小鼠，但最重要的是， 如图2所示，给予其经历的显着血液动力学变化。我们对这些显着的血液动力学变化的经验是，如果MAP降低到10mmHg以下&gt; 30秒，则鼠标不太可能存活，我们建议牺牲小鼠URS。如果要测试流体或药物的止血效果，我们建议在撕裂后立即进行治疗，因为小鼠会迅速凝固撕裂区域。如果有兴趣执行比这里描述的更长的观察时间点，则使用疼痛管理至关重要。此外，在股骨血管结扎后，应监测后肢的缺血迹象。由于这些外科手术的丰富经验，本实验室的并发症发生率低于所有测试动物的1％。 4，6 该模型具有许多重要的局限性，包括不受控制的出血的方面。虽然我们看到小鼠血液一致的出血，但是一些小鼠的反应不同，并且在撕裂后会迅速死亡。模型的另一个限制是肝脏撕裂的大小。虽然我们的数据显示了狭窄的标准错误r在切除肝脏的重量中，当由不同个体进行时，切除尺寸的变异性更大，因此出血肯定存在。此外，微血管解剖和插管的学习曲线在技术上可能具有挑战性，我们根据我们的经验估计了50只动物的学习曲线，估计了10只小鼠的学习曲线，描述了肝脏撕裂的再现性。从我们的经验可以预期在72小时56％的存活率。在执行生存分析模型时，密切关注麻醉恢复和适当的疼痛管理至关重要。在我们目前的模型中，我们没有对老鼠进行任何辅助液体或药物复苏，超过了他们在肝切除之前所获得的。重要的是要注意，在模型的生存部分应密切监测动物的窘迫迹象，并适当地治疗疼痛。戊巴比妥是我们的麻醉药对于我们感兴趣的时间点，您可以选择其他选择，但麻醉的其他选择也是可能的，可能会影响结果。疼痛控制对于监测是很重要的，以便小鼠能够自由饮食，如果不能控制，可导致出血和感兴趣的治疗之外的变异性。该模型还适用于其他模型，例如软组织损伤，假骨折或多发性骨折模型。同样，该模型可以容易地适应于研究局部止血剂与静脉注射相比的作用。该模型的多种替代修改是可能的，但仍然未经测试。虽然该模型中的动物与年龄和体重相匹配，但是可以使用不同重量的动物，并且基于体重选择肝脏撕裂的大小。对于可能根据兴趣终点可能不想伤害肝脏的研究者，脾切除术可能预期类似的结果。类似的不控制引导出血模型已经用于其他动物7,8,9,10,11,12，为本模型提供了潜在的替代修改。最后，可以增加撕裂的大小以最大限度地减少血液损失，但是我们已经发现，这大大增加了死亡率，并且扩展模型对主要肝静脉的损伤具有较高的变异程度。 肝脏已被用于以前的不受控制的模型;然而，大多数这些模型是在大鼠模型中进行的。我们在小鼠中发展不受控制的肝脏撕裂模型允许调查人员利用丰富的转基因品种。鼠模型的其他优点包括执行高通量测试，成本效益和易于处理的能力。我们的模型允许血液动力学监测，失血量化和死亡率评估，以前的研究通常只包括评估的这些方面之一。我们能够同时在多个小鼠上执行这个模型，不但允许模型中的高通量但是变化的降低。 总之，我们在这里介绍一种在鼠模型中利用标准肝切除的不受控制的出血的可重复模型。我们的模型是评估新出血或创伤止血药物的理想选择，可用于短期评估失血或进行评估生存效应。

