Objective To study the mechanism of the resistance in Acinetobacter baumannii to common antibiotics.Methods Bacterial susceptibility test to ?-lactamatic antibiotics,quinolones,aminogiycosids were done by the Kirby-Bauer method and agar dilution method for 35 isolates.Penicillin-beta-lactamase,AmpCs,Metallo-beta-lactamase,ESBLs were detected by iodine-starch test,3-dimension test,microbiology sensitivity synergic test,disc agar diffusion method respectively.Outer membrane protein was analyzed by SDS-PAGE.Accumulation of ciprofloxacin was determined by direct Fluorescence method.The gene of Tem-1,aac-4 and gyrA were amplified by PCR while the gyrA was sequenced.Results 28 isolates were multi-resistant to common antibiotics in 35 isolates.16 isolates produced Penicillinase,10 isolates produced Cephalosporinase,2 isolates produced metal-beta-lactamase,3 isolates produced ESBLs.The analysis of outer membrane proteins showed that a protein of 29kD disappeared and 26kD protein enhanced in resistant isolates.The accumulation of ciprofloxacin in resistant isolates decreased.After treatment with NaN_3,the drug uptake increased to the normal level.Most of [QX(Y8]Tem-1 gene were positive except 2 drug resistant and 3 sensitive isolates.All of [QX(Y8]aac-4 were negative while gyrA were positive.DNA sequencing analysis revealed there had point mutation in the gyrA gene.Conclusion Beta-lactamase,active drug efflux,outer membrane protein permeability decreasement and gene mutation were the factors contributing to the antibiotics resistance of Abaumannii.

Objective To analyze the clinical value of sTNFR-1,sIL-2Ra and IGF-1 on depression in patients with esophageal cancer by comprehensive nursing intervention and explore the possibility of serum indicators to replace DSI and HAMD. Methods 120 patients with pathologically confirmed esophageal cancer were included in the study.Zung depression status inventory (DSI)and hamilton depression rating scale (HAMD)were used to assess depression.Serum sTNFR-1,sIL-2Ra and IGF-1were detected by enzyme-linked immunosorbent assay (ELISA).Results 69 patients were classified as anxiety and depression group,51 patients were as normal control group.There were 32 patients with depression.Serum sTNFR-1, sIL-2Ra and IGF-1 levels in study group was higher than that in the control group (P<0.05)and these serological indexs increased from the minimal depression,mild-to-moderate depression group to severe depression group (P<0.05).Conclusion Serum sTNFR-1,sIL-2Ra and IGF-1 levels may be used as evaluation index of nursing intervention,but need further clini-cal validation.

Objective To explore a rapid method for classification of microorganisms.Methods The electrophorese fingerprinting, direct sequence of 16S rDNA and 16S-23S rDNA ISR after PCR, multiplex PCR for 16S rDNA and antibiotic resistance genes, were utilized to explore fast approaches of extracting total DNA from different clinical specimens.Results The specific 16S-23S rDNA ISR fingerprinting fragments were shown on the genus or species level in bacteria and fungi.So fingerprinting can be used to identify pathogenic microorganisms, to differentiate the evolution relations or to set the phylogenetic tree by comparing their DNA banding patterns with those of standard strains (NCCLS). Multiplex PCR was able to examine the special genes of genus or species, mecA gene, TEM, SHV and CTX gene in staphylococcus and ESBLs(E.coli or K.pneumoniae) at the same time.Conclusion The part of 16S rDNA sequencing and 16S-23S rDNA ISR genotypes by gel electrophoreses were useful for bacterial species identification in addition, it was clearly more rapid and accurate than culture technique, and the large numbers of strains can easily be examined.Multiplex PCR could provide a good method for identification of microorganisms and analysis of antibiotic resistance at the same time.

