Name: re-arterialized rat partial liver transplantation with an in vivo vessel-oriented 70% hepatectomy
Uploaded: 2018-04-08T13:00:00
Description: Watch this Scientific Journal Video about Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy at JoVE.com

本研究获得中国国家自然科学基金 (81501382 号赠款) 和浙江省自然科学基金 (批准号:) 的支持。LQ13H100003 和 LQ16H100002)。

所有的程序都遵循了温州医科大学动物政策和福利委员会批准的《啮齿动物外科指南》 15。
1. 动物
<ol><li>用雄性 Lewis 老鼠称250-350 克作为捐献者和受体。</li>
</ol>2. 操作环境
<ol><li>在手术显微镜下执行所有显微外科手术, 放大率从7X 到20X 不等。手术前要消毒所有外科器械。 注意: 操作室和操作表清洁, 但不消毒。</li>
</ol><p class="jove_titl...

<ol>
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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fifty-percent+partial+liver+transplantation+in+the+rat.">Fifty-percent partial liver transplantation in the rat.</a> Transplantation. 62, 292-293 (1996).</li><li>Foss, A., 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=Function+of+reduced-size+liver+transplant+in+the+rat.">Function of reduced-size liver transplant in the rat.</a> Res Exp Med (Berl). 197, 91-99 (1997).</li><li>Nagai, K., Yagi, S., Uemoto, S., Tolba, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+procedures+for+a+rat+model+of+partial+orthotopic+liver+transplantation+with+hepatic+arterial+reconstruction.">Surgical procedures for a rat model of partial orthotopic liver transplantation with hepatic arterial reconstruction.</a> J Vis Exp. , e4376 (2013).</li><li>Sun, J., Qin, G., Wu, L., Wang, C., Sheil, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antigenic+load+and+peripheral+chimeric+levels+in+entire+and+partial+liver+allograft+recipients.">Antigenic load and peripheral chimeric levels in entire and partial liver allograft recipients.</a> Microsurgery. 21, 183-187 (2001).</li><li>Ninomiya, M., 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=Deceleration+of+regenerative+response+improves+the+outcome+of+rat+with+massive+hepatectomy.">Deceleration of regenerative response improves the outcome of rat with massive hepatectomy.</a> Am J Transplant. 10, 1580-1587 (2010).</li><li>Yamanaka, K., 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=Effect+of+olprinone+on+liver+microstructure+in+rat+partial+liver+transplantation.">Effect of olprinone on liver microstructure in rat partial liver transplantation.</a> J Surg Res. 183, 391-396 (2013).</li><li>Wang, W., 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=Mesenchymal+stem+cells+promote+liver+regeneration+and+prolong+survival+in+small-for-size+liver+grafts:+involvement+of+C-Jun+N-terminal+kinase,+cyclin+D1,+and+NF-kappaB.">Mesenchymal stem cells promote liver regeneration and prolong survival in small-for-size liver grafts: involvement of C-Jun N-terminal kinase, cyclin D1, and NF-kappaB.</a> PLoS One. 9, e112532 (2014).</li><li>Madrahimov, N., Dirsch, O., Broelsch, C., Dahmen, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marginal+hepatectomy+in+the+rat:+from+anatomy+to+surgery.">Marginal hepatectomy in the rat: from anatomy to surgery.</a> Ann. Surg. 244, 89-98 (2006).</li><li>Martins, P. N., Theruvath, T. P., Neuhaus, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+of+partial+hepatectomies.">Rodent models of partial hepatectomies.</a> Liver Int. 28, 3-11 (2008).</li><li>Kamada, N., Sumimoto, R., Kaneda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+value+of+hepatic+artery+reconstruction+as+a+technique+in+rat+liver+transplantation.">The value of hepatic artery reconstruction as a technique in rat liver transplantation.</a> Surgery. 111, 195-200 (1992).</li><li>Steger, U., Sawitzki, B., Gassel, A. M., Gassel, H. J., Wood, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+hepatic+rearterialization+on+reperfusion+injury+and+outcome+after+mouse+liver+transplantation.">Impact of hepatic rearterialization on reperfusion injury and outcome after mouse liver transplantation.</a> Transplantation. 76, 327-332 (2003).