小鼠异位小肠移植模型的建立

目的 探讨稳定的小鼠异位小肠移植模型制作方法,为小肠移植排斥反应的研究提供良好的实验工具.方法 选用C57BL/6小鼠作供体和BALB/c小鼠作受体进行同种异基因型异位节段性小肠移植.采用小肠供体的门静脉与受体下腔静脉端侧吻合,供体带主动脉片的肠系膜上动脉与受体腹主动脉端侧吻合,供体近端肠管结扎,远端与受体空肠端侧吻合的方式建立异位小肠移植.术后禁食3天,不禁饮,每天分两次经皮下分别给予5%葡萄糖生理盐水2 mL,术后不使用抗生素和免疫抑制剂.小鼠存活超过5 d视为手术成功.结果 共行小肠节段性移植30例,术后5 d存活率达70%(21/30).供体手术时间(41±5.5)min,热缺血时间约0.5 min,供体肠段肠系膜上动脉组织片修整时间约为3 min,供体冷保存时间为(30±7.5)min,受体手术时间(90±7.5)min,其中腹主动脉及下腔静脉阻断时间为(40±3.0)min,静脉吻合时间(10±2.0)min,动脉吻合时间(15±2.5)min,成活小鼠受体手术平均出血量约0.2 mL.手术失败的9例小鼠的死亡原因为动脉吻合口部位狭窄及吻合口处血栓形成(6例),吻合口出血导致出血性休克(2例)和术后腹腔内感染(1例).结论 良好的供体肠段的获取、高质量的血管吻合和肠道吻合及供、受体补液是提高小鼠小肠移植手术成功率的关键.     

大鼠原位小肠移植模型的建立与改进

目的探讨建立一种简便稳定存活率高的大鼠原位小肠移植模型。方法整块切取带有腹主动脉和肠系膜上静脉并门静脉的节段小肠,术中原位冷灌注,4℃乳酸林格液保存。动脉吻合采用显微外科技术行供体腹主动脉对受体腹主动脉的端侧吻合,利用Cuff套管技术将供体的门静脉与受体的左肾静脉端端吻合。移植肠远、近端分别与受体肠行端端吻合。结果建立小肠移植模型1 6次,动脉、静脉吻合时间分别为(2 5±5)m in和(4±1)m in。1 6只受体鼠中1 3只存活超过5 d,平均存活(1 0.3 5±2.8 4)d,最长存活时间为2 1 d。结论移植肠的获取、血管吻合技术、肠吻合技术和维持良好的血容量是手术成功的关键。该模型的成功建立,为小肠移植的基础研究提供了良好的动物模型。     

大鼠原位肾脏移植模型的显微外科技巧

目的 探讨建立稳定、可靠的大鼠原位肾脏移植模型的显微外科手术技巧。方法供体手术经腹主动脉以4℃乳酸钠林格注射液对供肾进行原位灌洗。受体手术采用将供肾的血管与受体的同名的血管作端端吻合的大鼠原位左肾移植。结果 共施行大鼠原位肾脏移植手术120例，手术成功率为91．5％；非干预组中，同系移植大鼠1个月存活率为82．9％。结论 肾动、静脉和输尿管的重建均采用端端吻合法而无须阻断腹主动脉和下腔静脉，大大减少了对受体循环系统的影响。该模型稳定性强、重复性好，适合于移植免疫的基础研究。     

大鼠肾脏移植输尿管吻合方法的改进

目的 报道一种改进的大鼠上段输尿管端端吻合技术应用于大鼠的肾脏移植模型.方法 以封闭群SD大鼠作为供受体.按常规行肾动脉、肾静脉的端端吻合,开放血流后进行尿路重建.改进的大鼠上段输尿管显微吻合方法:吻合口靠近受体输尿管上端肾盂处;根据受体输尿管口内径剖开供体输尿管的远肾下极缘;采用暂时性内支架辅助的间断缝合确保缝合质量.结果 使用改进的显微输尿管吻合方法进行原位大鼠肾脏移植30例,近期发生动脉血栓、尿漏、动脉吻合口出血各1例.术后7 d大鼠存活90%;术后21 d,显微镜下解剖输尿管吻合口,发现部分梗阻移植肾积水1例;尿路并发症总的发生率6.67%.结论 改进的输尿管吻合方法简便、可靠,降低了技术难度.易操作,相关并发症的发生率较低,能够在10倍的放大视野下完成.     

