Objective To investigate antimicrobial susceptibility of Klebsiella pneumoniae (K .pneumoniae )to aminoglycosides and detection of 16S rRNA methylase genes in K .pneumoniae .Methods Ninety-six non-repetitive clinical K .pneumoniae isolates were collected from Xiangya hospital of Central South University from January to Ju-ly 2009,minimal inhibitory concentrations (MICs)of gentamycin,amikacin and tobramycin were determined by agar dilution method ;genotype of 16S rRNA methylase genes (armA,rmtA,rmtB ,rmtC,rmtD ,npmA)were detec-ted by polymerase chain reaction(PCR).Results MIC50 of amikacin,gentamycin and tobramycin was 256μg/mL, 512μg/mL and 512μg/mL respectively;and MIC90 were all>512μg/mL;antimicrobial resistance rate was 21 .88%, 63.54%,and 41 ,67% respectively.68 isolates (70.83%)were resistant to at least one kind of antimicrobial agent, 21 isolates(21 .88%)were resistant to three kinds of antimicrobial agents.22 isolates(22.92%)carried armA,but rmtA,rmtB ,rmtC,rmtD and npmA were not detected;of 22 isolates harboring armA 16S rRNA methylase genes, 17(77.27%)were highly resistant to gentamicin,amikacin and tobramycin,the homology of armA positive isolate and armA (FJ410928.1 )was 100%.Conclusion armA 16S rRNA methylase gene harbored in K .pneumoniae plays an important role in aminoglycoside resistance.

Objective To assess the diagnostic value of ChromID for Clostridium Clostridium infection (CDI) on the premise of large sample size,with Meta-analysis.Methods The literature inclusion and exclusion standards were made,and the databases were searched,including PubMed,Cochrane Library,China Biology Medicine disc (CBM),China National Knowledge Infrastructure (CNKI) and so on,using the keywords (ChromID,Clostridium difficile,and chromomgenic agar) along with each of the following words (detection,diagnosis,and infection).Quality assessment of diagnostic accuracy studies (QUADAS) was used to evaluate the quality of the literature.The Meta-disc 1.4 software was used to analyze the data.Results Five studies on 1 851 patients were included in this meta-analysis.The pooled sensitivity was 0.793 (95 ％ CI:0.750 ～ 0.832) for the diagnosis of CDI on 48 h,the pooled specificity was 0.942 (95 ％ CI:0.972 ～0.955),the pooled positive likelihood ratio was 16.817 (95％ CI:7.042 ～ 40.157),with negative hkelihood ratio was 0.124 (95％ CI:0.040 ～0.386) and diagnostic odds ratio for 244.64 (95％ CI:17.711 ～3 379.3).The area under the working curve (AUCSROC) of the subjects was 0.981 4 and the Q* index was 0.939 9.Conclusions Meta-analysis on ChromID for diagnosis of clostridium difficile infection revealedChromID had higher sensitivity,specificity,positive likelihood ratio,and negative likelihood ratio,which can be recommended for clinical use.

Objective To explore the clinical application value of the prealbumin (PA),total bile acid (TBA),and red blood cell volume distribution width (RDW) in chronic liver disease.Methods Totally 393 cases of patients with chronic liver disease admitted by Xiangya Hospital of Central South University from March 2015 to March 2016 were selected as group observation,and were divided into chronic hepatitis,compensated liver cirrhosis,decompensated liver cirrhosis and primary liver cancer.At the same time,200 cases of healthy volunteers were collected as normal control.Serum prealbumin and total bile acids were tested as well as the RDW of all cases.SPSS 17.0 software was used for data statistics processing.The receiver-operating characteristic curve (ROC) was drawn to evaluate the diagnosis value of the indexes in chronic liver disease severity.Results PA in the observation group was significantly lower than normal control,while its TBA and RDW were significantly higher than normal control.All of three parameters in patients,especially with liver cirrhosis and decompensated liver cirrhosis,had higher positive rate.When the clinical diagnosis was taken as gold standard,the best level of PA to diagnose primary liver cancer,chronic hepatitis and decornpensated liver cirrhosis was 244.7 mg/L,238.5 mg/L and 132.8 mg/L,the AUC was 0.973,0.909 and 0.879,the sensitivity was 92.3％,95.1％ and 85.6％,and the specificity was 95.8％,72.8％ and 79.7％;the best level of RDW to diagnose primary liver cancer,chronic hepatitis and decompensated liver cirrhosis was 13.2％,13.8％ and 14.3％,the AUC was 0.816,0.827 and 0.818,the sensitivity was 66.7％,77.4％ and 72.2％,and the specificity was 79.5％,73.8％ and 77.3％.When combined detection of PA and RDW,the diagnostic performance had improved significantly.Conclusions Serum prealbumin and total bile acid,as well as the whole blood RDW may objectively reflect the injury of liver metabolism and synthesis function,and for the early diagnosis and prognosis of patients with chronic liver disease has a important clinical significance.