创伤是世界各地年轻人死亡和残疾的主要原因。 1严重受伤创伤患者死亡的主要原因是不受控制的出血。 2出血性创伤患者的管理是双重的:手术出血的控制，血液的复苏和更换。 出血性休克的动物模型一直是创伤研究的基石，可用于评估创伤性/出血性休克的病理生理和治疗。 3，4动物模型的冲击可以通过两种方法实现:控制出血和不受控制的出血。 5，6通过去除固定体积的血液或通过除血实现一定血压（固定压力）进行控制性出血。当时se模型在出血性休克的机制和免疫变化的评估中是有用的，它们不适用于止血剂的测试，并且不模拟创伤后出血的临床情况。在这个程度上，我们试图开发一种不受控制的出血模型，这将允许我们在鼠模型中测试止血变化和促凝剂。肝脏是不受控制的出血的有吸引力的选择，部分原因是肝脏双重供血，而且是钝伤和穿透性创伤中最常见的腹内器官损伤之一。鉴于临床相关性很高，肝脏已被用作不受控制的出血模型，最常见于大鼠和猪模型，但最近也在灵长类动物中。 7，8，9，10，11，12 </su小鼠模型也纳入了肝损伤，如粉碎模型或钝性创伤;然而，这些模型不会导致继发于肝损伤的出血性休克。 13，14 不受控制的肝出血的大鼠和猪模型虽然在复苏实践和血液动力学监测方面有价值，但是由于各种原因（例如成本，使用动物数量）和重要的是相对缺乏可用于分析的转基因品系的特异性细胞和分子信号。目前的鼠模型与现有的肝脏出血模型具有重要的相似之处，包括标准化肝脏撕裂，失血量化，血液动力学监测和进行生存分析的能力。许多现有的模型只包括这些方面的一些，而我们的模型被开发用于测量许多生理变异同时和在多个小鼠。同样，鼠模型的发展打开了复苏之外的调查之门，并在不受控制的出血中形成更大的病理生理机制，具有使用先进分子技术的成本效益高，高通量模型的潜力。

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uncontrolled hemorrhagic shock modeled via liver laceration in mice with real time hemodynamic monitoring

不受控制的出血是创伤患者可预防死亡的重要原因。我们已经通过肝切除开发了一种不受控制的出血的鼠模型，导致持续的失血，血液动力学改变和存活。 小鼠进行肝左中叶的标准化切除。他们被允许出血而没有机械干预。根据研究者的兴趣，止血剂可以作为预处理或拯救治疗来施用。在出血期间，通过左股动脉线进行实时血流动力学监测。然后处死小鼠，量化血液损失，收集血液进行进一步分析，收集器官用于分析损伤。描述了实验设计，以允许同时测试多只动物。 肝出血作为不受控制的出血的模型存在我在文献中，主要在大鼠和猪模型中。这些模型中的一些使用血液动力学监测或量化失血，但缺乏一致性。本模型结合了在鼠模型中的失血定量，实时血液动力学监测，其提供使用转基因品系的优点和高通量机制来进一步研究不受控制的出血的病理生理机制。

肝脏撕裂模型导致小鼠可重复且一致的失血。 图1A示出了可以获得的撕裂肝脏的一致重量，其标准偏差仅为0.02g。撕裂肝脏重量的这种一致性允许在小鼠之间和不同的实验装置中再现模型的能力，例如不同的复苏方案。同样，撕裂肝脏的可重复重量的标准误差仅为0.01g，为动物模型中难以获得的不受控制的出血提供更标准的模型。 验证模型中不同治疗方案的出血效果如图1B所示 。小鼠用肝素预处理（66单位，作为失血的阳性对照）或抗纤维蛋白溶解性，氨甲环酸（TXA）（作为阴性对照），以及先前在鼠尾静脉出血测定中测试的经过验证的止血性纳米颗粒。这些结果证明了该模型用于评估出血设置中止血或抗凝血作用的能力。 不受控制的出血通常伴有监测重要的血液动力学紊乱。在图2中 ，肝切除后各个小鼠的平均动脉血压（MAP）表明，在切开后MAP中的陡峭和可重复的下降导致出血性休克状态。这是重要的，因为它允许不同的复苏或介入措施的血液动力学效应，并允许重要的洞察围绕各种实验条件的生理学。虽然肝脏撕裂后有显着的血液动力学效应，我们发现t该模型可用于评估存活效应，因为该模型已经被评估到72小时，在该时间点存活56％（ 图3 ）。 <img alt="图1" src="/files/ftp_upload/55554/55554fig1.jpg" /> 图1:肝脏浸润验证。 （ A ）横断肝的代表性重量。平均肝脏重量为0.26g，标准偏差为0.02g，平均值为0.01g。这些结果证明了通过我们的75％裂纹的视觉估计可以获得的一致性。 （ B ）用肝素，氨甲环酸和先前验证的止血纳米颗粒预处理后，以克为单位的血液损失平均失血量分别为1.6,0.60和0.65g。这些结果验证了该模型的用途来测试药物的止血或抗凝血作用。 图2:肝脏浸润后平均动脉血压。单个小鼠的图形追踪表示超过20分钟的动脉血迹，其经历了假手术或肝脏撕裂。 肝脏撕裂之后是在假手术小鼠中不存在的小鼠的平均动脉血压（MAP）的特征性和急剧下降。 <img alt="图3" src="/files/ftp_upload/55554/55554fig3.jpg" /> 图3:肝脏浸润后的Kaplan-Meier存活曲线。发现没有任何治疗肝脏撕裂的小鼠的72小时存活率为56％。

