Objective To investigate acquired carbapenemases and prevalence of carbapenem- resistant gram-negative bacill.Methods The antimicrobial susceptibility was determined by agar dilution method.Metallo-B-lactamase(MBLs)were screened by EDTA-disk synergy tesL The encoding genes of MBLs were amplified by PCR followed by sequencing.Strain homology Was investigated by pulsed-field gel electronphoresis(PFGE).Results In 141 carbapenem-resistant Pseudomonas aeruginosa(P.aeruginosa), there were three resistant patterns which were imipenem(IMP)-resistant+meropenem(MEM)-resistant (66.7%),IMP-resistant+MEM-sensitive(32.6%),and IMP-resistant+MEM-sensitive(0.7%).AⅡthe carbapenem-resistan Acinetobacter baumannii(A.baumannii),Acinetobacter lwoffi(A.lwoffi),Citrobactor freundii(C.freundii),Klebsielta pneumoniac(K.pneumoniac)and Serratia were resistant to imipenem and meropenem.Four strains of 141 P.aeruginosa were positive by EDTA-disk synergy test,and they produced VIM-2-type Metallo-B-1actamase.Of 34 carbapenem-resistan A.baumannii,30 strains produced OXA-tllrpe.Among them, OXA23, OXA24 and OXA66 accounted for 79.4%,38.2% and 67.6%,respectively.And 22 of 34 strains(64.7%)produced multiple OXA-carbapenemases.All 7 strains A.lwoffi produced OXA-23-type carbapenemases.A11 11 strains C.freundii,5 strains k pneumoniac and 1 strains Serratia produced KPC-2-type carbapencmases.And 6 of 11 strains C.freundii produced new subtype IMP-8.Of 15 PFGE type in 34 strains A.baumannii,14 strains belonged to A-type,7 strains belonged to B-type.Seven A.lwoffi strains distilbuted in difierent PFGE type.Four strains of P.aeruginosa producing VIM-2.type Metallo-8-lactamase did not have the same PFGE type.Eleven C.freundii strains had the same PFGE type.Five k pneumoniae strains had the sanle PFGE type.Conclusions Drug resistance to 12 common antibiotics in carbapenem-resistant gram-negative bacill was higher than non-carbapenem-resistant gram-negative.The former produced many kinds of carbaponemases and the strains prducing carbapenemases were prevalent in the C.freundii, A.baumanii, and k pneumoniae.

Objective To analyze the 23S rRNA gene partial sequences of common bacteria, and establish molecular biologic techniques to identify bacteria by the difference of gene sequences. Methods Analyzing the sequences of variable region of bacterial 23S rRNA genes, primers and oligonucleotide probes were designed accordingly. Thereafter, bacteria were identified by PCR gel electrophoresis and PCR reverse hybridization. Results There exists significant sequence difference between Gram negative bacteria and Gram positive bacteria and it could be used to differentiate these 2 kinds of bacteria quickly with PCR gel electrophoresis. Meanwhile, sequence variety in different species of bacteria was also observed and PCR reverse hybridization could be used to identify different bacterial species further.Conclusions There exist significant sequence differences among 23S rRNA genes in different common bacteria. By the sequence differences, a specific, sensitive and rapid molecular biologic techniques could be established to quickly identify the pathogens of bacterial infections.

Objective To evaluate the nursing effect of wenjing moxibustion with ginger in zhuang medicine to knee osteoarthritis (KOA), which can provide an effective nursing intervention for KOA. Methods Totally 80 cases were divided into two groups by random number table. There were 40 cases in zhuang moxibution group and 40 in traditional moxibution group. These two groups were taken the same nursing intervention and exercise. However zhuang group were spend 20 minutes and covered by ginger mud on zhuang xiguan point, and fire moxa on the ginger mud. Xuehai, Liangqiu, Neixiyan, and Waixiyan were chosen and fired moxa cone with ginger in traditional group that spent 20 minutes. It was observed the knee pain and swelling before and after 1 week during the treatment. 2 groups were compared with the cost of treatment. Results There was no different in pain and swilling before treatment (P>0.05), but there was significantly difference after 1 week in two groups (t=14.72、12.90;7.04, 2.73, P﹤0.01). It was significantly different in two groups after 1 week treatment (t=-5.06,-3.64, P﹤0.01). There was no significantly difference in the cost of treatment (F=0.041 6, P>0.05). Conclusions these two nursing intervention can release pain and swelling. However it is effective and no different in the cost of treatment when using wenjing moxibustion with ginger in zhuang medicine.