大鼠小肠移植模型的改进

目的　通过改进技术 ,建立一种简便稳定存活率高的大鼠异位节段小肠移植模型。方法　“无损伤”游离 ,原位冷灌注 ,切取带有腹主动脉和肠系膜上静脉并门静脉的节段小肠 ,4℃乳酸林格氏液保存 1h。游离受体左肾静脉 ,切除左肾。采用显微外科技术行供体腹主动脉对受体腹主动脉的端侧吻合 ,门静脉与受体左肾静脉行袖式吻合。移植肠近端关闭 ,远端外置。结果　共进行 87次移植实验 ,其中 2 6次为正式实验。动脉、静脉吻合时间分别为 2 5～ 30min和 5min。 2 6只受体鼠中2 1只存活超过 3d ,平均存活 (8.93± 2 .5 9)d ,最长存活时间为 14d。结论　良好的血管吻合和充分补充液体是手术成功、移植肠具有良好活力的关键因素     

异基因大鼠胰十二指肠移植模型建立的手术技巧

目的 探讨异基因大鼠全胰十二指肠移植(WPDT)模型手术过程的操作要点.方法 将供、受体腹主动脉端侧吻合,供体门静脉与受体左肾静脉袖套吻合,供体十二指肠与受体小肠端侧吻合,成功建立Lewis→Wistar糖尿病大鼠异基因WPDT模型.结果 50只糖尿病大鼠行WPDT术式,44只移植成功.其中8只于术后3 d内死亡,其余36只受体大鼠存活时间为6～16 d,平均为(10.45±3.30) d,术后7～10 d为死亡高峰,移植物病理改变呈典型的急性排斥反应. 结论 熟练的显微技术和重视操作细节是成功建立WPDT模型的关键因素.     

肠道-腔静脉引流的胰肾联合移植术

目的 探讨肠道-下腔静脉引流的胰肾联合移植术的手术操作及临床效果.方法 对3例慢性肾衰竭合并2型糖尿病患者施行肠道-腔静脉引流的胰肾联合移植术,3例均为首次移植,年龄52、58、58岁.每日胰岛素用量20～55 U.供体切取均采用多器官联合切取.热缺血时间8～12 min,供体修整均采用肝总动脉与胃十二指肠动脉端-端吻合以重建十二指肠动脉弓,利用供体髂总静脉延长供体门静脉,切除供体脾脏.以供体髂内动脉与供肾动脉端-端吻合备用.受者手术采用右下腹经腹直肌切口,游离腔静脉下段及右侧髂外动静脉,取动脉延长之供肾,将供肾静脉与受者髂外静脉行端-侧吻合,将供体髂总动脉与受者髂外动脉行端-侧吻合,供体髂外动脉(残端修整成斜面)以动脉夹暂时夹闭备用,十字切开侧腹膜,将供肾埋入,输尿管经腹膜外隧道牵至膀胱底行膀胱输尿管吻合术,胰腺移植采用供体门静脉与受者下腔静脉行端-侧吻合,腹腔干-肠系膜上动脉之腹主动脉袖片与供体髂外动脉残端吻合,开放血流后行供体十二指肠与受体小肠侧侧吻合并关闭十二指肠残端.术后保留胃肠减压,待患者胃肠道功能恢复后拔除.每4 h测血糖、每6 h测血清及胰周引流液淀粉酶1次,每日超声监测胰腺及肾脏血流,生长抑素0.1 mg皮下注射8 h 1次,2周后停用.免疫诱导采用抗胸腺细胞免疫球蛋白减激素方案.结果 3例患者手术过程顺利,手术时间分别为7.5、8.0及10.0 h,术中失血量300～500 ml,仅1例术中输注浓缩红细胞2 U.术后1～3 d内完全停用胰岛素.术后3～7 d内移植肾功能恢复正常.实验室检查SCr分别为86、98及112μmol/L.1例术后10 d出现消化道出血,考虑为肠道吻合口出血;停用抗凝药.给予止血药及输血6 U治疗后1 d出血停止.3例随访2～6个月,无排斥反应发生,空腹及餐后血糖正常.结论 利用供体髂动脉搭桥的方法进行的胰肾同侧联合移植术手术操作简单,创伤较小而且仪使用一侧髂血管,对于左侧髂动脉硬化严重的患者仍可施行该术式.因而扩大了受者的范围;为患者保留一侧髂血管,为今后再次肾移植创造了条件.同时腔静脉引流的胰肾联合移植术使供体门静脉与脾静脉的夹角更符合生理角度,可能减少脾静脉血栓形成的发生率.     

