Study The Action Effect Of Holly Bark On Producing Extended Spectrum Beta-Lactamases (ESBLs) Escheriohia Coli And Pharmacokinetic Research

This research was aimed to investigate the bacteriostatic mechanism of the Holly Bark for producing extended spectrum beta-lactamases(ESBLs) Escheriohia coli and pharmacokinetic study in animal to provide reference for the research and development of Holly Bark. The main research as follows:1 Resistance gene types were determined by identification of bacteria isolated in clinics:ESBLs-producing Escheriohia coli was tested by double slips synergy, and its genotype was determined through PCR method. Result Escheriohia coli producing ESBLs and carrying multidrug fosfomycin resistance gene(fosA3) was isolated.2 Study on the effect of water extractions of Holly Bark on ESBLs-producing bacterial:Antibacterial activity and fungicidal activity of water extractions of Holly Bark were tested by oxford cup method and spread-plate method. The minimal inhibiton concertration(MIC) of water extractions of Holly Bark and antibacterial agents were tested through twice micro-dilution method. Fractional inhibitory concentration(FICI) indexes of water extractions of Holly Bark combined with antibacterial agents were determined by microdilution checkerboard techniques. Results the MIC of water extractions of Holly Bark was 0.5 g/mL. When application of water extractions of Holly Bark combined with Amoxicillin,Ciprofloxacin Norfloxacin, Mequindox,Clinafloxacin, Sulfamonomethoxine, Gatifloxacin, Ceftazidime the FICI were less than or equal to 0.5, showed an additive,with Levofloxacin, Amikacin the FICI were more than 0.5 and less than 1, showed an synergistic correlation. Research shows that water extractions of Holly Bark had some inhibitory activity and fungicidal effect on the ESBLs-producing Escheriohia coli. Antibacterial activities of antibacterial agents against the ESBLs-producing Escheriohia coli are enhanced significantly by water extractions of Holly Bark.3 Research on the effect of Holly Bark different extractions combined with antibacterial agents on ESBLs-producing bacterial:Four kinds of Holly bark extracts were prepared. Producing extended spectrum beta-lactamases(ESBLs) bacterial were induced and subcultured by the extracts of Holly Bark combined with antibacterial agents, through 96-well reaction plate twice micro-dilution method. Result four kinds of Holly bark extracts combined with antibacterial agents to induce and subculture bacteria passage for drug-resistant Escherichia coli had antibacterial activity a different extent, which extracts of butanol, ethanol combined with antibacterial agents were best effect. Research shows that Holly bark extracts of butanol, ethanol combined with antibiotics to induce bacterial passage had anti-drug-resistant effect in vitro.4 Study on the effect of the serum of Holly Bark Combined with antibacterial agents on ESBLs-producing bacterial:The serum of Holly Bark was prepared. The minimal inhibition concentration (MIC) of antimicrobials were tested by twice micro-dilution method.Extended spectrum ?-lactamases (ESBLs)-producing bacteria were induced and subcultured by the serum of Holly Bark combined with antibacterial agents. Result the serum of Holly Bark combined with antibacterial agents to induce and subculture bacteria passage for ESBLs-producing E.coli had different extent antibacterial activity. Research shows that the effects of (3-lactam antibiotics, quinolones, aminoglycosides and kaba oxygen against drug-resistant E.coli were obviously strengthened when combined with the serum of Holly Bark5 Research on the bacteriostatic mechanism of the water extracts of Holly Bark on ESBLs-producing bacterial:Study on the bacteriostatic mechanism of the water extracts of Holly Bark through the cell ultrastrueture was observed, the permeabilty of cell wall and membrane was changed, nucleic acid synthesis was inhibited, protein synthesiswas inhibited, SDS-polyacrylamide gel electrophoresis were researched in this study. Result shows compared with the control(no extracts),the water extracts of Holly Bark destroyed the cells seriously, permeability of cell membrane and wall were increased, cell components were released, bacterial liquid AKP and soluble protein content were increased, total protein content were decreased, bacterial liquid UV absorbing compoundswere decreased, DNA fluorescence intensity weakened obviously of E.coli. Research shows that the bacteriostatic mechanism of the water extracts of Holly Bark is based on its roles about change the cell wall and membrance permeability,inhibite nucleic and protein synthesis of bacterial.6 Pharmacokinetics of Holly bark investigated in animals:High performance liquid chromatography used in analysising syringin of chicken plasma were established, and pharmacokinetic were researched by intragastric admininistration. The results showed that the plasma concentration of syringin in chicken, the alibration curve was linear(R2=0.9997)with in the range of 0.50-30.00 ug/mL. The main pharmacokinetic parameters were as follow: AUC(572.775) h-ug/mL, t1/2?(16.75)h, Cmax(32.571) ug/mL. Study showed that the plasma concentration of syringin which is best fitted using non-compartment model and could be used for pharmacokinetic of syringin in chicken.