作者没有致谢。

本议定书的所有实验均由瑞士苏黎世的兽医当局批准 (编号 60/2014)。此外, 所有实验步骤都严格遵照瑞士医学科学院 (sam) 的《动物实验指南》和欧洲实验动物科学协会联合会 (FELASA) 的指导方针进行。.
1. 畜牧, 手术室设备和器械, 麻醉
<ol><li>在标准无病原体条件下, 在12/12 小时的光照/暗循环下, 在通风笼中保持雄性大鼠体重250-300 克。在22±1° c 的环境温度下, 让动物免费获得食物和水。</li>
	<li>诱导麻醉在一个盒子, 其中异氟醚的冲洗与 5% 三十年代 (图 1A), 其次是异氟醚浓度 3%, 直到该动物在深麻醉。用脚趾捏反射来确认麻醉的程度。</li>
	<li>在手术过程中使用丁丙诺啡 (0.01 毫克/千克) 提供镇痛, 在术后0.02 毫克/千克, 每12小时为48小时。</li>
	<li>将动物转移到工作场所, 在那里他们自发地呼吸暴露在氧气中的 1.0-2.5% 异氟醚 (流量: 600 毫升/分钟) 的气体混合物。</li>
	<li>闭上动物的眼睛, 用药膏保护眼睛。注射5毫升生理盐水皮下注射, 其中一半在腹部两侧, 其次是0.1 毫克阿托品。</li>
	<li>在显微外科手术室使用手术显微镜进行外科治疗。</li>
	<li>使用由乳胶膜 (图 1B) 组成的透气面罩将动物置于挥发性麻醉下, 从而推动老鼠的鼻子。用胶带固定四肢 (图 1B)。</li>
	<li>在一个侧面表上放置无菌的手术器械: 盐水和控告浸泡的海绵 (图 2A), 腹壁器 (图 2B) 以及3-0 缝线, 用于收缩腹壁和剑、剪刀、Adson镊子, 细提示 microforceps 直和弯曲和预6-0 丝领带结扎程序 (图 2C)。</li>
	<li>用 non-sticking 双极 microforceps 对肝实质进行横断。用 SH-1 针和 5-0 Maxon (图 2D) 关闭腹壁和皮肤, 使用3-0 丝缝线。用双极钳止血。</li>
</ol>2. 手术开始
<ol><li>用小动物的毛剪从剑到生殖器, 用中线的2厘米的区域来刮毛动物的腹部。彻底去除头发。</li>
	<li>用控告浸泡过的海绵将这个区域消毒3次。</li>
	<li>用手术刀为皮肤做中线切口, 然后用手术剪刀打开腹部。</li>
	<li>用3-0 丝缝线 (图 3A) 收回右、左腹壁和剑。</li>
	<li>用自制的小丝器 (图 3B) 从大鼠的十二指肠韧带中收回胃、结肠和小肠。</li>
</ol>3. 门静脉结扎术 (PVL)
<ol><li>通过胃和小肠的侧向收缩来进入门静脉及其分支 (图 3B)。</li>
	<li>直接解剖门静脉, 用 microforceps 的腹膜, 慢慢剥去内侧。</li>
	<li>
解剖门静脉分支 (图 3C) 按以下顺序排列: (1) 右后支, (2) 左侧和左中间分枝, 和 (3) 尾状枝。
	<ol>
<li>首先, 剥离腹膜与小, 不太积极的动作, 以避免撕裂小动脉分支到肝裂片, 运行直接在腹膜覆盖。</li>
		<li>一旦主门静脉被裸露, 包围的树枝缓慢向前推和传播运动, 然后拉在各自的门静脉分支周围的1厘米6-0 丝领带, 并结扎它。</li>
	</ol>
</li>
	<li>
结扎右后叶分支。(图 3C)右后支的技术难点在于右后叶的动脉贯穿右后门静脉分支。
	<ol>
<li>将动脉 cranially 与腹膜结合, 使右门静脉分支与弯曲的 microforceps。</li>
		<li>结扎右后叶分支与预6-0 丝领带。一旦门静脉结扎, 右后叶明显变苍白。</li>
		<li>避免在盘旋时推挤抵抗阻力, 以防止撕裂动脉或门静脉。在出血的情况下, 使用无菌棉签至少一分钟的压力, 然后重新评估。如果流血不停止, 牺牲动物。</li>
	</ol>
</li>
	<li>
