Objective To assess the safety and efficiency of Angioseal device in patients undergoing coronary percutaneous procedure.Methods A prospective trial was carried out in 260 patients undergoing angiograph and angioplasty during october 2002 to July 2003.All patients were divided into two groups: Angioseal closure device and manual compression.Results In angiography,the time to hemostasis was (1.8?0.9)min by Angioseal and (25.3?13.4)min by manual compression(P

Objective:Toprepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods: The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP-1 in biodegradable PELA. The diameter of the microsphere, quantity of virus encapsulated, loading rate, and releasing kinetics were measured. HepG2 cells were infected with the microspheres; the infection efficiency was examined by fluorescent microscope; and the ultrastructure was observed by TEM. The expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR, and the proliferation of HepG2 cells was detected by MTT assay. Results: The microsphere encapsulating recombinant TIMP-1 adenovirus was successfully constructed, with its diameter, entrapment efficiency, and virus loading rate being 1.965, 60.0%, and 10.5×10~8/mg, respectively. About 60% of the viruses were released within 120 h, and the total releasing time was longer than 240 h. Infection with rAdTIMP-1 PELA microsphere efficiently induced TIMP-1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion: PELA microsphere encapsulating recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.

Objective To investigate the effects of the interaction between environmental factors and monoamine oxidase A (MAOA) gene polymorphism in Han and Uygur on alcoholics in Xinjiang. Methods The data of childhood abuse and domestic aggressive behaviors were collected from 284 patients with alcohol dependence from Xinjiang using self-made questionnaires, Modified Overt Aggression Scale (MOAS) and Barratt Impulsivity Scale-11 (BIS-11). The meth?ods of PCR and DNA sequencing technique were conducted to detect rs1137070 single nucleotide polymorphism loci of MAOA gene. Logistic regression analysis was performed to adjust for interaction effects of gene and childhood abuse on domestic violence. Results The scale evaluation identified 143 patients with and 138 patients without domestic violence. Childhood abuse and gene were both risk factors in domestic violence. The interaction effect of childhood abuse with rs1137070 was significant. the relative excess risk, the interaction attribution ratio and the interaction index were 1.00,0.14 and 1.20, respectively. Conclusions The interactions between genes and environmental risk factors may contribute to the domestic violence in Han and Uygur on alcoholics in Xinjiang.

Objective To investigate the effect of phenylhexyle isothiocyanate (PHI) on demethylation and activation of transcription gene p15 in acute leukemia cell line Molt-4. Methods DNA sequencing and modified methylation specific PCR (MSP) were used to screen p15-M and p15-U mRNA after Moh-4 cells were treated with PHI. P15 mRNA was measured by RT-PCR. Pl5 protein was detected by Western blotting. Results Hypermethylation of gene pl5 was apparently attenuated and activation of transcription p15 gene was de novo after 5 days exposure to PHI. PHI enhanced both the expression of p15 mRNA and p15 protein in a concentration-dependent manner. The ratio of the gray scale of p15 mRNA strap was 0.17±0.12 in control, 0.29±0.14 in PHI 10 μmol/L, 0.55±0.07 in PHI 20 μmol/L, 0.93±0.13 in PHI 40 μmol/L. Conclusion PHI could active demethylation and transcription of gene p15.

BACKGROUND:As current studies on isolation, culture andcryopreservation of human amniotic epithelial cells (hAECs) are relatively scattered, it is difficult to form a comprehensive and effective solution to meet the clinical needs of stem cells for transplantation in future.OBJECTIVE:To establish the technology of isolation, culture and cryopreservation of hAECs, and to study the biological characteristics of hAECs.METHODS:Orthogonal method was used to study the effects of different factors on the separation, culture and cryopreservation, and range method was adopted to analyze the data to optimize the separation, culture and cryopreservation. We performed cell primary and passage cultures, morphology observed by microscope, drawn cell growth curve and flow cytometry assay, immunofluorescence staining, hepatocyte like cell differentiation to study the biological characteristics of hAECs.RESULTS AND CONCLUSION:(1) The optimal hAECs separation conditions were as follows:trypsin digestions were conducted at a concentration of 0.25%, four times, once for 20 minutes digestion; optimal conditions of culture were 4×108/L cell seeding density, 10 μg/L epidermal growth factor, 5% serum; optimal conditions of cryopreservation were 1×1010/L cell cryopreservation density, 10% dimethyl sulfoxide, 80% serum. (2) The primary cells were adhered to the wall in 2-3 days, exhibiting irregular polygon, paving stone-like growth. Cell adherence and growth rate were accelerated after subculture, and the growth and proliferation ability of passage 2 cells were not significantly decreased after cryopreservation and resuscitation. (3) Immunofluorescence staining showed that the primary cells strongly expressed SSEA-4 and CK19, but did not express Vimentin, CD45 and HLA-DR. The immunophenotype statistics of the primary and passage 4 cells showed the epithelial mesenchymal transition of hAECs in culture process. (4) Immunofluorescence staining showed that the liver cell marker expression of ALB, CK18 was significantly increased after hAECs were induced to differentiate into hepatocyte-like cells. Glycogen staining revealed glycogen synthesis in hAECs after 3 weeks of induction. To conclude, hAECs are easy to obtain and have strong proliferation ability in vitro, and express surface markers for undifferentiated embryonic stem cells.

