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.

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

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.

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

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

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 effect of microRNA-155(miR-155) on transforming growth factor β2 (TGF-β2)-induced epithelial-mesenchymal transition of human retinal pigment epithelial cells and its mechanism.Methods:The retinal pigment epithelial cell ARPE-19 cell line was used as the research object.The cells cultured with DMEM medium were served as the control group and the cells cultured with DMEM medium containing 10 ng/ml TGF-β2 were served as the TGF-β2 group.The ARPE-19 cells transfected with miR-155 inhibitor were set as the miR-155 inhibitor group and the ARPE-19 cells transfected with miR-155 negative control were set as the miR-155 negative control group, and the cells in the two groups were cultured in DMEM medium containing 10 ng/ml TGF-β2.After 48 hours cell culture, reverse transcription-PCR was used to detect the expression of miR-155 in each group, and scratch migration test and Transwell chamber test were used to detect cell migration and invasion ability, and Western blot was used to detect the expressions of phosphate and tension homology deleted on chromosome ten gene (PTEN), phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), p-Akt and epithelial mesenchymal markers E-cadherin (E-cad), zonula occludens protein 1 (ZO-1), F-actin, α-smooth muscle actin (α-SMA), fibronectin 1 (FN-l) vimentin, proteins.The target gene prediction library predicted miR-155 target gene and fluorescein enzyme reporter vectors were used to identify target genes.Results:After 48 hours of culture, the cells in the control group were in good condition with tight adherence and regular shape.The cells in the TGF-β2 group showed more obvious spindle shape with loose arrangement, and most of the cells were fibrous.The relative expression level of miR-155 in the cells of TGF-β2 group was 0.92±0.14, which was significantly higher than 0.35±0.06 of the control group ( t=7.242, P=0.003). The relative expression level of miR-155 in the cells of miR-155 inhibitor group was 0.21±0.03, which was significantly lower than 0.98±0.09 of the miR-155 negative control group ( t=12.421, P<0.01). The migration rate was higher and the number of cells passing through basement membrane was more in the TGF-β2 group than those of the control group, and the migration rate was higher and the number of cells passing through basement membrane of miR-155 was more in the miR-155 negative control group than those of the miR-155 inhibitor group, and the differences were statistically significant (all at P<0.01). Compared with the control group, the relative expression levels of PTEN, E-cad, ZO-1, F-actin protein were decreased, while the relative expression levels of PI3K and the p-Akt/Akt ratio were increased, and the relative expression levels of α-SMA, FN-1, vimentin proteins were increased in the TGF-β2 group, and the differences were statistically significant (all at P<0.01). Compared with the miR-155 negative control group, the relative expression levels of E-cad, ZO-1, F-actin and PTEN proteins were increased, while the relative expression levels of α-SMA, FN-l, vimentin, PI3K and the p-Akt/Akt ratio were decreased in the miR-155 inhibitor group, and the differences were statistically significant (all at P<0.01). Target gene prediction library prediction and luciferase reporter vector identification confirmed that PTEN was a downstream target gene of miR-155. Conclusions:miR-155 can promote the TGF-β2-induced epithelial-mesenchymal transition progress in human retinal pigment epithelial cells, and its mechanism may be related to inhibiting the expression of the target gene PTEN and stimulating the activation of the PI3K/Akt signaling pathway.

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 investigate the feasibility of injection of sodium hyaluronate gel to upper lacrimal puncta in locating the nasal broken end of inferior canalicular laceration.Methods Together 52 patients(52 eyes) with inferior canalicular laceration who collected from March 2013 to March 2016 in the Third Affiliated Hospital of Nanchang University underwent canaliculax laceration anastomosis combined with silicone tube.Injection of sodium hyaluronate gel to the upper lacrimal puncta was introduced in group A (n =32/32 eyes) for locating the nasal broken end of lower canaliculax laceration,while microscope for searching the nasal broken end served as group B (n =20/20 eyes).In both groups,the silicone tubes were implanted in the lacrimal passage for more than 3 months after locating the broken end successfully.All patients were followed up from 6 months to 12 months,with mean follow-up of (8.2 ± 1.6) months,and then clinical data of the distance of lower canalicular laceration between lacrimal canal,time consuming for locating the nasal broken end,effective rate and postoperative comphcations were compared between two groups.Results The procedures in all patients were successfully.In group A,23 patients were cured and 5 improved,but the operation was failed in 4 patients,of which 3 patients occurred upper or lower lacrimal punctas teax;while in group B,13 patients were cured and 4 improved,but the operation was failed in 3 patients,of which 2 patients occurred upper or lower lacrimal punctas tear.There was no significant difference in the the distance of between lower canalicular laceration and lacrimal canal in group A [6.5-8.3 (7.3 ± 0.6) mm] and group B [6.6-8.2 (7.2 ± 0.5) mm] (P =0.40).The time consuming for locating the nasal broken end in group A [1.5-5.5 (3.3 ± 1.3) min] was shorter than that in group B [5.0-26.0 (17.0 ± 6.0) min],with significant difference (P ＜ 0.001).Conclusion It is an easy and accurate method of injection of sodium hyaluronate gel to upper lacrimal puncta for locating the nasal broken end of lower canalicular laceration with less time.