TLR4与慢性间歇性低氧诱导炎症反应机制的研究进展

慢性间歇性低氧(chronic intermittent hypoxia,CIH)是阻塞性睡眠呼吸暂停综合征(obstructive sleep apnea syndrome,OSAS)的典型生理病理特征之一,是导致OSAS多器官系统损伤的关键起始因素。而Toll样受体4(Toll-like receptor 4,TLR4)作为一种高度保守的免疫受体,参与多种炎症反应及炎症级联反应。近年来,TLR4信号通路在CIH诱导炎症反应中的作用已经引起了越来越多的关注和研究,为OSAS的诊断及治疗提供了新靶点。该文从CIH、TLR4、TLR4基因沉默实验及拮抗剂等方面的最新研究进展进行综述。     

A novel microbead-based microfluidic device for rapid bacterial identification and antibiotic susceptibility testing

Effective treatment of infectious diseases depends on the ability to rapidly identify the infecting bacteria and the use of sensitive antibiotics. The currently used identification assays usually take more than 72 h to perform and have a low sensitivity. Herein, we present a microbead-based microfluidic platform that is highly sensitive and rapid for bacterial detection and antibiotic sensitivity testing. The platform includes four units, one of which is used for bacterial identification and the other three are used for susceptibility testing. Our results showed that Escherichia coli O157 at a cell density range of 10¹–10⁵ CFU/μL could be detected within 30 min. Additionally, the effects of three antibiotics on E. coli O157 were evaluated within 4–8 h. Overall, this integrated microbead-based microdevice provides a sensitive, rapid, reliable, and highly effective platform for the identification of bacteria, as well as antibiotic sensitivity testing.     

NF-κB结合元件缺失对NOX1启动子转录活性的影响

目的:构建含NADPH氧化酶1(NOX1)近端启动子区的pGL3萤光素酶报告基因载体及缺失NF-κB结合元件的相应载体,分别测定其相应活性,探讨NF-κB结合元件缺失对NOX1启动子区转录活性的影响。方法:采用PCR法克隆NOX1启动子区序列(1 415 bp),将目的片段与pGL3萤光素酶载体分别双酶切、纯化后进行连接(pGL3-NOX1-1415),测序鉴定;应用Alibaba 2.1软件分析NOX1近端启动子区,获取NF-κB结合元件;重叠PCR法将含该元件的启动子区域(88 bp)缺失,并构建相应载体(pGL3-NOX1-1327)。将两载体分别与pRL-TK内参质粒瞬时转染进入A549细胞,采用TNF-α(10μg/L)刺激细胞24 h,萤光酶标仪检测A549细胞的萤光素酶活性。结果:测序鉴定结果提示pGL3-NOX1-1415及NF-κB结合元件缺失的pGL3-NOX1-1327载体构建成功;细胞实验显示,TNF-α刺激后,转染pGL3-NOX1-1415的A549细胞萤光素酶活性明显强于对照组(P0.05),而转染NF-κB结合元件缺失的pGL3-NOX1-1327的细胞萤光素酶活性明显低于转染pGL3-NOX1-1415组(P0.05)。结论:TNF-α诱导A549细胞NOX1基因活化与NF-κB密切相关,NF-κB参与了TNF-α诱导的NOX1基因启动子的转录调控。     

Preferential infiltration of interleukin‐4‐producing CXCR4+ T cells in the lesional muscle but not skin of patients with dermatomyositis

Dermatomyositis (DM) and polymyositis (PM) are collectively termed autoimmune myopathy. To investigate the difference between muscle‐ and skin‐infiltrating T cells and to address their role for myopathy, we characterized T cells that were directly expanded from the tissues. Enrolled into this study were 25 patients with DM and three patients with PM. Muscle and skin biopsied specimens were immersed in cRPMI medium supplemented with interleukin (IL)‐2 and anti‐CD3/CD28 antibody‐conjugated microbeads. The expanded cells were subjected to flow cytometry to examine their phenotypes. We analysed the cytokine concentration in the culture supernatants from the expanded T cells and the frequencies of cytokine‐bearing cells by intracellular staining. There was non‐biased in‐vitro expansion of tissue‐infiltrating CD4⁺ and CD8⁺ T cells from the muscle and skin specimens. The majority of expanded T cells were chemokine receptor (CCR) type 7^–CD45RO⁺ effecter memory cells with various T cell receptor (TCR) Vβs. The skin‐derived but not muscle‐derived T cells expressed cutaneous lymphocyte antigen (CLA) and CCR10 and secreted large amounts of IL‐17A, suggesting that T helper type 17 (Th17) cells may have a crucial role in the development of skin lesions. Notably, the frequency of IL‐4‐producing chemokine (C‐X‐C motif) receptor (CXCR)4⁺ Th2 cells was significantly higher in the muscle‐derived cells and correlated inversely with the serum creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) levels. stromal‐derived factor (SDF)‐1/CXCL12, a ligand for CXCR4, was expressed at a high level in the vascular endothelial cells between muscular fasciculi. Our study suggests that T cell populations in the muscle and skin are different, and the Th2 cell infiltrate in the muscle is associated with the low severity of myositis in DM.     