</li><li>Huang, H., 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=A+novel+end-to-side+anastomosis+technique+for+hepatic+rearterialization+in+rat+orthotopic+liver+transplantation+to+accommodate+size+mismatches+between+vessels.">A novel end-to-side anastomosis technique for hepatic rearterialization in rat orthotopic liver transplantation to accommodate size mismatches between vessels.</a> Eur Surg Res. 47, 53-62 (2011).</li><li>Bernal, J., 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=Guidelines+for+rodent+survival+surgery.">Guidelines for rodent survival surgery.</a> J.Invest Surg. 22, 445-451 (2009).</li><li>Johnson, A. J., Stulting, R. D., Macsai, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ophthalmic Microsurgical Suturing Techniques. , 21-28 (2007).</li><li>Delriviere, L., 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=Detailed+modified+technique+for+safer+harvesting+and+preparation+of+liver+graft+in+the+rat.">Detailed modified technique for safer harvesting and preparation of liver graft in the rat.</a> Microsurgery. 17, 690-696 (1996).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+rats.">Orthotopic liver transplantation in rats.</a> J Vis Exp. , (2012).</li><li>Delriviere, L., 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=Technical+details+for+safer+venous+and+biliary+anastomoses+for+liver+transplantation+in+the+rat.">Technical details for safer venous and biliary anastomoses for liver transplantation in the rat.</a> Microsurgery. 18, 12-18 (1998).</li><li>Jin, H., 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=Preliminary+experience+of+a+PDCA-cycle+and+quality+management+based+training+curriculum+for+rat+liver+transplantation.">Preliminary experience of a PDCA-cycle and quality management based training curriculum for rat liver transplantation.</a> J Surg Res. 176, 409-422 (2012).</li><li>Selzner, N., Selzner, M., Tian, Y., Kadry, Z., Clavien, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cold+ischemia+decreases+liver+regeneration+after+partial+liver+transplantation+in+the+rat:+A+TNF-alpha/IL-6-dependent+mechanism.">Cold ischemia decreases liver regeneration after partial liver transplantation in the rat: A TNF-alpha/IL-6-dependent mechanism.</a> Hepatology. 36, 812-818 (2002).</li><li>Tanaka, H., Hashizume, K., Enosawa, S., Suzuki, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+transplantation+of+a+20%+partial+liver+graft+in+rats:+a+technical+innovation.">Successful transplantation of a 20% partial liver graft in rats: a technical innovation.</a> J Surg Res. 110, 409-412 (2003).</li><li>Bockhorn, M., 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=Erythropoietin+treatment+improves+liver+regeneration+and+survival+in+rat+models+of+extended+liver+resection+and+living+donor+liver+transplantation.">Erythropoietin treatment improves liver regeneration and survival in rat models of extended liver resection and living donor liver transplantation.</a> Transplantation. 86, 1578-1585 (2008).</li><li>Martins, P. N., Neuhaus, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+anatomy+of+the+liver,+hepatic+vasculature+and+bile+ducts+in+the+rat.">Surgical anatomy of the liver, hepatic vasculature and bile ducts in the rat.</a> Liver Int. 27, 384-392 (2007).</li><li>Kamada, N., Calne, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+the+rat.+Technique+using+cuff+for+portal+vein+anastomosis+and+biliary+drainage.">Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage.</a> Transplantation. 28, 47-50 (1979).</li><li>Kashfi, A., 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=A+review+of+various+techniques+of+orthotopic+liver+transplantation+in+the+rat.">A review of various techniques of orthotopic liver transplantation in the rat.</a> Transplant.Proc. 37, 185-188 (2005).</li><li>Wain, R. A. J., Hammond, D., McPhillips, M., Whitty, J. P. M., Ahmed, W., Ahmed, W., Jackson, M. 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大鼠 PLTx 是一种复杂的显微外科手术, 其培训计划成本高且持续时间为20。大鼠 PLTx 协议的复杂性使得研究人员无法使用这种动物模型。与全尺寸大鼠 LTx 相比, 大鼠 PLTx 的 microsurgeon 不仅与移植过程的挑战有关, 而且与小动物肝脏切除的挑战有关。因此, 详细描述整个过程的可视化显微外科协议是建立这一复杂模型的必要条件。Nagai et所报告的先前可视化的文章提出了一项50%...

