Objective To evaluate antibacterial activities of Cefoperazone-Sulbactam upon gram negative bacilli,and compare the differences in susceptibility between two different concentrations of Cefoperazone-Sulbactam combination disc.Method A total of 381 strains of commonly occurred gram negative bacilli were found from 2nd Affiliated Hospital of Zhejiang University School of Medicine,Zhejiang Provincial People's Hospital,The Third Hospital of Hangzhou and Hangzhou Hospital of Traditional Chinese Medicine respectively.Susceptibility test was conducted by K-B method using 75 μg/disc Cefoperazone plus with 75 μg/disc Sulbactam(150 disc)and 75 μg/disc Cefoperazone with 30 μg/disc Sulbactam(105 disc),respectively.Meanwhile the minimal inhibitory concentration(MIC)of Cefoperazone-Sulbactam was determined by standard agar dilution.The data were analyzed by using WHONET 5.4 and SPSS.Results Disc diffusion method was carried out to detect the antibacterial activities upon Escherichia coli,Klebsiella pneumonia,Pseudomonas aeruginosa and Acinetobacter baumannii,using 105 disc and 150 disc,respectively.The data indicated that consistency rates between these two different discs were 26.3%,79.2%,83.7%and 33%,respectively.The k-related sample test was performed by using SPSS version 10.0 and shown that the P value was less than 0.05.Upon those organisms mentioned above,the consistency rates between the antibacterial activities of 105 disc and those of agar dilution were 77.8%,89.6%,70.9%and 77%,respectively.When it was going to compare agar dilution vs.150 disc upon the susceptibility of those organisms the consistency rates were 27.3%,79.2%,61.6%and 30%,respectively.Compared with agar dilution,the error rates of those two different concentration discs revealed that the false susceptibility and false intermediate of 105 disc were higher than those of 105 disc.ConclusionsThe results of susceptibility test showed that 105 disc was more close to agar dilution than that of 150 disc.However,the 150 disc used in clinic led to increase in sensitivity of susceptibility test to organisms.

AIM: To study antipyretic, anti-inflammtory, analgesia and antisepsis effect of Renshenbaidu Pills.(Rhizouia Seu Radix Notopterygii, Radix Bupleuri, Radix Ginseup, etc) METHODS: The antipyretic effect of the pills was observed using rabbits fever model carrageenine caused foot swelling of rat, analgesic effect was tested by the method of writhes of mice, the method was applied antibacterial in vitro. RESULTS: Renshenbaidu Pills remarkably brought down the body temperature in experimental animal with fever 1.5 g/kg of Renshenbaidu Pills had the obvious anti-inflammatory effect and notable analgesic action on the reaction of writhes of mice induced by acestic acid, it had antisepsis effect. CONCLUSION: Renshenbaidu Pills are better than Renshenbaidu Powder, it has antiyretic, antiinflammatory, analgesia and antisepsis effects.

OBJECTIVE To detect AmpC ?-lactamases from isolates of extended spectrum ?-lactamases(ESBLs) positive Escherichia coli and Klebsiella pneumoniae,which were isolated from four hospitals in Hangzhou from 2005 to 2006,and to analyze the antimicrobial activity of clinically commonly used antibiotics in vitro.METHODS Amount of 324 ESBLs positive isolates including E.coli and K.pneumoniae were collected from Zhejiang Provincial People′s Hospital;1st Affiliated Hospital of Zhejiang University;2nd Affiliated Hospital of Zhejiang University and Hangzhou Traditional Chinese Medicine Hospital perspectivly.AmpC ?-lactamase was identified by disc screening testing and three dimensional test,and the genotypes of AmpC ?-lactamase were also determined by multiplex polymerase chain reaction(Multi-PCR).Minimal inhibitory concentrations(MICs) of ten clinically commonly used antibiotics were determined by agar dilution for AmpC ?-lactamases positive isolates,and the data were analyzed by WHONET 5.3.RESULTS AmpC ?-lactamase phenotype test revealed that 76 AmpC ?-lactamase positive isolates(23.5%) were identified among 324 ESBLs positive isolates of E.coli and K.pneumoniae from four hospitals in Huzhou.69.7% Of the AmpC ?-lactamase phenotype positive isolates were positively amplified by Multi-PCR.The value of MIC50 for carbapenem was lower 0.25 ?g/ml.We also found four carbapenem-resistant strains in this study.CONCLUSIONS We found that the incidence rate of AmpC ?-lactamase is high among the ESBLs producing E.coli and K.pneumoniae strains in Hangzhou.Carbapenem antibiotics have higher antimicrobial activity than other tested antibiotics.

Objective To evaluate the ability of matrix-assisted laser desorption/ionization time-offlight mass spectrometry (MALDI-TOF MS) in identifying species of Acinetobacter calcoaceticus-Acinetobacter baumannii complex,and investigate the species distribution of Acinetobacter calcoaceticus-Acinetobacter baumannii complex isolated in our hospital.Methods A total of 502 nonduplicate clinical isolates of Acinetobacter calcoaceticus-Acinetobacter baumannii complex were retrospectively collected from the second affiliated hospital of Zhejiang University between January 2012 and July 2012.All strains were re-identified by MALDI-TOF MS and were also verified by sequence analysis of 16S-23S rRNA gene spacer region.Results Among all the 502 strains of Acinetobacter calcoaceticus-Acinetobacter baumannii complex,identificaion results provided by MALDI-TOF MS were A.baumannii (431,85.9％),A.pittii (68,13.5％),A.calcoaceticus (3,0.6％).Sequence analysis of 16S-23S rRNA gene spacer region was used to identify all the 502 strains:403 (80.3％) were identified as A.baumannii,68 (13.5％) as A.pittii,28 (5.6％) as A.nosocomialis and 3 (0.6％) as A.calcoaceticus.MALDI-TOF MS correctly identified all the strains but erroneously identified all 28 strains of A.nosocomialis as A.baumannii,compared with sequence analysis of 16S-23S rRNA gene spacer region.Conclusions MALDI-TOF MS can be used as a fast,simple,reliable and excellently reproducible method to identify members of Acinetobacter calcoaceticus-Acinetobacter baumannii complex at low costs.MALDI-TOF MS is expected to be an ideal technique for routine clinical microbiology testing in the future.

