微囊化猪胰岛的免疫原性研究

目的 经过体外实验,明确微囊化新生猪胰岛样细胞团（ＩＣＣ）的免疫原性。方法 通过胰岛－淋巴细胞混合培养（ＭＬＩＣ）后的淋巴细胞转化试验。以游离的新生猪ＩＣＣ和空微囊为对照,比较微囊ＩＣＣ组与对照组的淋巴细胞转化值。结果 微囊ＩＣＣ组淋巴细胞转化值显著低于游离ＩＣＣ组（Ｐ〈０．０５）,但明显高于空囊组（Ｐ〈０．０５）。结论 微囊包裹后的新生猪ＩＣＣ的免疫原性比游离的新生猪ＩＣＣ的免疫原性显著降低。提     

Successful autotransplantation of isolated islets of Langerhans in the cynomolgus monkey

A recently described method for isolation of islets of Langerhans from the human pancreas has been adapted to the cynomolgus monkey pancreas. The mean yield of islets obtained from 13 monkey pancreata was 1319 islets per gram of pancreatic tissue (range 533-1800) and the usual purity of the preparation varied from 5 to 15 per cent islet tissue. Twelve cynomolgus monkeys underwent total pancreatectomy, preparation of islets from the excised pancreas and autotransplantation to either the spleen or the liver. Three animals received no transplant and became immediately diabetic, surviving 4-8 days. Four animals became normoglycaemic after intrasplenic islet transplantation, and survived 6 weeks, at which time splenectomy was performed with immediate onset of diabetes. Splenic vein insulin sampling confirmed the spleen as the source of insulin, and histological examination showed implanted islet tissue in all cases. Five animals became normoglycaemic after intrahepatic islet implantation. Three animals subsequently became diabetic at 4 and 5 months and two animals still have functioning grafts, the longest function being 9 months. These results suggest that in the cynomolgus monkey sufficient islets can be extracted from a single donor pancreas to reverse diabetes.     

Successful canine adrenal autotransplantation.

Successful canine adrenal autotransplantation has been accomplished by means of an ovarian pedicle graft. Long-term survival, normal circulating cortisol, and a positive response of circulating cortisol to ACTH stimulation was observed in four of six dogs following three operative procedures staged at 4-week intervals: (1) construction of a pedicle between the left adrenal and the left ovary, (2) excision of the contralateral adrenal (right), and (3) division of all attachments of the left adrenal gland except for the ovarian pedicle.     

Successful autotransplantation of an intra-abdominal testis by microsurgery

An intra-abdominal testis in a 12-year-old child was successfully transplanted to the scrotum by a microvascular technique. The result after 12 months is promising. Problems of ischaemia in testicular autotransplantation and technical problems in testes with a short spermatic pedicle are emphasized.     

Regulation of xenogeneic porcine pancreatic islets

The use of xenogeneic porcine pancreatic islets has been shown to be a potentially promising alternative to using human allogeneic islets to treat insulin-dependent type 1 diabetes (T1D). This article provides an overview of the existing FDA regulatory framework that would be applied to the regulation of clinical trials utilizing xenogeneic porcine pancreatic islets to treat T1D.     

微囊新生猪胰岛细胞体外培养的研究

用胶原酶消化和体外培养的方法成功地获得了大量有活性和较纯的新生猪胰岛细胞，用微囊包膜技术包裹胰岛细胞，并进行培养。由放射免疫药盒对培养液中胰岛素的含量测定，胰岛素释放试验和组织学检查等体外实验所得的数据，证实了微囊和非微囊的新生猪胰岛的活性和分泌功能的差异不显著，说明本包膜材料和一步法制囊新技术对胰岛细胞无损伤，微囊包膜后胰岛细胞的活性和分泌功能良好。     

成年猪胰岛的分离与纯化

目的　研究成年猪胰岛的分离与纯化方法。方法　胰岛的分离采用导管内连续灌注胶原酶消化法 ,纯化采用Nycodenz连续密度梯度离心。 结果　消化后每个胰腺平均可获得 (3970 0 0±185 96 3)个胰岛或 (986 0± 3 0 43)个胰岛 /克胰腺组织 ,纯化后为 (1480 0 0± 890 0 0 )个胰岛 /胰腺或(3 75 0± 493)个胰岛 /克胰腺组织 ,纯度 >95 %。体外培养示胰岛对糖刺激反应良好 ;免疫组织化学检查证实胰岛细胞存活。结论　经本法获得的胰岛具有较高的纯度及良好的功能 ,可为临床提供可靠的移植物来源。     

