Objective To investigate the expression and significance of E-selectin in lung tissue of rat of acute necrotizing pancreatitis (ANP)-induced lung injury.Methods Rats were randomly divided into control and ANP groups.ANP model was induced by retrograde injection of sodium taurocholate into the biliary and pancreatic duct.The rats were sacrificed at 3,6,12 and 24 h.Serum amylase,albumin,Ca2+ and lung wet/dry (W/D) weight ratio were determined,and pancreas and lung tissues underwent routine pathological examination and histopathological scores were evaluated.The expression of E-selectin mRNA and protein in lung tissue of rat was measured by real-time fluorescence quantitative RT-PCR and immunohistochemistry.Results In ANP group at 6h,the serum level of amylase,histopathological scores of pancreas and lung tissues,lung W/D weight ratio were (3978 ± 476) U/L,8.22 ± 0.63,( 12.31 ± 1.58,5.21 ± 0.20,which were significantly higher than those in control group [ (843 ± 90)U/L,0.22 ± 0.36,0.13 ± 0.34,4.46 ±0.17,P ＜ 0.05 or ＜ 0.01 ],the serum level of albumin was (12.9 ± 1.0)g/L,which was significantly lower than that in control group [ ( 15.6 ± 0.7 ) g/L,P ＜ 0.01 ],the serum level of Ca2 + at 12 h was ( 2.33 ±0.15) mmoL/L,which was significantly lower than that in control group [(2.57 ± 0.23)mmoL/L,P ＜ 0.01 ].E-selectin was located in the cell membrane and cytoplasm of vascular endothelial cells.Expression of E-seleetin mRuNA and protein in the lung tissue at 6h was significantly higher than those in control group (3.51 ±0.45 vs 0.95 ± 0.16,0.174 ± 0.019 vs 0.060 ± 006,P ＜ 0.01 ).Conclusions Pulmonary expression of E-selectin of vascular endothelial cell is significantly up-regulated in a time dependant manner when ANPinduced ALI occurs; and it is involved in leukocyte mural and extravasation,and can promote lung injury.

Objective To explore the role of IL-1β in capillary leak syndrome by observing the alterations of AQP-1 expression,apoptosis,and ultrastructural of vascular endothelial cells under the action of IL-1β.Methods Umbilical vein endothelial cells (UVEC) in vitro were randomly allocated into 3 groups:time,concentration,and control.In the time group,UVECs were treated with culture medium containing 20 μg/L IL-1β for3 h(T1),8 h(T2),12 h(T3) and 24 h(T4).In the concentration group,UVECs were treated with culture medium containing 0.2 μg/L(C1),2 μg/L(C2) and 20 μg/L(C3) IL-1β for 24 h.In the control group,UVECs were treated with culture medium without IL-1β for 24 h.The changes of AQP-1 mRNA and protein expression were detected by real-time PCR and Western blot.Apoptosis was detected by flow cytometry,and cell ultrastructural changes were observed by electron microscopy.Results AQP-1 mRNA and protein expression of T1-T4 in the time group and C1-C3 in the concentration group were lower than those of the control group (P ＜ 0.05).The apoptotic rate was increased,and mitochondrial swelling,vacuolar degeneration,karyolysis and necrosis were observed under electron microscopy.These were more pronounced with time or concentration increases.Conclusions IL-1β can cause a decrease of AQP-1 mRNA and protein expression,increase in apoptotic rate and increase in damage to the cells'ultrastructure.This is an important reason for damage to the vascular endothelial barrier and may be associated with capillary leak syndrome.

Objective To observe the effect of the different bore diameter membranes on technological parameters of Radix Polygoni Multiflori Granula and to optimize the process of membrane filtration.Methods Three different membranes were tested under different parameters of operation to observe the changes in membrane flux and the content of effective components by Radix Polygoni Multiflori Granula.Results The membrane(50 nm)had the great flux,the transfering rate of stilbene glucoside was the highest;The optimum conditions:the operation differential pressure was 0.8-1.2 MPa;The operation temperature was 50-60 ℃;the membrane surface flow rate was 3.0 m/s.When the volume of filtration solution was condensed to 1/10 of former,entering the same volume deionized water into it,and when the effective components transfering rate reached 80%,the filtration was finished.Using the strong acid and strong alkali to wash it in turn,the flux could revive above 90%.Conclusion It can be obtained a good result that adopts the technology of ceramic membranes microfiltration to refine Radix Polygoni Multiflori Granula.This can provide the foundation for the application of ceramic membrane microfiltration in the refinement of water extraction of other Chinese materia medica.