Objective To investigate the regulatory effect of CsA on the expression of NK cell inhibitory receptor ILT4 and cytotoxicity of NK cells.Methods NK cells treated with CsA ( 10 mg/L) or DMSO for 12,24 and 36 h were chosen as three experimental groups and control groups respectively.RTqPCR and flow cytometry were performed to detect the alteration of ILT4 at the mRNA and protein level respectively.The expression of HLA-G in human gastric cancer cell line BGC-823 and human placental choriocarcinoma cell line JEG-3 were measured at the same time,and then the cytolytic activity of the untreated NK cells and NK cells treated with CsA for 36 h against BGC-823 and JEG-3 cells was determined with MTT.One-way analysis of variance was employed to compare the different ILT4 expression at different time points after medication; Dunnett test was performed to carry out the pairwise comparison between each mean.The difference of HLA-G expression between JEG-3 cells and BGC-823 cells,and the difference of NK cell cytolytic activity against JEG-3 cells and BGC-823 cells were analyzed by student's t-test.Results RT-qPCR assay indicated that the relative levels of ILT4 mRNA in NK cells treated with CsA for 12,24 and 36 h in turn were 0.99 ± 0.27,1.79 ± 0.29,6.79 ± 0.64,and those of their contrast groups treated with DMSO were 0.86 ±0.11,0.94 ±0.12,1.06 ±0.17.The expression of ILT4 in NK cells treated with CsA for 24 h or 36 h was higher than that in NK cells of their contrast groups respectively ( t value of 4.69,14.99,P ＜0.05,respectively),but there was no significant difference between the two groups of NK cells treated for 12 h ( t =0.78,P ＞0.05 ).Through flow cytometry,the positive rates of ILT4 protein expression in NK cells treated with CsA for 12,24 and 36 h [(5.16 ± 0.42 ) ％,( 6.23 ± 0.48 ) ％,( 23.8 ± 1.5 ) ％]were higher than those in NK cells after treatment with DMSO for 12,24 and 36 h respectively[(3.08 ±0.19)％,(3.35 ±0.12)％,(3.36 ±0.21 )％ ;t value of 7.70,10.06,20.72,P＜0.01,respectively].The expression of ILT4 in NK cells treated for 36 h was much higher than that in NK cells for 12 and 24 h at the mRNA and protein level (t value of 16.38,14.12 ;21.81,20.56,P ＜ 0.01,respectively).Meanwhile the killing rates of NK cells treated with 10∶1 effector-target ratio CsA on BGC-823 cells (low HLA-G expression) were ( 8 1.96 ± 2.80 ) ％ ( before treatment) and ( 60.23 ± 1.57 ) ％ ( after treatment),which were higher than those on JEG-3 cells (HLA-G-overexpression) [(53.46 ±2.21 )％ ( before treatment),(28.30 ± 1.85 ) ％ ( after treatment)].The changes of cytotoxicity of NK cells treated with CsA against target cells showed that CsA inhibited the killing activity of NK cells to BGC-823 and JEG-3 cells (t value of 11.74,15.16,P＜0.01,respectively),and the inhibitory rates were (26.48 ±2.42)％ and (47.10 ±1.59 ) ％ respectively.CsA had a higher killing rate inhibition on JEG-3 than on BGC-823 ( t =12.31,P ＜0.01 ).Conclusion CsA induces upregulation of ILT4 in NK cells,and the cytotoxicity of NK cells to tumor cells can be affected by interaction of ILT4 and HLA-G.

Objective To establish normal reference interval for four items of blood coagulation on ACL‐TOP Automatic coagu‐lation analyzer .Methods The fasting anti‐coagulation blood samples were collected from 1 268 inpatients and people conducted physical examination ,all subjects without liver disease ,history of blood disease and coagulation disfunction .The prothrombin time (PT) ,activated partical prothrombin time(APTT) ,thrombin time(TT) and serum levels of fibrinogen(FIB) were determined by u‐sing ACL‐TOP automatic coagulation analyzer which was producted by America IL company .And data of determination results were used to establish the normal reference intervals of indexes in this laboratory .Results The precision and accuracy of this analy‐zer was good .There were differences of normal reference intervals between which established in this laboratory and which provided by the manufacturer .Conclusion Each laboratory should establish its own normal reference interval ,not blindly refer to reference interval provided by regents manual .