小鼠自体胰岛肌肉移植及疗效评价

目的探讨小鼠自体胰腺部分切除及自体胰岛的肌肉移植方法,并对术后小鼠进行疗效评价。方法选用C57BL/6小鼠,分为三组,胰腺部分切除后移植组为麻醉后切除十二指肠部沿胃大弯至脾部位的胰腺组织,并将胰岛纯化后移植至自体肌肉组织;以切除部分胰腺后未移植组和正常组作对照。结果小鼠自体胰腺部分切除和自体胰岛肌肉移植术后存活率较高,但在饲养过程中均出现持续体重下降和血糖不稳定,OGTT显示两组术后小鼠20、40 min的血糖均较正常组高,肌肉移植7 d后HE染色显示胰岛在肌肉中存活,但胰岛中心部位细胞有坏死。结论本研究建立了一种无免疫排斥下研究小鼠胰岛移植的方法,可为进一步探索改进胰岛移植研究提供参考。     

Intraportal and intrasplenic autotransplantation of pancreatic islets in the dog.

Autotransplantation of pancreatic microfragments into the liver or the spleen of totally pancreatectomized dogs is described. Both modes of transplantation resulted in restoration of normal fasting blood glucose levels. A delayed response to high glucose loads was however observed in both groups. Serum amylase levels indicated a rapid decline of exocrine activity. On the basis of postoperative levels of GOT and GPT in the serum of the dogs with intraportal transplants, permanent proteolytic or ischemic damage to the liver appeared unlikely.     

Distant processing of pancreas islets for autotransplantation following total pancreatectomy.

BACKGROUND: Small duct chronic pancreatitis is associated with intractable pain and failure to thrive, usually unresponsive to conventional management approaches. Total pancreatectomy is considered after failure of medical intervention. The major morbidity following total pancreatectomy is diabetes mellitus with its associated complications. This adverse outcome can be mitigated through autotransplantation of islets recovered from the pancreatectomy specimen. This approach has been limited historically owing to the absence of an on-site islet processing facility. We present the results from 5 pancreatectomized patients whose islets were prepared 1,500 miles away. METHODS: Five patients (4 women, 1 man, average age 42 years) who failed medical therapy and were not candidates for longitudinal pancreaticojejunostomy underwent total/completion pancreatectomy (4 total, 1 completion) for intractable symptoms from idiopathic small duct chronic pancreatitis. The resected pancreata were preserved in ViaSpan solution and were transferred to an islet processing laboratory by commercial airliner and returned. The dispersed pancreatic islet tissue was infused into a portal vein tributary through an operatively placed catheter after systemic heparinization. RESULTS: All 5 patients experienced complete relief from pancreatic pain; 2 had significant residual discomfort from underlying Crohn's disease. Three of the 5 patients had minimal or no insulin requirement after autotransplantation (median follow-up of 23 months); 1 patient continued with glycemic control difficulties related to Crohn's disease. One patient died 17 months following autotransplantation from an unrelated pneumonia. CONCLUSION: Total pancreatectomy with autologous islet transplantation can offer patients with idiopathic small duct chronic pancreatitis pain relief without the sequelae of diabetes mellitus and can be performed without an on-site islet processing facility. All patients undergoing total/ completion pancreatectomy should be considered candidates for this procedure.     

微囊化猪肝细胞培养的研究

目的　微囊包裹猪原代肝细胞并进行普通培养 ,观察肝细胞形态变化。方法　在不含小牛血清的RPMI 164 0培养基中培养 ,光镜下观察培养过程中肝细胞形态变化 ,检测不同时期培养上清中白蛋白及尿素含量。结果　微囊肝细胞可较长期存活 ,微囊肝细胞组与游离肝细胞组上清蛋白分泌在前 3d差异无显著性 (P >0 .0 5 ) ,4d后差异有显著性 (P <0 .0 5 ) ,而上清尿素合成在前 2d差异无显著性 (P >0 .0 5 ) ,3d后差异有显著性 (P <0 .0 5 )。结论　微囊材料可制备微囊化猪原代肝细胞 ,并进行体外培养 ,与游离肝细胞比较具有更好分泌合成功能。     

