大鼠肾移植慢性排斥反应模型的建立

目的建立稳定而可靠的大鼠肾移植慢性排斥反应模型。方法选用30只Wistar大鼠为供体,30只SD大鼠为受体。取供体左肾,采用HC-A离体肾保存液原位灌注,将供肾动、静脉分别与受体腹主动脉、下腔静脉行端侧吻合,以输尿管膀胱植入法行尿路重建,建立大鼠同种异体肾移植模型。分别于术后3、6、9周取移植肾观察大体和组织形态学变化,观察术后并发症及排斥反应情况。结果移植肾脏大体和组织形态学呈渐进性变化,至术后9周可出现明显的慢性排斥反应病理改变。移植肾脏可顺利存活,部分出现肾积水并发症,但不影响排斥反应病理变化。结论本方法可建立稳定、可靠的肾移植慢性排斥反应模型,是研究慢性排斥反应的理想模型。     

硬膜外导管在大鼠肾移植血管吻合中的应用

目的建立一种手术难度低,成功率高的大鼠肾移植模型。方法 Wistar大鼠作供体,SD大鼠作受体,将供体腔静脉与受体肾静脉端端吻合,供体腹主动脉与受体腹主动脉端侧吻合,供体膀胱瓣与受体膀胱吻合。根据血管吻合时应用硬膜外导管与否,将受体分为有支架组和无支架组两组。结果有支架组共进行肾移植30次,成活26只;无支架组共进行肾移植20次,成活10只。有支架组的成活率86.7%（26/30）较无支架组50.0%（10/20）明显提高（P〈0.05）,血管吻合总时间（22±2）min较无支架组（32±2）min明显缩短（P〈0.05）。结论硬膜外导管应用于大鼠肾移植血管吻合,降低了手术难度,减少了吻合口出血,提高了手术成功率。     

大鼠心脏移植急性排斥反应动物模型的建立

目的为探索器官移植后急性排斥反应的一种快速可靠的诊断方法。方法先暴露和游离受体的吻合段血管,再获取供心以缩短移植心脏冷缺血时间;保留供者的心脏及右上肺,将其胸主动脉与受者的腹主动脉间断端侧吻合;开放血流时,先开放远心端再间断开放近心端,同时按摩心脏至心脏搏动规律、有力,其色泽恢复红润,右上肺亦充盈良好。结果A组(n=20)成功16只,动脉吻合口出血2只、心脏复跳失败1只、吻合口狭窄1只;B组(n=20)成功17只,2只死于吻合口出血,1只死于吻合口狭窄。A组手术成功率为80%,B组为85%(P>0.05)。结论(1)先暴露并游离好受体血管吻合段,再获取供心,可以缩短移植心冷缺血时间。(2)边灌注边获取的方法利于快速而又完整的获取供心。(3)在切取和植入的过程中一定要注意低温保护。(4)利用腹主动脉段进行移植,使手术方便易行。左心做功的大鼠腹部心脏移植模型简单、安全、实用,因其左心参与循环而使血流动力学更接近生理状态,是进行心脏移植及血流动力学研究的较理想模型。     

大鼠异位工作型心脏移植模型的建立

目的 建立大鼠腹腔异位工作型心脏移植动物模型.方法 Wistar大鼠30只,将供心主肺动脉与左房吻合,再将供心移植到受体腹部,将升主动脉与供心腹主动脉端侧吻合,使得左心房获得前负荷,成为工作型心脏.结果 成功建立大鼠腹腔异位工作型心脏移植模型,15例成功11例,手术成功率为73%.手术平均时间为(38.8±5.7)min.移植心脏获得长期存活,在28 d后通过超声观察可见供体心脏工作正常.结论 手术的关键是提高供心的摘取技术及受体的血管吻合技术,同时术后给与供心适当的按摩辅助也非常必要.此模型可以更好的用于移植免疫学的研究.     

