镇江地区慢性牙周炎与牙龈卟啉单胞菌寄生相关性研究

目的:建立基于Real-timePCR技术定量检测牙周组织中牙龈卟啉单胞菌的方法,并研究该细菌寄生与牙周炎严重程度的关系。方法:选择50例慢性牙周炎患者,包括重度牙周炎患者30例和轻度牙周炎患者20例;另选取牙周健康者20例。采集每位受试者龈沟液并提取细菌基因组DNA,采用Real-timePCR法检测样本中牙龈卟啉单胞菌数量。结果:重度牙周炎患者探诊深度≥7mm,轻度牙周炎患者探诊深度介于3mm至7mm之间,健康者探诊深度3mm。龈沟液中牙龈卟啉单胞菌数量随着牙周炎加重而显著增加(P0.05)。结论:牙龈卟啉单胞菌寄生数量与慢性牙周炎严重程度密切相关,可以反映牙周炎的严重程度。Real-time PCR法简单方便,可以快速检查牙龈卟啉单胞菌感染情况。     

表达Panton-Valentine杀白细胞素(PVL)的金黄色葡萄球菌诱导人THP-1单核细胞自噬和凋亡

目的 研究表达Panton-Valentine杀白细胞素(PVL)的金黄色葡萄球菌诱导人THP-1细胞自噬和凋亡机制.方法 在大肠杆菌中表达重组杀白细胞素(rPVL),并免疫家兔制备多克隆抗血清.采用反转录PCR和Western blot法鉴定表达PVL的耐甲氧西林金黄色葡萄球菌(MRSA).分别使用表达PVL的MRS...     

某院ICU医院获得性肺炎患者痰分离MRSA耐药基因和pvl基因携带情况

目的了解医院获得性肺炎患者痰中耐甲氧西林金黄色葡萄球菌(MRSA)耐药基因和毒力因子pvl基因携带情况。方法对来源于某院重症监护病房(ICU)医院获得性肺炎患者痰中的46株MRSA,采用聚合酶链反应(PCR)检测细菌耐药基因(mecA、aacA-D、tetK、tet M、msrA、msrB、ermA、ermC、vatA、vatB、vatC、femB和linA)和毒力因子pvl基因,并以PCR法分析MRSA菌株SCCmec型别。结果 46株MRSA中,耐药基因mecA、aacA-D、tetK、msrA、ermA、ermC、femB和linA的检出率分别为100%、54.35%、36.96%、13.04%、36.96%、52.17%、71.74%和10.87%,所有菌株均未检出tet M、msrB、vatA、vatB和vatC基因;毒力基因pvl携带率为65.22%。46株MRSA共检出4种SCCmec基因型,其中SCCmecⅡ型、Ⅲ型、IVc型、V型分别为26.09%、52.17%、2.17%和2.17%。结论医院获得性肺炎患者痰中MRSA携带多种耐药基因,且毒力基因pvl携带率较高,SCCmec基因型以Ⅲ型为主,临床医务人员对此情况应高度重视。     

肺炎患者痰液分离鲍曼不动杆菌耐药基因分析

目的:分析呼吸内科肺炎患者痰液分离鲍曼不动杆菌携带β-内酰胺酶基因及主动外排泵基因情况。方法:收集经生化鉴定为醋酸钙-鲍曼不动杆菌复合体的菌株160株,将鲍曼不动杆菌特异性片段(Ab-ITS)的二重PCR法确认为鲍曼不动杆菌者纳入研究。用PCR法检测鲍曼不动杆菌β-内酰胺酶基因和主动外排泵基因,包括A类β-内酰胺酶(blaTEM、blaPER和blaCARB)、B类β-内酰胺酶(blaIMP和blaVIM)、C类β-内酰胺酶(blaADC和blaDHA)、D类β-内酰胺酶(blaOXA-23、blaOXA-24、blaOXA-51和blaOXA-58)和主动外排泵相关基因(adeB、adeJ、macB、emrB、emrA、abeS、abeM和craA)。以微量肉汤稀释法检测加入外排泵抑制剂羰基氰化氯苯腙(CCCP)前后,携带adeB基因鲍曼不动杆菌对亚胺培南的最低抑菌浓度(MIC)的变化。结果:临床分离160株醋酸钙-鲍曼不动杆菌复合体中鲍曼不动杆菌143株(占90%),包括亚胺培南耐药株119株(占83.2%)和敏感株24株(占16.8%)。所有鲍曼不动杆菌均检出blaOXA-51基因,其中,亚胺培南耐药株中检出blaOXA-23(98.3%)、blaTEM(41.4%)、blaADC(98.3%)和blaDHA(0.7%),blaPER、blaCARB、blaIMP、blaVIM、blaOXA-24和blaOXA-58均未检出;敏感株中未检出blaOXA-23。adeJ、macB、abeM和craA基因在143株鲍曼不动杆菌中的携带率均为100%;emrB、emrA和abeS基因携带率均超过80%。adeB基因在亚胺培南耐药株中检出117株(98.3%)。在CCCP干预试验中,携带adeB基因的鲍曼不动杆菌MIC50降低1/2。结论:呼吸内科鲍曼不动杆菌主要携带blaOXA-23和adeB主动外排泵基因,可能与亚胺培南耐药密切相关。     

