Objective To establish a method for efficient,accurate genotyping and nucleoside drug-resistant mutation analysis for hepatitis B virus ( HBV ).Methods The 48 HBV serum samples were collected from the Third People's Hospital of Taiyuan from July to August 2011,and HBV DNA were extracted using the commercial kit.The HBV whole genome and P gene were amplified and sequenced.Each HBV sample was genotyped by both constructing phylogenetic trees and genotyping software analysis.The results from two strategies were compared for every sample.Results A total of 48 HBV full genome sequences were identified into 12 B and 36 C genotype's by both constructing phylogenetic trees and genotyping software analysis,which was exactly the same as the analysis using P gene fragment sequencing.Seven forms of nucleoside drug-resistant mutation were found in the P gene for all the samples,with the ratio of 27.1％ ( 13/48 ),in which all the mutation forms were associated with lamivudine or adefovir,and no other nucleotide drugs-related resistance mutations existed.In addition,there were 11 B and 35 C genotype and 2 B/C hybrid type with the analysis using Real-time PCR genotyping for the 48 samples.Conclusion P gene sequencing can be used as a new clinical method for efficient,accurate HBV genotyping and resistant mutation analysis,which provides guidance for hepatitis B treatment.

Objecflve To evaluate the performance of two rapid and low-cost metheds(MTT test,and rosazurin mierotitre assay)for the detection of resistance to first-line drugs in Mycobacterium tuberculosis.Methods sixty-four Myeobaeterium tuberculosis clinical isohtes were tested by the MTT test and the rosazuxin microtitre assay(REMA)respectively,and the results were compared with those obtained with the absolute concentration method on L(o)wenstein Jensen medium.Results The MTT test and the resazurin microtitre assay showed a good agreement compared with the absolute concentration method for all first-line drugs tested.The sensitibity,specificity and accuracy of the MTT test were 94.8%,96.0%,95.3%,for RFP;93.8%,93.8%,93.8% for INH;92.9%,96.O%,95.3% for EMB,90.6%,87.5%,89.1% for SM,respectively.The sensitivity,specificity and accuracy of the resazurin microtitre assay were 92.3%,96.0%,93.8%,for RFP;90.6%,90.6%,90.6% for INH;92.9%,94.0%,93.8% for EMB,87.5%,87.5%,87.5% for SM,respectively.The Kappa value of the MTT test and the absolute concentration method for the detection of resistance to RFP,INH,EMB,SM were 0.857,0.831,0.714,0.792.respeedvely;The Kappa value of the regazurin mierotitre assay and the absolute concentration method for the detection of resistance to RFP,INH,EMB,SM were 0.871,0.826,0.826,0.750,respectively.The Kappa value of the MTT test and the resazurin microtitre assay for the detection of resistance to RFP,INH,EMB,SM wefe 0.889,0.875.0.787,0.844,respectively.Conclusions Both MTT test and the resazurin microtitre assays are simple,rapid,low-cost and sensitive for rapid detection of resistance to first-line drugs.They could be promising methods for susceptibility assay of the first-line antituberculosis drugs in low-resource countries.

Objective By analyzing the epidemiological characteristics and trends of brucellosis in Ji'nan City,to provide a scientific basis for further prevention and control of the disease.Methods Serum samples of suspected brucellosis patients were collected from every county (city,district) of Ji'nan City from 2008 to 2014,and these samples were screened by Rose-bengal plate agglutination test (RBPT).Then positive samples were confirmed by Standard tube agglutination test (SAT),and experimental data and results were analyzed by statistics.Results Totally 2713 serum samples of suspected brucellosis patients were tested,and 200 were positive by SAT.Temporal distribution:the serum antibody positive specimens were detected each month throughout the year,and the results showed clear seasonal prevalence by concentrating from March to September (82.07％,151/184).Population distribution:the serum antibody positive rate of males was higher than that of females,the ratio was 2.08:1.00,and the cases were found among 15 to 75 years old,while concentrated from 30 to 65 (67.00％,134/200).Spatial distribution:except Huaiyin District,positive samples were detected in all other 10 counties (cities,districts) of Ji'nan City,most of them (92.5％,185/200) in Shanghe,Zhangqiu,Changqing,Shizhong and Licheng counties (cities,districts).Conclusions Brucellosis epidemic in Ji'nan is serious,and cases have occurred mostly in spring,summer and autumn.It is recommended that health propaganda and education should be strengthened in order to enhance selfprotection awareness of the exposed population,and then improve the physical examination for early discovery,early reporting and early treatment.At the same time,it is important to cooperate with the Department of Livestock to prevent and control brucellosis.

