Objective To establish a new technique isolating pancreatic islet of langerhans and evaluate the clinical efficacy and safety of adult islet transplantation.Methods Pancreases were stored using the "2-layer method" of the oxygenated perfluorochemical (PFC) and UW solution. The Pancreases were digested by Liberase collagenase enzyme and purified using continuous gradients of Ficoll-diatrizoic acid on a refrigerated COBE 2991 centrifuge to separate the islets. Cultured islets were infused by surgical approach to the liver via portal vasculature. Clinical metabolic data such as blood glucose, dose of insulin, C-peptide, HbA1c, liver function and renal function, was determined and compared with the pre-transplant data.Results Islets of langerhans were isolated successfully in 42 pancreases. The average of islet yield was 285000 islet equivalents(IEQ). Islet purity and viability were 95.7%, 93.2%, respectively. The stimulation index(SI) as assessing function of human islet was 2.43 and negative-etiology in vivo. Twenty clinical islet transplant infusions have been carried out in 11 subjects with type 1 diabetes mellitus(T1DM). The average islet mass for infusion was 11200 IEQ/kg. The treatment strategies for islet transplantation was glucocorticoid-free immunosuppressive regimen. During 7 months to 4 years follow-up, 7 recipients had insulin independence, the dosage of insulin decreased by 60% in 4 patients after islet transplantation. The level of blood glucose and HbA1c, liver and renal function were normalized throughout follow-up period. All patients had C-peptide positive after islet transplantation. No adverse effects and complications related to islet infusion procedure.Conclusions New technique has proved to be suitable for isolating pancreatic islet of langerhans. Adult islet transplantation can be used as an effective and safe way for treating T1DM.

Objective To discuss the clinical features of primary epididymal rhabdomyosarcoma and review the diagnostic and therapeutic advances. Methods One 16-yesr-old male presented with right scro-turn swelling mass for 4 months. Physical examination revealed an ovoid and solid mass of 4.5×3.5×3.0 cn in right scrotum. The mass was hard and its surface was smooth without tenderness. Transillumination test was negative. There was no palpable inguinal lymph node. Pre-operative diagnosis was inflammatory nodule of right epididymis. Results Right epididymectomy was performed under caudal anesthesia une-ventfully. The pathological study showed that the neoplasm was composed of small round shape ceils with small and anachromasis nucleus. The immunohistochemical study showed that these cells were positive for myoglobin, aetin and desmin, but negative for smooth muscle actin. Electron microscopy showed that paral-leling thin filaments were found with no asrcomere-like structure in the intracytoplasm. The pathologic diag-nosis was right epydidymal rhabdomyosarcoma. The patient refused further therapy. A mass was found with rapid growth accompanied with pain in the right scrotum 2 months after the first operation and was clinically diagnosed as tumor recurrence. This patient had then undergone right radical orchiectomy. Frozen section analysis revealed negative surgical margin. The pathologic diagnosis was embryonal rhabdomyosarcoma re-currence. Patient underwent subsequent chemotherapy with ifosfamide, etoposide and vincristine. After one year follow-up, he was generally well without evidence of recurrence. Conculsions Primary embryonal rhabdomyosarcoma is a rare tumor with aggressive growth patterns. The diagnosis depends on the pathologi-cal examination because the clinical presentation is not specific. Combination therapy with radical orchiecto-my, chemotherapy and radiotherapy provides patients an excellent long-term prognosis.

Natural killer T (NKT) cells,a subset of lymphocytes that bridge innate and adaptive immune systems,involve in processes of infection immunity,tumor immunity,transplantation immunity and autoimmunity.A significant progress has been made in the mechanisms of origin,selection,differentiation and maturation of NKT cells.However,some viewpoints are still controversial,and need to be further intensively investigated.The potential therapeutic applications of functional NKT cells have been suggested in the prevention and the treatment of various diseases.

