对塔基博士和 m. Funamoto 博士的技术支持给予了赞赏。

在实验之前, 所有的老鼠都应该得到一个星期的适应。所有动物的外科手术都应按照《动物护理和使用指南》 (www.nap.edu/catalog/5140.html) 或其他适当的道德准则进行。议定书应由动物福利委员会在适当的机构批准, 然后再继续。所有后续的程序必须在无菌条件下执行。
1. 准备体外循环电路
注: 佩戴个人防护用品, 包括手套、眼镜、清洁大衣或一次性长袍。
<ol><li>体外循环电路的设置<ol>
<li>将聚氯乙烯管与静脉储层连接, 预先设计为体外循环电路, 以及改良的新生儿膜肺, 如图 1所示。确保所有连接紧密, 不要漏水。</li>
		<li>根据制造商的协议, 将体外循环电路设置到滚筒泵装置。</li>
		<li>将滚筒泵保持在高度可调节的工作台上, 并将工作台的高度调整到实验台下面的10厘米处。</li>
	</ol>
</li>
	<li>体外循环电路的启动<ol>
<li>将12毫升羟乙基淀粉溶液与0.1 毫升肝素和0.5 毫升7% 碳酸氢钠溶液混合在一起启动体外循环电路。</li>
		<li>以11毫升的启动溶液为主要电路, 用泵辊轻轻旋转。把18口径的排气针放进储气库中。</li>
		<li>打膜肺几次到 deair, 与倾斜的肺。空气必须完全遮蔽, 以避免空气栓塞和氧合不足。在电路启动过程中, 通过设置在储层上的电热灯加热电路。</li>
	</ol>
</li>
</ol>2. 体外循环前的程序
注: 使用前应用70% 酒精或季铵盐化合物消毒手术现场和器械。
<ol><li>麻醉和动物设置<ol>
<li>麻醉一只在蒸发器中吸入3.0% 异氟醚混合空气的老鼠。把老鼠放在工作架上, 然后将16口径的套管插管到气管里。按照您当地的动物护理指南, 关于镇痛剂量和频率 (如: 丁丙诺啡0.005 毫克/千克) 注意: 老鼠必须在深度麻醉和失去反射。呼吸应该是有节奏的, 但不能被逮捕。</li>
		<li>将老鼠转到装有电热垫的手术台上。启动机械通气, 8 毫升/千克的潮汐量, 呼吸速率为70循环/分钟, 30% 的灵感氧分数监测的氧气传感器。</li>
		<li>用 1.5-2.0% 异氟醚维持麻醉, 并在体外循环启动时增加对氯胺酮/甲苯噻嗪的管理。</li>
		<li>用直肠探头监测直肠温度。通过调整热垫的温度并将电路置于散热灯上, 保持常温37摄氏度的体温。</li>
		<li>将大鼠仰卧位, 用针固定四肢。通过将心电图电极针设置为双侧肩和左腹, 对心率进行监测。放一条湿润的纱布或在眼睛上涂抹眼药膏以防止干燥。</li>
	</ol>
</li>
	<li>插管<ol>
<li>通过 sprayingby 喷雾70% 乙醇或另一种防腐溶液对全身表面进行消毒后, 在两侧腹股沟区和右颈部区域剃须剃刀毛。局部麻醉药 (如利多卡因) 应在进行皮肤切口前使用。注: 作为一种替代方法, 可使用切口部位的外科擦洗, 而不是全身喷雾70% 乙醇, 以避免体温下降。</li>
		<li>用剪刀切开皮肤 (大约5毫米) 在双侧腹股沟区域和右颈部区域, 并坦率地解剖组织以暴露右侧的主股动脉。从附近的静脉和神经仔细地分离动脉。结扎在股动脉末端由4-0 丝和紧张暴露。</li>
		<li>用微剪刀垂直向动脉切开右股动脉壁 (约1毫米), 并仔细 cannulate 24 口径静脉导管, 从切口到深度1厘米以监测全身动脉。动脉血中气体分压的压力和分析。</li>
		<li>从导管中管理肝素钠 (500 磅/千克)。</li>
		<li>按照步骤2.2.2 和 2.2.3 cannulate 24 口径静脉导管进入左股动脉, 作为体外循环回路的动脉输液线。</li>
		<li>将17口径的多孔 angiocatheter 插入右颈内静脉, 并将其推进右心房和下腔静脉。不要用力推导管, 因为容器很容易折断。将导管连接到体外循环回路进行静脉引流。</li>
		<li>用湿纱布覆盖每个空心区域以避免污染。</li>
	</ol>
</li>
</ol>3. 体外循环期间的程序
<ol><li>在体外循环期间, 将100% 氧气气体送到肺, 0.8 升/分, 将呼吸率降低到30周/分钟。动脉氧分压需要从200到 400 mmHg。</li>
	<li>在体外循环开始时, 将热垫的温度设置提高到最大值42摄氏度, 以减轻体外循环后体温的立即下降。当体温恢复到36摄氏度时, 将设定温度调整到37摄氏度。</li>
	<li>小心地启动体外循环流程, 密切关注水库中血液的体积。空的水库可能导致空气栓塞。如果储层中的血液体积减少, 通过调整工作台高度或改变引流导管的位置来降低泵流量。不要重新定位静脉引流导管, 这可能容易导致右心房穿孔和/或心律失常。</li>
	<li>增加并保持泵的流量为100毫升/千克/分钟, 而平均血压维持在 70 mmHg。当保持适当的血压时, 水库中至少有1毫升的小容积是可以接受的。如果水库中的血液少于1毫升, 则可能导致器官发生空气栓塞。</li>
	<li>如果血压不稳定, 增加2-3 毫升的启动方案的电路 (它可能导致贫血后, 体外循环)。</li>
</ol>4. 体外循环后的程序
<ol><li>钳住静脉引流管, 并将其从电路中取出。将血液中的剩余血逐渐注入动脉以维持血压。</li>
	<li>将呼吸率提高到70周/分钟。</li>
	<li>取出静脉引流导管和左动脉导管, 然后结扎近端和远端部位的血管。</li>
	<li>在体外循环结束后, 从右股动脉取出60分钟的动脉线。</li>
	<li>用生理盐水清洗伤口, 缝合伤口。</li>
	<li>检查动物的自发性呼吸, 结束麻醉, extubate 气管管。</li>
	<li>管理加热不育的等渗液, 并使用热垫和电热灯保持动物的温暖。经常检查动物的病情, 直到从麻醉中恢复。必要时提供呼吸支持。注意: 一旦动物开始走动, 热源应该从笼子的一部分移走, 让动物选择温暖或凉爽的一面。</li>
	<li>使动物远离其他动物的公司, 直到呼吸完全恢复。在完全恢复之前, 不要将动物退回公司。</li>
	<li>检查从麻醉恢复后的食物和水摄入量, 并提供适当的营养支持。管理止痛药, 检查是否有不适或疼痛的迹象。</li>
</ol>

