大鼠单纯胰腺移植手术技术

目的　探讨大鼠单纯胰腺移植(PTA)的手术技巧。方法　88 只 SD大鼠分为供、受体。根据胰外分泌系统的不同处理分为胰肠重建式(E D组,n=22)和胰膀胱重建式(B D组,n=22)行 PTA,血管吻合采用供胰腺腹主动脉袢(含脾动脉)和门静脉袢(含脾静脉)与受体腹主动脉及左肾静脉分别行端侧和端端吻合(袖套式)。于术前和术后1 d、3 d、7 d、14 d和30 d监测受体尿量、进食量、饮水量和空腹血糖。结果　平均手术时间: 供体为(33.1±11.1) min,受体为(51.7±14.7) min; 移植物均无热缺血时间,平均冷缺血时间为(46.5±17.1) min; 成功的PTA后受体术后第1天血糖即下降,第3天基本达到正常水平; 饮水量、进食量及尿量均减少,14 d后基本稳定。结论　成功的PTA可有效地恢复糖尿病大鼠内分泌功能,掌握熟练的手术技巧非常重要。     

双套管法制作大鼠胰十二指肠移植模型

目的建立并完善大鼠胰十二指肠移植模型,为临床胰腺移植提供基础研究条件。方法不阻断体循环,采用双套管将供胰所带腹主动脉和门静脉分别与受鼠的左肾动、静脉进行套管吻合,尽量减轻对全身血流动力学的影响;同时行十二指肠空肠端侧吻合引流胰液,重建胰腺外分泌途径;围手术期注意补液、保暖及抗凝。结果供体手术时间为（68.4±7.2）min,受体手术时间为（26.1±3.3）min。体外血管套管时间为（5.0±1.1）min,体内血管重建时间为（9.6±3.5）min,肠吻合时间为（7.2±2.3）min;移植成功率为91.7%。结论该移植模型不阻断体循环且操作简单,结果稳定,可用于进一步研究。     

胰腺移植免疫耐受研究进展

目的总结胰腺移植免疫耐受的研究进展。方法分析近年来有关胰腺移植免疫耐受研究进展的文献报道。结果外周性耐受及中枢性耐受是诱导免疫耐受的主要方法，其中通过供体特异性骨髓细胞输注诱导嵌合体形成是目前的研究热点。结论供体特异性骨髓细胞输注联合一种或多种外周性耐受方法有望诱导胰腺移植免疫耐受，但需进行深人研究。     

Development of Islet-Like Cell Clusters After Pancreas Transplantation in the Spontaneously Diabetic Torri Rat

Pancreas transplantation (PTx) has evolved as a clinical therapy to achieve sustained euglycemia. However, it remains unclear if naive diseased islets of the pancreas benefit from the avoidance of glucose toxicity by PTx. In the present study, using an animal model of type 2 diabetes, the Spontaneously Diabetic Torii (SDT; RT1a) rat, we syngeneically transplanted nondiabetic 10-week-old pancreaticduodenal grafts into diabetic 25-week-old recipients. In the control SDT rats that received no treatment, hyperglycemia developed with a mean onset time of 25 +/- 3.9 weeks of age. Few normal islet cells were found from 25 weeks and none at 40 weeks. However, in the PTx rats, the onset age (graft age) of diabetes was significantly prolonged (47 +/- 18.2 weeks). Moreover, we found that the beta-cell mass was significantly increased in the naive pancreases of 40-week-old PTx recipients (PTx40-naive). Interestingly, islet-like cell clusters of varying size were found close to ductal structures of PTx40-naive pancreases, suggesting that these cells are derived from ductal cells. Furthermore, pancreatic and duodenal homeobox factor-1 (PDX-1) was more clearly expressed in the nuclei of PTx40-naive pancreatic islet-like cell clusters. Our results demonstrate the development of duct-derived beta cells in the pancreas of type 2 diabetic recipients after PTx.     