目前, 捐赠肝脏的数量和等待捐献肝脏的病人数量之间存在很大的差异。肝脏器官的短缺是一个全球性的问题。为了扩大捐献者的水池, 分裂肝移植 (LTx) 和活体捐献者 LTx 被开发用于使用部分肝脏作为移植物1。
为进一步研究部分肝移植 (PLTx) 的机制, 建立了相关动物模型2,3,4,5。在大鼠 PLTx 中, 肝叶切除体内和前体, 以模拟活体捐献者 LTx 的条件, 并分别在人类体内分裂 LTx。在可视化实验杂志上发表的一篇论文提出了一个详细的协议, 涉及50% 只老鼠 PLTx 使用前体肝切除5。然而, 在可视化文献中还没有报道一只老鼠 PLTx 与体内肝切除术有关。
除了体内和前体hepatectomies 之间的差异外, 执行肝切除术本身也在确定 PLTx 的结果方面起着重要作用. 目前, 在许多外科研究中使用大鼠 PTLx模型, 肝叶切除后, 放置一个简单的结扎在肝瓣的椎弓根2,3,6,7,8,9。然而, 在切除前放置一个简单的结扎不适合所有肝叶, 因为不同的肝叶有不同的形状和大小。在正中裂片的基础上进行简单结扎, 有很高的风险导致腔静脉收缩, 这可能最终影响部分肝移植的流出10,11。因此, 在大鼠 PTLx 的研究中, 需要以大鼠肝脏解剖知识为基础的更新肝切除技术。
在本研究中所述的协议中, 将最新的以血管为导向的70% 肝切除术纳入大鼠 PLTx 的程序中。在肝实质切除前分别对左外侧叶 (微光) 和正中叶 (ML) 的门静脉和肝动脉进行解剖和分切。然后用穿刺缝线结扎了微光和 ML 的肝静脉。通过使用单独的结扎和多重穿刺缝合, 而不是简单的结扎, 毫升的残余树桩能够扩散到腔静脉。因此, 避免了手术结扎引起的 infrahepatic 腔静脉的收缩。另外, 在切除肝实质之前, 单独结扎对微光和 ML 血供的阻断减少了残肝残端的出血率, 从而最大限度地减少了血液损失对实验的影响11。
对于 microsurgeons, 由于该血管直径极小, 重建肝移植肝动脉 (PHA) 是一项重大的挑战。虽然在 LTx 中重新动脉是否真的是必要的问题仍在辩论中12,13,14, 已经提出了许多重建肝动脉的显微外科技术14.本文介绍了一种新的肝移植动脉技术, 它以端到端的方式将普通肝动脉 (CHA) 与扩大后的医用血管吻合。使用这种端对端技术, 两个肝动脉连接到一个更大的直径吻合。吻合口直径越大, 缝合越容易, 吻合通畅的改善越大。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Surgical microscope</td>
			<td>M&#246;ller-Wedel, Germany</td>
			<td>654359</td>
			<td></td>
		</tr>
		<tr>
			<td>Light source</td>
			<td>OSRAM (China) Lighting Co., Ltd</td>
			<td>64627</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane Vaporizer</td>
			<td>MIDMARK Corporation</td>
			<td>VIP3000</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Baxter Healthcare Corporation</td>
			<td>40032609</td>
			<td>For Inhalation anesthesia</td>
		</tr>
		<tr>
			<td>Normal Saline</td>
			<td>Zhejiang Tianrui Pharmaceutical Co., Ltd.</td>
			<td>716092103</td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone Iodine Solution</td>
			<td>HANGZHOU MINSHENG PHARMACEUTICAL</td>
			<td>H33021567</td>
			<td>For disinfection</td>
		</tr>
		<tr>
			<td>Hemostatic forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>J31020</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical scissors</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>JC2116</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle-holder</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>J32030</td>
			<td></td>
		</tr>
		<tr>
			<td>Dressing Forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>J42020</td>
			<td></td>
		</tr>
		<tr>
			<td>Mosquito hemostatic forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>W40340</td>
			<td>For liver resection; For retracting the stay sutures.&#160;</td>
		</tr>
		<tr>
			<td>Micro-scissors</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA1040</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro needle-holder</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA2060</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved micro typing forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA3061</td>
			<td></td>
		</tr>
		<tr>
			<td>straight micro typing forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA3060</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro hemostatic forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA3020</td>
			<td></td>
		</tr>
		<tr>
			<td>Bulldog clamp</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>XEC350</td>
			<td></td>
		</tr>
		<tr>
			<td>Vessel clamps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>W40160</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Serrefine-curved</td>
			<td>&#160;Fine Science Tools, Inc</td>
			<td>18055-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Dacryosyrinx</td>
			<td>Shi Zhuo medical instruments Co., Ltd.</td>
			<td>5# curve</td>
			<td>Use as the curved needle for forcing out the air bubbles in the vascular lumen</td>
		</tr>
		<tr>
			<td>Sterilized gauze swabs</td>
			<td>Fuqing Health &#38; Integral Medical</td>
			<td>20230R</td>
			<td></td>
		</tr>
		<tr>
			<td>5mL syringe</td>
			<td>Zhejiang Yusheng Medical Instrument Co.,Ltd</td>
			<td>811932416</td>
			<td></td>
		</tr>
		<tr>
			<td>18G needles</td>
			<td>Shanghai Kindly Enterprise Development Group Co., Ltd</td>
			<td>01-014</td>
			<td>For fixing the upper abdomen wall</td>
		</tr>
		<tr>
			<td>Intima II 22G intraveous catheter</td>
			<td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td>
			<td>383207y</td>
			<td>For making biliary stent; For infusing perfusate</td>
		</tr>
		<tr>
			<td>7-0 polypropylene sutures</td>
			<td>Ningbo Medical Needle Co., Ltd</td>
			<td>151026</td>
			<td>For SHVC anastomsis</td>
		</tr>
		<tr>
			<td>6-0 silk suture</td>
			<td>Shanghai Pudong Jinhuan Medical Products Co., Ltd</td>
			<td>2650182</td>
			<td>For ligation</td>
		</tr>
		<tr>
			<td>Culture dish (100mm)</td>
			<td>Corning Incorporated</td>
			<td>430165</td>
			<td>used in back table for storage ice. The back table dish was placed on it.&#160;</td>
		</tr>
		<tr>
			<td>Culture dish (30 mm)</td>
			<td>Corning Incorporated</td>
			<td>3296</td>
			<td>As a vessel for organ preservation</td>
		</tr>
		<tr>
			<td>Angiocath 12G intravenous catheter</td>
			<td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td>
			<td>382277</td>
			<td>For making a IHVC cuff</td>
		</tr>
		<tr>
			<td>Angiocath 14G intravenous catheter</td>
			<td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td>
			<td>382269</td>
			<td>For making a PV cuff</td>
		</tr>
		<tr>
			<td>Buprenorphine (hydrochloride)</td>
			<td>Tianjin Institute of Pharmaceutical Research Pharmaceutical Co., Ltd</td>
			<td>H12020275</td>
			<td>For analgesic</td>
		</tr>
		<tr>
			<td>Cefuroxime sodium for injection</td>
			<td>Esseti FarmaceuticiS.r.l</td>
			<td>H20160013</td>
			<td>As an antibiotics</td>
		</tr>
	</tbody>
</table>

