我们想感谢雷切尔 O ' 康奈尔, 和杰西卡李为他们的帮助, 动物研究。我们也要承认少校哈罗德 Timboe, MD, 英里, 美国陆军 (Ret), 谁是这个项目的顾问和导师。

在使用动物进行研究时, 调查人员遵守了《动物福利法条例》和其他有关动物和实验的联邦法规以及有关动物的试验和《护理和使用指南》现行版本中规定的原则。国家研究委员会的实验动物。这项研究议定书获得了密歇根大学机构动物保育和使用委员会 (IACUC) 的批准。实验是按照所有有关动物福利的条例和准则进行的。
1. 动物的选择和驯化
<ol><li>使用女约克郡-长白杂交猪 (n = 2/组, 年龄范围: 5-6 月, 重量范围:40-60 公斤) 为实验。</li>
	<li>他们到达该设施后, 将动物单独安置在笼子里, 但要确保相邻笼子里的动物之间有充分的互动。</li>
	<li>将动物饲养至少5维, 以便在进行实验之前完全适应。在此期间, 喂养动物的标准饮食, 并观察他们, 以确定他们处于良好的健康状态。</li>
	<li>在手术前午夜防止动物进食和饮用, 以防插管时有吸入的危险。</li>
</ol>2. 麻醉和插管
<ol><li>用5毫克/千克 tiletamine/zolazepam 给予肌肉 (IM) 进入臀肌, 而动物则是在房屋区, 使动物镇静。同时, 给予0.05 毫克/千克阿托品 IM 作为抗胆碱能剂, 以尽量减少在插管期间的分泌物。</li>
	<li>将动物从房屋区运送到手术室, 并将其放置在手术台上的仰卧位。</li>
	<li>使用吸入异氟醚 (2%-5%) 诱发麻醉。使用21% 氧气 (0.4 升/分) 混合空气 (4 升/分钟) 作为载体气体, 以提供吸入麻醉气体。</li>
	<li>在达到所需的麻醉深度的同时, 将20克外周静脉 (iv) 导管放置在耳静脉内, 用于静脉注射液体、药物或麻醉, 如有需要。</li>
	<li>当动物获得所需的麻醉深度时, 轻轻地冲洗掉手术部位的任何废物和碎片, 包括左、右腹股沟区域、颈部左右两侧和腹部。使用电动理发剪去除所有的头发从手术现场。通过捏紧动物的后肢并观察其 (缺乏) 反应来评估麻醉深度。</li>
	<li>在气管插管前, 用手术棉绳将动物安全地放到手术台上。</li>
	<li>用一根短的手术棉绳抬起下颌骨, 用舌头打开口腔口。用另一根棉线把上颌向下拉。如果下颚松弛不足或声带折叠, 则应简要增加异氟醚的水平, 并重复此步骤。</li>
	<li>使用非占主导地位的手持有传统喉镜安装与12在点燃的米勒刀片手柄。通过咽的米勒刀片的尖端。在口腔内机动刀片时, 应注意保持牙齿和口腔粘膜。缓慢前进的刀片提示, 直到会厌可以清楚地可视化, 并提前通过喉入口。用手腕, 抬起刀刃, 抬高会厌, 这样就能清楚地看到喉部。</li>
	<li>放置一个6.5 法郎或7法郎气管内管 (ETT) 与鞘之间的声带到气管。在喉痉挛的情况下, 应用2% 利多卡因在声带和中位 glosso-会厌领域, 重新评估。</li>
	<li>取出鞘, 然后将管子完全推进气管向下, 同时保持牢牢地抓住管子。充气气球袖口与 10-15 厘米3的空气, 以防止任何空气泄漏周围的袖口和愿望的胃内容。</li>
	<li>通过呼吸过滤器 (热和湿式换热器) 将 ETT 连接到机械通风机。通过把棉胶带绑在鼻子上 (上颌骨) 来保护 ETT。</li>
	<li>在一次失败的插管的情况下, 让动物氧和透气简要地在任何其他企图之前。</li>
	<li>确保适当的机械通风机设置: 混合21% 氧气和空气;7-10 毫升/千克体重的潮汐体积;呼吸率为 10-15 呼吸/分钟, 以维持一个最终潮汐的病,2的 40 mmHg @ 2 mmHg。</li>
	<li>将脉冲脉搏在耳朵、舌头或尾部, 以测量血液的氧合和心率。通过肛门口推进直肠探头进行连续温度监测。 注意: 动物的温度将被维护在38°c 之间到40°c 使用被加热的温度被调控的操作桌、热垫或热化毯子。</li>
	<li>使用异氟醚 (1%-3%) 在整个过程中保持麻醉。在这段时间, 评估麻醉深度, 通过测试疼痛反射使用后肢捏。此外, 还要监测呼吸机上的呼吸系统, 看看是否符合程序的呼吸机率。 注: 过度呼吸表明麻醉深度不足。</li>
</ol>3. 外科现场灭菌 (准备和悬垂)
<ol><li>准备切口部位, 以及周围的广泛区域为实验程序。该试验的切口部位包括双侧腹股沟区、双侧颈和下腹部。对手术部位进行消毒, 用大量的聚维酮碘5分钟擦洗。</li>
	<li>使用干燥, 无菌纱布从皮肤去除聚维酮碘肥皂。</li>
	<li>在手术部位周围放置无菌的手术巾, 以保存无菌手术场。使用毛巾钳固定毛巾到位。用不育的褶皱覆盖动物, 防止手术部位受到任何污染。</li>
</ol>4. 插管
<ol><li>
股动脉和静脉插管 注: 对右股动脉、左股动脉和左股静脉进行插管。右股动脉是空心使用14法郎插入鞘的主动脉球囊闭塞导管插入。左股动脉是空心的, 使用5法郎导管进行远端地图监测。在左股静脉内放置8法郎导管, 用于体液管理和血液提取。开放 Seldinger 技术用于获得所有插管15的血管通路。<ol>
<li>在做切口之前, 用捏下动物的后肢来评估麻醉的深度。</li>
		<li>使用10号无菌手术刀片的手术刀, 在右腹股沟4厘米以上和右腹股沟折痕下方4厘米的垂直8厘米切口进行。</li>
		<li>解剖通过皮下组织和肌肉, 并使用两个 Weitlaner 拉钩获得适当的接触。如果需要, 使用陆军海军牵引器进行额外的收缩。</li>
		<li>使用 Mixter 直角钳和烙解剖通过结缔组织, 直到神经血管束清楚暴露。</li>
		<li>小心解剖动脉。保持神经, 这是最侧面的结构。</li>
