OBJECTIVE To realize AmpC genotypes of plasmid-mediated AmpC beta-lactamases in Escherichia coli and Klebsiella pneumoniae in Taizhou and Wenzhou,Zhejiang Province of China.METHODS Twenty strains of E.coli and 20 strains of K.pneumoniae which plasmid mediated AmpC beta-lactamases were detected by three methods such as three-dimensional-test of enzyme-extract,cefoxitin-triphase-test,cloxacillin-double-slips-synergia-test and detected ampC genotypes.RESULTS Five DHA genotypes,2 MIR and 2 FOX genotypes were found in 20 strains of E.coli with plasmid-mediated AmpC beta-lactamases.Seven DHA genotypes and 1 MIR genotype were found in 20 strains of K.pneumoniae with plasmid-mediated AmpC beta-lactamases.DHA and MIR genotypes were found in a strain of E.coli that ESBLs were positive but AmpC was suspicious.CONCLUSIONS The ampC genotypes of E.coli in Taizhou and Wenzhou are DHA genotypes mainly and then MIR and FOX genotypes;the ampC genotypes of K.pneumoniae in Taizhou and Wenzhou are DHA gene mainly and then MIR gene;their positive rates of DHA genes are 25%(E.coli) and 35%(K.pneumoniae),their positive rates of MIR genes are 10%(E.coli) and 5%(K.pneumoniae),and the positive rate of FOX is 10%(E.coli),no other genotypes were found.

OBJECTIVE To study antibiotic resistance and drug-resistant genes in AmpC and ESBLs-producing Klebsiella pneumoniae(KPN) in Wenzhou and Taizhou of Zhejiang Province.METHODS Antimicrobial susceptibilities of AmpC and ESBLs-producing KPN were tested by broth dilution motheds and VITEK 60 and VITEK 32;the genotypes of extended-spectrum beta-lactamases(ESBLs),cephalosporinase C(AmpC) and Genotypes AMEs were analyzed by polymerase chain reaction(PCR).RESULTS All tested KPN strains were susceptible to imipenem and resisitant to more other drugs.TEM genes were amplified in 20(100%) strains,CTX-MⅠgenes were examined in 18(90%) strains,SHV genes were examined in 2(10%) strains,DHA genes were tested in 17(85%) strains,MIR genes were examined in 3(15%) strains and 3 kinds of AMEs genes were tested in 18(90%) strains.CONCLUSIONS AmpC and ESBLs-producing KPN is multiple drug-resisitant,in which 2 to 6 resistant genes exist,Imipenem is the best drug.

Objectives To investigate the reliability of VITEK AMS for the detection of strains of E.coil and K.pneumoniae that produce ESBLs, and to determine the prevalence in our hospital of the two strains producing ESBLs.Method VITEK GNS 506 was used to detect ESBLs, and was compared with double disk synergy test. Results Among 48 strains of K.pneumoniase detected using VITEK GNS 506, 24 strains were found to be ESBLs positive, and 24 strains ESBLs negative. The result was consistent with that of double disk synergy test. Among 102 strains of E.coli detected using VITEK GNS 506, 41 strains were found to be ESBLs positive. The result was consistent with that of double disk synergy tests. 61 strains were found to be ESBLs negative detected using VITEK GNS 506. Among them, 19 strains were found to be ESBLs positive.Conclusion Detection ESBLs using VITEK AMS has some limitations in our area. Simple and reliable method should be used more with indicates for detection of ESBLs suitable for routine screening.

OBJECTIVE: To explore the inner quality of two kinds of single decoction solution. METHODS: The methods of HPLC, UV, TLC etc. were used for the qualitative and quantitative analysis. RESULTS: The contents of main components in single decoction solution of herbs in granule form were higher than those in slice form- CONCLUSION: The decocting rate of herbs in granule form is higher. It is worth popularizing.

We investigated molecular identification of a group of 14 strains of Aeromonas sp .,and genetic background of re‐sistance to beta‐lactams ,aminoglycosides .From January to December 2012 ,14 strains of Aeromonas sp .were collected from stool from diarrheal patients in enteric clinics in Ningbo First Hospital in Zhejiang Province ,China .Then ,molecular identifica‐tion by 16SrDNA ,23 kinds of beta‐lactamase genes ,6 kinds of aminoglycoside modifying enzyme genes ,6 kinds of 16srRNA methylase genes ,and 6 kinds of mobile genetic elements were analyzed by PCR .In addition ,genotyping and sample cluster a‐nalysis were performed .Results showed that 10 strains of A .hydrophila ,1 strain of A .aquariorum ,A .sobria ,A .entero‐pelogenes ,A .punctata were confirmed by 16SrDNA sequencing and arithmetic .Five kinds of beta‐lactamase genes ,4 kinds of aminoglycoside modifying enzyme genes ,and 3 kinds of mobile genetic elements were positive .BlaAQU of strain No .4(AQU‐2) and strain No .11(AQU‐3) were new subtypes .It’s suggested that identification of Aeromonas sp .should be performed by molecular identification method .This group of 14 strains of Aeromonas sp .conferred multidrug resistance .

