按照国家机构的研究和所有动物协议的俄勒冈卫生科学大学的体制的动物护理和使用委员会批准的护理和使用动物的健康指引所述的所有程序进行。 1。手术准备<ol><li>称量鼠标。描述的过程进行C57BL / 6小鼠体重20至25克使用3-4％的异氟醚诱导麻醉诱导，并随后保持1.5-2.5％异氟醚在空气/氧气混合。 </li><li>润滑眼睛的位置上加热垫的动物仰卧。用胶布固定4四肢。后腿的爪子可能是在一个中立的位置录音，但是，前爪应该得到保护尽可能靠近胸壁允许在胸外按压胸壁游览。 </li><li>润滑和地方直肠温度探头。温度控制采用加热垫和灯连接到电子温度控制器（从DIGI -感，科尔帕默，弗农山白细胞介素），这是保持37.0 ° C。因为它可能是动物内的温度梯度发展在无流量状态，重要的是，温度进行测量和控制利益器官附近的。 </li><li>使用一个2.5厘米22 GA聚四氟乙烯导管（Insyte W，屋宇署，新泽西州富兰克林）和切割的角度介绍人（Frova介绍人，库克医疗，布卢明顿IN）的末端插管气管。气管插管的其他方法是可以接受的的，但是，使用的角度介绍人允许在一个位置略有延长，优化放置静脉留置针的手术暴露颈部的正确定位。气管导管枢纽的安全与缝合门齿的循环，并保持轻微的张力固定在胸外按压头部。 </li><li>机械通风与鼠标设置为140μL，150次/分钟的啮齿类动物呼吸。这是不调整的体重总是在20-25克动物作为本议定书的执行。不要过度加深麻醉。不要设置呼吸机申请结束，因为这损害复苏呼气压力。 </li><li>使用无菌技术和手术显微镜，前刷新PE - 10导管在颈内静脉。一方可能被使用，但使用最接近经营者的一侧，减少导管移位在胸外按压或其他操作的机会。 0.5％布比卡因，0.1毫升到伤口边缘是infitrated控制术后疼痛。 </li><li> PE - 10导管固定到皮肤缝合与手术氰基丙烯酸酯胶粘剂。 </li><li>放置皮下心电图电极，并连接到监控设备。仔细留意的信号路径和信号本身的最大化，是复苏成功的关键。确保所有电线固定作业面，尽量减少信号口岸，并尽量减少信号路径内的绝缘体（如空气空心针或绞线）。 nonstranded导线连接的固体针可购买或在实验室中制成。一旦连接，优化心电图信号在监视器上。 </li></ol> 2。心脏骤停<ol><li>确保鼠标是常温下的定义，直肠温度36.5 ° C - 37.5 ° C。管理40μL室温0.5 M的氯化钾静脉注射和心电图跟踪观察等电。体重调整剂量是没有必要的范围内20-25克。开始逮捕定时器。 </li><li>断开呼吸机。停止麻醉蒸汽。关闭电热垫和任何其他设备产生的电子噪音，可能会干扰心电图监测。鼠标放在一个绝缘毯。 </li><li>记录温度每分钟心脏骤停期间。如果有必要，加热灯可用​​于把核心温度常温范围。在心脏骤停，准备用品和设备（例如，肾上腺素注射器）复苏。可以帮助一个清单，以确保不间断的复苏，这是生存所必需的。 </li><li> 7分钟后，心脏骤停30秒，重新呼吸机和180次/分，增加的速度保持在逮捕前设置的潮气量。 </li><li>在第8分钟，300 BPM开始胸外按压。运动工件上的心电图可以用来判断心肺复苏率。胸外按压应交付与食指，剑突以上的5毫米，略向中线的左侧。胸部要压缩的前后距离的1/3-1/2，必须充分反冲按压之间允许。手指的定位和最佳的压缩压力绝对是至关重要的。未能达到生存在这个模型几乎总是由于不理想的心肺复苏。 </li><li>注入0.5毫升的肾上腺素，稀释至15微克/毫升，在心肺复苏的前30秒，。共有肾上腺素剂量为8-12微克。仔细观察自主循环（ROSC）的回报率的心电图。复杂窄QRS波群之间的压缩失真。 ROSC的，通常会出现介于90秒和2分钟后开始心肺复苏。复苏是被遗弃，如果ROSC的不发生3分钟。频发室性早搏和心电图轴的变化，观察在ROSC后第2分钟，几乎总是在2分钟内解决稳定的窦性心动过速。 </li><li>记录的复苏和肾上腺素剂量的总时间。