一种改进的大鼠心脏异位移植模型

为移植免疫、病理和器官保存等方面的实验研究提供一种操作简便、易于观察的大鼠心脏异位移植实验动物模型。方法以Ｏｎｏ和Ｃｈｅｎ设计模型为基础，采用供心活体灌注及颈部异位移植（供心无名动脉、肺动脉分别与受体右颈总动脉和右颈外静脉作端端吻合）。结果１０例移植手术，８例移植心存活超过１月，１月存活率８０％。结论缩短手术时间、减少术中出血是手术成功的关键     

Simplified Rat Lung Transplantation by Using a Modified Cuff Technique

Mizuta's cuff technique in rat lung transplantation (LT) model has some disadvantages, such as twisting of blood vessels or bronchus and being time-consuming, which complicate procedures for anastomosis. This study was performed to investigate the advantage of using a simplified cuff technique in LT. The anastomosis time was compared in two groups. In group I, Mizuta'scuff technique was performed in 50 rat orthotopic left lung transplants. In group II, a simple modified cuff technique was performed in 48 rat orthotopic left lung transplants. The successful rate of the new technique for anastomosis was 100%. No twist of vessels or bronchus and no bleeding or air leakage were observed in group II. The anastomosis time of group II was significantly less than for group I (11.2 ± 2.1 min vs. 18.1 ± 3.6 min, mean ± SD, p <. 01). This simple modified cuff technique led to less anastomosis time and avoided potential complications induced by the cuff-tail technique. It has been verified to be a safe, simple, and reproducible technique that can provide us with a more precise assessment in the rat LT model.     

Refractory Hematuria in an Oliguric Patient After Pancreas Transplantation with Exocrine Pancreas Bladder Drainage

A 52-yr-old man presented with hematuria and clot retention. He had undergone simultaneous pancreas–kidney transplantation with exocrine pancreas bladder drainage 16 yr ago. The patient suffered from progressive transplant kidney failure with gradually decreasing urine output and needed hemodialysis every other day.     

Outcomes of Pancreas Transplantation in the United States Using Cardiac-Death Donors

Organs donated after cardiac death (DCD) are used to expand the donor pool. We analyzed the outcomes in the United States of pancreatic transplantation of organs from DCD donors performed between 1993 and 2003.
We used the OPTN/UNOS Registry to compare outcomes of primary pancreas allografts from DCD donors and donors after brain death (DBD). The primary endpoints were graft failure and patient death. A national survey regarding the use of DCD donors in pancreas transplantation was conducted among the directors of pancreas transplant centers.
Data were obtained on 47 simultaneous pancreas-kidney transplants (SPK) and 10 solitary pancreas transplants from DCD donors and on 2431 SPK and 1607 solitary pancreas transplants from DBD donors. Recipients of a SPK transplants from DCD and DBD donors had equivalent patient and graft survival rates at 1, 3 and 5 years. For recipients of SPK transplants, the wait for organs from DCD donors was significantly shorter than that for organs from DBD donors. SPK recipients of organs from DCD donors had longer hospital stays than did recipients of organs from DBD donors. With renal allografts, the incidence of delayed graft function was almost four times higher with organs from DCD donors than with organs from DBD donors.
Selective use of organs from DCD donors is safe for pancreas transplantation.     

The History of Pancreas Transplantation: Past,Present and Future

The first attempt to cure type 1 diabetes by pancreas transplantation was done at the University of Minnesota, in Minneapolis, on December 17, 1966, followed by a series of whole pancreas transplantation. Due to the lack of potent immunosuppressive drugs, rejections and infections, it was concluded that pancreas was less antigenic than the kidney which was less antigenic than the duodenum. It opened the door to a period, between the mid 70’s to mid 80’s where only segmental pancreatic grafts were used in the recipient. Numerous techniques for diverting or dealing with the pancreas juice secretion were described, none of them being satisfactory. In the late 70?s - early 80’s, three major events happened and boosted the development of pancreas transplantation: firstly the introduction of Cyclosporine A in the clinical field, secondly the organization on March 1980, of the first international meeting on Pancreas Transplantation with the first report of the International Pancreas Transplantation Registry (IPTR) and finally in 1982, the organization of the first informal so-called Spitzingsee meetings where pancreas transplantation successes but mainly failures were discussed which precluded the onset of IPITA (International Pancreas and Islet Transplantation Association), EuroSPK (European Study Group for simultaneous Pancreas and Kidney Transplantation) and EPITA (European Pancreas and Islet Transplantation Association).

