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Objective To construct the prokaryotic expression vector of human esophageal cancer related gene 4 (ECRG4), to purify the recombinant ECRG4 protein and to verify the biological function of the recombinant ECRG4 protein. Methods DNA recombination technology was utilized to construct the ECRG4 protein prokaryotic expression vector. The recombinant ECRG4 protein was purified with the transformation of escherichia coli. Then the purity of the recombinant ECRG4 protein was examined by using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Furthermore, esophageal cancer EC-18 cell line was treated by recombinant ECRG4 protein (10 μg/ml) for phosphate buffer (PBS) 48 h respectively, and the tumor cell cycle change was examined by using flow cytometry. Results SDS-PAGE analysis showed that the high purity of recombinant ECRG4 protein was obtained and the ECRG4 protein prokaryotic expression vector was successfully constructed. The cell proportion of G1 phase in PBS control group was lower than that in the recombinant ECRG4 protein group [(60.4 ±2.1) ％ vs. (71.6 ±1.8) ％; t= 25.695, P= 0.002]. The cell proportion of S phase in PBS control group was higher than that in the recombinant ECRG4 protein group [(24.6±1.4) ％ vs. (16.5±1.0) ％; t= 36.905, P= 0.001]. The cell proportion of G2/M phase in PBS control group was higher than that in the recombinant ECRG4 protein group [(15.0 ±1.1) ％ vs. (11.9 ±0.8) ％; t=6.471, P=0.023], which indicated that the recombinant ECRG4 protein could induce the G1 phase arrest of EC-18 cells. Conclusion The ECRG4 protein prokaryotic expression vector is successfully constructed. And the recombinant ECRG4 protein has an active biological function in esophageal carcinoma.
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Objective To learn customer satisfaction for health management centers at public hospitals in Hangzhou. Methods From July to September 2015, 660 customers from the health management centers of 6 public hospitals in Hangzhou were randomly selected for questionnaire survey. The survey included their satisfaction for the technical attitude, service content design, waiting time, environment facilities, management regulations, and service charges. Univariate analysis was carried out on the satisfaction scores of customer demographics. Two-class logistic regression was used to analyze the factors affecting customer satisfaction. Results The average customer satisfaction score was 3. 62 ± 0. 55. Their satisfaction with technical attitude, service content design, environmental facilities and management regulations was higher. They were less satisfied with waiting time(3. 20 ± 0. 85) and service charge(3. 36 ± 0. 71). Conclusions Overall satisfaction of customers for such centers is high. In the future, we should further strengthen the information management and procedures of these centers, and link health management services with commercial insurance, for less economic burden on the people.
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Objective:To evaluate the clinical effects of short fiber ribbon combined with resin bonding technology for the treatment of food impaction between posterior teeth. Methods:98 cases of vertical food impaction between posterior teeth( total of 135 vertical food impaction units) were included. 73 units were treated by short quartz fiber ribbon combined with resin bonding technology( SQFRB) and 63 by resin bonding(RB). 12, 24 and 36 months after restoration, clinical effects were evaluated referring to the Modified United States Public Health Service (USPHS) Criteria, data were statistically analyzed. Results:12, 24 and 36 months after treatment the cure rate of SQFRB was 97. 3%, 97. 3% and 95. 9%, inefficacy rate was 0, 0 and 0;the cure rate of RB was 85. 5%, 82. 2% and 82. 2%, the inefficacy rate was 4. 8%, 11. 3% and 12. 9%, respectively(between groups, P<0. 05). Conclusion:Minimally inva-sive restorations using short fiber ribbon combined with resin bonding technology is effective in the treatment of vertical food impaction between posterior teeth.
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Aim Toresearchthemolecularmecha-nisms of DADS-induced apoptosis in human leukemia K562cells.Methods Cellviabilitywasmeasuredby MTT. Levels of DADS-induced ROS were measured by 2ˊ, 7ˊ-dichlorofluorescein diacetate ( DCFH-DA) fluo-rescence. DADS-induced mRNA levels of components of the NADPH oxidase were detected by Real-time PCR. The combination of protein Rac2 and p67phox was measured by immunoprecipitation assays. Flow cy-tometry methods were used to determine the percentage of apoptosis cells. DADS-induced Rac2 levels were measuredbyWesternblot.Results TheDADS-trea-ted K562 cells showed a dose-and time-dependent de-crease in cell viability and proliferation. There was sig-nificant up-regulation of the mRNA level of components of the NADPH oxidase complex in K562 cells after treatment with 6 mg·L-1 DADS for 6 h. Western blot results revealed that, compared with the control group, there was a significant up-regulation of Rac2 protein in K562 cells treated with 5. 0 and 10. 0 mg·L-1 DADS for 24h. And Rac2 combined with p67phox in DADS-induced apoptosis in K562 cells. PMA markedly in-creased the percentage of apoptotic cells, and DPI re-duced the percentage of apoptotic cells in DADS-in-duced K562 cells. Levels of DADS-induced ROS, also showed enhancement when exposed in PMA, but there was no DADS-induced ROS production evident when exposed in DPI in DADS induced K562 cells. Conclu-sions TheseresultsindicatethatNADPHoxidaseis the main source of DADS-induced ROS production. Diallyl disulfide induces apoptosis in human leukemia K562 cells through activation of NADPH oxidase.