re-arterialized rat partial liver transplantation with an in vivo vessel-oriented 70% hepatectomy

在临床上发展了肝移植和活体肝供肝移植, 以更有效地利用肝脏器官。为更好地了解这些手术过程的机制, 建立了大鼠部分肝移植 (PLTx) 模型进行相关的外科研究。由于大鼠 PLTx 模型的复杂性, 需要一个具有详细描述的协议。此前发表的一篇文章报告了一项协议, 其中前体肝切除术用于实现50% 大鼠 PLTx。与此协议相比, 我们引入了一个 PLTx体内70% 肝切除的重新静脉。以血管为导向的肝切除术被纳入大鼠 PLTx, 以改进显微外科手术。肝实质切除前, 左外侧叶和正中叶的门静脉和肝动脉分别进行解剖和结扎, 从而降低残肝残端出血的几率。此外, 还介绍了肝动脉与肝动脉扩张的端到端血管吻合术, 以重新 arterialize 肝脏动脉。采用端到端血管吻合技术, 扩大了吻合口的直径, 降低了手缝的难度, 保持了较高的吻合通畅率。此外, infrahepatic 腔静脉的袖套吻合略有改变。在袖状吻合术期间, 在接受者的腔静脉周围有一节环性肝实质, 以维持血管腔的三维形状。这部分肝实质在完成吻合术后被切除。经此修改, 省略了放置缝合线的步骤, 从而进一步缩短了袖套吻合时间。采用本协议对大鼠 PLTx, 在显微外科手术后实现了低肝酶水平、完整的肝小叶结构和高存活率。