食蟹猴肾移植模型的建立

目的建立食蟹猴的肾移植模型。方法通过显微外科手术对食蟹猴进行同种异体移植,将供体肾动脉、肾静脉分别端侧吻合到受体腹主动脉、下腔静脉,供体输尿管端端吻合到受体输尿管。结果154例肾移植食蟹猴无一只在手术过程中因意外死亡;无一只因手术感染死亡;无一只因手术并发症死亡;肾热缺血时间（30±4.5）min。结论应用食蟹猴建立肾移植模型,技术成熟,方法可靠。应用此模型评价新型免疫抑制药物,进行免疫耐受、异种移植研究可为临床试验提供更为准确可靠的依据。     

单人直视下改良建立大鼠原位肝移植模型的体会

目的:建立稳定大鼠原位肝移植模型,缩短术中无肝期时间,提高手术成功率及受体存活率。方法:在Kamada"二袖套法"的基础上改进,单人直视下建立大鼠原位肝移植模型,行60例SD大鼠原位肝移植手术。本研究简化供受体麻醉方式,供肝采用经门静脉(必要时配合腹主动脉补救方式)进行冷灌注,缩短修肝时间,提前预置牵引线,固定进针位置,改进植入肝脏肝上下腔静脉吻合、肝下下腔静脉及门静脉套管。观察并记录各组大鼠供体手术、修肝套管、无肝期、受体手术及肝移植手术总时间。术后检测1,7,30天受体大鼠肝功能(血清丙氨酸转氨酶(ALT),天冬氨酸转氨酶(AST)及总胆红素(TB))并分析生存情况。结果:无肝期结束后,供体肝脏灌注良好,受体麻醉移除后较快苏醒。供体手术、修肝套管、无肝期、受体手术及肝移植手术总时间分别为(32.5±1.58)、(7.3±1.43)、(15.6±2.62)、(53.2±3.74)、(108.5±2.34)min。大鼠术后24 h(手术成功率)为95%,1周生存率分别为90%,1月生存率分别为86.7%。大鼠术后短时间内肝功能水平增高,24 h时ALT(228.5±54.5 IU/L),AST(439.3±86.3 IU/L),TB(6.2±0.7μM),1周后逐渐恢复正常。结论:改良后的方法可以简易麻醉流程,缩短无肝期,提高手术成功率及受体的生存率。     

小型猪胰腺移植急性排斥反应模型的实验研究

目的　建立小型猪胰腺移植动物模型 ,探索早期诊断急性排斥反应的方法。方法　 4 0只猪随机配对行胰腺移植 2 0次 ,将供胰所带的血管与受体髂血管吻合 ,所带小段十二指肠与空肠吻合 ,术中监测平均动脉压、中心静脉压及血气。术后测定受体的血淀粉酶、血糖 ,监测外周血免疫指标 ,彩色多普勒检测供胰血流及超声引导下活检、组织病理检查。结果　移植手术成功率为 90 % ,受体平均动脉压在吻合血管开放后有明显下降 ,与血流开放前差异有显著性 (P <0 0 5 ) ,输血有助于手术成功 ,受体术后平均存活 (12 6± 2 3)d。外周血免疫学监测指标早于供胰的组织病理改变 ,两者的改变均早于急性排斥反应的临床表现。结论　小型猪适用于胰腺移植模型的建立 ,加强术中循环功能的管理、及时输血有利于受体成活 ;超声引导下穿刺活检供胰的组织病理学检查与监测外周血免疫指标均适于早期诊断急性排斥反应     

大鼠肝移植急性排斥反应模型的建立与评价

目的：建立大鼠肝脏移植急性排斥反应模型,并对所建模型进行评价。方法：采用改良Kamada“二袖套”法,以近交系大鼠Dark Agouti（DA）为供体、Lewis（LEW）为受体（A组）建立大鼠原位肝移植急性排斥反应模型,通过观察手术情况、生存情况及肝功能和病理学检查对此模型进行评价,同时以LEW→LEW作为对照组（B组）。结果：手术成功率为91.3%;手术时间为（90.70±5.68）min;无肝期时间（9.96±1.19）min;平均存活时间A组为（12.44±3.43）d,B组超过100d;A组血清谷丙转氨酶、谷草转氨酶及血清总胆红素术后不断升高,在第10～14d最为明显,血清白蛋白在术后第3d开始逐渐降低,在相同时相点,与B组相比差异显著;A组移植肝脏病理检查有明显的排斥反应,而B组没有。结论：DA→LEW为稳定、强烈的大鼠肝移植急排模型,是研究肝移植排斥及免疫耐受的理想动物模型。     

