Objective To investigate the influence of indoleamine 2, 3-dioxygenase (IDO) (activity) from mesenchymal stem cells (MSCs) on the allogeneic T-lymphocytes responses.Methods MSCs were isolated and cultured from human bone marrow. Selected surface antigens of MSCs were detected by flow cytometry and their morphologic characteristics were determined by microscopy. In MSCs induced by ?-interferon (IFN-?) at different concentrations (0, 20, 50, 100, 200 U/ml), IDO mRNA expression and IDO activity were detected. MSCs Induced by IFN-? at the concentration of 200 U/ml were plated in dishes and then mixed lymphocyte reaction (MLR) cultures were set up. T- (lymphocytes) proliferation was determined by MTT assays and IDO activity was determined by high-performance liquid chromatography (HPLC).Results IFN-? could stimulate the IDO mRNA expression and IDO activity in MSCs in a dose-dependent manner. IDO enzyme activity in MSCs inhibited T-lymphocytes proliferation of MLR cultures.Conclusion MSCs could suppress T-lymphocyte responses via IDO enzyme activity in vitro.

AIM: To investigate the influence of mesenchymal stem cells (MSCs) expressing indoleamine 2, 3-dioxygenase (IDO) activity on the allogeneic T-lymphocyte responses.METHODS: MSCs were isolated and cultured from human bone marrow. Selected surface markers of MSCs were detected by flow cytometry and their morphologic characteristics were determined by microscopy. The pluripotentiality of MSCs was also studied. IDO mRNA and IDO protein expressions in MSCs induced by γ-interferon (IFN-γ) at the concentration of 2×105 U/L were detected. MSCs induced by IFN-γ at the concentration of 2×105 U/L were plated in dishes and then mixed lymphocyte reaction (MLR) cultures were set up. T-lymphocytes proliferation was determined by MTT assays and IDO activity was measured by high-performance liquid chromatography (HPLC). RESULTS: IFN-γ could stimulate IDO mRNA and IDO protein expressions in MSCs. IDO enzyme activity in induced MSCs inhibited T-lymphocyte proliferation of MLR cultures. CONCLUSION: MSCs could suppress T-lymphocyte responses via IDO enzyme activity in vitro.

Objective To investigate the diagnosis and treatment of post-transplant lymphoproliferative disease (PTLD) after hematopoietic stem cell transplantation (HSCT).Methods Clinical data of one case of PTLD,including clinical manifestations,diagnosis and treatment,were retrospectively analyzed.And the related literatures were reviewed.Results From January 2007 to November 2012,one case developed PTLD among 196 patients who were followed up after HSCT in the First Affiliated Hospital of Chongqing Medical University.The incidence of PTLD was 0.5％.Clinical manifestations of the patient were not typical,including frequent fever,multiple lymphadenopathy and multiple pulmonary nodules.The patient was diagnosed as diffuse large B-cell lymphoma (B-DLCL) by pathological examination of the swollen lymph node.After withdrawal of immunosuppressants,the temperature returned to normal,and lymphadenopathy and pulmonary nodules disappeared completely.Conclusion PTLD is a severe complication of HSCT with distinctive morphologic and clinical characteristics.The origin of this disease may be associated with Epstein-Barr virus infection and imrnunosuppressive therapy.It needs the pathologic detection to make a definite diagnosis.Several different treatment strategies have been employed,and reduction of immunosuppressive therapy may lead to regression of PTLD.

The immune mechanism of Graves' diseases (GD) and the roles of regulator T cells were investigated. In 32 patients with GD (GD group) and 20 healthy volunteers (control group), flow cytometry was used to detect the proportion of CD4+CD25+ cells, MACS to isolate CD4+ CD25+ cells,RT-PCR to assay the expression of FOXP3, and ELISA to test the level of IL-10, respectively. It was found that there was no significant change in the proportion of CD4+CD25+ T cells between GD group and control group (P＞0.05), while secretion of IL-10 and expression of FOXP3 in GD group were lower than control group (P＜0.01 and P＜0.05, respectively). In conclusion, though the proportion of regulatory T cells of peripheral blood lymphocytes in the patients with GD, the functions of them were significantly weakened, which might be a pathogenic factor in GD.

Objective:To optimize the base formula of boric acid cream,and investigate its quality. Methods:Using central composite design-response surface methodology,with the viscosity of cream as the index,the amounts of Cremophor A6,Cremophor A25,cetyl alcohol,stearyl alcohol,isooctadecyl isooctadecanoate,jojoba oil and light liquid paraffin were optimized. The appearance characteristics,particle size,viscosity and stability of the preparation and the content of boric acid were studied. Results:The optimal conditions were as follows:50 g Cremophor A6-Cremophor A25(3 ∶2),65 g cetyl alcohol-stearyl alcohol(3∶7),and 125 g isooctadecyl isooctadecanoate-jojoba oil- light liquid paraffin(5 ∶3 ∶2). According to the optimal formula,the cream showed the property of semi-fluid cream,the content of boric acid was 98. 5% of the labeling amount,and the viscosity was about 1. 64 × 104 mPa·s. The predicted value was equivalent to the measured one,and the stability of the preparation was promising. Conclusion:The central composite design-response surface methodology used in the base optimization of boric acid cream is simple with high precision and good predictability,and the quality of the preparation is stable and controllable.

