Objective Clearing HIV provirus is the key to cure AIDS .The study was to construct the Tre enzyme eukaryotic expression vector and identify its function in specific recognition of loxLTR sequence in HIV provirus . Methods Tre gene was in-serted into eukaryotic expression vector pcDNA 3.1 gene recombination manipulation by genetic recombination techniques including gene synthesis , PCR, restriction enzyme digestion and ligation .EGFPpA-LoxLTR sequence was inserted into pmCherry-N1 vector and was tested by restriction enzyme digestion , PCR and sequencing .Constructed vectors were electroporated into HeLa cells , then using fluorescence microscopy to observe fluorescence intensity changes . Results PCR, restriction enzyme digestion , electrophoresis and sequencing confirmed that Tre enzyme eukaryotic expression vector had been constructed successfully , and it could specifically recog-nize and cut loxLTR sequence after being transfected into Hela cells . Conclusion Constructed Tre enzyme eukaryotic expression vector can be expressed in Hela cells and specifically recognize loxLTR sequence , which has prepared the experimental ground for fur-ther studies of clearing HIV provirus .

OBJECTIVE To investigate the distribution of clinical bacterial isolates and the change in antibiotic resistance spectrum in our hospital from 2005 to 2007.METHODS Data of bacterial susceptibility testing of clinical isolates from the Second Affiliated Hospital in of University of South China from 2005 to 2007 were collected and analyzed by software WHONET25.Results were assessed according to the National Committee for Clinical Laboratory Standards(NCCLS) of America issued in 2005.RESULTS The amount of Gram-negative bacteria decreased and of Gram-positive bacteria increased during this period.The proportion of coagulase negative Staphylococcus(CNS) had been increasing and reached 21.7% in 2007.The proportions of Staphylococcus aureus decreased from 17.6% in 2005 to 13.0% in 2007.Escherichia were the top two bacteria in 2007.The drug resistance rate of staphylococci against penicillin and erythromycin was more than 92.2% and 52.2%,respectively.The oxacillin resistance rate of CNS was 74.5%,significantly higher than that of S.aureus(16.5%).Drug resistance rate of Enterococcus to vancomycin was 1.1%.Gram-negative bacteria were found resistant to meropenem and imipenem.The resistance rate to ampicillin of Klebsiella and Escherichia was very high.CONCLUSIONS The variation of drug resistance and distribution of clinical bacterial isolates in our hospital are related to the improper use of antibiotics.It is very important to select antibiotics correctly according to the results of antibiotics susceptibility tests.

Objective To investigate the prevalence of A2058G or A2059G mutation within 23S rRNA in Treponema pallidum (Tp) from primary syphilis patients chancre samples. Methods Simple PCR was used to screen the positive samples containing Tp DNA. Nested PCR was adopted to amplify the region of the Tp 23S rRNA and the purified amplicons were digested by restriction endonuclease MboⅡand Bsa I respectively and sequenced. Results 39 qualified samples were obtained from 43 chancre samples and all of them were found harboring the A2058G mutation, whereas the A2059G was not detected. Conclusion High frequency of the A2058G mutation within 23S rRNA implicated in macrolide resistance emerges in the circulating Tp in Hengyang. Therefore, macro-lide antibiotics such as azithromycin should be cautiously used as an optional therapy for syphilis.

Objective To investigate the feasibility of C.trachomatis culture in HaCaT human keratinocytes.Methods According to the procedure for C.trachomatis culture in McCoy cells,clinical swab specimens and standard strains of C.trachomatis serotype E were inoculated into HaCaT cells.Iodine staining,a fluorescent monoclonal antibody test and PCR amplification of the endogenous plasmid of C.trachomatis were performed to detect the growth of C.trachomatis in HaCaT cells.Five passages of subculture were carried out for the standard strain of C.trachomatis serotype E in HaCaT cells,and inclusion bodies were counted after each passage.One-factor analysis of variance was conducted by using the software SPSS17 to determine if C.trachomatis was propagated in HaCaT cells.Results Iodine staining showed typical inclusion bodies of C.trachomatis in the cytoplasm of HaCaT cells.Yellow fluorescence-labeled granules were observed in the HaCaT cells under a microscope.Endogenous plasmids of C.rachomatis were successfully amplified by PCR from the infected HaCaT cells.The number of inclusions in HaCaT cells gradually increased at passage 1 through 5.Conclusions C.trachomatis is successfully cultivated in HaCaT cells in vitro,and the standard strain of C.trachomatis serotype E can propagate in HaCaT cells.