Some techniques of compounding bioactive ceramics and polymer biomaterials with mechanical and biological properties and the clinical applications of the composites produced are presented.
Biocompatible Materials ; chemistry ; Biomechanical Phenomena ; Ceramics ; chemistry ; Humans ; Polymers ; chemistry ; Surface Properties

Objective To investigate the molecular characteristic,the virulence factors and antimicrobial resistance of Methicillin-resistant Staphylococcus aureus (MRSA) isolated from pediatric patients.Methods Ninety-eight non-duplicate strains of and 49 non-duplicate strains of Methicillinsusceptible Staphylococcus aureus (MSSA) isolated from the three children's hospitals in Shanghai in 2008 were investigated.Panton-valentine leukocidin (PVL) gene was detected by polymerase chain reaction (PCR).The genotypes of staphylococcal cassette chromosome mec (SCCmec) of the MRSA isolates were confirmed by multiplex PCR.The sequence type (ST) of each strain was determined by multilocus sequence typing (MLST),and the algorithm eBURST was used to identify groups of clonal complex (CC).The minimal inhibitory concentrations (MIC) of fourteen antibiotics for all isolates were determined by agar dilution method.Results Among 98 isolates of MRSA,the positive rate of PVL genes was 6.1％ (6/98).In contrast,the positive rate of PVL genes was 4.1％ (2/48) of the MSSA strains.Among 98 isolates of MRSA,4.1％ (4/98),23.5％ (23/98),53.0％ (52/98) and 15.3％ (15/98) of the strains harboured SCCmec types Ⅱ,Ⅲ,Ⅳ and Ⅴ,respectively. The remaining four isolates (4.1 ％) presented a unique SCCmec pattern that could not be classified to any known types by the employed typing assays.Combining the ST and SCCmec type,the predominant clones were ST59-SCCmec Ⅳ (30 strains) and ST239-SCCmec Ⅲ (23 strains),followed by ST5-SCCmecⅣ and ST1-SCCmecⅣ (8 strains for each clone),ST239-SCCmec Ⅴ (6 strains),ST88-SCCmecⅤ (5 strains),ST5 SCCmecⅡ (4 strains),ST59-SCCmec Ⅴ (3 strains),ST8-SCCmecⅣ and ST88-SCCmecⅣ (2 strains for each clone),ST22-SCCmecⅣ,ST910-SCCmecⅣ and S45-SCCmec Ⅴ (1 strain for each clone),eBURST analysis distributed the MRSA isolates into several CC.ST8 and ST239 belonged to ST8 CC,ST1 belonged to ST15 CC,ST910 belonged to ST 30 CC,ST59,ST5,ST88,ST45,ST22,ST9 and ST7 were the origin of their own CC.The results of MIC showed that the 67 strains of MRSA harboring SCCmec type Ⅳ or SCCmec type Ⅴ were more susceptible to various non-β-lactam antibiotics than 27 strains of MRSA harboring SCCmec type Ⅱ or SCCmec type Ⅲ,and no vancomycin-resistant strain was found.Conclusions In three children's hospitals in Shanghai,the PVL gene-positive rate of MRSA isolates is relatively low,SCCmec type Ⅳ and SCCmec type Ⅴ could spread among hospitals to cause a small scale epidemic and have a variety of ST.

Objective To develop a method for rapid detection of Mycoplasma pneumoniae and its macrolide resistance mutation. Methods The primers and cycling probe sets were designed to detect two single nucleotide mutation, A2063G and A2064G, in the 23s rRNA gene of Mycoplasma pneumoniae. By using recombinant plasmids containing 23s rRNA gene fragments, 102 Mycoplasma pneumoniae clinical isolates from 2005 to 2008, and 136 nasopharyngeal suction specimens from pediatric patients with low respiratory tract infections in Shanghai Children's Hospital from November to December in 2009 were investigated to determine the specificity and the sensitivity of the CycleavePCR method. PCR amplification and sequence analysis of 23S rRNA genes were performed for all Mycoplasma pneumoniae strains and Mycoplasma pneumoniae positive specimens to confirm the results of the CycleavePCR method. Results Of 102 clinical isolates, 83 was resistant to erythromycin and sequence results show that all macrolide-resistant Mycoplasma pneumoniae strains harbored an A2063G ( 82/83 ) or A2064G ( 1/83 ) transition mutation in 23S rRNA genes. Twelve was Mycoplasma pneumoniae detected positive by CycleavePCR in 136nasopharyngeal suction specimens. The CycleavePCR results were consistent with those of routine PCR and sequencing. There was no signal production from other bacterial species. Sensitivity and specificity were 100%. The detection limit of the CycleavePCR was 10 plasmid copies per reaction. Experiment can be done within 1.5 h. Conclusion A novel method is developed to detect erythromycin-resistant strains harboring A2063G and A2064G transition mutation in the 23s rRNA gene using CycleavePCR.