6对通用引物检测不同型别人乳头瘤病毒效果分析

目的: 比较6对通用引物检测9种不同型别人乳头瘤病毒（HPV）的效果,为临床提供一种快速简便的HPV检测方法。方法: 收集女性宫颈脱落细胞标本954例,使用HPV分型试剂盒进行HPV DNA检测并分型。分别使用6对通用引物（① GP5/GP6,② GP5+/GP6+,③ CP Ⅰ/CP ⅡS,④ CP I/CP ⅡG,⑤ 外：MY09/MY11,内：GP5+/GP6+,⑥ SPF1/GP6++）对HPV-DNA阳性标本进行PCR检测,比较6对通用引物检测9种不同类型HPV DNA效果。结果： 采用HPV分型试剂盒从954例标本中共检出263例HPV-DNA阳性标本,阳性率为27.57%。在阳性标本中,采用通用引物SPF1/GP6++检出的阳性例数为223例,检出率为84.79%;采用巢氏PCR引物（外：MY09/MY11,内：GP5+/GP6+）检出的阳性例数为207例,检出率为78.71%。上述两对通用引物均可以检出9种型别HPV DNA。结论： 通用引物SPF1/GP6++检测效果最佳,但引物含次黄嘌呤,成本较高;巢氏PCR引物（外：MY09/MY11,内：GP5+/GP6+）检测效果次之,且合成成本较低。在HPV研究中可以根据实际情况选用。     

我院鲍曼不动杆菌携带blaOXA-23基因及其耐药机制研究

目的研究我院分离鲍曼不动杆菌携带bla_OXA-23基因的携带情况及其诱导细菌耐药机制。方法收集江阴市青阳医院2019年1月至2019年12月间临床分离鲍曼不动杆菌,经VITEK-2全自动微生物鉴定及药敏分析系统鉴定耐亚胺培南菌株100株,敏感菌株100株。通过多重PCR法检测上述菌株中bla_OXA-23、bla_OXA-24、blaOXA-51和bla_OXA-58基因携带情况。通过PCR法扩增亚胺培南耐药的鲍曼不动杆菌bla_OXA-23的ORF序列,构建过表达载体bla_OXA-23/pYMAb3并电转化鲍曼不动杆菌(ATCC 17978),以硫酸卡那霉素和美罗培南共同筛选过表达bla_OXA-23基因的菌株。采用稀释法检测转化bla_OXA-23/pYMAb3载体和pYMAb3空载体的菌株对亚胺培南的MIC。结果我院耐亚胺培南的鲍曼不动杆菌均携带bla_OXA-23,敏感株未检出携带该基因。RT-PCR法可检出转化bla_OXA-23/pYMAb3载体菌株表达bla_OXA-23基因,而转化pYMAb3空载体菌株未检出bla_OXA-23基因表达。转化bla_OXA-23/pYMAb3载体菌株对亚胺培南MIC为32μg/mL,转化空载体菌株MIC仅为0.5μg/mL。结论我院耐亚胺培南的鲍曼不动杆菌均携带bla_OXA-23基因,可以作为耐药菌株筛选的分子标记。由质粒携带的bla_OXA-23基因是诱发我院鲍曼不动杆菌耐药的重要原因,在院感工作中需高度重视。     

