Objective To evaluate the relationship between functional oipA gene of Helicobacterpylori and digestive disease. Methods Biopsy gastric mucosa were obtained from 360 patients who would get gastroscopy.Hp were isolated from urease positive samples and partly urease negative samples,and improved by microscope,urease experiment and 16SrRNA PCR. OipA and cagA of the isolates were obtained by PCR and the statas of oipA signal region were analysised after sequencing. Analyze the relevance between functional oipA gene of Helicobacter pylori and digestive disease.Results 106 isolated Hp were obtained,in which 72 strains with cagA gene positive,87 with oipA signal region gene positive (80 the statas of signal region was on and 1 was off;6 could not be decided).The frequency of functional oipA were both significantly higher than cagA in ulcer and atrophic gastritis.Conclusion The relationship between functional oipA gene and digestive disease was more closer than that of cagA.

Objective To establish an indirect ELISA method with Helicobacter pylori (HP)recombinated antigene OipA6 as coating antigen after prokaryotic expression and purification.Methods The ELISA plate was coated with Hp OipA6 recom-binant protein as antigen.Serum specimens and enzyme labeled anti human IgG were added.The reaction conditions was op-timized.The results of indirect ELISA were compared with the detection of oipA gene.Results When Hp OipA6 recombi-nant protein was 1.0 mg/ml,the condition of coating was 2~8℃ one night,the dilution of serum specimens was 1∶100 and enzyme labeled anti human IgG was 1∶4 000,the incubation time of enzyme labeled anti human IgG was 40 min,the chro-mogenic time was 15 min,the result of ELISA was the best.The sensitivity and specificity of the indirect ELISA were both higher than the detection of oipA gene.Conclusion It suggested that the indirect ELISA,in which Hp OipA6 recombinant protein were coated,may be an appropriate assisted method in detection of high virulent Hp infection.

Objective To understand the status of oipA gene of Helicobacter pylori (H.pylori) and to evaluate the relationship between oipA gene and digestive disease, Methods Biopsy gastric mucosa were obtained from 360 patients with digestive disease under gastroscopy. H.pylori was isolated from urease positive and some negative samples, and then identified by microscope, urease test and 16 S rRNA PCR. oipA and cagA genes of the isolates were amplified by PCR and the status of oipA signal region was analyzed after sequencing. The relationship between functional oipA gene of H. pylori and digestive disease was investigated. Results One hundred and six isolats were obtained. Seventy-two strains were positive for cagA gene and 87 were positive for oipA gene in signal region. Eighty strains were on open status in signal region called functioal oipA gene and 1 was on off status. The status of other 6 strains were uncertain. The frequency of functional oipA in ulcer and atrophic gastritis were higher than that of cagA(100% and 78.26% vs 58.33% and 39.13%).Conclusion Functional oipA gene is closely related with ulcer and atrophic gastritis.

Objective:To express Helicobacter pylori oipA gene with different fragments and determine the protein and its peptides with finest antigenicity.Methods:oipA gene was obtained from international standard Hp NCTC 11637 and cloned into vector pGEM-T then was identified by PCR and sequencing.Six different fragments of oipA gene were amplified by PCR with the template pGEM-T/oipA.The gene fragments and expressing vector pET-42a were ligated.The recombinant vector were transformed into BL-21.The correct recombinants were identified by PCR,enzyme digestion and sequencing.Recombination protein and the peptides were detected by Western Breeze chemiluminescent after induced by IPTG.Optimization of the time and IPTG concentration for induction were conducted.The expressed products were affinity purified by Ni-NTA of His?Bind.The antigenicity of the recombination protein was determined with Western blot indicated by goat anti-Hp polyclonal antibody,and the recombination protein with finest antigenicity was selected with indirect ELISA.Results:The six oipA gene fragments could all expressed and the products were recognized by goat-anti Hp polyclonal antibody.The finest peptide with antigenicity was the smallest peptide.Conclusion:OipA gene fragments could be expressed in prokaryotic system and the smallest peptide had the finest antigenicity.It may be used as the reagent to detect corresponding antibody in serum.