记录温度，每分钟10分钟后，ROSC的。 </li><li>心电图信息可以去掉开始自主呼吸时，通常在12-15分钟后ROSC的。 </li><li>拔管气管时，自发呼吸频率&gt; 60/min。 </li><li>恢复笼放置在鼠标上的一个温度控制的表面设置为37 ° C的首2小时postprocedure，确保容易获得食物和水。笼子，然后被转移到标准的术后住房条件，当鼠标从麻醉中完全恢复，是活跃。 </li></ol> 3。放血/灌注固定和肾脏丰收<ol><li> 24小时后的CA / CPR，诱导麻醉与异氟醚的3-4％，和安全的动物，在仰卧位手术表面上在通风柜这是适合用福尔马林.. </li><li>深化麻醉剂麻醉蒸气的浓度逐渐增加至5％，并确保鼠标深受麻醉，停止自主呼吸。 </li><li>执行翻盖开胸，并按照标准技术的心脏左心室心尖穿刺抽血通过鼠标。 </li><li>通过同一个针头，管理0.9％生理盐水缓慢静滴。 acclerate灌注/放血，尼克用剪刀右心耳。执行剖腹探查术，并观察肾脏。当他们被吓得脸色煞白，生理盐水，可能会更改为4％的福尔马林对固定。肾脏，然后嵌入在石蜡和氟乙玉肾小管上皮细胞坏死的定量染色。 </li></ol> 4。代表性的成果: 当CA诱导，平均动脉压（MAP）和区域肾血流量（RRBCF）下降到接近零，并保持稳定，直到复苏开始（ 图 1 ）。 24小时后CA /心肺复苏术，肾功能（尿素氮，尿素氮和肌酐）血清指数明显升高，与假手术动物。 AST / ALT升高，深刻全身缺血（ 图2）提供的证据。嗜中性粒细胞明胶酶相关载脂蛋白（NGAL），肾缺血性损伤的敏感指标是大规模上调24小时后的CA / CPR（ 图3） 。最后，显微照片表明典型的肾小管上皮变薄和苏木精和曙红管腔充盈染色（ 图4中，A组 ）和广泛髓质肾小管上皮细胞染色与玉氟乙死亡时的片状，髓坏死，缺血性肾损伤（ 图4，B组 ）。 <img src="/files/ftp_upload/3116/3116fig1.jpg" alt="图1" /> 图1。心脏骤停引起的灌注压的瞬间失去，这里代表平均动脉压（MAP），在股动脉测量，造成接近完成整个心脏骤停期间停止区域肾皮质血流量（RRCBF） （阴影区）。复苏胸外按压和肾上腺素返回映射到正常RRCBF稳步上升后的复苏期。 来自14个许可转载 。 <img src="/files/ftp_upload/3116/3116fig2.jpg" alt="图2" /> 图2。postprocedure 24小时，血中尿素氮（BUN），肌酐，肾小管上皮细胞死亡的程度都显着升高的CA / CPR与假手术过程中处理的动物相比，动物经过。 CA / CPR诱导泛有机体的缺血性损伤，肝功能酶的大规模提升证明丙氨酸氨基转移酶（ALT）和谷草转氨酶（AST），CA / CPR小鼠与假治疗的动物相比。 <img src="/files/ftp_upload/3116/3116fig3.jpg" alt="图3" /> 图3 Western印迹进行多克隆抗体，以中性粒细胞明胶酶相关载脂蛋白（NGAL），肾缺血性损伤的敏感指标。尿液样本立即获得前（"前"）和24小时后（"24小时"）4只动物的CA / CPR（标记为A，B，C和D以上）。 NGAL的大规模上调鼠标CA / CPR后的尿液。转载来自14个许可。 <img src="/files/ftp_upload/3116/3116fig4.jpg" alt="图4" /> 图4 A）苏木精和曙红染色的CA / CPR后第一个肾组织24小时的肝门部短轴。有髓质和髓质肾小管，肾小管堵塞的修修补补，但明确的损害。箭头指向与肿，细胞核固缩在皮髓质交界处损坏肾小管。二）Flouro玉乙染色同一地区在同一动物的CA / CPR后24小时。玉氟乙污渍的坏死细胞明亮的绿色，呈片状皮髓质肾小管坏死。箭头指向明亮的染色在皮髓质交界处损坏管。这些结果是从人类发展AKI的肾活检结果大致相似，而不像AKI的其他动物模型制作的。

