Objective To investigate the infection status and drug resistance of imipenem‐resistant Pseudomonas aeruginosa (PAE) isolated from the submitted specimens from children patients in our hospital between October 2012 to September 2013 for mastering the antimicrobial resistance status of Pseudomonas aeruginosa infection among children in Guangzhou and the occurrence situation of imipenem resistant strains .Methods The detection situation of Pseudomonas aeruginosa from infected children during this period was analyzed and the VITEK 2 Compact analyzer produced by the France bioMerieux company was used to identify the bacteria .The minimal inhibitory concentration(MIC) of imipenem resistant Pseudomonas aeruginosa to 11 kinds of antibiotics was detected .Results 161 strains of Pseudomonas aeruginosa were detected from 36 600 specimens ,including 24 strains(14 .9% ) of imipenem resistant Pseudomonas aeruginosa ,the positive strains were mainly originated from phlegmy (50 .3% ) ,the isolation was highest in ICU (27 .4% ) and NICU(21 .8% ) .The drug resistance rate of imipenem‐resistant P .aeruginosa was higher to ceftriax‐one sodium(91 .7% ) ,cefoperazone(29 .0% ) ,ceftazidime(29 .0% ) ,cefoperazone /shubatan(29 .0% ) and aztreonam (25 .0% ) .Con‐clusion Imipenem‐resistant Pseudomonas aeruginosa has the higher detection rate among children in this area and usually has re‐sistance to multiple antibiotics ,which should be paid more attention to and antibiotics should be rationally used .

Objective To investigate the most suitable anaesthesia method for the tension-free herniorrhaphy.Methods A total of 422 unilateral inguinal hernia cases from 2002 to 2005 were collected and randomly divided into the local anaesthesia group and epidural anaesthesia group. Observation indices and some relative data, such as operative duration, date of ambulation, date of foodintake, length of hospital stay, operation-correlated complications, anaesthesia complications, usage rate of ancillary drug, satisfactory rate for anesthesia, cost of hospitalization, were included and recorded in the questionnaire, and all the patients who took the tension-free herniorrhaphy were asked to answer it as the follow-up research. Results It was found that the occurrence of postoperative anaesthetic complications, the cost of hospitalization, length of stay of local anaesthesia group were significantly less than those of epidural anaesthesia group, and the date of moving and the date of foodintake were also significantly earlier than those of the other group (P0.05). Conclusion The local anaesthesia is suitable for most of the tension-free herniorrhaphy, and it may be used as the conventional anaesthetic method.

Objective To study the synergistic antitumor effects of quercetin and cisplatin in human prostate cancer PC3 cells and LNCaP cells.Methods Twelve h after PC3 cells or LNCaP cells were seeded,different dose of quercetin (10 μmol/L,20 μmol/L,40 μmol/L,80 μmol/L,160 μmol/L) or cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L),or quercetin (20 μmol/L) + cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L) were added for48 h and then the antiproliferative effects were detected with MTT assay.After incubated with quercetin (20 μmol/L) or cisplatin (0.05 μ mol/L),or quercetin (20 μ mol/L) + cisplatin (0.05 μmol/L) for48 h,cell cycle distribution and apoptosis of PC3 cells or LNCaP cells were detected by flow cytometer,PI and Annexin V staining.Protein expression was detected by Western blotting.Results After treatment with quercetin or cisplatin alone,the IC50 were (0.99 ± 0.13) μmol/L,(0.75 ± 0.09) μmol/L and (91.60 ± 6.10) μ mol/L,(72.90±4.70) μ mol/L for LNCaP cells or PC3 cells,respectively;The IC50 were (0.11±0.06)μ mol/L,(0.07±0.02) μmol/L for quercetin + cisplatin treatment (Compared with quercetin,P＜0.01 ;Compared with cisplatin,P＜0.05.After treatment with cisplatin or quercetin + cisplatin for 48 h,the S phase percent of LNCaP cells or PC3 cells were (22.4±2.7)％,(31.2±2.4)％ and (20.1±1.6)％,(31.0±2.5)％,respectively,(Compared with control,P＜0.05,however,treatment with quercetin alone has no significant difference (Compared with control,P＞0.05).After treatment with cisplatin or quercetin + cisplatin for 48 h,the apoptotic percent of LNCaP cells or PC3 cells were (14.8 ± 1.9) ％,(39.6 ± 3.1) ％ and (11.5± 1.2) ％,(34.1 ±3.3) ％,respectively,(compared with control,P ＜ 0.05,however,treatment with quercetin alone had no significant difference (compared with control,P＞0.05).After treatment with quercetin alone for 48 h,the activation of caspase-3,caspase-8 and caspase-9 were slightly increased,the expressions of Bax and p21 were up-regulated,the expressions of Bcl-2 and CDK2 were down-regulated.Furthermore,these effect of cisplatin and quercetin + cisplatin were significantly enhanced (compared with quercetin,P＜0.05;compared with quercetin,P＜0.01,respectively.Conclusions The combination modality with quercetin and cisplatin has a better treatment effect in vitro not only in androgen-dependent LNCaP cells but also in androgen-independent PC3 cells.