Objective To evaluate the capability of three tests used alone or in combination for identification of Staphylococcus aureus.Methods Identification of Staphylococcus aureus by the detection of spa gene with PCR and the Vitek-2 system were selected as the reference methods.Comparison of three phenotypic tests including DNase,mannitol fermentation and tube coagulase test was carried out to analyze the sensitivity,specificity,positive/negative predictive value,positive/negative likelihood ratio and Youden index.The consistency,cost and related indexes of the assays were analyzed between the combined phenotypic tests and the reference methods.Results In the present study,324 isolates of Staphylococci,including 293 Staphylococcus aureus and 31 non-Staphylococcus aureus,were collected.Single biochemical test could not identify Staphylococcus aureus efficiently.Comparison between the reference methods and the combined three biochemical tests by Kappa statistic analysis indicated that an overall Kappa value was 0.9441,and the algorithm of combined test was less costly.The sensitivity and specificity of this algorithm were 100％ and 90.3％,respectively.Conclusion The cost-effective algorithm of combined DNase,mannitol fermentation and tube coagulase test could efficiently distinguish Staphylococcus aureus from other Staphylococcus species.

Objective To investigate the association of genetic polymorphisms in glutathione S-transferases T1 (GSTrl), M1 (GSTM1) and P1 (GSTP1) with aristolochic acid nephropathy (AAN) of Chinese people in Wenzhou of China. Methods Fifty-nine patientswith AAN (AAN group) including 29 male and 30 female as well as 157 healthy ethnically matched controls (control group) including 93 male and 64 female were enrolled in this study. The genotypes of GSTT1, GSTMI and GSTP1 were determined by multiple PCR and confronting two-pair primers PCR (CTPP-PCR). Results The genotype frequencies of GSTP1 were in Hardy-Weinberg equilibrium. Compared with the healthy controls, the frequency of GSTT1 null genotype was significantly higher in the patients with AAN (66.1% vs 48.4%,P＜0.05). Risk of A.AN for individuals with GSTT1 null genotype was 1.747 fold of those without GSTIl null genotype (95% CI=0.818-3.731). The frequency of GSTM1 null genotype, GSTP1 variant genotypes and GSTP1 G allele in the patients and in the controls were 40.7%, 28.8%, 16.1% and 47.8%, 31.8%, 17.5%, respectively, which were not significantly different. No significant differences were found in prevalence of GSTM1 and GSTP1 gene distribution between patients and controls. Conclusion GSTrl gene polymorphism appears to be associated with susceptibility to AAN in Southern China.

Objective To investigate the prevalence and cardiovascular risk factors of silent brain infarct (SBI) in Shunyi Cohort.Methods This study was based on the population based Shunyi Study in China.One thousand and twenty-seven stroke-free participants older than 35 years,who completed cerebral MRI,were included.Cardiovascular risk factors were assessed by interview,physical examination and blood sample tests.SBI was evaluated on 3D-T1WI,T2WI and FLAIR sequences.Associations between risk factors and SBI were analyzed by Logistic regression and adjusted for age,sex,and relevant confounders.Results One thousand and twenty-seven participants,aged (55.9 ± 9.4) years,37.7％ male,were assessed.One hundred sixty-four participants(16.0％) had SBI on MRI.The prevalence of SBI increased by age (every 10 years,OR=2.12,95％ CI 1.74-2.58,P＜0.01).Hypertension(OR =2.67,95％ CI 1.77-4.04,P＜0.01),diabetes(OR=2.48,95％ CI 1.64-3.76,P＜0.01) and smoking(OR=1.98,95％ CI 1.08-3.62,P =0.028) were significantly associated with SBI.Conclusions The prevalence of SBI in this Chinese population is 16.0％,which increases with age.Hypertension,diabetes and smoking are associated with SBI.

