Objective To establish the rat orthotopic liver transplantation model by characterizing the blood supply of hepatic artery with the Cuff skill and the modified arterial sleeve anastomosis, to explore the possible mechanisms of acute rejection and the express of Fractalkine (Fkn) in the early stage after hepatic allograft operation. Methods SD rats were selected as donors and Wistar rats as receptor for the rejection model of orthotopic liver transplantation. Recipient rats were divided into 2 groups randomly after operationand the drugs were given intraperitoneally once a day in each group. In the experimental group, cyclosporine A (CsA) was delivered with 3 mg/kg. In the control group, only normal saline was given with 3 ml/kg. Condition of survivals were observed. The rejection actvity index (RAI) and the expression of Fkn of liver tissue were observed after 3rd, 5th and 7th days in 5 rats. The rest of rats in each group were fed and given drug or normal saline until they were died and the mean survival time were recorded. Results There were 18 survivals in control group, and 19 in experimental group after liver transplantation. Condition of survivals in experimental group was better than that of control group. The mean survival times of experimental group(19.50±4.51 days) was significantly longer than that of control group(7.60±1.60 days), showing statistically significant difference (P＜0.05). After 3rd, 5th and 7th days of transplantation, RAI of control group were 3.80±0.35,5.90±0.87 and 7.50±1.30,respectively;RAI of experimental group were 3.10±0.21,3.90±0.41 and 4.50±0.52.Therewasstatistically significant difference in RAI between 2 groups on the 7th day after transplantation (Plt;0.01). On the 3rd,5th and 7th days after transplantation, the Fkn of control group was 8.20±0.57,21.30±3.30 and 25.70±4.91, and that of experimental group was 8.30±0.56,10.30±0.67 and 11.70±1.23. There were statistically significant differences in Fkn between 2 groups on the 5th, 7th days after transplantation (Plt;0.01). Conclusion Fkn is a participant inacute rejection after the rat orthotopic liver transplantation and can be chosen as a useful target in the diagnosis of acute rejection. CsA has immunosuppressive property in the condition of acute rejection in the rat orthotopic liver transplantation, which may be result from the decreased the level of Fkn.
ObjectiveTo study the development of methods assessing donor liver viability in liver transplantation.MethodsThe literature in the recent years on the methods of assessing donor liver viability was reviewed.ResultsFrom donor liver morphology to function,there have being developed many methods which assess donor liver viability,including:①donor liver appearance; ②intraoperative biopsies; ③donor liver microcirculation; ④portal pressure; ⑤enzymes levels in liver; ⑥lidocainemetabolizing activity; ⑦energy metabolism of donor liver; ⑧fat content in donor liver.ConclusionThere are many methods to assess the viability of donor liver. Each has its supericrity and defect respectively. Intraoperative biopsies, 31Pmagnetic resonance spectroscopy and portal pressure have more importance in clinical application.
Objective To explore the key technique of allogeneic whole pancreaticoduodenal transplantation (WPDT) in rats. MethodsWPDT model was established between Lewis rats as donors and Wistar rats (with type 1 diabetes mellitus) as recipients. End to side anastomosis was performed in abdominal aorta of donors and recipients. The portal vein of the graft was anastomosed with the recipients left renal vein by cuff technique. And side to side anastomosis was made between the graft duodenum and the host jejunum. ResultsForty-four of 50 rats were successfully performed WPDT. Amongthem, 8 rats died in postoperative 3 days, the survival time of residual 36 rats was 6-16 days, with an average of (10.45±3.30) days. The peak of death appeared on day 7-10 after operation. The typical acute rejection in pathological changes were observed on day 7. ConclusionSkilled microsurgical techniques and emphasis on details are important to establish WPDT model.
