Objective:To clarify the effect of voltage-gated proton channel 1 (Hv1) on the migration and invasion of breast cancer cells. Methods:The protein expression of Hv1 was detected in human breast cancer cell lines with different metastatic abilities. SiRNA technique was used to down-regulate the expression of Hvl in breast cancer MDA-MB-231 cells. Scratch and matrigel invasion methods were used to observe the effect of Hvl on the migration and invasion of breast cancer cells, and the relevant molecular mechanism was explored. Results:Hv1 was highly expressed in the highly metastatic breast cancer cell line MDA-MB-231. Hvl was more highly expressed in MDA-MB-231 cells with higher metastatic ability. The SiRNA sequence target at Hvl inhibited Hvl expression. Scratch and matrigel invasion experiments showed that the migration and invasion of MDA-MB-231 cells were significantly attenuated when Hv1 was knocked down by siRNA targeting Hv1. Zymography experiment on matrix metalloproteinase indicated that the enzyme activities of MMP-2 markedly decreased. Conclusion:Hv1 promoted the migration and invasion ability of breast cancer cells.

This study was aimed to investigate the effects of resveratrol on plasma glucose and the oxidative stress ability in gestational diabetic rats. A total of 100 SD rats with gestation for 5 days were intraperitoneally injected with streptozotocin (STZ, 35 mg?kg-1) to prepare gestational diabetic rat model. Rats were randomly divided into 6 groups, which were the normal gestation group, gestational diabetic model group, resveratrol (30, 60, 120 and 240 mg?kg-1) treatment groups, with 20 rats in each group. A total of 20 rats with gestation for 5 days were selected as the normal gestation control group and another 20 rats with no gestation were selected as the normal control group. The levels of plasma glucose and insulin were determined on 0, 7, 14 days after experiment. Two weeks later, the contents of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) in serum were determined. The results showed that compared with the normal control group, there were no significant difference on the levels of plasma glucose and insulin, the content of MDA, and the activities of SOD, GSH-Px and CAT of the normal gestation control group (P > 0.05). Compared with the gestational diabetic model group, the content of MDA in serum of the resveratrol (30, 60, 120 and 240 mg?kg-1) treatment groups were significantly decreased (P < 0.05,P < 0.01); the level of plasma glucose were significantly decreased and the level of insulin was significantly increased of the resveratrol (120 and 240 mg?kg-1) treatment groups (P < 0.05,P < 0.01); the activities of SOD, GSH-Px and CAT were also significantly increased (P < 0.05, P < 0.01). It was concluded that resveratrol had dose-dependent effect on reducing plasma glucose and improving antioxidant ability in gestational diabetic rats, which perhaps related to its effects on raising the level of insulin and improving the activity of antioxidant enzymes.

Objective:To assess the expression levels of CD95 and OX40 ligand(OX40L)messenger RNA(mRNA)in hepatocellular carcinoma(HCC),and their clinical values thereof.Methods:The lymphocytes of research objects were mixed with corresponding fluorescence labeled monoclonal antibody(McAb);CD95 expression by CD3 positive T ceils was quantitatively measured by dual color flow cytometry.The expression of OX40L mRNA was detected in peripheral blood mononuclear cells by fluorescence quantitative reverse transcription polymerase chain reaction(FQ-RT-PCR).Results:The CD95 expression by CD3 positive T cells was significantly higher in patients with HCC(34±20)% compared with that of normal controls (20±7)%(t=2.960,P ＜ 0.01).The expression level of OX40L mRNA was significantly decreased in patients with HCC compared with that of normal controls(t=2.302,P ＜ 0.05).Conclusion:These results suggest that abnormal expressions of CD95 and OX40L play crucial roles in peripheral blood of HCC patients at the stage of human hepatocareinogenesis.

Aim To investigate the role of Wnt/β-cate-nin signaling pathway on the baicalin-induced osteo-genic differentiation in rat bone marrow derived mesen-chymal stem cells ( rBMSC ) . Methods rBMSC was isolated and cultured by adherence screening method. Alkaline phosphatase ( ALP) amount, CFU-FALP and mineralized nodules were compared between each ba-icalin group and vehicle control group at different time points. Real time q-PCR was employed to evaluate the mRNA level of Wnt signaling-related marker ( Wnt10a, GSK-3β,β-catenin and LEF1) after baica-lin treatment. Protein expression of β-catenin and Runx2 was measured by Western blot. Results Ba-icalin significantly increased ALP activities from day 3 to day 7 . The formation of CFU-FALP and mineralized nodules remarkably increased after rBMSC was treated with1, 10, 50 μmol · L-1 baicalin. mRNA levels of Wnt10a, β-catenin, GSK-3β, LEF1and osteocalcin were enhanced significantly in baicalin-treated group compared to control group. Protein expression of β-catenin and Runx2 was also elevated. Conclusion Baicalin ( 0. 1 to 50 μmol · L-1 ) promotes the osteo-genic differentiation and maturation of rBMSC, in which Wnt/β-catenin signaling pathway might be in-volved.