结扎左外侧 (i) 和左中位叶 (爱我的生活) 分支。这些树枝有一个共同的树干。(图 3C)。左中位裂片分支从门静脉内出现在微光的实质中。
	<ol>
<li>结扎的门静脉导致我和爱我的生活分支一起使用预6-0 丝领带。</li>
		<li>避免受伤的动脉导致我和爱我的生命, 运行在腹膜层。只要腹膜仔细剥去, 它们就完好无损。一旦树枝结扎, 我的生命就会变得苍白。</li>
	</ol>
</li>
	<li>
结扎尾状叶分支 (图 3C)。门静脉导致稍微分开的尾状裂片在大鼠从主要门静脉, 上部和内侧到从主要门静脉 (不严密地下象在人或更大的动物) 的右叶分支的起飞。
	<ol>
<li>解剖它和包围它通过解除腹膜加胆管和动脉从主门静脉。</li>
		<li>包围尾状门静脉分支与来自侧面的弯曲 microforceps。当尾状叶分支被6-0 丝结扎后, 两尾状叶明显变苍白。</li>
	</ol>
</li>
	<li>捆绑这三门静脉分支导致专属门静脉灌注右中位叶 (RML), 这是可见的一个明确的分界线之间的权利和左中位叶。</li>
</ol>4. 门静脉结扎并横断 (PVL + T)
<ol><li>执行沿分界线 RML 和爱我的生活后, PVL (图 3D)。</li>
	<li>使用一个 non-sticking 双极银钳与一个非常精细的神经外科提示 precauterize 肝组织创建一个1毫米烧灼条沿分界线。</li>
	<li>用剪刀小心地切开 precauterized 组织。要正确评估横断面的深度是相当困难的。目标是得到尽可能接近腔静脉, 在大鼠中运行 intrahepatically。腔静脉的损伤伴随于广泛的流血, 可能通常不被停止。</li>
	<li>在受伤的情况下使用棉签轻轻按压, 但如果出血不停止在 3-4 分钟内, 牺牲的动物。使用缝线修复腔静脉的尝试通常不成功。大鼠对空气栓塞非常敏感, 有时可能发生, 导致心脏骤停。</li>
</ol>5. 门静脉压力和容积流量的术中测量
<ol><li>通过与压力监测器连接的 G30 针直接插管来评估门静脉压力 (图 4A)。</li>
	<li>使用2毫米容积流探头横向连接到流量装置上, 直接测量入口容积流量超过2分钟 (图 4B)。在机器显示屏上, 只记录数据, 测量质量稳定。</li>
	<li>在定义相应程序的一组步骤之后, 分别评估门静脉流量和压力,即三门静脉分支的结扎或三门静脉分支的结扎, 再加上执行实质横断在 RML 和爱之间我的生活。</li>
</ol>6. 手术的最后步骤
<ol><li>在关闭前, 确保小肠放置在腹腔内。</li>
	<li>用 SH-1 针缝合腹膜和腹壁3-0 丝。</li>
	<li>最后缝合皮肤使用 Maxon 5-0。</li>
	<li>再给皮肤消毒。</li>
	<li>再次应用5毫升生理盐水皮下注射到腹部的侧皮组织。</li>
</ol>7. 小动物 CT 对大鼠肝容量的影响
<ol><li>24小时前的第一次 CT 成像注入200μ l 的造影剂进入尾部静脉通过 G26 静脉导管。</li>
	<li>用微 CT 扫描老鼠。将动物置于有机玻璃盒中以诱导全身麻醉 (三十年代第一次冲洗 5%, 其次是 2-3% 的异氟醚浓度, 直到该动物被麻醉并通过油管系统进行扫描。为了获得最佳的图像质量, 请将 CT 扫描与动物的呼吸同步。</li>
	<li>输出数字成像和通信的医学 (DICOM) 文件和分析他们使用公共领域成像平台 OsiriX 8.0。使用 "闭合多边形" 工具根据肝脏中纳米粒子对比度的 radiodensitiy 绘制耳垂边框, 并使用感兴趣区域 (ROI) 菜单计算部分肝脏容量 (请参见 Schadde et al.中的图 2A17)</li>
	<li>在标准模型中, 允许肝脏生长超过3天, 并获得每日 CT 扫描, 以评估体积增加。</li>
</ol>