		<li>仔细解剖静脉, 它位于最内侧, 远离动脉。</li>
		<li>用2-0 丝领带获得动脉近端和远端控制。</li>
		<li>用20克角介绍人针穿刺动脉。通过观察针的另一端的脉动血流, 确保血管腔内的适当位置。</li>
		<li>通过斜针的流明, 在导丝上推进一个圆尖的0.35。</li>
		<li>把针从导丝上取出。确保导丝的迁移不被保存到位。</li>
		<li>通过一个10法郎扩张在导丝扩大在动脉的开放。卸下10法郎扩张。在导丝上插入14法郎插入鞘。</li>
		<li>小心地将扩张从插入鞘与导丝一起取出, 保持动脉内的套管。</li>
		<li>冲洗插入鞘以确保其在血管腔内的位置。</li>
		<li>将导管的末端固定在 3-0 polyglactin 缝合处。</li>
		<li>使用3-0 尼龙缝合, 以执行临时运行关闭的覆盖皮肤。</li>
		<li>重复上述步骤的左股动脉插管使用5法郎导管 (没有初始扩张)。以类似的方式隔离左股静脉, 用 8 cannulate 导管将其分离。使用3-0 尼龙缝合, 以执行临时运行关闭的覆盖皮肤。</li>
	</ol>
</li>
	<li>
颈动脉和颈外静脉插管 注: 进行双侧颈外静脉插管和左颈总动脉穿刺。5法郎导管放置在左颈总动脉中, 用于近端图监测, 左颈外静脉8法郎导管用于液体管理和升压输液的其他中心静脉通路。9法郎导管放置在右颈外静脉肺动脉插管肺动脉导管 (如天鹅甘兹) 导管, 而颈动脉血流探头放置在右颈总动脉周围的颈动脉血流率监测。Seldinger 方法用于获得所有插管的血管通路。<ol>
<li>使用10刀片手术刀, 使6厘米垂直切口约2厘米侧向中线在颈部左侧。</li>
		<li>使用烙解剖通过皮下组织直到胸锁乳突 (SCM) 肌肉暴露。</li>
		<li>将 Weitlaner 拉钩放在切口的近端和远端, 以完全收缩。</li>
		<li>沿单片肌的侧缘解剖, 露出左颈外静脉。</li>
		<li>通过以下步骤 4.1.8 4.1.14 可以实现导管插管。插入, 冲洗, 并确保静脉8法郎导管。</li>
		<li>对于左颈总动脉暴露, 解剖单片机肌肉的内侧边缘。放置一个 Weitlaner 的牵引器, 以增强曝光。</li>
		<li>沿气管的外侧边界纵向解剖结缔组织。如果遇到胸腺腺体, 请保存它们。</li>
		<li>暴露颈动脉三角形, 其中包括颈动脉粥样硬化, 颈内静脉和迷走神经。触及颈动脉以确定其弹道。</li>
		<li>仔细解剖颈总动脉远离静脉和神经。</li>
		<li>通过以下步骤 4.1.8 4.1.14 执行颈动脉插管。插入, 冲洗, 并确保在动脉5法郎导管, 如前所述。</li>
		<li>重复步骤4.1。对右颈外静脉和右颈部总动脉的解剖和分离。</li>
		<li>在右颈总动脉周围放置4毫米颈动脉血流探头。在流探针和容器之间应用换能器凝胶, 以获得最佳的流量信号和捕获。</li>
		<li>按照步骤 4.1. 8-4. 1.14, cannulate 右颈外静脉使用9法郎介绍人鞘。冲洗并固定导管到位。使用3-0 尼龙缝合, 以执行临时运行关闭的覆盖皮肤。</li>
	</ol>
</li>
</ol>5. PA 导管插入
<ol><li>用生理盐水 (NS) 冲洗注射口、近端口和导管远端, 并将其连接到换能器油管。远端端口被指定为肺动脉 (PA) 端口, 而近端端口被指定为中心静脉压力 (CVP) 端口。</li>
	<li>通过移动导管检查显示器上的工件跟踪。这样做是为了确保导管能正常工作。</li>
	<li>通过无菌套筒推进 PA 导管。</li>
	<li>使用3厘米3注射器, 充气的 PA 导管气球, 不超过1.5 厘米3的空气, 以测试通货膨胀。使气球充气, 将导管插入鞘内。</li>
	<li>插入 PA 导管通过9法郎插入鞘。一旦 PA 导管插入至少18厘米, 充气气球不超过1.5 厘米3的空气。</li>
	<li>在插入过程中, 缓慢地推进 PA 导管并检查显示器是否有心律失常。在心室异位的情况下, 充气和放气气球以防止进展。如果插入变得困难以任何方式, 气球应该是充气的, 并且导管应该被收回和重新插入。</li>
	<li>观察显示器以评估从右心房 (RA) 到右心室 (RV) 向 PA 到肺动脉楔压的转变。</li>
	<li>使气球充气, 并确认 PA 跟踪返回到显示器。 注: 右颈外静脉插入部位导管的适当长度约为 45-55 厘米。</li>
	<li>将无菌套筒的末端连接到插入鞘, 以确保导管在 PA 内的位置。</li>
	<li>将 PA 导管的插入端口连接到捕获单元, 以获得心脏输出 (CO) 和静脉氧饱和度 (SVO2)。</li>
	<li>校准 PA 导管监控系统。利用动物的身体长度和重量, 对动物的 PA 导管监测系统进行体内标定。使用静脉血气进一步校准使用氧饱和度和血红蛋白水平。</li>
</ol>6. 膀胱管放置
<ol><li>用10刀片手术刀在中线做一个5厘米下腹切口。</li>
	<li>使用烙, 解剖通过皮下组织和 linea。在解剖过程中可视化每一层。</li>
	<li>Extracorporealize 膀胱。</li>
	<li>用两个迈克迪贝克钳, 抓住腹壁上的膀胱, 远离输尿管开口。</li>
	<li>使用烙, 在膀胱内做一个小开口, 露出内腔。</li>
	<li>使用吸入去除膀胱腔内的任何尿液。手术海绵可用于任何意外溢出尿在膀胱外。</li>
	<li>使用4-0 聚丙烯缝合, 以执行临时的钱包字符串关闭膀胱。</li>