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.     

猪胰肾联合移植模型的建立

目的 建立一种猪胰肾联合移植模型。方法 血管吻合采用供体门静脉远端与左肾静脉端端吻合，近端再与受体下腔静脉端侧吻合；带腹腔动脉、肠系膜上动脉和左肾动脉的腹主动脉段与受体腹主动脉端侧吻合。胰腺外分泌自膀胱引流，供肾输尿管置管引外流。结果 １１例动物移植成功，早期移植功能恢复良好。结论 这种建立猪胰肾联合移植的方法是可行的。     

经门静脉及肠道引流式一期胰肾联合移植六例报告

目的介绍6例门静脉肠道引流胰肾联合移植术及其疗效。方法对6例胰岛素依赖性型糖尿病合并尿毒症患者施行了同期门静脉肠道引流胰肾联合移植术,即带腹腔动脉和肠系膜上动脉开口的Carrel袖片与髂外动脉行端侧吻合,门静脉与受体肠系膜上静脉行端侧吻合,移植物十二指肠与受体空肠行侧侧双层吻合。术后早期采用肾上腺皮质激素 霉酚酸酯 他克莫司 巴利昔单抗四联诱导治疗。结果6例胰肾联合移植手术均获成功,4例存活至今,2例在围手术期死亡。4例存活者术后3d空腹血糖恢复正常;停用胰岛素后,肾功能3~5d恢复正常;血淀粉酶和血清C肽水平2周后均在正常范围。随访4~34个月,移植胰和移植肾功能正常。2例死亡的原因是胰漏感染和FK506药物中毒。结论门静脉肠道引流方法更符合生理,无远期并发症,是很有前途的胰腺移植手术方式。     

大鼠胰十二指肠肾脏联合移植模型的实验研究

目的：为了进行胰肾联合移植基础研究，建立一种简易可靠的大鼠胰十二指肠肾脏联合移植（SPKT）模型，方法：雄性SD大鼠作同品系异体移植的供受体，受体尾静脉注射链脲霉素（STZ）50mg/kg，建立大鼠糖尿病模型，在保存液中，移植物门静脉与肾下下腔静脉作袖套式吻合；移植物肾上腹主动脉，肾上下腔静脉分别与受体肾下腹主动脉，肾下下腔静脉显微缝合吻合，移植物十二指肠与受体十二指肠端侧吻合，移植物带输尿管的膀胱片与受体膀胱吻合。结果：血管吻合时间及受体手术时间短，手术成功率为88％，移植胰腺及肾脏功能良好。结论：此模型是一种简易可靠的大鼠胰十二指肠肾脏联合移植模型。     

Technique for retransplanting heterotopic heart grafts in mice

Removal of a transplanted organ from its original recipient and retransplanting it into a new host is an important method to study the role of the graft in the rejection process. Here we describe a novel technique of heart retransplantation in the mouse. In this technique, a primarily vascularized heart graft is anastomosed to the abdominal aorta and inferior vena cava of a syngeneic or immunodeficient allogeneic mouse, using standard techniques. Either 10 or 70 days later, the same graft is retransplanted into the abdomen of a second mouse by end-to-side anastomosis of the donor (first recipient) aortic and inferior vena cava's cuffs to the second recipient's abdominal aorta and inferior vena cava, respectively. A greater than 90% success rate was achieved by using this microsurgical technique. This method should be useful for studying intragraft factors, such as ischemia-reperfusion injury and donor antigen-presenting cells, on the outcomes of transplantations.     

