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The Study on Sanhuang Capsule to Eliminate R Plasmid of Neisseria Gonorrhoeae

Author: YeYing
Tutor: LinZhaoChun；YongGang
School: Luzhou Medical College
Course: Dermatology and Venereology
Keywords: Neisseria gonorrhoeae Plasmid Three yellow capsules
CLC: R285.5
Type: Master's thesis
Year: 2009
Downloads: 52
Quote: 0
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Abstract


The purpose of the bacterial plasmid including the F factor (fertility factor), R factor (re--sistance factor) the Col factor (coliconogenic factor), Ti factors (tumor inducing plasmid), huge the plasmid (mega plasmid) and degradation plasmid. R factor, also known as R plasmid R plasmid carrying the resistance gene, given the resistance of bacteria to certain antibiotics. In the past 30 years, some domestic and foreign researchers actively looking for an effective way to eliminate the resistance plasmid reverse bacterial resistance. Gonorrhea is one of the most common sexually transmitted disease, gonorrhea the sergenti resistance bacteria resistant With the widespread use of antibiotics, more and more serious. Numerous studies found resistant Neisseria gonorrhoeae is mainly mediated by the plasmid. The useful domestic traditional Chinese medicinal plasmid elimination of the G-bacteria, the kind of traditional Chinese medicine is just the prescription of Chinese medicine to treat gonorrhea, we can speculate that the basis of traditional Chinese medicine treatment of gonorrhea and plasmid eliminate contact. This study was to study three yellow capsule is to eliminate the role of R plasmids in Neisseria gonorrhoeae, as well as the duration of action of the three yellow capsules extend the ability to enhance the consumer quality effects. Method 1. Neisseria gonorrhoeae plasmid extraction: Neisseria gonorrhoeae selection of 10 by the oxidase test and sugar fermentation tests identified in 2008 at the Sichuan Institute of Dermatology and Venereology, outpatient patient separation, first select 10 TM medium recovery 24 hours after the transfer species in 10 TM medium, wait 24 hours and then ring vaccination were scraped Neisseria gonorrhoeae 10 medium 10 Sheng 1ml saline Ep tube and marked with a number, it gently pipetting broth mix with the pipette. Ep tube broth extracted in accordance with the AxyPrep plasmid box manual steps to extract plasmid extraction, after conclusion of the 10 Neisseria gonorrhoeae plasmid extract were added to the bromophenol blue dye stained pipette plasmid extraction The liquid and DNAmarker (300bp-10000bp) Add to the prepared 100v electrophoresis in 2% agarose gel-like hole 40min the plasmid strip and make a record and then observed under a light in the ultraviolet perspective. 2 yellow capsule sub-inhibitory concentration determination: the turn of the 10 Neisseria gonorrhoeae strains recovery species with ring vaccination were scraped a little strain in 10 1ml saline containing glass tubes will its mix and placed in a densitometer and adjust the amount of bacteria and saline amount of its optical density of 0.3, thereby preparing a bacterial concentration of 105/ml bacterium Neisseria gonorrhoeae. The broth inoculated with inoculator containing different amounts (6.25ul 12.5ul, 25ul, 50ul, 100ul, 200ul) the three yellow capsule suspension TM medium (medium amount of each dish 15ml) its growth was observed after 24 h, in order to determine its sub-inhibitory concentrations, thereby the preparation containing sub-inhibitory concentrations of three yellow capsule suspension TM medium. Neisseria gonorrhoeae plasmid eliminate experiment: recovery above 10 Neisseria gonorrhoeae same way again, transferred species turn again after 24h species in three yellow capsule containing sub-inhibitory concentrations suspension of TM training base, respectively. role 24h, 48h, 72h after extracting plasmid electrophoresis observation plasmid with changes every 24h turn all kinds of time, the specific operation is as follows: after 24h of Neisseria gonorrhoeae will turn species inoculated in containing sub-inhibitory concentrations three yellow capsule suspension TM medium cultured for 24 hours in accordance with the first method of extracting plasmid extracted plasmid and electrophoresis consumer matter before electrophoresis results for comparison, Neisseria gonorrhoeae turn again containing three species in the yellow the capsules suspension of TM medium, 24 hours later, the same method to extract plasmid electrophoresis consumer quality before electrophoresis results for comparison, The Neisseria gonorrhoeae turn again to species containing three yellow capsule suspension TM medium, after 24 hours extraction plasmid electrophoresis and elimination qualitative results contrast. Compared to the last three by three yellow capsule role 24h, 48h, 72h electrophoresis results. Results all enrolled Neisseria gonorrhoeae are growing well, and is not to be contaminated. 1 extract plasmid overview: 10 gonorrhea resistant sergenti bacteria in nine visible clearer plasmid band, and its molecular weight both 7.4Kb Asian type PPNG strains. 2. Of gonorrhea resistant Joseph's bacteria containing different amounts (6.25ul, 12.5ul, 25ul, 50ul, 100ul, 200ul) the three yellow capsule suspension TM medium (medium amount of each dish 15ml) Neisseria gonorrhoeae can be observed after 24h culture growth of the three yellow capsules subinhibitory concentrations (i.e. make the maximum drug concentration in the growth of Neisseria gonorrhoeae), identified as 200ul / dish, this concentration eliminate interstitial fluid is the experiment with consumer quality agent. Observed after plasmid band situation (with 3.10 Neisseria gonorrhoeae were grown in medium containing three yellow capsule ingredients TM 24h, 48h, 72h, were extracted three time points Neisseria gonorrhoeae plasmid electrophoresis eliminate quality compared to before): after 24h plasmid bands decreased by 2; 48h after a decrease of 3; 72h after a decrease of 8. Chi-square test consumer quality agent role 24h, 48h Neisseria gonorrhoeae plasmid negative rate and eliminate quality Neisseria gonorrhoeae plasmid negative rate no significant differences in sex (P gt; 0.05), eliminate quality agents for 72h Neisseria gonorrhoeae bacteria plasmid negative rate and eliminate quality Neisseria gonorrhoeae plasmid negative rate significant differences (P lt; 0.05). The chi-square test 24h, 48h, 72h three the point elimination qualitative rate significant differences (P lt; 0.05), its cancellation qualitative effect extended time with the role of the consumer quality and more obvious. The conclusion three yellow capsules eliminate quality, of its elimination qualitative effect with time and more obvious that the highest rate of 72-hour cancellation quality.

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