Purpose:To clone and construct an eukaryotic expressive vector of ribonuclease inhibitor (RI) gene ,as well as to observe the effects of the transfected pLNCX-ri on the growth of C6 glioma cells.Methods:A segment of RI gene of 1.4 kb was obtained by Nde I/Xho digestion and cloned into pLNCX. Transfective agent and selective antibiotic were lipofect AMINE and G418 respectively. The expression of pLNCX-ri in C6 glioma cells was detected by Western blotting. And SD rats were inoculated by the transfected C6 glioma cells.Results:An eukaryotic expressive vector of RI gene was constructed successfully. RI content was remarkably higher in the transfected cells than that of in the untransfected cells. After SD rats were inoculated by the transfected C6 glioma cells,the tumorigenic time was prolonged, the tumor weight was reduced and the density of tumor vessels was notably decreased. Conclusions:These results indicated that RI gene powerfully inhibited the growth of C6 glioma cells via decreasing tumor vessels formation.

Objective To evaluate the effect of medical hygiene wipes on disinfecting object surface and compliance to use in an intensive care unit (ICU).Methods 30 beds in the ICU at a hospital were randomly selected and divided into three groups:A,B and C,each group had 10 beds.Group A were only provided hygiene wipes,group B used towel soaked in chlorine-containing disinfectant (disinfected towel),group C were provided both medical hygiene wipes and disinfected towel for selected use.Compliance to different disinfection methods,and disinfectant efficacy on object surface were compared.Results The rate of compliance to disinfection in group A was higher than group B (78.78% vs 42.12%,χ2 =68.56,P <0.05);except medical disposal trolley and ventilator,compliance to disinfection of the other object surface in group A were all higher than group B (all P <0.05).Except ventilator, adenosis triphosphate (ATP)values before and after disinfection of object surface in group A and B were both significantly different (both P <0.05 ),disinfectant efficacy of group A was superior to group B.Group C used medical hygiene wipes more than disinfected towel ([836 times,91 .57%]vs [77 times,8.43%]).Nurses spent (20.15+ 3.00 )minutes on preparing disinfectant every day,and (15.02 ± 2.00 )minutes on washing used disinfected towel,while hygiene wipes could be used immediately,discarded after use,and was timesaving. Conclusion Use of medical hygiene wipes is convenient,rapid,and without irritating odor,it also improves disinfection effect on object surface and ICU health care workers’compliance to disinfection.

Objective To observe the growth situation of Blastocystis hominis in vitro and select the optimal method for culti?vation of B. hominis in different media. Methods Ten positive stools with B. hominis were inoculated in three different media for cultivating,namely 1640,Jone’s medium and vitro medium. And the stools with good growth status and high quantities of B. hominis were chosen to inoculate in the three media with equal amount after subcultivation,and the number of B. hominis was counted every 24 h for ten days,and the morphological changes and growth status were also observed. Results The densities of B. hominis in the 1640 and Jone’s medium were higher than that in the vitro medium 48 h after the inoculation. The same stool sample was inoculated to the three different media and observed for ten days,and the results indicated that the growth of B. homi?nis presented regular changes in the three media,the growth peaks were on the third,sixth and ninth day post inoculation;and the density of B. hominis was the highest in the Jone’s medium. The morphology of B. hominis was the clearest and most dynamic in the vitro medium,while various reproductive forms were observed in the Jone’s medium. Conclusion Jone’s medium is suitable for the growth of B. hominis and can be the first choice for the cultivation of B. hominis in vitro,and vitro medium is the best medium for observing the growth situation of B. hominis.