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P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+Liver+Function+Using+(99m)Tc-Mebrofenin+Hepatobiliary+Scintigraphy+in+ALPPS+(Associating+Liver+Partition+and+Portal+Vein+Ligation+for+Staged+Hepatectomy).">Assessment of Liver Function Using (99m)Tc-Mebrofenin Hepatobiliary Scintigraphy in ALPPS (Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy).</a> Case Rep Gastroenterol. 9 (3), 353-360 (2015).</li><li>Truant, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drop+of+Total+Liver+Function+in+the+Interstages+of+the+New+Associating+Liver+Partition+and+Portal+Vein+Ligation+for+Staged+Hepatectomy+Technique:+Analysis+of+the+&amp;#34;Auxiliary+Liver&amp;#34;+by+HIDA+Scintigraphy.">Drop of Total Liver Function in the Interstages of the New Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy Technique: Analysis of the &amp;#34;Auxiliary Liver&amp;#34; by HIDA Scintigraphy.</a> Ann Surg. 263 (3), e33-e34 (2016).</li></ol>

作者没有什么可透露的。

本议定书提出了一个动物模型的 ALPPS 与其快速肥大诱导的 PVL + T, 大约双打增加3天内的体积比单独 PVL。17右中肝叶被用作生长中的肝叶的模型, 因为中间的肝叶是由两个独立的门静脉向其左侧和右侧提供的一个连续的实质质量, 如图 1所示, 最近已发布的工作。17与其他报表相比, 该模型具有一些优点。从解剖学上来说, 选择中叶最能代表人的肝脏作为一个连续的实质质量, 允许横断, 从而闭塞的侧枝流。门静脉的分支到左中位裂片 (爱我的生活) 在左侧面裂片 (微光) 之内允许联合结扎爱我的生活和我。
有各种各样的老鼠和一个小鼠模型为 ALPPS 开发和描述。18许多老鼠模型19,20和唯一的鼠标模型15描述需要肝脏大量减少。大体积复位可独立触发增强肥大。这与我们的17和类似的大鼠模型形成了对比。14,21,22,23小鼠模型的优点可能是使用转基因动物。或者对这些小动物模型, 猪模型被开发了,16使用相似的外科技术象在人和导致相似的成长率。尽管事实上, 这可能是最接近人类的外科手术, 小动物模型可能仍然是非常重要的, 因为它们是更快, 更容易执行, 允许更大数量的动物每组和较低的成本。
从生理学上来说, 老鼠模型可以比老鼠模型更容易地研究血流、抵押和氧合。这项研究是第一次使用同样的方法在人类临床上使用, 以评估肝脏体积的变化, 即 CT 容量。与其他一些研究不同的是, 它不包括一个对照组 (例如在 ALPPS24的猪模型中), 但却假设 ALPPS 生长动力学, 我们通过比较 PVL 和 PVL 来验证肥大。而在 PVL 下 RML 在3天内加倍, PVL + T 增加肥大显着。这加速肥大后3天由增加的横断面恰恰反映了人类的情况, 其中46% 体积增加已报告在一个大的 meta 分析 PVE 和86% 增加在 ALPPS 后10天在第一个大型 ALLPS 注册表报告202例。25大鼠程序 PVL + T 和人 ALPPS 两种方法在各自的时间框架内实现的肥大的数量加倍。
本协议中的关键步骤包括 PVL 门静脉分支的解剖和 PVL 中位叶的横断。在门静脉分支的解剖过程中, 重要的是要阻断覆盖整个十二指肠韧带的薄层腹膜, 而不是将器械推向阻力。在门静脉的意外撕裂可能会停止压力使用棉签单独, 但可能是无法挽回的, 需要牺牲的动物。一般来说, 必须强调的是, 动物失血量超过估计的10% 的动物的血容量, 应排除在研究之外, 因为这可能改变研究结果和造成不必要的痛苦的动物。
通过细银双极钳和足够的盐水滴烧灼, 然后用剪刀简单切割烧灼组织, 可以实现正中叶的横断。在遇到大鼠肝脏内的腔静脉时, 应停止横断, 以避免大出血和空气栓塞进入腔静脉。空气进入肺循环导致大鼠心脏骤停。
本议定书的故障排除可能包括所有的修改, 导致增加动物的生理压力, 如低血压诱发的 oversedation, 体温过低, 增加失血和太长的手术时间。这项协议很可能被修改为其他啮齿动物种类象老鼠。我们成功地完成了在 C57BL/6 小鼠中描述的技术 (没有显示数据)。
该协议的一个局限是本议定书的独家使用, 研究快速与慢速肥大的机制;因此, 第二阶段的 "ALPPS" 程序, 切除所有 deportalized 肝组织除肥大肝脏, 没有在本议定书中描述。然而, 这种切除术可以很容易地达到以下标准技术肝切除啮齿动物, 使用缝合结扎, 然后用剪刀切除肝裂片。
总的来说, 目前的模型允许实验阐明快速肝肥大的机制和测试药物和干预。使用这个模型, 最近表明, RML 的快速肥大也可以诱导的门静脉结扎结合缺氧信号使用脯羟化酶抑制剂, 提示缺氧信号我的发挥重要作用肝生长动力学调控。像脯羟化酶抑制剂这样的药物可能加速肝脏再生, 并应进一步试验。17,26该模型的未来应用可能是, 常规和新型化疗药物的作用可能被测试, 它们对加速肝脏再生的影响可以进一步阐明。该模型还提供了研究肝功能的机会, 例如哚绿色 (组织) 或肝胆乙酸酸 (飞) 显像在大鼠, 因为最终缓慢和快速的体积变化将不得不放在背景肝功能的功能性肝再生比肝体积更重要。27,28对 ALPPS 患者的评估, 其中有29个与飞的影像,3031到目前为止只在回顾性队列研究中测试过, 但这些仍然是回答问题的非常重要的工具。在快速肝再生中, 体积或功能的变化是否相似或不同。
总之, 我们提出了一个良好的特点和标准化模式的快速和缓慢的肝脏再生, 这将允许未来的研究再生肝外科。

