大鼠原位肝移植模型的建立及术式改进

目的探讨用双袖套法建立大鼠原位肝移植模型的手术改进方法. 方法在Kamada等的袖套法吻合血管的基础上进行改进,供体改经腹主动脉进行肝脏冷灌注,肝上下腔静脉用缝合法吻合,门静脉和肝下下腔静脉用袖套法吻合,胆总管采用单管内支架胆管端端吻合下进行原位肝移植360例(次). 结果施行大鼠原位肝移植其供体手术时间(31.2±5.0)分钟,供肝修整时间(12.0±3.0)分钟,受体手术时间(45.0±5.5)分钟,无肝期(20.0±2.5)分钟,手术死亡31例,其中出血12例,肝下下腔静脉血栓8例,肝上下腔静脉回流不畅7例,袖套扭转或脱落4例.术后成活2天以上329例,成活率为91.4%.非干预组(非药物治疗组)1周存活率达86.5%. 结论改进的大鼠原位肝移植术操作简便,手术成功率高,可作为肝移植实验可靠、稳定的动物模型.     

提高二袖套法大鼠原位肝移植成功率的手术技巧

目的　探讨二袖套法大鼠原位肝移植的手术技巧。方法　在Kamada“二袖套法”基础上进行改良。获取供肝前阻断肝门血供 10min ,再灌注 10min ;分别经腹主动脉和门静脉对肝脏进行双重灌注 ;门静脉和肝下下腔静脉用袖套法吻合 ,肝上下腔静脉用缝合法吻合 ,胆总管采用单管内支架胆管端端吻合法。结果　共施行大鼠原位肝移植 12 0次 ,手术成功率为 90 .8%。平均无肝期为 ( 2 1.0± 3.5 )min ,受体总手术时间为 ( 4 6 .0± 4 .5 )min ,1周生存率为 87.2 %。结论　良好的手术野暴露 ,娴熟的显微外科技术 ,精细的手术操作和配合有助于缩短受体无肝期及总手术时间和提高受体生存率     

大鼠改良式原位肝移植手术技巧探讨

目的 探讨建立稳定大鼠原位肝移植模型的手术操作技巧. 方法成年雄性SD大鼠200只,体重200～250 g;成年雄性Wistar大鼠60只,体重230～280 g,供体体重小于受体约30 g.其中SD大鼠为供、受体的同基因肝移植70只(SD-SD组),SD、Wistar分别为供、受体的同种异体肝移植60只(SD.Wistar组).采用改良二袖套法行大鼠原位肝移植,充分暴露第一肝门,不翻动肝脏先行门静脉灌注;在体一步法离断肝上下腔静脉,不带膈肌环;吻合肝上下腔静脉采用单线连续缝合;双线牵引法安装门静脉袖套.术后充分补液维持大鼠血液动力学稳定. 结果 供体手术时间(38.2 ±2.5)min,受体手术时间(45.6±3.5)min,无肝期(15.1±2.2)min,手术成功率93%,1周存活率92%,与传统二袖套法比较差异有统计学意义(P＜0.05).SD-SD组手术成功64只,受体存活时间2～9个月,平均145 d;术后约3 d 肝功能恢复正常,肝组织病理无明显变化.SD-Wistar组手术成功57只,受体存活时间8～20 d,平均10.5 d;大鼠于术后3～5 d出现急性排斥反应,未经处理后均死亡. 结论 改良式肝移植操作简便,成功率高,可为大鼠原位肝移植实验提供稳定可靠的动物模型.     

