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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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimal+use+of+hepatobiliary+scintigraphy+before+liver+resection.">Optimal use of hepatobiliary scintigraphy before liver resection.</a> HPB (Oxford). 18 (10), 870 (2016).</li><li>Lau, L., Christophi, C., Muralidharan, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraoperative+functional+liver+remnant+assessment+with+indocyanine+green+clearance:+another+toehold+for+climbing+the+&amp;#34;ALPPS&amp;#34;.">Intraoperative functional liver remnant assessment with indocyanine green clearance: another toehold for climbing the &amp;#34;ALPPS&amp;#34;.</a> Ann Surg. 261 (2), e43-e45 (2015).</li><li>Cieslak, K. 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?...

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 border="0" cellpadding="0" cellspacing="0" width="1220">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:225px;">Isoflurane, 250ml bottles</td>
			<td style="width:400px;">Attane, Piramal, Mumbai, India</td>
			<td style="width:255px;">LDNI 22098</td>
			<td style="width:340px;">Standard vet. equipment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tec-3 Isofluorane Vaporizer</td>
			<td>Ohmeda, GE-Healthcare, Chicago, IL</td>
			<td>not available anymore</td>
			<td>Standard vet. equipment&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenorphine (Temgesic)</td>
			<td>Indivior, Baar, Switzerland</td>
			<td>7680419310353</td>
			<td>GTIN-number</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vitamine A ointment</td>
			<td>Bausch&#38;Lomp, Zug, Switzerland</td>
			<td>7680223980247</td>
			<td>GTIN-number</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Atropine sulfate 0.5mg/ml</td>
			<td>Sintetica SA, Mendrisio, Switzerland</td>
			<td>7680565330045</td>
			<td>GTIN-number</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microsurgery microscope</td>
			<td>Olympus, Tokio, Japan</td>
			<td>SZX10</td>
			<td>Standard vet. equipment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Betadine</td>
			<td>Mundipharma, Basel, Switzerland</td>
			<td>7680342821377</td>
			<td>GTIN-number</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sponges</td>
			<td>Carl Roth GmbH, Karlsruhe, Germany</td>
			<td>NK83.1</td>
			<td>Mini-sponges</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Abdominal Wall retractors</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Self-made from paper clips and Q-Tips</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3-0 silk&#160;</td>
			<td>Ethicon, Sommerville, NJ</td>
			<td>K872H</td>
			<td>Standard surgical</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scissors&#160;</td>
			<td>World precision instruments (WPI), Sarasota, FL</td>
			<td>503371</td>
			<td>Standard microsurgical</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Adson forceps</td>
			<td>World precision instruments (WPI), Sarasota, FL</td>
			<td>501244-G</td>
			<td>Standard microsurgical</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">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 height="21">
			<td height="21" style="height:21px;">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&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">6-0 LOOK black braided silk</td>
			<td>Surgical Specalities Corporation, Wyomissing, PA&#160;</td>
			<td>SP114</td>
			<td>Spool, precut prior to the procedure</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-0 silk sutures</td>
			<td>Ethicon, Sommerville, NJ</td>
			<td>K833</td>
			<td>Standard surgical</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5-0 maxon sutures</td>
			<td>Covidien, Dublin, Ireland</td>
			<td>6608-21</td>
			<td>Standard surgical</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bipolar microforceps</td>
			<td>Sutter, Freiburg, Germany</td>
			<td>780148SGS</td>
			<td>Essential for parenchymal transection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Q-tips small</td>
			<td>Carl Roth GmbH, Karlsruhe, Germany</td>
			<td>EH11.1</td>
			<td>Standard surgical</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Q-tips big</td>
			<td>Carl Roth GmbH, Karlsruhe, Germany</td>
			<td>XL54.1</td>
			<td>Standard surgical</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">G30 needle&#160;</td>
			<td>Terumo, Tokyo, Japan</td>
			<td>NN-3013R&#160;</td>
			<td>Standard anesthesia equipment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2mm volume flow probe&#160;</td>
			<td>Transonic Systems, Ithaca, NY</td>
			<td>MA-2PS</td>
			<td>Smallest available probe for HAT-311 flow meter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Transonic flow meter</td>
			<td>Transonic Systems, Ithaca, NY</td>
			<td>HAT-311 Transsonic flow QC meter</td>
			<td>One of the&#160; first generation flow flow meters for surgery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ExiTron nano 12,000&#160;</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 height="21">
			<td height="21" style="height:21px;">G26 intravenous catheter</td>
			<td>Becton Dickinson, Franklin Lakes, NJ</td>
			<td>391349</td>
			<td>Standard anesthesia equipment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Quantum FX MicroCT&#160;</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&#252;rich</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">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?...