Objective To determine diazepam, nitrazepam, oxazepam, estazolam, and al-prazolam simultaneously in human plasma by reversed phase high-performance liquid chromatography (RP-HPLC). Methods Ten microliter carbamazepine (50 mg/L)as the internal standard was added into 1 mL sample, which contained the 5 mixed sedative hypnotics as standard substance and human plasma as ground substance. They were extracted with acetoacetate from plasma samples, and then were dissolved by 100 μL mobile phase. The blood drug levels were analyzed by high perform-ance liquid chromatograph with 20 μL sample injection on a chromatographic column C 18 (4.6 mm×250 mm)at 30℃. The mobile phase consisted of methanol and water (65:35) , and the flow rate was 1.0 mL/min. The ultraviolet detection wavelength was 230 nm. Results The linearity range of the 5 drugs was 5～1 200 μg/L(r≥0.9966, P<0.05). The recovery rate was 95.5%～105.6%. The extraction recovery rate was more than 75%. The relative standard deviation (RSD) of intra-day and inter-day was less than 10% (n=5). Conclusion RP-HPLC method is convert-ient, accurate and sensitive for simultaneous determination of the concentration of diazepam, nitraze-pam, oxazepam, estazolam, and alprazolam in human plasma.

Objective To analyze strains of Proteus mirabilis (P .mirabilis )and Proteus vulgaris (P .vulgaris ) isolated in a hospital,detect resistance to commonly used antimicrobial agents,and provide reference for rational ap-plication of antimicrobial agents in clinic.Methods 172 P .mirabilis isolates and 68 P .vulgaris isolates isolated between January 1 ,2011 and June 30,2013 were analyzed,antimicrobial resistance susceptibility testing were per-formed by disk diffusion method,data were analyzed with WHONET 5.4 software.Results P .mirabilis strains were mainly isolated from wound secretion(26.74%),sputum(22.68%)and urine(18.61 %),P .vulgaris were mainly from wound secretion(48.53%),urine(17.65%)and sputum(11 .77%).The resistance rates of P .mirabilis to ampicillin and trimethoprim/sulfamethoxazole were both>45.00%;the resistance rates of P .vulgaris to cefazo-lin and trimethoprim/sulfamethoxazole was 86.76% and 41 .18% respectively;the resistance rates of P .mirabilis and P .vulgaris to piperacillin/tazobactam,cefotaxime,ceftazidime,cefepime,carbapenems (ertapenem and mero-penem)and amikacin were all <20.00%.Conclusion The resistance rates of P .mirabilis and P .vulgaris to pip-eracillin/tazobactam,cefotaxime,ceftazidime,cefepime,ertapenem,meropenem and amikacin are all high,and can be used as the empirical medication for the treatment of related infection.

Objective To determine the serum levels of cross-linked telopoptide of type Ⅰ collagen (ICTP),alkaline phosphatase (ALP) in patients with primary malignant bone tumor,primary benign bone tumor and malignant tumor metastasized to the bone,and to explore the clinical value of ICTP and ALP in identification and diagnosis of bone tumor.Methods Sixteen primary malignant bone tumor patients,16 primary benign bone tumor patients and 18 malignant tumor metastasized to the bone patients in 2012 were studied.Serum ALP was assayed by SFBC rate method and ICTP by enzyme linked immunosorbent assay (EIA).Results The serum levels of ICTP was not significantly different between primary benign bone tumors and normal control group (P ＞ 0.05),but the other between-groups had statistically significant difference (P ＜ 0.05).The serum levels of ALP in malignant tumor metastasized to the bone was significantly higher than the rest of the group (P ＜0.01),but the difference between the remaining groups was not statistically significant (P ＞ 0.05).The area under roc curve (AUC) of ICTP for diagnosis of primary benign bone tumors,malignant tumor metastasized to the bone,primary malignant bone tumor (0.923,0.926,0.874) was higher than the ALP (0.354,0.702,0.865).Conclusions Serum ICTP and ALP were sensitive and convenient biochemical indices which reflected metabolism of patients with bone tumor.Serum ICTP was more specific and sensitive than ALP and they have clinical importance for differential diagnosis as an index of bone tumor.