肝脏残余物的大小限制了肝脏肿瘤的可切除性。1一般来说, 当少于25% 的肝脏组织被留下时, 患者由于缺乏新陈代谢的作用为整个有机体 ("太小为大小综合症状), 增加死亡的风险从急性肝功能衰竭。2此 post-hepatectomy 肝衰竭是肝切除术后最具破坏性的并发症。因此, 临床医生试图通过操纵门静脉的流动, 在肝切除前诱导肝再生。3发现, 一旦门静脉被阻塞, 门静脉流的剩余部分开始以较慢的速度生长, 从而可以增加到60% 的大小。4手术结扎5或介入性门静脉闭塞均已临床建立。4肝脏体积和功能的增加是可靠的, 但肝门闭塞后肝脏的生长率只有 1/5, 与部分肝切除后残肝的生长情况比较。6
肝脏生长所需的时间是几周到几个月, 即使肝脏可以在切除后以更快的速度再生。因此, 肝脏是唯一的器官, 生长回正常功能后, 删除的一部分。7一个新的程序, 诱导肝脏再生的速度与部分 hepactectomy 后, 由一组医生发现, 增加横断之间的闭塞和 non-occluded 部分肝脏诱导肝肥大与肝切除后相同的生长速率, 但在切除前。9该过程在未来一周内启动快速肥大的80% 的肝脏残余物, 允许切除广泛的, 主要不能切除的, 肝脏肿瘤在一周内。这一过程被称为 "结合肝分割和门静脉结扎术分期切除 = ALPPS", 并迅速风靡世界各地。10多个报告支持新技术实现的边缘切除肝肿瘤可切除性的扩展,11而复杂的外科手术也因其高并发症率而受到批评。12,13
自 ALPPS 于2012年出版以来, 一直试图发展一种啮齿动物和大型动物模型, 以使其能够更好地鉴定和了解这些机制, 并测试药物对动物肝脏组织的不同生长速率。开发的第一个动物模型是老鼠模型。在该模型中, 左、右正中叶实质横断后快速肥大, 右中叶加速再生。14稍后在鼠标中引入了不同的模型。在这个模型中, 左外侧叶切除, 门静脉分支到肝脏的每一个肺叶, 除了左中位叶被捆绑。15同时, 还描述了猪 ALPPS 的大型动物模型。16
为研究肝门静脉血流变化和压力、肝脏组织灌注和氧合等生理机制, 大鼠模型优于小鼠 ALPPS 模型。老鼠模型的另一个优点是, 在大鼠模型中, 没有必要切除左外侧叶,15可能会污染肝切除与 ALPPS 的影响。大鼠模型的对比并不能降低肝细胞的质量。猪的模型使用右后叶作为生长中的叶, 但猪肝是高度叶的。因此, 在右后、右前叶之间的已经薄的组织桥上建立一个横断平面是很困难的。相比之下, 大鼠的中位叶由两部分组成, 分别由门静脉提供, 并且在两者之间使用显微外科技术可以很容易地形成实质横断平面。小动物计算机断层扫描 (CT) 和/或磁共振成像 (MRI) 的可用性使得门静脉结扎与门静脉结扎和附加横断之间的容积增长非常精确, 这一点很重要以验证任何快速肝肥大模型。
这里介绍的协议描述了用于容积验证的手术技术和程序, 以及门静脉结扎和门静脉结扎术后慢、快速肥大模型的生理特性,分别在大鼠。

<table><tbody><tr><td>Isoflurane, 250 mL bottles</td><td>Attane, Piramal, Mumbai, India</td><td>LDNI 22098</td><td>Standard vet. equipment</td></tr><tr><td>Tec-3 Isofluorane Vaporizer</td><td>Ohmeda, GE-Healthcare, Chicago, IL</td><td>not available anymore</td><td>Standard vet. equipment&amp;nbsp;</td></tr><tr><td>Buprenorphine (Temgesic)</td><td>Indivior, Baar, Switzerland</td><td>7680419310353</td><td>GTIN-number</td></tr><tr><td>Vitamine A ointment</td><td>Bausch&amp;amp;Lomp, Zug, Switzerland</td><td>7680223980247</td><td>GTIN-number</td></tr><tr><td>Atropine sulfate 0.5 mg/mL</td><td>Sintetica SA, Mendrisio, Switzerland</td><td>7680565330045</td><td>GTIN-number</td></tr><tr><td>Microsurgery microscope</td><td>Olympus, Tokio, Japan</td><td>SZX10</td><td>Standard vet. equipment</td></tr><tr><td>Betadine</td><td>Mundipharma, Basel, Switzerland</td><td>7680342821377</td><td>GTIN-number</td></tr><tr><td>Sponges</td><td>Carl Roth GmbH, Karlsruhe, Germany</td><td>NK83.1</td><td>Mini-sponges</td></tr><tr><td>Abdominal Wall retractors</td><td>N/A</td><td>N/A</td><td>Self-made from paper clips and Q-Tips</td></tr><tr><td>3-0 silk&amp;nbsp;</td><td>Ethicon, Sommerville, NJ</td><td>K872H</td><td>Standard surgical</td></tr><tr><td>Scissors&amp;nbsp;</td><td>World precision instruments (WPI), Sarasota, FL</td><td>503371</td><td>Standard microsurgical</td></tr><tr><td>Adson forceps</td><td>World precision instruments (WPI), Sarasota, FL</td><td>501244-G</td><td>Standard microsurgical</td></tr><tr><td>Fine tips microforceps</td><td>World precision instruments (WPI), Sarasota, FL</td><td>501976</td><td>Tips need to be polished regularly</td></tr><tr><td>Curved fine tips microforceps</td><td>World precision instruments (WPI), Sarasota, FL</td><td>504513</td><td>Essential to go around the portal vein branches&amp;nbsp;</td></tr><tr><td>6-0 LOOK black braided silk</td><td>Surgical Specalities Corporation, Wyomissing, PA&amp;nbsp;</td><td>SP114</td><td>Spool, precut prior to the procedure</td></tr><tr><td>2-0 silk sutures</td><td>Ethicon, Sommerville, NJ</td><td>K833</td><td>Standard surgical</td></tr><tr><td>5-0 maxon sutures</td><td>Covidien, Dublin, Ireland</td><td>6608-21</td><td>Standard surgical</td></tr><tr><td>Bipolar microforceps</td><td>Sutter, Freiburg, Germany</td><td>780148SGS</td><td>Essential for parenchymal transection</td></tr><tr><td>Q-tips small</td><td>Carl Roth GmbH, Karlsruhe, Germany</td><td>EH11.1</td><td>Standard surgical</td></tr><tr><td>Q-tips big</td><td>Carl Roth GmbH, Karlsruhe, Germany</td><td>XL54.1</td><td>Standard surgical</td></tr><tr><td>G30 needle&amp;nbsp;</td><td>Terumo, Tokyo, Japan</td><td>NN-3013R&amp;nbsp;</td><td>Standard anesthesia equipment</td></tr><tr><td>2 mm volume flow probe&amp;nbsp;</td><td>Transonic Systems, Ithaca, NY</td><td>MA-2PS</td><td>Smallest available probe for HAT-311 flow meter</td></tr><tr><td>Transonic flow meter</td><td>Transonic Systems, Ithaca, NY</td><td>HAT-311 Transsonic flow QC meter</td><td>One of the&amp;nbsp; first generation flow flow meters for surgery</td></tr><tr><td>ExiTron nano 12,000&amp;nbsp;</td><td>Miltenyi Biotech, Bergisch Gladbach, Germany</td><td>130-095-698</td><td>Nanomoloecular contrast medium that opacifies liver and spleen</td></tr><tr><td>G26 intravenous catheter</td><td>Becton Dickinson, Franklin Lakes, NJ</td><td>391349</td><td>Standard anesthesia equipment</td></tr><tr><td>Quantum FX MicroCT&amp;nbsp;</td><td>Perkin Elmer, Waltham, MA</td><td>N/A</td><td>Standard small animal CT scanner at the institute of physiology, University of Z&amp;uuml;rich</td></tr><tr><td>OsiriX 8.0</td><td>Pixmeo Sarl, Geneva, Switzerland</td><td>N/A</td><td>Public domain software : www.pixmeo.com</td></tr></tbody></table>

