We wished to screen the cell line that stably expresses the HPV18E5 protein, and to ascertain the influence of HPV18E5 protein on cell proliferation and the cell cycle. The HPV18E5 gene was amplified by the polymerase chain reaction. Then, the His-tag pSecTag-HPV18E5 eukaryotic expression vector was constructed by digestion ligation and connection. The recombinant plasmid was transfected into Balb/c3T3 cells with lipofectamine, and positive cell lines were screened by a culture medium containing bleomycin. HPV18E5 expression in cells was confirmed by western blotting and immuno-enzymatic methods. The influence of HPV18E5 on cell proliferation and the cell cycle were detected by Cell Counting Kit-8 and flow cytometry, respectively. The pSecTag-HPV18E5 eukaryotic expression vector was constructed. After 21-day selection in a culture medium containing 400 μg/mL bleomycin, stably expressing HPV18E5 protein cells were harvested. Compared with control groups, cell proliferation in HPV18E5 stably expressed cells was obviously increased, as was the S phase in the cell cycle. Our results suggested that HPV18E5 influences cell proliferation and the cell cycle. Our study has laid the foundation of the biologic properties of HPV18E5 protein, which will aid further studies on the mechanism of action of carcinogenesis.
Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Human papillomavirus 18 ; genetics ; metabolism ; Humans ; Oncogene Proteins, Viral ; genetics ; metabolism ; Papillomavirus Infections ; physiopathology ; virology ; Transfection

Objective To investigate the effects of polyi:c on angiogenesis in mouse prostate carcinoma tissue and their possible mechanisms.Methods Mouse prostate carcinoma models were randomly divided into two groups according to tumor weight:control group and polyi:c group.After treatment for 7 times,the mice were sacrificed and the tumor tissues were cut for weighing,calculating the tumor inhibitory rate and tumor index.Hematoxylin-eosin staining and immunohistochemical staining were performed to observate the morphological changes of prostate carcinoma tissues,distribution of vasa and expressions of vascular endothelial growth factor (VEGF) and endothelial nitric-oxide synthase (eNOS).Results In polyi:c group,the mean tumor inhibitory rate was 67.85% and the tumor index was(5.42?0.17)%;in control group,the tumor index was(14.45?1.06)%; there was significant differences between polyi:c group and control group( P

Objective To explore the role of state-level continuing education courses of oper-ation-room nursing in improving the nursing safety cognition in nursing staff. Methods The state-level continuing education programs about the operating room safety and quality management were held for five sessions from Jan 2007 to Dec 2012, in which a total of 641 people participated. All participants were examined before and after each training session. The scores of the examination were utilized to evaluate the cognitive level of the operation-room nusring safety. Results Compared with before training, the students' scores after training were significantly elevated(P<0.05); compared with that after one training session, the students' scores after four and five training sessions were markedly increased(P<0.05). After hospitals sending representatives to participate in the training, their operat-ing room adverse events were decreasing. Conclusion Holding the state-level continuing education programs about operating room safety and quality management helps the nurse staff improve the cogni-tive level of operation-room nursing safety, and their cognitive level can be reinforced after receiving repeated training sessions. Moreover, holding the continuing education programs can decrease the in-cidence of adverse events and the complications in the operation-room.