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所有作者在商业支持方面没有什么可透露的。

在大鼠体外循环模型中, 体外循环后, 炎症细胞因子和 HMGB-1 的血清和肺表达水平是调节炎症反应的关键转录因子, 显著增加。以往的临床研究表明, 在心血管手术患者中, HMGB-1 水平的血清分泌增加了11, 体外循环期间血清 HMGB-1 水平与更严重的全身炎症反应综合征有关。体外循环后肺氧合功能障碍12。此外, 血清 HMGB-1 水平是一种独立的生物标志物, 用于预测重症肺?...

在 1953年, 约翰·小长臂猿成功地使用了 CPB1进行了第一次心脏手术, 随后它成为心血管外科的基本形态。虽然这些技术和设备已大为完善, 但与体外循环有关的多脏器并发症仍会损害心血管手术的结果, 并可能影响术后发病率和死亡率2。与体外循环有关的器官损害是由免疫系统活化和促炎细胞因子的合成、赞美活化和氧自由基的产生而引起的2。然而, 它的病理生理学尚未完全阐明。
动物模型综述的临床应用, 使对体外循环期间和之后的病理生理过程的澄清;这可以促进临床前研究, 以制定战略, 以避免这些并发症。自从鲍佩维奇et al。第一次报告了鼠体外循环模型在 1967年3, 大鼠体外循环模型已经完善, 并已逐渐取代大型动物模型, 由于更大的成本效益, 方便的实验过程, 和大量的测试方法在遗传和蛋白质水平。此外, 近亲繁殖的大鼠可以遗传相同, 减少可能的生物偏见。
Fabre et al。首先建立了恢复模型, 允许对 CPB 的长期多器官后遗症进行研究4。这种简单的生存模式的优点是灵活性 (体外循环流量和持续时间), 稳定的生命条件, 和重现性的系统性炎症。大鼠体外循环模型已成为研究预防体外循环5中多脏器损伤的治疗策略的关键, 最近开发了各种模拟体外循环临床情况的模型。De 兰格et al。建立了心脏骤停模型, 可用于描述与心肌损伤有关的酶、遗传和组织学反应7。彼得斯et al。采用小型化体外循环模型进行心肌梗死和控制再灌注, 通过局灶性缺血再灌注损伤分析心脏功能障碍8。Jungwirth et al。首先建立了深低温循环骤停 (DHCA) 模型, 它可以阐明 DHCA 的全球缺血再灌注损伤, 支持潜在神经保护策略6。研究使用 DHCA 调查体温过低, 再灌注和/或溶血触发信号事件的影响9。深低温可能会影响各种酶和通路的活化和失活, 而机制仍然未知的10。另一方面, 心脏骤停模型或心脏缺血模型必须用于研究缺血再灌注心脏损伤。这些不同的大鼠体外循环模型, 高度重述人类体外循环可能揭示与体外循环有关的病理过程, 并有助于缓解与体外循环相关的并发症。
该协议说明了一个简单的体外循环模型, 没有输血或强心剂剂的大鼠。该模型可用于研究体外循环的长期多脏器后遗症。