Objective:To investigate the clinical effect of hysteroscopic scar excision for cesarean scar pregnancy(CSP) and its effect on serum β-human chorionic gonadotropin(β-HCG).Methods:From January 2017 to January 2019, 83 patients with endogenous CSP admitted to the People's Hospital of Wenzhou were divided into observation group(43 cases) and control group(40 cases) according to different surgical methods.The observation group underwent hysteroscopic scar lesion resection, while the control group underwent uterine artery embolization.The success rate, intraoperative bleeding volume, vaginal bleeding time and hospitalization time, postoperative complications and the recovery time of blood β-HCG were compared between the two groups.Results:The success rate of operation in the observation group was 97.67%(42/43), which was higher than that in the control group[82.50%(33/40)](χ 2=3.875, P<0.05). The amount of bleeding in the observation group[(43.28±10.28)mL] was less than that in the control group[(74.32±15.48)mL], the time of vaginal bleeding in the observation group[(5.79±1.45)d] was shorter than that in the control group[(8.54±1.89)d], and the time of hospitalization in the observation group[(7.32±1.48)d]was shorter than that in the control group[(10.80±1.97)d], the differences were statistically significant( t=10.832, 7.468, 9.140, all P<0.05). There was no statistically significant difference in postoperative complications between the two groups( P>0.05). The recovery time of blood β-HCG in the observation group[(19.54±3.52)d]was shorter than that in the control group[(28.73±6.57)d]( t=8.020, P<0.05). The total satisfaction rate of the observation group was 97.67%(42/43), which was higher than 77.50%(31/40) of the control group(χ 2=6.170, P<0.05). Conclusion:Hysteroscopy-assisted scar excision is effective in the treatment of CSP, and can shorten the recovery time of blood β-HCG.

Objective To investigate antimicrobial resistance among gram-negative bacilli in China in 2010.Methods A total of 1 259 consecutive and non-repetitive gram-negative bacilli were isolated from 13 teaching hospitals from September to December in 2010 in China.All of these isolates were sent to the central laboratory for re-identification and susceptibility testing.The MIC of meropenem and other antibacterial agents were determined by agar dilution method.Interpretive results was determined by CLSI M100-S21.Results The activity of 14 antibacterial agents against 845 Enterobacteriaceae isolates was as follows in order:meropenem ( 98.1％,829 ),amikacin ( 94.0％,794 ),imipeuem ( 90.0％,761 ),piperacillin/tazobactam ( 87.5％,739 ),cefepime ( 83.0％,701 ),ertapenem ( 82.4％,696 ),cefoperazone/sulbactam ( 80.3％,678 ),colistin (75.4％,637),ceftazidime (70.0％,591 ),ciprofloxacin (59.1％,499 ),cefoxitin ( 54.8％,463 ),ceftriaxone ( 53.5％,452 ),cefotaxime ( 52.3％,442 ) and minocycline(51.5％,435).The prevalence of ESBL was 61.3％ (106/173) in Escherichia coli,which was much higher than 41.2％ (70/170) in Klebsiella pneumoniae.The susceptibility rate of Klebsiella pneumoniae against meropenem,imipenem,amikacin and colistin were more than 90％,but were highly resistant to ceftriaxone and cefotaxime.Over 80％ of Enterobacter cloacae,Enterobacter aerogenes,Citrobacter freundii,were susceptible to meropenem,amikacin,cefepine,cefoperazone/sulbactam,imipenem,piperacillin/ tazobactam,and ertapenem.The most active antibiotics against Pseudomonas aeruginosa were Colistin (98.4％,182),Amikacin ( 85.9％,159 ),Piperacillin/Tazobactam ( 80％,148 ),Ceftazidime ( 79.5％,147),Meropenem (74.1％,137),Ciprofloxacin (74.1％,137),Cefepime (73.5％,136),Imipenem (71.9％,132) and Cefoperazone/Sulbactam (70.8％,131 ).Less than 37％ of Acinetobacter baumannii isolates were resistant to carbapenems.The susceptible rate to Minocycline was 47.8％.Colistin kept good activity against Acinetobacter baumannii (susceptible rate,97.8％,n =176),The prevalence of Pan-drug resistant P.aeruginosa and A.baumannii was 18.9％ (n =35),and 61.8％ (n =108),respectively.Conclusions Carbapenems remained very high activity against Enterobacteriaceae.Increasing resistance to the antimicrobials agents test among A.baumanni and P.aeruginosa,especially carbapenems among A.baumanni brought great concern.