Watch this Scientific Journal Video about Uncontrolled Hemorrhagic Shock Modeled via Liver Laceration in Mice with Real Time Hemodynamic Monitoring at JoVE.com

Uncontrolled Hemorrhagic Shock Modeled via Liver Laceration in Mice with Real Time Hemodynamic Monitoring

不可控的出血是创伤患者死亡的重要原因，可以使用鼠模型中的标准肝脏撕裂进行建模。该模型导致失血，存活的一致性，并允许测试止血剂。本文提供了执行此有价值模型的分步过程。

通过实时血液动力学监测在小鼠中通过肝脏浸润建立的不受控制的出血性休克

Uncontrolled hemorrhage is an important cause of preventable deaths among trauma patients. We have developed a murine model of uncontrolled hemorrhage via ...

uncontrolled-hemorrhagic-shock-modeled-via-liver-laceration-mice-with

Nanyang Technological University

Research

JoVE Journal

Medicine

8.5K 次观看.  University of Pittsburgh. 不可控的出血是创伤患者死亡的重要原因，可以使用鼠模型中的标准肝脏撕裂进行建模。该模型导致失血，存活的一致性，并允许测试止血剂。本文提供了执行此有价值模型的分步过程。 

视频: Uncontrolled Hemorrhagic Shock Modeled via Liver Laceration in Mice with Real Time Hemodynamic Monitoring

Fixed Volume or Fixed Pressure: A Murine Model of Hemorrhagic Shock

It is common knowledge that severe blood loss and traumatic injury can lead to a cascade of detrimental signaling events often resulting in mortality. 1, 2, 3, 4, 5 These signaling events can also lead to sepsis and/or multiple organ dysfunction (MOD). 6, 7, 8, 9 It is critical then to investigate the causes of suppressed immune function and detrimental signaling cascades in order to develop more effective ways to help patients who suffer from traumatic injuries. 10 This fixed pressure Hemorrhagic Shock (HS) procedure, although technically challenging, is an excellent resource for investigation of these pathophysiologic conditions. 11, 12, 13 Advances in the assessment of biological systems, i.e. Systems Biology have enabled the scientific community to further understand complex physiologic networks and cellular communication patterns. 14 Hemorrhagic Shock has proven to be a vital tool for unveiling these cellular communication patterns as they relate to immune function. 15, 16, 17, 18 This procedure can be mastered! This procedure can also be used as either a fixed volume or fixed pressure approach. We adapted this technique in the murine model to enhance research in innate and adaptive immune function. 19, 20, 21 Due to their small size HS in mice presents unique challenges. However due to the many available mouse strains, this species represents an unparalleled resource for the study of the biologic responses. The HS model is an important model for studying cellular communication patterns and the responses of systems such as hormonal and inflammatory mediator systems, and danger signals, i.e. DAMP and PAMP upregulation as it elicits distinct responses that differ from other forms of shock. 22, 23, 24, 25 The development of transgenic murine strains and the induction of biologic agents to inhibit specific signaling have presented valuable opportunities to further elucidate our understanding of the up and down regulation of signal transduction after severe blood loss, i.e. HS and trauma 26, 27, 28, 29, 30. 
There are numerous resuscitation methods (R) in association with HS and trauma. 31, 32, 33, 34 A fixed volume resuscitation method of solely lactated ringer solution (LR), equal to three times the shed blood volume, is used in this model to study endogenous mechanisms such as remote organ injury and systemic inflammation. 35, 36, 38 This method of resuscitation is proven to be effective in evaluating the effects of HS and trauma 38, 39.

