This study is to establish a cell-based model targeting to neuraminidase (NA) of the 2009 H1N1 influenza A virus. NA is an influenza virus structural protein with enzymatic activity of the cleavage of HA-sialic acid interaction to release new viral particles from cells. A model of HIV-1 (pNL4-3.Luc.R(-)E(-)) based pseudovirions packed with HA [hemagglutinin, A/VietNam/1203/2004 (H5N1)] and NA [A/California/04/2009 (H1N1)] was established to evaluate compounds activities on NA function. The viral release can be blocked by neuraminidase inhibitors, oseltamivir and oseltamivir carboxylate, with IC50 of (61 +/- 31) nmol L(-1) and (5.5 +/- 2.9) nmol L(-1) respectively. A point mutation of H275Y on NA leads oseltamivir-resistance. This corresponding mutation was introduced into the system which was also confirmed by oseltamivir and oseltamivir carboxylate.

BACKGROUND:Photoelectric navigation-aided percutaneous pedicle screw placement has been developed extensively,but its accuracy,safety and effectiveness have not yet been confirmed by evidence-based medicine.OBJECTIVE:To compare the curative efficacy of photoelectric navigation-aided percutaneous pedicle screw placement and traditional open posterior pedicle screw fixation for thoracolumbar fractures.METHODS:Sixty patients with thoracolumbar fractures were equivalently randomized to treatment and control groups and then underwent photoelectric navigation-aided percutaneous pedicle screw placement and traditional open posterior pedicle screw fixation,respectively.The perioperative indexes,imaging indexes,function recovery and incidence of complications were compared between two groups.RESULTS AND CONCLUSION:(1) The Visual Analogue Scale scores,intraoperative blood loss,radiant times,and hospitalization time in the treatment group were significantly less than those in the control group (P ＜ 0.05).(2) The operation time did not differ significantly between two groups (P ＞ 0.05).(3) The postoperative sagittal Cobb angle,and percentage of anterior height in the vertebral body in the two groups were significantly improved compared with those before surgery (P ＜ 0.05),but all above imaging indexes showed no significant differences between two groups (P ＞ 0.05).The endplate-screw angle in the treatment group was significantly less than that in the control group (P ＜ 0.05).(5) The excellent and good rate of placement in the treatment group was significantly higher than that in the control group (P ＜ 0.05).(6) These results suggest that compared with the traditional open posterior pedicle screw fixation,the photoelectric navigation-aided percutaneous pedicle screw placement exhibits high placement accuracy,less radiant times,less trauma,less blood loss and rapid functional recovery.

Objective To investigate the influence of hypoxia on leptin expression in serum,CSF and brain tissue of C57BL/6J mice.Methods C57BL/6J mice were divided into 3 groups:the normal control,24-hour hypoxia group and 48-hour hypoxia group.The latter two groups exposed to hypoxia in the man-made auto pressure and hypoxia control cabin(XQ-I).Radioimmunoassay was used to detect the leptin level.Results The leptin level in the serum,CSF and brain tissue of mice exposed to hypoxia were higher than that of normal control,but no difference was found between 24-hour hypoxia group and 48-hour hypoxia group.Conclusion Hypoxia probably induces the increasing of leptin expression in fat tissue,as well as in the central nervous system.

Objective Protein N-SRCR derived from salivary agglutinin (SAG) inhibits HIV-1 infection.An N-SRCR monoclonal antibody was prepared for the study of the interaction between N-SRCR and HIV-1 envelop glycoprotein (gp120).Methods The purified recombinant N-SRCR expressed by 293 cells was used to immunize four weeks old BALB/c mice.After the final boost,the mouse spleen cells were isolated and fused with mouse myeloma cell line SP2/0-Ag-14,and the resulting hybridomas were screened for the production of N-SRCR-specific antibodies by ELISA assay.The monoclonal antibody against N-SRCR was purified by HiTrap Protein G kit,the purity determined by SDS-PAGE and the antibody titers by ELISA.The antibody specificity.was charqacterized by western blotting.Results A strain of hybridoma cell clones stably secreting N-SRCR antibody,named 1D6,was obtained.The high purity of the IgG was demonstrated by SDS-PAGE,and the ELISA titers of 1D6 was more than 100?25.Conclusion A monoclonal antibody against N-SRCR was successfully prepared,which laid the ground for further studies on the biological function of N-SRCR and the interactions between SRCR domains and HIV-1 Env gp120.