共完成31例自体静脉大鼠 PLTx 的应用本协议。所有收件人都存活到观察时间结束为止。患者的体重在术后一天 (POD) 4 开始恢复。在 POD 6 (图 8) 之后, 体重的斜率接近正常的 Lewis 鼠, 这间接地暗示了接收方的恢复。组织学上, 接受者观察到胆管的轻微增生, 接受者的肝小叶结构完好。没有发现坏死或明显的正弦扩张 (图 9)。血清丙氨...

Watch this Scientific Journal Video about Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy at JoVE.com

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

本文提出了一种静脉大鼠部分肝移植的协议。具体地说, 70% 例肝切除术是通过使用一种最新的血管性肝切除技术在体内进行的。肝动脉以端对侧的方式重建。对袖套技术进行了改进, 缩短了 infrahepatic 腔静脉的吻合时间。

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. ...

A protocol involving re-arterialized rat partial liver transplantation is presented in this article. Specifically, 70%of liver was resected in vivo by using the 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 inferior patic vena cava. Isoflurane mixed with zero point five liter per minute oxygen was used to induce and maintain inhalation anesthesia. Cut a small transverse incision on the upper abdomen.Then, extend the transverse incision close to the costal margin on the upper abdomen. Reverse and affix upper abdomen wall to the cranial side. Next, reverse the xiphoid process to cranial side, too.Cut off the falciform ligament, coronary ligament, and a triangular ligament, in sequence. Free the left diaphragmatic vein from the diaphragm using microforceps. Then, double ligate it using six-zero silk sutures.Free the hepatoesophageal ligament and artery. Then, double-ligate it and divide it. Expose the first hepatic hilum of the liver.First, carefully dissect the posterior wall of the common trunk of the portal vein. Then, double ligate it and divide it with the corresponding bile duct and the hepatic artery together. Second, gently dissect, double-ligate and divide the portal vein of the right medium lobe.Third, double ligate and divide the hepatic artery and the bile duct of the right medium lobe using six-zero silk sutures. Ligate the hepatic of the left lateral lobe using circumstantial three-zero silk suture. Remove the liver mass using micro scissors above the ligation.Clamp the left median hepatic vein by mosquito hemostatic forceps placed along the median fissure. Remove all of the liver parenchyma above the forceps using micro scissors. Place the piercing suture under the forceps surrounding the left median hepatic vein.Then, tie a square knot to ligate it. Besides, place another two piercing sutures to close the rest of the incision. After this, place a mosquito hemostatic forceps at a distance of point two-five centimeters to occlude the right median hepatic vein.Remove all liver parenchyma above the forceps. Place the first piercing suture penetrating the liver parenchyma under the forceps to surround the middle median hepatic vein. Then, tie a square knot to ligate it.Place a second piercing suture under the forceps to surround the middle median hepatic vein. Then, ligate it using the same method. Place another two piercing sutures under the mosquito hemostatic forceps to close the rest of the incision of the medium lobe by using these piercing sutures.The remnant stump of the medium lobe was able to spread over the vena cava. Cut open the retroperitoneum to expose the inferior hepatic vena cava. Dissect and ligate the right adrenal vein, which drains into the inferior hepatic vena cava from the rear.Free the right renal artery and the right renal vein. Perform a right nephrectomy after ligating the right renal vein and the right renal artery. Mobilize the common bile duct from the first hepatic hilum using micro forceps.Then, ligate the common bile duct at the crest point of the polaric vein and the common bile duct. Cut a transverse incision on the interior wall of the common bile duct. Insert a 22-gauge biliary stent immediately into lumen of the common bile duct.Secure the biliary stent with circumferential six-zero silk suture. Transect the common bile duct between the two six-zero silk sutures. Free the pyloric vein using micro forceps then double ligate and divide it.Next, double ligate and divide the splenic vein the same way. Carefully free the common hepatic artery and it's two bifurcations. Double ligate the gastroduodenal artery using six-zero silk sutures at the bifurcation of the common hepatic artery to gastroduodenal artery.Inject 50 IU of heparin in one milliliter of saline via the penile vein. Cross-clamp the inferior hepatic vena cava portal vein and the common hepatic artery using three curved micro Perform a small incision on the interior wall of the portal vein to insert a 22-gauge catheter. Profuse the partial liver graft with saline at four degrees celsius via the catheter under pressure of 20 centimeters water.Rapidly cut open the