Objective To optimize multiplicity of infection ( MOI) and antibiotics ( blasticidin) concentration selecting BSD gene in construction of monoclonal stable cell line by lentivirus vector-mediated RNA interence silenced gene SGMS2.Methods The INS-1 cells were transfected by fluorescence labeled negative control SGMS2-siRNA lentivirus at MOI of 0, 10, 30, 60 and 120 TU number/cell.The cells were photographed under fluorescent microscopy after 72 h cultivation, then fluorescence ratio and apoptosis rate were calculated to determine optimal MOI.The INS-1 cells were treated by blasticidin with different concentrations of 0, 1, 2, and 3 μg/mL, and the apoptosis rate was observed to acquire optimal concentration of antibiotics.The INS-1 cells were transfected by negative control SGMS2-siRNA lentivirus and SGMS2-siRNA lentivirus (virus titer:1 ×108TU/mL) at optimal MOI and positive-transfected cells were selected by blasticidin at optimal concentration, then mixed cell lines were acquired.The monoclonal cell line was constructed at fluorescence ratio of 90%.Results The optimal MOI was 60 with 100% fluorescence ratio, less than 0.5% apoptosis rate and keep original cellular morphology.The optimal concentration of blasticidin was 2 μg/mL with cell adherence disappear and all cells apoptosis.The Ct value of INS-1-SEMS2 cells detected at the second time was 28.21, which was greater than 27.58 at the first time.The interfering efficiency of siRNA was 77.78% which indicated a successful expression of siRNA and construction of monoclonal stable cell line ( INS-1-SEMS2 ).Conclusion The monoclonal stable cell line was successfully constructed by lentivirus vector-mediated RNA interence silenced gene SGMS2.

Objective To investigate the relationship between the p27~(kip1) expression and the change of radiosensitivity of esophageal cancer cells.Methods Radio-resistant cells (EC9706R) were gradually isolated by means of repeated gamma-ray irradiation (6 000 cGy in total) upon human squamous esophageal carcinoma cells (EC9706).The radiosensitivity of these two types of cells was measured by standard colony formation assays,their cell cycle distribution analyzed by flow cytometry respectively,and the p27~(kip1) expression detected by immunocytochemistry.Results As compared with their parent cells,the isolated human squamous esophageal carcinoma cells(EC9706) showed clearly greater radio-resistance(for EC9706R,SF_2=65.71%,D_0=2.20 Gy,D_q=1.61 Gy,N=2.07,and for EC9706,SF_2=46.72％,D_0=1.61 Gy,D_q=0.99 Gy,N=1.85;SF_2,D_0,Dq and N were all higher).As to the cell cycle distribution,the population of G_1 phase cells in EC9706R cells was significantly decreased,but the proportion of S phase cells was significantly increased as compared with the parent cells.Immunocytochemistry revealed that the p27~(kip1) expression of EC9706R cells was clearly lower than that of EC9706 cells.Conclusion Cell phase change due to the decrease of p27~(kip1) expression may be one of the mechanisms of the generation of radio-resistance in esophageal cancer cells.

Objective To investigate the changes and its significance of expression of platelet CD62p,CD42b and the levels of plasma TXB2,6-keto-PGFla and vWF in patients with intracerebral hemorrhage.Methods 46 patients with intracerebral hemorrhage were included as the intracerebral hemorrhage group,while 20 patients with hypertension were served as the control group.Platelet CD62p and CD42b were measured by flow cytometry,and plasma TXB2,6-keto-PGF1a and vWF were measured by ELISA at less than 3 day,1 and 2 weeks after onset.The analysis of dependablit were made between the expression of platelet CD62p,CD42b and the level of plasma vWF in patients with intracerebral hemorrhage.Results At less than 3 day,1 and 2 weeks after onset,the expression of platelet CD62p,levels of plasma TXB2,vWF and TXB2/6-keto-PGFla(T/K)in patients with intracerebral hemorrhage were significantly higher than those in the control group(all P

Acute pancreatitis,a multiple system disease was associated with distant organs injury such as the liver,lung and kidney,etc.Acute renal failure is one of the major death causes in severe acute pancreatitis.Renal injury in acute pancreatitis and its pathogenesis was reviewed to further understand the mechanisms of systemic inflammatory response and multiple organ failure

Objective To explore the expression of estrogen receptor(ER) and progesterone receptor(PR) in gestational trophoblastic tumor(GTT) and their significance.Methods The expression of ER and PR in 34 cases of GTT was detected by immunohistochemical method;20 cases of normal villi and 30 cases of hydatiform mole served as controls.Results The positive expression rate of ER in normal villi,hydatiform mole and GTT was 85.00%,83.33% and 44.12%,respectively,and had positive correlation with the malignance degree of GTT.The positive expression rate of PR in normal villi,hydatiform mole and GTT was little.The expression of ER was closely related to these clinicopathological features of GTT(P

1cm)(P=0.0000)and the type of adjuvant chemotherapy implemented (non-standard chemotherapy versus standard chemotherapy)( P=0.0028). Among them tumor size was exclusively an independent factor.Conclusions:Histology、FIGO stage、residual tumor after surgery and the type of adjuvant chemotherapy were independent factors associated with recurrence in MOGCT .For those patients with high risks of recurrence, full dose and cycles of standard BEP/PVB regimen should be used.