During one of the Spitzingsee meetings, participants had the idea to renew the urinary drainage technique of the exocrine secretion of the pancreatic graft with segmental graft and eventually with whole pancreaticoduodenal transplant. That was clinically achieved during the mid 80’s and remained the mainstay technique during the next decade. In parallel, the Swedish group developed the whole pancreas transplantation technique with enteric diversion. It was the onset of the whole pancreas reign. The enthusiasm for the technique was rather moderated in its early phase due to the rapid development of liver transplantation and the need for sharing vascular structures between both organs, liver and pancreas. During the modern era of immunosuppression, the whole pancreas transplantation technique with enteric diversion became the gold standard for simultaneous pancreas and kidney transplantation (SPK), with portal drainage of the venous effluent of the pancreas, even for pancreas after kidney (PAK) or pancreas transplantation alone (PTA). Today, there remains room for improvement: safety of using the duodeno-duodenal anastomosis technique must be confirmed by prospective analysis while preventing ischemic reperfusion injuries, using specific drugs; that must be assessed in new trials.     

Pancreas After Islet Transplantation: A First Report of the International Pancreas Transplant Registry

Pancreas after islet (PAI) transplantation is a treatment option for patients seeking insulin independence through a whole‐organ transplant after a failed cellular transplant. This report from the International Pancreas Transplant Registry (IPTR) and the United Network for Organ Sharing (UNOS) studied PAI transplant outcomes over a 10‐year time period. Forty recipients of a failed alloislet transplant subsequently underwent pancreas transplant alone (50%), pancreas after previous kidney transplant (22.5%), or simultaneous pancreas and kidney (SPK) transplant (27.5%). Graft and patient survival rates were not statistically significantly different compared with matched primary pancreas transplants. Regardless of the recipient category, overall 1‐ and 5‐year PAI patient survival rates for all 40 cases were 97% and 83%, respectively; graft survival rates were 84% and 65%, respectively. A failed previous islet transplant had no negative impact on kidney graft survival in the SPK category: It was the same as for primary SPK transplants. According to this IPTR/UNOS analysis, a PAI transplant is a safe procedure with low recipient mortality, high graft‐function rates in both the short and long term and excellent kidney graft outcomes. Patients with a failed islet transplant should know about this alternative in their quest for insulin independence through transplantation.     

缺血预处理对大鼠移植胰腺缺血再灌注损伤的保护作用及其机制的研究

目的：探讨缺血预处理对大鼠移植胰腺冷缺血再灌注损伤和细胞凋亡的保护作用及其机制。方法：24只糖尿病SD大鼠随机分为缺血再灌注组（I/R组,n=6）和缺血预处理组（IPO组,n=18）,IPO组又根据不同方法分为3个亚组：IPO1组（再灌注30s,缺血30s1次,n=6）、IPO2组（再灌注30s,缺血30s3次,n=6）和IPO3组（再灌注30s,缺血30s6次,n=6）。24只SD大鼠为供体,I/R组和IPO组均行同种胰腺移植。另取6只SD大鼠为对照组。检测各组再灌注前、后血糖及再灌注后2h移植胰腺组织中SOD和MDA含量;TUNEL法观察移植胰腺组织细胞凋亡情况,WesternBlot检测移植胰腺组织Bax和Bcl-2蛋白表达情况。结果：1）再灌注后IPO各组相对于I/R组血糖低（P〈0.05,P〈0.01,P〈0.05）;IPO2组较IPO1组和IPO3组血糖低（P均〈0.05）;2）再灌注后IPO组较I/R组移植胰腺组织中SOD含量高（P均〈0.01）,MDA含量低（P均〈0.01）,IPO2组相对于IPO1组和IPO3组SOD含量高（P均〈0.05）、MDA含量低（P均〈0.05）。3）再灌注后IPO组较I/R组移植胰组织中AI值低（P均〈0.01）,IPO2组相对于IPO1组和IPO3组AI值低（P均〈0.05）;4）I/R组胰组织Bax蛋白高表达,Bcl-2蛋白低表达,IPO各组再灌注后移植胰组织Bax蛋白低表达,Bcl-2蛋白高表达,而IPO2组Bcl-2蛋白表达最高,Bax蛋白表达最低。结论：缺血预处理对大鼠移植胰腺的缺血再灌注损伤具有保护作用,并可以减少移植胰腺缺血再灌注后的细胞凋亡,机制与减少氧自由基、上调Bcl-2蛋白和下调Bax蛋白有关。再灌注30s缺血30s重复3次是最佳的大鼠移植胰缺血预处理诱导办法。     

