Objective To investigate the effect of salinity and temperature on motility of Vibrio parahaemolyticus. Methods V.parahaemolyticus was inoculated on swarming or swimming agar plates containing different amounts of salinity (0.5%, 1.0%, 2.0%, and 4.0% NaCl, respectively), followed by incubation at 26 or 37℃, before the diameters of bacterial lawns were measured .Results and Conclusion The swarming motility was not affected by salinity , while the swimming motility was positively correlated with salinity .Maximum swimming occurred in 2.0% NaCl, and displayed a slight decline in salinity of 4.0%.Both swimming and swarming were affected by temperature , and the motility was signifi-cantly enhanced in 37℃vs 26℃.These results indicate that both salinity and temperature can modulate the motility of V. parahaemolyticus.

Objective To study the influence of Noradrenergic systems in Lateral Hypothalamus Area (LHA) on small intestine moving. Methods The effect of noradrenalin、noradrenalin+phentolamine、noradrenalin+propolol、phentolamine on small intestines electro-activity of rats was detected by external alimentary canal electrodes and central nervous system stereo-configuration technology. Results After injecting 2 g/L noradrenaline (NE) the cyclic period of MMC extended, the ratio of the active time to the cyclic period and the number of the fast wave within the active time per minute reduced. NE inhibited the electro-activity of small intestine of rats, and the average effective time was (36.86?7.39)min. Hence injecting 5 g/L PE alone to LHA raised the ratio of the MMC active time to the cyclic period. Phentolamine presented a slight excitability on the electro-activity of small intestine of rats. Conclusion The NE in LHA showed the inhibitive myoelectric activity on small intestines, and this effect was introduced through a acceptor,The reacceptor in LHA may function in the inhibition of the electro-activity in small intestines.

Objective To study the impact of QseBC on the motility of Salmonella enterica serovar Typhi ( S.Typhi ) . Methods The motility of wild-type ( WT) and null mutants (ΔqseB and ΔqseC) at mid-log phase was investigated by swimming assay.Quantitative RT-PCR was carried out to calculate the transcriptional variation of flhD and qseB among WT,ΔqseB andΔqseC.QseB overexpressing strain was constructed to compare its motility and flhD expression with the wild-type control.Results The result of motility assay showed that the motility of ΔqseB was similar to that of the WT strain , while the motility of ΔqseC was much lower than that of WT .qRT-PCR revealed that compared with WT , the expression of flhD was significantly decreased in ΔqseC while the expression of qseB was increased considerably .The motility of QseB overex-pressing strain was lower .Conclusion The expression of flhD may be regulated by QseBC which has an effect on the motil-ity of S.typhi, and the overexpression of QseB may inhibit the motility .

Objective To investigate the transcriptional autoregulation of PhoP/PhoQ under different growth conditions in Yersinia pestis.Methods The entire promoter region of YPO1635 was amplified and cloned into the pRW50 vector containing a promoterless lacZ reporter gene.The recombinant LacZ reporter plasmid was transformed into the wild-type strain (WT) and the phoP mutant strain (ΔphoP),respectively,to measure the promoter activity (the β-galactosidase activity) of the target gene in WT and ΔphoP by using the β-galactosidase enzyme assay system.Total RNAs were extracted from WT and ΔphoP strains,and primer extension assay was employed to detect the promoter activity by examining the amount of primer extension products of YPO1635 in WT and ΔphoP.Results The LacZ fusion results showed that the transcription of YPO1635 was positively regulated by PhoP under L-TMH and brain-heart infusion(BHI) conditions,but it was not regulated in H-TMH medium.The primer extension assay detected two transcriptional start sites located at 90 and 118 bp upstream of the translation initiation site of phoP,named P1 and P2,respectively.Under low Mg2+ TMH conditions,the promoter activity of P1 rather than P2 was positively regulated by PhoP.Under high Mg2+ TMH conditions,the promoter activities of both P1 and P2 showed no obvious difference in the WT and ΔphoP strains.Under rich BHI conditions,both promoters were under negative control of PhoP.Conclusion Different autoregualtion patterns of PhoP/PhoQ under different growth conditions would help Y.pestis to quickly adapt to the changing living environment.