Objective To investigate the prevalence and drug resistance of clinical Klebsiella vari-icola ( K. variicola ) isolates and to illuminate the mechanism of drug resistance in carbapenem-resistant strains. Methods Clinical K. variicola isolates were identified with matrix-assisted laser desorption/ioniza-tion time-of-flight mass spectrometry ( MALDI-TOF MS ) . The antimicrobial susceptibility profile of these strains was determined using broth microdilution. Resistance genes carried by carbapenem-resistant K. vari-icola strains were detected by PCR with specific primers. Multilocus sequence typing ( MLST) was used for molecular typing. A pan-drug resistant strain which was isolated from cerebrospinal fluid sample was ana-lyzed with whole genome sequencing ( WGS) . Results Twenty-six isolates were identified as K. variicola by MALDI-TOF MS. Results of the antimicrobial susceptibility test showed that there were 15. 4％ (4/26) re-sistant to carbapenem and 11. 5％ (3/26) unsusceptible to tigecycline. These strains were highly suscepti-ble to amikacin and gentamicin, which accounted for 96. 2％ (25/26). As for the third-and fourth-genera-tion cephalosporins, the resistance rate was 23. 1％ (6/26). All of the four carbapenem-resistant isolates carried the resistance genes of blaIMP-4 , qnrA/B and blaTEM , and one of them was also positive for blaNDM-1 gene. The fosfomycin resistance gene, fosA, was detected in three of them. Molecular typing analysis indica-ted these isolates belonged to two sequence types ( ST) of ST357 ( three strains) and ST1737 ( one strain) . Two plasmids were obtained from the pan-drug resistant strain by WGS, including IncFⅡ/FIB( k) type plas-mid (160 kb) that was highly homologous to LMG 23571 plasmid (GenBank: CP013986. 1) and IncHⅠ1B/FIB type plasmid (260 kb) sharing high homology with pIMP4 LL34 (GenBank: CP025964. 1). Be-sides the resistance genes mentioned above, the two plasmids also carried a variety of other genes that media-ted the resistance to aminoglycosides (strB, strA, armA, aac3-Ⅱd, aadA2), macrolides (msrE, mphE), chloramphenicol (catA2), sulfonamides (sulⅠ) tigecycline (tetA variant) and trimethoprim (dfrA16). However, no virulence genes were detected. Conclusions In general, the resistance profile of K. variicola was similar to that of Klebsiella pneumoniae, but the differences were that carbapenem-resistant K. variicola strains mainly belonged to ST357 and the leading causes of resistance were carrying the genes encoding IMP-4 and NDM-1 metalβ-lactamases. WGC analysis revealed that the pan-drug resistant K. variicola strain carried multiple drug resistance genes without virulence determinants, which might be resulted from the evo-lution of drug resistance.

In order to construct a recombinant replication deficient human type 5 adenovirus (Ad5) expressing a foot-and-mouth disease virus (FMDV) capsid protein, specific primers for P12A and 3B3C genes of FMDV-OZK93 were synthesized. The P12A and 3B3C genes were then amplified and connected by fusion PCR, and a recombinant shuttle plasmid pDC316-mCMV-EGFP-P12A3B3C expressing the FMDV-OZK93 capsid protein precursor P12A and 3B3C protease were obtained by inserting the P12A3B3C gene into the pDC316-mCMV-EGFP plasmid. The recombinant adenovirus rAdv-P12A3B3C-OZK93 was subsequently packaged, characterized and amplified using AdMax^TM adenovirus packaging system, and the expression was verified by infecting human embryonic kidney cell HEK-293. The humoral and cellular immunity levels of well-expressed and purified recombinant adenovirus immunized mice were evaluated. The results showed that rAdv-P12A3B3C-OZK93 could be stably passaged and the maximum virus titer reached 1×10^9.1 TCID₅₀/mL. Western blotting and indirect immunofluorescence showed that rAdv-P12A3B3C-OZK93 expressed the FMDV-specific proteins P12A and VP1 in HEK-293 cells. In addition, the PK cell infection experiment confirmed that rAdv-P12A3B3C-OZK93 could infect porcine cells, which is essential for vaccination in pigs. Comparing with the inactivated vaccine group, the recombinant adenovirus could induce higher FMDV-specific IgG antibodies, γ-IFN and IL-10. This indicates that the recombinant adenovirus has good immunity for animal, which is very important for the subsequent development of foot-and-mouth disease vaccine.
Adenoviridae/genetics* ; Adenoviruses, Human/genetics* ; Animals ; Antibodies, Viral ; Capsid/metabolism* ; Capsid Proteins ; Foot-and-Mouth Disease/prevention & control* ; Foot-and-Mouth Disease Virus/genetics* ; HEK293 Cells ; Humans ; Mice ; Recombinant Proteins/genetics* ; Serogroup ; Swine ; Viral Proteins ; Viral Vaccines/genetics*