本文選題：高毒力肺炎克雷伯菌 + 分子特征��； 參考：《南昌大學(xué)》2017年碩士論文

【摘要】：目的:1.研究高毒力肺炎克雷伯菌血流感染的流行特征及危險(xiǎn)因素分析,為臨床合理治療和防治高毒力肺炎克雷伯菌血流感染提供科學(xué)依據(jù)。2.探究血流感染中高毒力肺炎克雷伯菌毒力、分子流行病學(xué)特征及其對(duì)中性粒細(xì)胞功能的影響。3.初步探究高毒力肺炎克雷伯菌延遲中性粒細(xì)胞凋亡的調(diào)控機(jī)制。方法:1.收集南昌大學(xué)第一附屬醫(yī)院2014年1月-2016年12月住院患者血液標(biāo)本分離出的肺炎克雷伯菌178株。采用腸桿菌科基因間重復(fù)一致序列聚合酶鏈反應(yīng)(ERIC-PCR)與多位點(diǎn)序列分析法(MLST)確定主要基因分型及其病區(qū)分布。2.聚合酶鏈?zhǔn)椒磻?yīng)(PCR)及測(cè)序技術(shù)確定其血清莢膜分型及毒力基因攜帶,本研究定義凡是符合以下任何一項(xiàng):攜帶K1、K2、rmpA/rmpA2基因或者高黏液表型陽(yáng)性的菌株為高毒力菌株,否則為非高毒力菌株;查詢(xún)統(tǒng)計(jì)分析這178例患者的臨床資料,實(shí)驗(yàn)分為高毒力菌株組與非高毒力菌株組,運(yùn)用單因素分析和多因素logistic回歸分析高毒力肺炎克雷伯菌血流感染的危險(xiǎn)因素。3.收集南昌大學(xué)第一附屬醫(yī)院2016年6月-2016年12月住院患者血標(biāo)本分離出的肺炎克雷伯菌55株,PCR擴(kuò)增及測(cè)序確定其wzi分型,PCR擴(kuò)增確定其ompK36等位基因分群(A、B、C、D),用血清抗性、抗中性粒細(xì)胞吞噬試驗(yàn)來(lái)研究肺炎克雷伯菌的體外毒力性;采用流式細(xì)胞儀檢測(cè)患者外周血中性粒細(xì)胞表面分子表達(dá)的變化,采用ELISA檢測(cè)患者體內(nèi)炎癥因子的釋放量。4.建立體外中性粒細(xì)胞感染模型和體內(nèi)腹腔感染膿毒癥動(dòng)物模型,采用Western-Blot檢測(cè)中性粒細(xì)胞凋亡調(diào)控蛋白的表達(dá)情況。結(jié)果1.178株血流感染肺炎克雷伯菌,主要基因分型是ST11型,其中HvKP菌株以ST23型為主。主要見(jiàn)于消化科、燒傷科、神經(jīng)外科、重癥醫(yī)學(xué)科,且呈現(xiàn)克隆性傳播。2.伴有糖尿病基礎(chǔ)疾病、引流、血型(A型)、使用喹諾酮類(lèi)及碳青霉烯類(lèi)抗菌藥物是HvKP血流感染的獨(dú)立危險(xiǎn)因素。3.在55株血流感染KP中,有20株HvKP菌株,35株non-HvKP菌株。HvKP菌株的MLST分型以ST23和ST65為主,而non-HvKP菌株則以ST11型為主;HvKP菌株的ompK36等位基因以C群為主,而non-HvKP菌株則以A群和D群為主;HvKP菌株的莢膜血清分型以wzi2-K2和wzi1-K1為主,而non-HvKP菌株則以wzi64-K64分型為主。4.在20株血流感染HvKP中,有16株表現(xiàn)出高水平的血清抗性能力,而在35株non-HvKP中,僅有4株表現(xiàn)出高水平的血清抗性能力;HvKP菌株攜帶毒力相關(guān)基因數(shù)多于non-HvKP菌株。5.在抗中性粒細(xì)胞吞噬方面,大部分non-HvKP菌株的中性粒細(xì)吞噬率均在55-80%左右,而大部分HvKP菌株的中性粒細(xì)胞吞噬率在30%左右。6.相比較non-HvKP組,HvKP組患者體內(nèi)PD-1、CD86表達(dá)明顯下調(diào),而PD-L1、CD80明顯變化;HvKP組患者體內(nèi)IL-2、IL-8、IL-1β釋放量明顯增多,而TNF-α釋放量明顯降低。7.在HvKP感染的早期,HvKP感染組NF-κB P65蛋白含量顯著高于non-HvKP感染組;HvKP感染組Bax/Bcl-2的比值顯著低于non-HvKP感染組;HvKP感染組Caspase-3,8,9蛋白含量顯著低于non-HvKP感染組。結(jié)論:1.在本院血流感染KP菌株的MLST分型以ST11為主,而血流感染HvKP菌株以ST23、ST65型為主。2.通過(guò)本文對(duì)HvKP血流感染的危險(xiǎn)因素分析,提示我們要特別重視合并有慢性?xún)?nèi)科基礎(chǔ)疾病,特別是伴有糖尿病的血流感染KP患者;其次要注意引流操作的無(wú)菌意識(shí)及引流時(shí)間和碳青霉烯類(lèi)、喹諾酮類(lèi)抗菌藥物的合理使用。3.HvKP菌株的ompK36等位基因以C群為主,莢膜血清分型以wzi2-K2和wzi1-K1為主。4.相比較non-HvKP菌株,HvKP菌株攜帶毒力相關(guān)基因數(shù)較多,血清抗性能力和對(duì)中性粒細(xì)胞吞噬能力均較高。5.HvKP血流感染患者體內(nèi)炎癥因子的釋放量改變以及PMN表面分子的表達(dá)紊亂,提示HvKP感染可能使PMN的功能發(fā)生了變化。6.我們推測(cè)在HvKP感染的早期,HvKP誘導(dǎo)PMN凋亡延遲的作用可能是通過(guò)調(diào)節(jié)Bax/Bcl-2比值使Caspase-3,8,9的表達(dá)下調(diào),進(jìn)而激活NF-κB,從而延遲中性粒細(xì)胞的凋亡。
[Abstract]:Objective: 1. to study the epidemiological characteristics and risk factors of high virulence Klebsiella pneumoniae blood flow infection and to provide scientific basis for the rational treatment and prevention of Klebsiella pneumoniae blood flow infection by.2. to explore the virulence of Klebsiella pneumoniae, the characteristics of molecular epidemic and the effect on neutrophils function in the blood flow infection. .3. preliminarily explored the regulatory mechanism of delayed neutrophils apoptosis of Klebsiella pneumoniae. Methods: 1. collect 178 Klebsiella pneumoniae isolated from the blood specimens of hospitalized patients in the First Affiliated Hospital of Nanchang University in December January 2014. Using INTERGENE repeat sequence polymerase chain reaction (ERIC-PCR) and multiple sites in the Enterobacteriaceae. The point sequence analysis (MLST) determines the main genotyping and its endemic area distribution.2. polymerase chain reaction (PCR) and sequencing technology to determine its sera pods and virulence genes. This study defines any of the following: strains with K1, K2, rmpA/rmpA2, or high mucous phenotypes are highly virulent, otherwise Non high virulence strains; the clinical data of these 178 patients were analyzed and analyzed. The experiment was divided into high virulent and non virulence strains. Single factor analysis and multiple factor Logistic regression analysis were used to analyze the risk factor of Klebsiella pneumoniae blood flow infection.3.. The First Affiliated Hospital of Nanchang University was hospitalized in December -2016 June 2016. 