ObjectiveTo investigate the effects of recombinant human erythropoietin (rhEPO) on the apoptosis of human mesenchymal stem cells induced by H2O2 and related cell signaling pathways.Methods After being cultured in vitro, human MSCs were treated with rhEPO.Phosphorylation of ERK1/2,p38 MAPK and PI3K/Akt was detected by using Western blotting.One h after pretreatment with1U/ml rhEPO,cells were cultured in the presence of1mmol/L H2O2 for1h.Cell morphology was observed under the inverted microscopy.Cell apoptosis was determined by using flow cytometry,migration assay was performed in Transwell chambers,and adhesion assay was performed by plastic dishes.ResultsRhEPO could increase phosphorylation of PI3K/Akt pathway in human MSCs,but reduce phosphorylation of p38MAPK.RhEPO had no obvious effect on ERK1/2 pathway and total proteins of Akt and p38MAPK.RhEPO could decrease apoptosis of human MSCs induced by H2O2 (P＜0.01) and the inhibitory effect was abrogated by Ly294002 but not anisomysin.Conclusion RhEPO could protect MSCs from apoptosis. Activation of PI3K/Akt pathway was involved in the effect of rhEPO on apoptosis.

Objective To explore the changes of CD4~+ cell and relevant cytokines from chronic hepatitis B patients and the significance was discussed. Methods The expression of CD4,CD8 and CD28 molecules on PBMC was detected with Flow Cytometry.The levels of IL-2,IFN-?,IL-4,IL-10 and IL-12 in serum were measured with ELISA,and the liver function test including serum ALT,SB,ALP and albumin was conducted. Results Three conditions of CD4~+ ratio from PBMC in 86 patients were observed,that showed lower,similar or higher comparing with normal controls.According to CD4~+T cell ratio,the patients were divided into 3 groups.In three groups,the changes of CD28 expression paralleled to that of CD4~+T cell ratio.Except IL-4,the level of cytokines related to viral clearance such as IL-2,IFN-?,IL-10 and IL-12 in patients was elevated compared with controls(P

Objective To investigate factors and neonatal outcomes associated with histologic chorioamnionitis(HCA) after preterm premature rupture of membranes (PPROM).Methods From January 2008 to January 2013,103 women with PPROM at 28-33+6 weeks of gestation undergoing deliveries were studied retrospectively.According to placental histopathologic findings,those patients were categorized into two groups,including 68 cases in histologic chorioamnionitis (HCA group) and 35 cases in non-chorioamnionitis (control group).Age,parity,gestational age of PPROM and delivery,latency period,oligohydramnios,white blood cell (WBC) count and serum C reactive protein (CRP) level at admission and before delivery,and the neonatal outcomes were compared between two groups.The risk factors were analyzed by multivariable Logistic regression analysis.Results The incidence of HCA was 66.0％ (68/103) in all cases with PPROM.The occurring ruptured membrane gestation in HCA group was (28.2 ± 1.2) weeks,which were significantly earlier than (32.3 ± 1.4) weeks in control group (P ＜ 0.05).The level of CRP of (8.3 ± 4.7) mg/L before dehvery in HCA group was significantly higher than (5.4 ± 3.2) mg/L in control group (P ＜ 0.05).The rates of oligohydramnios and cesarean sections were significantly higher than those in control group (P ＜0.01 or ＜0.05).Using multivariable Logistic regression analysis,oligohydramnios,gestational age of PPROM ＜ 32 weeks,serum CRP level ＞ 8 mg/L before delivery and latency period 48-168 h were significantly associated with HCA (P ＜ 0.01 or ＜ 0.05).The gestational age of delivery and birth weight of HCA group were significantly lower than those of control group (P ＜ 0.05).The incidence of Apgar ＜ 7 scores,abnormal brain ultrasonography findings,neonatal pneumonia,bronchopulmonary dysplasia,early-onset neonatal sepsis and mortality in HCA group were significantly higher than those in control group (P ＜ 0.05).Conclusions HCA has significantly correlated with lower gestational age of PPROM,higher serum CRP level before delivery,prolonged latency period and oligohydramnios in PPROM.HCA could increase the neonatal morbidity and mortality.

Aim To observe effects of breviscapine on lymphocyte proliferation and K562 cell death caused by doxorubicin.Methods The cell proliferation was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay,the p53/bcl-2 expression were assessed by western blotting,cell apoptosis was quantitated by Histone/DNA ELISA and flow cytometry,cytochrome C release was determined by ELISA,caspase-8 and caspase-3 activation were examined by colorimetric assay.Results Breviscapine could obviously enhance mouse lymphocyte proliferation and its resistance to doxorubicin,promote growth inhibition,cytochrome C release,caspase-8 and caspase-3 activation in K562 cells which were caused by doxorubicin,upregulate radio of p53/bcl-2 expression,and increase cell apoptosis.Conclusion Breviscapine was able to enhance antitumor effect of doxorubicin,the major mechanism by which breviscapine could promote sensitivity of tumor cell to chemotherapeutic agents might be related to activated apoptosis signal way.