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a recovery cardiopulmonary bypass model without transfusion or inotropic agents in rats

体外循环 (CPB) 在心血管手术中是必不可少的。尽管体外循环技术和设备的戏剧性改良, 多脏器并发症与长期体外循环仍然影响心血管手术的结果, 并可能恶化的术后发病率和死亡率。动物模型综述的临床应用可以澄清在体外循环期间发生的病理生理过程, 并促进临床前期研究, 以制定预防这些并发症的战略。大鼠体外循环模型是有利的, 因为他们更大的成本效益, 方便的实验过程, 丰富的测试方法在基因或蛋白质水平, 和遗传一致性。它们可用于调查免疫系统激活和促炎性细胞因子的合成, 赞美活化和氧自由基的产生。大鼠模型已被提炼, 并逐渐取代了大型动物模型。在这里, 我们描述一个简单的体外循环模型, 没有输血和/或强心剂剂的老鼠。这种恢复模式允许研究体外循环的长期多器官后遗症。

图 1显示整个 CPB 电路。该模型中的生理变量显示在图 2中, 包括直肠温度、平均动脉血压和心率。图 3显示了体外循环期间的动脉血气分析, 包括动脉氧的分压, 动脉二氧化碳的分压, 红细胞积血, 碱过量, 血清钾的表达, 氢的电位。在整个手术过程中, 心率和平均动脉压力是稳定的。由于吸血量引起的血液稀?...

Watch this Scientific Journal Video about A Recovery Cardiopulmonary Bypass Model Without Transfusion or Inotropic Agents in Rats at JoVE.com

A Recovery Cardiopulmonary Bypass Model Without Transfusion or Inotropic Agents in Rats

无输血或强心剂剂的大鼠心肺旁路移植模型

在这里, 我们提出一个协议来描述一个简单的恢复心肺旁路模型没有输血或强心剂剂的大鼠。该模型可用于体外循环术后长期多脏器后遗症的研究。

Cardiopulmonary bypass (CPB) is indispensable in cardiovascular surgery. Despite the dramatic refinement of CPB technique and devices, multi-organ ...

a-recovery-cardiopulmonary-bypass-model-without-transfusion-or

Universidad de Cartagena UDEC

Research

JoVE Journal

Immunology and Infection

7.8K 次观看.  Kyoto University. 在这里, 我们提出一个协议来描述一个简单的恢复心肺旁路模型没有输血或强心剂剂的大鼠。该模型可用于体外循环术后长期多脏器后遗症的研究。

视频: A Recovery Cardiopulmonary Bypass Model Without Transfusion or Inotropic Agents in Rats

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

小鼠体外循环模型: 一种新方法

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

科研

医学

Pre-clinical Model of Cardiac Donation after Circulatory Death

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after circulatory death (DCD) is a solution to address this limited availability, but due to a period of prolonged warm ischemia and the risk of tissue injury, its routine use in cardiac transplantation is seldom seen. In this manuscript we provide a detailed protocol closely mimicking current clinical practices in the context of DCD with continuous monitoring of heart function, allowing for the evaluation of novel cardioprotective strategies and interventions to decrease ischemia-reperfusion injury.
In this model, the DCD protocol is initiated in anesthetized Lewis rats by stopping ventilation to induce circulatory death. When systolic blood pressure drops below 30 mmHg, the warm ischemic time is initiated. After a pre-set warm ischemic period, hearts are flushed with a normothermic cardioplegic solution, procured, and mounted onto a Langendorff ex vivo heart perfusion system. Following 10 min of initial reperfusion and stabilization, cardiac reconditioning is continuously evaluated for 60 min using intraventricular pressure monitoring. A heart injury is assessed by measuring cardiac troponin T and the infarct size is quantified by histological staining. The warm ischemic time can be modulated and tailored to develop the desired amount of structural and functional damage. This simple protocol allows for the evaluation of different cardioprotective conditioning strategies introduced at the moment of cardioplegia, initial reperfusion and/or during ex vivo perfusion. Findings obtained from this protocol can be reproduced in large models, facilitating clinical translation.