Objective To evaluate the values of cell-mediated immune function in elderly renal allograft recipients.Method The levels of immuknowTM ATP was sequentially monitored by means of Cylex immuknowTM assay in 52 elderly renal allograft recipients including 11 with infection and 8 with acute rejection.Results No statistically significant difference was found between stable allograt function and uremia (P＞0.05).The levels of immuknowTM ATP during infection was significantly lower than those with stable allograft function with acute rejection (P ＜ 0.01).The levels of immuknowTM ATP during acute rejection was significantly higher than those with stable allograft function with infection (P＜0.01).Conclusion Sequential monitoring of immuknowTM ATP is helpful for elderly renal allograft recipients in individualized immunosuppression therapy.Cylex immuknowTM assay can be used as a potent tool for assessment of high risk in infection and rejection.

Objective To evaluate the life quality in front and at the back of the cure of the floating population tuberculosis patients.Methods The SF-36 scale was applied to the new smear-positive pulmonary tuberculosis patient in floating population using face to face interview in Shenzhen 6 districts.The contrast was tuberculosis patient in the population of the household register,the people stayed temporarily in the same term.Results The score of general health(43.36 ± 15.67),vitality(57.96 ± 16.54) and mental health(61.46 ± 14.26) in the floating population was lower than the appraisal result with the population of the household register(49.28 ± 17.85,66.38 ± 17.03 and 66.70 ± 15.12),the people stayed temporarily(45.00 ± 17.22,60.99 ± 16.59 and 63.13 ±13.71 ) (P ＜ 0.05 ).It was still lower than the other two groups after 2 (3) months and 6 (7) months (P ＜ 0.05 ).But the score of social functioning in floating population ( 88.92 ± 14.88 ) was higher than the other two groups (82.62 ± 14.44,68.02 ± 27.30).The change rate of general health ( (76.05 ± 132.82) ％,(69.32 ± 111.98 ) ％and (48.36 ± 101.31 ) ％ ) and social functioning scores( (46.49 ± 85.18 ) ％,( 26.33 ± 50.25 ) ％ and ( 51.09±73.44)％ ) were significant difference in the three groups(x2 =12.088,P=0.002;x2 =11.611,P=0.003).The change rate of the general health score in floating population was higher than the household register population and people stayed temporarily.Conclusion It is necessary to implement the management in whole course to the floating population tuberculosis patients.

Alemtuzumab (Campath) was successfully injected in 21 kidney transplant patients,7 islet transplant patients and 1 simultaneous kidney and islet transplant patient for either prevention or treatment of graft rejection.Prophylactic administration was successfully completed in all patients without discontinuation.Adverse events were not observed in 11 patients (38%),but hypertension in 18 patients (62%),shivering in 3 patients (10.3%),high fever in 3 patients (10.3%),and bronchospasm in 1 patient (3%),respectively.All complications alleviated after proper therapy.During the prophylactic administration of alemtuzumab,strict,timely and proper ward-management was needed.Care for lung,perineum,skin,diet and psychological nursing were necessary.Neither graft acute rejection nor graft chronic rejection episode occurred in all patients during 6 months to 2 years follow-up.Therefore,long term effects of Alamtuzumab and consequences of lymphocytopenia need further observation.

Objective To determine the functions of CCR7+CD8+CD45RO+ T cells on CD4+T cells in systemic lupus erythematosu(SLE).Methods The expres sion of cytokines in CD4+T cells was measured by flow cytometry,real-time qua ntitative reverse transcription polymerase chain reaction and Northern blotting.A chemotaxis assay was used to detect their functions.Results In the case of active SLE,CCR7+CD8+CD45RO+T cells could induce CD4+T cells to express high le vels of Th2-cytokine (IL-4),and low levels of Tr1-cytokines (IL-10 and TGF-?),than those in normal controls and inactive SLE (P

