Fifty clinical Mycoplasma pneumoniae strains were isolated from 370 children with respiratory tract infections. Four strains were susceptible to macrolides, while the other 46 (92%) were macrolide resistant. The molecular mechanism of resistance was shown to be associated with point mutations in 23S rRNA at positions 2063 and 2064.Mycoplasma pneumoniae is a common pathogen found in respiratory tract infections of children and teenagers and is commonly treated with macrolides. In recent years, strains which are resistant to common drugs have been isolated from patients (1,(3)(4)(5)(6)(7)(8). In order to evaluate the prevalence of macrolide resistance, we collected clinical samples during 2003 to 2006, cultured M. pneumoniae isolates, and screened for macrolide drug resistance. We investigated the mechanism of resistance by examining the erythromycin target site in the 23S rRNA gene of these strains.Throat swab specimens were collected from 300 inpatient and 70 outpatient children with respiratory tract infection at the Pediatric Department of Beijing Friendship Hospital, affiliated with Capital Medical University, during June 2003 to June 2006. Modified Hayflick medium was used for the isolation and growth of M. pneumoniae. Nested PCR was carried out to verify the identity of M. pneumoniae, using primers which amplify part of the 16S rRNA gene as described previously (2). The MICs of erythromycin, azithromycin, and josamycin required to inhibit M. pneumoniae growth were determined by the microdilution method (1). A reference strain, FH, was used as a drug-sensitive control. Erythromycin resistance was defined as having a MIC of Ն32 g/ml in accordance with the 2006 standards recommended by the CLSI (formerly NCCLS). To examine the molecular mechanisms of drug resistance, the 23S rRNA gene was amplified by nested PCR and the product was sequenced as described previously (8). The DNA sequences were compared with the sequence of M. pneumoniae M129 (GenBank accession no. X68422).Fifty clinical M. pneumoniae strains (44 of them from inpatients) were isolated from the 370 specimens collected. Four strains were susceptible to macrolides, and the other 46 (92%) strains were macrolide resistant. MICs of resistant strains to erythromycin, azithromycin, and josamycin were higher than that of the reference strain and higher than the CLSI guidelines (especially in the case of erythromycin and azithromycin). Table 1 shows the MIC range, MIC 50 , and MIC 90 of clinical isolate strains and the M. pneumoniae reference strain.The 23S rRNA gene sequences of four susceptible strains and the reference strain FH were identical to that of the M.
A fundamental parameter of polluted insulator online monitoring is the leakage current, which has already been shown to be well-related to the pollution discharge of insulators. In this article, in an effort to quantitatively reflect the discharge intensity and the discharge status by the leakage current, we carried out an experimental study on artificial pollution discharge of insulators. A high-speed photographic apparatus was utilized to capture the entire process of local arcs on a porcelain insulator surface, including the arc generation, the arc development, and the flashover, for which the associated leakage current of insulators was synchronously digitized. A comparative analysis of the relation between the two-dimensional discharge image and the leakage current waveform in the process of arc generation and development shows that if the arc area on the insulator surface is relatively small and the leakage current passes through zero, the arc might completely become extinct, whereas this phenomena will not occur if the arc area is larger. In addition, the amplitude of the discharge arc area is found to be roughly proportional to the square of leakage current over the range of leakage current amplitude from 0 to 150 mA. Our results can provide an important guidance for judgment of the discharge status and the discharge intensity on insulator surfaces using the leakage current of insulators.
Clinical isolates of carbapenem-resistant Klebsiella pneumoniae (K. pneumoniae) strains are being increased worldwide. Five pan-resistant K. pneumoniae strains have been isolated from respiratory and ICU wards in a Chinese hospital, and reveal strong resistance to all β-lactams, fluoroquinolones and aminoglycosides. Totally 27 β-lactamase genes and 2 membrane pore protein (porin) genes in 5 K. pneumoniae strains were screened by polymerase chain reaction (PCR). The results indicated that all of 5 K. pneumoniae strains carried blaTEM-1 and blaDHA-1 genes, as well as base deletion and mutation of OmpK35 or OmpK36 genes. Compared with carbapenem-sensitive isolates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the resistant isolates markedly lacked the protein band of 34–40 kDa, which might be the outer membrane proteins of OmpK36 according to the electrophoresis mobility. In addition, the conjugation test was confirmed that blaDHA-1 mediated by plasmids could be transferred between resistant and sensitive strains. When reserpine (30 μg/mL) and carbonyl cyanide m-chlorophenylhydrazone (CCCP) (50 μg/mL) were added in imipenem and meropenem, the MICs had no change against K. pneumoniae strains. These results suggest that both DHA-1 β-lactamase and loss or deficiency of porin OmpK36 may be the main reason for the cefoxitin and carbapenem resistance in K. pneumoniae strains in our hospital.
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