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Background and Aims A nanometer silicon membrane sandwich cup system was self-designed. It could concentrate the bacilli via 0.45-μm microporous filter membrane and semi-automate the acid-fast bacilli ( AFB) by a bacteria-staining machine. The aim of the study was to assess the clinical value of our self-designed system for diagnosing tuberculosis ( TB). Methods A total of 1993 sputum specimens obtained from patients with confirmed or suspected TB were subjected to direct or concentrated specimens smear at Xiang Ya Hospital, Central South University between May 2012 and February 2013. In addition, all the specimens were also inoculated into Lowenstein- Jensen ( L- J) media, and culture results were considered as the gold standard for calculating sensitivity and specificity. Results Compared with direct smear examination, an increased density of red stained bacilli was observed in the self-designed nanometer silicon membrane sandwich cup analysis under the microscope. The positive rate of the self-designed analysis was significantly higher than that of direct AFB smear [10.9% (217/1993) vs 6.2% (123/1993), P < 0.05]. The sensitivity of the self-designed system increased (97.3% vs 55.2%, P < 0.05) without a loss of specificity (100% vs 100%) for identifying positive TB cases compared with the direct smear method. Conclusion The self-designed nanometer silicon membrane sandwich cup and semi-automatic bacteria-staining machine could more efficiently and rapidly detect the AFB in respiratory specimens than direct microscopy. This is a novel and safe examination, and it may replace direct smear examination for the diagnosis of patients with TB. [ABSTRACT FROM AUTHOR]

Introduction: Clostridium difficile infection (CDI) remains a diagnostic challenge for clinicians. More recently, loop-mediated isothermal amplification (LAMP) has become readily available for the diagnosis of CDI, and many studies have investigated the usefulness of LAMP for rapid and accurate diagnosis of CDI. However, the overall diagnostic accuracy of LAMP for CDI remains unclear. In this meta-analysis, our aim was to establish the overall diagnostic accuracy of LAMP in detection of Clostridium difficile (CD) in stool samples.Material and Methods: A search was done in PubMed, MEDLINE, EMBASE and Cochrane Library databases up to February 2014 to identify published studies that evaluated the diagnostic role of LAMP for CD. Methodological quality was assessed according to the quality assessment for studies of diagnostic accuracy (QUADAS) instrument. The sensitivities (SEN), specificities (SPE), positive likelihood ratio (PLR), negative likelihood ratio (NLR) and diagnostic odds ratio (DOR) were pooled statistically using random effects models. Statistical analysis was performed by employing Meta-Disc 1.4 software. Summary receiver operating characteristic (SROC) curves were used to summarize overall test performance. Funnel plots were used to test the potential publication bias.Result: A total of 9 studies met inclusion criteria for the present meta-analysis. The pooled SEN and SPE for diagnosing CD were 0.93 (95% CI: 0.91-0.95) and 0.98 (95% CI: 0.98-0.99), respectively. The PLR was 47.72 (95% CI: 15.10-150.82), NLR was 0.07 (95% CI: 0.04-0.14) and DOR was 745.19 (95% CI: 229.30-2421.72). The area under the ROC was 0.98. Meta-regression indicated that the total number of samples was a source of heterogeneity for LAMP in detection of CD. The funnel plots suggested no publication bias.Conclusions: The LAMP meets the minimum desirable characteristics of a diagnostic test of SEN, SPE and other measures of accuracy in the diagnosis of CD, and it is suitable as a rapid, effective and reliable stand-alone diagnostic test for diagnosis of CDI, potentially decreasing morbidity and nosocomial spread of CD. [ABSTRACT FROM AUTHOR]