Successful autotransplantation of microvessel fragments into the rat heart

We examined whether microvessel fragments (Mvf), autologously transplanted with myofibroblasts (Mf) into the heart, could survive and form connections to the host's coronary microcirculation. Neither achievement has been reported before in the heart. Mvf and Mf were prepared from Sprague-Dawley rat epididymal fat pads. A mixture of Mvf (labeled with the fluorescent probe DiI-acetylated low-density lipoprotein, DiI-Ac-LDL) and Mf was injected into the superficial myocardium under general anesthesia. Rats (n = 5 in each group) were killed on postoperative days 7, 14, 21, 42 and 49, and India ink was perfused through the coronary arteries. Frozen sections of the injected area were examined under fluorescence and light microscopes. Some DiI-Ac-LDL-labeled Mvf survived in each group, and India ink was present in the lumina of microvessels coincident with DiI-Ac-LDL-labeled autotransplanted Mvf. Hematoxylin and eosin staining revealed mild inflammatory reactions followed by some fibrosis at the injection sites. These findings indicate that autotransplanted Mvf can survive for at least 49 days, and that patent microvascular anastomoses can form between them and the host's coronary microvessels. Possibly, autotransplantation of Mvf could lead to the development of a new collateral microcirculation, a phenomenon especially important in the ischemic heart.     

Histomorphometrical analysis of porcine islets of Langerhans

Xenotransplantation of pig islets of Langerhans may be an efficient method for treatment of type I diabetic patients. Considering the shortage of human pancreas donors, pigs offer readily available islet source. Unfortunately, their morphologic characteristics makes isolation of pig islets of Langerhans extremely difficult. The selection of a suitable donor may favorably affect the success of isolation. Therefore, we examined pig breeds and weights for their influence histomorphometric features of Langerhans islets. Pancreatic tissue biopsy specimens were obtained from Polish Large White pigs (n = 5), Polish Landrace pigs (n = 8), Pietrain pigs (n = 4), and marked-weight pigs of 100 kg (n = 8) or more than 200 kg (n = 10). Biopsy specimens were fixed and stored in 4% formaldehyde. Tissue sections were stained with polyclonal guinea pig anti-insulin antibody. The number of islets per 1 cm² was counted in each section. Mean diameters and areas of the islets were measured to calculate the percentage volume density. Differences were observed among the groups of pigs weighing 200 kg and 100 kg in islet mean diameter and area. The number of islets/cm² was higher among Pietrain than Polish Landrace pigs (425.45 ± 111.7 versus 315.33 ± 36.59 islet/cm², P < .05). Furthermore, Pietrain pigs showed the largest volume density (1.44%). Histological analysis revealed that Pietrain pigs of 100 kg and market-weight pigs of 200 kg were the best donors for islets of Langerhans.     

Successful ex vivo renal artery reconstruction and renal autotransplantation

Background : The increasing experience with renal allotransplantation has led to continuing development in vascular surgical techniques. These improvements have enabled complex ex vivo renal artery surgery and renal autotransplantation to be performed. The aims of the present study were to describe the results achieved with renal autotransplantation and ex vivo renal artery reconstruction (RAR) at the Newcastle Transplant Unit, John Hunter Hospital, and to review the current indications for such surgery. Methods : A retrospective review was performed of patients who required renal autotransplantation with or without RAR at John Hunter Hospital, between 1991 and 1999. Data were obtained from the Newcastle Transplant Unit and the Medical Record Department of John Hunter Hospital. Results : Two patients required ex vivo RAR and renal autotransplantation for severe fibromuscular dysplasia (FMD) complicated by stenoses and renal artery branch aneurysms. The third patient required autotransplantation for bilateral retroperitoneal fibrosis. There was one postoperative complication of pelviureteric junction obstruction that was treated successfully with a temporary ureteric stent. All patients demonstrated normal graft function and were normotensive on follow up, which ranged from 2.5 to 5 years. Conclusion : The present review confirms the long‐term benefits of ex vivo RAR and renal autotransplantation that have been demonstrated by previous studies. In transplant units experienced with this surgery it has been shown to be a successful and durable technique for the treatment of a variety of vascular, urologic and other diseases.