OBJECTIVETo establish HPLC fingerprint for identification and evaluation of the quality of Trachelospermum jasminoides.

METHODThe analysis was performed on a ZORBAX Eclipse XDB-C18 analytical column (4.6 mm x 150 mm, 5 microm) with H2O (A) and methanol (B) as mobile phases in gradient mode. The elutin conditions were 0-15 min, changed from 10% B to 30% B; 15-40 min, to 40% B; 40-60 min, to 60% B. The column temperature was set at 30 degrees C and the flow rate was 0.8 mL x min(-1) with the detection wavelength of 230 nm.

RESULTThe HPLC chromatographic fingerprint of T. jasminoides, showing 19 characteristic peaks, was established from 14 lots of T. jasminoides. The similarity of every lot of T. jasminoides was calculated as good in general, the similarity of 10 lots was above 0.9 and the other 4 lots had low similarity.

CONCLUSIONThe chromatographic fingerprint of T. jasminoides with high characteristics and specificity can be used to control its quality.

Objective To study the transcriptional regulation of vp1667 by H-NS in Vibrio parahaemolyticus.Methods Total RNAs were extracted from Δhns and WT strains.Quantitative RT-PCR was carried out to calculate the transcriptional variation of vp1667 between Δhns and WT.Primer extension assay was also employed to detect the transcription start site and the promoter activity (i.e.the amount of primer extension products) of vp1667 in Δhns and that in WT.The promoter DNA region of vp1667 was amplified, purified, and cloned into the corresponding restriction endonuclease sites of pHRP309 that harbors a gentamicin resistance gene and a promoterless lacZ reporter gene.The recombinant pHRP309 plasmid was transformed into Δhns and WT, respectively, while β-galactosidase activity in cellular extracts was measured using a β-galactosidase enzyme assay system.The over-expressed His-H-NS was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns.The electrophoretic mobility shift assay (EMSA) and DNaseⅠ footprinting were then applied to analyze the DNA-binding activity of His-H-NS to vp1667 promoter region in vitro.Results and Conclusion The primer extension assay detected one transcription start site for vp1667, which was located at 28 bp upstream of vp1667, and its transcribed activity was under the negative control of the H-NS.The EMSA and DNaseⅠ footprinting assay results showed that His-H-NS was unable to bind to the promoter-proximal DNA region of vp1667, suggesting that H-NS indirectly inhibits the transcription of vp1667.

OBJECTIVE To investigate the produce of extended-spectrum ?-lactamases(ESBLs) and the presence of genotype of the ?-lactamases-encoding genes in Klebsiella pneumoniae isolated from the 98th Hospital of PLA,Huzhou,Zhejiang Province,China.METHODS Twenty-five strains of K.pneumoniae were isolated from the inpatients between Sep 2005 and Apr 2006.ESBLs were tested by phenotypic confirmatory tests recommended by CLSI.Twenty-one kinds of ?-lactamases genes of blaTEM,blaSHV,blaLEN,blaOKP,blaCTX-M-1 group,blaCTX-M-2 group,blaCTX-M-9 group,blaOXA-1 group,blaOXA-2 group,blaOXA-10 group,blCARB,blaPER,blaVEB,blaGES,blaLAP,blaDHA,blaACT/MIR,blaCMY/MOX,blaFOX,blaCMY/LAT,and blaACC were analyzed by PCR and verified by DNA sequencing.RESULTS In 25 strains of K.pneumoniae,the positive,negative,and "uncertainty" rates of ESBLs were 56.0%,20.0%,and 24.0%,respectively.The positive rate of genes of blaTEM,blaSHV,blaCTX-M-1 group,blaOXA-10 group,blaLAP,and blaDHA were 80.0%,4.0%,4.0%,80.0%,4.0% and 32.0%,respectively.The 15 kinds of rest genes were all tested negative.The total positive rate of 21 kinds of ?-lactamases gene was 92.0%.Among them,the blaLAP-2 gene sequence of the HZ12593 strain has been registered in GenBank(GenBank Accession Number: EU529981).CONCLUSIONS There are higher rate of ESBLs-producing strains in K.pneumoniae isolated from the inpatients,and at least 6 kinds of ?-lactamases gene existed.Both genes of blaTEM and blaOXA-10 group are the most common genotypes.Carring blaDHA Gene may influence the result of phenotypic confirmatory test for ESBLs in K.pneumoniae.