Objective: To investigate the staphylococcal cassette chromosome mec (SCCmec) genotype and molecular epidemiological characteristics of healthcare-associated methicillin-resistant Staphylococcus aureus (HA-MRSA) in a large teaching hospital in China., Methdos: From January 2012 to December 2012, a total of 71 nonduplicate HA-MRSA were collected in a teaching hospital in Changsha, China. SCCmec types were determined by multiplex PCR, and Panton-Valentine leukocidin (PVL) gene was detected by PCR. The homology among the tested isolates was determined using pulsed-field gel electrophoresis (PFGE)., Results: Of the 71 HA-MRSA isolates, 49 (69.0%) carried SCCmec III, 10 (14.1%) carried SCCmec IV, 3 (4.2%) carried SCCmec V and 3 (4.2%) carried SCCmec II; the remaining 6 isolates were not typeable by PCR. Compared with patients having SCCmec I/II/III MRSA infections, those with SCCmec IV/V MRSA infections had a significantly younger age and a similar duration of hospital stay before the first MRSA-positive culture and total hospital stay. PVL genes were strongly associated with SCCmec type IV/V MRSA infections. HA-SCCmec IV/V MRSA strains showed a greater susceptibility to rifampicin, gentamicin, levofloxacin, ciprofloxacin, and tetracycline than HA-SCCmec I/II/III MRSA strains. The 13 HA-SCCmec IV/V MRSA isolates formed one large group at the 55% similarity level. Three PFGE clusters with a similarity index of 85% or more were identified, and unique PFGE profiles were observed in 4 isolates., Conclusion: This is the first report of HA-MRSA isolates carrying SCCmec V in Chinese hospitals. SCCmec types IV and V MRSA clones have emerged in Chinese hospitals, which urges more rigorous surveillance of their spread in healthcare facilities in China.

Aims: To investigate the changes of expression and methylation status of PRDM2, PRDM5, PRDM16 in lung cancer cells after treatment with demethylation agent., Methods: A549 (lung adenocarcinoma cell line), HTB-182 (lung squamous cell carcinoma cell line) and HBE (normal bronchial cell line) were treated with 5-aza-2dC. The methylation state of PRDM2, PRDM5, PRDM16 was detected by MSP. The expression of PRDM2, PRDM5, PRDM16 was detected by RT-PCR and Western blot analysis. Cell growth was detected by MTT assay., Results: 5-aza-2-dC reduced the methylation of PRDM2, PRDM5, PRDM16 gene in A549 and HTB-182 cells but not in HBE cells. Consistently, 5-aza-2dC increased mRNA and protein expression of PRDM2, PRDM5, PRDM16 in A549 and HTB-182 cells but not in HBE cells. Furthermore, 5-aza-2dC inhibited the growth of A549 and HTB-182 cells but not HBE cells., Conclusions: PRDM2, PRDM5, PRDM16 promoters are methylated and their expression is suppressed in lung cancer cells. Demethylation drug 5-aza-2dC could upregulate the expression of PRDM2, PRDM5, PRDM16 and suppress lung cancer cell growth. 5-aza-2dC has potential to be used for lung cancer therapy by epigenetic mechanism.