Objective Toinvestigateantimicrobialresistanceamonggram-positivecocciinChinain 2013.Methods Retrospectivestudy.FromJune2013toDecember2013,1663consecutiveandnon-repetitive gram-positive cocci were collected from 15 teaching hospitals. The minimal inhibitory concentration ( MIC) of antibacterial agents was determined by agar dilution method. A retrospective study was conducted on rates of resistance to antimicrobial agents. The prevalence of penicillin-resistant Streptococcus pneumoniae ( PRSP) between children and adult patients and the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) between elder group and younger adult patients were compared using chi-square test. Results The prevalence of PRSP in children below 3 years old ( 72. 9%, 51/70 ) was higher than adult patients (55. 2%, 106/192) (χ2 =6. 653,P<0. 05). About 94. 9%(261/275) and (92. 7%,255/275) of S. pneumonia were resistant to erythromycin and clindamycin. All S. pneumoniae strains were susceptible to teicoplanin, vancomycin, linezolid, tigecycline and daptomycin. Penicillin still showed very high activity against Streptococcus spp. β-Hemolytic group. More than 60% of Streptococcus spp.β-Hemolytic group were resistant to erythromycin, clindamycin and tetracyclines. The prevalence of MRSA and methicillin-resistant coagulase-negative Staphylococci(MRCoNS) was 39. 7%(229/576) and 80. 6%(224/278), respectively. The MRSA prevalence ranged from 24. 2% to 70. 0% in different regions. About 52. 6%( 100/190 ) of Staphylococcus aureus from respiratory tract specimens, 38. 5%(40/104)of Staphylococcus aureus from blood samples, and 29. 7%(58/195) of Staphylococcus aureus from wound and pus were resistant to methicillin. The prevalence of MRSA in elder group ( 48. 6%, 84/173 ) was higher than that in younger adult patients (35. 7%, 144/403)(χ2 =8. 322,P <0. 05). The susceptibility rates of MRSA to chloramphenicol and trimethoprim/sulfamethoxazole were 86. 4% ( 244/228 ) and 94. 7% ( 237/228 ) , respectively. Susceptibility rates to gentamycin, erythromycin, clindamycin, tetracyclines, rifampicin and quinolones were ranged from 15. 8% to 59. 6%. All Staphylococci isolates were susceptible to teicoplanin, vancomycin, linezolid, daptomycin and tigecycline. All Enterococcus isolates were susceptible to daptomycin and tigecycline. All E. faecalis ( 158/158 ) and 96. 4% ( 133/138 ) of E. faecium were susceptible to teicoplanin. About 98. 0% ( 150/153 ) of E. faecalis and 97. 1% ( 145/138 ) of E. faecium were susceptible to linezoild. About 45. 8% (70/153) of E. faecalis and 60. 9% (84/138) of E. faecium were resistant to gentamycin with a high concentration. The susceptibility of E. faecalis to all the antibiotics tested exceptchloramphenicolandtetracyclinewashigherthanthatofE.faecium.Conclusions Basedon different age groups and regions, the resistance rates of Gram-positive cocci are different. Teicoplanin, vancomycin, tigecycline, daptomycin, linezolid and tedizolid showed very high activity against Gram-positive cocci. (Chin J Lab Med,2015,38:373-381)