Objective To evaluate the roles of Toll-like receptor 2 (TLR2),TLR4 and myeloid differentiation factor 88 (MyD88) in immune recognition and immune mediation in sporotrichosis by detecting their expressions in skin lesions of sporotrichosis.Methods Biopsy specimens were obtained from the skin lesions of 19 patients with sporotrichosis and normal skin of 12 healthy human controls.Immunohistochemistry was performed to observe the expressions of TLR4 and MyD88,and real-time fluorescence-based quantitative PCR to quantify the expressions of TLR2 and MyD88 mRNAs.Data were expressed as mean ± standard error.Statistical analysis was done with the SPSS17.0 software.Independent samples t-test was conducted to compare the parameters between the lesional and control specimens.A p-value of less than 0.05 was considered to be statistically significant.Results TLR4 and MyD88 were mainly observed in the whole epidermis except the stratum corneum as well as plasma cells and lymphocytes in the dermis and subcutaneous tissue in lesional skin.However,TLR4 was nearly absent and MyD88 was weakly expressed in the dermis and subcutaneous tissue in normal skin.The expression levels of TLR4 and MyD88 were significantly higher in the lesional skin than in the control skin (TLR4,63.767 ± 3.829 vs.5.167 ± 3.246,t =4.82,P ＜ 0.05; MyD88,57.236 ± 4.744 vs.10.588 ± 1.640,t =3.30,P ＜ 0.05).Similarly,the lesional skin showed significantly stronger expressions of TLR2 and MyD88 mRNAs compared with the normal skin (TLR2,1.974 ± 1.452 vs.1.430 ± 1.073,P ＜ 0.05; MyD88,2.028 ± 2.061 vs.0.688 ± 0.422,P ＜ 0.05).Conclusion Sporothrix may induce the development of sporotrichosis by interacting with host immune system via TLR signaling pathways.

Objective To investigate the effect of atorvastatin on expression of ABCA1(mRNA,protein) and LXR? mRNA in THP-1 macrophages induced by PMA.Methods Cultured THP-1 cells were induced to differentiate into macrophages by PMA for 48 hours.The macrophages were incubated with atorvastatin in different concentions for 24 or 48 hours.We determined the changes of ABCA1 mRNA,protein and LXR? mRNA by reverse trancriptase polymerase chain reaction(RT-PCR) and immuno-histochemistry.Results The expression level of ABCA1 mRNA(ratio of relative expression 1.21 vs 1.48) and protein as well as LXR? mRNA(0.87 vs1.12) were decreased in THP-1macrophages when cultured with atorvastatin(10 ?mol/L) for 24 h.Conclusion Atorvastatin inhibits the expression of ABCA1 mRNA and protein as well as LXR? mRNA of THP-1macrophages in vitro.

Objective:To investigate associations between glycosylated hemoglobin (HbA1c) levels and nonalcoholic fatty liver disease (NAFLD) in population who underwent periodic heath examination.Methods:A total of 912 subjects were selected from the health examination adults in University of Hong Kong Shenzhen Hospital from June to November 2013,and they were assigned into a NAFLD group,an else hepatic disease group,and a control group.The physical examination results of these subjects were analyzed retrospectively.Results:The prevalence of NAFLD was 29.7％.The related factors such as age,body mass index,and the levels of systolic blood pressure,diastolic blood pressure,total cholesterol,triglyceride,low density lipoprotein cholesterol,fasting glucose,HbA1c and alanine aminotransferase in the NAFLD group were higher than those in the else hepatic disease group and the control group (P＜0.05).Aspartate aminotransferase levels in the NAFLD group were higher than those in the control group (P＜0.05).High density lipoprotein cholesterol levels in the NAFLD group were lower than those in the else hepatic disease group and the control group (P＜0.05).According to the quartile of HbAlc level,these subjects were divided into 4 groups,Q1,Q2,Q3,and Q4,and the prevalence of NAFLD in the Q1,Q2,Q3,and Q4 were 17.5％,21.5％,31％,and 59.4％,respectively.The prevalence of NAFLD increased along with the increase in the level of HbAlc (x2=100.092,P＜0.05).Logistic regression analysis showed that the risk factor for NAFLD was increased in the subjects of Q4 as compared with the subjects of Q1 after adjusting for the related factors including age,gender,body mass index,etc.Conclusion:There is a positive correlation between the prevalence of NAFLD and HbAlc level,and the risk of NAFLD increases with the elevating level of HbA1c.High HbA1c level is an independent risk factor for NAFLD.Thus it should be listed as a routine medical test in the health examination.

