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Objective To investigate the distribution and molecular epidemiology of Candida strains isolated from vulvovaginal candidiasis in Shanghai,and conduct molecular phylogenetic analysis of the strains.Methods Candida pathogens were collected from the patients with vulvovaginal candidiasis during the period from August 2015 to February 2016 in Obstetrics and Gynecology Hospital of Fudan University,Shanghai First Maternity and Infant Hospital,and Intemational Peace Matemity and Child Health Hospital.All the strains were identified and statistically analyzed.ATB FUNGUS 3 kit was used to determine the minimum inhibitory concentration of antifungal agents against these strains in vitro.Molecular typing was conducted using multilocus sequence typing (MLST) method.Phylogenetic analysis was carried out by eBURST and MEGA 6 software.Results Of the 2 185 strains of Candida,1 988 were identified as Candida albicans,149 Candida glabrata,20 Candida tropicalis and 28 other Candida species.Overall,6.5％ of the Candida albicans strains were resistant to fluconazole.Twenty-six diploid sequence types (DSTs) were identified among the 93 strains of Candida albicans analyzed,including 50 fluconazole-susceptible strains with sporadic genotypes,but 43 fluconazole-resistant strains clustered as one clonal complex (CC 69) on the same branch (MLST Clade 1) of phylogenetic tree.Conclusions Candida albicans was the main pathogen of vulvovaginal candidiasis in the three obstetrics & gynecology hospitals in Shanghai,which showed slightly higher resistance to fluconazole.It is necessary to monitor the spread of fluconazole-resistant Candida albicans in these hospitals,especially clonal dissemination of CC 69 clone.
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Objective To investigate the resistance rates of the Candida albicans strains isolated from patients with vulvovaginal candidiasis to 5 antifungal agents and examine the mechanism of azole resistance in these strains.Methods A total of 1 646 C.albicans strains were collected in Obstetrics and Gynecology Hospital of Fudan University from January to December 2015.The resistance rates of these isolates to five antifungal agents were analyzed.Azole-resistant (n=30),dose dependent sensitive (S-DD) (n=13),and susceptible isolates (n=10) were randomly selected from the microbiology laboratories of three obstetrics and gynecology hospitals in Shanghai.The expression levels of drug efflux pump related gene CDR1,CDR2,MDR1 and drug target enzyme gene ERG11 were analyzed by real-time fluorescence quantitative polymerase chain reaction (PCR).At the same time,the ERG11 and ERG3 genes were amplified by PCR and sequenced,and analyzed for resistance-related mutations.Results Of the 1 646 C.albicans strains,5.2％,3.2％,2.5％ and 2.1％ were resistant to itraconazole,voriconazole,fluconazole and 5-fluorocytosine,respectively.All isolates were sensitive to amphotericin B.The expression of ERG11 gene was significantly higher in S-DD group and azole-resistant group than in azole-sensitive group (P＜0.05).The expression of CDR1,CDR2 and MDR1 did not show significant difference among the three groups.There were 13 missense mutations in the ERG11 gene,of which T123I,P98S and Y286D amino acid substitutions were newly discovered.Both T123I and Y132H were identified in 26 resistant isolates,of which 16 gene mutation was detected in two pan-azole-resistant isolates.Conclusions The C.albicans strains isolated from vulvovaginal candidiasis showed higher resistance rates to azole antifumgal agents than that to 5-fluorocytosine and amphotericin B.Mutation and over-expression ofERG11 gene may be one of the prevalent molecular mechanisms underlying azole resistance in C.albicans.were pan-azole-resistant.In addition,the ERG3 heterozygous
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Objective To study the extraction process of kendir leaves in compound kendir leaves tablets. Method The optimum extraction process was selected by the orthogonal design L9(34),with the factors of the concentration of ethanol,the volume of ethanol and the time of extraction,the content of quercetin as the evaluating criteria detected by HPLC. Kromasil C18 column (250 mm?4.6 mm,5 ?m) was used. The mobile phase was acetonitrile-0.5% phosphoric acid (27︰73),detection wavelength was 371 nm,the flow rate was 1.0 mL/min,and the column temperature was 30 ℃. Results The best extraction process of kendir leaves was as follows:85% ethanol,30 times volume of ethanol,extracting for 2 hours in boiling water. Quercetin showed a good linearity in the range of 0.963 0～0.080 3 ?g,r =0.999 8 (n =7). The average recovery of quercetin was 99.46%. Conclusion The extraction process was entirely,and the results of determination are satisfactory.