Objective To investigate the effect of hydrogen peroxide (H_2O_2) on apoptosis and calcium ion concentration of skeletal muscle satellite cells (SMSCs) in rats, and to explore the protective effect of erythropoietin (EPO).Methods The cultured SMSCs were divided into five groups: control group,H_2O_2 group, 10, 20 and 40 U/ml EPO intervention groups.Apoptosis rates and calcium ion concentration of SMSCs were analyzed by flow cytometry, and the morphology of apoptotic cells was observed by Hoechst33258 staining.Results The apoptosis rates showed significant differences (all P<0.05) among (1.93±0.57)% in control group, (22.13±1.79)% in H_2O_2 group, (16.47±2.53)%, (4.97±0.55)% and (2.93±0.47)% in 10, 20 and 40 U/ml EPO intervention groups, respectively.And calcium ion concentrations in SMSCs were 12.67 ±0.32, 27.90±0.06 and 44.53±0.93 in 10, 20 and 40 U/ml EPO intervention groups, respectively.There was significant difference in calcium ion concentration between H_2O_2 group and control group (9.70±0.09 vs.51.37± 0.64, P< 0.05).Morphology of apoptosis was observed by Hoeehst33258 dye stains in 10, 20 U/ml EPO intervention group and H_2O_2 group, while there were less apoptotic bodies in 40 U/ml EPO intervention group and control group.Conclusions EPO might have protective effects on SMSCs injured by H_2O_2 through inhibiting apoptosis and calcium ion releasing from SMSCs.

Objective: To investigate the effects of silk fibroin on the immobilization of thrombin. Methods: The immobilized thrombin was prepared using silk fibroin as the carrier and glutaraldehyde as the crosslinking agent. With activity yield as the index, the process conditions of silk fibroin immobilized thrombin were determined by an orthogonal test. Results:The optimum process con-ditions of immobilized thrombin treated with silk fibroin were as follows:the immobilization time was 6 h, the enzyme dosage was 2 400 NIH·g-1 casein, the temperature was 25℃ and pH was 7. 6. The activity recovery of immobilized thrombin was 67. 22%. Conclu-sion:Silk fibroin has the positive immobilization effect on thrombin.

Objective To express the hemagglutinin of H7N9 in Pichia pastoris and analyze its immunogenicity. Methods The HA [1 -525 amino acids(aa)] of H7N9 [A/Hangzhou/1/2013(H7N9)] lacking the C-terminal transmembrane anchor-coding sequence was amplified by PCR and cloned into the expression vector pPICZαA.The plasmid pPICZ-HA7-S-was transformed into P.pastoris and the HA1-525 was detected by ELISA.Then the HA1-525 was precipitated by PEG20000.After immunization with the HA1-525 , the anti-HA7 antibody in mouse serum was detected by ELISA and hemagglutinin inhibition ( HI) test.Results The HA1-525 was expressed in P.pastoris after being induced with methanol. Western blotting confirmed that there were specific dispersion bands and the molecular weight of HA1-525 decreased to 58 × 103 after being digested by endo H.Anti-HA7 antibody was found in serum of HA1-525 immunized mice and the hemaggluti-nation-inhibition titers reached 1∶700 after the third doses.Conclusion This study shows the HA1-525 expressed in P.pastoris can induce the neutralizing antibody in mice.

OBJECTIVE To investigate the antimicrobial susceptibility of Streptococcus pneumoniae isolated in Yantai and their mechanisms of resistance to macrolides.METHODS Antimicrobial susceptibility of S.pneumoniae was determined by agar dilution method.Phenotypes of macrolide-resistant S.pneumoniae were determined using double disk test with erythromycin and clindamycin disks.ermB And mefE genes were amplified by PCR.RESULTS Among 42 strains of S.pneumoniae,65.0% were intermediate to and no strain was resistant to penicillin.The resistance rates to erythromycin and clindamycin were 93.0%,respectively.Of 41 erythromycin resistantstrains,93.0% were constitutive resistant.ermB Was detected in 40 strains and mefE in 1 strain,both ermB and mefE genes were found in 9 strains.CONCLUSIONS The resistance rate of S.pneumoniae to penicillin is high in Yantai area,the resistance rates to erythromycin and clindamycin are very high.Target modification by ermB methylase is the predominant mechanism in macrolide-resistant S.pneumoniae in Yantai.

Objective:Immunoregulation study of umbilical mesenchymal stem cell (UCMSCs) on allogeneic umbilical cord blood(UCB) CD4+T lymphocytes,which proliferation,apoptosis and the differentiation to CD4+CD25+ regulatory T cell (Treg) in vitro. Methods:Establishing on direct contact or transwell co-culture system,adopt in different proportion of UCMCs with phytohaemag-glutinin (PHA)-activated UCB CD4+T lymphocytes were co-cultured. The proliferation of lymphocyte,percent of CD4+CD25+/CD4+and Foxp3 expression, regulatory T cell marker gene were measured. Apoptosis of CD4+T lymphocytes were observed in the direct contact or transwell coculture system of UCMSCs with desamethason( DXM)-stimulated UCB CD4+T lymphocytes. Results: The UCB CD4+T lymphocytes cocultured with UCMSCs with PHA-activating for 3 days,compared with the UCMSCs free control group,the amount of cells was reduced noticeably(P<0. 05) and the percent of CD4+CD25+in CD4+T lymphocytes and Foxp3 expression significantly in-creased(P<0. 01) in a dose dependent way(P<0. 05). The UCB CD4+T lymphocytes cocultured with UCMSCs with DXM-inducing for 7 days,the apoptosis rate was significantly lower than that of the control group without UCMSCs (P<0. 01). These effects were partially attenuated in transwell coculture but could not be eliminated. Conclusion: UCMSCs are negative effect on UCB CD4+T lymphocytes-mediated immunity effects,and mainly manifested in the regulation on cell proliferate ability and differentiation rather than promoting apoptosis.