rat model of the associating liver partition and portal vein ligation for staged hepatectomy (alpps) procedure

最近的临床数据支持对原发性和转移性肝肿瘤采取积极的手术方法。对于某些适应症, 如结直肠癌肝转移, 肝切除后留下的肝组织数量已成为大、多肝肿瘤可切除性的主要限制因素。为了避免 post-hepatectomy 肝衰竭的严重并发症, 需要少量的功能性组织, 发病率和死亡率都很高。在肝外科手术中, 以门静脉栓塞的形式, 或以门静脉结扎术前几周的形式, 在肝脏切除术前, 诱导肝脏的生长变得更加成熟。最近, 肝脏再生是更广泛和快速的, 当实质横断被增加到门静脉结扎在第一阶段然后, 在仅一个星期等待之后, 切除在第二个阶段执行 (关联肝脏分割和门静脉结扎分期切除 = ALPPS)。ALPPS 在世界范围内迅速普及, 但因其围手术期死亡率高而备受诟病。这一程序引起的加速和广泛增长的机制还没有得到很好的理解。建立了动物模型, 探讨了 ALPPS 加速肝再生的生理和分子机制。该协议提出了一个大鼠模型, 允许机械探索加速再生。

两种不同的手术方法门静脉结扎 (PVL) 和 PVL 与横断 (PVL + T) 导致明显不同的生长动力学。PVL 在3天内会导致适度的体积增大, 而在 PVL + T 中可以看到更大的右中位叶 (RML) (图 5)。这可以通过每日容量来验证。RML 的体积大约在3天内的 PVL 倍, 而它在 PVL + T.17
流量测量在门静脉显露一个稳定的流程, 尽管减少的流动区域在 PLV 并且 PVL + T (参见图 3A在 Schadde et al.17). 这表明, 整个门静脉血流量是通过约26% 的前肝实质, 从而导致 "门户气力"。
门静脉内的压力测量显示, PVL 和 PVL 的平均门静脉压力从5柱到9柱的急性增加 (请参阅 Schadde et al.中的图 3B17). 这可能是门户气力的结果。单独横断面不会导致急性门脉压增高, 因为肝脏组织的体积不会减少。17重复测量表明, 24 h.17的压力增加保持稳定
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55895/55895fig1.jpg"> 图 1: 麻醉.(A) 在用异氟醚 (5%) 冲洗一盒以诱导动物麻醉后, 异氟醚被汽化 (600 毫升氧气/分钟) 用于全麻的维持。(B) 在操作显微镜下, 动物被固定在不育的操作表面上。<a href="http://cloudflare.jove.com/files/ftp_upload/55895/55895fig1large.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/55895/55895fig2.jpg"> 图 2: 无菌设备.(A) 用于曝光和缩回的无菌湿润海绵。(B) 连接到橡皮带的线拖斗。(C) 双极钳、茶煲剪刀;弯曲 microforceps, 直 microdissectors 用于解剖门静脉分支。(D) 较大的剪刀、针夹和针用于打开和关闭。<a href="http://cloudflare.jove.com/files/ftp_upload/55895/55895fig2large.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/55895/55895fig3.jpg"> 图 3: 解剖.(A) 腹部和剑的两侧经中线剖腹手术后缩回3-0 丝缝线。(B) 通过缩回胃, 小和大肠侧向大鼠的胃十二指肠韧带, 肝门可以暴露。(C) 在门静脉结扎 (PVL) 中, 门静脉分支到右叶 (1), 左外侧叶 (2) 和尾状裂片 (3) 结扎6-0 丝领带 (左)。在 PVL 与横断 (PVL + T), 另外, 缺血性分界线在右中位叶 (RML) 和左中位叶 (爱我的生活) 之间是断 (正确)。(D) 照片显示了爱我的生活和 RML (左) 之间清晰的分界线。这条线是遵循的横断面使用双极性 microforceps PVL + T (右)。<a href="http://cloudflare.jove.com/files/ftp_upload/55895/55895fig3large.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/55895/55895fig4.jpg"> 图 4: 术中测量门静脉压力和容积流量.(a) 在过程中, 将针式压力传感器插入暴露的门静脉。(B) 2 mm 容积流量探头用于测量主门静脉的容积流量。<a href="http://cloudflare.jove.com/files/ftp_upload/55895/55895fig4large.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/55895/55895fig5.jpg"> 图 5: 肝容量.肝脏在 PVL 以后增加更多在大小比 PVL 单独以后。该图显示了每日容量进行的数字成像和医学通信 (DICOM) 文件获得的大鼠微 CT 扫描仪和使用公共领域的成像平台。虽然在轴向图像上的差异是明显的, 在前一份报告17中执行的定量容量显示了生长动力学的显著差异。<a href="http://cloudflare.jove.com/files/ftp_upload/55895/55895fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure at JoVE.com