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没有利益冲突的声明。

常温下在小鼠心脏骤停和心肺复苏模型提供了一个模式，重复的病理生理及临床最常见的急性肾损伤，全身血流灌注不足引起的形态的多种渠道的评价。假设检验，可访问的遗传操作技术和充分理解和特点的解剖和生理学实验小鼠的一整套资助。 这里描述的，对于有经验的生存率是80％。虽然手术的准备和逮捕后的护理很简单，被捕鼠标的实际复苏是一个具有挑战性的手术技能，需要大量的实践。根据我们的经验，成功resuscitations成为常态后约一个没有经验的外科医生在急救动物个体的第一个30-50尝试，和一致的损伤和复苏采取其他做法。 ，特别是在学习的技术（即第20个动物）的早期阶段，有意义的生存是不可能的，任何生还者，应立即复苏后安乐死，以避免不必要的困扰。早期训练后，幸存者展示活力的复苏分钟的心脏复苏时，它是适当的继续幸存的动物通过15分钟后ROSC的标志。证明这一点大力自发呼吸努力的动物可能会适当的恢复与生存24小时。 IACUC协议，这种做法阶段应予以具体阐述。最困难的技能学习是胸外按压本身，因为它是难以提供所需的速度，没有有害的水平越来越大的压力按压。机械复苏已经在我们的实验室和其他 16，但制订和使用，迄今为止还没有任何机械的人工呼吸器，鼠标生产可以接受的生存。 这种模式有一些限制。首先，没有小鼠模型可以完全模拟人类生理，结果必须得到尊重解释为物种间的差异。尤其是鼠标的心脏是强大的，我们实现80％的生存率在心脏骤停8分钟，但在临床研究中，人类的生存是低于50％，甚至逮捕后开始复苏 2-3分钟17鼠标体积小。呈现在技术上严格的程序，并有一个显着的学习曲线，特别是胸外按压，的。其次，在本议定书中，药物剂量和呼吸机的设置没有索引的动物体重。这是因为我们使用20-25克重范围只动物，以尽量减少设备的大小的影响。例如，我们用一个22号导管，气管插管;取得的印章是在30克的鼠标不相同，因为它是在25克鼠标，增加煤气泄漏可能是生理重大。但是，动物在规定重量范围内使用，不如实地再现各种临床缺血再灌注损伤的身体类。第三，无流动状态未必能够忠实再现低流量状态的结果。特别是，温度梯度发展内鼠标无流量期间。为了最大限度地减少梯度对结果的影响（如有），温度测量附近器官的利益。最后，由于我们的实验室证实postarrest期间显着的代谢性酸中毒的早期数据，我们哽咽着在复苏。由于最近的数据表明，过度换气，在心肺复苏术是有害的，18,19是可能的，这种做法减少的生存模式。在许多案例中，全身缺血再灌注损伤模型的替代品存在。例如，CA / CPR已被描述的狗，20只兔子，21羔羊，22头猪，23只大鼠，16日和北极地松鼠24 。 我们已经表明，术后肾功能可能是由一些化学，免疫学，组织学技术，或在24小时内评估。动物可以存活时间超过24h，然而，AKI的瞬态性质使得这种模式有吸引力的一个用于恢复和/或永久性的损伤机制的调查。伤害本身是通过改变在CA的时间和温度参数滴定。更长的逮捕和维持较高的温度会产生更大的伤害（但产生不利影响生​​存）。然而，正如我们已经与包括肝功能数据显示，其他器官系统也受到影响，并可能使用这种模式适应调查​​，重大的努力可能需要优化实验条件下的利益的器官系统。

