Objective To investigate the effects of Blastocyst MHC gene transfection to coronary on the survival time of mouse heart grafts and the mechanism. Methods Inbred male Balb/c mice and C57BL/6 mice were selected as donors and recipients respectively, to construct mouse cervical heart transplantation models. In the control group, the donor hearts were perfused using the 0～4 ℃ St. ThomasⅡ solution; in the cyclosporine A (CsA) group, the donor hearts were perfused as same as the control's and received intraperitoneal injection of CsA (5 rng·g-1·d-1) after surgery; in the transfection group, the donor hearts were perfused using St. Thomas Ⅱ solution with Blastocyst MHC gene plasmid; in the combined treatment group, the donor hearts were perfused using St. Thomas Ⅱ solution with Blastocyst MHC gene plasmid and received intraperitoneal injection of CsA (5 mg·g-1·d-1) after surgery. The survival time of transplanted heart allografts were observed, and their histopathological changes and the degrees of coronary intimal hyperplasia were estimated.Blastocyst MHC gene mRNA expression levels were detected by real-time fluorescence quantitative RT-PCR. Flow cytometry was applied in assessment of the levels of CD4+ CD25+ regulatory T cells (Treg) and CD3+ CD8+ T cells. Results The survival time in the CsA group, transfection group and combined treatment group was significantly longer than in the control group (P＜0.05) and that in the combined treatment group was the longest, up to (20. 50 ± 5. 61) days. On the postoperative day 1 and 3, Blastocyst MHC gene mRNA expression level in the transfection group was significantly higher than that in the control group (P＜0.05). On the postoperative day 7, the degrees of rejection and coronary intimal hyperplasia in the combined treatment group were the lightest. On the postoperative day 7 the number of Tregs in the CsA group and the combined treatment group was significantly increased as compared with that in the control group (P＜0.05), but that of CD3 + CD8+ T cells in the CsA group and the combined treatment group was less than that in the control group (P＜0.05). Conclusion Blastocyst MHC gene transfection in mouse transplanted cardiac allograft can extend its survival time through upregulation of Treg and downregulation of CD3 + CD8 + T cells in the mice. The combination of Blastocyst MHC gene and CsA may exert the synergic effects.

Objective To discuss the indication, technical points and long-term effects of endovascular embolization for non-functioning renal graft with detachable coils, and to get further evaluation of its practical value.Methods Monitored by DSA, endovascular embolization with detachable coils was performed on 11 patients with non-functioning renal graft.Results Renal arteries all had been successfully blocked in 11 cases.Good recovery without any complication was obtained.Conclusion Endovascular embolization for non-functioning renal graft with detachable coils is safe, minimally invasive and convenient, and can be used as an alternative to the resection of the renal grafts.

Objective To explore the feasibility and secure cold storage time of human arteries during sequentially cold-cryopreservation by observing the cellular metabolic activity and structure after cold storage and cryopreservation. Methods Human iliac and splenic arteries were stored for 72 h, 1 week, 2 weeks, 3 weeks and 4 weeks in UW solution at 4 ℃. After the cold storage procedure, half of the vascular allografts were examined by NBT dye method, electron and light microscope. The other vascular allografts continued to be stored by - 80 ℃ cryopreservation procedure for 4 weeks, and then the vascular allografts were examined by NBT dye method, electron and light microscope. Results There was no statistically significant difference in NBT dyeing time between the groups stored in UW solution within 2 weeks and fresh group at 4 ℃ (P ＞ 0. 05). After - 80 ℃ cryopreservation, there was also no statistically significant difference in NBT dyeing time between the groups stored by UW solution within 1 week and fresh group at 4 ℃ (P＞0. 05). Along with the extension of cold storage time, the destruction of ultrastructure was aggravated. When vascular allograft was stored over 2 weeks at 4 ℃, the destruction was more obvious. As the cold storage time prolonged, the ultrastructural destruction of vascular allografts was aggravated, especially those stored over 1 week. Conclusion The optimal time limit for arteries stored at 4 ℃ in UW solution was 2 weeks. Cryopreservation at - 80 ℃ kept the arteries satisfactory metabolic activity and organizational structure. The arteries stored within 1 week at 4 ℃ in UW solution, which restored at - 80 ℃ , could maintain satisfactory metabolic activity and organizational structure.