Objective To investigate the binding protein of chlamydiaphage phiCPG1 capsid protein Vp1 on chlamydia trachomatis outer membrane.Methods The bacterium with recombinant plasmid Vp1/pet30a( + ) was induced.The expressed protein was purified by gel recycling.FarWestern blot was utilized to' investigate the binding protein of Vp1 on chlamydial outer membrane,including recombinant polymorphic outer membrane protein (rPmp) and major outer membrane protein (MOMP).Results The recombinant protein Vp1 was successfully expressed in E.coli.Monoclonal antibody against Vp1 was used as primary antibody in Western blot,and no specific band was present,which indicated that the monoclonal antibody did not specifically bind with any rPmp.Far-Western blot results showed that there was an obvious band for the rPmpI,but no specific band for other rPmp and MOMP,which suggested that Vp1 could specifically bind with rPmpI protein on the chlamydial outer membrane of serotype D.Conclusions There is a binding site of Vp1 on the chlamydia trachomatis outer membrane.Vp1 may play an important role in the interaction between the chlamydiaphage and the chlamydiae.

Objective:This thesis aims to construct the evaluation index system ethics management in Beijing medical institutions for application research.Method:The author applied the evaluation system which was agreed and adjusted by expert groups in four medical and health institutions.Result:After three rounds of expert groups have reached the agreement,the Evaluation Index System on Human Research Ethics Management in Beijing Medi-cal Institutions is established,which contains 6 first-class indicators,16 second-class indicators and 39 third-class indicators.The application on four medical and health institutions show that the evaluation index system is practical,operable and is of high degree of distinction.Conclusion:The evaluation index system is scientific,rea-sonable,practical and is of high degree of validity,credibility and distinction.

OBJECTIVETo compare the efficacy of telbivudine monotherapy and telbivudine combination therapy with adefovir in patients with nucleos(t)ide-naive chronic hepatitis B, high-level hepatitis B virus (HBV) load and hepatitis B e antigen (HBeAg)-positivity, and to explore the relationship between treatment regimen adherence and treatment outcomes.

METODSA retrospective study was performed with 123 HBeAg-positive, high-level viral load (HBV DNA≥10(7) copies/ml), nucleos(t)ide-naive chronic hepatitis B patients. Fifty-three of the patients received combination therapy with telbivudine and adefovir dipivoxil,while 70 patients received the telbivudine monotherapy. All patients were tested for rates of conversion to HBV DNA-negative status,alanine aminotransferase (ALT) normalization, HBeAg seroconversion, drug resistance, and side effects at treatment weeks 12, 24, and 48. Treatment regimen adherence was assessed through self-reporting, and interviews were used to explore the relationships to treatment outcomes. The chisquare test, t test and Fisher's exact test were used for statistical analyses.

RESULTSThe rates of HBV DNA negative conversion in the combination group at treatment weeks 12, 24 and 48 were 62.3% (33/53), 88.7% (47/53) and 94.3% (50/53) and were significantly different from those in the monotherapy group at weeks 12 and 24.The rates of ALT normalization were significantly different between the two groups at treatment week 12 (94.3% vs. 77.1%). The rate of HBeAg seroconversion in the combination group at treatment week 48 was 39.6%, and significantly different than that of the monotherapy group. The rates of drug-resistance in the combination and monotherapy groups at treatment week 48 were 3.8% and 11.4%, and the proportion of non-adherence to the treatment regimen was 53.3%, which significantly affected treatment outcome. No side effects occurred in either treatment group.

CONCLUSIONTelbivudine combination treatment with adefovir was more effective than telbivudine monotherapy and elicited a low drug resistance rate in nucleos(t)idenaive chronic hepatitis B patients with high-level HBV load and HBeAg-positivity. Adherence to the therapy regimen was a key factor influencing treatment outcomes.

