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BACKGROUND:The immunomodulatory ability of bone marrow mesenchymal stem cells(BMSCs)gives it a promising future in treating graft-versus-host disease(GVHD),especially with previous success in treating patients with acute GVHD.However,there are fewer reports concerning BMSCs in treating chronic GVHD,particularly for sclerodermatous chronic graft-versus-host disease(ScGVHD).OBJECTIVE:To evaluate the efficacy and safety of treatment of BMSCs for ScGVHD,and to primarily explore the immunological mechanism of clinical efficacy.METHODS:Four ScGVHD patients at the Affiliated Hospital of Academy of Military Medicine Science,between September 2006 and August 2008,were enrolled for this trial.The median patient age was 41 years,1 female and 3 male.The patients received BMSCs infusion at a dose of(1.0～2.0)×10~7 cells every time by intrabone marrow injection from the anterosuperior iliac spine and BMSCs from the same donor for the same patient were infused more than once.Concomitant medications for ScGVHD were individualized for each patient,but all were current standard medicines and the doses were significantly tapered.RESULTS AND CONCLUTION:After BMSCs infusion,the ratio of Th1 to Th2 was dramatically overturned,with an increase of Th1 and a decrease of Th2 reaching at a new balance.Correspondingly,symptoms of all the four patients gradually improved.During the course of BMSCs treatment,the life signs and laboratory results from the recipients remained normal.By the time of this report,there has been no recurrence of leukemia in the four patients.Although this study alone cannot guarantee the application of BMSCs in ScGVHD,the results are strongly in favor of the idea that the BMSCs treatment for ScGVHD patients is therapeutically practical without any detectable side effects,which may provide a new insight into the matter of treating ScGVHD clinically,thus will greatly increase the survival rate of leukemia after allogeneic bone marrow transplantation.
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BACKGROUND:The differentiation of mesenchymal stem cells(MSCs)is strictly regulated by both genetic and epigenetic Factors .Emerging evidences have demonstrated that microRNA also plays an important role in this process.OBJECTIVE:To explore the differential expression of microRNA-125b during neuronal dliferentiation of Flk1~+ adipose-derived MSCs(AD-MSCs).MATERIALS:The fat samples were provided bv healthy female volunteers aged 15-35 years and recruited from the Plastic Surgery Hospital affiliated to the Chinese Academy of Medical Sciences.METHODS:Adult adipose tissues were obtained from healthy voluntears with age of 15-35 years .Using adherence method,Flk1~+ MSCs were obtained and the 3~(rd) passage cells were taken in the experiment.Cultured in neuronalinduction medium.these MSC were induced to differentiate towards neuronal lineage.The expression of microRNA-125b was examined at days 0,4,8 and 12.To explore its role in neuronal differentiation,we need to change its expression.RT-PCR and Taqman real-time PCR were carried out to explore the difierentially expression of microRNA-125b during neuronal differentiation of AD-MSCs.The effect of inhibitor on the expression of microRNA-125b was detected.RESULTS AND CONCLUTION:①The Flk-1~+MSCs were successfully induced into neuronal differentiation and displayed typical morphological changes 12 days after induction:Most cells retracted their cytoplasm ,fomling spherical cell body and emitted cellular protrusions.RT-PCR and immunocytochemistry studies confirmed their phenotype with expression of known neuronal cell markers including neurofilament,glial fibrillary acidic protein.②The expression of microRNA-125b was significant up-regulated during neuronal differentiation .Results of RT-PCR and Taqman real-time PCR were concordarlce with that of microRNA chip technology.③Inhibitor could down-regulate microRNA-125b.The results implied that microRNA-125b may play an important role in neuronal differentiation.
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Objective To identify the function and mechanism of MR1 in proliferation of BEL-7402 cells.Methods siRNA targeting MR1 and negative control siRNA were synthesized and transfected into BEL-7402 cells using Lipofectamine 2000.The silencing effect of MR1 siRNA was determined by semi-RT-PCR.SRB assay,colony formation assay and growth curve assay were used to investigate whether MR1 siRNA regulated cellular proliferation.Cell cycles were assessed by flow cytometry.G2 arrest reagent nocodazole was used to show the potential effect of MR1 siRNA on G1 arrest.The expression of Cyclin D1 was determined by Western blotting.Results(1)MR1 mRNA significantly decreased in BEL-7402 cells 24 h after MR1 siRNA transfection.(2)MR1 siRNA induced the down-regulation of cell growth.The expression of Cyclin D1 in MR1 siRNA tranfected BEL-7402 cells decreased significantly.(3)Flow cytometry results showed that MR1 siRNA markedly decreased G2 phase population with nocodazole treatment,and distinctly increased G1 phase population.Conclusion The gene MR1 is involved in the proliferation of BEL-7402 cells.MR1 siRNA causes inhibition of the proliferation of BEL-7402 cells.One of the mechanisms ofMR1 siRNA on the proliferation of BEL-7402 cells is the induction of G1 arrest.