This protocol shows a simple and flexible approach for the evaluation of new conditioning agents or strategies to increase the feasibility of cardiac donation after circulatory death.

循环死亡后心脏捐献的临床前模型

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after ...

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 ...

A New Model of Heart Failure Post-Myocardial Infarction in the Rat

Ligation of the left anterior descending (LAD) coronary artery has been widely used to establish the rat model of heart failure (HF) post myocardial infarction (MI). However, the disadvantages of this model include high mortality rate after ligation and larger variations both in the infarct size and the degree of impaired cardiac function. In addition, a ventilator or exteriorization of the heart is indispensable for the previous models, which complicates the procedure during the ligation. In this study, we developed a reliable and reproducible model without the ventilator or exteriorization of the heart by ligating the LAD coronary artery. Four weeks after the procedure, we found that the serum concentrations of CK-MB, NT-proBNP, and Renin, which were used to assist diagnoses of MI and HF, were significantly higher in the MI group compared to the sham group. In contrast, the value of left ventricle ejection fraction (LVEF) in the MI group was obviously less than in the sham group. Furthermore, the infarct size and cardiac fibrosis area were individually confirmed and quantitatively analyzed by TTC staining and Masson's trichrome staining. Smaller variations were found in either infarct size or fibrosis area in the MI group, which helped to develop a reliable and reproducible model of HF post-MI. This new model of HF post-MI in the rat is vital for studying the potential mechanisms of MI and HF. This new method can be used to develop the new drug for treatment of MI and HF in rats by using pharmacological strategies.

We successfully developed a reliable and reproducible model of heart failure post-myocardial infarction in the rat without ventilation or exteriorization of the heart. This simplifies the procedure and benefits the further studies about the potential mechanisms behind the heart failure.

Ligation of the left anterior descending (LAD) coronary artery has been widely used to establish the rat model of heart failure (HF) post myocardial ...

Large-Animal Model of Donation after Circulatory Death and Normothermic Regional Perfusion for Cardiac Assessment

The increase in demand for cardiac transplantation throughout the years has fueled interest in donation after circulatory death (DCD) to expand the organ donor pool. However, the DCD process is associated with the risk of cardiac tissue injury due to the inevitable period of warm ischemia. Normothermic regional perfusion (NRP) allows for an in situ organ assessment, allowing the procurement of hearts determined to be viable. Here, we describe a clinically relevant large-animal model of DCD followed by NRP. Circulatory death is established in anesthetized pigs by stopping mechanical ventilation. After a preset warm ischemia period, an extracorporeal membrane oxygenator (ECMO) is used for a NRP period lasting at least 30 min. During this reperfusion period, the model allows the collection of various myocardial biopsies and blood samples for initial cardiac evaluation. Once NRP is weaned, biochemical, hemodynamic, and echocardiographic assessments of cardiac function and metabolism can be performed before organ procurement. This protocol closely simulates the clinical scenario previously described for DCD and NRP in heart transplantation and has the potential to facilitate studies aimed at decreasing ischemia-reperfusion injury and enhance cardiac functional preservation and recovery.

The protocol describes a large-animal (porcine) model of donation after circulatory death, followed by thoracoabdominal normothermic regional perfusion that closely simulates the clinical scenario in heart transplantation, and has the potential to facilitate therapeutic studies and strategies.

用于心脏评估的循环死亡后捐献和正常区域灌注的大型动物模型

The increase in demand for cardiac transplantation throughout the years has fueled interest in donation after circulatory death (DCD) to expand the organ ...

Rat Model of Normothermic Ex-Situ Perfused Heterotopic Heart Transplantation

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the number of heart failure patients waiting for transplantation is still increasing. The normothermic ex situ preservation technique has been established as a comparable method to the conventional static cold storage technique. The main advantage of this technique is that donor hearts can be preserved for up to 12 h in a physiologic condition. Moreover, this technique allows resuscitation of the donor hearts after circulatory death and applies required pharmacologic interventions to improve donor function after implantation. Numerous animal models have been established to improve normothermic ex situ preservation techniques and eliminate preservation-related complications. Although large animal models are easy to handle compared to small animal models, it is costly and challenging. We present a rat model of normothermic ex situ donor heart preservation followed by heterotopic abdominal transplantation. This model is relatively cheap and can be accomplished by a single experimenter.