ObjectiveTo investigate the expression of stem cell marker adenosine triphosphate (ATP)-binding cassette transporter 2 (ABCG2) in the tissue and side population (SP) of a cell line A431 of human cutaneous squamous cell carcinoma(SCC).MethodsSP cells were separated by flow cytometry from cultured A431 cells.Methyl thiazolyl tetrazolium(MTT) assay was used to evaluate the proliferative ability of,reverse transcription-PCR to determine the expression of ABCG2 in,SP and non-SP cells.Immunohistochemistry (MaxVision method) was carried out to detect the expression of ABCG2 protein in tissue specimens from 10 patients with SCC.ResultsSP cells existed in cultured A431 cells,and accounted for about 1.1％ of A431 cells.The SP cells had a stronger growth and colony-forming ability than non-SP cells did.The number of cell clones formed by SP cells and non-SP cells was 114.8 ± 4.95 and 44.5 ± 3.67,respectively,per well in a 24-well plate( t =27.92,P ＜ 0.01 ).The expression level of ABCG2 mRNA was significantly higher in SP cells than in non-SP cells(t =5.22,P＜ 0.01).There existed a small number of ABCG2 positive cells in SCC tissue,and ABCG2 was mainly expressed in the cytoplasm and membrane of tumor cells.ConclusionsSP cells exist in A431 cells,which have characteristics of stem cells and highly express ABCG2.ABCG2 may be a potential stem cell surface marker of SCC.

BACKGROUND: Results from clinical trials suggested that clopidogrel and ticlopidine had side effects of granulopenia, and aspirin could inhibit endothelial progenitor cell proliferation. There is no report of effects of these drugs on human bone marrow mesenchymal stem cells (hBMSCs) in stem cell transplantation. OBJECTIVE: To investigate the effects of antiplatelet drugs including clopidogrel, ticlopidine and aspirin on hBMSC proliferation and secretion. DESIGN, TIME AND SETTING: The cytology in vitro observation was performed at the Laboratory of Toxicology, Shanghai Municipal Center for Disease Control and Prevention from March to December 2006.MATERIALS: The second passage of hBMSCs was kindly donated from Shanghai Tissue Engineering Research & Development Center, Shanghai Ninth People's Hospital. Clopidogrel (Lot number J20040006) and ticlopidine (Lot number H19980186) were obtained from Hangzhou Sanofi-Synthelabo Minsheng Pharmaceutical CO., Ltd. Aspirin (Lot number 20050059) was obtained from Bayer Vital GmbH. METHODS: The standard culture medium consisted of DMEM-LG, 10% heat-inactivated FBS, 100 U/mL penicillin and 100 μg/mL streptomycin. After being cultured in vitro expanded out to passage 6, hBMSCs were treated with antiplatelet drugs of different concentrations and compared with control group. MAIN OUTCOME MEASURES: Cell proliferation was assessed by 3- (4, 5-dimethylthiazol -2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay, level of vascular endothelial growth factor (VEGF) of culture medium was detected by enzyme-linked immunoadsordent assay (ELISA), and surface antigens of hBMSCs were analyzed by the flow cytometry. RESULTS: A570 values of hBMSCs treated by clopidogrel or ticlopidine of 0.02,0.1,0.4,2,10,40 μmol/L were higher than control group (P < 0.01), while A570 values of aspirin group of 60, 600, 2 000 μmol/L were lower than control group(P < 0.05). Antiplatelet drugs had no obvious effect on cell surface antigens(CD34, CD105, CD106)expressed by hBMSCs. Treated by high dose clopidogrel or ticlopidine (40 μmol/L), VEGF level from hMSCs was lower than that of control group(P < 0.01), but VEGF level of low dose (0.02 μmol/L) ticlopidine group was higher than control group(P < 0.01), and there was no significantly difference of VEGF level among low dose clopidogrel group (0.02 μmol/L), aspirin group (5, 2 000 μmol/L), and control group(P > 0.05). CONCLUSION: Clopidogrel and ticlopidine improve proliferation of hBMSCs, but aspirin inhibits proliferation of hBMSCs. High dose of clopidogrel and ticlopidine suppress VEGF secretion of hBMSCs, while low dose of ticlopidine promote it. Antiplatelet drugs have no obvious effect on hBMSCs differentiation.

