Objective　To study the viability and histological change of encapsulated rat hepatocytes after being transplanted into abdominal cavity of rat. Methods　The two-step collagenase perfusing method was used to separate hepatocytes from Wistar rat liver. The separated hepatocytes were purified with Percoll density gradient centrifugation and encapsulated by the alginate-barium method. Then the purified hepatocytes were transplanted into abdominal cavity of SD rats (group 1) and the encapsulated hepatocytes were transplanted into abdominal cavity of SD rats (group 2) and Wistar rats (group 3). At different time points post-transplantation, trypan blue stain exclusion was used to determine the viability of recovered hepatocytes. The histological changes of transplanted microencapsulated hepatocytes was examined using HE stain. Results　Twenty-four h after transplantation, the viability of hepatocytes between group 1 and group 2 showed significant difference (P<0.01), but there was no significant difference between group 2 and group 3 (P>0.05). At day 4 and day 7 after transplantation, the viability of hepatocytes showed significant difference between group 1 and group 2, and group 2 and group 3 (P<0.01). At day 14 after transplantation, no significant difference was found in the viability of hepatocytes between group 2 and group 3 (P>0.05). From day 4 post-transplantation, fibrosis overgrowth was found around some microencapsules, and it was more obvious in group 2 than in group 3. Conclusions　 Microencapsulation can provide protection to transplanted hepatocytes from host immunorejection, and thus increase the viability of hepatocytes post transplantation. The existence of inadequately encapsulated microencapsule cause the fibrosis overgrowth around these capsules, resulting in ischemia and subsequent necrosis of the hepatocytes and decreasing hepatocyte viability.

Objective　To investigate the infection of hepatitis virus and spirochete in renal transplant donors and recipients to study the relationship between infection and human/kidney survival rate following renal transplantation. Methods　A total of 361 donors and 300 recipients were investigated on infection of HBV, HCV, HGV, CMV, EBV, HSV, HIV and RPR. Results　Of the 361 donors, 31 cases (8.6!%), 9 cases (2.5!%) and 2 cases (0.6!%) were found having HBV, HCV, HGV infection respectively. In the 231 recipients, the percentage of CMV, EBV, HSV, HIV and RPR carriers was 16.9!%, 11.7!%, 16.0!%, 0.4!% and 0.8!% respectively. Among the 300 grafting recipients, the infective rate of HBV, HCV and HBV plus HCV was 68.7!%, 34.7!% and 25.0!% respectively. Forty patients were randomly selected from the 300 patients, it was found that 10 (25.0!%) patients were positive for anti-HGV, 10 (25.0!%) for HGV and HBV, 5 (12.5!%) for all HGV, HBV and HCV. The percentage of CMV, EBV, HSV, HIV, RPR carriers among the 300 recipients was 49.0!%, 32.7!%, 42.0!%, 0 and 0.3!% respectively. Conclusion　Viral infectious status of the donors and recipients before operation might contribute to the occurrence of viral infection in the recipients after transplantation.

Objective　To observe the therapeutic effects and safety of angiotensin-converting enzym e (ACE)-inhibitors in the treatment of the secondary erythromatosis following ren al transplantation.Methods　Twenty-four patients with erythromatosis following renal transplantation recei ved the treatment with ACE-inhibitors. During the administration of ACE-inhibi tors, the hemoglobin, hematocrit and the side effects were observed. Results　All the patients were recovered except one who ha d to be stopped the treatment of ACE-inhibitors because of the depressing of th e blood pressure. The time of producing the effects was 7-20 days. The side effe cts included lower blood pressure accompanied by dizzy in 3 cases, anemia in 2 cases and damage to renal function in 2 cases. Conclusions　ACE-inhibitors were effective in the treatme nt of secondary erythromatosis following renal transplantation. It was important to monitor the hemogram and the renal function of the patients.

Objective　To investigate the outcome of renal transplantation in the patients with systemi c autoimmune disease. Method　The clinical data of 25 patients with autoimmune disease undergoing renal transp lantation were retrospectively analyzed. Results　 The survival rate for 1 year, 3 years, 5 years after renal transplantation in t he patients with autoimmune disease and without autoimmune disease were 88 .0 %, 80.0 %, 72.0 % and 88.9 %, 84.4 %, 77.8 % r espectively. The graft survival rate for 1 year,3 years, 5 years after renal tra nsplantation in the patients with autoimmune disease and without autoimmune dise ase were 84.0 %, 72.0 %, 60.0 % and 86.2 %, 77.0 %, 66.4 % respectively. The average intervals of dialysis pre-transplantati on b etween the patients with recurrent underlying diseases (4 patients) and with out recurrent underlying diseases (21 patients) was not difference. Among the 4 p atients with positive ANA and elevated anti-dsDNA serology pre- and post-tr ansp lant, 2 patients had recurrent underlying diseases. Conclusions　Renal transplantation should be offered to th e patients with autoimmune diseases because relapses of underlying diseases after renal transplantation seem to be rare. The patient and graft survival rate was not significantly differen t in the patients with autoimmune diseases and without autoimmune diseases.