Objective To investigate the changes of plasma microRNA-223(miR-223) and HMGB-1 in pediatric sepsis patients.Methods There were 49 children with sepsis enrolled in the study (sepsis group),severe sepsis group (n=25) and general group (n=24). Meanwhile, 50 healthy children (normal control group) were selected as control group. The expression levels of plasma miR-223and HMGB-1 (high mobility group box 1) were detected. The predictive values of miR-223and HMGB-1 in plasma of children with sepsis were evaluated by receiver operatingcharacteristic (ROC) curve.Results The plasma miR-223 and HMGB-1 expression levels in severe sepsis group and general group were up-regulated compared with those in the normal control group (F=63.02, 76.32,P<0.05). The area under ROC curve of miR-223,HMGB-1 predicting sepsis were 0.904 (95%CI 0.821-0.998), 0.748 (95%CI: 0.625-0.903). There was positive correlation between miR-223 and HMGB-1 (r=3.532, P<0.05). Conclusions The expression levels of plasma miR-223 in children with sepsis are signiifcantly up-regulated, which can be used as early diagnostic markers to relfect the severity of inlfammation in some degree.

Objective To study the epidemiology and genotypes of extended-spectrum beta-lactamases (ESBL)-producing Escherichia coli (EC) and Klebsiella pneumoniae (KP) that caused community-onset bloodstream infections (COBSI) in 9 county hospitals of Zhejiang Province.Methods This is a multi-center, prospective, observational study.The cases and isolates with COBSI caused by EC and KP were consecutively collected from 9 county hospitals in Zhejiang Province between 1st March 2014 and 30th April 2015.The double disk diffusion method was used to confirm the production of ESBL.The ESBL genotypes were determined by polymerase chain reaction(PCR) amplification and sequence analysis.Multi-locus Sequence Typing (MLST) was used to analyze the homology of ESBL-producing isolates.Minimal inhibitory concentration (MIC) of frequently used drugs for ESBL-producing isolates was determined by in-vitro antimicrobial susceptibility tests.Results During the study period, a total of 172 cases with COBSI were collected and 171 cases were eligible, among which 126 were caused by EC and 45 were caused by KP.The overall prevalence of ESBL was 34.5% (59/171),and the prevalence of ESBL-EC and-KP was 41.3% (52/126) and 15.6% (7/45), respectively.CTX-M-type ESBL accounted for 96.6% (57/59) of all the ESBLs-producing isolates, and the most common type was CTX-M-14 (27.1%, 16/59), followed by CTX-M-55 (22%, 13/59).MLST analyses revealed significant genetic diversity among ESBL-EC and-KP.The most prevalent ST of ESBL-EC was ST131 (23.1%).In addition to carbapenems, cefoperazone/sulbactam, piperacillin/tazobactam, moxalactam, amikacin and fosfomycin also showed good in-vitro activity against ESBL-EC and-KP.Conclusions The prevalence of ESBL in EC and KP is high in 9 county hospitals of Zhejiang Province, and the most common genotypes are CTX-M-14 and CTX-M-55.The detection rate of ESBL in EC is higher than in KP.It could be considered adequate empirical therapy according to the results of antimicrobial susceptibility tests.Carbapenems, cefoperazone/sulbactam, piperacillin/tazobactam, moxalactam, amikacin and fosfomycin have good in-vitro activity against ESBL-EC and-KP.

