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AIM:To investigate the effect of decitabine (Dacogen, DAC) on the proliferation and differentia-tion of K562 cells.METHODS:The K562 cells were treated with different concentrations of DAC .The colony formation ability of the cells was detected by the colony formation assay with semi-solid culture .The cell viability was detected with MTT assay.The morphologic features were observed under inverted microscope with Wright ’s staining.The changes of the cell cycle distribution and the expression of CD 11b and CD42b were analyzed with flow cytometry .The protein expression of CDK2, cyclin E1, P27, GATA-1 and PU.1 in the K562 cells was determined by Western blot .RESULTS:DAC signifi-cantly decreased the colony number of the cells and cell viability in a dose-dependent manner .The morphological changes of the cells displayed partial differentiation .After treated the K562 cells with DAC for 72 h, the cell proportion in S phase was obviously decreased , while the cell proportion in G 2/M phase was obviously increased in a dose-dependent manner . After treated the K562 cells with DAC for 7 d, the percentage of CD11b and CD14 positive cells was further elevated , and the protein expression of P27, GATA-1 and PU.1 was increased.However, the protein expression of CDK2 and cyclin E1 was decreased .CONCLUSION:DAC inhibits the proliferation and induces differentiation of the K 562 cells via regulation of cell cycle .
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Aim To observe the effects of panaxdiol saponins component ( PDS-C) extracted and isolated from Chinese ginseng herb as new Chinese patent med-icine on the promotion of hematopoiesis and the regula-tion of the immune system in treating mice models with aplastic anemia ( AA ) . Methods For preparation of immune mediated AA models, BALB/c mice were ex-posed to sublethal doses of 5. 0 Gy γ radiation, fol-lowed by transplanted lymphocytes from DBA/2 donor mice. The mice models were divided into six groups in-cluding normal control, AA model, PDS-C treated groups with lower, medium and higher dosages, cy-closporine ( CsA) as positive drug control. Both PDS-C and CsA were administered by gastrogavage for 15 days. The peripheral blood cells counts and bone mar-row pathological examination were tested, the percenta-ges of Th1/Th2/Treg cells from spleen were measured, the protein expression levels of T-bet, GATA-3 and FOXP3 transcription factors in spleen cells were detec-ted. Results Curative effect of PDS-C on treating AA mice was satisfactory. The peripheral hemoglobin, white blood cells and platelet counts in PDS-C groups with medium and higher doses were significantly higher than those in model control. Meanwhile, PDS-C ele-vated the percentages of Th2 cells and Treg cells, but decreased the percentage of Th1 cells, as well as up-regulated the GATA-3 , FOXP3 and down-regulated the T-bet protein levels. Conclusion PDS-C possesses the activities of promoting hematopoiesis obviously. It can improve marrow myelosuppression, enhance the re-covery of hematopoiestic function, and elevate the pe-ripheral blood cells counts. PDS-C also pays its immu-noregulatory efficacy though recovering from unbal-anced Th1/Th2/Treg cells in treating immune media-ted AA mice.
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[ ABSTRACT] AIM:To observe the effects of total saponins of Panax ginseng ( TSPG) on the promotion of hema-topoiesis and to explore the underlying mechanism in treating aplastic anemia ( AA) in a mouse model.METHODS:For preparation of AA model, BALB/c mice were exposed to sublethal dose of 5.0 Gyγradiation, followed by transplantation of lymphocytes from DBA/2 donor mice.The experiment was divided into 6 groups, including normal control group, AA model group, TSPG treatment groups with low, medium and high doses, and positive control group with cyclosporine A ( CsA) .Both TSPG and CsA were administered by gastrogavage.After 15 d treatment, the peripheral hemogram was test-ed, the cytokine contents in serum were measured, and bone marrow semisolid culture of colony-forming assay was conduc-ted for determining hematopoietic progenitor cells.The protein levels of extracellular signal-regulated kinase 1/2 ( ERK1/2) and its phosphorylation status in the bone marrow cells were determined.RESULTS:Curative effect of TSPG in trea-ting AA mice was satisfactory.Peripheral counts of white blood cells and platelet, and concentration of hemoglobin in TSPG groups with medium and high doses were significantly higher than those in model control.TSPG increased the colony num-bers of CFU-E, BFU-E, CFU-GM and CFU-MK derived from hematopoietic progenitor cells.Meanwhile, up-regulation of the phosphorylated ERK1/2, decreased contents of Th1 cytokines and increased contents of Th2 cytokines in the serum were observed.CONCLUSION:TSPG possess the dual efficacies on the promotion of hematopoiesis and immunoregulation in AA mice by regulating the expression of Th1/Th2/Th17 cytokines and increasing the phosphorylation of ERK1/2.