A Technique for Systemic Mesenchymal Stem Cell Transplantation in Newborn Rat Pups

ABSTRACT

Mesenchymal stem cells (MSC) have the potential to aid tissue regeneration. Intravenous (IV) MSC administration is currently being assessed following tissue injury. However, few studies have been performed to establish a safe and effective method of IV MSC infusion for newborns. We have established a safe, nontraumatic and effective technique for systemic MSC transplantation in newborn rats. Yellow-fluorescent-protein (YFP)-labeled MSC were characterized using MSC markers and their differentiation potential was confirmed. Rat pups were delivered by C-section on gestational day 21. The umbilical vein (UV) was cannulated and used for IV injection of MSC or saline control, which was performed under ultrasonographic imaging. An additional control group consisted of UV MSC injection in adult mice. Mean operating time, success rate of cannulation and death rate were recorded. YFP-MSC quantification in multiple organs was performed. Mean operating time was 3.9 ± 1.1 min. The success of UV MSC injection was 92.8%. The immediate and 24 hr delayed death rate for rat pups was significantly lower than that of adult mice (p < .05). No pups receiving saline injection died. After locating the patent foramen ovale (PFO) of newborn pups by ultrasonographic imaging, extra pulse-waves and wave-shape changes were detected when MSC were injected. The number of YFP-MSC was 15.8 ± 4.1 cells per visual field (CPVF) in the lungs, 2.9 ± 1.2 CPVF in the heart, and 19.8 ± 5.0 CPVF in the intestines. We conclude that IV MSC infusion through the UV is a convenient, safe, and effective method for systemic MSC transplantation in prematurely delivered newborn rats.     

Impact of Hepatic Arterial Reconstruction on Orthotopic Liver Transplantation in the Rat

ABSTRACT

Orthotopic liver transplantation (OLT) models in rats have been investigated in many studies, but detailed information on the impact of hepatic artery (HA) reconstruction on postoperative factors remains to be investigated. HA reconstruction also requires advanced skills. The effect of the reconstruction of the HA by a hand-suture technique in rats with a whole-liver syngeneic graft was investigated. Long-term survival, histopathological assessment, immunohistological evaluation, and blood biochemistry were investigated until postoperative day (POD) 28. From the early postoperative period, significant differences between OLTs with or without HA reconstruction were found in graft parenchymal damage, induction of apoptosis, and transaminase levels, though survival curves and the coagulation profile showed no differences. In OLT without HA reconstruction, biliary proliferation was decreased at POD 5–14, and total bilirubin level was increased at PODs 10 and 14. The study indicates that HA reconstruction is required for reliable OLT in rats.     

A Rat Model of Orthotopic Kidney Transplantation Based on Nonanastomotic Technique