Objective To understand and analysis of the pathogens distribution and drug resistance of nosocomial infection in children′s hospital,so as to provide reliable scientific basis for the prevention and control of hospital infection.Methods 396 cases of upper respiratory tract specimens were collected from pediatric patients with nosocomial infection.These specimens were detected by sputum specimens conventional methods of microorganism cultivation,and K-B method was used to determine the bacteria sensi-tivities to clinical common drug.Results There were 225 cases of specimens were pathogen positive among all the 396 specimens, and 234 strains of bacteria were isolated in all.The positive isolated rate was 56.8%(225/396).Among the 234 isolated strains, Gram negative bacteria accounted for 72.6%(170/234),and Klebsiella occupied the first place[49.4%(84/170)].Gram positive bacteria accounted for 23.5%(55/234),and Staphylococcus had the highest isolated rate in Gram positive bacteria[58.2% (32/55)].In all the 9 kinds of clinical common antimicrobial agents,imipenem had high drug sensitivity to the 234 isolated strains,and the aminoglycosides came next.Conclusion It is necessary for the pediatric patients with nosocomial infection to collect upper re-spiratory tract specimens for bacteriologic studies and drug sensitivity tests.

Objective To evaluate the clinical significance of telomerase in the diagnosis of lung cancerusing SROC curve method.Methods Looking“telomerase”and“lung cancer”as keywords,retrieving journalspublished within past 20 years in order to incorporate into literatures and to collect data .To conduct the SROC analysisusing Meta -DiSc 1.4 software.Results 1.Twenty -two documents were sampled as tissue specimensand the heterogeneity was relatively large (P =0.017).To analyze the data with random effective models ,thecombined sensitivity and specificity were 0.788(0.761 -0.814)and 0.955(0.936 -0.969),respectively.TheSROC AUC area under the curve was 0.9515,SE(AUC) =0.0145.2.There was no heterogeneity(P =0.633)amongthe 10 lavaged literatures.By use of fixed effects model for data analysis ,the combined sensitivity and specificitywere 0.777(0.734 -0.816) and 0.922(0.888 -0.948),and SROC AUC area under the curve was0.9369,SE(AUC) =0.0141.Conclusion Telomerase is a ideal tumor marker,and the detection of telomeraseactivity in lavage fluid is stable and accurate in clinical diagnosis .

Objective To investigate the changes of pathogens distribution and antimicrobial resistance in children with urinary tract infection (UTI) in 10 years. Methods The results of urine culture and drug sensitivity in children with UTI from January 2001 to December 2003, and from January 2011 to December 2013 were retrospectively analyzed.Results In recent 10 years, there was no obvious change in the ratio of gram-negative bacteria to gram-positive bacteria. Escherichia coli was still the main bacteria causing UTI in children. The detection rate of enterococcus was signiifcantly increased from 18.3%in 2011-2013 to 7.5%in 2001-2003 (P<0.05) and it had become the second pathogenic bacteria. The isolation rate of ESBLs producing strains was signiifcantly higher in 2011-2013 than in 2001-2003 (P<0.05). The rate of Escherichia coli sensitive to imipenem re-mained at 100%and it is also sensitive to enzyme inhibitors antibiotics and nitrofuranto. Sensitivities to antibiotics were changed in different species of enterococcus. Conclusions The distribution of pathogens and antimicrobial resistance in children with UTI are constantly changing. The clinician should pay close attention to changes of epidemiology in the region and hospital and rational use of antimicrobial drugs.

Objective To explore the main pathogenic bacteria and antibiotic resistance patterns in children with bacterial diar‐rhea from Guangzhou region .Methods Regular bacterial culture of stool samples from children with suspicious bacterial diarrhea was performed to isolate the pathogen during 2011 to 2012 ,followed by the analysis of its composition and serum type ,ward distri‐bution characteristics and drug resistance to 12 antimicrobacterial drugs .Results 416 strains of pathogenic bacteria were isolated from diarrhea children during 2011-2012 ,in which salmonella ,enteropathogenic E .coli ,Campylobacter jejuni and Candida albicans isolates accounted for 53 .61% ,37 .98% ,5 .29% and 1 .68% respectively .Drug resistance rate of the main strains to 12 antimicrobi‐al agents was 85 .25% to ampicillin ,54 .28% to compound sulfamethoxazole ,44 .70% to cefotaxime ,42 .53% to ceftriaxone , 40 .66% to chloramphenicol ,23 .55% to ceftazidime ,23 .36% to aztreonam ,14 .88% to ciprofloxacin ,8 .07% to cefepime ,7 .99% to cefperazone/sulbactam ,7 .42% to piperacillin/tazobactam respectively ,and no resistance to imipenem was detected .Conclusion The pathogenic bacteria causing diarrhea mainly includes salmonella ,pathogenic e .coli ,campylobacter jejuni in children from guang‐zhou region ,the top five sensitive antimicrobial reagents for the main strains includes imipenem ,piperacillin/tazobactam ,cefpera‐zone/sulbactam ,cefepime and ciprofloxacin .