Surgical Techniques of Allogeneic Liver Transplantation in a Nonhuman Primate Model

Herein, we report our experience of performing allogeneic orthotopic liver transplantation (LT) in nonhuman primates. We designed an allogeneic ABO-compatible orthotopic LT model in monkeys in a manner similar to that used in humans. We applied almost the same surgical procedures used for human conventional deceased donor LT. A total of 6 monkeys underwent allogeneic LT. One cynomolgus monkey aged 45 months (3.4 kg) and 5 rhesus macaque monkeys aged 50.2 ± 14.8 months (5.40 ± 0.33 kg) were used as recipients. In the donor surgery, the liver was perfused in situ through the aorta using cold histidine-tryptophan-ketoglutarate solution. The portal vein (diameter, 5–10 mm), supra- and infra-hepatic inferior vena cava (IVC) (diameter, 12–15 mm), and common bile duct (diameter, 1.5–3.0 mm) were dissected out. The hepatic artery was kept in continuity with the celiac trunk and abdominal aorta up to the iliac bifurcation (diameter, 5–6 mm). The mean graft weight was 102.0 g (94.8–111.0 g). Recipient surgery was conducted in parallel. After recipient hepatectomy, the graft was implanted. The suprahepatic IVC and portal vein were anastomosed to those of the graft. After reperfusion, the infrahepatic IVC was anastomosed. The aorta conduit of the graft was anastomosed to the infrarenal aorta of the recipient in a retrocolic end-to-side manner. Biliary reconstruction was performed in a duct-to-duct anastomosis with cholecystectomy. Mean operative time was 107.0 minutes for donor and 198.2 minutes for recipient. There was one operative death due to unknown cause. In conclusion, for allogeneic orthotopic LT in nonhuman primate model, we can apply almost the same procedure used for human conventional deceased donor LT in a similar manner.     

简单条件下大鼠肾移植模型的建立

目的探讨在简单条件下,稳定的、用于移植后早期免疫功能等实验研究的大鼠肾移植模型的建立方法。方法以Wistar大鼠为供体,Sprague Dawley大鼠为受体。取供体大鼠左肾,移植物包括与肾静脉相连的下腔静脉段,与肾动脉相连的腹主动脉段,以及与输尿管相连的供体膀胱瓣,经腹主动脉原位低温灌注6～8mL4℃肝素生理盐水。受体手术于裸眼下完成,供体下腔静脉与受体下腔静脉、供体腹主动脉与受体腹主动脉行端侧吻合,供体输尿管带膀胱瓣与受体膀胱两定点连续缝合。结果共完成50例异体肾移植大鼠模型,存活41例,手术成功率82%,存活时间（6.3±1.6）d。供体手术时间（44.8±7.4）min、受体手术时间（59.0±6.6）min、动脉吻合时间（15.9±2.3）min、静脉吻合时间（14.2±2.7）min、尿路重建时间（5.3±0.8）min、热缺血时间（55.7±4.5）s和冷缺血时间（55.1±5.9）min。结论建立此模型所需要的实验条件简单,术者容易掌握,移植成功率高。     

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.     

Orthotopic knee grafts in rats: a model for growth plate transplantation

A model for orthotopic whole knee transplantation in syngeneic Lewis rats is presented. The donor knee was isolated on the femoral vessels 1 cm superior and inferior to the knee joint. A minimal muscle cuff was left attached to ensure adequate circulation to the graft via the popliteal vessels. The graft was transplanted to the recipient animal and secured in an orthotopic position by two intramedullary Kirschner wires. The graft pedicle was anastomosed to the recipient's femoral vessels and the muscular elements sutured. Preliminary results are reported, and future implications on growth plate autograft transplantation are discussed.     

Modified technique for kidney transplantation in mice.

This study describes a new method for joining the donor ureter to the recipient bladder during mouse kidney transplantation. The donor left kidney was harvested using methods previously published, except that bladder tissue was not harvested with the end of the ureter. The recipient left kidney was removed and the donor kidney was attached using end-to-side anastomosis. The recipient bladder was pierced with a 21-gauge needle allowing curved forceps to be inserted through the bladder, to pull through the ureter, and the periuretal tissue was stitched to the exterior wall of the bladder. The donor ureter was allowed to retract inside the bladder. Following a right nephrectomy, grafts were monitored by blood serum creatinine and urea. With a technical success rate of 83%, this technique reduced donor harvest time by 20 minutes and ureter attachment time by 15 minutes making it the best method available for mouse kidney transplantation.     