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

提出了联合肝分型和门静脉结扎术 (ALPPS) 诱导肝脏快速肥大切除交界性切除肝肿瘤的方法。该模型可以阐明参与快速肥大的机制, 并允许对促进或阻止再生加速的药物进行检测。

肝分型与门静脉结扎大鼠分期肝切除 (ALPPS) 手术模型的建立

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver ...

rat-model-associating-liver-partition-portal-vein-ligation-for-staged

Nanyang Technological University

Research

JoVE Journal

Medicine

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure

14.6K 次观看.  University of Zurich. 提出了联合肝分型和门静脉结扎术 (ALPPS) 诱导肝脏快速肥大切除交界性切除肝肿瘤的方法。该模型可以阐明参与快速肥大的机制, 并允许对促进或阻止再生加速的药物进行检测。

视频: Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

Cecal Ligation Puncture Procedure

Human sepsis is characterized by a set of systemic reactions in response to intensive and massive infection that failed to be locally contained by the host. Currently, sepsis ranks among the top ten causes of mortality in the USA intensive care units 1. During sepsis there are two established haemodynamic phases that may overlap. The initial phase (hyperdynamic) is defined as a massive production of proinflammatory cytokines and reactive oxygen species by macrophages and neutrophils that affects vascular permeability (leading to hypotension), cardiac function and induces metabolic changes culminating in tissue necrosis and organ failure. Consequently, the most common cause of mortality is acute kidney injury. The second phase (hypodynamic) is an anti-inflammatory process involving altered monocyte antigen presentation, decreased lymphocyte proliferation and function and increased apoptosis. This state known as immunosuppression or immune depression sharply increases the risk of nocosomial infections and ultimately, death. The mechanisms of these pathophysiological processes are not well characterized. Because both phases of sepsis may cause irreversible and irreparable damage, it is essential to determine the immunological and physiological status of the patient. This is the main reason why many therapeutic drugs have failed. The same drug given at different stages of sepsis may be therapeutic or otherwise harmful or have no effect 2,3. To understand sepsis at various levels it is crucial to have a suitable and comprehensive animal model that reproduces the clinical course of the disease. It is important to characterize the pathophysiological mechanisms occurring during sepsis and control the model conditions for testing potential therapeutic agents. 
To study the etiology of human sepsis researchers have developed different animal models. The most widely used clinical model is cecal ligation and puncture (CLP). The CLP model consists of the perforation of the cecum allowing the release of fecal material into the peritoneal cavity to generate an exacerbated immune response induced by polymicrobial infection. This model fulfills the human condition that is clinically relevant. As in humans, mice that undergo CLP with fluid resuscitation show the first (early) hyperdynamic phase that in time progresses to the second (late) hypodynamic phase. In addition, the cytokine profile is similar to that seen in human sepsis where there is increased lymphocyte apoptosis (reviewed in 4,5). Due to the multiple and overlapping mechanisms involved in sepsis, researchers need a suitable sepsis model of controlled severity in order to obtain consistent and reproducible results.

The mouse model of cecal ligation and puncture as a valuable tool for the study of human sepsis.

盲肠结扎穿刺过程

Human sepsis is characterized by a set of systemic reactions in response to intensive and massive infection that failed to be locally contained by the ...