Objective To observe the clinical effect of compound remifentanil etomidate on gastrointestinalendoscopy and treat-ment among aged patients .Methods Divided 400 aged patients who got painless gastrointestinalendoscopy and treatment into two groups ,where one was observation group(200 aged patients) that got compound remifentanil etomidate ,while the other was control group which got fentanyl combined propofol .Compared and observed two groups on oxyhemoglobin saturation ,heart rate ,systolic-pressure ,diastolic pressure ,incidence of adverse reaction ,recovery time ,and time of leaving operating room .Results There was no obvious difference between observation group and control group on oxyhemoglobin saturation ,heart rate ,systolicpressure ,diastolic pressure before examination(P>0 .05);while there was distinct difference between examining and reviving ,which showed statisti-cal significance(P<0 .05) .The incidences of bucking ,dysphoria ,respiratory depression ,nausea and vomiting ,and dizziness in the two groups respectively were 3% 、2% 、5% 、3% 、11% and 8% 、11% 、13% 、8% 、27% ,which indicated the incidence of adverse reac-tion in observation group was obviously lower than that of control group ,where there was statistical significance (P<0 .05) .The time of recovery and the time of leaving operating room in two groups respectively were (3 .5 ± 1 .3)min、(9 .5 ± 1 .5)min and (7 .5 ± 3 .4)min、(18 .5 ± 4 .6)min ,which showed the time of observation group was lesser than that of control group ,where there was sta-tistical significance(P<0 .05) .Conclusion During the gastrointestinalendoscopy to aged patients ,compound fentanyl etomidate was safety ,less adverse reaction ,efficiency ,and strong controllability .

Objective To clone and express Staphylococcus aureus drug resistance adenylyltransferase gene in E .coli BL21 ,and to make the foundation for its function research .Methods Primers were designed on the basis of adenylyltransferase gene in gen‐bank ,PCR was used to amplify adenylyltransferase gene using Staphylococcus aureus genomic DNA as template .The obtained PCR production was attatched with pGEX‐4t‐1(+ ) plasmid ,and transformed into E .coli BL21 (DE3) .The recombinant plasmid was di‐gested by double enzyme digestion and identified by gene sequence .The recombinant protein was induced to expression by IPTG and identified by Western‐blotting .Results Using Staphylococcus aureus genome as a template ,the target fragment about 800 bp was successful amplified .After enzyme‐cutting and DNA‐sequencing ,the target fragment showed that the ORF begin with ATG ,end with TAG ,783 bp in length ,the predicted isoelectric point and molecular weight were 7 .75 and 29 × 103 ,and it was homology 99%homology with the reported sequence gene in genbank .SDS‐PAGE and Western‐blot showed the molecular weight of recombinant fusion protein was about 55 × 103 .Conclusion Adenylyltransferase gene of Staphylococcus aureus was successfully cloned and ex‐pressed in E .coli as a fusion protein ,which makes the foundation for the research of its function .

Objective To perform the amplification ,sequencing and prokaryotic expression of APH (3′′)‐Ⅰ and AAC (2′)‐Ⅰgenes from the clinically isolated gzch810 strain(SM gzch810)of Stenotrophomonas maltophilia to provide the basic materials for the next step functional test .Methods The SM gzch810 genome chromosome was extracted ,the APH (3′′)‐Ⅰ ,AAC (2′)‐Ⅰ whole genes were amplified by PCR and sequenced after being cloned into pMD18‐T vector .The recombination were subcloned into pGEX‐4T‐1 vector and the expression of the recombinant APH (3′′)‐Ⅰ and AAC (2′)‐Ⅰ were analyzed by SDS‐PAGE .Results The 800bp and 550bp DNA fragments of APH(3′′)‐Ⅰ ,AAC(2′)‐Ⅰ gene were amplified from SM gzch810 by PCR and sequenced ;the sequence comparison analysis showed that DNA and amino acid sequence identities of APH (3′′)‐Ⅰand AAC (2′)‐Ⅰ genes with other strains were 91% and 95% respectively .The sequence of APH (3′′)‐Ⅰand AAC(2′)‐Ⅰ of SM gzch810 were submitted to GenBank(accession number :HQ315852 and HQ315853);two major protein bands corresponding to the expected recombinant GST‐TP fusion proteins (56 × 103 and 46 × 103 respectively) were identified by SDS‐PAGE .Conclusion APH(3′′)‐Ⅰand AAC(2′)‐Ⅰgene of SM gzch810 are successfully cloned and expressed ,which lays a good foundation for further detecting corresponding antibi‐otic resistance and functional evaluation of above two kinds of recombinant E .coli .