Objectives　To evaluate prospectively the bone markers of serum osteocalcin (BGP),bone-specific alkaline phosphatase (BAP),C-terminal peptide of type I collagen (CICP),urinary pyridinoline (PYD),urinary calcium to creatinine value (uCa/Cr),and hydroxyprolin to creatinine ratio (uHOP/Cr),and to assess their responsiveness to subtle changes of bone metabolism,and furthermore to compare which of the bone markers are more sensitive to estrogen shortage caused by menopause.Methods　Markers of above bone formation and bone resorption were measured in 146 women classified into three groups: group I: 51 normal premenopausal women,group II:42 postmenopausal women,and group III: 53 bilateral ovariectimized women.Correlation analysis was made between these indexes and bone mineral density (BMD).Results　The mean values of the four bone markers of BGP,BAP,CICP,PYD in group II and III were significantly higher than those in group I (P<0.05).All correlations between the biochemical markers of BGP,BAP,CICP and PYD for bone turnover were highly significant and exhibited a significant connection to BMD.The best correlation was found between BGP and PYD,BGP and BAP.However,the levels of Ca/Cr and HOP/Cr made no difference in the three groups,and there was no correlation with any other indexes.Conclusion　In postmenopausal women (natural menopause and surgical menopause) suffering from severe E2 dificiency,BGP,BAP,CICP and PYD are significantly increased,indicating a clear correlation between E2 diffciency and these markers of bone turnover,so they are clinically useful in the assessment of bone turnover during changes of the estrogen status that modify the metabolic activity of the skeleton.But the traditional markers of Ca/Cr and HOP/Cr have limitations in gynecological practice because of lack of sensitivity and specificity.

Objective:To investigate the anti-hCG antibody level in vaginal lavage fluid after vaginal mucosa immunization with the recombinant lactobacillus excreting the human chorionic gonadotropin beta-subunit, hCG?.Methods:The recombinant Lb. casei hCG? was passaged in MRS containing no Erythromycin for 50 generations to get continual secretion. Radio immunol assay(RIA) was used to determine the level of hCG? in the supernatant. Mice of 8 weeks old were immunized with the recombinant continual expressing strain through vaginal inoculation. Two weeks later, level of the hCG? and anti-hCG? IgA antibody in the vaginal lavage was determined by radio-immunology assay and indirect ELISA respectively.Results:The hCG? was secreted by Lb. casei continually. The recombinant hCG? and anti-hCG? IgA antibody in the vaginal lavage were higher than the negative control group(P

Objective:To probe into if human molecular adjuvant hC3d3 promote the immunogenicity of hCG? antigen to human immuno-competent cells on the basis of fusion protein.Methods:The isolated B cells, the combination of B cells and T cells, PBMC and Raji cells were treated in vitro respectively with 1, 10 and 100 nmol/L hCG?, hCG?-hC3d3 or PWM for 8-12 days. The cell proliferation was determined by incorporation of [3H] thymidine. The Ig levels in the 12-day culture supernatants were measured by indirect ELISA. The Ig-secreting cells in the 10-day cultured lymphocytes were detected by the enzyme-linked immunospot(ELISPOT) assay.Results:It was found that the proliferation of B cells, the combined B and T cells, PBMC and Raji following exposure to hCG?-C3d3 fusion protein was significantly higher than that of hCG? alone. The levels of total Ig the in 12-day culture supernatants of B cell, the combined B and T cells, and PBMC treated with 100 nmol/L hCG?-C3d3 fusion protein were 4-fold, 10-fold and 10.85-fold more than that of hCG? alone. The Ig-secreting cells were significantly increased after treated with hCG?-C3d3 fusion protein compared to the hCG? alone.Conclusion:The human molecular adjuvant hC3d3 improves the immunogenicity of hCG? in human immuno-competent cells if fused to the antigen.