科研

医学

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

部分原位肝移植大鼠模型的肝动脉重建的外科手术

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the failing native liver has regenerated.1-3 The APOLT-method, the orthotopic implantation of auxiliary segments- averts most of the technical problems. However this method necessitates extensive resections of both the native liver and the graft.4 In 1998, Erhard developed the heterotopic auxiliary liver transplantation (HALT) utilizing portal vein arterialization (PVA) (Figure 1). This technique showed promising initial clinical results.5-6 We developed a HALT-technique with flow-regulated PVA in the rat to examine the influence of flow-regulated PVA on graft morphology and function (Figure 2).
A liver graft reduced to 30 % of its original size, was heterotopically implanted in the right renal region of the recipient after explantation of the right kidney.&#160; The infra-hepatic caval vein of the graft was anastomosed with the infrahepatic caval vein of the recipient. The arterialization of the donor&#8217;s portal vein was carried out via the recipient&#8217;s right renal artery with the stent technique. The blood-flow regulation of the arterialized portal vein was achieved with the use of a stent with an internal diameter of 0.3 mm. The celiac trunk of the graft was end-to-side anastomosed with the recipient&#8217;s aorta and the bile duct was implanted into the duodenum. A subtotal resection of the native liver was performed to induce acute hepatic failure. 7
In this manner 112 transplantations were performed. The perioperative survival rate was 90% and the 6-week survival rate was 80%. Six weeks after operation, the native liver regenerated, showing an increase in weight from 2.3&#177;0.8 g to 9.8&#177;1 g. At this time, the graft&#8217;s weight decreased from 3.3&#177;0.8 g to 2.3&#177;0.8 g.
We were able to obtain promising long-term results in terms of graft morphology and function. HALT with flow-regulated PVA reliably bridges acute hepatic failure until the native liver regenerates.

Auxiliary liver transplantation provides a temporary support in acute hepatic failure, until regeneration of the failing liver. The heterotopic auxiliary liver transplantation (HALT) with portal vein arterialization (PVA) renders sufficient liver function. We developed an analogous technique in the rat, to examine the influence of the portal vein arterialization on the morphology and function of the graft.

在急性肝衰竭异位辅助性大鼠肝移植流量调节门静脉动脉

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

Application of the En Bloc Concept Combined with Anatomic Resection in Laparoscopic Hepatectomy

Laparoscopic hepatectomy has been reported in many studies, and it is the mainstream method of liver resection. In some particular cases, such as when there are tumors adjacent to the cystic bed, surgeons cannot palpate the surgical margins through the laparoscopic approach, which leads to uncertainty about R0 resection. Conventionally, the gallbladder is resected first, and the hepatic lobes or segments are resected second. However, tumor tissues can be disseminated in the above cases. To address this issue, based on the recognition of the porta hepatis and intrahepatic anatomy, we propose a unique approach to hepatectomy combined with gallbladder resection by en bloc anatomic resection in situ. Firstly, after dissecting the cystic duct, without cutting the gallbladder primarily, the porta hepatis is pre-occluded by the single lumen ureter; secondly, the left hepatic pedicle is made free by the gap of the Laennec membrane and Hilar plate; thirdly, the assistant is asked to drag the fundus of the gallbladder, and the liver parenchyma tissue is resected using a harmonic scalpel along the ischemia line on the liver surface and intraoperative ultrasound. The whole middle hepatic vein (MHV) and its tributaries appear completely; lastly, the left hepatic vein (LHV) is disconnected, and the specimen is taken out from the abdominal cavity. The tumor, gallbladder, and other surrounding tissues are resected en bloc, which meets the tumor-free criterion, and a wide incisal margin and R0 resection are achieved. Therefore, the laparoscopic hepatectomy with the combination of the en bloc concept and anatomic resection is a safe, effective, and radical method with low postoperative recurrence and metastasis.

Numerous studies have demonstrated the advantages of anatomic resection. Nonetheless, whether anatomic resection can increase R0 resection rates remains controversial. Consequently, the present study describes an innovative procedure involving the en bloc concept combined with anatomic resection in laparoscopic hepatectomy, which can reduce postoperative recurrence and metastasis.

整体概念联合解剖切除在腹腔镜肝切除术中的应用

Laparoscopic hepatectomy has been reported in many studies, and it is the mainstream method of liver resection. In some particular cases, such as when ...

癌症研究

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. To better understand the mechanism behind these surgical procedures, a rat partial liver transplantation (PLTx) model was established for relevant surgical studies. Because of the complexity of the rat PLTx model, a protocol with detailed descriptions is required. An article published previously reported a protocol in which ex vivo hepatectomy was used to achieve 50% rat PLTx. In contrast to this protocol, we introduced a re-arterialized PLTx with an in vivo 70% hepatectomy. An updated vessel-oriented hepatectomy was incorporated into the rat PLTx to refine the microsurgical procedure. The portal veins and hepatic arteries of the left lateral lobe and the median lobe were individually dissected and ligated before removal of the liver parenchyma, thereby decreasing the probability of bleeding in the remnant liver stump. Furthermore, an end-to-side vessel anastomosis between the common hepatic artery and the enlarged proper hepatic artery was introduced to re-arterialize the hepatic artery. By using this end-to-side vessel anastomosis technique, the diameter of the anastomosis was enlarged, thereby decreasing the difficulty of hand suture and maintaining a high rate of anastomotic patency. Moreover, the cuff anastomosis of the infrahepatic vena cava was slightly modified. A section of circumferential liver parenchyma around the vena cava of a recipient was preserved during cuff anastomosis to maintain the three-dimensional shape of the vascular lumen. This section of liver parenchyma was removed after completing the anastomosis. With this modification, the step involving placement of stay sutures was omitted, thereby further shortening the cuff anastomosis time. By using this protocol of rat PLTx, a low liver enzyme level, an intact liver lobular architecture and a high survival rate were achieved after microsurgery.

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Here, a protocol involving re-arterialized rat partial liver transplantation is presented. Specifically, 70% liver was resected in vivo by using an updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner. The cuff technique was modified to shorten the anastomosis time of the infrahepatic vena cava.