OBJECTIVE: To explore whether pre-administration of Ganoderma lucidum spore (GASP) can reduce incidence of neural tube defects (NTDs) induced by all-trans retinoic acid (ATRA) in pregnant mice. METHODS: Twenty pregnant mice were randomly divided into four groups: normal control group, solvent-treated group, ATRA-induced group, and GASP-treated plus ATRA-induced group. GASP solution, which was prepared with solvent (sodium carboxymethyl cellulose), was fed to the pregnant mice in the GASP-treated plus ATRA-induced group twice a day from embryo (E) 0 d to E10.5 d. The same dose of solvent was given to the pregnant mice in the solvent-treated group. At E7.75 d, ATRA (50 mg/kg) was given to the pregnant mice in both ATRA-induced group and GASP-treated plus ATRA-induced group for single time. Embryos were sampled from pregnant mice at E10.5 d. Then the incidence rate of NTDs in mouse embryo was calculated and the crown-rump length of mouse embryo was measured. The positive rate of nestin expression and the distribution of cell cycle of embryonic neural tube neuroepithelial cells were detected by histochemical staining technique and flow cytometry respectively. Reverse transcription-polymerase chain reaction method was used to detect the gene expressions of cyclin-dependent protein kinase 2 (Cdk2) and Cdk4 mRNAs. RESULTS: The incidence rates of NTDs in mouse embryos in the ATRA-induced group and the GASP-treated plus ATRA-induced group were 79.41% and 21.67% respectively, while the crown-rump length of mouse embryos in these two groups were (3.62+/-1.27) mm and (5.84+/-0.92) mm respectively. The positive rate of nestin expression in embryonic neural tube neuroepithelial cells of mouse embryo at E10.5 d in the ATRA-induced group was 32.44%, while that in the GASP-treated plus ATRA-induced group was 77.65%. The cell cycle of embryonic neural tube neuroepithelial cells was obviously arrested at G(0)/G(1) phase in the ATRA-induced group as compared with that in the GASP-treated plus ATRA-induced group. The Cdk4 mRNA was transcripted at a high level in embryonic neural tube in the GASP-treated plus ATRA-induced group, but the Cdk2 mRNA was not detected in this group. CONCLUSION: Pre-administration of GASP can reduce the incidence of NTDs induced by ATRA in pregnant mice.

Objective To explore changes of the immune function of red blood cells in gastric disease.Methods RBC C3b receptor rosette(RBCC3bRR)and RBC immune complex rosette(RBCICR)were tested in chronic gastritis group(n =103),duodenal ball ulcer group(n =75)and control group(n =30)to evaluate the immune function of RBC.Results RBCC3bRR were(20.83 ± 5.16)％ in the control group,(16.26 ±5.17)％ in the chronic gastritis group and(13.65 ± 5.19)％ in the duodenal ball ulcer group.RBCICR were respectively(7.63 ± 4.09)％,(10.59 + 4.45)％ and(10.04 ± 4.13)％ in these three groups.RBCC3bRR of chronic gastritis and duodenal ball ulcer were lower than control group(P ＜0.0l),while RBCICR were higher than normal control group(P ＜0.05 and P ＜0.01 respectively).There was no significant difference on RBCC3bRR and RBCICR between HP negative chronic gastritis and HP negative duodenal ball ulcer and between HP positive chronic gastritis and HP positive duodenal ball ulcer(P ＞ 0.05).RBCC3bRR of HP positive chronic gastritis and duodenal ball ulcer was significantly lower than that of HP negative(P ＜ 0.05and P ＜ 0.01 respectively),RBCICR significantly higher than that of HP negative(P ＜ 0.01).After HP eradication,RBCC3bRR of patients with chronic gastritis and duodenal ball ulcer was increased significantly (P＜0.05 and P ＜ 0.01 respectively).RBCICR was significantly lower than before treatment(P ＜ 0.01).Conclusion HP infection,chronic gastritis and duodenal ulcer can decrease the immune function of red blood cells.

OBJECTIVE:To evaluation of the efficacy and safety of azithromycin in the treatment of acute infections METHODS:A randomized controlled multicenter clinical study was used to compare the clinical efficacy of azithromycin(200mg q d for 3～5 days) with erythromycin(500mg b i d for 5 days) RESULTS:The cure rate,effective rate,and bacterial clearance rate were 76 7%,95 0%,and 94 3% respectively for azithromycin group and were 48 3%,76 7%,and 77 4% respectively for erythromycin group with a side-effect incidence of 10 0% for azithromycin group and 39 3% for erythromycin group CONCLUSION:Azithromycin is superior to erythromycin in clinical efficacy,safety and side-effect incidence