Objective:Investigating the expression of HIV-1 co-receptors CXCR4,CCR5 and chemoking SDF-1 in human placentae and trophoblasts is to explore the mechanism of in-utero transmission of human immunodeficiency virus type 1(HIV-1).Methods:Using semi-quantitative reverse transcriptase-polymerase chain reaction,detected the transcripts of CXCR4,CCR5 in placenta tissues and trophoblasts.Immunocytochemistry and immunohistochemistry analysis revealed the expression of CXCR4,CCR5 in primary cultured first trimester trophoblasts and villous tissue.Also the expression of SDF-1 in villi of first trimester,and the presence of SDF-1 in the culture of isolated trophoblasts were examined by in situ hybridization,immunohistochemistry and enzyme-linked immunosorbent assay.Results:CXCR4 and CCR5 mRNA were detected in placentae of various gestational phases and in first trimester trophoblasts.However,primary cultured trophoblasts were found to be reactive only with antibodies against CXCR4.In the villous tissue sections,CXCR4 was found in trophoblasts,while CCR5 in stromal cell and/or Hofbauer cell.First trimester trophoblasts expressed SDF-1 strongly and secreted SDF-1 in vitro.Conclusion:CXCR4 and CCR5 were expressed in placentae.Hence,they maybe involved in in-utero transmission of HIV-1 as co-receptors.However,SDF-1 expressed in trophoblasts may protect fetus from being invaded by X4-HIV-1.R5-HIV-1 may infect CCR5+ stromal cells and/or Hofbauer cells through placental disruptions.

AIM: To study the effect of activated protein C(APC) at different concentrations on apoptosis of human umbilical vein endothelial cells(HUVECs) induced by lipopolysaccharide(LPS).METHODS: The HUVECs were induced by LPS(1.0 mg/L) as apoptotic model that was administered by different concentration of APC(10 ?g/L or 50 ?g/L).Meanwhile,the control group and induced apoptosis group induced by LPS(1.0 mg/L) stimulation were also set up.The changes of cellular ultrastructures were observed under electron microscope.The DNA ladder and TUNEL fluorescent staining were measured in cells.Annexin-Ⅴ/PI double staining was used to measure the cell apoptosis rate by flow cytometry.Cell survival rate was measured by MTT assay.The proliferating cell nuclear antigen(PCNA) expression levels in cells were also measured by Western blotting to reflect the proliferation of the cells.RESULTS: There were significant apoptotic changes in the cells induced by LPS,but the apoptotic changes were reduced and apoptosis rates were decreased in the cells treated with APC.Meanwhile,cell survival rate and the protein levels of PCNA were increased after APC treatment,particularly at the concentration of 50 ?g/L,which showed difference when compared with those induced apoptosis group by LPS(P

Objective To investigate the role of connexin 43(Cx43)in the apoptosis of pancreatic cancer cell line BxPC and its possible mechanism.Methods pcDNA-Cx43,pcDNA-Cx43N,pcDNA3.0,siRNA-Cx43 and siRNA-NC were transfected into BxPC3 cells via liposome method.Cx43 protein and Cytochrome C(Cyt C)concentration was determined by Western blot,and the apoptosis was analyzed by Annexin V/PI binding assay.The mitechondria apoptosis pathway involved in Cx43 associated apoptosis was examined which contains the depolarization of mitechondrial membrane potential (MMP);fluorospectrophotometer was used to measure the activities of caspase-3 and caspase-9. Gap junction intercellular communication(GJIC) was determined by dye-transfer method.Results Cx43 protein expression increased after BxPC3 transfeetion,apoptosis rate increased from(6.35±0.43)%in empty vector transfection group to(14.29±1.24)%;after H202 treatment,apoptosis rate increased from(20.34±2.47)%to(31.27±2.56)%(P<0.05).Meanwhile,mitochondrial membrane potential was decreased,Cyt C was increasingly released from mitochondria,caspases activities were increased;after siRNA43 interference,apoptosis rate decreased from(7.42±0.47)% to(5.19±1.37)%,after H_2O_2 treatment,apoptosis rate decreased from (19.43±1.71)%to(11.67±1.97)%(P<0.05).Decreased mitochondrial membrane potential and Cyt C release were observed,caspases activities were decreased.GJIC of pcDNA-Cx 43 transfection group increased from 14.52±0.57 to 23.05±3.84.and it increased from 1.70 ±0.24 to 3.84 ±0.45 in the presence of β-GA(P<0.05).But the apoptosis rate was not significantly different.Conclusions Cx43 could promote BxPC3 apoptosis via mitochondrial apoptotic signal pathway,and the possible mechanism included signal pathway other than GJIC.