	<li>在膀胱腔内放置一个18法郎的导尿管, 并使用10厘米3注射器充气气球。将尿引流袋连接到导尿管, 并将其固定在手术台的一侧。</li>
	<li>配合4-0 聚丙烯缝合, 以确保导尿管在膀胱腔内。</li>
	<li>使用3-0 尼龙缝合, 以执行临时运行关闭的腹部上覆皮肤。</li>
</ol>7. 完全和部分主动脉球囊导管插入术
<ol><li>随机的动物接受完全或部分主动脉球囊闭塞导管。</li>
	<li>插入0.035 英寸, 260 厘米血栓消融僵硬导丝通过14法郎插入鞘在右股动脉, 并引导它进入 supraceliac 主动脉 (1 区) 使用超声检查腹部和胸主动脉。</li>
	<li>将气囊闭塞导管插入导丝的1区, 在出血后进行闭塞。</li>
	<li>通过超声检查确定主动脉球囊闭塞导管进入主动脉1区的最终位置。</li>
</ol>8. 术中血流动力学和实验室监测
<ol><li>确保侵入导管与监测系统的连接, 使用油管、传感器和导管进行血流动力学监测。侵入性监测系统应在动物心脏的水平上 "归零", 以确保准确的监测。</li>
	<li>在整个实验中记录生理数据, 包括近端和远端图、心率 (HR)、CVP、CO、PA 压力、SVO2、终潮汐 CO2和核心体温。</li>
	<li>每5分钟在出血和气球的通货膨胀阶段记录这些变量, 在余下的实验中每15分钟。</li>
	<li>收集血液样本的4个时间点: 基线 (BL), 后休克 (PS), 复苏后 (PR), 并在结束 (E) 的实验。在每一个时间点为血浆和血清分析收集10毫升的血液。</li>
	<li>通过收集多达1毫升的血液在 ABG 注射器进行动脉血气 (ABG) 取样。在上述每个时间点执行 ABG 取样, 并且在实验中需要时进行。使用血气分析仪进行分析。 注: 每样样品应在抽取血液的10分钟内进行分析, 因为延迟时间超过10分钟会降低结果16的效率。</li>
</ol>9. 出血
<ol><li>计算 TBV。 TBV 在 mL =重量在 g x 0.06 + 0.77</li>
	<li>出血35% 的 TBV 使用一个自动泵在20分钟的时间内删除一半以上的前7分钟, 另一半在未来13分钟。</li>
	<li>收集血液在标准采血袋。储存在4摄氏度, 预计未来输血。</li>
	<li>如果地图减少低于 30 mmHg, 停止出血, 关闭异氟醚, 并管理 50-100 毫升 boluses 的 NS。</li>
	<li>当地图返回到 30 mmHg 时, 重新启动出血。</li>
</ol>10. 主动脉球囊闭塞导管充气和全血复苏
<ol><li>充气主动脉球囊闭塞导管与 9-12 厘米3的空气或直到没有进一步下降, 在远端地图后, 额外的气球通货膨胀被注意到。</li>
	<li>从部分主动脉球囊闭塞导管中取出导线, 以促进远端主动脉流, 同时将其留作完全主动脉球囊闭塞导管。通过超声检查确定动脉球囊闭塞导管在1区的位置, 排除导管移植。</li>
	<li>使用中断的3-0 尼龙缝合将导管固定在皮肤上。</li>
	<li>在40分钟的主动脉闭塞后, 通过使用自动泵, 在20分钟内通过左股静脉导管, 将全血量相等的动物恢复到 TBV 的20%。使用血液温暖的输血, 以40摄氏度的温度为目标。</li>
	<li>复苏后, 气球的充气量递增5分钟。</li>
	<li>从14法郎插入鞘中取出主动脉球囊闭塞导管。</li>
	<li>鉴于已知的缺血再灌注损伤的危险与相关的低血压, 在这段时间密切监测血流动力学。</li>
</ol>11. 关键护理、观察和恢复
<ol><li>观察动物4小时, 持续监测其生理和实验室参数。</li>
	<li>提供维持静脉输液 (NS 灌注为50毫升/小时)。</li>
	<li>以 6 mmHg 为目标, 通过管理30毫升/千克 boluses 的 NS, 其次是20毫升/千克 boluses 当 cvp 低于目标。每小时评估流体反应性。</li>
	<li>使用去甲肾上腺素 (0.024 毫克/毫升) 的 60 mmHg 地图。</li>
	<li>使用加热毯保持 38-40 摄氏度的生理温度。</li>
	<li>在4小时的结束, 弄死的动物与戊巴比妥钠 (100 毫克/千克) 注射。</li>
</ol>12. 尸检
<ol><li>从动物身上取出所有必需的线和管子。</li>
	<li>准备一个容器与常规冰, 并放置在操作表旁边, 以冻结器官组织后立即从身体切除。</li>
	<li>使用手术刀做一个6厘米长的垂直切口沿左胸骨边界, 解剖通过皮肤, 皮下组织, 胸肌和肋软骨。进入胸腔, 露出心脏和肺部。</li>
	<li>用手术刀把心包膜切开, 用镊子抓住心脏的顶端。用手术刀切开心脏的 5 x 5 cm 部分在顶点。</li>
	<li>用剪刀把肺的 5 x 5 厘米的部分从肺底部的前边界切开。</li>
	<li>将开胸切口延伸到腹部前中线, 露出腹部内脏。</li>
	<li>使用钳持有肝脏的左叶和削减 5 x 5 厘米部分。</li>
	<li>重复相同的过程, 从脾脏获得样品。</li>
	<li>用右手创建一个窗口进入左腹腔抓住肾脏。肾实质被动员后, 横断面肾动脉、静脉和输尿管。取出肾囊。</li>
	<li>轻轻挤压大便远离5厘米长的小肠, 用剪刀从肠系膜上切开5厘米长的肠道。</li>
	<li>用手术刀从大腿上切开 5 x 2 厘米长的四头肌肌肉。</li>
	<li>通过将这些样品切割成离心管中的小段来进一步处理。通过将管子放置在干冰或液氮中, 将这些组织样品进行闪光冷冻。</li>
	<li>使用50毫升圆锥管, 含有福尔马林溶液, 固定3厘米长, 细切切片组织进行组织学评估。</li>
</ol>