OBJECTIVE To investigate the drug resistance and the prevalence of inducible clindamycin resistance in clinical isolates of Staphylococcus aureus and evaluate the clinical value of cefoxitin disk diffusion method and oxacillin disk diffusion for detection of meticillin-resistant S.aureus(MRSA).METHODS Bacteria identification and susceptibility test were performed by VITEK-2 system and K-B disk method.The PBP2a was detected by latex agglutination and MRSA was identified by cefoxitin disk diffusion method and oxacillin disk diffusion.The inducible resistance of erythromycin to clindamycin was checked by D-test according to the standards of CLSI(NCCLS).The statistical analysis was performed by WHONET 5.4 and SPSS 13.0 software.RESULTS Resistant rate to penicillin and ampicillin was 98.9% and 100.0%,respectively.Vancomycin-resistant(VRE) or intermediate strains were not found.Of the 93 S.aureus isolates,MRSA and meticillin-sensitive S.aureus(MSSA) were 58(62.4%) and 35(37.6%),respectively.The resistant rate of MRSA to 11 antibiotics was higher than MSSA.The sensitivity and specificity of cefoxitin disk diffusion method were 98.3% and 97.1%,respectively,those of oxacillin disk diffusion were 75.9% and 94.3%.Of the 9 isolates resitant to erythromycin but susceptible to clindamycin,5(55.6%) showed inducible resistance to clindamycin.CONCLUSIONS Resistance of S.aureus is quite serious.Cefoxitin disc diffusion method is a simple and reliable method for the detection of MRSA.The inducible resistance of erythromycin to clindamycin in S aureus should be checked by D-test in clinical microbiology laboratory routinely.

OBJECTIVE To acquire the information about the gene type and epidemic condition of the hospital to provide scientific proof for monitoring and controlling nosocomial infection.METHODS Meticillin-resistant Staphylococcus aureus(MRSA) was identified by its resistance to cefoxitin of disk diffusion and mecA PCR,randomly amplified polymorphic DNA(RAPD) was carried out with the optimization condition.RESULTS The rate of MRSA infection was 72.15% and the main gene type was A in the hospital.CONCLUSIONS The nosocomial infection may exist in the hospital and the hospital must take effective measure to decline nosocomial infection of the MRSA;RAPD is suitable for molecular epidemiology with high powerful discrimination,simplicity and rapidness.

At present, the laser induced breakdown spectroscopy (LIBS) has already become an analysis method with great application value and bright prospects. It has been reported that the LIBS technology had been successfully applied in the field of clinical laboratory diagnostics such as the identification of pathogens, the diagnosis of malignant tumors, the identification of caries, the analysis of onychomycosis, the detection of toxic and harmful elements and other applications with positive results. The remote-LIBS technique provides feasibility for detecting pathogen which is with significance of the disease prevention and control. It is possible to apply LIBS technology to in vivo diagnostics in the future.

Objective To investigate the distribution and change in antimicrobial resistance of pathogens causing blood-stream infection,so as to provide reference for rational antimicrobial use.Methods The isolation and antimicrobial resistance of major pathogens from blood culture specimens from a tertiary first-class hospital in 2012-2015 were analyzed statistically.Results A total of 4 780 isolates were detected,the top five species were Escherichia coli (n = 1 008, 21.09%),Klebsiella pneumoniae (n = 624,13.05%),Acinetobacter baumannii (n = 452,9.46%),Staphylococcus aureus (n=437,9.14%),and Pseudomonas aeruginosa (n=247,5.17%).The percentage of gram-negative bacilli, gram-positive cocci,fungi,and others were 62.05%,29.31%,7.76%,and 0.88% respectively.The resistance rates of Klebsiella pneumoniae to ertapenem and imipenem increased from 4.50% in 2012 to 46.79% and 33.94% in 2015(both P<0.01).The resistance rates of Acinetobacter baumannii to cefepime,ceftazidime,tobramycin,gentamicin,and imipenem were 86.50%,80.56%,78.10%,79.87%,and 84.29% respectively;resistance rates to amikacin in 2012-2015 were 0, 10.22%,39.85%,and 21.30% respectively(P<0.01);resistance rates to minocycline in four years were 0-7.52% (P<0.01 ).Conclusion The main pathogens causing bloodstream infection are gram-negative bacilli,Acinetobacter baumannii is highly resistant to cephalosporins and carbapenems,resistance rates of Klebsiella pneumoniae to carbapenems increased rapidly.Broad-spectrum antimicrobial agents must be used cautiously to reduce the selective pressure of antimicrobial agents.