Adenine ; analogs & derivatives ; Alanine Transaminase ; Drug Therapy, Combination ; Hepatitis B e Antigens ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Organophosphonates ; Retrospective Studies ; Thymidine ; analogs & derivatives ; Treatment Outcome ; Viral Load

STUDY DESIGN: A prospective imaging study. PURPOSE: To characterize the distribution of the global sagittal postural patterns in asymptomatic Chinese adults using Roussouly classification. OVERVIEW OF LITERATURE: The norms of sagittal parameters in asymptomatic Chinese population have been previously described, but no report described their global sagittal postural patterns as characterized by Roussouly classification. METHODS: A cohort of 272 asymptomatic Chinese adults was recruited. Data was assimilated by reviewing the films for each subject. Sagittal parameters were measured and sagittal postural patterns were then determined according to Roussouly classification. The pattern distributions were compared across genders within the study cohort. We also compared the data across different ethnicities from our study and a previous study to further characterize Chinese sagittal postures. RESULTS: The cohort included 161 males and 111 females, with mean age of 23.2±4.4 years. The average descriptive results were as below: pelvic incidence (PI) 46.4°±9.6°, thoracic kyphosis (TK) 24.2°±9.0°, lumbar lordosis (LL) 50.6°±10.6°, sacral slope (SS) 37.2°±7.6°, pelvic tilt (PT) 9.4°±6.8°, spinosacral angle (SSA) 131.1°±7.5° and sagittal vertical axis (SVA) 17.24±32.36 mm. Despite a significant difference between two genders in LL, PI, SSA, and SVA, no difference was found in the distribution of Roussouly types among them. 47.8% of our cohort belonged to Roussouly type 3, while type 1, 2 and 4 comprised 23.2%, 14.0% and 15.1% of the subjects, respectively. Roussouly classification was capable of categorizing sagittal parameters except for the PT. This study also found that 4.4% of the recruited subjects belonged to the C7-anterior subgroup. CONCLUSIONS: From a characterization of the sagittal postural patterns of asymptomatic Chinese adults using Roussouly classification, the distribution was similar between Chinese males and females; however, from a cross-study comparison, it was different between asymptomatic Chinese and Caucasian adults, with a higher proportion of Roussouly type 3 in Chinese adults.
Adult* ; Animals ; Asian Continental Ancestry Group* ; Axis, Cervical Vertebra ; Classification ; Cohort Studies ; Female ; Humans ; Incidence ; Kyphosis ; Lordosis ; Male ; Posture ; Prospective Studies

Objective To investigate the distribution and changes of pathogens in intensive care unit of our hospital from 2010 to 2015,and provide treatment suggestions for infection.Methods The data of pathogens and multi-drug resistant organisms(MDROs)in ICU from 2010 to 2015 were analyzed retrospectively by WHONET5.6,Microsoft Office Excel2007 and SPSS19.0.Results The numbers of pathogens in ICU were increasing year by year while the top six were Acinetobacter baumanni,Pseudomonas aeruginosa,Klebsiella pneumoniae,Escherichia coli,Staphylococcus aureus and Candida albicans.The dominant MDROs were multi-drug resistant Acinetobacter baumanni,multi-drug resistant Pseudomonas aeruginosa,extended spectrum β lactamase producing Escherichia coli and Klebsiella pneumonia,methicillin-resistant coagulase negative Staph-ylococcus and methicillin-resistant Staphylococcus aureus,etc.There were statistically dramatic significant differences of most MDROs during the six years(P<0.01).Conclusion The pathogens of ICU were mainly bacteria related to the healthcare associated infection,and the multi-drug resistance was obviously in dynamic change as well as most pathogens.As a suggestion,antibiotic agents should be used rationally according to the antimicrobial susceptibility results for treating the MDROs.

Through the questionnaire survey on the current situation of human research ethical management in Beijing medical and health institutions, this paper analyzed the problems existing in ethical management of Beijing medical and health institutions and put forward corresponding countermeasures and suggestions. The results showed that overall status of human research ethical management in Beijing medical and health institutions was satisfying. But the ethical management levels of different types of medical and health institutions were quite different, and medical and health institutions, health family planning administration departments and academic teams should all make corresponding contributions.