55 strains of Klebsiella pneumoniae isolated from blood samples, PCR amplification and sequencing to determine their WZI typing, and PCR amplification to determine their ompK36 allele groups (A, B, C, D). Serum resistance and anti neutrophil phagocytosis test were used to study the virulence of Klebsiella pneumoniae in vitro. Flow cytometry was used to detect the surface of neutrophils in peripheral blood of patients. The changes in the expression of the sub expression, using ELISA to detect the release of inflammatory factors in the patient's body, to establish an in vitro neutrophils infection model and an animal model of sepsis in the body, and to detect the expression of the regulation of the apoptosis of neutrophils by Western-Blot. Results 1.178 strains of Klebsiella pneumoniae were infected with the main genotyping of ST1. Type 1, of which HvKP strain is mainly ST23, mainly in the digestive, burn, Department of Neurosurgery, and severe medical sciences, and presents clonically transmitted.2. with diabetes based diseases, drainage, blood group (A), the use of quinolones and carbapenems as an independent risk factor for HvKP blood flow infection,.3. in 20 blood stream infection KP, there are 20 strains of blood flow infection. The MLST genotyping of 35 strains of non-HvKP strain.HvKP was dominated by ST23 and ST65, while the non-HvKP strain was dominated by ST11 type, and the ompK36 allele of the HvKP strain was dominated by C group, while the non-HvKP strain was the dominant group. Of the main.4. in 20 blood flow infection HvKP, 16 showed high level of serum resistance, but only 4 of 35 strains of non-HvKP showed high level of serum resistance. The number of Virulence Related Genes of strain HvKP was more than that of non-HvKP strain.5. in the anti neutrophil phagocytosis, and the neutrophil phagocytosis rate of most of the strains of non-HvKP strains were all At about 55-80%, the neutrophil phagocytosis rate of most HvKP strains was compared to that of non-HvKP group at about 30%.6., and PD-1, CD86 expression in HvKP group was obviously down, while PD-L1 and CD80 were obviously changed, IL-2, IL-8, and beta release in the HvKP group were significantly increased. The content of NF- kappa B P65 protein was significantly higher than that in non-HvKP infection group, and the ratio of Bax/Bcl-2 in HvKP infection group was significantly lower than that in non-HvKP infection group, and the Caspase-3,8,9 protein content in HvKP infection group was significantly lower than that in non-HvKP infection group. Conclusion: 1. Through the analysis of the risk factors of HvKP blood flow infection, we suggest that we should pay special attention to the patients with chronic basic diseases, especially the KP patients with blood flow infection with diabetes; secondly, we should pay attention to the aseptic consciousness of the drainage operation, the time of drainage and the carbapenems, the rational use of the ompK36 of the.3.HvKP strain of the quinolone antibiotics. The allele was mainly C group, and the capsule serotyping was compared with the non-HvKP strain of wzi2-K2 and wzi1-K1 as the main.4. phase. The HvKP strains carried more virulence related genes. The serum resistance ability and the neutrophil phagocytosis were all higher in the patients with.5.HvKP blood flow infection, and the expression of PMN surface molecules was disordered. It is suggested that HvKP infection may change the function of PMN.6.. We speculate that in the early stage of HvKP infection, HvKP induced PMN apoptosis delayed by regulating the Bax/Bcl-2 ratio to decrease the expression of Caspase-3,8,9, and then activate NF- kappa B to delay the apoptosis of neutrophils.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R446.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 楊志剛;馬希濤;雷小莉;;醫(yī)院感染多藥耐藥菌的危險(xiǎn)因素分析[J];中華醫(yī)院感染學(xué)雜志;2014年04期