Objective To analyze the sensitized factors in urinemia patients who waiting for renal transplantation.Methods 2429 patients with urinemia from April 2002 to December 2008 were subjected to the detection of panel reactive antibody, and classified into 5 groups according to their clinical data:(A) no history disease group (n = 1097) who never experienced transfusion, pregnancy and transplantation; (B) Transfusion group (n = 361) who received transfusion more than 200 ml; (C) Pregnancy group (n = 481) who experienced pregnancy; (D) Transfusion+ pregnancy group (n= 294) who experienced both pregnancy and transfusion; (E) Re-transplantation group (n = 196) who experienced failed transplantation before, and waited for the second renal transplantation.Results All the males in group A were negative for PRA, and females were weakly positive for HLA Ⅱ antibody.The incidence of PRA production in group B was 15.24 % (55/361).Thirty-nine patients were positive for PRA in group C with the incidence being 8.11 % (39/481).The PRA positive rate in groups D and E was 30.61 % (90/294) and 70.92 % (139/196) respectively.PRA intensity was more than 60 % in 72 patients in group E.Conclusion Transfusion and pregnancy caused lower incidence of PRA positive rate.The incidence was much higher in transfusion + pregnancy patients than that in patients with transfusion or pregnancy alone.Graft caused the higher incidence of PRA than by transfusion and pregnancy.

Objective To evaluate the relevant causes of neurologic complications following liver transplantation.Methods 155 adult patients (131 males, 24 females) who received liver transplantation for the first time at Tongji Hospital between January 2005 and September 2009 were identified.Case notes were reviewed and demographic data, details of the liver disease, neurologic complications, MELD score and discharge information were recorded.Results Neurologic complications occurred following 36 transplants (23.2 %), The complications included mental symptoms in 15 cases (41.7 %), disorder of consciousness and action in 9 cases (25 %), and coma in 12 cases (33.3 %).Twelve percent patients with liver cancer experienced a neurologic complication, which was lower than for other transplant indications, like acute and chronic hepatic failure because of HBV infection (33.3 %, P＜0.01), inborn/metabolic disease (40 %, P＜0.05), and HCV Infection (25 %, P = 0.36).Patients who experienced a neurologic problem had significantly higher MELD score (for non-cancer patients:22.93 ± 8.21; for cancer patients:17 ± 5.4) than the other Patients (for non-cancer patients:18.33 + 8.47, P＜0.05; for cancer patients:13 ±3.4, P＜0.01).The rate of infection (36.1 %) and mortality (30.5 %) were significantly higher in patients with neurologic complications (P＜0.01).The levels of ALT, TBil, ALB, PT and the concentrations of serum sodium and chlorine had no impact on neurologic complications.Conclusion Neurologic complications are common in liver transplant recipients.These complications are related to primary disease and liver function before the operation, and increase the rate of infection and mortality.

Objective Portal hypertension and ischemia/reperfusion (I/R) have been implicated in small-for-size liver graft dysfunction. Matrix metalloproteinases-2 (MMP-2) and MMP-9 are critically involved in hepatic I/R injury. The goal of this study was to investigate the role of MMP-2 and MMP-9 in acute small-for-size graft injury. Methods 108 rats were divided into three groups:100 % (full-size), 50 % (half-size) and 25 % (quarter-size) liver transplantation groups. Blood and liver samples were collected to assess liver function, hepatic malondialdehyde (MDA) content, tissue myeloperoxidase (MPO) activity and histological changes. ELISA, real-time PCR, gelatin zymography, and immunohistochemistry were used to determine the expression pattern of MMP-2 and MMP-9 in liver grafts. Results The expression levels of MMP-9 were significantly higher in quarter-size and half-size grafts than those in full-size liver grafts 6, 12, and 24 h after reperfusioa And theelevated levels of MMP-9 were related to graft size inversely. However, MMP-2 was expressed and remained in all groups invariably. MMP-9 overexpression was accompanied by extensive liver I/R injury, as evidenced by significant increases in hepatic microscopic damage scores, MDA content,MPO activity and liver function levels. Furthermore, MMP-9 was found mainly to locate around periportal area. The presence of the active form of MMP-9 was significantly higher in small-for-size grafts, which was correlated with sinusoidal dilatation, congestion and hemorrhage. Conclusion These results support critical function of MMP-9 in acute small-for-size liver graft injury. Moreover,portal hypertension may be a crucial trigger for expression and activation of MMP-9.

Objective Islet transplantation has been an effective method for diabetes mellitus. The quality of donor pancreas is important for successful islet isolation. In this study, we evaluated expanded criteria donor usability based on the warm ischemic time, fatty pancreas and perfusion injury. Methods The marginal pancreases include those from cardiac death donor, fatty pancreas and edema pancreas from perfusion injury. Islets were isolated and purified using a modified University of Minnesota method. Islet yield and purity was determined by Dithizone (DTZ) staining and microscopic examination. Islet viability was assessed by AO/EB staining, and islet function was assessed by static glucose stimulation test. Results In the cardiac death donor group, the islet quality, viability, and in vitro function were similar when the warm ischemic time within 15 min. The quality and viability was decreased when the warm ischemic time beyond 30 min, but the function remained well. With 45 min warm ischemic time, insulin release index was decreased significantly. The islet quality, viability, and in vitro function from severe obesity group and severe edema group were decreased obviously. Conclusion Donor factors play a vital role in pancreas transplant outcomes. We concluded that pancreas severe obesity, severe edema and pancreas from cardiac donors (warm ischemic time ＞30 min) are unsuitable for islet isolation.

