Experimental Study On The Reversal Effect Of Drug - Proliferation Of ESBLs - Producing Escherichia Coli By Purple Evil Theory

Background With the sensitivity of Escherichia coli (E.coli) to ultra broad spectrum antibiotics is declining, the spread of bacterial drug resistance of E. coli calls for a greater attention. Antibiotic resistant strains of Escherichia coli often carry multiple drug-resistant genes such as aminoglycoside resistant genes, quinolone resistance genes, chloramphenicol resistance genes and sulfadiazine resistance genes. It is often associated with a variety of mechanisms of drug resistance, and possess the features of multidrug resistance. It is urgent to find an effective therapy strategy to treat the infection, reduce the selective pressure of antibiotics, inhibit and reverse the drug resistance of Escherichia coli. Traditional Chinese medicine treatment has a definite effect on infection, and it is less susceptible to drug resistance due to its benign regulative effect. Traditional Chinese medicine has made considerable achievements in inhibiting bacterial resistance and restoring the sensitivity of resistant bacteria. My mentor advanced a theory that there is corresponding nature between drug-resistant bacteria infection and traditional Chinese medicine "Fuxie" theory, according to his view of drug-resistant bacteria infection and years of clinical observation. He proposed refreshing method in the treatment of drug-resistant bacteria infection, which avert the side effects due to abuses of drugs of cold nature. My mentor integrated the therapeutic advantage of TCM in exogenous febrile disease and epidemic febrile disease. He composed an improved decoction "Xin Jia Da Yuan San", which originated from the representative decoction in refreshing method-"Da Yuan Yin", and obtained good clinical effect. Therefore, we put forward the hypothesis that the refreshing method based on the theory of "Fuxie" can reverse bacterial resistance.Objective To study the effect of "Xin Jia Da Yuan San" Decoction Medicated serum on the antibiotic resistance of E. coli which generates extended-spectrum-lactamases and the influence of different substrates on the reversal effect in vitro intervention experiment.Method Serum pharmacology method is applied to prepare "Xin Jia Da Yuan San" Decoction medicated serum to simulate the actual compounds and interacting environment. Qualitatively analysis is used to detect the predicted active ingredients baicalin and paeoniflorin in meditated serum. Suspicious multi-drug-resistant Escherichia coli strains are isolated from clinical sputum samples. Automated microbial identification and drug sensitivity analysis system are used to determine bacteria identification and analyze drug susceptibility. Drug-resistant E.coli which generates extended-spectrum-lactamases is screened and confirmed by Kirby-Bauer test recommended by CLSI to be chosen as the experimental bacteria. In order to observe the bacteriostatic effect of serum itself, the experimental strains are cultured with linear incremental additive amount of serum mixed with broth. The experimental strains should be cultured with the additive amount of serum that under the level of displaying bacteriostatic in the intervention experiment. K-B disk diffusion method are used to measure the diameter of inhibition zone in different cultivate time with substrates ceftazidime and cefotaxime. Normal control group is blank serum group. The differences of bacterial drug resistance are compared between the three groups, and the reverse rate of bacterial resistance is calculated.Results Drug resistance E.coli which generates extended-spectrum-lactamases was chosen as the experimental bacteria from Guanganmen Hospital of academy of Chinese traditional medical sciences. The bacteriostatic ability test showed that the maximum amount of medicated serum and blank serum is 80% and 90%. After the intervention experiment, the result showed that there were significant differences of the diameter of inhibition zone between medicated serum and blank serum(P<0.05), yet no significant differences between blank serum and blank broth(P>0.05). There were no significant differences of the diameter of inhibition zone in different cultivate time(P>0.05). The reverse rate of bacterial resistance tested by ceftazidime is 116%, higher than cefotaxime which is 100%.conclusion 1. the Drug-resistance of ESBLs-producing E.coli can be reversed to a certain extend by "Xin Jia Da Yuan San" Decoction medicated serum, and this reversal effect were stable in different cultivate time.2. Ceftazidime was the optimal substrate contrasting with ceftazidime for the experimental strain.