2 吳國(guó)榮;陳國(guó)千;王春新;耿先龍;李一龍;楊小娟;;重癥監(jiān)護(hù)病房患者中性粒細(xì)胞VCS參數(shù)檢測(cè)的臨床意義[J];中華醫(yī)院感染學(xué)雜志;2011年19期

3 梁俊容;景懷琦;;耶爾森菌抵抗宿主免疫并誘導(dǎo)吞噬細(xì)胞凋亡機(jī)制研究[J];中國(guó)人獸共患病學(xué)報(bào);2011年08期

4 姜麗娜;李柏青;姚春艷;金齊力;;Th1型和Th2型細(xì)胞因子對(duì)人中性粒細(xì)胞吞噬結(jié)核桿菌活性的影響[J];中華微生物學(xué)和免疫學(xué)雜志;2010年06期

5 王曉龍;周向東;;核轉(zhuǎn)錄因子-κB抑制因子對(duì)中性粒細(xì)胞凋亡的影響[J];中國(guó)老年學(xué)雜志;2010年02期

6 張國(guó)明;王禹;李天德;張大為;劉秀華;楊菲菲;;應(yīng)激活化蛋白激酶在大鼠缺血后適應(yīng)中的變化及其對(duì)細(xì)胞凋亡影響的研究[J];浙江大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2009年06期

7 王玲;劉麗宏;單保恩;張超;桑梅香;李嘉;;Celecoxib下調(diào)NF-кB信號(hào)通路促進(jìn)人乳腺癌MDA-MB-231細(xì)胞凋亡的體外研究[J];癌癥;2009年06期

8 李娟;王曉輝;呂曉菊;;成人肺炎克雷伯菌敗血癥53例臨床分析[J];中國(guó)抗生素雜志;2008年08期

9 鄧雪松;倪勇;高海斌;王成友;;膽道梗阻大鼠中性粒細(xì)胞凋亡與IL-8水平的關(guān)系[J];嶺南現(xiàn)代臨床外科;2008年02期

10 甄作均;陳煥偉;崔偉珍;張海雄;楊少儀;;細(xì)菌性肝膿腫62例診治經(jīng)驗(yàn)分析[J];中國(guó)實(shí)用外科雜志;2007年12期

，

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