The Hemorrhagic Shock model has been a reliable and reproducible resource facilitating the identification and understanding of signaling cascades associated with inflammation and end-organ damage after trauma. This article provides a step-by-step description of surgical and mechanical aspects associated with the Hemorrhagic Shock experimental procedure in mice.

固定体积或固定压力:失血性休克的小鼠模型

It is common knowledge that severe blood loss and traumatic injury can lead to a cascade of detrimental signaling events often resulting in mortality. 1, ...

科研

医学

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

控制宫颈裂伤损伤小鼠

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

Real-time Cytotoxicity Assays in Human Whole Blood

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with high-throughput cell positioning technology. The targeted tumor cell populations are first preprogrammed to immobilization into an array format, and labeled with green fluorescent cytosolic dyes. Following the cell array formation, antibody drugs are added in combination with human whole blood. Propidium iodide (PI) is then added to assess cell death. The cell array is analyzed with an automatic imaging system. While cytosolic dye labels the targeted tumor cell populations, PI labels the dead tumor cell populations. Thus, the percentage of target cancer cell killing can be quantified by calculating the number of surviving targeted cells to the number of dead targeted cells. With this method, researchers are able to access time-dependent and dose-dependent cell cytotoxicity information. Remarkably, no hazardous radiochemicals are used. The WCA presented here has been tested with lymphoma, leukemia, and solid tumor cell lines. Therefore, WCA allows researchers to assess drug efficacy in a highly relevant ex&#160;vivo condition.

The whole blood cytotoxicity assay (WCA) is a cytotoxicity assay developed by incorporating high-throughput cell positioning technology with fluorescence microscopy and automated image processing. Here, we describe how lymphoma cells treated with an anti-CD20 antibody can be analyzed real-time in human whole blood to provide quantitative cellular cytotoxicity analysis.

实时的细胞毒性分析在人全血

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...

Monitoring of Systemic and Hepatic Hemodynamic Parameters in Mice

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. However, due to the small size of the mouse, technical details of the intraoperative monitoring procedure suitable for the mouse were rarely described. Previously we have reported a monitoring procedure to obtain hemodynamic parameters for rats. Now, we adapted the procedure to acquire systemic and hepatic hemodynamic parameters in mice, a species ten-fold smaller than rats. This film demonstrates the instrumentation of the animals as well as the data acquisition process needed to assess systemic and hepatic hemodynamics in mice. Vital parameters, including body temperature, respiratory&#160;rate&#160;and heart&#160;rate were recorded throughout the whole procedure. Systemic hemodynamic parameters consist of carotid artery pressure (CAP) and central venous pressure (CVP). Hepatic perfusion parameters include portal vein pressure (PVP), portal flow rate as well as the flow rate of the common hepatic artery (table 1). Instrumentation and data acquisition to record the normal values was completed within 1.5 h. Systemic and hepatic hemodynamic parameters remained within normal ranges during this procedure.
This procedure is challenging but feasible. We have already applied this procedure to assess hepatic hemodynamics in normal mice as well as during 70% partial hepatectomy and in liver lobe clamping experiments. Mean PVP after resection (n= 20), was 11.41&#177;2.94 cmH2O which was significantly higher (P&#60;0.05) than before resection (6.87&#177;2.39 cmH2O). The results of liver lobe clamping experiment indicated that this monitoring procedure is sensitive and suitable for detecting small changes in portal pressure and portal flow rate. In conclusion, this procedure is reliable in the hands of an experienced micro-surgeon but should be limited to experiments where mice are absolutely needed.

This film demonstrates how to acquire systemic and hepatic hemodynamics in mice. The whole monitoring includes acquisition of vital parameters, systemic blood pressure, central venous pressure, common hepatic artery flow rate, and portal vein pressure as well as the portal flow rate in mice.

全身及肝脏血流动力学参数的监测小鼠

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. ...

Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any potential therapeutic approach directed against acute left ventricular (LV) dysfunction. Current imaging modalities (e.g., echocardiography and magnetic resonance imaging) have several limitations since data on LV pressure cannot directly be measured. LV catheterization in mice undergoing coronary artery occlusion could serve as a novel method for a real-time evaluation of LV function.
At the beginning of the procedure, mice were anesthetized followed by endotracheal intubation. For LV catheterization, the right carotid artery was exposed via middle-neck incision. The catheter was introduced and placed into the LV cavity. Left thoracotomy was conducted and the left main coronary artery (LCA) was ligated. To induce reperfusion, the suture was released after 45 min. Pressure-volume data was recorded at all times.
Ligation of the LCA caused a decrease in LV systolic function as evidenced by a 30% reduction in stroke volume, LV ejection fraction (EF), and cardiac output. Maximum dP/dt as a parameter for LV contractility was also significantly reduced and diastolic function was severely impaired (minimum dP/dt -40%). Reperfusion over a period of 20 min did not lead to a complete recovery of LV function.
Real-time pressure-volume analysis served as a valid procedure for monitoring cardiac function during acute myocardial infarction in mice. Maintaining stable anesthesia and a standardized surgical approach was crucial to ensure valid results. As the early phase of acute myocardial infarction is critical for morbidity and mortality, the delineated method could be beneficial for preclinical evaluation of new strategies for cardioprotection.

Acute myocardial infarction in mice induces acute but incompletely characterized changes in left ventricular (LV) function. LV catheterization in mice undergoing coronary artery occlusion serves as a novel method for a real-time evaluation of LV function.

小鼠急性心肌梗死的实时压力容积分析

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any ...

Complete and Partial Aortic Occlusion for the Treatment of Hemorrhagic Shock in Swine

Hemorrhage remains the leading cause of preventable deaths in trauma. Endovascular management of non-compressible torso hemorrhage has been at the forefront of trauma care in recent years. Since complete aortic occlusion presents serious concerns, the concept of partial aortic occlusion has gained a growing attention. Here, we present a large animal model of hemorrhagic shock to investigate the effects of a novel partial aortic balloon occlusion catheter and compare it with a catheter that works on the principles of complete aortic occlusion. Swine are anesthetized and instrumented in order to conduct controlled fixed-volume hemorrhage, and hemodynamic and physiological parameters are monitored. Following hemorrhage, aortic balloon occlusion catheters are inserted and inflated in the supraceliac aorta for 60 min, during which the animals receive whole-blood resuscitation as 20% of the total blood volume (TBV). Following balloon deflation, the animals are monitored in a critical care setting for 4 h, during which they receive fluid resuscitation and vasopressors as needed. The partial aortic balloon occlusion demonstrated improved distal mean arterial pressures (MAPs) during the balloon inflation, decreased markers of ischemia, and decreased fluid resuscitation and vasopressor use. As swine physiology and homeostatic responses following hemorrhage have been well-documented and are like those in humans, a swine hemorrhagic shock model can be used to test various treatment strategies. In addition to treating hemorrhage, aortic balloon occlusion catheters have become popular for their role in cardiac arrest, cardiac and vascular surgery, and other high-risk elective surgical procedures.

Here, we present a protocol demonstrating a hemorrhagic shock model in swine that uses aortic occlusion as a bridge to definitive care in trauma. This model has application in testing a wide range of surgical and pharmacological therapeutic strategies.

完全性和部分主动脉闭塞治疗猪失血性休克

Hemorrhage remains the leading cause of preventable deaths in trauma. Endovascular management of non-compressible torso hemorrhage has been at the ...

Measurement of the Hepatic Venous Pressure Gradient and Transjugular Liver Biopsy

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced chronic liver disease followed by an instruction for transjugular biopsy. Under local anesthesia and ultrasound guidance, a catheter introducer sheath is placed in the right internal jugular vein. Using&#160;fluoroscopic guidance, a balloon catheter is advanced into the inferior vena cava (IVC) and inserted into a large hepatic vein. Correct and sufficient wedge position of the catheter is ensured by injecting contrast media while the balloon is blocking the outflow of the cannulated hepatic vein. After calibrating the external pressure transducer, continuous pressure recordings are obtained with triplicate recordings of the wedged hepatic venous pressure (WHVP) and free hepatic venous pressure (FHVP). The difference between FHVP and WHVP is referred to as HVPG, with values &#8805;10 mm Hg indicating clinically significant portal hypertension (CSPH). Before removing the catheter, pressure readings obtained in the IVC at the same level, as well as the right atrial pressure are recorded.
Finally, a transjugular liver biopsy can be obtained via the same vascular route. Different systems are available; however, core biopsy needles are preferred over aspiration needles, especially for cirrhotic livers. Again, under fluoroscopic guidance a biopsy needle introducer sheath is advanced into an hepatic vein. Next, the transjugular biopsy needle is gently advanced through the introducer sheath: (i) in case of aspiration biopsy, the needle is advanced into the liver parenchyma under aspiration and then removed quickly, or (ii) in case of a core biopsy, the cutting-mechanism is triggered inside the parenchyma. Several separate passages can be safely performed to obtain sufficient liver specimens via transjugular biopsy. In experienced hands, the combination of these procedures takes about 30-45 min.