Objective:To silence ERCC2 (excision repair cross-complementing rodent repair deficiency,complementatin group 2,ERCC2) expression in esophageal cancer KYSE150 cells by small interfering RNA (siRNA) and observe the altered sensitivity of KYSE150 cells to paclitaxel (PTX) and elucidate the mechanism underlying the reversion of the PTX resistance of KYSE150 cells. Methods:ERCC2-targeted siRNA was synthesized in vitro and transiently transfected into ERCC2 overexpressing KYSE150 cells via Lipofectamine mediation. The mRNA and protein expression levels of ERCC2 were determined by using RT-PCR and FCM method, respectively. The sensitivity of KYSE150 cells to PTX was measured by MTT assay before and after siRNA transfection. Results:RT-PCR results suggested that the specific bands of ERCC2 mRNA were not detected in si-ERCC2 group at 24, 48 and 72 h post transfection. FCM results indicated that the expression levels of ERCC2 protein gradually decreased by 31.2%, 51.6% and 60.0% at 24, 48 and 72 h respectively(P<0.01). The IC_(50) value of PTX for ERCC2-silencing KYSE150 cells was (6.32±0.87) μg/mL, lower than that for control cells (P<0.01). Conclusion:siRNA successfully silenced the expressions of target gene ERCC2 at both the transcription and translation levels. Silencing ERCC2 expression partly reversed the resistance of KYSE150 cells to PTX.

Objective To evaluate the safety and efficacy of bivalirudin during perioperation of percutaneous coronary intervention (PCI) in patients older than 80 years old with acute coronary syndrome. Methods A total of 64 patients were randomly divided into two groups, respectively received heparin(n=32), or bivalirudin (n=32). We compared the activated coagulation time (ACT), procedural success rate, bleeding rate between two groups. Results The two groups ACT, PCI success rates are not statistically different, No signiifcant difference in the incidence of mild hemorrhage which is 4 (12.5%) for heparin and 1(3.1%) for bivalirudin and severe bleeding which is 2 (6.2%) for heparin and 0 for bivalirudin. However there are signiifcant differences in the overall incidence of bleeding between the two groups with lower incidence of beeding in the bivalirudin group than the heparin group (P<0.05). Conclusions Bivalirudin has comparable anticoangulation effect as heparin during perioperation of percutaneous coronary intervention (PCI) among patients older than 80 years of age with acute coronary syndrome with lower bleeding incidence than heparin dose.

To explore novel antifatigue agents targeting with AMPA receptor, 10 compounds were synthesized and their structures were confirmed by 1H NMR, ESI-MS and elemental analysis. 1-BCP was treated as the leading compound. The antifatigue activities were evaluated by weight-loaded forced swimming test, and the AMPA receptor binding affinities were tested with radioligand receptor binding assays. The results unveiled that 5b appeared to possess potent antifatigue activities and high affinity with AMPA receptor, which deserved further studies.

Objective:To analyze the clinico-pathological characteristics, pathological diagnosis, and treatment of rhabdoid tu-mor. Methods:The medical records of four rhabdoid tumor patients that were admitted to the Tianjin Medical University Cancer Insti-tute and Hospital since 2000 were analyzed based on existing literature. Results:In one of the four cases, the tumor originated from the kidney, whereas in the other three, the tumor occurred from extra-renal soft tissues. Histologic analysis revealed that the tumor cells were loosely arranged with diffuse growth, vesicular nuclei, dyed cytoplasm, visible eosinophilic inclusions, and more nuclear fission. The results of immunohistochemical staining showed that the vimentin and epithelial membrane antigen were positive, whereas CK, CD99, CD34, and S-100 were positive at different degrees. MyoD1, Desmin, and INI-1 were negative. Conclusion:Rhabdoid tumor is rare and highly aggressive. It occurs mainly in the kidney and can also be found in other systems. The unique pathological form and im-munohistochemical staining observed on the tumor can be used as reference for diagnosis.

Background and purpose: ERCC2 gene silence by siRNA interference was observed in esophageal cancer cell line and it could change cell sensitivity to taxol. This study was to investigate the biological mechanism of paclitaxel-resistance in esophageal cancer cells. Methods: ERCC2 targeting siRNA (si-ERCC2) has been synthesized. The constructor were transfected into ERCC2 cell lines high-KYSE150(bigh expression of ERCC2) through lipofectamine. RT-PCR and flow cytometry (FCM) were used to detect the ERCC2 mRNA and protein expression levels. MTr assay was used to estimate the paclitaxel sensitivity of the cells. Results: In si-ERCC2 group, ERCC2 could not be detected and ERCC2 protein expression were reduced by 31.2%, 51.6% and 60.0%, respectively, 24,48,72 b after transfection. Paclitaxel IC_(50) value for si-ERCC2 group was 6.32±0.87 μg/mL, lower than the control group(49%). Conclusion: siRNA could successfully silence the target gene ERCC2 at the level of transcription and translation of the gene, the reduction of ERCC2 expression may reverse taxol resistance of the cells.

Objective To establish a method for isolating and culturing fibroblasts from cirrhotic liver in vitro,observe the biological characteristics of fibroblasts.Methods Human fibroblasts from cirrhotic liver were isolated and cultured by adhering to the culture plastic.The morphologic and growth characteristics of the acquired cells were observed by microscopy imaging and cell counting.The expression of FSP-1 and Vimentin of fibroblasts was detected through immunofluorescence and western blotting.Results The tissue blocks were well-adherent to the culture plastic within 2 hours.Hepatocyte-like cells were observed surrounding the blocks within about 1 week,and spindle-shaped fibroblasts were observed within about 2 weeks.After a series of passage,the attached cells became proliferated quickly.The growth curves of the passage 3,4 and 5 were quite similar.The result of immunofluorescence showed the expression of FSP1 and Vimentin was positive.The positive rate was respectively (90.6 ± 1.0) ％ and (91.2-± 4.1) ％.Conclusions Human fibroblasts from cirrhotic liver can be cultured successfully through adherent property.The method for isolation and culture of fibroblasts from cirrhotic liver is convenient,efficient,stable and cultured cells remain naive biological characteristics.