thoracic cavity and intrathoracic vena cava using surgical scissors to drain out the blood and profuse it Use micro scissors to transect the left diaphragmatic vein between the two ligations which were placed previously. Next, transect the suprahepatic vena cava close to the diaphragm to keep the vascular lanes as long as possible. Cut off the portal vein at the level of the splenic vein and transect the common hepatic artery close to the select trunk.Cut off the adrenal vessels on the distill side of the ligation. Then, divide the infrahepatic vena cava at the level of the left renal vein. Pull the portal vein through the lumen of the cuff made with a 12-gauge intravenous catheter.Temporarily fix puff handle and portal vein together. Avert the portal vessel wall to cover the outside surface of the cuff. Secure the vessel wall and the cuff in position with circumferential six-zero silk suture.Install the infrahepatic vena cava cuff using the same method. Penetrate the vessel wall of the suprahepatic vena cava from outside to inside using two seven-zero polypropylene sutures along the three o'clock and nine o'clock directions, respectively. Repeat the procedures in 6.1, 6.2, and 6.7 in the recipient, except for the right nephrectomy.Mobilize, double ligate, and divide the common bile duct at the first bifurcation in the hepatic hilum. Transect and prepare the hepatic artery using micro scissors, subsequent to double ligation using six-zero silk suture. Retract the recipient liver to the left side using two cotton sticks.Cut off the ligament in the rear of the liver from the suprahepatic vena cava to the right adrenal vein. Introduce a rubber band through the back space of the suprahepatic vena cava. Cross-clamp the infrahepatic vena cava above the renal vein using a vessel clamp and the portal vein above the pyloric vein.the suprahepatic vena cava and diaphragm and clamp the parts of the diaphragm together with the intrathoracic vena cava. Cut the vessel wall of the suprahepatic vena cava carefully along the upper edge of the liver parenchyma to keep the vascular wall as long as possible. Transect the portal vein at the first bifurcation in the hepatic hilum using micro scissors.Transect the inferior vena cava and parenchyma of the right inferior lobe rather than in the vessel wall to leave a section of circumferential liver parenchyma around the vena cava. Keep the cutting edge in the right inferior lobe at a distance of four millimeters to the inferior hepatic vena cava. Place the partial liver graft in the abdominal cavity, isotopically.Pierce the vessel wall of the suprahepatic vena cava from inside to outside along three o'clock and nine o'clock directions Using two prestat stay sutures. Pull the stay sutures to the right and left after tying into a knot with their own ends. Pierce the suprahepatic vena cava vessel wall from outside to inside, close to the knot in the three o'clock direction.Anastomose the posterior wall of the suprahepatic vena cava by running suture inside the vascular lumen from the three o'clock direction to the nine o'clock direction. At the corner of the nine o'clock direction, pierce the vessel wall from inside to outside, close to the knot, to introduce the needle out of the vascular lumen. Use this suture to anastomose interior wall using running suture outside the vascular lumen from the nine o'clock direction to the three direction.Finally, tie the suture with its own end three times. Place two seven-zero polypropylene stay sutures at the end of the portal vein of the recipient, following the description in seven point two. Pull two stay sutures in the opposite direction to extend the vessel wall.Inject approximately one milliliter of saline into the vascular lumen of the portal vein to force out air bubbles. Quickly insert the portal vein cuff into the vascular lumen of the portal vein in the recipient when it is opened using microforceps. Secure the cuff anastomosis using circumferential six-zero silk suture.Then, release the clamps on the suprahepatic vena cava and the portal vein, in sequence. Inject one milliliter of saline into the vascular lumen to force out air bubbles. Insert the cuff into cylindrical vascular lumen of the inferior hepatic vena cava.Quickly secure the cuff anastomosis using a six-zero silk suture. Release the two vessel clamps in the inferior hepatic vena cava to restart the profusion. Then, trim off the surrounding liver parenchyma carefully, above the circumferential silk suture using micro scissors.Penetrate the vessel wall of the common hepatic artery in the liver graft from outside to inside using two eleven-zero