Objective To observe the effects of ( - )-epigallocatechin-3-gallate (EGCG) on human prostate cancer xenografted tumor growth and connexin43 expression in nude mice,and explore the mechanism of the EGCG on prevention for prostate cancer.Methods The methyl thiazolyl tetrazolium and annexin-V/PI double-labeled flow cytometry methods were used to observe the growth inhibiting rate (IR)and apoptosis rate (AR) of human prostate cancer cell line PC-3 which was treated by EGCG at different concentration (10,20 and 40 mg/L,respectively).The scrape-loading fluorescence dye transfer method was applied to assess the gap junction intercellular communication (GJIC) through fluorescence microscope.PC-3 cells were subcutaneously transplanted to establish tumor-bearing nude mice model.A total of 32 mice were randomly divided into four groups,both control group and three treatment groups were treated with different doses of EGCG ( 10,20 and 40 mg/kg,respectively).After two weeks,the mice prostate tumor tissues were taken out.The tumor wet weight was measured and tumor growth inhibiting rate was calculated.The tumor microvascular density (MVD) and apoptosis index (AI) were detected by the immunohistochemical techniques and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling techniques,respectively.Semi-quantitative reverse transcription polymerase chain reaction was used to examine the expression level of the Cx43 mRNA.Results EGCG at concentration ( 10 and 20 mg/L) could significantly inhibit the proliferation[(22.33 ±4.62)％,(38.67 ±5.67)％ vs (3.47 ±0.31 )％,P ＜0.01],induce the apoptosis [(7.84 ± 1.37 ) ％,( 24.53 ± 2.28 ) ％ vs ( 2.17 ± 0.70 ) ％,P ＜ 0.01] and enhance the GJIC of PC-3 cells.EGCG of different doses could inhibit prostate cancer xenografted tumor growth,induce tumor cells apoptosis and inhibit angiogenesis.EGCG ( 20 and 40 mg/kg) could effectively up-regulate Cx43 mRNA expression in xenografted tumor (0.58 ± 0.08,0.80 ± 0.07 vs 0.42 ± 0.04,P ＜ 0.0 ).The effects had significant correlation with the dose-dependent of EGCG ( P ＜ 0.05 ).Conclusions EGCG could up-regulate the Cx43 expression and enhance the gap junction intercellular communication mediated by Cx43 in the prostate tumor,which provide the experimental evidence for the mechanism of its effectively inhibiting the prostate cancer growth.

Female ; Hamartoma ; pathology ; Humans ; Middle Aged ; Nasal Cavity ; pathology ; Nose Diseases ; pathology

Objective： To investigate the effect of c ytokines (IFN-γ，TNF and IL-1) on the expression of CD40 and CD40 ligand (CD4 0L) in monocytes/macrophages. Methods： The mRNA expression of C D40 and CD40L was measured by RT-PCR and the CD40，CD40L expression on the mono cytes/macrophages were detected by flow cytometric analysis. Results： IFN-γ，TNF and IL-1 could not only significantly up-regulate the mRNA levels of CD40 and CD40L in cultured monocytes/macrophages, but also increase t he expression of CD40 and CD40L. Antioxidant VitE could reduce the expression o f CD40 and CD40L induced by IFN-γ，TNF and IL-1. Conclusion： IFN-γ，TNF and IL-1 can stimulate high expression of CD40 and CD40L . Antio xidant VitE can partially inhibit the expression of CD40 and CD40L induced by cy tokines in cultured monocytes/macrophages.

