Objective:To evaluate the expression of basic fibroblast growth factor (bFGF) in fetal liver stromal cell(FLSC) during hypoxia so as to provide evidences for hematopoietic stromal cell therapy of brain ischemia. Methods: FLSCs from 14.5-day-old mouse fetuses were cultured and passaged. Subconfluent cells of the fifth passage were cultured in hypoxia condition (95% N_(2) + 5% CO_(2) ). bFGF mRNA and bFGF protein were detected at 2,4,8,16 and 32 h of culture by semi-quantitative RT-PCR and Western blot, respectively. Results: bFGF mRNA and bFGF protein were very low in normally cultured cells, but increased significantly under the condition of hypoxia. Expression of bFGF mRNA obviously increased at 2 h of hypoxia culture and reached peak at 16 h, with the peak-value being 5.52 folds that of the control (-[32.86?9.57-]% vs -[5.94?2.65-]%,P

Objective:To evaluate the expression of transforming growth factor-?1 (TGF-?1) in fetal liver stromal cells during hypoxia,in an attempt to provide evidences for cell therapy of brain ischemia. Methods: Cells from 14.5-day-old mouse fetuses were cultured and passaged. Subconfluent cells of the fifth passage were cultured in hypoxia condition (95% N2 + 5% CO2 ). TGF-?1 mRNA and TGF-?1 were detected by semi-quantitative RT-PCR and Western blot,respectively. Results: TGF-?1 mRNA and TGF-?1 levels,which are very low in normally cultured cells, increased significantly in hypoxia-cultured cells. Expression of TGF-?1 mRNA increased from (5.47?2.76)% to (10.38?3.65)% at 4 h of hypoxia(P

AIM: To investigate the variation and distribution of abnormaly methylated p15 INK4B in myelodysplastic syndrome (MDS) and its subgroups. METHODS: The abnormal methylation of p15 INK4B in 32 cases with MDS was studied using methylation-specific PCR (MSP). RESULTS: The positive rate of abnormal methylation of p15 INK4B was about 50% in MDS. The ratios in subtypes were: 0% (0/6) in RA,20% (1/5) in RA-S,57.1% (4/7) in RAEB,74.1% (5/7) in CMML,85.7% (6/7) in RAEB-t, respectively.It was worth noticing that 4 cases represented abnormal methylation of p15INK4B during their transformation and progression into AML. CONCLUSION: The abnormal methylation in p15 INK4B might be one of the causes of MDS, which was related to pathologial process of MDS.Every subtype was not solitary classification completely. Abnormal methylation of p15 INK4B was apt to occur accompanying the progression and transformation of the subtypes.

AIM: To study the effect of IL-12 on T lymphocytes apoptosis, the expression of Fas/FasL and TNFR/TNF?. METHODS: Terminal dUTP nick end labeling(TUNEL) and Annexin V assay were used. Anti-TNFR were labeled with FITC, anti-CD95 was labeled with PE and Anti-FasL with biotin. Three kinds of T cells (HTB176,TIB152 and human normal T cells) were analysed through flow cytometry. RESULTS: At 1st hour after being treated with IL-12, the expression of FasL protein and FasL mRNA in HTB176 and TIB152 began to increase and reached peak value in 24 hours. In the normal T cells, FasL just began to increase in 1 hour and maintained stability in 6, 12 and 24 hours through the later experiment period. All three kinds of T cells displayed no change in the expression of CD95 and TNFR/TNF? under the stimulation of IL-12. CONCLUSION: Expression of such apoptosis regulating factors were different in the apoptosis of T cells induced by IL-12.

[ABSTRACT]AIM:ToinvestigatetheeffectofmiR-155-specificsiRNAaloneorincombinationwithcytosinear-abinoside (Ara-C) on the growth and apoptosis of Burkitt lymphoma Raji cells .METHODS: miR-155-specific siRNA and/or Ara-C were used to treat the cells .Quantitative real-time polymerase chain reaction was used to detect the expres-sion of miR-155.The growth of the cells was analyzed by CKK-8 assay.The cell apoptosis was determined by flow cytome-try.RESULTS:The miR-155 expression level of the cells transfected with miR-155 siRNA was significantly lower than that in the 2 control groups .Ara-C or miR-155 siRNA alone inhibited the growth of Raji cells in a dose-depend manner . miR-155 siRNA combined with Ara-C produced more inhibition of cell proliferation (P<0.05).After treatment for 48 h, the apoptotic rate of Raji cells in miR-155 siRNA+Ara-C group [(38.4 ±1.4)%] was higher than that in Ara-C group [(16.5 ±0.3)%] and miR-155 siRNA group [(14.6 ±0.3)%], with statistically significant difference (P<0.05). The expression of caspase-3 in Ara-C+miR-155 siRNA group was increased significantly as compared with Ara-C group and miR-155 siRNA group.CONCLUSION:miR-155-specific siRNA enhances the chemosensitivity of Raji cells to Ara-C by inducing apoptosis through the caspase-3 pathway .

To study the action, characteristics and expression of high methylation of p15(INK4B) and p16(INK4A) genes in multiple myeloma (MM), the sensitive methylation specific PCR method was employed to detect the hypermethylation of p15(INK4B) and p16(INK4A) in 24 patients with MM. Results showed that the methylation incidence of p15(INK4B) and p16(INK4A) genes were 70.8% (17/24) and 58.3% (14/24) in the MM patients, with the products of 148 bp and 150 bp fragments, respectively. The methylation of p15(INK4B) and p16(INK4A) genes were simultaneously happened in MM patients of plasmocytoma type with two cases at II phase and two cases at III phase. The simultaneous non-methylation of p15(INK4B) and p16(INK4A) genes were founded in five cases of MM patients, all of the tumor cells were of small plasmocyte type with mature differention. Conclusion suggested that there were high incidence of methylation of p15(INK4B) and p16(INK4A) genes in patients with MM. Hypermethylation can be detected in the early stage of disease, which was associated with its progress. It indicated a bad prognosis when methylation happended simultaneously in the two genes. Methylation of p15(INK4B) and p16(INK4A) genes may be related to the pathogeny of MM.