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th></tr><tr><td>氯化钾</td><td>雅培医疗</td><td> 06653-5 </td></tr><tr><td>异氟醚</td><td>雅培医疗</td><td> 05260-05（和其他） </td></tr><tr><td>肾上腺素盐酸</td><td>多</td><td>多</td></tr><tr><td>从DIGI -感温度控制器</td><td>科尔 - 帕默</td><td> EW - 89000 - 00 </td></tr><tr><td> Angiocath四蛋白酶</td><td> VWR </td><td> 381134 </td></tr><tr><td> Frova角度呼吸道介绍人</td><td>厨师</td><td> G27282 </td></tr><tr><td>对小鼠的MicroVent换气扇</td><td>哈佛器械</td><td> 733591 </td></tr></tbody></table>

normothermic cardiac arrest and cardiopulmonary resuscitation: a mouse model of ischemia-reperfusion injury

5,6全身低灌注成功繁殖，在啮齿类动物模型急性肾损伤（AKI）是一个共同的，极具杀伤力，危重病的并发症， 死亡率高 1-4，这是最经常全身血流灌注不足引起的。一直困难重重。7-9,9,10聘请局灶性脑缺血模型反复论证的结果没有转化为临床上，和较大的动物模型，全身血流灌注不足无法获得的遗传操纵的工具在鼠标的可能。11,12然而，心脏骤停和心肺复苏的小鼠模型，近年来又出现了可适应模型 AKI的13这种模式可靠地再现生理功能，解剖和组织学在临床AKI的看到的结果，迅速重复，并提供所有手术小鼠模型的显着优点，包括访问的基因操纵技术，成本相对低的大型动物，和易用性。使用这种模式，集团已开发的丰富经验，以评估在AKI的一些器官的具体成果。 14,15

Watch this Scientific Journal Video about Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury at JoVE.com

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

一个强大的围手术期和关键护理相关的急性肾损伤模型。使用近复制临床AKI的组织学和功能的变化有可能诱发心脏骤停的全身血流灌注不足。

常温心脏骤停和心肺复苏术:缺血再灌注损伤的小鼠模型

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

normothermic-cardiac-arrest-cardiopulmonary-resuscitation-mouse-model

Nanyang Technological University

Research

JoVE Journal

Medicine

18.0K 次观看.  Oregon Health & Sciences University. 一个强大的围手术期和关键护理相关的急性肾损伤模型。使用近复制临床AKI的组织学和功能的变化有可能诱发心脏骤停的全身血流灌注不足。

视频: Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

急性肾损伤和伤后纤维化小鼠缺血再灌注模型

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

科研

医学

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for myocardial infarction and cardiac complications of thoracic surgery. The ability of certain drugs to precondition the myocardium against ischemia reperfusion injury has led to multiple clinical trials, with little success. The isolated heart model allows acute observation of the functional effects of ischemia reperfusion injury in real time, including the effects of various pharmacological interventions administered at any time-point before or within the ischemia-reperfusion injury window. Since brief periods of ischemia can precondition the heart against ischemic injury, in situ aortic cannulation is performed to allow for functional assessment of non-preconditioned myocardium. A saline filled balloon is placed into the left ventricle to allow for real-time measurement of pressure generation. Ischemic injury is simulated by the cessation of perfusion buffer flow, followed by reperfusion. The duration of both ischemia and reperfusion can be modulated to examine biochemical events at any given time-point. Although the Langendorff isolated heart model does not allow for the consideration of systemic events affecting ischemia and reperfusion, it is an excellent model for the examination of acute functional and biochemical events within the window of ischemia reperfusion injury as well as the effect of pharmacological intervention on cardiac pre- and postconditioning. The goal of this protocol is to demonstrate how to perform in situ aortic cannulation and heart excision followed by ischemia/reperfusion injury in the Langendorff model.