BackgroundRenal transplantation is an effective treatment for end-stage renal disease, which involves pathophysiologic processes such as ischemia-reperfusion injury and immune rejection. The degree of ischemia-reperfusion injury is closely related to the functional state of the transplanted kidney. At present, the allogeneic kidney transplantation model has been widely used in related research. The traditional kidney transplantation model has the disadvantages of complicated vascular anastomosis, difficulty in ureteral reconstruction. The aim of this study was to establish a rat autologous orthotopic kidney transplantation model based on non-anastomotic technique.MethodsInbred Wistar rats weighing 260 to 280 g were selected. The rats were anesthetized by intraperitoneal injections of 40 mg/kg body weight pentobarbital sodium. We exposed and freed the left kidney after laparotomy and separated the left renal artery and left renal vein, abdominal aorta, and posterior vena cava. A purse-string suture with a diameter of 1 to 2 mm was made through the tunica media of the abdominal aorta. A puncture was made through the center of the purse-string suture. The in-dwelling needle was placed in the renal artery along the blood flow direction, and was infused with constant flow of 4°C heparinized lactated ringer's solution until the kidney became pale yellow. The renal vein was ligated and the renal artery was clamped. The in-dwelling needle was removed, purse-string suture was ligated, and the kidney was stored in a self-made autologous kidney transplant cold storage bag for 4 hours. We then opened the vein and artery, removed the cold storage bag, and rewarmed with 37°C normal saline. The abdomen was then closed layer by layer.ResultsFifty-two orthotopic renal transplantations were performed, which included pre-experimental (40 operations) and experimental stages (12 operations). The success rates of the 2 stages were 75% and 91.7%, respectively. The main causes of failure were intraoperative hemorrhagic shock and postoperative infection. The operation time of orthotopic renal transplantation was 360 ± 30 minutes, including 30 ± 10 minutes for dissociation and management of kidney and blood vessels, 1 ± 0.5 minutes for warm ischemia and 240 ± 10 minutes for cold storage. Rats were sacrificed at 1 day and 7 day respectively. The rats were in good condition after operation. They could eat and drink freely. At 24 hours and 1 week after transplantation, the kidney's blood supply was good, the intestine was light or showed no adhesions, and the abdominal cavity had no ascites or peculiar smell. Hematoxylin & eosin (H&E) staining showed that there were no obvious pathologic changes in the sham group. The orthotopic kidney transplantation 1-day group showed pathologic changes of ischemia-reperfusion, such as swelling, necrosis, shedding, and cast formation of renal tubular cells. The orthotopic kidney transplantation 7-day group recovered well, with mild dilation of the renal capsule and mild dilatation of the renal tubules.ConclusionThe new model of autologous kidney transplantation is simple to use, does not require vascular anastomosis and ureteral reconstruction, and has a high success rate.     

Laparoscopic Biopsies in Pancreas Transplantation

As there is no precise laboratory test or imaging study for detection of pancreas allograft rejection, there is increasing interest in obtaining pancreas tissue for diagnosis. Pancreas allograft biopsies are most commonly performed percutaneously, transcystoscopically, or endoscopically, yet pancreas transplant surgeons often lack the skills to perform these types of biopsies. We have performed 160 laparoscopic pancreas biopsies in 95 patients. There were 146 simultaneous kidney–pancreas biopsies and 14 pancreas‐only biopsies due to pancreas alone, kidney loss, or extraperitoneal kidney. Biopsies were performed for graft dysfunction (89) or per protocol (71). In 13 cases, an additional laparoscopic procedure was performed at the same operation. The pancreas diagnostic tissue yield was 91.2%; however, the pancreas could not be visualized in eight cases (5%) and in 6 cases the tissue sample was nondiagnostic (3.8%). The kidney tissue yield was 98.6%. There were four patients with intraoperative complications requiring laparotomy (2.5%) with two additional postoperative complications. Half of all these complications were kidney related. There were no episodes of pancreatic enzyme leak and there were no graft losses related to the procedure. We conclude that laparoscopic kidney and pancreas allograft biopsies can be safely performed with very high tissue yields.     

Simulation of Reconstructive Microsurgery in Soft Embalmed Cadavers: A Teaching Module for Plastic Surgery Residents

Background  Cadaveric dissection courses—comprising flap harvesting techniques, vessel dissections, flap transfers to the defect, and microvascular anastomosis—would help residents gain confidence and master these difficult major reconstructive microsurgery procedures. Formalin embalmed bodies lack natural softness and many other features of a live body. Many soft embalming techniques have evolved to mimic live tissue and Theil technique is the most popular one among them. We explored alternate soft embalming options and started using Genelyn. Materials and Methods  Over a span of 2 years (2019–2021), we have conducted three flap dissection workshops using soft-embalmed cadavers. Six soft-embalmed and two formalin-embalmed cadavers were used. Total number of participants was 80. Results  Feedback of experience from the third course participants in the form of grades (1–5) for different criteria was obtained and evaluated. Confidence in the dissection of the various flaps and microsurgery is noticeable in all the participants. Conclusion  Based on our experience, we propose that flap dissection and microsurgery training on soft-embalmed cadavers be included as a teaching module in the plastic surgery postgraduate curriculum.     

