Objective To study the epidemiological features of Cryptococcus neoformans and Cryptococcus gattii isolated from clinical samples in Shenzhen and to elucidate the distribution of species,varieties,genotypes and mating types within the strains tested.Methods The strains involved in this study were 55 cryptococcal strains isolated from our clinical samples.The canavanine-glycine bromthymolblue (CGB) culture was performed to distinguish Cryptococcus neoformans from Cryptococcus gattii.The genotype was characterized by polymerase chain reaction (PCR) fingerprinting with primer M13.The Cryptococcus gattii species and varieties of grubii and neoformans together with two opposite mating type α and a were identified by PCR with variety-specific and mating type-specific primers.The GEF1-restriction fragment length polymorphism analysis was conducted to simultaneously determine the genotype and mating types of strains tested.The sequence type of IGS1 region was analyzed for the VG Ⅱ genotype.Results Of the 55 tested cryptococcal strains,52 were Cryptococcus neoformans,all of which were var.grubii,genotype VN Ⅰ and mating type α.The remaining 3 strains were Cryptococcus gattii,among which,one was genotype VG Ⅰ and mating type α,and two were genotype VG Ⅱ and mating type α.The two VGⅡ genotype strains belonged to the sequence type Ⅱ.Conclusions The strains belonging to the Cryptococcus neoformans var.grubii,genotype VN Ⅰ and mating type α predominate in causative pathogens of cryptococcosis in Shenzhen.Cryptococcus gattii accounts for minority of the cryptococcal isolates,and the highly pathogenic VG Ⅱ genotypes in foreign countries are also characterized.The sequence types of IGS1 region of the two VG Ⅱ strains are in accord with VG Ⅱb sub-genotype.

Objective To investigate the prevalence and molecular characteristics of the extended -spectrum β-lactamase ( ESBL) and AmpC enzyme-producing Proteus mirabilis ( P.mirabilis) strains isola-ted in Shenzhen People′s Hospital.Methods The production of ESBLs and AmpC enzymes by P.mirabilis isolates were detected by a screening and confirmatory test for ESBLs and AmpC disk test , respectively .The PCR assays followed by DNA sequencing of the products were employed to analyze the multiple genes inclu -ding the ESBLs genes, AmpC genes, insertion sequences (ISs) upstream of the ESBLs or AmpC genes, plasmid -mediated quinolone resistance ( PMQR ) determinants , quinolone resistance-determining region (QRDR) genes , the integrase genes, and class1 integron cassette.The epidemiological analysis of the iso-lates was performed by pulsed field gel electrophoresis .Results There were 130 P.mirabilis clinical iso-lates collected from Shenzhen People′s Hospital in China during the year 2004 to 2010.Among them, 13 isolates (10%) produced ESBLs, that accounted for 0%-9.1%in the year 2004-2009 and up to 29.4%in 2010, and 3 isolates (2.3%) produced AmpC enzymes.The predominant genotype of ESBLs -producing isolateswas b al CTX-M-14(n=7), followed by blaCTX-M-65(n=3), blaCTX-M-55(n=1), blaCTX-M-24(n=1) and blaPER-1 (n =1).The clinical isolate of PER-1-producing P.mirabilis was reported for the first time in China.Twoisolates carried an AmpC β-lactamase gene of blaCMY-2 and one isolate carried an unidentified AmpC gene .ISEcp1 located upstream of blaCTX-M and blaCMY-2 were detected in 91.7% (11/12) of CTX-M-producing isolatesand one CMY-2-producing isolate, respectively.ISPa12 was present upstream of blaPER-1 in one studiedisolate.Approximately 66.7% (10/15) of ESBL and /or AmpC-producing isolates harbored PMQR genes including2 carrying qnrD, 5 carrying aac-Ib-cr and 3 carrying both qnrD and aac-Ib-cr.Twelve ESBL and /orAmpC-producers with high level of resistance to ciprofloxacin carried the similar mutation profiles of S 83I inGyrA, S80I or S80R in ParC and among them, six strains showed E466D mutation in GyrB.Approximately86.7% (13/15) of ESBL and/or AmpC-producing isolates carried class 1 integron.Fourteen PFGE typeswere observed among 15 ESBL and/or AmpC-producers.Conclusion The prevalence of CTX-M β-lactamasesin P.mirabilis isolates contributed to the increased resistance to extended -spectrum cephalosporins.The qnrD and/or aac-Ib-cr genes were detected among the most of ESBL and /or AmpC-producing P.mirabilis clinical isolates.

Objective To analyze the pathogens distribution and drug resistance of nosocomial infection in patients with end-stage diabetic nephropathy .Methods 96 cases of end-stage diabetic nephropathy were randomly selected in our hospital .The speci-mens of urine ,blood and sputum were collected .The pathogens were identified by the drug susceptibility testing .Results The in-fection rate was 27 .08% .A total of 103 strains of pathogens were isolated ,including 15 strains of fungi ,42 strains of gram-negative bacteria and 46 strains of gram-positive bacteria .The drugs susceptibility rates of the fungi to flucytosine ,amphotericin B and flu-conazole were 100% ,and it also showed that the fungi had higher sensitivity to other common antibiotics ;the drug susceptibility rates of gram-positive bacteria to vancomycin and teicoplanin were 100% ,while its drug susceptibility ability to penicillin ,strepto-mycin and others were weak ;the drug susceptibility rates of gram-negative bacteria to piperacillin and imipenem were high ,while its drug susceptibility ability to aztreonam and cephalosporin were weak .Conclusion The low resistance in the patients with end-stage diabetic nephropathy the abuse of clinical antiseptic drugs lead to the dysbacteriosis ,resulting in a significant increase of the inci-dence of nosocomial infection ,so the analysis of pathogens distribution and drug resistance of nosocomial infection has clinical sig-nificance .