Objective: To investigate the resistance profiles and the trend of bloodstream-infecting pathogens isolated from hospitalized patients during 2004-2010., Methods: The bloodstream isolates were collected from 18 hospitals in 17 cities. Minimum inhibition concentrations (MIC) were determined using the agar dilution method recommended by CLSI (Clinical and Laboratory Standards Institute), and susceptibility results were analyzed according to the 2011 CLSI guideline., Results: Among the 2004-2005, 2007-2008 and 2009-2010 periods, the proportions of clinical isolates were similar; 43.1% (149 isolates), 34.0% (151 isolates) and 47.5% (776 isolates) for Gram positive strains, 56.9% (197 isolates), 66.0% (293 isolates) and 52.5% (858 isolates) for Gram negative strains, respectively. The isolating rate of MRSA was 54.1% (20/37) in 2007-2008, which was the highest among the 3 periods during 2004 to 2010, while it decreased in 2009-2010 (36.5%, 62/170). The MRCNS proportions were similar across the 3 periods. One (1.8%) vancomycin-resistant Enterococcus faecium and 1 linezolid-resistant Enterococcus faecalis were found. Although the isolating rates of penicillin non-sensitive strains (oral) were similar between 2009-2010 and 2007-2008 [54.5% (6/11) and 53.9% (7/13), respectively], the resistant rates increased from 0% in 2007-2008 to 30.8% (4/13) in 2009-2010. The results were similar according to the non-meningitis criterion (IV), and the susceptibility rates decreased from 100.0% (11 isolates) in 2007-2008 to 84.6% (11/13) in 2009-2010. ESBL-harboring strains in E. coli were similar among the 3 periods during 2004 to 2010 [66.7% (30/45), 73.2% (71/97) and 67.9% (233/343), respectively]. ESBL-producing strains in Klebsilla pnuemoniae decreased year after year, 72.4% (21/29), 50.0% (18/36) and 41.1% (65/158) in 2004-2005, 2007-2008 and 2009-2010, respectively. Except that the sensitive rate of Enterobacter cloacae to ertapenem was 80% (32/40), the sensitive rates of other strains to carbapenems were still above 90% and the resistance rates were less than 5%. Acinetobacter baumannii had the highest multi-drug resistance rate (81.8%, 81/99). One strain (1.0%, 1/99) of Acinetobacter baumannii isolated in 2009-2010 was reported to be pan-resistant., Conclusions: We are facing a more serious situation of bacterial resistance. Acinetobacter baumannii resistance was most serious, usually with the characteristics of multiple drug resistance, and even pan-resistance. Carbapenems remain to be the most effective against enterobacteriaceae. Strains resistant to novel antibiotics (linezolid and tigecycline) have emerged.

Objective: To investigate the pathogen profile of nosocomial infection in China, and to survey the susceptibility rates of these pathogens to the clinical common antibiotics., Methods: The non-repetitive nosocomial pathogens isolated from bloodstream infection (BSI), hospital acquired pneumonia (HAP) and intra-abdominal infection (IAI) and the case data were collected from 13 teaching hospitals in different areas of China and sent to a central laboratory for re-identification and susceptibility testing. The levels of minimal inhibitory concentration (MIC) of the common antibiotics were determined by agar dilution method. The data were analyzed by WHONET 5.6 software., Results: A total of 2103 clinical isolates were collected from January to December 2011, of which gram positive cocci and gram negative organisms accounted for 23.2% and 76.8% respectively. The top three pathogens of BSI were E. coli (31.0%, 243/784), K. pneumoniae (14.8%, 116/784) and S. aureus (10.6%, 83/784). The top three pathogens of HAP were A. baumannii (24.2%, 158/652), P. aeruginosa (23.0%, 150/652) and K. pneumoniae (16.4%, 107/652). The top three pathogens of IAI were E. coli (34.3%, 229/667), E. faecium (13.3%, 89/667) and K. pneumoniae (9.6%, 64/667). Methicillin-resistant S. aureus (MRSA) and coagulase negative Staphylococcus (MRCNS) accounted for 64.4% and 78.1% respectively. The susceptibility rates of Staphylococcus species to tigecycline, vancomycin, teicoplanin and linezolid were all 100%. The prevalence of MRSA in HAP was significantly higher than that in BSI or IAI. The susceptibility rates of Enterococcus species to tigecycline, teicoplanin and linezolid were all 100%. The prevalence of extended-spectrum β-lactamases (ESBL) was 64.3% in E. coli and 38.3% in K. pneumonia. Against Enterobacteriaceae, the most active agents were as following in order: tigecycline (92.3% - 100%) [except P.mirabilis], meropenem (87.5% - 100%), imipenem (87.5% - 100%) [except M. morganii], amikacin (87.5% - 100%), polymyxin B (75% - 100%) [except S. marcescens, P. mirabilis and M morganii], cefepime (67.8% - 100%), cefoperazone-sulbactam (66.6% - 100%), piperacillin-tazobactam (61.5% - 100%). Carbapenem-resistance Enterobacteriaceae strains emerged. The susceptibility rates of P. aeruginosa to imipenem and meropenem were 66.2% and 72.2%, respectively. The susceptibility rates of A. baumannii to imipenem and meropenem were 27.7% and 25.9%, respectively. The most active agents against A. baumannii were polymyxin B (100%), followed by tigecycline (79.8%) and minocycline (50.4%). The susceptibility rates of P.aeruginosa to antibiotics in BSI were higher than those in HAP and IAI. Susceptibility rates of S. maltophilia to trimethoprim-sulfamethoxazole, minocycline and levofloxacin were about 90% or above. Susceptibility rates of B. cepacia to trimethoprim-sulfamethoxazole, ceftazidime and meropenem were all 100%. Several P.aeruginosa and A. baumannii strains were resistant to all tested antibiotics except polymyxin B., Conclusions: The pathogen profile is different in different types of infection. The prevalence of multi-drug resistant A. baumannii is high, which is still a key problem of nosocomial infection. Tigecycline remains relatively high activity against gram-positive cocci and gram-negative bacteria (except P. aeruginosa and P. mirabilis) in vitro.