AIM: To study the biology characteristics of mesenthymal stem cells(MSCs) derived from chronic myelogenous leukemia(CML) and normal adult bone marrow.METHODS: Mononuclear cells from chronic myelogenous leukemia(n=19) and normal adult(n=8) bone marrow were obtained,cultured in expanded medium with low serum concentration.Cell morphology,cell cycle,immunophenotype and in vitro differentiation capacity were investigated.The differentiations of osteocytes and adipocytes were detected by von Kossa staining and Oilred O staining.The chimeric oncogene BCR/ABL and Ph chromosome,two hall marks of CML,were detected in CML derived MSCs,normal adult MSCs,CML derived hematopoietic cells and K562 cells.RESULTS: CML and normal adult derived MSCs showed similar characteristics in cell morphology,phenotype and growth pattern.A typital fibrablast like morphology was observed.Under suitable conditions,CML and normal adult MSCs had the similar ability to differentiate into adipocytes and osteoblasts in vitro.Moreover,CML and normal adult MSCs did not express BCR/ABL gene products and Ph chromosome was not observed.CONCLUSIONS: We isolated and cultured a population of cells with characteristics of multipotent stem cells from CML bone marrow.There were similar biologic characteristics and differentiation ability between normal adult and CML bone marrow-derived MSCs.

AIM: To investigate the isolation,purification,expansion and multilineage differentiation of mesenchymal stem cells(MSCs) derived from human umbilical cord vein in vitro.METHODS: By 1% collagenase Ⅱ digestion,endothelial cells were isolated from human umbilical cord vein and cultured in IMDM medium.The morphology of the cells was observed by Wright's staining and electron microscope.Cell cycle and immunophenotype were investigated by flow cytometry.Assays of adipogenic and osteogenic differentiation were performed in vitro.von Kossa staining,Oil Red O staining and mRNA expression of osteopontin and lipoprotein lipase were studied in the induced cells.RESULTS: The cells from the cord vein displayed a fibroblast-like morphology adhering to the culture plate.FACS showed that the cells expressed several MSCs-related antigens such as CD29,CD44 and CD105,while CD13,CD31,CD45,CD34,and HLA-DR were negative.Adipocyte and osteocyte differentiation were induced successfully.CONCLUSION: The morphology,growth characteristics,immunophenotype and pluripotentiality of the MSCs from human umbilical cord vein are similar to the MSCs from bone marrow(BM).They could potentially be an excellent source of MSCs for experiments and clinics.

The immune mechanism of Graves' diseases (GD) and the roles of regulator T cells were investigated. In 32 patients with GD (GD group) and 20 healthy volunteers (control group), flow cytometry was used to detect the proportion of CD4+CD25+ cells, MACS to isolate CD4+ CD25+ cells, RT-PCR to assay the expression of FOXP3, and ELISA to test the level of IL-10, respectively. It was found that there was no significant change in the proportion of CD4+CD25+ T cells between GD group and control group (P>0.05), while secretion of IL-10 and expression of FOXP3 in GD group were lower than control group (P<0.01 and P<0.05, respectively). In conclusion, though the proportion of regulatory T cells of peripheral blood lymphocytes in the patients with GD, the functions of them were significantly weakened, which might be a pathogenic factor in GD.

Improving the special nursing ability is an important guarantee to carry out the clinical work for specialized nurse, so it is necessary to receive special hierarchical training.According to the length of employment and position, all nurse staff in the Department of Hepatobiliary Surgery ,Children's Hospital of Chongqing Medical University were classified, and the training schedule was made according to the level of personnel development to implement specialist training and operational skills training and assessment.And then examination was administrated;individualized guidance was got across to everyone according to the outcome of examination.Before and after the hierarchical training,analysis of doctors' and patients' satisfaction for nursing work, nursing theory, nursing service quality was carried out.The aim of this study is to verify the positive role of hierarchical training mode in promoting individual nursing ability and improve the level of nursing team, to provide experience and reference for nursing managers.

Objective To retrospectively analyze the influence factors during autologous peripheral blood stem cell (PBSC) mobilization and collection process in the patients with non-Hodgkin′s lymphoma (NHL).Methods Forty-eight patients with NHL in our hospital from January 2014 to August 2016 underwent PBPC mobilization and collection.The factors of age,sex,disease stage,bone marrow involvement at diagnosis,chemotherapy frequency,relapse and so on were analyzed retrospectively.Results PBSC mobilization and collection was successful in 37 cases (77.10%).The success rate was affected by the time from diagnosis to mobilization (P=0.038),chemotherapy courses (P=0.016),prior chemotherapy including fludarabine (P=0.049),relapse(P=0.033),and the levels of white blood cell (P=0.014) and platelet (P<0.01) before stem cells collection.Conclusion For the patients planning to receive autologous peripheral blood stem cell transplantation,many times of prior chemotherapy are unfavorable,mobilization and collection should be taken as early as possible,chemotherapeutic agents which possibly injure stem cells should be avoided,it is needed to simultaneously pay attention to peripheral blood WBC and platelet level for determining the collection timing.