Objective: To construct the public health performance evaluation index system for the county medical community, so as to provide reference for the assessment of the public health work in the county medical community. Methods: According to the 2019 Public Health Tasks of Zhejiang Province and the current status of the county medical community's public health work, a framework was developed. Twenty experts from universities, CDCs, and county medical community were invited to construct the index system after two rounds of Delphi expert consultation, and the index weight was determined by the analytic hierarchy process. Results : The experts aged ( 46.10±6.41 ) years and worked for ( 23.85±7.37 ) years, all of whom had a bachelor degree or above, and 18 had associate senior titles or above. The response rates of two rounds of consultation was both 100.00%; the authority coefficient was 0.811; the values of Kendall's W all had statistical significance ( P<0.05 ) , which in the second round were higher than those in the first round. The public health performance evaluation index system for county medical community finally included 10 first-level indexes, among which priority diseases surveillance and management weighed most ( 0.165 7 ) ; 32 second-level indexes, among which food-borne diseases surveillance, infectious diseases/public health emergencies reporting and infectious diseases/public health emergencies response weighed the top three ( 0.071 5, 0.064 7, 0.063 8 ); 120 third-level indexes, among which the timely reporting rate of food safety incidents, the reporting rate of infectious diseases and the information response rate of infectious diseases/public health emergencies weighed the top three ( 0.039 4, 0.022 1, 0.021 7 ). Conclusion The public health performance evaluation index system of the county medical community has been established, which can provide reference for the health administrative departments.

Objective To explore the effects of Living with Hope Program(LWHP) on family caregivers of patients with advanced lung cancer.Methods Totally 60 family caregivers of patients with advanced lung cancer were randomly divided into the experimental group and the control group.The experimental group received LWHP intervention and routine care knowledge education,the control group only received routine care knowledge education.Hope,self-efficacy,anxiety and depression,and quality of life were evaluated at the first week,the second week,the first month,and the third month.Differences between two groups were compared using variance analysis of repeated measurements.Results The self-evaluation of the intervention was 76.6％,and the main effects of hope level,selfefficacy,anxiety and depression,and mental health were statistically significant (P＜0.05);the time effect on hope level and self-efficacy were statistically significant (P＜0.05);there were interaction effects between intervention and time on hope level,self-efficacy,anxiety and depression,and mental health (P＜0.05).Conclusion LWHP can effectively improve hope level,self-efficacy and mental health status,and alleviate anxiety and depression of family caregivers of patients with advanced lung cancer.

Objective To evaluate the clinical value of ultrasonography in the diagnosis of hermaphroditism.Methods Ultrasonographic appearances of 45 cases with hermaphroditism were reviewed retrospectively,and the results were compared with clinical data.Results All cases were confirmed pathologically.5 cases showed true hermaphroditism (11.1％),14 cases showed male pseudohermaphroditism (31.1 ％),24 cases showed female pseudohermaphroditism (53.3 ％),2 cases showed gonadal dysgenesis(4.4％).Compared with pathological results in the ultrasound examination,4 cases showed true hermaphroditism,the coincidence rate was 80％.11 cases showed male pseudohermaphroditism,the coincidence rate was 78.6％.20 cases showed female pseudohermaphroditism,the coincidence rate was 83.3％.1 cases showed gonadal dysgenesis,the coincidence rate was 50.0％.Conclusions Ultrasonography can not only evaluate internal reproductive organs in the patients,but also estimate the site,size,morphology and structure,which provides important imaging evidence for the clinical diagnosis and treatment.

OBJECTIVETo analyze the influence of optimal codon usage on the expression levels and immunogenicity of DNA vaccines, encoding the human papillomavirus type 6b (HPV 6b) E7 gene.

METHODSThe full length E7 gene of HPV 6b was modified to substitute human preferred codon for rarely used codon, and three mutations were introduced into the pRB binding site of HPV 6b E7 to eliminate its transformation potential. The codon optimized and mutated E7 gene (hu-mE7) were cloned into the Kpn I and EcoR I site of the pcDNA3 mammalian expression vector, the in vitro expression of the hu-mE7 gene and the immunogenicity of hu-mE7 DNA vaccine were compared with the wt-E7gene.

RESULTSThe in vitro expression of pcDNA3-hu-mE7 was much higher than the classical wt-E7 plasmid in monkey COS-1 cell line. Mice immunized intramuscularly with the pcDNA3-hu-mE7 showed that the codon modified E7 gene induced a stronger IFN-gamma ratios than the wt-E7 gene.

CONCLUSIONSThese results suggest that the optimized codon usage contributes to the enhancement of gene expression and immunogenicity of HPV 6b E7 gene.

Animals ; Cell Line ; Codon ; genetics ; Female ; Gene Expression ; Genes, Viral ; genetics ; Genetic Vectors ; Mice ; Papillomaviridae ; genetics ; Papillomavirus Vaccines ; Transfection ; Vaccines, DNA ; immunology ; Viral Proteins ; biosynthesis ; genetics ; Viral Vaccines ; immunology