大鼠腹腔内工作型心脏移植模型的改进

目的构建大鼠腹腔内工作型心脏移植模型,总结影响模型成功率的因素。方法 Brown Norway到Lewis大鼠心脏移植90例,其中预实验50例,正式实验40例。采用肺动脉和左房吻合、主动脉和受体腹主动脉吻合的方法建立工作型心脏移植,统计手术存活率和死亡原因,分析确保成功率的关键因素。结果术者通过练习,手术成功率稳定77.5%,供心冷缺血时间为(34±5)min,整个手术时间为(71±11)min。HE染色显示移植后发生了免疫反应,移植模型可靠。结论决定手术成功率的影响因素众多,其中主要因素有:合格的实验动物、供体心脏的保护、快速有效的血管缝合和术后动物护理。     

大鼠原位肝移植急性排斥反应模型的建立

目的建立大鼠原位肝移植急性排斥反应模型。方法采用改进的Limmer和Kamada的二袖套法建立大鼠肝移植模型。将大鼠分为2组：①实验组：急性排斥反应组（Wistar→SD）;②对照组：免疫耐受组（SD→SD）。结果免疫排斥组肝存活时间（7.4±1.7）d低于对照组（18.9±7.6）d,差异有统计学意义0.05（P〈0.05）。结论Wistar→SD大鼠之间的原位肝移植模型可产生中、重度的免疫排斥反应,是一种较理想的可作为研究急性排斥反应的动物模型。     

Surgical technique in the rat model of kidney transplantation

Today successful kidney transplantation procedures, techniques and immunosuppression protocols are a consequence of extensive research on animal models. During every transplantation surgery there are two crucial points for the success of the entire procedure: vascular (arterial end venous) and ureteral or ureterovesical anastomosis. Renal artery and vein of the donor kidney can be anastomosed end-to-side to the abdominal aorta and vena cava of the recipient (heterotopic transplantation), or end-to-end to the remains of renal artery and vain of the recipient (orthotopic transplantation) after nephrectomy. The ureter can be anastomosed also end-to-end or we can connect it directly to the urinary bladder (ureterocystoneostomy). The aim of this study was to elucidate which technique has better results according to: animal survival, reperfusion and perfusion of the transplanted kidney, elimination of the urine from the transplanted kidney and procedure costs. The study included 240 (120 donors and 120 recipients) male Wistar rats (3 months old; weight 250-300 g Our results are clearly showing that the end-to-end vascular anastomosis, and Paquins ureterovesical anastomosis have better results in transplanted rat kidneys survival and urine drainage compared to end-to-side vascular anastomosis and end-to-end ureteral anastomosis. Based on our experience we can conclude that described methods of end-to-end vascular anastomosis and Paquins ureterovesical anastomosis are less technically demanding and have a shorter learning curve. Therefore, we can recommend the use of described methods in kidney transplantation related researches.     

End-to-end versus end-to-side microvascular anastomosis patency in experimental venous repairs

In this experimental study, venous end-to-end and end-to-side microvascular anastomoses in similar and diameter-discrepant vessels were compared. In 50 rats, end-to-end microvascular repair of the divided epigastric vein and end-to-side repair of the epigastric vein into the femoral vein showed 5-day patency rates of 75 and 88 percent, respectively. These data are not statistically different. In 20 rats, microvascular repair of end epigastric to end femoral veins (size discrepant) and end epigastric to side femoral veins showed 5-day patency rates of 50 and 85 percent, respectively. These data are statistically different (p less than 0.05). We conclude from these experimental data that end-to-side venous repairs may be useful in lowering the anastomosis thrombosis rate seen when size-discrepant veins are repaired.     

Orthotopic Kidney Transplantation in Mice: Technique Using Cuff for Renal Vein Anastomosis

Mouse renal transplantation is a technically challenging procedure. Although the first kidney transplants in mice were performed over 34 years ago and refined some years later, the classical techniques of mouse renal transplantation required clamping both vena cava and aorta simultaneously and carry out suture anastomoses of the renal artery and vein in a heterotopic position. In our laboratory, we have successfully developed mouse orthotopic kidney transplantation for the first time, using a rapid “cuffed” renal vein technique for vessel anastomosis, wherein the donor’s renal vein was inserted through an intravenous catheter, folded back and tied. During grafting, the cuffed renal vein was directly inserted into the recipient’s renal vein without the need for the clamping vena cava and suturing of renal vein. This technique allowed for the exact transplantation of the kidney into the original position, compared to the classical technique, and has significantly shortened the clamping time due to a quicker and precise anastomosis of renal vein as described. This also allowed for a quicker recovery of the lower extremity activity, reduction in myoglobinuria with resultant kidney graft survival of 88.9%. Thus we believe that the cuffed renal vein technique simplifies microvascular anastomoses and affords important additional benefits.     