<ol>
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这项工作得到美国陆军医学研究和物资司令部的支持。W81XWH-16-C-0102。本报告所载的意见、意见和/或调查结果均为提交人, 不应被解释为军队立场、政策或决定的正式部门, 除非由其他文件指定。

在本议定书中, 我们强调了猪失血性休克模型。该模型已被证明是可靠的和可重现的16,17,18,19。类似于此的模型已被用于研究失血性休克对动物生理学16,20的影响的一些科学研究。此外, 该模型还用于测试药物和手术治疗的出血性休克的干预, 标志着成功12,13,16,19, 21。
这个模型包含了需要非常注意细节的几个步骤。猪的插管是一个复杂的过程, 因为动物有一个长, 喙状的鼻子和狭窄, 长口咽腔。此外, 猪一般有很高的倾向, 接受喉痉挛, 使气管插管更具挑战性的22。适当的诱导麻醉, 促进良好的肌肉松弛, 应在尝试插管前达到。根据我们的经验, 有一个助手使用外科棉绳抬起下颌和舌头, 并压低上颌有助于促进更广泛的口腔开放, 以方便插管。如果插管变得困难由于喉痉挛, 应使用局部利多卡因喷雾。虽然没有在本议定书中使用, 一个神经肌肉阻滞剂可用于促进喉松弛。然而, 使用神经肌肉阻滞剂需要训练有素的专业人员进行严密的监督。我们还发现将润滑凝胶应用于气管导管的末端, 以及在通过喉开口推进导管的同时旋转管是有帮助的。气管插管后, 应用末端潮汐 CO2确认插管。然而, 虽然猪可以在不同的位置插管, 我们发现在仰卧位的插管是最容易的, 特别是如果个人执行插管有经验的人气管插管。
股动脉和静脉插管在技术上是有挑战性的。使用良好的回缩是很重要的, 可以通过使用自保持拉钩来实现。如果需要, 可以使用额外的牵引器, 如陆军海军。在解剖神经血管束时应注意, 因为股神经是捆绑中最侧的结构, 必须保留。这是特别重要的, 如果动物需要生存的实验。另外, 右股动脉插管是实验的关键步骤。在导丝插管进入容器后, 插入鞘14法郎。成功执行这一步骤需要一个初始扩张与10法郎扩张, 以升迁船只。同样重要的是压缩 arteriotomy 部位的股动脉, 除去10法郎扩张后, 以最小化失血。虽然在动物研究中不经常进行, 但在执行 arteriotomy 和 venotomy 之前获得近端和远端控制, 如本研究所示, 可以帮助减少出血, 并允许进行故障排除, 如果出现问题时,插管。
正确定位和部署主动脉球囊闭塞导管是至关重要的。在推进导管内的主动脉中应谨慎, 因为攻击性的操作可能导致对股动脉或主动脉的医源性损伤。虽然有几个地点可能是部署导管的目标, 我们选择的位置的阻断气球在主动脉区 1, 这是 supraceliac 主动脉。气球定位可以通过手动触诊或透视来确认;然而, 超声也可以用来方便地确认气球放置, 这是用于本研究。在适当的定位后, 气球的通胀应该小心进行。一般来说, 气球应该慢慢充气, 直到没有进一步减少的远端地图被注意到。气球的 Overinflation 可能导致球囊破裂, 从而诱发主动脉损伤。密切注意近端和远端图有助于达到预期的主动脉球囊闭塞程度, 无论是部分或完整。
近年来, 插入鞘和主动脉球囊闭塞导管的外形变小。在这项研究中, 我们使用了14法郎插入鞘前推进的部分主动脉球囊闭塞导管 (即, 穿甲弹) 到股动脉。目前, 这一导管是在其发展的第一阶段, 计划未来的修订涉及可调气球和远端主动脉流, 以及一个较小的, 低姿态系统。然而, 较小的7法郎导管近年来已获得普及, 因为它们与较少的缺血性并发症有关。小, 低轮廓鞘和主动脉球囊闭塞导管也可以用于部署在这个失血性休克模型, 具有良好的效果。
几种出血模型用于检测大鼠失血性休克23,24,25。我们采用了一种固定容积的出血模型。在这个模型中, 一个预定的出血量, 这是基于一个计算的 TBV, 是从身体撤出在一个既定的时间段。我们使用了 35% TBV 出血超过20分钟, 这是通常用于固定容积失血性休克模型26,27,28,29。该模型广泛应用于失血性休克中休克诱发的生理变化和补偿机制, 以及病理生理反应。虽然这种方法很受欢迎, 但由于固定容积出血而引起的休克程度不同于动物。此外, 随着血体积-体重比的变化, 在这个模型中控制体重是很重要的, 以达到可重现的效果。在实践中, 其他模型类型包括固定压力出血模型、无控制出血模型和缺血性标记的出血模型作为终点。然而, 这些模型中的每一个都有自己的一套限制。
控制出血模型已用于测试主动脉球囊闭塞导管成功12。在本研究中, 我们采用了闭合性出血系统, 因为这种类型的出血模型可用于多种实验。我们的目标是为读者提供复制失血性休克模型和部署主动脉球囊闭塞导管的基础。然而, 为了创造临床上最相关和有意义的部分与完全性主动脉闭塞的比较, 这些导管最终应在设置持续远端出血的情况下进行测试。与其他创伤性侮辱, 这种模型的失血性休克可以推断出一个更临床上更现实的模型外伤损伤16,18。
动物模型外伤后的复苏策略差异很大。虽然有些人是 "流体反应性" 的拥护者--正在进行的复苏28的指导要求, 另一些则提出了管理流体 boluses 和 vasopressors2126的目标阈值。在本研究中, 我们采用阈值来确定液体丸的管理和 vasopressors 使用, 以便于重现性。虽然 "流体反应性" 复制的临床实践, 目标阈值的液体管理和 vasopressors 可能会限制广泛的变异性和主观的复苏要求的失血性休克模型。
多年来, 猪已被用于各种失血性休克模型, 提供了机会来测试广泛的治疗策略11,12,13,16,17, 19,20,21,30。然而, 重要的是要认识到, 猪不是完美的动物模型和生理变化不完全转化为人类。例如, 一些研究人员可能建议在失血性休克之前进行脾切除, 以更好地模仿人体生理学, 尽管这是有争议的话题31。
最后, 该协议证明了复制猪失血性休克模型的基本基础和主动脉球囊闭塞导管的部署。使用类似的失血性休克模型的研究结果目前正在用于调查戊酸 (VPA) 在外伤性损伤16、19、32中的作用的第二阶段临床试验,33,34. 此外, 值得注意的是, 在当今时代, 主动脉球囊闭塞导管的作用十分重要。主动脉球囊闭塞导管不仅发现了失血性休克的应用, 而且还被用于心脏和血管外科手术, 以及高危的选择性手术程序, 在这种情况下, 主动脉流的控制是有用的, 否则毁灭性的情况。总的来说, 我们认为猪失血性休克模型和主动脉球囊闭塞是高度相关的, 可以在众多的实验研究中使用。

