Objective To investigate the effects of integrin β1 gene expression inhibited by short hairpin RNA (shRNA) on invasion of pancreatic carcinoma PANC1 cells in vitro,and investigate the mechanism.Methods The eukaryotic expression plasmid of shRNA targeting integrin β1 gene ( integrin β1 shRNA) and control eukaryotic expression plasmid shRNA (c-shRNA) was constructed and was transfected into PANC1 cells.The cells without plasmid transfection were used as control.The expressions of integrinβ1,MMP 2,MMP 9 mRNA and protein were detected by real-time PCR and Western blotting.The invasive ability of PANC1 cells was observed with Transwell cell culture chamber.Results Integrinβ1 mRNA expressions in integrinβ1 shRNA group,c-shRNA group and control group were 0.0029 ± 0.0004,0.0131 ± 0.0009,0.0138 ± 0.0005 ; the expressions of integrinβ1 protein were 0.0159 ± 0.0062,0.3215 ± 0.0126,0.3107 ±0.0094; the inhibitory rate of integrinβ1 mRNA and protein expression in integrinβ1 shRNA group was (78.6 ±7.2 ) ％ and (92.9 ± 3.2) ％ ( P ＜ 0.01 ).But there was no difference between the c-shRNA group and control group (P =0.2999).Number of penetrating cells in integrinβ1 shRNA group decreased from 52 ±5 to 21 ±4( P ＜ 0.01 ) ; the expression of MMP 2 and MMP 9 mRNA decreased from 0.592 ± 0.073,0.847 ± 0.069 to 0.102 ± 0.034,0.273 ± 0.071 ; the expression of M MP2 and MMP 9 protein decreased from 0.225 ± 0.046,0.416 ±0.081 to 0.059 ±0.013,0.106 ±0.022(P ＜0.05).Conclusions Recombinant integrinβ1 shRNA expression plasmid can effectively inhibit the expression of integrinβ1 gene and suppress the invasion of PANC1 cells in vitro by down-regulating MMP 2 and MMP 9 gene expression.

Objective To summarize the epidemiological feature of plague cases oceuwed in China.Methods The epidemiological and clinical data from 1981 to 2006 year in China were analyzed with descriptive study method.Result Nine hundred and seveneteen human plague cases were diagnosed in 9 provinces(regions) from 1981 to 2006 years,105 cases died,the mortality rate being 11.45%,and they distributed in 69 counties (cities or banners).In Qinghai Province 108 cases were diagnosed,the mortality rate was 46.30%(50/108),the cases distributed in 17 counties(cities);137 cans in Guizhou,distributing in 2 counties(cities);517 cases in Yunnan,distributing in 26 counties(cities).Plague cases peaked separately in 1983,1990,1996 and 2000 years,they were 25,75,98 and 254 separately.The principal spreading ways were breathing flying particles,touching,skinning and eating marmot in Qinghai;750 cases were of bubonic plague,among whom 4 cases in Tibet died,the fatality rate was 0.53(4/750);121 cases were of pneumonic prague,among whom 65 cases died,was accounting for 53.72%(65/121);31 cases were of septieaemic plague,and 30 cases died(one cases was cured in Inner Mongolia),accounting for 96.77%(30/31).Others were brain plague,intestinal plague,tonsil plague and plague cellulites,which were cured.Conclusion From 1990,human plague epidemical scope and intensity is enlarging continuously compared with 1980-1990 and there is a trend of going up gradually in China.