Objective We conducted an in vivo study with the mouse model of Vib rio vulnificus infection to evaluate the efficacies of combination therapy with antimicrobia l agents. Methods Vibrio vulnificus (6.0?10 8 cfu/ml)was injected intraperitoneally into the right abdominal cavity. One hour, 2 hour and 3 hour after inoculation, 7 antimicrobial agents were given alone or in combination intraperitoneally at human therapeutic dose level. The numbers of survial mouse and the supermicrostr ucture change of organs w ere observed. 7 antimicrobial agents were Imipenem, Chloramphenicol, Doxycycline Hydrochloride, Netilmicin, Cefoperazone, Piperacillin, Levofloxacin. Res ults Two hours after infection, the mouse survival rates of groups trea ted with Chloramphenicol, Levofloxacin, Netilmicin, Cefoperazone was 100%. Howev er, the survival rates in the mouse treated by Piperacillin was 60% and the surv ival rate of in the mouse treated with Imipenem or Doxycycline Hydrochloride was 20%. The survival rate in the mouse treated with Cefoperazone combined with Lev ofloxacin, Cefoperazone combined with Netilmicin, or Netilmicin combined with Do xycycline Hydrochloride, was 100%. The supermicrostructure injure of the organs in the mouse recovered. Conclusions These results indicate that Chloramphenicol, Netilmicin, Cefoperazone and Levofl oxacin alone had satisfactory efficacy in the treatment of experimental Vibrio v ulnificus infection in mouse. The combination therapies of Cefoperazone with Lev ofloxacin, Cefoperazone with Netilmicin, and Netilmicin with Doxycycline Hydroch loride are more advantageous than using antimicrobial agent alone.

OBJECTIVE:To set up the quality standard for Xiaoyan San METHODS:Rheum officinale,Flos Lonicerae and Capejasmine were identified by TLC method The contents of emodin and chrysophanol in Xiaoyan San were determined by HPLC RESULTS:The linear range and average recovery of emodin was 21 12～105 6ng/ml(r=0 9 998,n=5) and 98 78%;The linear range and average recovery of chrysophanol were 29 28～146 4ng/ml(r=0 9 998,n=5) and 99 45% respectively CONCLUSION:The method is convenient,accurate and practicable It can be used for quality control in production of Xiaoyan San

Objective To evaluate the effect of dexmedetomidine on the expression of c-fos protein in the dorsal root ganglion neurons in a rat model of neuropathic pain (NP).Methods Seventy-two adult male SpragueDawley rats,weighing 180-240 g,were randomly divided into 3 groups (n =24 each) using a random number table:sham operation group (group S),group NP,and dexmedetomidine group (group Dex).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 350 mg/kg.The right sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 silk thread in NP and Dex groups.In group Dex,dexmedetomidine 50μg/kg was injected intraperitoneally once a day starting from the end of operation until the animals were sacrificed.The equal volume of normal saline was given instead of dexmedetomidine in S and NP groups.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before ligation (T0,baseline) and 3,7 and 14 days after ligation (T1-3).Eight animals were sacrificed after measurement of pain threshold at T13 and the dorsal root ganglions of the lumbar segments (L44) were removed for detection of c-fos expression (by immuno-histochemistry).Results Compared with group S,MWT was significantly decreased,TWL was shortened,and the expression of c-fos protein was up-regulated at T1-3 in NP and Dex groups.Compared with NP group,MWT was significantly increased,TWL was prolonged,and the expression of c-fos protein was down-regulated at T1-3 in Dex group.Conclusion Dexmedetomidine can inhibit upregulation of c-fos protein expression,thus attenuating NP in rats.

OBJECTIVE To investigate the changes in pathogenic bacteria and drug resistance in nosocomial(septicemia).METHODS The blood samples of inpatients were cultured with blood culture apparatus,VITEK-AMS were used to identify the pathogenic bacteria and conduct drug resistance test.RESULTS The proportion of(Gram-positive) cocci had been increasing,coagulase negative staphylococcus increased significantly,but the(proportion) of Staphylococcus aureus decreased significantly.The proportion of Gram-negative bacteria and fungi decreased too.Vancomycin and imipenem were the highest susceptible to Gram-positive and Gram-negative(bacteria),respectively.CONCLUSIONS Gram-negative bacteria are the major pathogens in nosocomial septicemia.But Gram-positive cocci had been increasing in the past years.Coagulase negative staphylococcus is the main pathogen in nosocomial septicemia.pathogenic bacteria are higher resistant to the commonly used antibiotics.

OBJECTIVE To investigate characters of molecular epidemiology of meticillin-resistant Staphylococcus aureus(MRSA) outbreak strains in an emergency intensive care unit(EICU),to follow-up the possible sources,understand transmission for infection,and determine preventive strategies.METHODS Pulsed-field gel electrophoresis(PFGE) was used to analyze the homology of MRSA strains,isolated from clinical patients′ infection sites and environment,and carried by patients and healthcare workers in EICU of our hospital in December,2004.RESULTS Six of 17 patients were infected by MRSA,and 7 strains were isolated((including) 2 strains from different sites of the same patient).Surveillance cultures of ward′s environments,(patients)′ nares and healthcare workers′ nares and hands were performed in the outbreak period.Five MRSA strains were isolated,including a strain from nares of a patient,a strain from a table-board of a procedure room,a strain from hand of a nurse,a strain from a bed bar,and a strain from ward′s air.PFGE typing of the 12 MRSA strains showed that all 7 strains isolated from patients′ infection sites and two strains from nares of a patient and hand of a nurse were of type A.Strains from a procedure room,bed bar and air were of types B,C and D,respectively.CONCLUSIONS MRSA′s source and its transmission route are elucidated by genotyping.MRSA appears to come from a patient′s nares and has been transferred in ward by hand of healthcare workers.