豚鼠至大鼠肝移植肝下下腔静脉套管方法的改进

目的探讨豚鼠至大鼠异种原位肝移植中应用改进的肝下下腔静脉套管方法的手术效果。方法豚鼠和SD大鼠各30只分别作为供、受体,并随机配对分为实验组和对照组。实验组采用改进的肝下下腔静脉套管进行肝下下腔静脉袖套管法吻合。对照组采用常规方法进行肝下下腔静脉袖套法吻合。比较两组的供肝切取时间(切、修肝时间)、受体手术时间、无肝期时间、手术成功率、肝下下腔静脉袖套吻合口附近出血发生率以及术后生存时间。结果实验组和对照组的供肝切取时间分别为(32.2±3.5)min、(45.4±5.7)min,修肝时间分别为(14.5±2.1)min、(9.2±1.8)min,切、修肝总时间分别为(46.7±4.8)min、(54.6±6.9)min,比较差异均有统计学意义(均为P0.05)。两组受体的手术时间分别为(52.7±6.1)min、(53.2±6.5)min,无肝期分别为(16.8±2.1)min、(17.2±2.5)min,比较差异没有统计学意义(均为P0.05)。实验组与对照组的手术成功率分别为93%、53%,比较差异有统计学意义(P0.05)。两组的肝下下腔静脉袖套吻合口附近出血发生率分别为0、38%,比较差异有统计学意义(P0.05)。两组手术成功的动物的术后生存时间分别为(115±24)min、(95±29)min,比较差异无统计学意义(P0.05)。结论应用改进的肝下下腔静脉套管方法进行豚鼠至大鼠原位肝移植,手术成功率高,可用于豚鼠至大鼠原位肝移植实验研究模型的制作。     

大鼠原位肝移植模型不同灌注方式和吻合方法的效果比较

目的 分析不同灌注方式和不同肝上下腔静脉吻合方法对建立大鼠原位肝移植模型的影响。方法 将80只SD大鼠随机分为供肝获取组和受体肝移植组，每组40只。供肝获取组10只大鼠采用腹主动脉输液器滴灌法（1滴/s）,10只大鼠采用腹主动脉微量泵灌注法（6 mL/min），之后分别进行受体肝移植（每组对应10只大鼠肝移植），收集2组大鼠的供肝灌注时间和供肝获取时间，并于灌注后、肝移植24 h取肝脏组织行HE染色。受体肝移植组10只大鼠肝移植时采用连续吻合法，10只采用减张力半针吻合法（2组均在成功完成10只肝移植后截止），收集2组大鼠的肝上下腔静脉吻合时间、无肝期时间和术后发生并发症发生情况。结果 与腹主动脉输液器滴灌组比较，腹主动脉微量泵灌注组的供肝灌注时间和供肝获取时间较短（P<0.05）;HE染色结果显示，腹主动脉微量泵灌注组的肝细胞、门静脉及胆管形态学无明显改变，仅个别淋巴细胞浸润。与连续吻合组比较，减张力半针吻合组大鼠的肝上下腔静脉吻合时间和无肝期时间较短（P<0.05），术后吻合口出血和供肝灌注不全发生率较低（P<0.05）。结论 相对于腹主动脉输液器滴灌法，腹主动脉...     

受体肝分步切除三袖套法大鼠原位肝移植术的改进

目的探讨改进的三袖套法大鼠原位肝移植术的效果。方法56只成年健康Wistar封闭群大鼠,供受体各28只,采用受体肝分步切除的三袖套法行大鼠原位肝移植术。供肝置入时先不切除受体肝,而是在吻合血管的过程中分步切除受体肝。结果本组行大鼠原位肝移植28例,无肝期平均11.2(7~13)min,手术成功率92.9%(26/28)。死亡2例,1例死于出血,1例因肝上下腔静脉套管扭曲、脱落死亡。1周存活率82.1%(23/28)。结论采用受体肝分步切除的三袖套法行大鼠原位肝移植术,可缩短无肝期,提高术后存活率。     

大鼠肾移植模型手术改良技巧探讨

目的探讨大鼠肾移植模型手术的改良方法。方法供体Sprague-Dawley(SD)大鼠21只,受体Wistar大鼠42只。采用双侧供肾。受体左肾切除后借助自制导管,行受体肾动脉与供体肾动脉、受体肾静脉与供体下腔静脉端端吻合,供体输尿管带膀胱瓣与受体膀胱吻合,最后切除右肾,腹腔内注入头孢米诺10 mg,关腹。记录手术时间,动、静脉吻合时间,冷、热缺血时间等手术数据;术后大鼠存活3 d认为模型建立成功,计算建模成功率,分析死亡原因。结果供体手术时间为(32.7±5.6)min,供肾修整时间为(4.2±1.1)min。受体手术时间为(42.3±4.9)min,其中动脉吻合时间为(10.1±3.2)min,静脉吻合时间为(13.9±2.5)min,尿路重建时间为(6.3±1.4)min。热缺血时间为(5.4±1.8)s,冷缺血时间为(56.2±7.3)min。42只受体大鼠中,建模成功40只,成功率为95%。另2只受体大鼠死亡,其中1只死于血管吻合口出血,1只死于尿瘘引致的腹膜炎。结论采用改良的血管端端吻合法建立大鼠肾移植模型具有操作简单、手术时间短、成功率高的特点。     