Objective To determine the expression of interleukin-21 (IL-21) in patients infected with hepatitis B virus (HBV) and its association with HBV-DNA and ALT. Methods Clinical dates and blood specimen were collected from 25 unrelated healthy controls (HC) and 101 independent chronic HBV infected patients, including 25 patients in immune tolerant phase (IT), 25 in immune clearance phase (IC), 26 patients in inactive HBV carrier state (IA) and 25 patients in immune reactive phase (IR). Serum IL-21 levels were measured by Cytometric Bead Array (CBA). IL-21 mRNA and IL-21 receptor mRNA were measured by real-time PCR. Results Chronic HBV-infected patients had higher levels of serum IL-21 and IL-21 mRNA , with P <0.001 for both. In subgroup analysis, both serum IL-21 and IL-21 mRNA levels in IC, IR were higher than those in IT, IA and HC (all P < 0.001). Serum IL-21 level in IA was higher than that in HC and IT (P <0.001, P = 0.036). IL-21R mRNA levels were different between groups. Serum IL-21 level was associated with HBV-DNA (r = -0.472, P < 0.001), but not with ALT. Conclusion IL-21, up-regulated in chronic HBV infection, is associated with immune activity and may play a key role in HBV control.

Objective To investigate the effects of danhong injection on the apoptosis of SMMC-7721 heptoma cells.Methods The hepatocellular carcinoma cell line SMMC-7721 was incubated with different concentrations(0,2,4,8 μL·mL-1) of danhong injection, and cell morphology was studied under the microscope. The apoptosis index was detected by flow cytometry at 0,6,12,24 h after cultured with 2 μL·mL-1danhong injection. Cell proliferation was measured by MTT assay;Gene expressions of p53,Bax and Bcl-2 were detected by RT-PCR at different drug concentrations. Results The morphology of the hepatoma cells changed obviously,and the apoptosis index increased by danhong injection in a dose-dependant manner. The danhong injection decreased gene expressions of Bcl-2,and obviously increased p53 and Bax. Conclusion Danhong injection can dose-and time-dependently promote apoptosis of SMMC-7721 cells.

OBJECTIVETo compare the difference in clinical efficacy on polycystic ovary syndrome (PCOS) between electroacupuncture (EA) and dyne-35 and to explore the effect mechanism.

METHODSSixty-five patients were randomized into an EA group (33 cases) and a western medication group (32 cases). In the EA group, the selected acupoints were Danzhong (CV 17), Qimen (LR 14), Zhongwan (CV 12), Tianshu (ST 25), Guanyuan (CV 4), Zigong (EX-CA 1), Sanyinjiao (SP 6), Zusanli (ST 36) and Taichong (LR 3), etc. After the arrival of qi, electric stimulation was attached to the acupoints for 30 min. The treatment was given 3 times a week. In the western medication group, dyne-35 was prescribed on the 5th day of natural menstruation or withdrawal bleeding, one tablet a day, continuously for 21 days. The treatment cycle was 3 months in the two groups. The menstrual condition, body mass, body mass index (BMI), serum testosterone (T), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and LH/FSH were compared before and after treatment in the two groups. The clinical efficacy was assessed in the two groups.

RESULTSThe total effective rate was 90.6% (29/32) in the EA group and was 93.3% (28/30) in the western medication group. The efficacy was similar in the two groups (P > 0.05). After treatment, the levels of LH and LH/FSH were all reduced significantly in the two groups (all P < 0.01). After treatment, T level in serum was reduced apparently in the western medication group (P < 0.05). Before and after treatment, the differences in body mass and BMI in the EA group were more significant than those in the western medication group (P < 0.01, P < 0.05).

CONCLUSIONEA is the effective method for PCOS, similar to that of dyne-35. The effect of it for weight loss is superior to dyne-35 and no apparent adverse reactions happen. The effect mechanism of EA is related to the regulation of serum sexual hormone levels and their ratio, as well as to the regulation of body lipid metabolism.

Acupuncture Points ; Adolescent ; Adult ; Cyproterone Acetate ; administration & dosage ; Drug Combinations ; Electroacupuncture ; Ethinyl Estradiol ; administration & dosage ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Polycystic Ovary Syndrome ; blood ; drug therapy ; therapy ; Young Adult