Objective To investigate the prevalence of psychological distress and analyze the relevant factors among patients with bladder tumor so as to provide evidence for future clinical practice. Methods Totally 128 patients were recruited from a urological surgery ward of a comprehensive hospital in Beijing in the study by using self-design questionnaire and the psychological distress thermometer (DT) recommended by the U.S. national comprehensive cancer network (NCCN). The acquired data were analyzed by SPSS17.0. Results The average score of the patients was 4.00(1.00~5.00). The identification rate of psychological stress was 55.47%( 71/128 ) , higher than the Chinese normal ( U = 8 . 28 , P < 0 . 05 ) . The relevant factors of psychological stress based on the rank from high to low scores included emotion problems ( 1 . 63 ± 0 . 67 ) , practical problems ( 1 . 42 ± 0 . 64 ) , communication problems (1.29 ± 0.65), physical problems (1.28 ± 0.33) and religion problems (1.00 ± 0.08). Conclusions The prevalence of psychological distress is higher among patients with bladder tumor and the influence factors mainly include emotional problems , practical problems and communication problems. Nurses should pay attention to the psychological distress of patients with bladder tumors and develop targeted interventions so as to relieve their distress.

OBJECTIVE To establish multiple PCR(M-PCR) assay for simultaneous detection of Treponema pallidum(TP) and herpes simplex virus(HSV),Haemophilus ducreyi(HD).METHODS According to specific gene sequence of the three agents:HSV DNA polymerse gene,TP tpp47 gene,HD 6s rRNA gene;three sets of specific primers were designed and a multiple PCR assay was developed to detect 3 agents in one test.RESULTS The target DNA of 3 agents were specifically amplified by their specific primers,the appropriate size of four DNA fragments was 202bp,252bp,152bp for TP,HSV and HD,respectively.The DNA of other several common microbes of genital tract and human genome could not be amplified by three sets of primers.CONCLUSIONS Primary study indicated that the M-PCR assay can simultaneously,specifically and rapidly detect out HSV,TP and HD.

The effect of carbon source,nitrogen source,temperature,moisture,pH and light on the fungal growth,conidia production and conidia germination of Paecilomyces cicadae LB were studied.For the fun-gal growth and conidia production,the optimum carbon sources were soluble starch and glucose,while the optimum nitrogen source was peptone.For the fungal growth and spore germination of P.cicadae,the opti-mum temperature was 25?C~27?C,for the conidia production,the optimum temperature was 25?C.Conidia geminated at RH 90%~100%,but did not below 90%.When RH reaches to 100% or the conidia were in wa-ter,the germination rate was the highest.The range of pH for the fungal growth,conidia production and co-nidial germination was 4~10,while the optimum pH for the fungal growth was 6 and 6~7 for the conidia production and conidial germination.The light treatment significantly influenced fungal conidia production.The lethal temperature for the fungal conidia was 55?C remaining 10 minutes.The present results suggest the isolate LB can adapt to nutrition and environment more widely,and has greater potential as biological control factor against of pest.

Objective To screen primers used in polymerase chain reaction (PCR) for detecting Trichomonas vaginalis. Methods Three pairs of PCR primer reported in the literatures (TVA5-TVA6, TV1-TV2 and TVK3-TVK7) were screened. For each PCR, four components, including primers, Mg2+, dNTPs and Taq polymerase, were optimized using Taguchi methods to determine the optimal PCR conditions. With the optimal conditions, the sensitivities of three PCR were compared. Vaginal swabs were collected to detect Trichomonas vaginalis by culture and PCR, and the PCR with highest sensitivity was evaluated. Results All three PCR were of high specificity, and the PCR with primers of TVK3-TVK7 had the highest sensitivity. Of 25 clinical vaginal swabs, T. vaginalis was detected in 7 samples by the culture, however, it was detected in 8 samples by the PCR. All culture-positive samples were also positive by PCR. Conclusions The PCR with the primers of TVK3-TVK7 is highly sensitive and specific, which could be useful to detect T. vaginalis in vaginal swab samples.

OBJECTIVE:To compare the in vitro antibacterial activity of azlocillin given alone with azlocillin plus tazobactam against168strains of clinically isolated pathogenic bacteria.METHODS:The antibacterial activities of two antibac_ terials against168strains of clinically isolated pathogenic bacteria in vitro were detected by two-fold dilution method.RE-SULTS:The MIC 50 and the MIC 90 of the combined therapy of azlocillin/tazobactam(4∶1)were1/32and1/64,respectively that of azlocillin given alone.CONCLUSION:The concomitant therapy of azlocillin with tazobactam improves the antibacterial activity.

Objective To develop a gene chip for the detection of pathogens causing genital ulcer diseases (GUDs). Methods Specific probes of 4 different pathogens were designed and synthesized. Gene chip was prepared by blotting the probes onto specially treated glass slides with the use of a robotics. Target genes of standard strains for the 4 different pathogens and the clinical specimens were amplified by PCR with Cy5 fluorescence labeled primers. The labeled amplicons were hybridized with gene chips, and then scanned and analyzed using computer software. Results The fluorescence signal for specific pathogen could be found in the gene chip, illustrating that one specific fluorescence signal denoted a single pathogen, and the combination of different signals denoted the corresponding co-existence of pathogens. Examination of 40 clinical specimens obtained from 40 patients with genital ulcers with gene chip was in good concordance with dark-field microscopy plus PCR or HSV culture plus PCR, showing Kappa values of 0.882 and 0.947, respectively. In addition, mixed infections were detected in 2 specimens. Conclusion Gene chip is a sensitive method with a reliable result and it can detect multiple infections simultaneously.