Objective To investigate the mechanism of cyclic adenosine monophosphate / protein kinase A signal transduction pathway in severe acute pancreatitis(SAP)-associated lung injury.Methods Seventy-two male healthy SD rats were completely randomized into three groups:sham operation (SO) group(n =8),SAP group and SAP plus H89 (cAMP inhibitor) group,then the latter two groups were divided into four sub-groups with eight rats in each sub-group according to the sampling time of 3,6,12 and 24 h,and the total numbers of groups were nine.The content change of TNF-α and IL-1β in serum,protein levels of cAMP-dependent protein kinase catalytic subunit (PKA C) and phosphorylated vasodilator-stimulated phosphoprotein(p-VASP) and the expression of VSAP mRNA in lung tissue were detected by enzyme-linked immunosorbent assay (ELISA),immunohistochemistry and quantitative real time PCR,respectively.Pathological changes of the pancreas and lung tissues were also observed.Results Compared with the SO group,the serum levels of TNF-α and IL-1β in the SAP group were obviously increased at different time points(P ＜0.05).Pathological changes of the pancreas and lung tissues were aggravated significantly.The protein levels of PKA C,p-VASP and the expression of VSAP mRNA in lung tissue were increased significantly (P ＜0.05)which peaked at 12 h in the SAP group [TNF-α was (266.07 ± 17.14) pg/mL,IL-1β(169.17 ±25.92) pg/mL,PKA C(210.69 ±6.32) × 103,p-VASP (56.62 ±0.57) × 103,VASP mRNA(2.06 ±0.21)],which had positive correlation with the serum level of TNF-α and IL-1β.Compared with the SAP group,pathological changes of the pancreas and lung tissues were alleviated significantly,the protein levels of PKA C,p-VASP and the expression of VSAP mRNA in lung tissue were decreased significantly in the SAP plus H89 group at different time points(P ＜ 0.05).Conclusion The cyclic adenosine monophosphate / protein kinase A signal transduction pathway is found to participate in the pathological process of SAP-associated lung injury through the up-regulations of TNF-α,IL-1 β and phospho-VASP.

OBJECTIVE: To establish a RP-HPLC method for the determination of the content of hesperidin in Yangweishu tablets.METHODS: The samples were separated on octadecyl silane bonded gel silica column(150 mm?4.6 mm,5 ?m) with the mobile phase consisted of methanol-7% acetic acid solution(32∶68) under a detection wavelength of 283nm.RESULTS: The linear range of hesperidin was 49.6～892.8 ng(r=0.999 9) with an average recovery rate of 99.02%(RSD=0.45%,n=9).CONCLUSION:The method is simple,accurate,reproducible and applicable for the quality control of Yangweishu tablets.

Objective To explore the carry rate and antimicrobial resistance of Staphylococcus aureus (S.aureus) among primary school students.Methods Nasal swab samples were collected from healthy primary school students in Guangzhou.Antibiotic susceptibility testing was applied to test S.aureus strains.Results The overall carriage rate of S.aureus,methicillin-resistant S.aureus (MRSA) and multi-drug-resistant S.aureus (MDRSA) among 1 012 primary school students were 40.1％,1.2％ and 4.0％,respectively.Most S.aureus isolates were resistant to penicillin,erythromycin and clindamycin.The dominant multidrug resistance patterns of MDRSA isolates were resistant to erythromycin-clindamycin-tetracycline and erythromycin-clindamycin-cefoxitin.Multifactor dimensionality reduction analysis showed that the rate of resistance to cefoxitin,tetracycline and chloromycetin among MDRSA was 104.39 times as much as that of nonMDRSA.Conclusions The carriage rate of S.aureus in healthy primary school students from Guangzhou was high and these isolates showed multidrug resistance.These data provide basis for guiding the rational use of antibiotics.