The effects of PEG10 on hydrogen peroxide (H(2)O(2))-induced apoptosis in human normal liver cell line L0(2) were investigated. The PEG10 gene was transfected into L0(2) cells by lipofectamine, the positive clone was screened by G418 and defined as L0(2)/PEG10, while the cell transfected with empty expression vector (pEGFP-N1) was defined as L0(2)/vector. L0(2)/vector and parental L0(2) cells served as control. RT-PCR and Western blotting were employed to detect the expression of target genes. H(2)O(2) (50-400 mmol/L) was administered to induce the apoptosis of L0(2) cells. Cells viability was measured by MTT and the morphological changes of apoptotic cells were determined by fluorescence microscopy using hoechst33342 nuclei staining. DNA fragmentation was observed by agarose gel electrophoresis. PEG10 mRNA and protein levels in L0(2)/PEG10 cells were significantly increased as compared with those in the control cells. After treatment with 400 mmol/L H(2)O(2) for 24 h, the cellular growth inhibition rate of L0(2)/PEG10 cells was significantly lower (58.2%) than that of L0(2) (92.5%) and L0(2)/vector (88%). Distinct morphological changes characteristic of cell apoptosis such as karyopyknosis and conglomeration were not observed in L0(2)/PEG10. Ladder-like DNA fragmentation in a dose-dependent manner was observed in both L0(2) and L0(2)/vector cell lines, but not in L0(2)/PEG10. PEG10 over-expression significantly inhibited cytotoxicity induced by H2O2 on human normal liver cell line L0(2) by antagonizing H(2)O(2)-induced apoptosis.

BACKGROUND:Bone mesenchymal stem cells have immunological regulation function both in vitro and in vivo, while the effect of bone marrow mesenchymal stem cells on CD4+T cellimmune function in patients receiving kidney transplantation remains unclear. OBJECTIVE:To explore the monitoring significance of CD4+T-cellimmune function by ImmuKnow assay and to determine the effect of induction therapy with bone marrow mesenchymal stem cells on cellimmune function in patients receiving kidney transplantation. METHODS:From January 2011 to June 2013, 24 patients receiving al ograft renal transplantation with autologous bone marrow mesenchymal stem cells were included and another 48 patients receiving al ograft renal transplantation and Simulect induction therapy with various matched preoperative characters served as controls. In both groups, adenosine triphosphate levels in CD4+T cells in the peripheral blood were determined by the ImmuKnow assay preoperatively and at 14, 30, 60, 90, 180 days postoperatively, as wel as during acute rejection and infection episodes. RESULTS AND CONCLUSION:During the 180 days postoperatively, fewer patients in the bone marrow mesenchymal stem cellgroup had acute rejection and injection than the Simulect group, but no significant differences were observed. Postoperative adenosine triphosphate levels in CD4+T cells were significantly lower than those determined preoperatively in both groups (P<0.05), while no significant differences were observed between the two groups. A total of 12 patients in the bone marrow mesenchymal stem cellgroup and 26 patients in the Simulect group had infection episodes, and the adenosine triphosphate levels in CD4+T cells during the infection episodes were lower than clinical stable patients in both groups (P<0.01). For patients receiving renal transplantation, induction therapy with bone marrow mesenchymal stem cells can effectively decrease the cellimmune function, which can be reflected by the adenosine triphosphate levels in CD4+T cells in the peripheral blood determined by the ImmuKnow assay.

Objective To analyze the clinical application of donor specific antibodies (DSAs) detected by a single antigen Luminex virtual crossmatch,and to discuss the treatment of DSA and the impact of DSA on renal function.Methods Serum from living-relative renal recipients before and after transplantation was investigated using a Luminex single antigen assay.The relation between DSA and renal acute rejection as well as renal function was analyzed.Results A total of 30 patients and 173 serum samples were tested,including 47 serum samples before transplantation,and 126 after transplantation.DSA was positive in one patient before transplantation,and 8 patients after transplantation.Three of the patients positive for DSA were treated by Bortezomib,3 by addition of MMF,2 by addition of CNI,1 by addition of Sirolimus.The MFI of DSA in one of the patients treated by Bortezomib was decreased to below 1000,while that in the other two decreased by more than 50 ％.The renal eGFR at the time with and without DSA was (1.50 ± 0.59) and (1.23 ± 0.38)ml/s respectively (P＜0.05).Conclusion Dynamic monitoring of single bead antigen antibody DSA conduces to direct the adjustment of immunosuppressant.The appearance of DSA contributes to the declination of renal function.Application of Bortezomib decreased the MFI of DSA.