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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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?...

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。

<table><tbody><tr><td>SS/45 dumonts</td><td>Fine Science Tools</td><td>11203-25</td><td></td></tr><tr><td>surgical scissors</td><td>Fine Science Tools</td><td>14068-12</td><td></td></tr><tr><td>hemostats</td><td>Fine Science Tools</td><td>13009-12</td><td></td></tr><tr><td>microscissors</td><td>Fine Science Tools</td><td>15000-08</td><td></td></tr><tr><td>0.8 mm curved forceps</td><td>Fine Science Tools</td><td>11009-13</td><td></td></tr><tr><td>suture reel 6-0</td><td>Fine Science Tools</td><td>18020-60</td><td></td></tr><tr><td>suture 4-0 silk w/ needle</td><td>Owens Minor</td><td>K188H</td><td></td></tr><tr><td>gauze 4 x 4</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>cotton-tip applicator</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>30 G needle</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>23 G needle</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>10 cc syringe</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>50 cc conical tube</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>1 cc syringe w/ 25G needle</td><td>Fisher Scientific</td><td>14-826-88</td><td></td></tr><tr><td>Polyethylene 10 tubing 100`(PE-10)</td><td>Fisher Scientific</td><td>14-170-12P</td><td></td></tr><tr><td>Polyethylene 50 tubing 100`(PE-50)</td><td>Fisher Scientific</td><td>14-170-12B</td><td></td></tr><tr><td>3-way stopcock</td><td>Fisher Scientific</td><td>NC9779127</td><td></td></tr><tr><td>surgical blue pad</td><td>Fisher Scientific</td><td>50-7105</td><td></td></tr><tr><td>Sterile Field dressings</td><td>Fisher Scientific</td><td>NC9517505</td><td></td></tr><tr><td>tape rolls 1&quot;</td><td>Corporate Express</td><td>MMM26001</td><td></td></tr><tr><td>straight side wide mouth jars</td><td>VWR</td><td>159000-058</td><td></td></tr><tr><td>stainless steel tray 8&quot; x 11&quot;</td><td>VWR</td><td>62687-049</td><td></td></tr><tr><td>male-male leur lock 3-way</td><td>VWR</td><td>20068-909</td><td></td></tr><tr><td>sterilization pouch 3&quot; x 8&quot;</td><td>VWR</td><td>24008</td><td></td></tr><tr><td>sterilization pouch 5&quot; x 10&quot;</td><td>VWR</td><td>24010</td><td></td></tr><tr><td>absorption triangles</td><td>Fine Science Tools</td><td>18105-03</td><td></td></tr><tr><td>7 mm wound clip applier</td><td>Fisher Scientific</td><td>E0522687</td><td></td></tr><tr><td>1,000 7 mm wound clips</td><td>Fisher Scientific</td><td>E0522687</td><td></td></tr><tr><td>betadine (4 oz)</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>sterile gloves</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>eppendorfs&amp;nbsp;</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>1/2 cc Lo-Dose insulin syringe</td><td>Fisher Scientific</td><td>12-826-79</td><td></td></tr><tr><td>small weigh boat</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>lactated ringers</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>hepranized saline solution (.1 &amp;microL; hep + 9.9 &amp;microL; NaCl)</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>phosphate buffered saline&amp;nbsp;</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>pentobarbital&amp;nbsp;</td><td>can be purchased through any global vendor</td><td></td><td></td></tr><tr><td>Wild M650 microscope w/ boom stand</td><td>Leica</td><td></td><td></td></tr><tr><td>Digi-Med BPA-400 analyzer &amp;amp; systems integrator</td><td>Micro-Med</td><td>SYS-400</td><td></td></tr><tr><td>TXD-310 (Digi-Med Transducer)&amp;nbsp;</td><td>Micro-Med</td><td>TXD-300</td><td></td></tr><tr><td>Computer</td><td>Dell</td><td></td><td></td></tr><tr><td>microbead instrument sterilizer</td><td>VWR</td><td>11156-002</td><td></td></tr><tr><td>Oster A5 clippers w. size 40 blade</td><td>VWR</td><td>10749-020</td><td></td></tr><tr><td>circulating heating pad 18 x 26</td><td>Harvard</td><td>py872-5272</td><td></td></tr><tr><td>rectal thermometer</td><td>Kent Scientific</td><td>RET-3</td><td></td></tr></tbody></table>

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。 <p class="jove_content" fo:keep-tog...

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

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 ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

蛛网膜下腔出血的小鼠模型

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

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. ...