The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

诱导和缺血再灌注损伤评估中的Langendorff灌注鼠心

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

肠缺血再灌注损伤的小鼠模型

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, diabetic retinopathy, and retinal vascular occlusions. Rodent models of I/R injury are providing significant insights into mechanisms and treatment strategies for human I/R injury, especially with regard to neurodegenerative damage in the retinal neurovascular unit. Presented here is a protocol for inducing retinal I/R injury in mice through elevation of intraocular pressure (IOP). In this protocol, the ocular anterior chamber is cannulated with a needle, through which flows the drip of an elevated saline reservoir. Using this drip to raise IOP above systolic arterial blood pressure, a practitioner temporarily halts inner retinal blood flow (ischemia). When circulation is reinstated (reperfusion) by removal of the cannula, severe cellular damage ensues, resulting ultimately in retinal neurodegeneration. Recent studies demonstrate inflammation, vascular permeability, and capillary degeneration as additional elements of this model. Compared to alternative retinal I/R methodologies, such as retinal arterial ligation, retinal I/R injury by elevated IOP offers advantages in its&#160;anatomical specificity, experimental tractability, and technical accessibility, presenting itself as a valuable tool for examining neuronal pathogenesis and therapy in the retinal neurovascular unit.

This article describes a procedure for inducing retinal ischemia-reperfusion injury by elevated intraocular pressure in mice. Retinal ischemia-reperfusion injury by elevated intraocular pressure serves to model human pathologies characterized by compromised oxygen and nutrient delivery in the retina, enabling researchers to examine potential cellular mechanisms and treatments for human diseases of the retinal neurovascular unit.

视网膜缺血再灌注损伤的小鼠模型通过眼压升高

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses

Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent re-oxygenation of the tissues (reperfusion). In the skin, ischemia-reperfusion (IR) is the main contributing factor to the pathophysiology of pressure ulcers. While the cascade of events leading up to the inflammatory response has been well studied, the spatial and temporal responses of the different subsets of immune cells to an IR injury are not well understood. Existing models of IR using the clamping technique on the skin flank are highly invasive and unsuitable for studying immune responses to injury, while similar non-invasive magnet clamping studies in the skin flank are less-than-ideal for intravital imaging studies. In this protocol, we describe a robust model of non-invasive IR developed on mouse ear skin, where we aim to visualize in real-time the cellular response of immune cells after reperfusion via multiphoton intravital imaging (MP-IVM).

This protocol describes the induction of an ischemia-reperfusion (IR) model on mouse ear skin using magnet clamping. Using a custom-built intravital imaging model, we study in vivo inflammatory responses post-reperfusion. The rationale behind the development of this technique is to extend the understanding of how leukocytes respond to skin IR injury.

在鼠耳皮肤的免疫反应的活体多光子成像诱导缺血再灌注损伤

Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent ...

Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse model is useful for investigating the pathophysiology of cerebral ischemia-reperfusion and determining potential therapeutic strategies for patients with stroke. Variations in the anatomy of the circle of Willis of C57BL/6 mice affects their infarct volume after cerebral-ischemia-induced injury. Studies have indicated that distal MCA occlusion (MCAO) can overcome this problem and result in a stable infarct size. In this study, we establish a two-vessel occlusion mouse model of cerebral ischemia-reperfusion through the interruption of the blood flow to the right MCA. We distally ligate the right MCA and right common carotid artery (CCA) and restore blood flow after a certain period of ischemia. This ischemia-reperfusion injury induces an infarct of stable size and a behavioral deficit. Peripheral immune cells infiltrate the ischemic brain within the 24 h infiltration period. Additionally, the neuronal loss in the cortical area is less for a longer reperfusion duration. Therefore, this two-vessel occlusion model is suitable for investigating the immune response and neuronal recovery during the reperfusion period after cerebral ischemia.

A mouse model of cerebral ischemia-reperfusion is established to investigate the pathophysiology of stroke. We distally ligate the right middle cerebral artery and right common carotid artery and restore blood flow after 10 or 40 min of ischemia.