鲍曼不动杆菌荚膜通过抑制PI3K/Akt/mTOR信号通路促进鼠巨噬细胞自噬和凋亡

[Abstract\]Objective: To study the capability of Acinetobacter baumannii with different thickness capsules induce Raw 264.7 cell autophagy and apoptosis, and its potential mechanisms. Methods: A baumannii was isolated from the sputum of patients with pneumonia. The bacterial capsule was stained by Congo red staining, and the bacterial strains with significantly different capsule thickness were selected for subsequent study. Raw 264.7 cells were infected with thick capsular strain (Ab H strain) or thin capsular strain (Ab B strain), and the cells were harvested at 1 h, 3 h and 6 h after infection. The expression of autophagy related proteins LC 3 and Beclin 1, as well as the phosphorylation levels of PI3K, Akt and mTOR were determined by Western blotting. Flow cytometry was used to determine the reactive oxygen species(ROS) production and apoptosis rate. Results: After A. baumannii was stained with Congo red, the cells were shown purple black, the background was red, and the capsule was not stained. The bacterial capsule thickness was clearly judged. Two strains with significant differences in capsular thickness were grouped, namely thick capsular strain (Ab H strain) and thin capsular strain (Ab B strain). The expression of autophagy related protein Beclin 1 of Raw 264.7 cells induced by Ab H strain were significantly higher than that of Ab B strain (P<0.05), and the apoptosis rate of Raw 264.7 cells induced by Ab H strain was also significantly higher than that of Ab B strain (P<0.05). Two strains of A. baumannii could induce a significant increase in the production of ROS in Raw 264.7 cells (P<0.05), and the ability of Ab H strain to induce ROS production was significantly higher than that of Ab B strains (P<0.05). After Raw 264.7 cells were infected with two strains respectively, the phosphorylation levels of PI3K, Akt and mTOR were significantly inhibited, and the inhibition increased when the time prolonged (P<0.05). Conclusion: A. baumannii could induce autophagy and apoptosis of Raw 264.7 cells, and the induction ability is positively correlated with bacterial capsule thickness. A. baumannii with thick capsule has a stronger ability to induce ROS production in Raw 264.7 cells and inhibit the activation of PI3K/Akt/mTOR signaling pathway, which ultimately enhances autophagy and apoptosis of Raw 264.7 cells.     

肺炎克雷伯菌所致肝脓肿患者的临床特征及毒力基因检测

目的 分析肺炎克雷伯菌肝脓肿(KPLA)患者的临床特征,了解肺炎克雷伯菌毒力基因携带情况,为临床早期诊断和合理治疗提供参考.方法 回顾性分析江苏大学附属医院2017年7月—2019年8月收治的34例脓液培养阳性的细菌性肝脓肿患者的临床资料,分为KPLA组和非肺炎克雷伯菌肝脓肿(NKPLA)组.采用VITEK 2 Com...     

3种主要短链脂肪酸减轻5-氟尿嘧啶诱发的THP-1细胞炎症反应

目的:研究3种主要短链脂肪酸(short-chain fatty acids,SCFAs)即乙酸钠、丙酸钠和丁酸钠,对5-氟尿嘧啶(5-Fluorouracil,5-FU)诱发人单核巨噬THP-1细胞炎症反应的影响及其可能机制.方法:将THP-1细胞分为5组:对照组,细胞未经任何处理;5-FU组,2.5 mmol/L ...     

Ferroportin在前列腺癌骨转移中的表达及意义

目的分析Ferroportin在前列腺癌骨转移中的表达,探讨Ferroportin在评估前列腺癌发生和进展中的意义。方法取行前列腺穿刺活检的90例患者,根据组织病理和全身骨扫描结果,分为前列腺癌骨转移组(PM组)30例,前列腺癌无骨转移组(NPM组)30例和前列腺增生组(BPH组)30例。检测血常规、血生化,并对前列腺穿刺活检组织采用Real-time PCR和免疫组化技术检测Ferroportin表达,分析各组间表达差异。结果前列腺癌骨转移组Ferroportin基因表达量较其他组明显下降(P0.05);前列腺癌组织中Ferroportin表达量较前列腺组织明显减少,差异明显;Ferroportin的表达和前列腺癌的恶性程度呈负相关。结论 Ferroportin可作为预测前列腺癌发生和进展指标,并可能是前列腺癌靶向治疗的新靶点。