大鼠原位节段小肠移植模型的改进

目的 建立一种简便、稳定的大鼠原位节段小肠移植模型.方法 供、受体均为雄性SD大鼠,各40只,采用供体肠系膜上动脉-腹主动脉漏斗状袖片与受体肾下腹主动脉端侧吻合,供体门静脉与受体左肾静脉Cuff套管袖套吻合.切除受体大部分小肠,供体小肠近、远端分别与受体残留小肠近、远端行端端吻合.结果 供体手术时间(40 ±5)min,受体手术时间(50 ±8) min.热缺血时间(5±2)min,冷缺血时间(15±5) min.动脉吻合时间(5±2)min,静脉吻合时间(4 ±2)min.90.0％ (36/40)的大鼠术后存活时间＞10d.结论 该模型操作简便,手术时间短,模型成活率高,稳定性好.     

大鼠心跳停搏供肝在原位肝移植术中损伤的预防

目的：探讨预防和减轻大鼠心跳停搏供肝在原位肝移植术中的损伤，以提高手术成功率。方法：雄性SD大鼠随机分为心跳停搏热缺血30min(N-30)和45min(N-45)两组;，每组分别行原位肝移植术30只次。同时，根据是否对供体手术方法进行改进又分为常规组和改良组。结果：(1)常规组和改良组的冷缺血时间分别为(70.04±1.48)和(70.36±1.42)min(P>0.05)，无肝期均为(16.40±0.73)min，肝下下腔静脉阻断时间均为(22.75±1.16)min，受体手术时间均为(90.58±3.76)min。(2)N-30和N-45常规组分别有5和9只受体术后死于原发性移植肝无功能，而改良组仅为1和2只(40%∶12%,P<0.05);(3)N-30和N-45组因术中分别出现供肝损伤致再灌注后供肝大量渗血、无肝期过长、切除受体肝脏时麻醉过深，而各有5和7，2和1，2和2只受体术后死亡。(4)N-30和N-45组术后1周存活率分别为50%和30%(P<0.05)。结论：预防心跳停搏供肝游离时损伤、供肝再灌注后渗血、无肝期过长和切除受体肝脏时麻醉过深是大鼠心跳停搏供肝原位肝移植手术成功的关键。     

猪到猴异位辅助性肝移植模型的建立

目的建立长白小家猪到恒河猴异位辅助性肝移植模型,总结手术操作要点。方法以健康雄性长白小家猪和健康恒河猴各5只建立猪到猴异位辅助性肝移植模型。以长白小家猪作为肝移植供体,以恒河猴作为受体。保留长白小家猪的右后叶和部分右前叶作为供肝,移植到受体的左肾窝和左结肠旁沟处。短暂阻断受体的腹主动脉和下腔静脉血流后,将移植肝的门静脉和肝下下腔静脉分别与受体的腹主动脉和下腔静脉行端侧吻合。结扎移植肝的肝动脉,不予重建。术后观察受体的一般情况和生存时间。结果成功建立4对肝移植模型,供肝切取时间24～35min、（30±5）min,供肝修整时间31～51min、（40±10）min,受体下腔静脉阻断时间23～36min、（30±6）min,受体腹主动脉阻断时间22～38min、（30±8）min,肝移植手术时间130～310min、（220±80）min,术中失血35～48mL、（42±6）mL。术后均无吻合口血栓形成及胆漏发生。4只受体分别于术后48、54、88及96h死亡,死亡原因均为排斥反应及术中失血过多。结论猪到猴异位辅助性肝移植模型的可重复性强、手术易操作、移植器官灌注良好,可用于猪到非人类灵长类动物肝移植的进一步研究。     