polypropylene sutures in the three o'clock and nine o'clock directions, respectively. Occlude the blood flow of the common hepatic artery and the gastroduodenal artery in the recipient using two curved micro Transect the proper hepatic artery of the recipient at its root to expose the vascular lumen. Enlarge the vascular lumen of the proper hepatic artery in the recipient by cutting part of the vessel wall, longitudinally, to accommodate the diameter of the common hepatic artery in the liver graft.Flush the vascular lumen using saline. Anastomose the common hepatic artery of the liver graft to enlarge the proper hepatic artery in an end-to-side manner using the running suture technique. Perform a transverse incision on the anterior wall of the bile duct in the recipient.Pull the biliary stent in the partial liver graft down to insert it into the lumen of the bile duct in the recipient. Divide the transverse incision. Secure the stent using six-zero circumferential silk suture.Tie the two six-zero circumferential silk sutures in the bile duct to each other to reduce the tension of the anastomosis. After surgery, treat all recipients with 0.1 milligram per kilogram Buprenorphine and Cefuroxime 16 milligram per kilogram, subcutaneously. In total, 31 cases have seen generic arterialized rat partial liver transplantation were completed using this protocol.All of the recipients survived until the end of the observation time. The body weight of the recipient began to recover after postoperative day four. The slope of the body weight was close to that of the Lewis rat after postoperative day six, which indirectly implied recovery of the recipient.Histologically, a slight proliferation of the bile duct was observed in the recipients and the liver lobular architecture of the recipient was intact. No necrosis or obvious sign of pseudo dilation was observed. The serum level of alanine aminotransferase was in the normal range at postoperative day 30.No cases of yellow urine caused by jaundice were observed in these recipients, which was indicated by checking the cage bedding daily, indicating the patency of the biliary stent. The normal serum level of bilirubin on postoperative day 30 further confirmed the success for reconstruction of the bile duct. The average duration of the anhepatic phase was 22.03 plus or minus 2.3 minutes.The cuff anastomosis of the portal vein took 4.68 plus or minus 0.77 minutes, while the cuff anastomosis of the inferior hepatic vena cava only took 2.05 plus or minus 0.71 mminutes. Comparatively, the cuff anastomosis of the portal vein, the cuff anastomosis of the inferior hepatic vena cava saved the staff of placing stay sutures, and thereby, further shortens the anastomosis time. Two cases of bleeding in the remnant stump of the medium lobe during the 70%hepatectomy were observed, among 31 cases of vessel-oriented hepatectomy.In the previous experiment, bleeding in the remnant stump of the medium lobe was observed in eight out of 18 cases of parenchyma, preserving vessel-oriented hepatectomy. This result indicated a lower rate of bleeding in the remnant liver stump of the medium lobe, after using vessel-oriented 70%hepatectomy. By using this protocol of rat partial liver transplantation, a low liver enzyme level and intact liver lobular architecture and a high survival rate were achieved after microsurgery.

Research

JoVE Journal

Medicine

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

Orthotopic Liver Transplantation in Rats

大鼠原位肝移植

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

科研

医学

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

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

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 ...

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

之后，正常小鼠70％部分肝切除血管和实质再生的可视化

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...

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

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

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 ...

Intrasplenic Transplantation of Hepatocytes After Partial Hepatectomy in NOD.SCID Mice

肝细胞部分肝切除术后脾移植。小鼠

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

脂肪组织干细胞移植对大鼠部分结肠切除术后渗漏的减少

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

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

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 ...

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

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 ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

毛利精密切肝片作为肝脏生物学的Ex Vivo模型

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...