Adenocarcinoma ; diagnosis ; genetics ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; genetics ; Carcinoma, Squamous Cell ; diagnosis ; genetics ; Cyclin-Dependent Kinase Inhibitor p16 ; blood ; metabolism ; DNA Methylation ; Genes, p16 ; Humans ; Lung Neoplasms ; diagnosis ; genetics ; Tumor Suppressor Proteins ; blood ; metabolism

Objective To study the effect of mechanical stretch on the expression of human beta-defensin-3 (HBD-3) in alveolar epithelial cells(A549 cells) elicited by interferon-gamma(IFN-γ) and to investigate the role of HBD-3 in the pathogenesis of ventilator-associated pneumonia (VAP) . Method A549 cells cultured in vitro were treated with mechanical stretch (group S), 10 ng/ml IFN-γ (group I) ,and 10 ng/ml IFN-γ with mechanical stretch (group IS), respectively. Cells without treatment served as controls (group C). Cells were stretched by 20% amplitude of stretch at 30 cycles/mm by Flexercell-4000[TM]Unit for 2 h, 4 h, and 6 hours. The HBD-3 mRNA expression was determined by real-time RT-PCR after treatment. After 6 hours, treatment, cells were cultured for 24 hours and the expression of HBD-3 was examined by laser scanning confocal microscope. The experimental data were statistically analyzed by using one-way ANOVA analysis and q-test. Results The expression of HBD-3 mRNA in A549 cells could not significantly be changed by mechanical stretch alone. Compared with group C,the HBD-3 mRNA expression after treatment with 10 ng/ml IFN-γ for 2 hours,4 hours and 6 hours increased significantly by (2.63 C,the HBD-3 mRNA expression after treatment with 10 ng/ml IFN-γ and mechanical stretch for 2 hours,4 hours and 6 hours increased by (1.54 were significantly lower than those in group I (P < 0.01). The HBD-3 expression in group IS after mechanical stretch for 6 significantly different from than in group C. Conclusions Mechanical stretch can significantly suppress the up-regulation of HBD-3 in alveolar epithelial cells elicited by IFN-γ, and this may be one of the explaina-tions that patients under mechanical ventilatiori(MV) have a higher risk of VAP.

Objective Using the third high sensitive cardiac troponin I assay to establish the reference intervals of serum cardiac marker high sensitivity cardiac troponin I ( hs-cTnI ) in apparently healthy subjects of Beijing area, and to explore the relationship between cTnI and other parameters.Methods Using random selection method, a total of 440 serum samples were collected from apparently healthy subjects of Beijing area.Among them, 217 cases were male, average age was (50.0 ±14.3) years old;223 cases were female, average age was ( 49.0 ±14.0 ) years old.All of these serum samples were detected hs-cTnI using i2000 Abott biochemical analyzer and ARCHITECT STAT Abbott assay kit.And the 99th percentile upper reference limit was used as the reference interval.Non-parametric tests were used to compare the different groups, the U Mann-Whitney test was used between two groups, the Kruskal-Wallis test was used in many groups, and the correlation analysis was used Spearman method.Results The 99th percentiles of hs-cTnI in 440 apparently healthy subjects is 9.66 ng/L.While the male group is 9.66 ng/L, and the female group is 8.25 ng/L.The serum hs-cTnI level of male is higher than female ( 9.95 ng/L vs 8.25 ng/L, P<0.05 ).According to the spearman correlation analysis, the serum hs-cTnI level appears positive correlation with age, brain natriuretic peptide (BNP), alanine aminotransferase (ALT),aspartate transaminase ( AST) ,gamma-glutamyl transferase ( GGT) ,total bilirubin ( TBIL) ,serum creatinine ( SCR) , hemoglobin (HGB)(P<0.05,R>0),and appears negative correlation with platelet (PLT)(P<0.05,R<0).Conclusion The serum hs-cTnI level apparently differents with gender, so it is necessary to establish reference intervals among different gender group of Beijing area.

Objective To investigate the effect of amiloride pretreatment on the acute lung injury (ALI)induced by lipopolysaccharide (LPS) in rats. Methods Thirty-two pathogen-free male SD rats weighing 200-250 g were randomly divided into 4 groups (n = 8 each); group Ⅰ received iv normal saline (group C); group Ⅱ ALI received iv LPS 6 mg/kg (group ALI); group Ⅲ received iv amiloride 10 mg/kg (group A) and group Ⅳ received amiloride 10 mg/kg iv 30 min before iv LPS ( group AL). The animals were killed by exsanguination at 6 h after iv LPS infusion. The lungs were immediately removed. Microscopic examination of lung tissue was performed. The left lung was lavaged. The total protein (TP), TNF-α and macrophage inflammatory protein-2 (MIP-2)concentrations in broncho-alveolar lavage fluid (BALF) were measured. The W/D weight ratio and the myeloperoxidase (MPO) activity and expression of Na-H exchanger-1 ( NHE1 ), p38MAPK and extracellular signal-regulated kinase (ERK) in lung tissue were determined. Results LPS significantly increased ALI score (0 = slightest, 4 = severest), W/D lung weight ratio, TP, TNF-α and MIP-2 concentrations in BALF and MPO activity and the expression of NHE1, p38MAPK and ERK in the lung as compared with. control group. Amiloride pretreatment significantly attenuated LPS-induced changes except p38MAPK expression. Conclusion Pretreatment with amiloride can attenuate LPS-induced ALI by inhibition of ERK activation.