Objective To explore the value of contrast-enhanced ultrasound in the diagnosis and classification of traumatic spleen rupture , as compared with enhanced CT. Methods The manifestations of contrast-enhanced ultrasound on surgically or clinically confirmed spleen rupture in 40 patients were retrospectively analyzed. The performance of contrast-enhanced ultrasound on diagnosis and classification was compared with that of enhanced CT. Results For 40 patients with traumatic spleen rupture , the of accuracy of enhanced CT and contrast-enhanced imaging in the diagnosis of traumatic spleen rupture was 97.5%, with no significant statistical differences (chi-square = 0, P = 1). On contrast-enhanced ultrasound examination, 14 patients were diagnosed as true splenic rupture , 9 as subcapsular spleen rupture , and 16 as central splenicrupture, with a accuracy rate of 92.5% (37/40); and accuracy rate for enhanced CT was 90.0% (36/40), there was no significant statistical difference (chi-square = 1.97, P > 0.05). Conclusions Contrast-enhanced and enhanced CT have good consistency in the diagnosis and classification of traumatic spleen rupture. Contrast-enhanced ultrasound can accurately determine the scope and degree of spleen damage , resulting in more accurate classification; and it has values in the diagnosis of traumatic spleen rupture and choice of therapies.

Objective To study on virulence characteristics and multilocus sequence type of Vibrio parahaemolyticus isolated from clinic in Beijing Tongren hospital.Methods Total 152 strains of Vibrio parahaemolyticus isolates were collected from diarrheal patients of outpatients in Beijing Tongren Hospital,Capital Medical University from 2009 to 2011.PCR was used to detect hemolysin gene thermo stable direct themolysin gene (tdh),TDH-related hemolysin gene (trh),type Ⅲ secretion system 2 (T3SS2α,T3SS2β)and systematic functional gene (toxRS/new,orf8) for pandemic 03∶ K6 clone and its derivatives.The genetic features of these strains were determined by multilocus sequence typing (MLST).Results 96％ (146/152) VP harbored tdh gene,2％ (3/152) VP harbored trh gene and 100％ (152/152) VP harbored T3SS2 gene.In this study there were 107 pandemic strains (both tdh and toxRS/new positive and trh negative),38 pathogenic strains (tdh positive and/or trh positive) and 6 nonpathogenic strains (both tdh and trh negative).All nonpathogenic strains harbored systematic functional gene (toxRS/new,orf8).Only one pathogenic strains harbored orf8 gene.One clone harbored all virulence gene.In this study there were 16 sequence types,and ST3 is the pandemic sequence type,including 113 strains,and four new sequence types were found.Conclusions In this study more than 90％ Vibrio haemolyticus harbored tdh gene and ST3 was the pandemic sequence type in Beijing.One can get bacterial pathogenic charateristic and population genetics information by virulence gene testing and MLST.

Objective To study the genotypic and biological characteristics of catheter-related MRSE isolates and to further provide information for the diagnosis and prevention of catheter-related bloodstream infection. Methods Thirty strains of catheter-related MRSE isolates were collected from venosus blood and whole blood of 30 inpatients including 20 males and 10 females from Beijing Tongren Hospital, Capital Medical University from January 2006 to December 2009. The genetic features of these strains were determined by MLST. PCR was used to detect the icoA gene (encoding the polysaccharide intercellular adhesion associated with pathogenicity), and the antimicrobial susceptibility test was detected by disc diffusion test. Results A total of 15 STs were obtained from 30 strains ST259, ST20, ST2 and ST235 were common clones obtained from 17 strains. Four novel STs were found and uploaded to the MLST database (http://www. mlst. net), including ST259 (6 strains), ST260 (1 strain), ST261 (1 strain) and ST262 (1 strain). The ST259 was the dominant clone of catheter-related MRSE isolates in this hospital, and 3 strains carrying icaA gene were detected in this study. Conclusion Some ST259 isolates express high pathogenesis among the four novel STs, which may make them as the pandemic strains in nosocomial infection, and this would increase the difficulty of the prevention and control of nosocomial infection.