Investigation on Ma Peizhi’s Academic Thoughts of Surgery of Traditional Chinese Medicine / 浙江中医药大学学报

Ma Peizhi was the representative of MenHe Medicine;he learned from Ma Shengsan and Fei Boxiong,Wang Jiufeng.He was thoroughly mastered and continually blazed new trails;Ma Peizhi’s Academic Thoughts of Surgery of Traditional Chinese Medicine is still the guidance on the modern clinical experience.

Generation of hematopoietic stem/progenitor cells with property of strengthened cell mediated immunity from an embryonic stem cell line / 中国病理生理杂志

AIM: To induce lymphoid stem cells and/or T-cell precursors to diffe rentiate into functional mature T lymphocyte, and to increase the surface marker of T lymphocytes such as CD + 3, while embryonic stem(ES) cells differentiate d into hematopoietic stem/progenitor cells(HSPCs) in vitro . When they were i njec ted into lethally irradiated mice, these differentiated cells had the advantage in immune reconstitution. METHODS: Embryonic stem cells formed e mbryoid bodies(EBs) in the medium containing methycellulose, hematopoietic growt h factors(HGFs) was added to the culture system on the 6th day, thymopeptide was added at the same time. Flow cytometry were performed to detect the surface mar ker CD 34 and CD 3 of the differentiated cells. Finally the differentiated cells were injected into lethally irradiated mice, 60 days later, the incidence rate of graft versus host disease(GVHD) was taken as the mark of cell mediated immunity, PCR was performed to detect the sex determining region of the Y-chromo some(Sry) in bone marrow cells and spleen cells of the survival host female mice . RESULTS: The percentage of CD + 3 T lymphocytes was 10.52% a nd the incidence rate of GVHD was 0% on the 13th day, but they respectively rose up to 22.93% an d 100% if thymopeptide was added in the procedure of inducing ES cells to differ entiate into HSPC in vitro . CONCLUSION: The quantity of CD + 3 T lymphocytes increased in medium containing thymopeptide when ES cells differe ntiated into CD 34 + HSPC.

Primary study on inductive methods of hematopoietic stem cellfrom embryonic stem cells in vitro / 中国病理生理杂志

AIM: To explore inductive methods of hematopoietic stem cell (HSC) from embryonic stem cells (ESC) in vitro. METHODS: Using mice E14 line, the first step was the primary differentiation in which the ESC form embryoid bodies (EB) in methylcellulose-based cultures with SCF and VEGF. The second step involved the plating of cells originating from the EB into three different system of cultures containing SCF, VEGF,IL-3, IL-6 and EPO for HSC. And identifying HSC by flow cytometry analysis, colonogenic cells assay and Wright-Giemsa stain were also used. RESULTS: By two-step differentiating, it showed that HSC differentiated slowly in methylcellulose medium, percent age of CD34+/Sca-1+ cells slight increased about(31.5?4.7)% after day 14 induction. However, EBs were induced after 10 days to fast differentiate for HSC with more cells population by coculture on bone marrow stromal cells feeder. Flow cytometry analysis showed that percentages of CD34+/Sca-1+ cells might reached to (47.8?6.3)%. The more optimistic system of differentiation was bone marrow stromal cells feeder (BMSCF) in combination with supernatants of stromal cells from mice fetal liver(SSCFL), it significant supported differentiation of ESC into HSC with higher percent (53.6?7.2)%. Colonogenic cell assay and Wright-Giemsa stain confirmed that it possessed character of hematopoietic progenitors. CONCLUSION: Using methods of two-step differentiation, mice ESC were induced to differentiate into HSC by coculturing with BMSCF and SSCFL in combination of SCF,VEGF,IL-3,IL-6 and EPO.