出血仍然是在发生创伤事件的患者中可预防死亡的主要原因, 在军事环境中占90% 的创伤相关死亡, 40% 的平民人口中创伤后死亡人数为1人,2. 虽然直接压力可以治疗可压缩的出血, 但不可压缩的躯干出血仍然难以治疗, 而且如果没有迅速止血控制, 就可能致死。复苏开胸或开腹主动脉交叉夹持术的历史方法已被证明是极具侵袭性的3,4。这种干预还需要一个复杂的选择算法, 以确定的候选者, 曾遭受创伤性侮辱5。
近年来, 人们对先前所描述的方法 (REBOA)6、7、8复苏血管内气囊闭塞的兴趣再次抬头。虽然 REBOA 在出血中赋予了短期生存优势, 但在气球膨胀期间长时间完全阻断主动脉会引起严重关切, 包括不可逆转的端器官缺血9、10。为了克服这一潜在的发病率, 正在制定替代的血管内策略来管理出血。一个这样的战略, 已经看到越来越多的关注是部分闭塞的主动脉11,12。部分主动脉球囊闭塞的想法提供了血管床远端的静脉闭塞, 改善生理近端主动脉图, 并逐步后负荷减少后, 气球通缩。这些参数的变化是需要修改的生理特性的流血动物。在这种方法对人类的翻译之前, 完全和部分主动脉球囊闭塞导管已被大量测试的猪模型失血性休克11,12,13。
猪已被用于研究失血性休克多年。目前对失血性休克病理生理学的理解大多是利用动物模型 (包括猪) 的研究得出的。他们的生理和平衡反应在设置的病理体积衰竭后出血, 特别是那些有关凝血和心血管反应, 已被充分记录和像那些人14。猪失血性休克模型也为研究失血性休克和其他外伤的治疗策略提供了机会。
在本研究中, 我们展示了一个临床现实模型的猪失血性休克评估血管内治疗策略, 包括完全和部分主动脉球囊闭塞。我们推测, 部分闭塞的主动脉导致一个更好的生理和实验室的情况相比, 完全闭塞的主动脉的猪接受控制的固定容积出血。
我们的目的是比较部分和完全性主动脉闭塞作为一种治疗失血性休克猪模型的生理效应。在外伤 (穿甲弹) 导管中采用选择性主动脉球囊闭塞术 (图 1) 实现部分主动脉闭塞。穿甲弹导管是一种双气囊系统, 允许腔内血流, 从而提供了部分主动脉流到闭塞的血管床远端。完全主动脉闭塞是通过单气囊主动脉闭塞导管 (例如, 尾端) (图 1) 实现的。治疗组随机接受复苏主动脉闭塞, 无论是完全或与部分主动脉球囊闭塞导管 (n = 2/组)。
该模型的主要步骤包括诱导麻醉和插管, 维护麻醉, 仪器仪表, 35% TBV 出血 (20 分钟总; 一半以上的前7分钟, 一半以上其余13分钟), 主动脉球囊闭塞和全血复苏 (60 分钟闭塞; 20% 全血复苏在最后20分钟的闭塞), 危重监护监测 (240 分钟) 与血流动力学观察, 安乐死与组织采伐。图 2演示了本实验中使用的模型。

<table><tbody><tr><td>Yorkshire-Landrace Swine</td><td>Michigan State University Veterinary Farm</td><td></td><td></td></tr><tr><td>Anesthesia: Telazol</td><td>Pfizer</td><td></td><td>Dose: 2-8 mg/kg; IM</td></tr><tr><td>Anti-cholinergic: Atropine</td><td>Pfizer</td><td></td><td>Dose: 1mg, IM</td></tr><tr><td>Anesthesia: Isoflurane</td><td>Baxter</td><td></td><td>Dose: 1-5%, INH</td></tr><tr><td>Betadine</td><td>Humco</td><td></td><td></td></tr><tr><td>Alcohol 70%</td><td>Humco</td><td>NDC 0395-4202-28</td><td></td></tr><tr><td>Datex-Aespire Anesthesia Machine</td><td>GE Healthcare</td><td>7900</td><td></td></tr><tr><td>Endotracheal tube</td><td>DEE Veterinary</td><td>20170518</td><td>Appropriate size for animal (6.5 or 7.0F)</td></tr><tr><td>Laryngoscope</td><td>Miller</td><td>85-0045</td><td></td></tr><tr><td>Stylet</td><td>Hudson RCI</td><td>5-151--1</td><td></td></tr><tr><td>Jelco 20G IV Catheter</td><td>Smiths Medical</td><td>4054</td><td></td></tr><tr><td>Operating Room Monitor (Vital Signs Monitor)</td><td>SurgiVet Advisor</td><td>V9201</td><td>May require at least 2</td></tr><tr><td>Surgical Gowns</td><td>Kimberly Clark</td><td>90142</td><td>Use appropriate size for surgeon.</td></tr><tr><td>Sterile surgical gloves</td><td>Cardinal Health (Allegiance)</td><td>22537-570</td><td>Use appropriate size for surgeon.</td></tr><tr><td>Cautery Pencil</td><td>Medline</td><td>ESPB 2000</td><td></td></tr><tr><td>Suction tubing</td><td>Medline</td><td>DYND50251</td><td></td></tr><tr><td>Sunction tip: Yankauer</td><td>Medline</td><td>DYND50130</td><td></td></tr><tr><td>Bovie Aaron 1250 Electrocautery Unit</td><td>Bovie Medical Co. FL</td><td>BOV-A1250U</td><td></td></tr><tr><td>Salpel Blade - Size #10</td><td>Cardinal Health (Allegiance)</td><td>32295-010</td><td></td></tr><tr><td>Scalpel Handle</td><td>Martin</td><td>10-295-11</td><td></td></tr><tr><td>Debakey Forceps</td><td>Roboz</td><td>RS-7562</td><td></td></tr><tr><td>Weitlander Retractor</td><td>Roboz</td><td>RS-8612</td><td></td></tr><tr><td>Mayo Scissors</td><td>Roboz</td><td>RS-76870SC</td><td></td></tr><tr><td>Army-navy Retractor</td><td>Teleflex</td><td>164715</td><td></td></tr><tr><td>Mixter Right-angle Forceps</td><td>Teleflex</td><td>175073</td><td></td></tr><tr><td>5F (1.7 mm) 11 cm Insertion Sheath with 0.35&quot; Guidewire</td><td>Boston Scientific</td><td>16035-05B</td><td></td></tr><tr><td>8F (2.7 mm) 11 cm Insertion Sheath with 0.35&#039;&#039; Guidewire</td><td>Boston Scientific</td><td>16035-08B</td><td></td></tr><tr><td>20G angled Introducer Needle</td><td>Arrow</td><td>AK-09903-S</td><td></td></tr><tr><td>14F (4.78 mm) 13 cm Insertion Sheath with 10F dilator</td><td>Cook Medical</td><td>G08024</td><td></td></tr><tr><td>2-0 Silk 18&#039;&#039; 45 cm</td><td>Ethicon</td><td>A185H</td><td></td></tr><tr><td>3-0 Vicryl 36&#039;&#039; 90 cm</td><td>Ethicon</td><td>J344H</td><td></td></tr><tr><td>3-0 Nylon 18&#039;&#039; 45 cm</td><td>Ethicon</td><td>663G</td><td></td></tr><tr><td>4-0 Prolene 30&#039;&#039; 75 cm</td><td>Ethicon</td><td>8831H</td><td></td></tr><tr><td>20 ml syringe</td><td>Metronic/Covidien</td><td>8881512878</td><td></td></tr><tr><td>3 mL syringe</td><td>Metronic/Covidien</td><td>1180300555</td><td></td></tr><tr><td>6 mL syringe</td><td>Metronic/Covidien</td><td>1180600777</td><td></td></tr><tr><td>1000ml 0.9% Saline</td><td>Baxter</td><td>2B1324X</td><td></td></tr><tr><td>Foley Catheter (18F 30 cc)</td><td>Bard</td><td>0166V18S</td><td></td></tr><tr><td>Urinary Drainage Bag</td><td>Bard</td><td>154002</td><td></td></tr><tr><td>9F 10 cm Insertion Sheath</td><td>Arrow</td><td>AK-09903-S</td><td></td></tr><tr><td>Swan-Ganz pulmonary artery catheter (8F)</td><td>Edwards Lifesciences co. CA</td><td>746F8</td><td></td></tr><tr><td>Carotid Flow Probe System</td><td>Transonic, Ithaca, NY</td><td></td><td>3, 4, or 6 mm probes</td></tr><tr><td>SABOT catheter</td><td>Hayes Inc.</td><td></td><td></td></tr><tr><td>CODA balloon catheter</td><td>Cook Medical</td><td>8379144</td><td></td></tr><tr><td>Ultrasound, M-Turbo</td><td>SonoSite</td><td></td><td></td></tr><tr><td>Amplatz Stiff Guidewire (0.035 inch, 260 cm)</td><td>Cook Medical</td><td>G03460</td><td></td></tr><tr><td>Arterial Blood Gas Syringes</td><td>Smiths Medical</td><td>4041-2</td><td></td></tr><tr><td>Arterial Blood Gas Analyzer</td><td>Nova Biochemical</td><td>ABL800</td><td></td></tr><tr><td>Masterflex Pump</td><td>Cole Palmer</td><td>HV-77921-75</td><td></td></tr><tr><td>Blood Collection Bags</td><td>Terumo</td><td>1BBD606A</td><td></td></tr><tr><td>Macro IV drip set</td><td>Hospira</td><td>12672-28</td><td></td></tr><tr><td>Pentobarbital</td><td>Pfizer</td><td></td><td>Dose: 100 mg/kg; IV</td></tr><tr><td>Eppendorf Tubes</td><td>Sorenson</td><td>11590</td><td></td></tr><tr><td>50 cc conical tubes</td><td>Falcon</td><td>352097</td><td></td></tr><tr><td>Formalin</td><td>Fisherbrand</td><td>431121</td><td></td></tr><tr><td>Bair Hugger Normothermia System</td><td>Arizant Healthcare, Inc.</td><td></td><td></td></tr></tbody></table>