脑缺血再灌注的双血管闭塞小鼠模型

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse ...

Ultrasonographic Assessment During Cardiopulmonary Resuscitation

The US-CAB (Ultrasound, Circulation/Airway/Breathing) protocol integrates several sonographic techniques into a structured assessment of the circulation, airway, and breathing status of a patient during cardiopulmonary resuscitation (CPR) in an advanced life support-compliant manner. US-C provides a subxiphoid view of the heart, to look for potentially reversible causes of disease, such as pericardial effusion, pulmonary embolism, hypovolemia, and acute coronary thrombosis. Sonographic cardiac activity during CPR not only helps differentiate pseudo-pulseless electrical activity (PEA) from true PEA but also represents a higher chance of the return of spontaneous circulation (ROSC) and survival. Evaluation of the inferior vena cava (IVC) shows the fluid status of the patient and indicates the best methods to use for fluid resuscitation. If aortic dissection is suspected, a subxiphoid view of the aorta is suggested for identifying an intimal flap. Once intubation is done, tracheal ultrasound (US-A) at the suprasternal notch helps differentiate endotracheal intubation (one air-mucosal interface with one comet-tail) from esophageal intubation (double tract sign). Immediately following US-A, bilateral lung US (US-B) should be done to confirm proper bilateral ventilation using the lung sliding sign. In addition, US-C can be serially followed to see the dynamic changes in the cardiac chambers and IVC, or any cardiac contraction suggestive of ROSC. US-B can also detect coexisting lung or pleural pathologies without interfering with the performance of CPR. The main concern when implementing this method is maintaining high-quality CPR without delays in chest compressions when performing US-CAB. Rigorous training and continued practice are key to minimize any interruptions during resuscitation.

Here, we present a US-CAB (Ultrasound, Circulation/Airway/Breathing) protocol for use during cardiopulmonary resuscitation (CPR). US-C evaluates the subxiphoid view of the heart and inferior vena cava. After intubation, tracheal US (US-A) and lung US (US-B) help confirm endotracheal intubation and proper ventilation.

心肺复苏期间的超分谱评估

The US-CAB (Ultrasound, Circulation/Airway/Breathing) protocol integrates several sonographic techniques into a structured assessment of the circulation, ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

重建显微外科缺血再灌注损伤研究的临床前大鼠模型

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, cardiac stem cell therapy is studied by delivering SCs concurrently with the induction of myocardial injury. However, this approach presents two significant limitations: the early hostile pro-inflammatory ischemic environment may affect the survival of transplanted SCs, and it does not represent the subacute infarction scenario where SCs will likely be used. Here we describe a two-part series of surgical procedures for the induction of ischemia-reperfusion injury and delivery of mesenchymal stem cells (MSCs). This method of stem cell administration may allow for the longer viability and retention around damaged tissue by circumventing the initial immune response. A model of ischemia reperfusion injury was induced in mice accompanied by the delivery of mesenchymal stem cells (3.0 x 105), stably expressing the reporter gene firefly luciferase under the constitutively expressed CMV promoter, intramyocardially 7 days later. The animals were imaged via ultrasound and bioluminescent imaging for confirmation of injury and injection of cells, respectively. Importantly, there was no added complication rate when performing this two-procedure approach for SC delivery. This method of stem cell administration, collectively with the utilization of state-of-the-art reporter genes, may allow for the in vivo study of viability and retention of transplanted SCs in a situation of chronic ischemia commonly seen clinically, while also circumventing the initial pro-inflammatory response. In summary, we established a protocol for the delayed delivery of stem cells into the myocardium, which can be used as a potential new approach in promoting regeneration of the damaged tissue.

Stems cells are continuously investigated as potential treatments for individuals with myocardial damage, however, their decreased viability and retention within injured tissue can impact their long-term efficacy. In this manuscript we describe an alternative method for stem cell delivery in a murine model of ischemia reperfusion injury.

在穆林模型中缺血再灌注损伤后干细胞的心肌内传递延迟

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

温缺血/再灌注损伤的大鼠肺移植模型:改善结局的优化

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...