Objective To explore the expression of the mRNA level of Fas (CD95) ligand/FasL and Fas-associated phosphatase-1/FAP-1 in acute myeloid leukemia. Methods The expression of FasL and FAP-1 were detected in 54 patients with AML and 10 normal subjects by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). β-actin used as internal reference. The changes of FasL was observed after induction chemotherapy in 16 AML patients. The expression of Fas was detected in 54 patients with AML by flow cytometry. Results The mRNA levels of FasL and in 54 patients were remarkably higher(P <0.05) than the normal controls. The rates of first complete remission(CR) were significantly higher in FasL(+) (77.78 %) than in FasL(-) cases(16.67 %) (P<0.01). The FasL level declined in 10 patients with good response to therapy (P<0.01), and high level in 6 nonresponding patients with poor response (P>0.05). The mRNA levels of FAP-1 in 54 patients was remarkly higher than that of the normal control. 8/29 cases in Fas-positive group were positive for FAP-1 mRNA expression. 19/25 cases in the Fas-refractory group didn't express FAP-1 mRNA. Conclusion The expression of FasL was high in AML. The rates of complete remission were high in FasL positive cases. The FasL level declined in patients with good response to therapy. The expression of FAP-1was partly expressed in AML. The expression of FAP-1 was less in Fas positive group.

Allograft rejection after xero organ transplantation rejection, especially acute rejection is still the major reason of failure and death. Active T cell play key roles in allograft rejection. It has been showed that the expressions of costimulatory molecules are associated with xero organ transplantation rejection. The pathways of CD28/CTLA-4 and CD40/CD40L are important costimulatory pathways that cause T cell activation.The article emphasizes on the role of CD28/CTLA-4-B7 pathway in allograft rejection.

ObjectiveTo study the effect of allo-human bone marrow mesenchymal stemcells (bMSCs) on the secretion of interleukin(IL)-1, tumor necrosis factor(TNF)-α and transforming grouth factor (TGF)-β of patients with rheumatoid arthritis (RA) in vitro. MethodsBMSCs were isolated from bone marrow of healthy volunteers and purified by density gradient centrifugation and cultured in vitro. The mononuclear cells from the peripheral blood of patients with RA and healthy controls were isolated respectively.bMSCs and mononuclear cells were co-cultured in vitro and the density of IL-1, TNF-α and TGF-β3 in the co-culture system were detected by ELISA. ANOVA and Pearson correlation were used for statistical analysis.ResultsMononuclear cells from peripheral blood of patients with RA were co-cultured with bMSCs for seven days. There were an decreased density ofIL-1[(38.4±0.5) vs(6.2±1.0) ng/L], TNF-α[(29.4±1.3) vs (4.6±1.2) ng/L]and an increased density of TGF-β[(2.6±1.0) vs (22.5±2.2) ng/L]in the co-culture system (P＜0.05). But on the other hand, for healthy volunteers there were no significant change in the density of IL1[(4.4±1.1) ng/L]and TNF-α[(5.0±1.7) ng/L]in the coculture group, as compared with the mononuclear cell group[(4.4±1.3) vs(5.3±1.7) ng/L](P＞0.05). There was an increased density of TGF-β in the coculture system[(4.8±1.4) vs(10.5±1.2) ng/L](P＜0.05). IL-1 was positively correlated to TNF-αt (r=0.896,P=0.000), TNF-β1 was nagative correlation with 1L-1 and TNF-α (r=-0.356,P=0.019; r=-0.380,P=0.000).ConclusionHuman bone marrow MSCs have modulatory effects on main cytokines of patients with RA in vitro. bMSCs could down-regulate the levels of IL-1 and TNF, but up-regulate the density of TGF-β. These immune-modulatory effects are not MHC restricted. The results of this study have provided evidence for the development of effective therapy for RA.