complete and partial aortic occlusion for the treatment of hemorrhagic shock in swine

出血仍然是造成创伤的可预防死亡的主要原因。非可压缩躯干出血的血管内处理是近年来创伤护理的前沿问题。由于完全性主动脉闭塞引起严重关注, 部分主动脉闭塞的概念得到了越来越多的关注。在这里, 我们提出了一个大的失血性休克动物模型, 以研究一种新的部分主动脉球囊闭塞导管的影响, 并将其与导管进行比较, 适用于完全主动脉闭塞的原则。对猪进行麻醉和检测, 以进行控制的固定容积出血, 并监测血流动力学和生理参数。出血后, 主动脉球囊闭塞导管插入和充气在 supraceliac 主动脉60分钟, 在此期间, 动物接受全血复苏为20% 的总血容量 (TBV)。在气球通缩后, 动物被监测在一个关键的护理设置4小时, 在此期间, 他们接受液体复苏和 vasopressors 的需要。部分主动脉球囊闭塞显示在气球充气期间改善了远端平均动脉压力 (map), 减少了缺血标志物, 减少了液体复苏和升压使用。猪失血性休克模型可以用来测试各种治疗策略, 因为出血后的猪生理和稳态反应已经有了很好的记录, 和人类一样。除了治疗出血, 主动脉球囊闭塞导管已成为流行的作用, 在心脏骤停, 心脏和血管外科手术, 以及其他高危的选择性手术程序。