Objective: To investigate the characteristics of New Delhi metallo-beta-lactamase-1 (NDM-1) gene of Klebsiella pneumoniae (K. pneumoniae) emerging in Hunan, and its relationship to antibiotic resistance., Methods: The clinical strain was isolated from a sputum sample of a child with severe pnemonia and toxic myocarditis who was admitted into a general hospital of Hunan Province. VITEK-2 compact instrument was used for bacterial identification and antibiotic susceptibility test. Modified Hodge test was used for the screening of carbapenemase. EDTA-synergy test and combination disk diffusion test were used for detection of metallo-β-lactamase (MBL). PCR was performed for amplification of NDM-1 genes and the positive products were sequenced and analyzed with BLAST. Conjugation was also performed to analyze mechanisms of antibiotic resistance. The results of antibiotic susceptibility tests were compared before and after conjugation., Results: The isolated strain was identified as K.pneumoniae. Modified Hodge test, EDTA-synergy test and combination disk diffusion test were all positive for the strain. The homology between gene sequence of PCR amplification products and NDM-1 gene FN396876.1 in the GenBank was 100%. Transconjugant DNA was used as template for the amplification of NDM-1 gene. The amplification products were sequenced and found to be the same as the NDM-1 gene amplification product of the donor strain. The MIC of transconjugant E.coli J53 (NDM-1) to all the β-lactams increased significantly compared with the recipient strain E.coli J53. The MIC of ertapenem and imipenem increased by more than 8 times, while the MIC of ceftazidime and ceftriaxone increased by more than 64 times., Conclusions: This study first identified a strain of K. pneumoniae carrying NDM-1 in mainland China. NDM-1 gene can be transmitted among different strains and causes extensively drug-resistance to β-lactams.

Objective: To investigate the relationship between emergence of AmpC enzyme and drug resistance in the gram-negative bacilli in Xiangya Hospital and to provide information for the treatment of antibiotics., Methods: The bacteria were identified; the susceptibility was determined by Microscan microbiological identification system; and the AmpC enzyme was detected by disc diffusion method., Results: Of the 204 gram-negative strains, the positive rate of AmpC enzyme was 19.61%; AmpC-positive strains resisted to ceftriaxone, piperacillin, ceftazidime, cefotaxime, cefoperazone, aztreonam, piperacillin/tazobactam, cefuroxime, ceftazidime/clavulanic, and cefotaxime/clavulanic. But AmpC-negative strains were susceptible to those antibiotics. There were significant differences, between the two groups (All P < 0.05). Both groups were susceptible to cefepime, imipenem, meropenem, cefoperazone/sulbactam, levofloxacin, and amikacin., Conclusion: The detection of AmpC enzyme is helpful to the screen of drug resistant strains. Most bacilli with AmpC enzyme show the resistance to third-generation cephosporins, aztreonam, and the beta-lactamase inhibitors, but susceptible to imipenem, cefepime, meropenem, cefoperazone/sulbactam, levofloxacin, and amikacin, which provides the information for the antibiotic therapy.