Objective The method of data mining and sorting analysis was used to analyze and summarize the drug experience of Professor Li Zhenhua in the treatment of chronic atrophic gastritis (CAG). Methods Professor Li’s medical records of effective diagnosis and treatment of 139 CAG patients were collected. Find-Replace method of Microsoft office word 2007 was used to count the major syndromes and main prescription of CAG. SPSS statistical software was adopted to perform entry-analysis-descriptive statistics and data analysis of and main syndrome, main formula and frequency of administration so as to obtain the commonly used drugs, commonly used prescription and drug laws of CAG. Results Professor Li Zhenhua believed that the clinical syndromes of CAG included the disharmony of liver-stomach-spleen syndrome, the damp heat of spleen-stomach syndrome, the deficiency and damp heat of spleen syndrome, the liver depression and spleen deficiency syndrome, the deficiency of spleen-stomach syndrome, the liver and stomach yin deficiency and qi stagnation syndrome, the stagnant heat of liver-stomach syndrome and the blood stasis of stomach meridian syndrome;the commonly used drugs were:bupleurum, white peony root, orange peel, licorice, poria, skullcap, ginger, fried atractylodes, golden thread, prepared pinellia, licorice, lily, stir-baking Sanxian, nutgrass galingale rhizome, heterophylly falsesatarwort root, combined spicebush root, Chinese date, tangshen, immature orange fruit, prepared rhizome pinellize without adjuvant, and oyster shell..The commonly used prescriptions were: Xiaochaihu decoction, Sini powder, Chaihu-Guizhi-Longgu-Muli decoction, Chaihu-Shugan powder, Huanglian-Wendan decoction, Banxia-Xiexin decoction, Xiaoyao powder, Xiangsha-Liujunzi decoction. Conclusion Professor Li pay attention to treat spleen and stomach disease from liver by clearing heat and removing dampness from spleen and stomach. He used the dialectical methods like invigorating qi and strengthening the spleen, regulating qi digestion, activating blood flow to eliminating blood stasis.

Objective To investigate the proliferation and the change of antigen on the cord blood CIK cells. Methods Cord blood CIK cells were generated by culture cord blood mononuclear cells in the presence of ?-IFN on day 0 and rhIL-2, antiCD3 MCAb on day 1. CIK cultures were analysed on different time point by FACS(Fluorescence activated cell sorter). The killing efficacy in tumor cell lines and primary leukemia cells were detected by MTT assay. Results After 2~3 weeks of culturing cord blood CIK cells expanded about 64.14?16 folds and CD+3 CD+56 cells which were the major cytolytic effector in CIK cells proliferated about 900 folds. Cord blood CIK cells have great cytotoxity against tumor cell lines as well as primary leukemia cells. Conclusions Cord blood MNCs can be cultured to CIK cells and the times of expansion are very high. Cord Blood CIK cells are highly efficient cytolytic effector cells.

Objective To investigate the infectious complications after nonmyeloablative allogeneic peripheral blood stem cell transplantation (NASCT). Methods Ten patients with leukemia received NASCT from HLA-identical or 1-3 antigen mismatched sibling donors with conditioning regimens of Flu/Cy+BU+ALG. Five patients were acute leukemia (AL) and all of them is in complete remission (CR1). Cyclosporine in combination with methotrexate was administered for GVHD prophylaxis. Results WBC recovered to more than 0.5 ?10 9/L during postoperative 10 day to 38 day and platelet recovered to more than 20?10 9/L during postoperative 8 day to 16 day. Bacterial infections occurred in 3 patients (30 %) and cytomegalovirus (CMV) infections in 2 patients (20 %). Varicella zoster virus (VZV) infections occurred in one patient (10 %). No fungal infections were documented. No patients died as a result of infection . Conclusion NASCT is a safe and effective therapeutic method for leukemia. It reduces acute GVHD,regimen-related toxicity and early neutropenia associated with traditional allo-HSCT.

Objective To study the distribution of allogenic bone marrow-derived mesenchymal stem cells (BM-MSCs) on joints of collagen-induced arthritis (CIA) rats and to investigate their repair effects on joint damages. Methods Five Wistar rats were used for extraction of mesenchymal stem cells and 30 adult female Wistar rats were divided into 3 groups: the CIA rats group A (n=10), CIA rats group B (n=10) and normal rats control group C (n=10). BM-MSCs of Wistar rats were isolated, cultured in vitro routinely and the fourth passages was taken for identification of specific surface antigens by flow cytometry, then the cells were labeled with 5-bromodeoxyuridine (5-BrdU) in vitro. The models of CIA rats were established. 5-BrdU labeled BM-MSCs (1.0×107 cells/kg) were imfused from through tail vein to CIA rats group A and control group C. During the first 4 weeks after BM-MSCs transplantation, changes of general condition and left hind paw swelling were examined. At the fourth week, immunohistochemical examination of 5 -BrdU and osteoprotegerin (OPG) were performed to investigate BM-MSCs aggregation around the knee joints. The contribution of BM -MSCs to repairing of joint damages was identified. Comparisons between groups were performed by t-test. Results After BM-MSCs transplantation, left hindpaw swelling of group A were relieved compared with group B (P＜0.05) and the mobility of the joints was significantly improved. At the fourth week, much more implanted cells (5-BrdU positive cells.) were detected in the damaged knee joints than those in normal knee joints. The average grey scale values on synovium of knee joints in the CIA group A (85±9) was significantly lower than that of the normal group C (110±6, P＜0.05). At the same time, OPG expression was increased in damaged knee joints. The average grey scale values on synovium of knee joints in CIA group A (54±4) was significantly lower than that of the CIA group B (77±6, P＜0.05). Conclusion The transplanted allogeneic bone marrow mesenchy-mal stem cells can migrate to sites of damaged tissue in arthritis. They can prevent tissue damage and repair the damaged joints tissue by increasing OPG expression. This study has provided some evidence for developing effective therapy for rheumatoid arthritis.