14例亲属活体供肾动脉变异的血管重建

目的 探讨亲属活体供肾动脉变异的血管重建方法.方法 在104例亲属活体供肾移植中,有14例供肾动脉变异.供肾动脉变异的分类和血管重建方法分别为:(1)单支动脉较早分支型2例,取肾时分支受损,分别用受者髂内动脉及其分支、腹壁下动脉离体重建受损动脉.(2)双支动脉型10例,4例用受者髂内动脉及其分支离体重建血管,3例用受者腹壁下动脉与较细分支于体内吻合,1例较短肾动脉与较长肾动脉端侧吻合,1例较细副.肾动脉与主肾动脉端侧吻合,1例双支分别与髂外动脉端侧吻合.(3)3支动脉型2例,1例用受者髂内动脉及分支离体重建血管,1例结扎细小分支后,将较细的副肾动脉与主肾动脉端侧吻合.14例血管重建后,分别将供肾动脉较粗支和/或髂内动脉主干端与受者髂外动脉端侧吻合.结果 术后各支动脉血流通畅,移植.肾血液供应丰富、均匀.12例肾功能早期恢复正常,其中1例术后第14天发生急性排斥反应.1例术后即发生急性排斥反应;1例血肌酐下降缓慢.随访至2008年7月,除1例动脉粥样硬化较重的受者(三支动脉)下极动脉栓塞,血肌酐升高并稳定在170μmol/L外,其余患者动脉血流通畅,血液供应丰富、均匀.结论 供肾动脉变异时,利用所得供肾动脉的自身条件重建血管,或用受者髂内动脉及分支或腹壁下动脉重建血管,可获得较好的移植肾功能.受者动脉粥样硬化较重,同时有较细肾动脉支做重建吻合时,应注意该支动脉发生栓塞的可能.     

A rare case of complex vascular reconstruction in liver transplantation.

Abnormalities of recipient or donor vascular structures are associated with reconstructive difficulties in liver transplantation. A patient with thrombosis of the right hepatic vein and associated stricture of the inferior vena cava (IVC), portal vein thrombosis and multiple aberrant arteries underwent orthotopic liver transplantation. The donor's suprahepatic IVC was anastomosed to the recipient's intrathoracic IVC. The portal vein flow was restored by venous graft interposition, while the arterial flow was ensured by interposing an iliac arterial graft anastomosed to the infrarenal aorta. In conclusion, graft function remains excellent more than 5 years postoperatively.     

再次肾移植受者和移植肾长期预后影响因素分析

目的探究再次肾移植受者和移植肾存活情况及长期预后影响因素。 方法回顾性分析1991年1月1日至2017年12月31日于浙江大学医学院附属第一医院肾脏病中心接受肾移植受者临床资料。共纳入再次肾移植受者37例,首次肾移植受者5 374例。根据再次肾移植受者移植肾存活时间长短,将其分为长期存活组(19例,>5年)和短期存活组(18例,≤5年)。采用成组t检验比较长期和短期存活组供受者年龄、首次与再次肾移植间隔时间、HLA错配数和再次移植供肾冷/热缺血时间。采用卡方检验比较长期和短期存活组受者性别、再次移植供肾类型、再次移植前后群体反应性抗体阳性比例、首次移植失功移植肾切除比例、再次移植前免疫诱导比例及再次移植后移植肾功能延迟恢复(DGF)和急性排斥反应发生比例。采用Kaplan-Meier法分析再次和首次肾移植受者/移植肾1、5和10年存活率。采用Cox比例风险模型分析影响再次肾移植术后移植肾长期存活影响因素。P<0.05为差异有统计学意义。 结果截至2018年3月1日,37例再次肾移植受者中位随访时间为152个月(11～323个月),2例死亡,18例发生移植肾失功,17例移植肾功能稳定。5 374例首次肾移植受者中位随访时间为108.9个月(0.1～350.0个月),459例死亡,1 343例发生移植肾失功。再次移植组受者/移植肾1、5和10年存活率分别为86%/81%、86%/62%和82%/36%,首次移植组受者/移植肾1、5和10年存活率分别为99%/98%、93%/89%和88%/80%。再次移植组移植肾1、5和10年存活率均低于首次移植组(χ²＝60.816、25.110和43.900,P均<0.05);再次移植组受者1年存活率低于首次移植组,差异有统计学意义(χ²＝40.409,P<0.05)。长期和短期存活组受者再次移植后移植肾DGF和急性排斥反应发生比例差异均有统计学意义(χ²＝4.039和4.748,P均<0.05)。Cox回归分析结果示DGF和急性排斥反应是影响再次肾移植受者移植肾长期存活的独立危险因素,差异有统计学意义(RR＝4.317和4.571,P均<0.05)。 结论再次肾移植受者移植肾存活率低于首次肾移植受者,DGF和急性排斥反应是影响再次移植受者移植肾存活的独立危险因素。