Clinical data of 61 cases of imported malaria were analyzed retrospectively.Patients with malaria were divided into three groups,asymptomatic tertian (vivax) malaria,symptomatic tertian malaria and pernicious (falciparum) malaria.Only mild hepatic damage occurred in some patients with asymptomatic tertian malaria,compared with other groups Symptomatic tertian malaria and pernicious malaria were misdiagnosed in 6 of 20 and three of six,respectively before hospitalization,and 16 of 20 and four of six patients complicated with thrombocytopenia,respectively,and both of them had increased serum level of C-reactive protein (CRP).Platelet count negatively correlated with their serum level of CRP significantly in patients with symptomatic tertian malaria (r =-0.555,P ＜ 0.05).Routine anti-malaria therapy was used in imported malaria,blackwater fever occurred in two patients and acute renal failure occurred in one with falciparum malaria.It is suggested plasmodium exam ination in peripheral blood should be performed in all persons returned from countries prevalent with malaria,thrombocytopenia is an indicator of acute malaria,and more severe complications usually tend to occur in falciparum malaria.

Objective To investigate the expression and distribution of fibroblast growth factor-13(FGF-13) in rat vibrissa follicle at anagen and the cultured cells derived from bulge region.Methods FGF-13 expression was detected using immunohistochemistry and immunocytochemistry.Results The in vitro cultured cells from bulge region and the vibrissa follicle expressed FGF-13.In vivo,the cells expressing FGF-13 distributed in the outermost layer of the outer root-sheath(ORS) in the isthmus portion of the follicle.No cells expressing FGF-13 were present in the hair follicle bulb,including matrix cells and ORS.Conclusion In vibrissa follicle at anagen FGF-13 may be involved in the migration of stem cells located at the bulge region to the hair bulb.

Objective To observe the effect of triptolide combined with gemcitabine on proliferation and apoptosis of pancreatic cancer cells ,and to analyze the relevant mechanisms .Methods After treated with TPL ,GEM or TPL combined with GEM in vitro , PANC-1 cells proliferation was accessed by MTT assay and the interaction between the two drugs was calculated .Apoptotic mor-phological changes and apoptosis rate of the cells were investigated by Hoechst 33258 staining and flow cytometry ,respectively .The expression of signal transduction and transcription factor 3(STAT3) ,cysteine aspartate specific proteases-3(caspase-3) protein were detected by Western blot analysis .Results TPL ,GEM or TPL combined with GEM could significantly inhibit the prolifera-tion of PANC-1 cells ,and the combination of the two drugs had a synergistic effect .The cells of the TPL group ,GEM group ,as well as the combined group showed typical apoptotic morphological changes .Compared with the TPL group and GEM group ,the number of apoptotic cells of the combined group increased significantly .Compared with the control group ,the cells apoptosis rate of the TPL group ,GEM group and combined group was significantly increased (P< 0 .05) ,and the apoptosis rate of the the combined group was significantly higher than that of the monotherapy group(P<0 .05) .TPL combined with GEM synergistically inhibited p-STAT3 protein expression and activated caspase-3 protein expression .Conclusion TPL combined with GEM can synergistically in-hibit proliferation and induce apoptosis of pancreatic cancer PANC-1 cells ,its mechanism is related to the inhibition of STAT 3 sig-naling pathway ,promotion the expression of caspase-3 protein .

Based on the results of oral glucose tolerance test( OGTT )and the levels of 1-h plasma glucose ( 1 hPG),793 subjects were classified into three groups:583 with NGTN ( normal 1 hPG in OGTT),127 with NGT1 H( higher 1 hPG in OGTT) and 83 with IGT( impaired glucose tolerance).NGT1H group had large waist circumference,higher body mass index,fasting plasma glucose( FPG),triglyceride,and lower high density lipoprotein-cholesterol than those of NGTN group.NGT1 H group had higher homeostasis model assessment insulin index ( 1.2 ± 0.6),lower homeostasis model assessment β3 ( HOMA-β ) (4.5 ± 0.7 ) and insulinogenic index (2.1 ±0.7) than those of NGTN group(0.5 ±0.6,4.8 ±0.7,2.7 ±0.9,respectively,all P ＜0.05 ).HOMA-β of NGT1 H group was higher than that of IGT group(4.5 ±0.7 vs.4.4 ±0.6,P ＜0.05 ).The results indicate that 1 hPG in OGTT may identify a condition of glucose metabolic abnormalities characterized by insulin resistance and reduced β-cell function.