静脉大鼠部分肝移植的体内血管定向70% 肝切除术

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

矫形肝移植大鼠模型中动脉重联后并发症减少

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

<meta charset="utf8"></head><body>在分期肝切除术动物模型中研究 ALLPS 的肝脏再生

A Mouse Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy Procedure Aided by Microscopy

Hepatectomy is widely regarded as the primary treatment for hepatic malignancies; yet, postoperative liver failure remains a major cause of perioperative mortality, severely impacting patient outcomes. In a robust hepatic environment, the future liver remnant (FLR) must exceed 25%, and in cases of cirrhosis, this requirement increases to over 40%. The inadequacy of FLR is currently a major obstacle in the progression of hepatic surgery.
Traditional methods to enhance FLR hypertrophy mainly focus on portal vein embolization (PVE), but its effectiveness is considerably limited. In recent years, there have been numerous reports on a novel biphasic hepatectomy method involving hepatic partitioning and portal vein ligation, known as associating liver partition and portal vein ligation for staged hepatectomy (ALPPS). ALPPS surpasses PVE in efficiently and considerably inducing FLR hypertrophy. However, the detailed mechanisms driving ALPPS-facilitated hepatic regeneration are not fully understood. Thus, replicating ALPPS in animal models is crucial to thoroughly investigate the molecular mechanisms of hepatic regeneration, offering valuable theoretical and practical insights.

<meta charset="utf8"></head><body>作者聚焦：通过 ALPPS 小鼠模型推进肝脏再生研究

Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) has been shown to result in remarkable liver regeneration after several weeks post first-stage surgery. Animal models that have undergone ALPPS have been developed to explore potential liver regeneration ability and achieve favorable clinical outcomes.

显微镜辅助分期肝切除术的关联肝分区和门静脉结扎的小鼠模型

Hepatectomy is widely regarded as the primary treatment for hepatic malignancies; yet, postoperative liver failure remains a major cause of perioperative ...

<meta charset="utf8"></head><body>大鼠肝移植和同种异体移植物通畅性分析

Heterotopic Auxiliary Whole Liver Rat Transplant Model Utilizing a Hepaticoureterostomy for Allograft Rejection Studies

Small animal transplant models are indispensable for organ tolerance studies investigating feasible therapeutic interventions in preclinical studies. Rat liver transplantation (LTx) protocols typically use an orthotopic model where the recipients&#39; native liver is removed and replaced with a donor liver. This technically demanding surgical procedure requires advanced micro-surgical skills and is further complicated by lengthy anhepatic and lower body ischemia times. This prompted the development of a less complicated heterotopic method that can be performed faster with no anhepatic or lower body ischemia time, reducing post-surgery stress for the recipient animal.
This heterotopic LTx protocol includes two main steps: excising the liver from the donor rat and transplanting the whole liver into the recipient rat. During the excision of the donor liver, the surgeon ligates the supra-hepatic vena cava (SHVC) and hepatic artery (HA). On the recipient side, the surgeon removes the left kidney and positions the donor liver with the portal vein (PV), infra-hepatic vena cava (IHVC), and bile duct facing the renal vessels. Further, the surgeon anastomoses the recipient&#39;s renal vein end to end with the IHVC of the liver and arterializes the PV with the renal artery using a stent. A hepaticoureterostomy is utilized for biliary drainage by anastomosing the bile duct to the recipient&#39;s ureter, permitting the discharge of bile via the bladder.
The average duration of the transplantation was 130 min, cold ischemia duration was around 35 min, and warm ischemia duration was less than 25 min. Hematoxylin and eosin histology of the auxiliary liver from syngeneic transplants showed normal hepatocyte structure with no significant parenchymal alterations 30 days post-transplant. In contrast, 8-day post-transplant allogeneic graft specimens demonstrated extensive lymphocytic infiltration with a Banff Schema rejection activity index score of 9. Therefore, this LTx method facilitates a low morbidity rejection model alternative to orthotopic LTx.

<meta charset="utf8"></head><body>利用肝输尿管造口术进行同种异体移植物排斥反应研究的异位辅助全肝大鼠移植模型

The rat heterotopic auxiliary liver transplant protocol described here offers a practical investigational tool for exploring mechanisms of hepatic allograft rejection. This model helps to alleviate the surgical hurdles and animal stress of orthotopic liver transplantation in rats.

利用肝输尿管造口术进行同种异体移植物排斥反应研究的异位辅助全肝大鼠移植模型

Small animal transplant models are indispensable for organ tolerance studies investigating feasible therapeutic interventions in preclinical studies. Rat ...

肝分型与门静脉结扎大鼠分期肝切除 (ALPPS) 手术模型的建立

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此视频中的章节

盲肠结扎穿刺过程

部分原位肝移植大鼠模型的肝动脉重建的外科手术

在急性肝衰竭异位辅助性大鼠肝移植流量调节门静脉动脉

整体概念联合解剖切除在腹腔镜肝切除术中的应用

静脉大鼠部分肝移植的体内血管定向70% 肝切除术

矫形肝移植大鼠模型中动脉重联后并发症减少

显微镜辅助分期肝切除术的关联肝分区和门静脉结扎的小鼠模型

显微镜辅助分期肝切除术的关联肝分区和门静脉结扎的小鼠模型

显微镜辅助分期肝切除术的关联肝分区和门静脉结扎的小鼠模型

利用肝输尿管造口术进行同种异体移植物排斥反应研究的异位辅助全肝大鼠移植模型