Air ; analysis ; Hydrogen Peroxide ; analysis ; Spectrophotometry ; methods ; Workplace

Objective　To clone the partial positive regulatory fragment of Na+/H+ exchanger-1 (NHE-1) gene from human lung cancer cells. Methods　After BamHⅠ and EcoRⅠ cut sites were added to the 5' ends of the upstream and downstream primers respectively, the partial positive regulatory sequence of NHE-1 gene was cloned with the length of 170 bp from genomic DNA of lung cancer cell line A549 cells with PCR method. The cloned fragment was ligated to plasmid pUC18. Finally, the constructed recombinant was identified with enzyme cut, PCR and DNA sequencing. Results　The cloned fragment was about 170 bp in size and successfully ligated to pUC18 with identifiation of double enzyme cut and PCR. DNA sequencing approved that the fragment cloned was objective one with 168 bp in length. Compared with the reported sequence, two t were lost. Conclusion　The positive regulatory fragment of NHE-1 gene from human lung cancer cells was successfully cloned.

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

Adolescent ; Adult ; Aged ; Biopsy ; Delayed Graft Function ; diagnosis ; pathology ; Female ; Graft Rejection ; diagnosis ; pathology ; Humans ; Kidney ; pathology ; physiopathology ; Kidney Function Tests ; Kidney Transplantation ; pathology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo understand the demography changes and behaviors in drug users.

METHODSelf-reported questionnaires was used and longitudinal investigation was conducted in one of the detoxication centers in Beijing in 1998 and 2000. Drug users were randomly chosen.

RESULTSResults showed that age of drug users tend to become younger with the numbers of drug users aged below 25, increased from 18.7% in 1998 to 28.2% in 2000. Majority of drug users remained males, but the proportion of females seemed to increase. Distribution of occupation showed that the largest increase fell among individual enterprisers, from 15.2% in 1998 to 25.9% in 2000. With educational back-ground, the proportion of lower than elementary education level, including illiterate, increased. Fifty percent of drug users were unmarried which increased from 40.8% in 1998 to 53.2% in 2000. Needle sharing was quite common, 16.5% in 1998 and 11.9% in 2000, but the decrease was not statistically significant (P > 0.05). Fifty-seven point three percent of the injecting drug users did not have constant partners to share equipments. Proportion of extra-marriage sexual practice increased from 12.5% in 1998 to 27.5% in 2000, and significant difference (Chi-square = 12.50, P < 0.001). Multiple partners in extra-married drug users was also found (mean = 2). Compared to 1998, condom use during every sexual practice increased in 2000, but 47.7% drug users still never used condom.

CONCLUSIONIn summary, as the quick increase of drug users, sharing of injecting equipment and high-risk sexual behavior, including multiple partners and unprotected sex, were quite common, with the possibility of HIV epidemic in drug users.

Adult ; China ; epidemiology ; Demography ; Female ; HIV Infections ; epidemiology ; Humans ; Longitudinal Studies ; Male ; Needle Sharing ; adverse effects ; Prevalence ; Risk Factors ; Safe Sex ; statistics & numerical data ; Substance Abuse, Intravenous ; epidemiology ; Surveys and Questionnaires

Objective To investigate the relationship between Wnt5a gene and E-cadherin or vimentin gene in breast cancer cell line MCF-7. Methods RT-PCR was used to detect the mRNA expression of Wnt5a, E-cadherin and vimentin in breast cancer MCF-7 cells and the normal human mammary epithelial cell line MCF-10A, respectively, and their correlation was analyzed. Results The mRNA expression levels of Wnt5a and E-cadherin in cell line MCF-7 were significantly lower than those in cell line MCF-10A [(16.93± 2.97)%vs. (27.47±2.76) %, (12.97±1.35) % vs. (20.43±2.60) %, both P<0.05]. The mRNA expression level of vimentin in cell line MCF-7 was significantly higher than that in cell line MCF-10A [(16.53±0.85)%(6.33± 2.08) %, P<0.05 ]. In cell line MCF-7, the expression of Wnt5a was positively related to E-cadherin (г=0.997, P<0.05), but it was negatively related to vimentin (г=-0.998, P<0.05). Conclusions The expression of Wnt5a in human breast cancer cell line MCF-7 is significantly lower than that in cell line MCF-10A, which indicates that Wnt5a is a cancer suppressor gene in breast cancer. The expression of Wnt5a in cell line MCF-7 is positively related with E-cadherin, and it is negatively related with vimentin. Wnt5a may cause invasion and metastasis of breast cancer cell through the breast epithelial mesenchymal transitions.