Objective To learn the current in vitro antimicrobial susceptibility of Mycoplasma pneu-moniae in Shanghai and to understand the mechanisms of resistance to macrolides. Methods M. pneumoniae was isolated from pediatric patients with low respiratory tract infections(RTI) using broth and PPLO agar medi-um. PCR amplification and sequence analysis of P1 adhesion gene were performed to identify all M. pneumoniae strains. Susceptibility testing was carried out for macrolides, tetracyclines and fluoroquinolones using broth mi-crodilution method with SP4 broth. PCR amplification and sequence analysis of 23S rRNA genes were performed for all M. pneumoniae strains. P1 gene PCR-RFLP typing was performed to subtype the M. pneumoniae strains. Results One hundred and two M. pneumoniae strains were isolated in Shanghai from Oct 2005 to Dec 2008. All M. pneumoniae isolates were susceptible to the tetracyclines and fluoroquinolones tested. Of 102 clinical isolates, 83(81.4%) was resistant to erytbromycin and all 83 erythromycin-resistant strains had MIC>128 mg/L. An increasing trend of resistance rates were showed: 16.7% (1/6) in 2005, 76.5% (13/17) in 2006, 100.0% (24/24) in 2007 and 81.8% (45/55) in 2008. All macrolide-resistant M. pneumoniae strains harbored an A2063G transition mutation in domain V of 23S rRNA genes. The P1 gene RFLP type 1 is predominant (85.3%, 87/102) in M. pneumoniae clinical isolates. Conclusion The macrolide resistance rate of M. pneu-moniae is very high in Shanghai. The mechanism of macrolide resistance is associated with transition mutation on the 23S rRNA gene.

Objective. To understand drug susceptibilities to common antibacterials, resistance mechanism to β-lactams and quinolones and the clonal spread of resistant stains of Haemophilus influenzae (H. influenzae) and Haernophilus parainfluenzae (H. parainfluenzae) isolated from some hospitals in Shanghai. Methods The in vitro antimicrobial susceptibilities to 13 antibacterials, such as ampicillin, of 156 Haemophilus strains collected from 5 hospitals of Shanghai in 2006 were tested by agar dilution method. The β-lactamase production was determined by chromogenic cephalosporin test. TEM and ROB type of β-lactamase genes and quinolone resistance determining regions (QRDR) of ciprofloxacin-resistant strains were detected by polymerase chain reaction (PCR) amplification. The homology of H. influenzae strains were analyzed by enterobacterial repetitive intergenic consensus (ERIC)-PCR. Results The susceptible rate of 109 strains H. influenzae to ampicillin was 74.3%, while those to ampicillin-sulbactam, cephatosporins and fluoroquinolones were all 100.0%. The β-lactamases-producing rates of 109 strains H. influenzae and 47 strains H. parainfluenzae were 25.7% and 19.1% (χ2=0.776,P=0.378), respectively. TEM gene was detected in all β-lactamases-producing strains. Of 109 H. influenzae isolates, only one was resistant to ciprofloxacin, and Ser84Leu mutation was detected in gyrA gene and Gly206Arg mutation in parC gene. The results of ERIC-PCR showed that 106 H. influenzae strains were clustered into 73 groups with similarity level of 85%. Conclusions Clinical isolates of H. influenzae from hospitals in Shanghai remain highly susceptible to common antimicrobial agents except ampicillin. TEM type of β-lactamase production is the main ampicillin-resistant mechanism of the tested stains. The clonal spread of H. influenzae, including ampicillin-resistant strains, is not prevalent.

Objective To investigate thein vitro activity of tigecycline and minocycline against VanM-type vancomycin-resistant Enterococcusfaecium.Methods A total of 45 strains of VanM-type vancomycin-resistantE. faecium were obtained from hospitals in Shanghai between 2006 and 2014. Species and vancomycin resistance genotype were identiifed by PCR and sequencing analysis. Minimal inhibitory concentrations (MICs) of ten antimicrobial agents against these strains were determined.Results All the 45 isolates were VanM-type vancomycin-resistantE. faecium, and resistant to vancomycin (MIC: 128 to >256 mg/L). And 71.1%of the strains were resistant to teicoplanin. Almost all isolates showed resistance to levolfoxacin (100%) and ampicillin (97.8%). About 15.6%, 64.4% and 82.2% of the strains were resistant to minocycline, gentamicin and rifampicin, respectively.Conclusion Tigecycline and minocycline exhibit excellentin vitro activity against these VanM-type vancomycin-resistantE. faecium isolates.

Objective To understand the resistance mechanism and clinical feature of linezolid-resistant S.capitis isolated from blood samples.Methods Antimicrobial susceptibility testing was carried out to determine the susceptibility of clinical strains.PCR and sequencing analysis were used to analyze cfr gene and 23S rRNA mutation,which were associated with linezolid resistance.Patterns of pulsed-field gel electrophoresis (PFGE) were analyzed in combination with clinical data to understand the clinical feature of S.capitis strains.Results Five linezolid-resistant S.capitis strains were isolated from blood samples of 3 patients.These strains were resistant not only to linezolid,but also to most of the commonly used antimicrobial agents except glycopeptides,rifampin,and trimethoprim-sulfamethoxazole.Mutation was identified in 23S rRNA genes of all the five strains and cfr gene was found in four of the five strains.PFGE typing showed the same type,which supported the homology of the 5 strains.Three patients had deep vein indwelling catheter and two of them were treated with linezolid.Conclusions Linezolid-resistant S.capitis isolates showed the phenotype of resistance to multiple antimicrobial agents.Linezolid resistance may be mediated by cfr gene and 23S rRNA mutations in S.capitis.Long-term use of deep vein indwelling catheter and linezolid treatment may increase the risk of linezolid-resistant S.capitis infection.