Objective To explore the feasibility of mediating recipient lymphocyte reaction with donor dendritic cells (DCs) in renal allograft recipients. Methods Donor bone marrow monocytes (BMMCs) were isolated and cryopreserved in liquid nitrogen before kidney transplantation. At 0 day, 1month,3 month, 6 month and 9 month post-operation, CD34+ cells which were isolated from frozen BMMCs and cultured into DCs as well as the peripheral blood lymphocytes (PBLs) of donors were used as the stimulating cells to the PBLs of recipients and healthy volunteers. The number of viable DCs from frozen- and room temperature-preserved BMMCs was counted and the reactions of recipients'and healthy volunteers' lymphocytes to DCs and donor PBLs were measured. Results 6. 8 × 107BMMCs were isolated from each 10 ml of donor bone marrow on average while (4. 10 ± 0. 58) × 105CD34+ cells were isolated by magnetic active cell sorting (MACS). There was no significant difference in the isolating rate of recovered CD34+ cells at each observation point postoperatively. The percentage of viable BMMCs and CD34+ was decreased significantly at 1 month after surgery, then, decreased slowly and progressively. The decreasing rate of BMMCs was higher than CD34+. The rate of viable DCs was maintained stable (93. 2%-94. 8% ) in each group. The reactions of recipients' and healthy volunteers' lymphocytes to DCs were stronger than those to donor PBLs (P＜0. 05). The reactions of healthy volunteers' lymphocytes to DCs were maintained stable while those of recipients' were fluctuating. Conclusion Bone marrow-derived DCs are superior to PBLs in mediating long-term lymphocyte reaction after kidney transplantation due to their stable viability and stimulating ability to lymphocytes. Only once collection of a small quality of bone marrow of donors is needed to meet the demand of immune monitoring at any time after transplantation.

Objective To investigate a new strategy of bone marrow transplantation (BMT) for donor-specific tolerance induction after heart transplantation. Methods Donor bone marrow cells (BMCs)were harvested simultaneously with donor cardiac graft using modified perfusion method (PM) ,then stored in a -80 ℃ refrigerator after filtration and centrifugation. Whole BMCs (IBM-BMT) (monocytes 1.2 ×107/kg,CD34+ cells 2.38× 105/kg) in host iliac bones were injected into the bone marrow cavity 40 days after heart transplantation. Preconditoning regimens that consisted of fludarabine, antithymoctye globin and total lymphoid irradiation were performed 3 days before BMT. Tacrolimus (Tac) was administrated intravenously after BMT or orally in conjunction with mycophenolate mofetil (MMF) 3 weeks later.Cyclosporine and MMF were orally administrated 6 weeks later. Donor chimerism was detected using short tandem repeats-polymerase chain reaction in monocytes from peripheral blood at the 2nd,4th, 8th or 12th week after BMT or BMCs at the 4th, 8th or 12th week after BMT. Intramyocardium electrocardiography examination or endomyocardial biopsy was performed weekly or monthly respectively. Mixed lymphocyte reactions (MLR) were performed 3 months after BMT. Results Donor chimerism in monocytes in peripheral blood or BMCs in iliac bones measured at the 1 st,2nd and 3rd month after BMT was 26.3%, 19.1%,4.8% ,and 46.3%, 24.4%, 7.6%, respectively. After 3-month follow-up, there was no rejection confirmed by endomyocardial biopsy or intramyocardium electrocardiography. Echocardiography revealed that the diastolic and systolic function of the cardiac graft was maintained well 3 months after BMT. MLR revealed donor-specific hyporesponsiveness while immunocompetence was preserved to third-party antigens. Conclusion These findings indicate that the two-stage BMT strategy is a safe and feasible method for the induction of donor-specific tolerance via stable mixed chimerism and needs to be further confirmed after a long-term observation.

Objective To explore the perioperative application of extracorporeal membrane oxygenation (ECMO) in lung transplantation. Methods Thirty patients with primary and end-stage pulmonary disease accompanied by pulmonary hypertension were subjected to operation under the accessory of ECMO. Eighteen patients received single-lung transplantation and 12 patients bilateral sequential lung transplantation without sternal division in our hospital from November 2005 to July 2009. In 2 patients ECMO was given before operation and maintained for 19 days and 6 days respectively. In the remaining patients, ECMO pipeline was placed after anesthesia. After lung trarnsplantation,ECMO was removed after the recipients' oxygen saturation and hemodynamics were stable. Results In all recipients lung transplantation was successfully done. ECOM was removed in 27recipients after operation, and the rest 3 recipients were supported by ECMO after operation: the ECMO was removed at 36th h and 7th day after lung transplantation in two patients respectively,and another one was supported by ECMO for 5 days after operation and suffered acute kidney failure, and died of multiple organ failure 2 weeks post-transplantation. Two recipients were infected in thigh arteriovenous cut and one suffered femoral artery thrombosis, but all of them got better and discharged from hospital after treatment. Conclusion ECMO can be used for lung transplantation on patients with primary and secondary pulmonary hypertension. The complications may be associated with patients'serious condition and unstable hemodynamics. Early detection and active and effective treatment can improve patient's prognosis.