改进的门静脉套管置人技术在大鼠肝移植中的应用

目的 探讨改进的门静脉套管置入技术“门脉分支悬吊法”在大鼠肝移植门静脉重建中的应用效果.方法 “双袖套法”建立大鼠肝移植模型,分别应用改进的“门脉分支悬吊法”和传统方法行受体门静脉重建,比较门静脉重建所需时间和成功率、无肝期时间以及移植手术成功率.结果 应用改进的“门脉分支悬吊法”行大鼠肝移植(n=35),受体门静脉重建成功率94.3％,门静脉吻合所需时间仅为(1.9±0.7)min,无肝期为(21.8±2.2)min,移植手术成功率为80％,优于传统方法(P＜0.05).术后l周存活率为85.8％,与传统方法比较无统计学差异(P＞0.05).结论 改进的“门脉分支悬吊法”缩短了受体门静脉吻合时间和无肝期,提高了门静脉重建成功率和移植手术成功率,可以快速、安全地实现大鼠肝移植受体手术中的门脉套管置入和门静脉重建.     

不同术式大鼠原位肝移植模型的比较

目的比较二种不同术式大鼠原位肝移植模型的优劣,建立一种稳定的大鼠肝移植模型。方法采用二袖套法完成大鼠原位肝移植40例,采用三袖套法施行大鼠原位肝移植20例。结果二袖套法手术成功率为92．5％（37／40）,1周存活率为89．2％（33／37）。三袖套法手术成功率为85％（17／20）,1周存活率为11．8％（2／17）。结论大鼠原位肝移植二袖套法较三袖套法远期存活率高,并发症少,是一种稳定可靠的大鼠肝移植模型。     

肝硬化大鼠原位肝移植模型的制作及术后排斥反应的观察

目的 制备肝硬化大鼠的原位肝移植模型,观察术后排斥反应发生情况,为进行其它研究创建一个平台.方法 以皮下注射CCl4联合饮用苯巴比妥钠和乙醇溶液的方法制备大鼠肝硬化模型,应用改良的"二袖套"法建立大鼠原位肝移植模型,同系移植者的供、受者均为SD大鼠(SD实验组),以接受肝移植的正常SD大鼠为对照(SD对照组);同种移植者的供者为Lewis大鼠,受者为BN大鼠(BN实验组),以接受肝移植的正常BN大鼠为对照(BN对照组).术后观察受者的存活情况以及移植肝的组织学变化.结果 肝硬化大鼠门静脉压力为(182.0±10.7)mm H2O,显著高于正常大鼠的(70.8±5.5)mm H2O(P＜0.01),移植后7 d降至(82.7±10.7)mm H2O.同种移植组术后5～12 d,移植肝组织中均可见中、重度急性排斥反应病理改变.同系移植者存活时间中位数均＞100 d;同种移植者中,BN对照组和BN实验组受者肝移植后存活时间中位数均为10 d.结论 以Lewis大鼠为供者、肝硬化BN大鼠为受者制备的肝移植模型术后排斥反应的发生率较高,可作为肝移植后排斥反应相关研究的平台.     

大鼠原位肝移植模型制作过程中麻醉方法的选择

目的 研究在大鼠原位肝移植模型制作过程中麻醉对大鼠的影响，以便合理选择正确的麻醉方法。方法 用SD大鼠以改进的二袖套法制作大鼠原位肝移植模型。将100只大鼠随机分为5组：对照组，乙醚组，氯胺酮组，水合氯醛组，戊巴比妥组。观察每种麻醉剂的麻醉过程、对大鼠生理和肝功能的影响以及无肝期开始后的死亡率。结果 每种麻醉剂的麻醉过程不尽相同。麻醉对大鼠生理均有影响，其中以乙醚的影响较小，戊巴比妥有较大的肝毒性。无肝期开始后氯胺酮组、水合氯醛组、戊巴比妥组死亡率较高，而乙醚组无死亡发生。结论 各种麻醉对肝移植大鼠均有明显影响。乙醚具有对生理、肝功影响小，无肝期开始后死亡率低的优点，可作为制作大鼠原位肝移植模型的首选麻醉方法。     