Objective:To investigate the efficiency of carbon nanotube(CNT)-PAMAM mediated entrance of anti-survivin oligonucleotide into HepG2 cells,and its effects on the proliferation of HepG2 cells.Methods:CNT-PAMAM-anti-survivin oligonucleotide compounds were prepared and characterized by AFM and 1% agarose gel electrophoresis analysis.TEM was used to observe the distribution of CNT-PAMAM-ASODN compounds in HepG2 cells.CNT-PAMAM-ASODN compounds were added into the medium and co-cultured with HepG2 cells for 24 h,48 h,72 h,and 96 h at 37℃,5% CO_2.MTT method was used to detect the effects of ASODN and CNT-PAMAM-ASODN on the proliferation of HepG2 cells.Results:CNT-PAMAM-ASODN compounds were successfully synthesized via AFM and agarose gel electrophoresis.TEM showed that the compounds were located in the cytoplasm.When CNT-PAMAM-ASODN(1.0 ?mol/L)and ASODN(1.0 ?mol/L)were used for a 48 h culture,the inhibitory rates of HepG2 cells were(45.97?4.28)% for CNT-PAMAM-ASODN compounds group,(9.33?0.85)% for ASODN group,and(6.37?0.69)% for CNT-PAMAM group.CNT-PAMAM-ASODN compounds at 1.5 ?mol/L inhibited HepG2 cells by(70.22?7.25)%,and the inhibitory effects were in a time-and concentration-dependent manner.There was statistical difference between experiment group and control group(P

COP9 Signalosome Complex ; Carcinoma, Hepatocellular ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; chemistry ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Gene Expression Regulation, Neoplastic ; Humans ; Intracellular Signaling Peptides and Proteins ; chemistry ; genetics ; metabolism ; Peptide Hydrolases ; chemistry ; genetics ; metabolism ; Phosphorylation ; Phosphoserine ; metabolism

Objective:To investigate the effect of blocking BRCAA1 gene expression with siRNA on the proliferation of tumor cell line MCF-7 and Rb gene expression.Methods:RNAi was employed to specifically knock down BRCAA1 expression.MCF-7 cells were transfected with complexes constructed with lipids and chemically synthesized Pre-designed anti-BRCAA1 siRNAs.The total RNA was isolated and reversely transcribed after 48 h.The expressions of BRCAA1 and Rb mRNA were determined by Real-Time PCR.Results:Compared with negative control,transfected MCF-7 cells had a 42.3% decrease in expression of BRCAA1 mRNA and an 11.1% increase in Rb mRNA expression.The inhibitory rate of MCF-7 cells proliferation was(81.6?6.1)%.Conclusion:There may be some antagonistic effect between BRCAA1 gene and Rb gene in proliferation of tumor cells,which provides a potential target for anti-tumor gene therapy.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Staging ; Phosphorylation ; S-Phase Kinase-Associated Proteins ; metabolism

OBJECTIVETo better understand the molecular mechanism of tumorigenesis and progression, the monoclonal antibody against G3BP (Ras-GAP SH3 binding protein), which serves as an important downstream effector of Ras signaling, was prepared, characterized and utilized in analysis of various human tumors.

METHODSBy using the prokaryotic expression vector pGEX-5X1, GST-G3BP fusion protein was expressed in E. coli BL21 under induction of IPTG. Purified GST-G3BP fusion protein was used to immunize BALB/c mice. The monoclonal antibody against G3BP was produced through conventional hybridoma method and characterized by ELISA, Western blot and immunohistochemical staining.

RESULTSA hybridoma cell line secreting anti-G3BP IgG1 subtype antibody was obtained. Western blot and competitive inhibition assay showed that the antibody was G3BP-specific. Immunohistochemical staining demonstrated that G3BP was over-expressed in formalin-fixed and paraffin-embedded tissues of some human tumors, such as lung cancer, colon cancer, gastric cancer and breast cancer. In breast cancer specimens, the degree of G3BP expression correlated positively with the presence of lymph node metastasis and c-erbB2 expression.