Noninvasive and Invasive Renal Hypoxia Monitoring in a Porcine Model of Hemorrhagic Shock

Up to 50% of patients with trauma develop acute kidney injury (AKI), in part due to poor renal perfusion after severe blood loss. AKI is currently diagnosed based on a change in serum creatinine concentration from baseline or prolonged periods of decreased urine output. Unfortunately, baseline serum creatinine concentration data is unavailable in most patients with trauma, and current estimation methods are inaccurate. In addition, serum creatinine concentration may not change until 24-48 h after the injury. Lastly, oliguria must persist for a minimum of 6 h to diagnose AKI, making it impractical for early diagnosis. AKI diagnostic approaches available today are not useful for predicting risk during the resuscitation of patients with trauma. Studies suggest that urinary partial pressure of oxygen (PuO2) may be useful for assessing renal hypoxia. A monitor that connects the urinary catheter and the urine collection bag was developed to measure PuO2 noninvasively. The device incorporates an optical oxygen sensor that estimates PuO2 based on luminescence quenching principles. In addition, the device measures urinary flow and temperature, the latter to adjust for confounding effects of temperature changes. Urinary flow is measured to compensate for the effects of oxygen ingress during periods of low urine flow. This article describes a porcine model of hemorrhagic shock to study the relationship between noninvasive PuO2, renal hypoxia, and AKI development. A key element of the model is the ultrasound-guided surgical placement in the renal medulla of an oxygen probe, which is based on an unsheathed optical microfiber. PuO2 will also be measured in the bladder and compared to the kidney and noninvasive PuO2 measurements. This model can be used to test PuO2 as an early marker of AKI and assess PuO2 as a resuscitative endpoint after hemorrhage that is indicative of end-organ rather than systemic oxygenation.

Presented here is a protocol to measure renal oxygenation in the medulla and noninvasive urine oxygen partial pressure in a hemorrhagic shock porcine model to establish urine oxygen partial pressure as an early indicator of acute kidney injury (AKI) and a novel resuscitative endpoint.

猪出血性休克模型中的无创和侵入性肾缺氧监测

Up to 50% of patients with trauma develop acute kidney injury (AKI), in part due to poor renal perfusion after severe blood loss. AKI is currently ...

生物工程

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 ...

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 ...

Description of a Swine Infant Model of Volume-Controlled Hemorrhagic Shock

Hemorrhagic shock is a leading cause of morbidity and mortality in pediatric patients. Interpretation of the clinical indicators validated in adults to guide resuscitation and comparison between different therapies is difficult in children due to the inherent heterogeneity of this population. As a result, compared to adults, appropriate management of pediatric hemorrhagic shock is still not well established. In addition, the scarcity of pediatric patients with hemorrhagic shock precludes the development of clinically relevant studies. For this reason, an experimental pediatric animal model is necessary to study the effects of hemorrhage in children as well as their response to different therapies. We present an infant animal model of volume-controlled hemorrhagic shock in anesthetized young pigs. Hemorrhage is induced by withdrawing a previously calculated blood volume, and the pig is subsequently monitored and resuscitated with different therapies. Here, we describe a precise and highly reproducible model of hemorrhagic shock in immature swine. The model yields hemodynamic data that characterizes compensatory mechanisms that are activated in response to severe hemorrhage.

This article aims to provide researchers with a detailed and accessible guide to set up an infant porcine model of hemorrhagic shock.

容量控制性失血性休克的猪婴儿模型的描述

Hemorrhagic shock is a leading cause of morbidity and mortality in pediatric patients. Interpretation of the clinical indicators validated in adults to ...

大鼠血管插管与出血性休克诱导

Developing a Clinically Relevant Hemorrhagic Shock Model in Rats

Over the recent decades, the development of animal models allowed us to better understand various pathologies and identify new treatments. Hemorrhagic shock, i.e., organ failure due to rapid loss of a large volume of blood, is associated with a highly complex pathophysiology involving several pathways. Numerous existing animal models of hemorrhagic shock strive to replicate what happens in humans, but these models have limits in terms of clinical relevance, reproducibility, or standardization. The aim of this study was to refine these models to develop a new model of hemorrhagic shock. Briefly, hemorrhagic shock was induced in male Wistar Han rats (11-13 weeks old) by a controlled exsanguination responsible for a drop in the mean arterial pressure. The next phase of 75 min was to maintain a low mean arterial blood pressure, between 32 mmHg and 38 mmHg, to trigger the pathophysiological pathways of hemorrhagic shock. The final phase of the protocol mimicked patient care with an administration of intravenous fluids, Ringer Lactate solution, to elevate the blood pressure. Lactate and behavioral scores were assessed 16 h after the protocol started, while hemodynamics parameters and plasmatic markers were evaluated 24 h after injury. Twenty-four hours post-hemorrhagic shock induction, the mean arterial and diastolic blood pressure were decreased in the hemorrhagic shock group (p&#160;&#60; 0.05). Heart rate and systolic blood pressure remained unchanged. All organ damage markers were increased with the hemorrhagic shock (p &#60; 0.05). The lactatemia and behavioral scores were increased compared to the sham group (p &#60; 0.05). In conclusion, we demonstrated that the protocol described here is a relevant model of hemorrhagic shock that can be used in subsequent studies, particularly to evaluate the therapeutic potential of new molecules.

作者聚焦:为失血性休克研究开发创新的治疗策略

Hemorrhagic shock kills 1.9 million people worldwide every year. Small animals are frequently used as hemorrhagic shock models but are associated with issues of standardization, reproducibility, and clinical significance, thus limiting their relevance. This article describes developing a new clinically relevant hemorrhagic shock model in rats.

开发大鼠临床相关的出血性休克模型

Over the recent decades, the development of animal models allowed us to better understand various pathologies and identify new treatments. Hemorrhagic ...

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

摘要

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此视频中的章节

固定体积或固定压力：失血性休克的小鼠模型

控制宫颈裂伤损伤小鼠

蛛网膜下腔出血的小鼠模型

全身及肝脏血流动力学参数的监测小鼠

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