Objective To explore the influence of the lentiviral vectors-mediated mouse genetic engineering regulatory T cells (Treg) infused after allogeneic bone marrow transplantation (alloBMT) on graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect in mice.Methods Lentivirus-mediated expression of Forkhead box P3 (Foxp3) transformed CD4 + CD25- T cells from Balb/c mice into engineered Tregs in vitro. An allo-BMT model of Balb/c→C57BL/6 mice was established. The recipients were given lethal X-ray total body irradiation before transplantation.Mice were randomly assigned into five groups and each group contained 10 recipients: (1) The recipients in radiation group were injected with 0.2 ml RPMI 1640; (2) The recipients in leukemia control group were injected with 5 × 106 donor bone marrow cells and 500 mouse T-cell leukemia/lymphoma cells (EL4 cells); (3) The recipients in transplantation control group were injected with 5 × 106 donor bone marrow cells and 5 × 106 splenocytes plus 500 EL4 cells; (4) The recipients in engineering Treg group were injected with 5 × 106 donor bone marrow cells, 5 × 106 splenocytes and 500 EL4 cells plus 5 × 106 genetic engineering Treg; (5) The recipients in empty vector control group were injected with 5 × 106 donor bone marrow cells, 5 × 106 splenocytes and 500 EL4 cells plus 5 × 106 empty vector-transduced CD4+ CD25- T cells. Survival time, clinical GVHD score or histopathological analysis (skin, liver and small intestine) were observed after allo-BMT. Chimerism of bone marrow cells from recipients survived for 60 days after transplantation was measured. Results The mean survival time in radiation group, leukemia control group, transplantation control group,engineering Treg group and empty vector control group was ( 10. 3 ± 1.5), (20. 7 ± 1.9), (26. 0 ±4.3), (49. 0 ± 17. 7) and (24. 4 ± 4. 1 ) days respectively. The survival time in engineering Treg group was significantly prolonged as compared with other groups as judged by the log-rank test (P＜0. 05).Histopathological analysis in several target organs (skin, liver and small intestine) confirmed the presence of severe GVHD in transplantation control group and empty vector control group. No histological signs of GVHD or leukemia were observed in recipients in engineering Treg group and clinical GVHD scores in this group were significantly decreased as compared with transplantation control group and empty vector control group. Conclusion Co-injection of genetic engineering Treg can efficiently prevent recipients from lethal GVHD without affecting GVL activity during allo-BMT in mice.

Objective To establish and validate a standard method using flow cytometry (FCM)in human ABO blood group antibody (Ab) detection. Methods Sera samples from 52 blood donors were incubated with standardize red blood cells (RBCs), and anti-A/B antibody (IgM/IgG) was measured by indirect flow cytometry after adding secondary isotype-specific fluoresce labeled Ab. The results were compared with those by using ELISA reader. Results The changes of anti-A/B IgM measured by both methods correlated well and an overall correlation coefficiency of 0.730 for IgM was obtained by Spearman's rank testing (P＜0.05). FCM showed better specificity, sensitivity and reproducibility over ELISA reader. In blood group A samples (n = 16) or B samples (n = 16) ,the antirespectively. IgG blood group Ab could only be detected in blood group O sera. Conclusion Compared with traditional methods,FCM is a more objective method to offer accurate detection of natural ABO blood group Ab (IgM/IgG) and allowed semi-quantitative measurement of antibody levels.

Objective To investigate the effects of donor-specific regulatory T cells (Treg) transfusion on islet allograft survival. Methods Allogeneic fresh islets from Balb/c mice were transplanted to streptozotocin-induced diabetic C57 mice. The survival of islet allografts was observed. The experiment was divided into 3 groups: control group, nothing had been done to the recipients; simple islet transplantation group, the recipients received the islet transplantation only; experimental group, the recipients were given 1 ×106 Treg, then received islet transplantation. Results Blood glucose (BG) was above 16. 7 mmol/L after islet transplantation in control group; In simple islet transplantation group,BG level returned to normal level 1 to 2 days after transplantation, and hyperglycemia appeared 7 to 11 days after transplantation and maintained as the same as that before transplantation; In experimental group, BG level returned to normal level 2 days after transplantation and maintained at a low level,and at the 21st day after transplantation BG level was over 16. 7mmol/L in some recipients. Islet allograft survival in experimental group was significantly prolonged as compared with simple islet transplantation group. Conclusion Donor-specific Treg transfusion could prolong the islet allograft survival,and maybe have positive effect on tolerance induction of islet transplantation.