CONCLUSIONSThe G3BP-specific monoclonal antibody derived from recombination protein can be used in ELISA, Western blot and immunohistochemical assay. It may provide an important tool in analysis of G3BP in in vitro and in vivo experiments. Besides, G3BP may serve as another prognostic marker for breast cancer.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Biomarkers, Tumor ; Breast Neoplasms ; metabolism ; pathology ; Carrier Proteins ; genetics ; immunology ; metabolism ; DNA Helicases ; Female ; Genetic Vectors ; Humans ; Hybridomas ; secretion ; Lymphatic Metastasis ; Male ; Mice ; Mice, Inbred BALB C ; Poly-ADP-Ribose Binding Proteins ; RNA Helicases ; RNA Recognition Motif Proteins ; Receptor, ErbB-2 ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Tumor Cells, Cultured

Objective To investigate the effect of 4-hydroxytamoxifen on the expression of pituitary tumor transforming gene (PTTG) in GH3 prolactinoma cells. Methods RT-PCR and Western blotting were employed to detect the expressions of PTTG mRNA and protein in human GH3 prolaetinoma cells. Different concentrations of estradiol (E2) or 4-hydroxytamoxifen (OHTam) were addedl into the hormone-depleted medium, and the viable cell number and expression levels of PTTG mRNA and protein were measured. Results The growth of OH3 prolaetinoma cells was significantly inhibited in hormone-depleted medium. E2 at a concentration of 1×10<'-8> mol/L obviously promoted the cell growth, the effect of which was inhibited by the application of OHTam (1×10<'-6> mol/L) to cause slowed cell growth. The expressions of PTTG at both the mRNA and protein levels were detected in detected in untreated GH3 prolactinoma cells, and OHTam at the concentration of 1×10<'-6> mol/L significantly inhibited their expressions. Conclusion The growth of GH3 cells is estrogen-dependant and can be inhibited by estrogen antagonist OHTam, which also results in reduced expression of PTTG gene in the cells.

Objective To explore the effect of Objective Management in humanity qualities training for practice nurse.Methods Student nurses practiced in our hospital in 2009 served as a control group (n=186) and those in 2010 served as experimental group (n=200). The control group were evaluated by traditional method, and the experimental group were managed by Objective Management in training of humanity qualities. The patient satisfaction, self-satisfaction and teacher satisfaction of the two groups were compared.Results There were no significantly statistical differences of the patient satisfaction, self-satisfaction and teacher satisfaction between the two groups in pre-practice-time(P>0.05), while there were significant differences in the post-practice-time(P＜0.05).Conclusions Results suggest that Objective Management can improve the humanity qualities of the practice nurse. It is worth spreading.

OBJECTIVETo investigate the expression of PDGF receptor-beta and its correlation with extracellular matrix in hepatic tissue during hepatic fibrosis.

METHODSThe model of hepatic fibrosis in rats was induced by carbon tetrachloride. PDGF receptor-beta subunit, collagen I, collagen III and a-SMA in hepatic tissues of these rats were examined using immunohistochemistry. The correlation between PDGF receptor-beta subunit and collagen I, III was analyzed using SAS software after the results of immunohistochemistry were semi-quantified.

RESULTSPDGF receptor-beta subunit and a-SMA were not detected in normal controls. Collagen I and III were distributed in the portal tracts and beneath the endothelia of the central veins and of the Disse spaces. Two weeks after CCl4 injection, the PDGF receptor-beta and a-SMA were detected, and the expression of collagen I and III increased. At the end of 4 and 6 weeks, the above four proteins were further increased. Two weeks after CCl4 injection, PDGF receptor-beta had no apparent correlation with collagen I and III. However, PDGF receptor-beta had a significant correlation with collagen I and III 2 weeks later, and the correlation coefficient was 0.74 and 0.60 respectively at 4 weeks, and 0.83 and 0.67 respectively at 6 weeks. PDGF receptor-beta had a significant correlation with a-SMA during the whole process of hepatic fibrosis and the correlation coefficient was 0.62, 0.69 and 0.81, respectively at the time of 2, 4 and 6 weeks after CCl4 injection.

CONCLUSIONThe PDGF receptor-beta was overexpressed during the process of hepatic fibrosis development, and it significantly correlated with collagen I and collagen III.

Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Extracellular Matrix ; metabolism ; Liver ; metabolism ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptor, Platelet-Derived Growth Factor beta ; biosynthesis ; genetics