Transplantation of both kidneys from one donor rat

We have developed a technique for harvesting and transplanting two kidneys from one donor rat. Our method involves end-to-end anastomosis of the renal artery and the renal vein. Because of the short distance between the caudal vena cava and the right kidney, renal transplantation is not feasible on the right side of the recipient. The right donor kidney was therefore transplanted to the left side. Graft morphology was assessed on paraffin sections. We observed no differences between conventionally transplanted kidneys (n = 116) and right kidneys transplanted according to the method described (n = 40). In conclusion, our technique is simple, saves time and reduces the number of donor rats.     

Rotational in vitro compliance measurement of diverse anastomotic configurations: a tool for anastomotic engineering

Anastomotic configurations with a small internal diameter are prone to intimal hyperplasia which can cause occlusion within weeks or months. A link between intimal hyperplasia and inhomogenities of the elastic profile of the anastomosis has been established, making anastomotic engineering directed towards smoothing the compliance profile at the anastomotic site essential. Methods to date restrict the anastomotic compliance measurement to one plane. We present a method by which the anastomotic configurations are rotated, thereby allowing an anastomotic elastic profile assessment in multiple planes. Eight end-to-end anastomoses (ovine common carotid artery) and three end-to-side anastomoses (e-PTFE graft to ovine common carotid artery) were prepared and mounted in an artificial circulation system. Anastomotic circumferential compliance (maximal-minimal diameter/(maximal-minimal pressure.minimal diameter)) was measured by means of a laser-scan-micrometer and a Statham pressure transducer. By rotating end-to-end anastomoses, the compliance was measured in three, and in end-to-side anastomoses in four different planes. Multiplanar compliance variability in areas remote to both end-to-end and end-to-side anastomoses was approximately 9%. At the suture line the variability was approximately 22% in end-to-end anastomoses and 78% in end-to-side anastomoses. These results show that local factors result in different compliance profiles when utilizing a multiplanar technique, particularly in end-to-side anastomoses. The rotational apparatus is a tool which can be used to more accurately engineer a homogeneously compliant anastomosis, with the ultimate goal of prolonging anastomotic patency.     

大鼠颈总动脉端-侧吻合模型的建立

目的建立大鼠颈总动脉端-侧吻合模型,以期对欲进行显微血管吻合训练或相关实验的同道提供帮助。方法成年SD大鼠10只,将左侧颈总动脉远端穿过颈前肌肉群和气管之间的隧道,与右侧颈总动脉行端-侧吻合。结果成功建立大鼠颈总动脉端-侧血管吻合动物模型,手术成功率约为100%,平均吻合所需8针,平均血管吻合所需时间(35±5)min。吻合3个月后观察通畅率100%,HE染色示吻合口愈合良好。结论吻合成功的关键是提高显微操作技术水平,同时注意保护术野中小的血管和神经。此模型可以较好的应用于显微血管缝合训练。     

Preferential use of end-to-side arterial anastomoses in free flap transfers.

In our early clinical experience with free flaps, we used end-to-end arterial anastomoses and in 9 our of 24 we had complete failures--7 of which were due to early arterial thrombosis. Contrarily, in 41 consecutive free flaps with end-to-side anastomoses we have not had a single failure. At the same time that we began using the end-to-side anastomoses, we also began using the latissimus dorsi free flap as our flap of first choice, and we agree that this was probably an additional reason for our improved success rate. The use of end-to-side anastomoses has the following advantages: (1) a high success rate; (2) preservation of all existing vessels in an injured extremity; (3) greater freedom of operative planning; and (4) technical simplicity in terms of access to the vessels. For us, these advantages have made end-to-side anastomosis the technique of choice in the transfer of free flaps.