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

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

3-Dimensional Resin Casting and Imaging of Mouse Portal Vein or Intrahepatic Bile Duct System

In organs, the correct architecture of vascular and ductal structures is indispensable for proper physiological function, and the formation and maintenance of these structures is a highly regulated process. The analysis of these complex, 3-dimensional structures has greatly depended on either 2-dimensional examination in section or on dye injection studies. These techniques, however, are not able to provide a complete and quantifiable representation of the ductal or vascular structures they are intended to elucidate. Alternatively, the nature of 3-dimensional plastic resin casts generates a permanent snapshot of the system and is a novel and widely useful technique for visualizing and quantifying 3-dimensional structures and networks.
A crucial advantage of the resin casting system is the ability to determine the intact and connected, or communicating, structure of a blood vessel or duct. The structure of vascular and ductal networks are crucial for organ function, and this technique has the potential to aid study of vascular and ductal networks in several ways. Resin casting may be used to analyze normal morphology and functional architecture of a luminal structure, identify developmental morphogenetic changes, and uncover morphological differences in tissue architecture between normal and disease states. Previous work has utilized resin casting to study, for example, architectural and functional defects within the mouse intrahepatic bile duct system that were not reflected in 2-dimensional analysis of the structure1,2, alterations in brain vasculature of a Alzheimer's disease mouse model3, portal vein abnormalities in portal hypertensive and cirrhotic mice4, developmental steps in rat lymphatic maturation between immature and adult lungs5, immediate microvascular changes in the rat liver, pancreas, and kidney in response in to chemical injury6.
Here we present a method of generating a 3-dimensional resin cast of a mouse vascular or ductal network, focusing specifically on the portal vein and intrahepatic bile duct. These casts can be visualized by clearing or macerating the tissue and can then be analyzed. This technique can be applied to virtually any vascular or ductal system and would be directly applicable to any study inquiring into the development, function, maintenance, or injury of a 3-dimensional ductal or vascular structure.

A method of visualizing and quantifying the 3-dimensional structure of mouse hepatic portal vein or intrahepatic bile duct is described. This resin cast technique can also be applied to other ductal or vascular systems and allows for in situ visualization or quantification of a system's intact communicating architecture.

树脂浇铸和3维成像的小鼠门静脉或肝内胆管系统

In  organs, the correct architecture of vascular and ductal structures is  indispensable for proper physiological function, and the formation and  ...

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 ...

The Dimethylnitrosamine Induced Liver Fibrosis Model in the Rat

Four to six week old, male Wistar rats were used to produce animal models of liver fibrosis. The process requires four weeks of administration of 10 mg/kg dimethylnitrosamine (DMN), given intraperitoneally for three consecutive days per week. Intraperitoneal injections were performed in the fume hood as DMN is a known hepatoxin and carcinogen. The model has several advantages. Firstly, liver changes can be studied sequentially or at particular stages of interest. Secondly, the stage of liver disease can be monitored by measurement of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) enzymes. Thirdly, the severity of liver damage at different stages can be confirmed by sacrifice of animals at designated time points, followed by histological examination of Masson&#39;s Trichome stained liver tissues. After four weeks of DMN dosing, the typical fibrosis score is 5 to 6 on the Ishak scale. The model can be reproduced consistently and has been widely used to assess the efficacy of potential anti-fibrotic agents.

We describe a method to produce an animal model of liver fibrosis in the rat, and assess the degree of fibrosis by histological examination of the liver. The model can be used to study the development of liver disease as well as to test the efficacy of potential anti-fibrotic agents.

该二甲基亚硝胺诱发肝纤维化模型大鼠

Four to six week old, male Wistar rats were used to produce animal models of liver fibrosis. The process requires four weeks of administration of 10 mg/kg ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

直接节段性肝内交付使用大鼠肝门部夹具模型方法

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

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. ...