In the present study,we examined the codon usage bias between pseudorabies virus (PRV) US1 gene and the US1-like genes of 20 reference alphaherpesviruses.Comparative analysis showed noticeable disparities of the synonymous codon usage bias in the 21 alphaherpesviruses,indicated by codon adaptation index,effective number of codons (ENc) and GC3s value.The codon usage pattern of PRV US1 gene was phylogenetically conserved and similar to that of the US1-like genes of the genus Varicellovirus of alphaherpesvirus,with a strong bias towards the codons with C and G at the third codon position.Cluster analysis of codon usage pattern of PRV US1 gene with its reference alphaherpesviruses demonstrated that the codon usage bias of US1-like genes of 21 alphaherpesviruses had a very close relation with their gene functions.ENc-plot revealed that the genetic heterogeneity in PRV US1 gene and the 20 reference alphaherpesviruses was constrained by G+C content,as well as the gene length.In addition,comparison of codon preferences in the US1 gene of PRV with those of E.coli,yeast and human revealed that there were 50 codons showing distinct usage differences between PRV and yeast,49 between PRV and human,but 48 between PRV and E.coli.Although there were slightly fewer differences in codon usages between E.coli and PRV,the difference is unlikely to be statistically significant,and experimental studies are necessary to establish the most suitable expression system for PRV US1.In conclusion,these results may improve our understanding of the evolution,pathogenesis and functional studies of PRV,as well as contributing to the area of herpesvirus research or even studies with other viruses.

OBJECTIVE:To optimize the extraction technology of Zishen yangyin granules.METHODS:The yield of the extractum and the content of the extraction using 80% alcohol as solvent were used as indexes to optimize the extraction technology of Zishen yangyin granules by orthogonal experiment with the times of extraction,the extraction time and the amount of water added as factors.RESULTS:The optimal extraction conditions were as follows:the extraction was carried out for 3 times by adding water at an amount of 9 times,6 times and 6 times,respectively that of solid substance with decoction time of 3 h,2 h and 1 h,respectively.CONCLUSION:The optimal extraction process was proved to be time-saving and energy-saving and suitable for large production.

Objective:To study the morphologic standard values of craniofacial hard-tissue of the youths in Xi'an.Methods:CBCT scanned cephalometric data of 100 selected volunteers (50 males and 50 females)with individual normal occlusion were collected.31 landmarks and 31 measurements were compared between sexes and between 3D and 2D data with software InvivoDental 5.2,WinCeph 8.0.and SPSS 19.0.Results:1.In the 3D measurements,vertical growth of mandible in the females was more than that in the males. The values of torque of lower incisor,basis length,height of rumi mandibulae and length of corpora mandibulae in the males were bigger than those in the females.2.Compared with 2D measurements,there existed statistically significant differences in most parameters except U1-NA(mm).Conclusion:3D analysis with CBCT may provide more accurite morphologic data for craniofacial hard tissues.

Pyruvate phosphate dikinase (PPDK; EC 2.7.9.1) is found in certain microorganisms and plants, and catalyzes the conversion of AMP, PPi and phosphoenolpyruvate (PEP) to ATP, Pi and pyruvate. Using the genomic DNA of Microbispora rosea subsp. aerata as the template, a DNA fragment encoding the gene PPDK was amplified by PCR and inserted into the expression vector pET28a(+), yielding pET28a (+)-PPDK. The E. coli BL21 (DE3) was transformed with the pET28a (+)-PPDK. After inducing with IPTG, the E. coli BL21 (DE3) [pET28a (+)-PPDK] expressed recombinant PPDK fused to an N-terminal sequence of 6-His Tag. The molecular weight of PPDK was estimated to be 101 kD by SDS-PAGE. The PPDK was purified by His * Bind Resin affinity chromatography and ultrafiltration using 10 kD cut-off membrane. The successful application of PPDK in pyrosequencing was also demonstrated.
Actinomyces ; enzymology ; Escherichia coli ; genetics ; metabolism ; Pyruvate, Orthophosphate Dikinase ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Sequence Analysis