血流动力学和生理参数:
该地图在出血后立即减少 (图 3A - 3D)。在气球充气阶段, 完全闭塞组的动物比部分闭塞组的动物经历了更高的近端图 (图 3A和3B)。与完全闭塞组 (平均远端图, 气球充气阶段相比, 局部闭塞组的平均远端图为高; 部分:31 2.9 mmHg, 完整: 16.5 @ 1.14 mmHg;p > 0.05), 反映部分远端主动脉流 (图 3C和3D)。复苏后, 近端和远端图在两组增加, 并返回到基线后, 气球通货紧缩的其余的关键照顾阶段 (图 3A - 3D)。
所有动物都在出血后立即发生反射性心动过速, 这在两组气球充气阶段都有递增的增长 (图 4A)。在球囊通缩后, 全闭塞组的危重监护期其余部分的时间明显高于部分闭塞组, 虽然 HR 的这种差异没有统计学意义。
出血后, CVP 减少两组 (图 4B)。在气球膨胀之后, 它经历了一个上升的趋势。随着气球的通缩, 完全闭塞组显示, CVP 比部分闭塞组更大的减少, 虽然没有统计学上的差异。在关键护理阶段的额外复苏后, CVP 恢复到两组的基线。同样地, CO 在出血以后减少了, 在气球通货膨胀期间增加了, 并且回到了基线在气球通缩和复苏以后为两个小组 (图 4C)。
出血后, 两组颈动脉血流均减少 (图 4D)。随着气球的膨胀, 完全闭塞组表现出较高的颈动脉血流率与部分闭塞组。随着复苏和气球收缩, 颈动脉流速恢复到基线的两组。然而, 与部分闭塞组相比, 全闭塞组颈动脉血流低。
实验室参数:
各组在基线 pH 值和乳酸水平上均无明显差异。在气球膨胀之后, 两组动物的 pH 值都有下降 (图 5A)。完全闭塞组的 pH 值明显低于部分闭塞组 (完整: 7.14 0.01, 部分: 7.32 @ 0.02, p = 0.1)。乳酸水平明显高于整个气球的通货膨胀和关键照顾阶段的其余部分完全闭塞组 (完整: 17.5 0.71 毫摩尔, 部分: 6.1 @ 0.28 毫摩尔, p = 0.03) (图 5B)。这种乳酸水平的差异缓慢下降, 直到在关键护理阶段结束时的水平相似。
复苏要求:
完全闭塞组动物的总体液需求量明显高于部分闭塞组动物 (完全闭塞组动物的总补充液体复苏: 47.5 3.4 厘米3/千克,部分闭塞组动物的补充液体复苏总数: 3.7, 0.4 厘米3/千克, p = 0.003) (图 6A)。同样, 完全闭塞组的去甲肾上腺素要求明显高于部分闭塞组 (完整: 289.7 25.4 µg/千克, 部分:32 @ 13.8 µg/千克, p = 0.006) (图 6B)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58284/58284fig1.jpg"> 图 1: 主动脉球囊闭塞导管.(A) 部分主动脉闭塞是利用外伤 (穿甲弹) 导管中选择性主动脉球囊闭塞而实现的, 而完全主动脉闭塞是使用完全主动脉球囊闭塞导管实现的。(B) 部分主动脉球囊闭塞导管是一种双气囊系统, 允许腔内血流提供远端主动脉流。完整的主动脉闭塞使用单气囊系统提供。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/58284/58284fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58284/58284fig2.jpg"> 图 2: 伤害协议.一项由35% 血总容积出血引起的损伤, 其次为1小时的主动脉球囊闭塞。在40分钟的气球闭塞后, 20% 全血20分钟进行复苏。在气球通缩后的4小时内, 这些动物被监控在关键的护理阶段。基本法= 基线;PS = 后冲击;PR = 复苏后的时期。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/58284/58284fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58284/58284fig3.jpg"> 图 3: 对损伤和气球膨胀的血流动力学反应.这些面板显示了 (A) 近端 pean 动脉压 (图) 的术中测量, (B) 气球充气过程中的近端图, (C) 远端图, (D) 气球中的远端图通胀。数据显示为组的平均值, 即标准误差 (SE)。S = 冲击期间 (20 分钟);气球= 气球通货膨胀 (60 分钟);R = 复苏 (20 分钟);PR = 复苏后时期/气球通缩;E = 受伤阶段的结束 (在休克期完成后5小时);完成= 完全主动脉球囊闭塞导管;部分= 部分主动脉球囊闭塞导管。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/58284/58284fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58284/58284fig4.jpg"> 图 4: 对受伤和气球部署的系统性和生理反应.这些面板显示的术中测量 (A) 心率 (HR), (B) 中心静脉压力 (CVP), (C) 心脏输出 (CO) 和 (D) 颈动脉血流 (CF)。数据显示为组均值S = 休克期 (20 分钟);气球= 气球通货膨胀 (60 分钟);R = 复苏 (20 分钟);PR = 复苏后时期/气球通缩;E = 受伤阶段的结束 (在休克期完成后5小时);完成= 完全主动脉球囊闭塞导管;部分= 部分主动脉球囊闭塞导管。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/58284/58284fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/58284/58284fig5.jpg"> 图 5: 针对损伤和气球充气的实验室参数.这些面板显示了 (A) pH 值和 (B) 乳酸的术中测量.数据显示为组平均值。星号表示有显著不同的时间点 (p < 0.05)。S = 冲击期间 (20 分钟);气球= 气球通货膨胀 (60 分钟);R = 复苏 (20 分钟);PR = 复苏后时期/气球通缩;E = 损伤阶段的结束 (在休克期完成后5小时)。完成= 完全主动脉球囊闭塞导管;部分= 部分主动脉球囊闭塞导管。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/58284/58284fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/58284/58284fig6.jpg"> 图 6: 针对受伤和气球充气的复苏要求.这些面板显示的术中测量 (A) 总额外的复苏液和 (B) 去甲肾上腺素的使用.数据显示为组平均值。星号表示显著差异 (p < 0.05)。完成= 完全主动脉球囊闭塞导管;部分= 部分主动脉球囊闭塞导管。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/58284/58284fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Complete and Partial Aortic Occlusion for the Treatment of Hemorrhagic Shock in Swine at JoVE.com

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

在这里, 我们提出了一个协议, 以证明一个失血性休克模型的猪, 使用主动脉闭塞作为桥梁, 最终护理的创伤。该模型在测试多种外科和药理治疗策略中有应用。

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 ...

complete-partial-aortic-occlusion-for-treatment-hemorrhagic-shock

Universidad San Sebastian

Research

JoVE Journal

Medicine

10.5K 次观看.  University of Michigan. 在这里, 我们提出了一个协议, 以证明一个失血性休克模型的猪, 使用主动脉闭塞作为桥梁, 最终护理的创伤。该模型在测试多种外科和药理治疗策略中有应用。

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

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

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

原位主动脉瓣移植在小鼠血管疾病的研究

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

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 ...

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 ...

Development of a Selective Aortic Arch Perfusion System in a Porcine Model of Exsanguination Cardiac Arrest

Hemorrhage constitutes the majority of potentially preventable deaths from trauma. There is growing interest in endovascular resuscitation techniques such as selective aortic arch perfusion (SAAP) for patients in cardiac arrest. This involves active perfusion of the coronary circulation via a thoracic aortic balloon catheter and is approaching clinical application. However, the technique is complex and requires refinement in animal models before human use can be considered. This paper describes a large animal model of exsanguination cardiac arrest treated with a bespoke SAAP system.
Swine were anesthetized, instrumented and a splenectomy was performed before a controlled, logarithmic exsanguination was initiated. Animals were heparinized and the shed blood collected in a reservoir. Once cardiac arrest was observed, the blood was pumped through an extra-corporeal circuit into an oxygenator and then delivered through a 10 Fr balloon catheter placed in the thoracic aorta.
This resulted in the return of a spontaneous circulation (ROSC) as demonstrated by ECG and aortic root pressure waveform. This model and accompanying SAAP system allow for standardized and reproducible recovery from exsanguination cardiac arrest.

<meta charset="utf8"></head><body>在猪出血性心脏骤停模型中开发选择性主动脉弓灌注系统

The goal of this protocol is to demonstrate a porcine exsanguination cardiac arrest model and a specifically built selective aortic arch perfusion circuit for translational research.

在猪出血性心脏骤停模型中开发选择性主动脉弓灌注系统

Hemorrhage constitutes the majority of potentially preventable deaths from trauma. There is growing interest in endovascular resuscitation techniques such ...