改良单人操作大鼠原位肝移植模型的建立

目的探索改良传统手术方法单人操作大鼠原位肝移植模型的建立方法。方法 200只(100对)大鼠建立原位肝移植模型,其中80只(40对)进行正式实验,采用改良的Kamada二袖套法,再对手术中关键步骤进行改良,并对术中及术后恢复情况进行分析。结果本组80只大鼠原位肝移植手术均顺利完成。供体手术时间为(28.5±2.4)min,供肝修整时间为(10.2±1.8)min;肝上下腔静脉吻合时间为(15.3±1.9)min,门静脉吻合时间为(3.4±1.2)min,受体无肝期为(23.8±1.9)min,肝下下腔静脉吻合时间为(5.1±2.1)min,胆道重建时间为(3.1±0.9)min。整个手术过程中供、受体出血量均不足0.5 ml。40只受体大鼠,术后2 d成活率为90.0%(36/40),3只死于肝上下腔静脉缝合处出血,1只死于无肝期过长;术后7 d成活率为82.5%(33/40),3只均死于腹腔内感染。结论在单人操作建立大鼠原位肝移植模型中,通过腹主动脉和下腔静脉整块游离行腹主动脉灌注供肝、精细解剖左隔下静脉三角、术中热凝控制出血等方面的改良可减少手术并发症的发生,保证供肝质量,提高大鼠成活率。     

大鼠边缘性体积供肝肝移植模型的实验研究

目的通过建立不同体积供肝的大鼠肝移植模型,探索边缘性体积供肝大小的适宜范围,为研究小肝综合征的发病机理及防治措施提供一种易于复制的小动物模型。方法将192只大鼠随机分为全肝移植组、半肝移植组、小体积供肝移植组及超小体积供肝移植组,每组48只,分别建立全肝、半肝、小体积供肝和超小体积供肝的大鼠肝移植模型。移植术后,4组各抽取24只大鼠用于观察存活率;抽取12只大鼠于移植术前,移植术后5、15、30、45及60min测定门静脉压力;另12只大鼠于术后24h测定谷丙转氨酶（ALT）水平。结果全肝移植组的7d累积生存率为100％（24／24）,半肝移植组为87．5％（21／24）,小体积供肝移植组为37．5％（9／24）,超小体积供肝移植组均于术后48h内死亡。全肝移植组术中开放门静脉后,60min内其门静脉压力稳定;半肝移植组大鼠在开放门静脉后,其门脉压力虽有小幅升高,但仍保持相对稳定;而小体积供肝移植组和超小体积供肝移植组开放门静脉后,其门静脉压力均显著升高,15min时均达到高峰,之后该2组的门静脉压力开始回落,至45～60min时逐渐稳定。供肝的体积大小与开放门静脉后5（r=-0．942）、15（r=-0．947）、30（r=-0．900）、45（r=-0．825）和60min（r=-0．705）时的门静脉压力均呈负相关关系（P〈0．001）。肝移植术后24h,全肝移植组和半肝移植组ALT水平均低于小体积供肝移植组及超小体积供肝移植组（P〈0．05）,且小体积供肝移植组ALT水平低于超小体积供肝移植组（P〈0．05）。供肝体积的大小与大鼠移植术后的ALT水平呈负相关关系（r=-0．685,P〈0．001）。结论大鼠部分肝移植模型中,供肝与受体标准肝脏体积比（Gv／sLV）的安全界限为50％,GV／SLV为30％～35％的供肝可视为边缘性体积供肝,小于30％的供肝可视为超小体积供肝。