血管活性物质在肝硬化肝移植后高血流动力学中的作用

目的：探讨内源性血管活性物质在肝硬化鼠肝移植后设备在流动力学中的作用,方法：雄性SD大鼠随机分为四组,正常对照组（NL）,肝硬化组（IHPH）,正常鼠肝移植组（NL－OLT）和肝硬化地移植组（IHPH－OLT）,IHPH－OLT鼠又分为术后3天（A组）和7天（B组）两个亚组,IHPH模型肌注CCI4制备,大鼠OLT模型采用三袖套法,血流动力学研究采用放射性微球注射技术,血浆胰高糖素（Glu),一氧化氮（NO）,前列环环素（PGI2）,血栓素（TXA2）以及内皮素（ET）浓度用放射免疫法测定,结果：NL－OLT鼠绝大多数血流动力学参数与NL鼠比较差异无显著意义（P＞0．05）,与NL鼠比较,NL－OLT鼠的NO和PGI2均无明显变化,而Glu,ET 和TXA2水平显著地升高（P＜0．05）,IHPH,IHPH－OLT A,B鼠均具有全身和内脏高血流动力学特征,其高动力循环的程度和血管扩张物质NO和Glu增加的程度均是IHPH＞IHPH－OLTA＞IHPH－OLTB鼠的PGI2显著地高于NL鼠,IHPH－OLTA、B鼠的PGI2显著地低于IHPH鼠（P＜0．05）,而HPH－OLTB鼠与NL鼠比较已无显著差异,血管收缩物质ET和TXA2在肝硬化肝移植术后均有不同程度升高,结论：肝移植术本身并不导致术后高血流动力学的发生,血管扩张物质NO和Glu,尤其是NO在IHPH以及IHPH－OLT鼠的高动力循环中起重要作用,肝硬化肝移植后早期依然存在的高动力循环是术前引起高血流动力学发病因子升高的原因未消除的结果。     

Comparison of two techniques of arterial anastomosis during adult cadaveric liver transplantation

Arterial complications are a major source of morbidity and mortality after orthotopic liver transplantation (OLT). The incidence of hepatic artery thrombosis (HAT) ranges from 1.6% to 8%, with a mortality rate that ranges from 11% to 35%. We have described herein a technique of arterial anastomosis aiming to perform the anastomosis as straight as possible to avoid any kinking, redundancy, or malposition of the artery when the liver is released in its final position. We compared this technique with the traditional technique of arterial anastomosis using an aortic Carrel patch, namely, 198 OLT (group A) with the traditional technique and 117 OLT (group B) with the modified technique. An aorto-hepatic bypass was necessary in 25% of the cases in group A and in 21% of the cases in group B (P = .33). Vascular anomalies were present in 20% of cases in group A and in 27.5% in group B (P = .14). Fourteen cases (7%) of HAT developed in group A versus 0 cases in group B (P = .003). In group B, we experienced 2 (1.7%) late arterial stenoses that were successfully treated using percutaneous transluminal angioplasty. The 14 cases of HAT occurring in group A were successfully managed using immediate surgical revascularization with graft salvage in 6 cases (43%), whereas the remaining 8 cases needed urgent retransplantation. We suggest that a technique of arterial anastomosis aimed at avoiding kinking, redundancy, or malposition of the artery may be a viable option to reduce the risk of HAT after OLT.     

Protocol for the technique of orthotopic liver transplantation in the rat

Orthotopic rat liver transplantation (OLT) has been generally accepted as an excellent model for the analysis of pathological, physiological, and immunological aspects related to organ transplantation. However, many researchers require a long training period to achieve a high success rate using this major surgical procedure on small animals. We therefore developed a protocol for learning rat OLT. It is recommended, initially, that the heterotopic heart transplantation (HHT) model be used to master the cuff technique, since this technique is similar to that in OLT and leads to an understanding of another organ transplantation technique. For beginners who advance beyond the HHT step to OLT but cannot finish the anhepatic phase within 30 min, we recommend the use of portosystemic-shunted rats as recipients. These animals have had their spleens transposed subcutaneously more than 3 weeks before use. The use of these modifications and this training program makes it possible to master the techniques and to achieve a high success rate with a short training period. © 1993 Wiley-Liss Inc.     