Insufficient supply of organ for allotransplantation made the study on finding new organ resources from animal progress. Pig is regarded as one of the optimal donor animals for human. The major obstacle in this field is hyperacute reaction (HAR), which is triggered after the xenogenic natural antibodies preexisting in recipient blood combine to the antigens on the surface of the endothelium and activate the complement system. alpha-Galactose residues (alpha-Gal) on the endothelial cell have been identified as the major xenoantigens. NJZ Pig has been closely breed since 1938, whose family history is clear. Tissue samples from heart, liver, kidney, pancreas, lung, small intestine, skin, spleen, thymus and lymph node were obtained and embedded in paraffin. The sections were performed the immunohistochemical staining with the sera from health volunteers (including all the blood types) as the primary antibodies as well as the biotin labeled bandeirae simplicifolia I isolectin B4 (BS I-B4), which has specific affinity to alpha-galactose. All the staining sections were compared with the tissues digested with alpha-galactosidase. There was no difference between the antigens recognized by sera of different blood types. alpha-Gal was still the major xenoantigen on the endothelial cells. There might exist non-alpha-Gal antigens on the distal convoluted tubules and collecting tubules of the kidney. There was no alpha-Gal distributing on the secreting part of pancreas, either the islet cells or the matrix cells, but surely on pancreatic duct and vessels. All the antigenity was destroyed after the enzyme digestion except that the small intestine gland still positive with the BS I-B4. alpha-Gal is the major xenogenic antigen in NJZ Pigs. There exist some unknown antigens on the distal convoluted tubules and collecting ducts of the kidney. The blood type of recipient is not the first affair to be considered in pig-to-human xenotransplantation. The specificity of BS I-B4 for the alpha-galactose needs more detail research.
Objective To study the advances in microcirculation after islets of Langerhans transplantation (ILT). Methods The literature in the recent years on the study of the relationship between ILT and microcirculation was reviewed. Results The process of angiogenesis and revascularization of the islet grafts was in progress within 1 week after transplantation, and was completed within 10-14 days after transplantation, exhibiting a microangioarchitecture similar to pancreatic islets in situ. The sequence of vascular intraislet cellular perfusion was from β cells outward to α-and δ-cell cortex, with the majority of α cells perfused before the majority of δ cells. Freely transplanted islet grafts were revascularized from the hostderived microvascular bed. The interstitial pressure in the islet transplants was markedly lower than the capillary pressure. There were clearly differences in microcirculation between syngeneic and xenogeneic islet grafts. The phenomena of microcirculation failure were observed in xenografts. The influential factors of microcirculation after ILT were ①culture temperature of isolated islets, ②cultured time and cryopreserved method of islets, ③blood glucose, ④immunosuppressive agents, ⑤angiogenesis factors. Conclusion Microvascularization of freely islet grafts is one of the essential requirements for successful engraftment, guaranteeing sufficient nutritional blood supply to the tissue and establishing blood drainage for adequate liberation of the endocrine hormones. Through the studies of the microcirculation after ILT, it is helpful to recognize the mechanism of the survival of islet grafts.
Objective To investigate the effect of transplantation of the cutaneous iliac flap on repairing bone and soft tissue defect of metatarsal. Methods From October 1999 to December 2003, the cutaneous iliac flap was designed for the repair of bone and soft tissue defect of metatarsal in 4 male patients, whose ages ranged from 26 to 47, with skin graft for the coverage of the iliac flap. The duration of injury ranged from 1 to 5 months. The length of the defect ranged from 4 to 7 cm and the defect area ranged from 5 cm×3 cm to 9 cm×5 cm. Results One week after transplantation, complete survival of iliac flap was observed in 3 cases and partial survival in 1 case (but later proved survived). Callus was seen 1 month after transplantation and Kwires were removed 4months after transplantation. Patients were followed up for 5 to 24 months. There were no ulcers. The ability of walking and bearing was satisfying as well asthe function and shape. Conclusion With satisfying restoration of function and shape, transplantation of the cutaneous iliac flap with skin graft is an alternative to treat bone and soft tissue defect of metatarsal with one stage surgery.
Objective To establish a simple, valid rat orthotopic left lung transplantation model with the improved operation technique. Methods One hundred and thirty-six male SD rats were randomly divided into donor （n = 68） and recipient （n = 68）, transplantation were performed by using the improved cuff anastomosis technique. Results Time of donor lung perfusion-picking, donor lung vessel cuff anastomosis and recipient vessel anastomosis was 13±2 min, 9±1 min, 10±1 min respectively, the operative time was 60±3 min. In 68 rats of operations, successful rate was 88%（60/68）, anastomotic stoma leak in one rat, lung congestion 3 rats, lung atelectasis 4 rats. The shortest survival time was 1 day, there were 53 rats whose survival time was longer than 12 days. The chest computed tomography showed no atelectasis and blood gas analysis manifested good respiratory function. Conclusion The improved three cuff anastomosis technique offers a simple, valid, cheap and useful method,it can establish rat orthotopic left lung transplantation model successfully.