Post-Myocardial Infarction Heart Failure in Closed-chest Coronary Occlusion/Reperfusion Model in G&#246;ttingen Minipigs and Landrace Pigs

The development of heart failure is the most powerful predictor of long-term mortality in patients surviving acute myocardial infarction (MI). There is an unmet clinical need for prevention and therapy of post-myocardial infarction heart failure (post-MI HF). Clinically relevant pig models of post-MI HF are prerequisites for final proof-of-concept studies before entering into clinical trials in drug and medical device development.
Here we aimed to characterize a closed-chest porcine model of post-MI HF in adult G&#246;ttingen minipigs with long-term follow-up including serial cardiac magnetic resonance imaging (CMRI) and to compare it with the commonly used Landrace pig model.
MI was induced by intraluminal balloon occlusion of the left anterior descending coronary artery for 120 min in G&#246;ttingen minipigs and for 90 min in Landrace pigs, followed by reperfusion. CMRI was performed to assess cardiac morphology and function at baseline in both breeds and at 3 and 6 months in G&#246;ttingen minipigs and at 2 months in Landrace pigs, respectively.
Scar sizes were comparable in the two breeds, but MI resulted in a significant decrease of left ventricular ejection fraction (LVEF) only in G&#246;ttingen minipigs, while Landrace pigs did not show a reduction of LVEF. Right ventricular (RV) ejection fraction increased in both breeds despite the negligible RV scar sizes. In contrast to the significant increase of left ventricular end-diastolic (LVED) mass in Landrace pigs at 2 months, G&#246;ttingen minipigs showed a slight increase in LVED mass only at 6 months.
In summary, this is the first characterization of post-MI HF in G&#246;ttingen minipigs in comparison to Landrace pigs, showing that the G&#246;ttingen minipig model reflects post-MI HF parameters comparable to the human pathology. We conclude that the G&#246;ttingen minipig model is superior to the Landrace pig model to study the development of post-MI HF.

The overall goal of the current study is to present the techniques of induction of myocardial infarction (MI) and post-myocardial infarction heart failure (post-MI HF) in closed-chest, adult G&#246;ttingen minipigs and the characterization of post-MI HF model in G&#246;ttingen minipigs as compared to Landrace pigs.

封闭胸冠状动脉闭塞/输液模型中的心肌梗塞后心力衰竭

The development of heart failure is the most powerful predictor of long-term mortality in patients surviving acute myocardial infarction (MI). There is an ...

Myocardial Infarction by Percutaneous Embolization Coil Deployment in a Swine Model

Myocardial infarction (MI) is the leading cause of mortality worldwide. Despite the use of evidence-based treatments, including coronary revascularization and cardiovascular drugs, a significant proportion of patients develop pathological left-ventricular remodeling and progressive heart failure following MI. Therefore, new therapeutic options, such as cellular and gene therapies, among others, have been developed to repair and regenerate injured myocardium. In this context, animal models of MI are crucial in exploring the safety and efficacy of these experimental therapies before clinical translation. Large animal models such as swine are preferred over smaller ones due to the high similarity of swine and human hearts in terms of coronary artery anatomy, cardiac kinetics, and the post-MI healing process. Here, we aimed to describe an MI model in pig by permanent coil deployment. Briefly, it comprises a percutaneous selective coronary artery cannulation through retrograde femoral access. Following coronary angiography, the coil is deployed at the target branch under fluoroscopic guidance. Finally, complete occlusion is confirmed by repeated coronary angiography. This approach is feasible, highly reproducible, and emulates the pathogenesis of human non-revascularized MI, avoiding the traditional open-chest surgery and the subsequent postoperative inflammation. Depending on the time of follow-up, the technique is suitable for acute, sub-acute, or chronic MI models.

Myocardial infarction (MI) animal models that emulate the natural process of the disease in humans are crucial to understanding pathophysiological mechanisms and testing the safety and efficacy of new emergent therapies. Here, we describe an MI swine model created by deploying a percutaneous embolization coil.

猪模型中经皮栓塞线圈部署的心肌梗死

Myocardial infarction (MI) is the leading cause of mortality worldwide. Despite the use of evidence-based treatments, including coronary revascularization ...

Complete and Partial Resuscitative Endovascular Balloon Occlusion of the Aorta for Hemorrhagic Shock

Resuscitative endovascular balloon occlusion of the aorta (REBOA) devices grew out of a military-civilian partnership to develop new capabilities for hemorrhage control. With the advent of purpose-built devices, REBOA has become increasingly common in civilian trauma and acute care settings. Currently available REBOA catheters were designed as complete aortic occlusion devices. However, the therapeutic window for complete aortic occlusion is time-limited due to ischemia-reperfusion injury. The partial procedure allows blood flow past the level of occlusion while maintaining targeted proximal pressure, which has been shown to reduce distal ischemia and adjunctive resuscitation requirements in preclinical studies with prolonged occlusion times as compared to traditional complete occlusion.
pREBOA-PRO is the first catheter designed to enable partial and complete aortic occlusion and is currently in limited market release at seven Level I trauma centers in North America. This paper will focus on procedural considerations for REBOA, including patient selection criteria and a comparison of complete and partial aortic occlusion in a simulator, along with highlighting critical steps to improve clinical outcomes. Additionally, this paper reviews a contrast-enhanced CT scan from a trauma patient that shows distal perfusion after 2 h of partial aortic occlusion using this newly designed catheter and discusses representative results from the limited market release to highlight the profound effect of technological innovation on outcomes in vascular emergencies.

A commercial catheter was designed to facilitate true partial resuscitative endovascular balloon occlusion of the aorta (REBOA) and address the complications associated with complete aortic occlusion. Initial clinical reports indicate that partial REBOA improves transition to reperfusion, reduction of distal ischemia, and extension of safe occlusion time compared to complete occlusion.

完全和部分复苏性主动脉血管内球囊闭塞治疗出血性休克

Resuscitative endovascular balloon occlusion of the aorta (REBOA) devices grew out of a military-civilian partnership to develop new capabilities for ...

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 ...

完全性和部分主动脉闭塞治疗猪失血性休克

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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

原位主动脉瓣移植在小鼠血管疾病的研究

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

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

在猪出血性心脏骤停模型中开发选择性主动脉弓灌注系统

在猪出血性心脏骤停模型中开发选择性主动脉弓灌注系统

封闭胸冠状动脉闭塞/输液模型中的心肌梗塞后心力衰竭

猪模型中经皮栓塞线圈部署的心肌梗死

完全和部分复苏性主动脉血管内球囊闭塞治疗出血性休克

容量控制性失血性休克的猪婴儿模型的描述