Objective To explore the relationship between serum 25-hydroxy vitamin D [25(OH)D] concentrations and the risk of Henoch-Schonlein purpura (HSP).Methods A case control study was designed.Serum 25 (OH)D concentrations were measured by radioimmunoassay in 214 participants,including 53 H SP patients and 161 status-matched healthy controls.Information concerning demographic data,genetic,background,and environmental exposures was collected using questionnaire.The study participants were divided into four groups according to quartile range of 25(OH)D concentration and logistic regression modeling was used to evaluate the relation with HSP risk by estimating odds ratios(OR)and 95％confidence intervals(CI).Results The HSP group had a significantly lower concentration of 25(OH)D than the control group (the median in the HSP group was 11.4 ng/mL;controls:15.36 ng/mL,P＜0.05).When the first interval was set as the reference level,the OR (95 ％ CI) of the second,third,and fourth intervals were:0.468(0.341-0.771),0.442(0.302-0.627),0.339 (0.199-0.501).After adjusting the analysis for the presence of pathogenic related confounding fact OR,the OR(95％CI)of the second,third,and fourth intervals were:0.459(0.333-0.741),0.408(0.317-0.611),0.387 (0.221-0.517).The 25 (OH) D level was inversely correlated with the risk of HSP(P＜ 0.05).Conclusion The risk of HSP was decreased with the increase of serum 25 (OH) D concentration,25 (OH) D may be a protection factor in the pathogenesis of HSP.

Improving medical students' calculation ability in statistics has become the focus and difficulty of medical statistics course teaching, and its application relies heavily on statistical calculation software. In order to explore a new teaching approach which combined the advantages of traditional method and web-based calculation, we intended to build a web computing platform applying the Browser/Server (B/S) mode based on the campus network, and to revise the current syllabus of medical statistics, as well as to create a virtual web-lab containing a typical case library. Practice has proved that the new mode effec-tively improved the practice capability of students and changed the traditional teacher-centered teaching approach.

OBJECTIVE To analyze the clinical features of 8 patients with sigmoid sinus diverticulum and to improve the level of diagnosis and treatment. METHODS The tinnitus patients were asked in detail about the history, and performed a series of routine hearing tests (pure tone audiometry, acoustic impedance audiometry, otoacoustic emission inspection and brainstem auditory evoked potential (BAEP) etc.) and temporal bone HRCT scan. Eight cases were diagnosed with sigmoid sinus diverticulum, in which six cases undertook a surgery to reconstruct the sinus wall while the other 2 cases chose conservative treatment. RESULTS The diagnosis of 6 cases was proved to be correct by surgery, and the pulsatile tinnitus disappeared after reconstructing the sinus wall. No one recurred after following up from 2 to 12 months. Two cases of conservative treatment had no obvious improvement in symptoms. CONCLUSION Clinical manifestation, hearing examination, temporal bone CT/HRCT and Cranial CTA/CTV contributed to the diagnosis of sigmoid sinus diverticulum. Sinus wall reconstruction is a better approach.

Objective To establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on hemolysin gene(vvhA)that coding cytolysin.Method By using software Primer Express, the PCR primers and TaqMan probe,which located in the conserved region of vvhA gene sequence,were designed for establishment of a TaqMan real-time fluorescent quantitative PCR to detect 100 bp amplicon from V.vulnificus DNA.A recombinant plasmid pMD19-vvhA100 as a positive control during detection was constructed using gene cloning technique.Minimal amplification cycles(Ct value)and fluorescence intensity enhancement (△Rn value)were used as observing index to optimize the reaction conditions of the TaqMan real-time fluorescent quantitative PCR.The DNAs with different concentrations from V.vulnificus and other eight bacteria and pMD19- vvhA100 were applied as templates to determine the specificity,sensitivity and reappearance of the TaqMan real- time fluorescent quantitative PCR.ICR mice were intraperitoneally,subcutaneously and orally infected with V. vulnificus,respectively.The detection effect of the TaqMan real-time fluorescent quantitative PCR was measured using the specimens of peripheral blood,subcutaneous tissue and intestinal content collected from the infected mice.Results The established TaqMan real-time fluorescent quantitative PCR showed positive results only for V. vulnificus DNA and pMD19-vvhA100.The detection effectiveness of the TaqMan real-time fluorescent quantitative PCR was as high as 0.01 ng of V.vulnificus DNA or 103 copies of pMD19-vvhA100.The SD values of the detection results repeated for three times using pMD19-vvhA100 with different concentrations were lease than 0.79. The detection results of TaqMan real-time fluorescent quantitative PCR were positive for all the specimens of peripheral blood and subcutaneous tissue.Conclusions The TaqMan real-time fluorescent quantitative PCR established in this study for V.vulnificus vvhA gene detection has advantages such as quickness,stability, sensitivity and specificity,indicating this method can be used for clinical laboratory diagnosis of septicemia and wound infection caused by V.vulnificus.