Objective To analyse the effects of anastomat to the resection surgery in 1800 esophageal and gastric cardiac carcinoma patients. Methods The Esophagus-gaster and Esophagus-intestine were stapled by anastomat in the cervical region in 182 cases、 intrathoracically in 1296 cases and intraperitoneal in 322cases. The occurrence of complications caused by anastomat, including anastomotic fistula,anastomotic stricture,anastomotic bleeding and mechanical failure,were observed. Results Anastomotic fistula occurred in 15 cases ( 15/1800,0.83% ,ten cases took Shanghai-made GF-I anastomat ,five cases took YH-W single disposable single anastomat ), among which 6 cases had the cervical anastomosis; Anastomotic stricture occurred in 41 cases ( 41 /1800,3.11%, fifteen cases took Shanghai-made GF-I anastomat, twenty-six took YH-W single disposable single anastomat) ,but all of them recovered after dilatation; Anastomotic bleeding occurred in 21 cases (21/18001.16%, thirteen cases took Shanghai-made GF-I anastomat, eight took YH-W single disposable single anastomat) ;Anastomat mechanical failure in operation occurred in 14 cases( 14/1800,0. 78% ,ten cases took Shanghai-made GF-I anastomat, four took YH-W single disposable single anastomat). Conclusion Anastomat is an effective method in reducing the postoperational complications of esophageal and gastric cardiac carcinoma resection. Disposable single anastomat has higher clinical value.

Objective To compare the concentration of nuclear factor-kappa B (NF-κB), MCP-1 and RANTES in peritoneal fluid in women with endometriosis and normal peritonal fluid ,To explore the relationship between these cytokines with pathogenesis of endometriosis. Methods There were 2 groups:the endometriosis group and the control group.Double-antibody sandwich ABC-ELISA technique was adopted for determinating concentration of NF-кB、MCP-1 and RANTES in peritoneal fluid. Results Peritoneal fluid level of NF-κB MCP-1 and RANTES in endometriosis patients was higher than the control group (P < 0.05). NF-κB and MCP-1, NF-κB and RANTES expressed positive correlation by linear correlation analysis in both EMs group and control group , correlation coefficients were 0.385, 0.569(EMs group), 0.474, 0.388(P < 0.05). Conclusions NF-кB、MCP-1 and RANTES are mutual affection and promote endometriosis in the pathogenesis.

Aim To observe the protective effects of breviscapine against lung fibrosis and investigate its possible mechanism.Methods Effects of breviscapine on cell proliferation,activation and extracellular matrix secretion were examined in mouse embryonic lung fibroblast L929 cells in vitro.The mouse model of bleomycin-induced lung fibrosis was used to assess the protective effect of breviscapine against lung fibrosis.Results In vitro,breviscapine had no cytotoxicity directly on L929 cells,however,it could suppress cell proliferation,activation and secretion of laminin(LN) and collagen Ⅰ(ColⅠ) induced by transforming growth factor beta1(TGF-?1) in L929 cells.In vivo,breviscapine could prevent increase in serum TGF-?1 and decrease in superoxide dismutase(SOD),peroxidase(POD) and catalase(CAT) in mice with lung fibrosis caused by bleomycin.In addition,breviscapine was able to reduce pulmonary hydroxyproline,collagen,malondialdehyde(MDA)and TGF-?1 in lung fibrosis mice.Conclusion Breviscapine has protective effect against lung fibrosis and the possible mechanism is to enhance antioxidative defense activities and prevent TGF-? signal.

Aim To study antiangiogenesis and antitumor of thalidomide.Methods In HUVECs, cell viability was determined by MTT assay;death type was observed by electron microscope; ratio of apoptosis was quantitated by flow cytometry.Angiogenesis was tested in chicken embryo chorioallantoic membrane.Effect of thalidomide on S_(180) was examined in homograft mice and microvascular counts were detected through immunochemical staining method.Results Thalidomide might inhibite the growth of HUVECs with a IC_(50) value of(22.91?1.74) ?mol?L~(-1),cells treated by thalidomide for 48 h displayed morphological characteristics of different stages associated with apoptosis,which were irregular nucleus, condensed chromatin, ballooning endoplasmic reticulum, apoptotic bodies,under electron microscope.Thalidomide might be able to cause apoptosis or necrosis of HUVECs in flow cytometry and raised positive of antiangiogenesis with increasing of dosage in chicken embryo chorioallantoic membrane. Thalidomide as a single agent might not significantly prevent tumor growth but decrease microvascular counts in tumors, however, in combination with cyclophosphamide, thalidomide could decrease dosage of cyclophosphamide and enhance antitumor of cyclophosphamide.Conclusion Thalidomide might hold back angiogenesis,as a single agent, could not significantly prevent S_(180) tumor from growing,but acted synergistically with cyclophosphamide.