Objective To explore the indications for liver transplantation among patients with hepatolithiasis. Methods Data from 1431 consecutive patients with hepatolithiasis who underwent surgical treatment from January 2000 to December 2006 were retrospectively collected for analysis. Surgical procedures included T-tube insertion combined with intraoperative cholangioscopic removal of intrahepatic stones, hepatectomy, cholangiojejunostomy, and liver transplantation. Results Nine hundred and sixty-one patients who had a stone located in the left or right intrahepatic duct underwent hepatectomy or T-tube insertion combined with intraoperative cholangioscopic removal of intrahepatic stones. The rate of residual stones was 7.5% (72/961). Four hundred and seventy patients who had a stone located in the bilateral intrahepatic ducts underwent surgical procedures other than liver transplantation; the rate of residual stones was 21.7% (102/470). Only 15 patients with hepatolithiasis underwent liver transplantation; they all survived. According to the degree of biliary cirrhosis, recipients were divided into 2 groups: a group with biliary decompensated cirrhosis (n=7), or group with biliary compensated cirrhosis or noncirrhosis group (n=8). There were significant differences in operative times, transfusion volumes and blood losses between 2 groups (P＜0.05). In the first group, 6 of 7 patients experienced surgical complications, and in the second, 8 recipients recovered smoothly with no complications. Health status, disability and psychological wellness of all recipients (n=15) were significantly improved in 1 year after transplantation as compared with pretransplantation (P＜0.05). Conclusion Liver transplantation is a possible method to address hepatolithiasis and secondary decompensated biliary cirrhosis or difficult to remove, diffusely distributed intrahepatic duct stones unavailable by hepatectomy, cholangiojejunostomy, and choledochoscopy.
Objective To investigate the value of monitoring cytokines in serum after human doublehand allotransplantation. Methods We detected solube interleukin-2 receptor(sIL-2R), interleukin-2(IL-2) and interleukin-6(IL-6) levels of the serum in the patient who were performed doublehand transplantation with immunosupressive from September 2000 to November 2000, and in15 healthy volunteers by using enzyme linked immunosorbent assays. Results TheIL-2 and IL-6 levels decreased quickly during inducing period （P＜0.05）, and then increased gradually. While the sIL-2R level increased. The IL-2 level was lower（P＜0.05）, the IL-6 and sIL-2R levels were slightly higher when compared with those before transplantation. There was significant difference in the IL-2level between the patient and healthy volunteers both before and after transplantation, in the IL6 level between the patient and healthy volunteers during inducing period.Conclusion IL-2, IL-6 and sIL-2R levels of serum may be useful in diagnosis of rejection, prevention of opportunistic infection, tumor and graft versus host disease.
OBJECTIVE To study the immunosuppressive effect of combined therapy with FK506 and RS-61443 in rat limb allotransplantation. METHODS: A total of 101 male SD rats were randomly divided into seven groups and used as recipients, and 101 Wistar rats were used as donors. All SD rats were performed limb allotransplantation without using immunosuppressants in control group. In experimental groups (Groups 1-6), the recipients were immunosuppressed with various dosages of FK506, RS-61443 or FK506 + RS61443, after transplantation for 5 weeks. To evaluate the results, we observed circulation of the transplanted limb, the mean rejection time, the histologic grading of skin rejection of limb grafts and the survival time of limb grafts. RESULTS: The control group showed rejection signs (edema and erythema of the skin) after a mean time of 3.36 +/- 1.15 days, and the mean survival time of the allografts was only 7.00 +/- 0.78 days. In the groups only using FK506 or RS-61443, the survival time were prolonged to varying degrees, but rejection occurred even in the period of using drug. As dosage increased, the rejection could not be prevented and the damage to liver and kidney could be induced. In the group using FK506 in combination with RS-61443, only skin and muscle of limb allografts showed slight rejection sign, function of liver and kidney was not obviously affected, the mean survival time of limb allografts was prolonged to 58.76 +/- 6.81 days. CONCLUSIONS: A combination of FK506 and RS-61443 is a more potent immunosuppressive agent than FK506 oro RS-61443 in preventing the rejection of limb allografts, and it can obviously prolong the survival time of limb allografts.