Objective To study the effect of allo-human bone marrow mesenchymal stem cells (BMSCs) on T and B cells from patients with Rheumatoid arthritis (RA) in vitro. Methods BMSCs were isolated from bone marrow samples of healthy volunteers and purified by density gradient centrifugation and cultured in vitro. Peripheral lymphocytes were isolated from patients with RA.Then, BMSCs and lymphpcutes were co-cultured. The modulatory effect of BMSCs on proliferation, activation and maturation of T and B lymphocytes of RA patients stimulated by PHA and SAC respectively was observed. The cell generation cycle and the degree of apoptosis were assessed by flow cytometry with PI/ Annexin V. After co-cultured with or without BMSCs for 72 hours, T cells were harvested, then they were labeled with anti-CD3, anti-CD4, anti-CD8, anti-CD25 antibodies and analyzed by flow cytometry. The density of IgG in the co-culture system was detected by ELISA. Results T and B cells proliferation was significantly suppressed when co-cuhured with bMSCs but did not induce T cell apoptosis. There was a significant decrease in the ratio of CD4+ CD3+ T cells in the co-cuhure group (34±6), as compared with the control group (44±7) (P<0.05). There was a decrease in CD25+ T cells and increase of CD4+ CD25+ cells in BMSCs co-cultured group (P<0.05). IgG was in creased in the cocuhure system. Conclusion Human BMSCs inhibit T and B cell activation and proliferation in patients with RA in vitro. And these immunomodulatory effects are not MHC restricted. The results of this study have provided evidence for the fact that BMSCs has the potential to be an effective treatment for RA.

Objective This study was designed to explore the influence about apoptosis of STI571 on NB4.Methods After NB4 cells were treated with STI571 in different concentration(0.5,1.0,5.0umol/L)at indicated time(24h,48h,72h)(1)morphological observation:To determine cell morphology by light microscope.(2)MTT assay:Inhibition of proliferation was measured with a colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-dipheny tetrazolium bromide(MYa3 assay.(3)Flow cytometric analysis:The expression of survivin and smac were measured on NB4 by FCM.Results Morphological observation showed characteristic apoptosis changes.MTT assay showed that STI at (0.5,1.0,5.0 μmol/L) range for 24 h,48 h and 72 h can markedly inhibit the proliferation of NB4 cells in a dose-dependent manner as well as a time-dependent manner(P＜0.05).FCM showed the expression of survivin decrease and smac increase.Conclusion STI571 In Vitro inhibits proliferation of NB4.

Objective To analyze the relationship between the expression of protection of telomeres 1 (POT1) and the pathogenesis of acute myeloid leukemia (AML). Methods 62 patients with de novo AML (case group) and 10 patients with iron deficiency anemia (control group) were enrolled in this study. The quantitative real-time polymerase chain reaction (PCR) and Western blot were used to detect the expression of POT1 in AML patients. Results There were 62 de novo AML patients, including 2 cases M1, 14 cases M2, 12 cases M3, 14 cases M4, 17 cases M5, 2 cases M6 and 1 case AML without classification. According to the risk stratification, high risk group (24 cases), medium risk group (22 cases) and low risk group (16 cases) were divided. Compared with that in the controls, POT1 expression levels in patients with AML were significantly decreased both in mRNA and protein level (P< 0.05). The relative expression levels of POT1 mRNA and protein in patients with M2, M4 and M5 were significantly lower than those in the controls (P< 0.05). The expression levels of POT1 in high risk group, medium risk group and low risk group were significantly decreased than those in the controls (P<0.05). Compared with that in the controls, The relative POT1 mRNA expression was significantly decreased in M3 patients (P< 0.05), but not in protein level. POT1 protein expression was showed both in the cytoplasm and nucleus. There was no significant difference of the expression of POT1 protein between cytoplasm and nucleus (P> 0.05). Conclusions POT1 may be involved in the pathogenesis of AML. POT1 protein expresses in both cytoplasm and nucleus, and the regulatory mechanism may be related to the telomere length.