Objective To optimize the preparation of nanoparticles encapsulating antisense oligodeoxynucleotides in a-butyleyanoacrylate carrier (ASODN in NP) and investigate their stability. Methods ASODN in NP were prepared by interfacial polymerization of butyleyanoacrylate (BCA). The formulation and technology of the prepared NP was optimized by using orthogonal design based on the single-factor experiment. The morphology of NP was examined by transmission electron microscope; The size and size distribution of NP were determined by Malvern laser granularity equipment;The encapsulation efficiency and drug loading were determined by HPLC; The ability of protecting oligodeoxynucleotides from serum was investigated on a 20% polyacrylamide-7 Murea sequencing gel (PAGE). Results The nanoparticles in the optimal conditions were of regular spherical surface and discrete. The average size was 97.1 nm,the average encapsulation efficiency and drug loading of ASODN in NP were 96.7% and 10.1% respectively; The oligonucleotides were more efficiently protected from degradation by nucleases than by oligonucleotides adsorbed into nanospheres.Conclusion ASODN in NP has good stability,encapsulation efficiency,drug loading and great potential for ASODN delivery.

Animals ; Bacterial Proteins ; genetics ; metabolism ; Biofilms ; growth & development ; Gene Expression Regulation, Bacterial ; Lipopolysaccharides ; chemistry ; metabolism ; Operon ; Promoter Regions, Genetic ; Protein Binding ; Siphonaptera ; microbiology ; Species Specificity ; Transcription, Genetic ; Transferases ; genetics ; metabolism ; Virulence ; Yersinia pestis ; genetics ; metabolism ; pathogenicity ; Yersinia pseudotuberculosis ; genetics ; metabolism

Objective To explore the correlations of intracranial pressure (ICP) with changes of neuron specific enolase (NSE),D-Dimer (D-D) and C-reactive protein (CRP) levels in patients with severe traumatic brain injury.Methods A serial of 35 patients with severe traumatic brain injury,admitted to our hospital from January 2012 to January 2014,were chosen as experimental group,and 20 healthy subjects performed physical examination in our Physical Examination Center at the same period were as controls.ICP monitoring was performed in these 35 patients.The patents were divided into two groups according to ICP:severely elevated ICP group (＞40 mmHg) and moderately elevated ICP group (20-40 mmHg).The NSE,D-D and CRP levels were measured,and these data were compared with those from the control group.The correlations of ICP with changes of NSE,D-D and CRP levels were analyzed.Results The levels of NSE,D-D and CRP in the severely elevated ICP group and moderately elevated ICP group were obviously higher than those in the control group ([12.11 ±2.35] lg/L,[0.39±0.61] mg/L,[3.72±0.69] mg/L) (P＜0.05).The levels ofNSE,D-D and CRP in the severely elevated ICP group ([104.08±7.90] μg/L,[1.55±0.26] mg/L,[47.66±8.60] mg/L) were also obviously higher than those in the moderately elevated ICP group ([61.89±30.35] μg/L,[0.93±0.32] mg/L,[30.87±9.84] mg/L)(P＜0.05).Significant positive correlations were noted between ICP and changes ofNSE,D-D and CRP levels in the patient group (regression equation:ICP=18.598+0.256 NSE [t=7.200,P=0.000],ICP=10.779+23.955D-D [t=10.29,P=0.000],ICP=9.932+0.771 CRP [t=8.423,P=0.000]).Multivariant stepwise regression analysis indicated the closest correlation between ICP and D-D (multiple correlation coefficient=0.873,coefficient of determination=0.762,F=105.917,P=0.000).Conclusions Significant positive correlations can be noted between ICP and changes of NSE,D-D and CRP levels,and the closest correlation is between ICP and D-D in patients with severe traumatic brain injury.The combined application of ICP and NSE,D-D and CRP levels can promote the diagnosis and treatment of severe traumatic brain injury patients.