Generation of a Rat Model of Acute Liver Failure by Combining 70% Partial Hepatectomy and Acetaminophen

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and detoxifying functions of the liver. In most irreversible liver damage cases, orthotropic liver transplant (OLT) remains the only available treatment. To study the therapeutic potential of a treatment for ALF, its prior testing in an animal model of ALF is essential. In the current study, an ALF model in rats was developed by combining 70% partial hepatectomy (PHx) and injections of acetaminophen (APAP) that provides a therapeutic window of 48 h. The median and left lateral lobes of the liver were removed to excise 70% of the liver mass and APAP was given 24 h postsurgically for 2 days. Survival in ALF-induced animals was found to be severely decreased. The development of ALF was confirmed by altered serum levels of the enzymes alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP); changes in prothrombin time (PT); and assessment of the international normalized ratio (INR). Study of the gene expression profile by qPCR revealed an increase in expression levels of genes involved in apoptosis, inflammation, and in the progression of liver injury. Diffused degeneration of hepatocytes and infiltration of immune cells was observed by histological evaluation. The reversibility of ALF was confirmed by the restoration of survival and serum levels of ALT, AST, and ALP after intrasplenic transplantation of syngeneic healthy rat hepatocytes. This model presents a reliable alternative to the available ALF animal models to study the pathophysiology of ALF as well as to evaluate the potential of a novel therapy for ALF. The use of two different approaches also makes it possible to study the combined effect of physical and drug-induced liver injury. The reproducibility and feasibility of current procedure is an added benefit of the model.

The acute liver failure animal model developed in the current study presents a feasible alternative for the study of potential therapies. The current model employs the combined effect of physical and drug-induced hepatic injury and provides a suitable time window to study the potential of novel therapies.

通过结合70%部分切除术和对乙酰氨基酚，生成急性肝衰竭大鼠模型

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

器官缺血-输液损伤模拟大鼠肝脏移植模型的血动力学变化

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...

Repetitive Blood Sampling from the Subclavian Vein of Conscious Rat

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to detect drug exposure. The rat blood sampling method determines the quality of the plasma and further affects the precision of the test results. The subclavian vein blood collection method described in this protocol collects blood samples repeatedly in the conscious state of animals to meet the needs of PK and TK tests. The skills of restraint handling and appropriate procedure of needle incision ensure the success rate of blood collection. It is easy to operate while ensuring the quality of plasma and at the same time catering to animal welfare. However, this method requires skilled operation, and an improper one may cause animal weakness, pain, lameness, and even mortality. The current method has been used in the testing facility for a 4-week oral toxicity study in Sprague Dawley (SD) rats with TK. The maximum amount of blood collected within 24 h did not exceed 20% of the animal's total blood. The animals' body weight was more than 200 g for males and females. The data showed that the animals' bodyweight increased steadily every week, and the clinical observation was normal after repetitive sample collection.

The present protocol describes a simple and efficient method for collecting blood from the subclavian vein in rats. It enables rapid, timely, and easily identifiable sampling without anesthesia and obtains high-quality blood through repetitive sample collection.

意识大鼠锁骨下静脉的重复血液采样

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to ...

Robotic Taj Mahal Hepatectomy for Hilar Cholangiocarcinoma

Hilar cholangiocarcinoma is the most common malignant tumor of the biliary tract. Radical surgical resection is the only effective treatment option. In this study, a 32-year-old male patient with Bismuth Type IVa hilar cholangiocarcinoma underwent radical robotic resection of hepatic S4b, S5, and S1 (Taj Mahal hepatectomy) combined with regional lymphadenectomy, hilar bile duct reconstruction, and hepaticojejunostomy by the robotic surgical system. Postoperative pathological examination showed moderately-differentiated adenocarcinoma of the hilar bile duct. The surgical margins of the liver and bile ducts were negative. Recovery was smooth and the patient was discharged on the 17th postoperative day. The robotic surgical system and associated multiple instruments along with flexible and precise movements is suitable for the local hepatectomy around the porta hepatis, and delicate reconstruction of the hilar bile duct with a smaller diameter. This first clinical application study found that robotic Taj Mahal hepatectomy for hilar cholangiocarcinoma is safe and feasible and needs more experience for the evaluation of its long-term outcomes.

Robotic resection of hepatic S4b, S5, and S1 using the Taj Mahal procedure is feasible and safe for selected patients with hilar cholangiocarcinoma. The step-by-step details for this surgery are presented here.

机器人泰姬陵肝切除术治疗肺门胆管癌

Hilar cholangiocarcinoma is the most common malignant tumor of the biliary tract. Radical surgical resection is the only effective treatment option. In ...