Here, we present an assessment protocol of a heterotopically implanted heart after normothermic ex situ preservation in the rat model.

异位灌注异位心脏移植大鼠模型

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the ...

心脏骤停后动物模型体外心肺复苏

Pediatric Animal Model of Extracorporeal Cardiopulmonary Resuscitation After Prolonged Circulatory Arrest

Congenital heart disease (CHD) is the most prevalent congenital malformation, with about one million births impacted worldwide per year. Comprehensive investigation of this disease requires appropriate and validated animal models. Piglets are commonly used for translational research due to their analogous anatomy and physiology. This work aimed to describe and validate a neonatal piglet model of cardiopulmonary bypass (CPB) with circulatory and cardiac arrest (CA) as a tool for studying severe brain damage and other complications of cardiac surgery. In addition to including a list of materials, this work provides a roadmap for other investigators to plan and execute this protocol. After experienced practitioners performed several trials, the representative results of the model demonstrated a 92% success rate, with failures attributed to small piglet size and variant vessel anatomy. Furthermore, the model allowed practitioners to select from a wide variety of experimental conditions, including varying times in CA, temperature alterations, and pharmacologic interventions. In summary, this method uses materials readily available in most hospital settings, is reliable and reproducible, and can be widely employed to enhance translational research in children undergoing heart surgery.

作者聚焦：使用新生儿仔猪模型模拟儿科心脏手术

This protocol describes a neonatal porcine model of cardiopulmonary bypass (CPB), with circulatory and cardiac arrest as a tool for studying severe brain damage and other complications secondary to CPB.

长时间循环停止后体外心肺复苏的小儿动物模型

Congenital heart disease (CHD) is the most prevalent congenital malformation, with about one million births impacted worldwide per year. Comprehensive ...

Modified Heterotopic Abdominal Heart Transplantation and a Novel Aortic Regurgitation Model in Rats

Over the past 50 years, many researchers have reported heterotopic abdominal heart transplantation in mice and rats, with some variations in the surgical technique. Modifying the transplantation procedure to strengthen the myocardial protection could prolong the ischemia time while preserving the donor&#39;s cardiac function. This technique&#39;s key points are as follows: transecting the donor&#39;s abdominal aorta before harvesting to unload the donor&#39;s heart; perfusing the donor&#39;s coronary arteries with a cold cardioplegic solution; and topical cooling of the donor&#39;s heart during the anastomosis procedure. Consequently, since this procedure prolongs the acceptable ischemia time, beginners can easily perform it and achieve a high success rate.
Moreover, a new aortic regurgitation (AR) model was established in this work using a technique different from the existing one,&#160;which is created by inserting a catheter from the right carotid artery and puncturing the native aortic valve under continuous echocardiographic guidance. A heterotopic abdominal heart transplantation was performed using the novel AR model. In the protocol, after the donor&#39;s heart is harvested, a stiff guidewire is inserted into the donor&#39;s brachiocephalic artery and advanced toward the aortic root. The aortic valve is punctured by pushing the guidewire further even after the resistance is felt, thus inducing AR. It is easier to damage the aortic valve using this method than with the procedure described in the conventional AR model. Additionally, this novel AR model does not contribute to the recipient&#39;s circulation; therefore, this method is expected to produce a more severe AR model than the conventional procedure.

This study demonstrates a reproducible heterotopic abdominal heart transplantation technique in rats that beginners can learn and perform. Additionally, a novel aortic regurgitation model in rats is generated by performing heterotopic abdominal heart transplantation and damaging the donor's aortic valve using a guidewire after harvesting.

改良异位腹心移植及新型大鼠主动脉瓣反流模型

Over the past 50 years, many researchers have reported heterotopic abdominal heart transplantation in mice and rats, with some variations in the surgical ...

无输血或强心剂剂的大鼠心肺旁路移植模型

摘要

探索更多视频

此视频中的章节

小鼠体外循环模型: 一种新方法

循环死亡后心脏捐献的临床前模型

温缺血/再灌注损伤的大鼠肺移植模型:改善结局的优化

A New Model of Heart Failure Post-Myocardial Infarction in the Rat

用于心脏评估的循环死亡后捐献和正常区域灌注的大型动物模型

异位灌注异位心脏移植大鼠模型

长时间循环停止后体外心肺复苏的小儿动物模型

长时间循环停止后体外心肺复苏的小儿动物模型

长时间循环停止后体外心肺复苏的小儿动物模型

改良异位腹心移植及新型大鼠主动脉瓣反流模型