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Objective To observe the effects of whole body γ-ray radiation on hematopoiesis and cytokines related to T cell subsets in mouse,to detect the expression of transcription factors of splenic T cell subsets,and to investigate the correlation between hematopoiesis injury and abnormal immune function.Methods Totally 50 BALB/c mice were divided into radiation group and blank control group with the random number table method.The former group were given 5.5 Gy 60Co γ-ray radiation on whole body and another received sham radiation.The numbers of white blood cells and platelets of radiation group were counted at 4,8,12 and 20 d after radiation,and these numbers of blank control mice were counted only at 20 d.Hematopoietic tissue proliferation was evaluated by biopsy sections of mice femur.The contents of Th1,Th2,and Th17 in peripheral blood were detected with cytometric bead array (CBA).The expressions of T-bet/GATA-3 and RORγt/Foxp3 proteins related to the differentiation of T cell subsets in spleen tissue were measured by Western blot.Results The numbers of white blood cells and platelets of radiation group mice were reduced obviously (t WBC =18.48,15.72,9.79,3.30; t PLT =22.52,19.74,11.78,4.70,P ＜ 0.05) compared with blank control group.Biopsy sections showed that bone marrow hematopoietic cells of the radiated mice were less than those of blank group,and adipocytes became more.At 8 d,the marrow suppressions were more obvious than those at 20 d.Serum contents of Th1 cytokines IFN-γ,TNF-α and Th2 cytokines IL-4,IL-6 in the radiation group were higher than those in the blank control group at 8 and 12 d(t IFN-γ =2.93,3.36,t TNF-α =6.09,8.11,6.43,4.49,tIL-4 =4.49,3.18,t IL-6=5.11,8.67,6.67,8.55,P＜0.05).IL-17A secreted mainly by Th17 cells was also higher than the blank (t =3.68,6.24,5.32,4.06,P ＜ 0.05).Compared with the blank control group,the expression of T-bet protein increased significantly (t =5.64,2.75,3.56,4.65,P ＜ 0.05),and the expressions of GATA-3,RORγt,and Foxp3 proteins decreased at 4,8 and 12 d except the RORγt at 20 d (tRORγt =6.79,4.31,4.47,tGATA-3 =3.88,8.06,2.84,3.23,tFoxp3 =10.00,8.06,2.89,5.93,P＜ 0.05).Conclusions 5.5 Gy whole body γ-ray radiation inhibits bone marrow hematopoiesis of BALB/c mice and makes the differentiation and function of T cells to be abnormal,which may be associated with bone marrow hematopoiesis obstacle.
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Application of fresh herbs is a kind of special forms of traditional Chinese medicine. In China, there is a long and rich experience in clinical application of fresh herbs. Many studies showed that the efficacy of fresh herbs was better than that of dried herbs, but the further study about the difference of their chemical composition, effective components and the overall material basis were few. In this paper, the ideas and methods to study on material basis of the fresh herbs by comparing the difference of the fresh and dry herbs in medicine chemical composition and pharmacological activity of effective components with modern advanced separation, analysis and screening technology under the "Constituent structure theory" were proposed. It was an effectual method for studying on the reasonable development of Chinese medicine and fresh herbs resources.