Objective To investigate the relationship between the activity of Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway in cerulein-induced pancreatic acinar cell line AR42J. Methods The in vivo model of AP was induced by cerulean treated pancreatic acinar cell line AR42J, then RPM and AG490 were given for intervention. Western blot was used to determine theexpressions of JAK1 and phosphorylation JAK1 ( P JAK1 ) , STAT1, PSTAT1 and TNFα, IL-1β, IL-6. The expressions of IL-6, IL-1 β, and TNFα mRNA were measured by RT-PCR. Survival rate of cells was evaluated by trypan blue stain. Results The relative expressions of JAK1, P JAK1, STAT1, P STAT1 and TNF-o, IL-1β, IL 6 without cerulean treatment were 0.09 ±0.04,0.14 ±0.08,0.21 ±0.09,0.12 ±0.12,0.10 ±0.02,0.08 ± 0.03,0.02 ± 0.02. After cerulean treatment, the expressions of abovementioned protein increased in a time-dependant manner, the expressions at 24h were 0.53 ± 0.09,0.53 ± 0.13,0.56 ± 0.09,0.55 ± 0.10,0.25 ± 0.04,0.25 ±0.09,0.27 ±0.07, which were significantly higher than those in the control group (P ＜0.05). 2 4 h after RPM and AG 4 9 0 inhibition, the expressions of TNF-α, IL-1 β, IL-6 proteins significantly decreased to 0.17 ± 0.03and 0.17 ± 0.01,0.15 ± 0.05 and 0.14 ± 0.07,0.19 ± 0.04 and 0.19 ± 0.05; their expressions of mRNA significantly decreased ( P ＜ 0.05 ). The cell survival rates in RPM and AG490 treatment group were (72.4 ± 11.2) %, (69.7 ± 9.8 ) %, and in cerulein-stimulated cells (42.2 ± 12.3 ) % ( P ＜ 0.05 ).Conclusions The JAK1/STAT1 signaling pathway was involved in pancreatic inflammatory response with cerulein stimulation. Early treatment with inhibitors to the JAK1/STAT1 signaling pathway might control the inflammatory response in acute pancreatitis.

Objective To study the protective effects of activated protein C (AFC) on the apoptosis of endothelial cells induced by lipopolysaccharide (LPS) in order to clarify the mechanisms associated with the expression of some genes related to apoptosis. Method The human umbilical vein endothelial cells were incubated with LPS (1.0 μg/mL) for one hour to make the models of cell apoptosis, and then the different concentrations of AFC (10 ng/mL and 50 ng/mL) were added to the models of cell apoptosis as treatment group. Therefore, there were two groups, model group and APC treated group. The factors related with apoptosis such as P53, Bax, Bcl-2, and caspase-3 mRNA or protein level were measured by using RT-PCR and Western blotting. Results Compared with LPS stimulated cells, the expressions of P53, Bax and caspase-3 mRNA and levels of protein were decreased and the expression of Bcl-2 mRNA and protein level were increased in APC treated cells particularly in APC 50 ng/mL treated cells (P <0.05). Conclusions The APC inhibits the apoptosis of HUVECs induced by LPS via regulating the mitochondrial-dependent apoptosis pathway, and it may become a novel therapeutic agent for infection disease.

The reform of performance appraisal and salary system is key to public hospitals reform.This paper introduced practices of the factor performance and salary system reform based on the Balance Scorecard in hospitals.It established a four-dimension performance appraisal structure,a motive salary allocation model covering all members of a hospital.This would help build the performance appraisal and salary allocation system meeting present needs of public hospitals in China.

Objective To investigate the effects of ginsenoside Rg3 on growth and apoptosis of gastric cancer cell line MKN-45 and SGC-7901 in vitro. Methods MKN-45 and SGC-7901 cells at logarithmic growth phase were obtained, and were cultured with ginsenoside Rg3 of different concentrations (20, 30, 40, 50 μg/mL) for 24, 48 h or 24, 48 and 72 h. Cells cultured without ginsenoside Rg3 were served as controls. The inhibition rates of ginsenoside Rg3 on MKN-45 and SGC-7901 cells were detected by MTT assay, apoptosis rate of SGC-7901 cells was determined by Annexin V/PI double staining flow cytometry, cell cycles of SGC-7901 cells were analysed by flow cytometry, and morphological changes of SGC-7901 cells in 50 μg/mL ginsenoside Rg3 treatment group were observed by transmission electron microscopy. Results The inhibition rates on MKN-45 and SGC-7901 cells in each ginsenoside Rg3 treatment group were significantly higher than those in control group (P < 0.05), and the inhibition rates increased with the concentrations of ginsenoside Rg3 and time of culture ( P < 0.05). Compared with control group, the apoptosis rates of SGC-7901 cells and percentages of cells in G_1/G_1 cell cycle in each ginsenoside Rg3 treatment group were significantly increased in a concentration and time dependent manner. Typical morphology of SGC-7901 cell apoptosis was observed by transmission electron microscopy in 50 μg/mL ginsenoside Rg3 treatment group. Conclusion Ginsenoside Rg3 has significant inhibition effect on gastric cancer cell lines in vitro with a concentration and time dependent manner, the mechanism of which may involve the induction of gastric cell line apoptosis.