Objective To evaluate the value of monitoring serum cystatin C in assessment of renal function in patients with renal transplantation.Methods Serum cystatin C, creatinine (SCr), β2-microproglobin (β2-MG) and urea nitrogen (BUN) levels were determined at different time points (pre- or post-operation) in 58 renal transplant patients.Glomerular filtrated rate (GFR) was determined by using of 99mTc-DTPA at the seventh day after operation.The correlation between GFR and the four markers was analyzed.Diagnostic characteristics and ROC curve for the four markers were obtained using a GFR cut-off of 1.5 ml/s.Intra-individual coefficients of variation (CV) for cystatin C and SCr according to different time points during post-operation monitoring and the ratio (R) between CVSCr and CVcystatin C were calculated.Results Cystatin C was decreased by 48.1 % at the first day after operation, which was higher than others.The correlation coefficients between GFR and cystatin C, SCr, β2-MG, and BUN were 0.876, 0.691, 0.589, 0.516 respectively.Diagnostic characteristics for GFR and cystatin C, SCr, β2-MG, and BUN were as follows:sensitivity (91.3 %, 87.2 %, 82.6 %, 87.0 %); specificity (80.0 %, 69.2 %, 71.4 %, 42.9 %), positive predictive value (82.0 %, 73.7 %, 74.3 %, 60.4 %); positive likelihood rate (4.81, 2.83, 2.87, 1.53).The area under the curve (AUC) for GFR and cystatin C, SCr, β2-MG, and BUN was 0.914, 0.828, 0.803, and 0.765 respectively.The CVSCr was significantly lower than CVcystatin C (P＜ 0.01).R was less than 1 in most patients with cystatin C＜2 mg/L.In patients with cystatin C＞2 mg/L, R tended value 1 with increasing concentrations of cystatin C.Conclusion Cystatin C showes the best correlation to GFR, and is superior to the other markers in accurate in differentiate mild renal impairment from moderate and severe renal impairment.When renal function has minimal change, the cystatin C level has significant change.When the renal function has mild impairment, great changes in serum cystatin C indicate the unstable renal function.

Objective To explore the correlation between post-transplant glomerular filtration rate (GFR) in 1 year and long-term graft survival in renal transplant patients.Methods The clinical data of 334 patients who received their cadaveric kidney transplantations between November 1994 and October 2004 were analyzed retrospectively.According to the GFR at one year after transplant operation, normal GFR group was defined as GFR more than or equal to 1.083 ml/s, while patients whose GFR less than 1.083 ml/s were fallen into abnormal GFR group.Cockeroft-Gault (C-G) formula was used to compare the difference in the renal function between the two groups.Kaplan-Meier assay was used to compare the difference in the allograft survival between the two groups in the functional renal allograft or the non-functional renal allograft.The correlativity of GFR level at the first year and the GFR level at the 5th year was analyzed.Results The GFR level at the first year after transplantation was proportional to the graft survival time of the kidney.Five and ten years after transplantation, the renal transplantation long-term survival rate in the normal renal function groups was significantly higher than in the abnormal renal function groups (P＜0.05).As compared with the GFR level at the first year after transplantation, the changes in amplitude of GFR level at the 5th year after transplantation was (0.080 ±0.248) ml/s, and the descent had a positive correlation with GFR level at the 5th year after transplantatioa Conclusion GFR level at the first year after transplantation predicts long-term renal functioa The higher of GFR level at the first year, the higher of GFR level at the 5th year.

Objective To summarize the clinical experience of combined liver and kidney transplantation (CLKT). Methods CLKT was performed on 22 patients. The orthotopic liver transplantation (LT) was preceded with the classic fashion in 10 patients and piggyback fashion in 12 patients. The renal allograft was implanted to the iliac fossa routinely. After operation, the patients received an induction therapy with anti-CD25 monoclonal antibody or antithymocyte globulin ( ATG) and a maintenance therapy with tacrolimus (Tac), mycophenolate mofetil and prednisone. Results The CLKT was successfully performed on all 22 patients, and the graft function was restored well postoperation. During the perioperative period, an acute rejection episode of liver occurred in one patient and acute renal allograft rejection episode in 2 patients. The Tac toxicity occurred in one patient. The hemorrhage of digestive tract occurred in one recipient and the hemorrhage of peritoneal cavity in one patient. The pleural effusion occurred in 6 recipients. The pneumonia occurred in 2 cases and the peritoneal infection in one patient During a follow-up period of 6 months to 7 years 11 months, three patients died because of cytomegalovirus pneumonia in 2 patients and acute myocardial infarction in, one patient, The 1-, 3-, 5-year survival rate of recipients was 86,4 %, 81.3 %, 72.7 % respectively. Conclusion The CLKT is an effective method for treatment of patients with end-stage liver djsease and chronic renal failure.