Here, we present a protocol for measurement of hepatic venous pressure gradient (HVPG),the gold standard to diagnose clinically significant portal hypertension. Moreover, we describe how to perform a transjugular liver biopsy within the same session.

肝静脉压梯度和转性肝活检测量

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced ...

Standardized Hemorrhagic Shock Induction Guided by Cerebral Oximetry and Extended Hemodynamic Monitoring in Pigs

Hemorrhagic shock ranks among the main reasons for severe injury-related death. The loss of circulatory volume and oxygen carriers can lead to an insufficient oxygen supply and irreversible organ failure. The brain exerts only limited compensation capacities and is particularly at high risk of severe hypoxic damage.This article demonstrates the reproducible induction of life-threatening hemorrhagic shock in a porcine model by means of calculated blood withdrawal. We titrate shock induction guided by near-infrared spectroscopy&#160;and extended hemodynamic monitoring to display systemic circulatory failure, as well as cerebral microcirculatory oxygen depletion. In comparison to similar models that primarily focus on predefined removal volumes for shock induction, this approach highlights a titration by means of the resulting failure of macro- and microcirculation.

Hemorrhagic shock is a severe complication in seriously injured patients, which leads to life-threatening oxygen undersupply. We present a standardized method to induce hemorrhagic shock via blood withdrawal in pigs that is guided by hemodynamics and microcirculatory cerebral oxygenation.

由脑氧测量和猪期扩展血液动力学监测指导的标准化出血性休克诱导

Hemorrhagic shock ranks among the main reasons for severe injury-related death. The loss of circulatory volume and oxygen carriers can lead to an ...

Real-Time Monitoring of Neurocritical Patients with Diffuse Optical Spectroscopies

Neurophysiological monitoring is an important goal in the treatment of neurocritical patients, as it may prevent secondary damage and directly impact morbidity and mortality rates. However, there is currently a lack of suitable non-invasive, real-time technologies for continuous monitoring of cerebral physiology at the bedside. Diffuse optical techniques have been proposed as a potential tool for bedside measurements of cerebral blood flow and cerebral oxygenation in case of neurocritical patients. Diffuse optical spectroscopies have been previously explored to monitor patients in several clinical scenarios ranging from neonatal monitoring to cerebrovascular interventions in adults. However, the feasibility of the technique to aid clinicians by providing real-time information at the bedside remains largely unaddressed. Here, we report the translation of a diffuse optical system for continuous real-time monitoring of cerebral blood flow, cerebral oxygenation, and cerebral oxygen metabolism during intensive care. The real-time feature of the instrument could enable treatment strategies based on patient-specific cerebral physiology rather than relying on surrogate metrics, such as arterial blood pressure. By providing real-time information on the cerebral circulation at different time scales with relatively cheap and portable instrumentation, this approach may be especially useful in low-budget hospitals, in remote areas and for&#160;monitoring in open fields (e.g., defense and sports).

Presented here is a protocol for non-invasively monitoring cerebral hemodynamics of neurocritical patients in real-time and at the bedside using diffuse optics. Specifically, the proposed protocol uses a hybrid diffuse optical systems to detect and display real-time information on cerebral oxygenation, cerebral blood flow and cerebral metabolism.

使用漫反射光谱实时监测神经危重症患者

Neurophysiological monitoring is an important goal in the treatment of neurocritical patients, as it may prevent secondary damage and directly impact ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

器官缺血-输液损伤模拟大鼠肝脏移植模型的血动力学变化

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...