Experiences in liver transplantation for hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. Beside surgical resection, orthotopic liver transplantation (OLT) is not only effective but also the only potentially curable treatment in selected cases of small tumors. We report our experience in 11 male patients transplanted for HCC from August 1998 to July 2002. Selection criteria for OLT were unresectability of the hepatic tumor and severity of the underlying liver disease. The tumor diagnosis was confirmed by histology, imaging techniques, and tumor markers. All patients received an orthotopic liver allograft using a modified piggyback technique. Six of the 11 patients are alive; one died due to acute rejection and four died from recurrent disease. In all four patients with recurrent disease, vascular invasion was shown histologically, whereas only one patient without evidence of recurrence showed vascular invasion. To prevent recurrence after OLT the immunosuppressive regime was adjusted to the underlying disease by early cessation of prednisolone and reduction in the long-term exposure to immunosuppressive drugs. Patients were screened for recurrence by ultrasound and computed tomography. Recurrent HCC were treated symptomatically. OLT is an effective treatment for subgroups of patients with HCC. It might be possible to downstage the liver tumor by chemoembolization and/or radiofrequency ablation and allow the patients to wait for a suitable donor. After OLT the early withdrawal of prednisolone and the reduction of other immunosuppression is feasible. In conclusion, OLT can be a potentially curative therapy for HCC.     

Choice of transplantation techniques and indications for liver transplantation in polycystic liver disease in patients with no signs of end-stage liver disease

OBJECTIVE: Since the initiation of the Liver Transplant Program, 500 liver procedures have been performed. Polycystic liver disease (PLD) and polycystic kidney-liver disease (PKLD) have been rare indications for orthotopic liver transplantation (OLT). Only 7 patients (1.4%) underwent transplantation due to PLD and PKLD. MATERIALS AND METHODS: The group consisted of 4 patients who underwent OLT (0.8%) and 3 patients who received simultaneous liver kidney transplantation (LKT; 0.6%). Our objective was to analyze the indications for either OLT or combined LKT as well as indications for surgical techniques during OLT among patients with PLD or PKLD. RESULTS: The main indication for OLT was massive hepatomegaly causing severe physical handicaps, fatigue, and clinically advanced malnutrition. All 3 patients with indications for combined LKT were dialysis-dependent. None of the patients had symptoms of end-stage liver disease and/or hepatic failure. In 4 cases, a portal bypass was applied, and the piggy-back method used in the other 3 cases. The hepatectomy caused no uncommon difficulty. In cases of simultaneous transplantations, the kidney was implanted separately after OLT. All patients are alive following the transplantation; major surgical complications have occurred. CONCLUSIONS: Patients with PLD can undergo OLT safely with good results. They benefit from the relief of abdominal distension and anorexia. Patients with PKLD who are dialysis-dependent should undergo simultaneous LKT. The surgical technique was solely dependent on the intraoperative conditions determined during the dissection phase.     

Rat liver transplantation for total vascular reconstruction, using a suture method

We developed a novel protocol for rat orthotopic liver transplantation (OLT), using a suture method to establish hepatic artery flow. After determining that early inferior vena cava (IVC) unclamping maintained better circulation compared with the portal vein (PV) using porto-systemic shunted recipients, we developed a rat OLT model with total vascular reconstruction using a suture method. After connecting the suprahepatic IVC, the infrahepatic IVC was anastomosed, using a running suture method. IVC circulation was established immediately. The PV was anastomosed without intestinal congestion, using porto-systemic shunted recipients. The aortic conduit, including the donor celiac and hepatic artery, was anastomosed to the recipient abdominal aorta end-to-side. Eight of 11 OLT cases (72.7%) survived indefinitely. Biliary connection was achieved using a one-stent method. Three cases died 3-5 days postoperatively. Hepatic angiography showed good patency. The graft liver was histologically normal in long-surviving rats.