常见评分系统对社区获得性肺炎预后的效果评价

目的探讨CURB-65评分、A-DROP评分、q-SOFA评分和中性粒细胞/淋巴细胞比值(NLR)对社区获得性肺炎(CAP)死亡率的预测价值。方法回顾性分析2016年1月-2018年12月间安徽医科大学第二附属医院呼吸与危重医学科收治的196例CAP患者的临床资料,所有病例均符合纳入排除标准。入院后24 h内测量生命体征,并在抗生素治疗前收集患者静脉血,测定血清生化等指标,通过比较受试者工作特征曲线(ROC)分析CURB-65、A-DROP、q-SOFA和NLR对患者30 d死亡率的预测效能。结果196例患者30d死亡率为12.7%,CURB-65、A-DROP、q-SOFA和NLR预测死亡率的曲线下面积分别是0.73、0.63、0.69和0.69。结论CURB-65、A-DROP、q-SOFA和NLR均可用于CAP死亡率的预测。其中CURB-65预测效能最高,但易受到医疗条件的限制;qSOFA检测指标要求不高,利于CAP的快速分级。     

经气管镜下APC联合冷冻治疗中央气道病变腔内生长的临床研究

目的探究经气管镜下氩等离子体凝固(APC)联合冷冻治疗中央气道病变腔内生长的临床效果。方法选取2019年1月开始到6月在韶关市第一人民医院诊断及拟行APC联合冷冻治疗的中央气道病变患者30例,采用APC联合冷冻治疗,对比患者治疗前后肺功能,包括:用力肺活量(FVC)、第1秒用力呼气量(FEV 1)、FEV 1/FVC;动脉血气指标,包括:氧分压(PaO 2)、二氧化碳分压(PaCO 2);6分钟步行实验结果(6MWD);气管狭窄程度及气促指数。结果治疗前后患者的FVC无明显差异(P>0.05),治疗后患者的FEV 1%及FEV 1/FVC均较治疗前显著提高(P<0.05),治疗后患者的PaO 2水平较治疗前显著提高且PaCO 2显著下降(P<0.05),治疗后6MWD距离较治疗前显著提高(P<0.05),治疗后患者气管狭窄程度及气促指数分级显著优于治疗前(P<0.05)。结论经气管镜下APC联合冷冻治疗中央气道病变腔内生长的效果显著。     

TRPC1对慢性阻塞性肺疾病慢性气道炎症的影响

目的探讨瞬时感受器电位通道C1(TRPC1)在慢性阻塞性肺疾病(慢阻肺)患者支气管黏膜上皮的表达水平及其与慢性气道炎症之间的关系。方法选取因不明原因肺部结节行纤维支气管镜检查的78例患者,依据慢性阻塞性肺疾病诊治指南分为慢阻肺组(46例)及对照组(32例),其中慢阻肺组再随机分为两个亚组:每日3次规律使用吸入型激素(ICS)布地奈德(ICS组,23例)及安慰剂(非ICS组,23例)治疗。所有受检查者均行支气管肺泡灌洗及支气管镜下刷检。Western Blot及qRT-PCR法检测刷检支气管上皮细胞内TRPC1表达水平,支气管肺泡灌洗液(BALF)行细胞学分类计数。酶联免疫吸附法检测BALF上清液中炎症介质白细胞介素13(IL-13)、成纤维生长因子2(FGF-2)表达水平。结果慢阻肺组支气管上皮细胞内的TRPC1 mRNA、蛋白表达水平明显高于对照组(P<0.05);ICS组的TRPC1表达水平低于非ICS组(P<0.05)。与对照组相比,慢阻肺组BALF中的中性粒细胞、巨噬细胞、淋巴细胞计数及IL-13、FGF-2水平均明显升高(P<0.05)。而ICS组BALF中的中性粒细胞、巨噬细胞、嗜酸性粒细胞、淋巴细胞计数及IL-13、FGF-2表达水平则低于非ICS组(P<0.05)。相关性分析结果显示,TRPC1 mRNA和蛋白表达水平与肺功能第一秒用力呼吸容积(FEV 1)占预计值百分比(FEV 1/Pre%)呈负相关(P<0.01),而与BALF中的中性粒细胞、巨噬细胞、淋巴细胞计数及IL-13、FGF-2水平呈正相关(P<0.05)。结论TRPC1可能通过参与慢阻肺慢性气道炎症过程促进疾病的发生发展,而ICS在一定程度上可干预该作用。     

肺浸润性黏液腺癌1例报告及文献复习

目的：探讨肺浸润性黏液腺癌（IMAs）患者的临床特征、诊断方法和治疗过程,提高临床医生对IMAs的认识。方法：收集1例IMAs患者的一般资料、影像学表现和治疗方案,并进行相关文献复习。结果：患者,女性,42岁,因咳嗽和咳痰入院,CT检查显示双肺斑片状影,因疑似双肺肺炎行抗感染治疗后症状未见好转,经支气管镜肺活检术（TBLB）及胸水脱落细胞检测结果均为炎性表现,后经皮穿刺活检病理回报为IMAs。基因检测结果为KRAS外显子2点突变。行紫杉醇加卡铂并联合应用血管靶向药物贝伐珠单抗化疗后,患者临床症状明显改善,肺部影像学表现明显改变。结论：IMAs是一种特殊病理类型的肺腺癌（ADCs）,该类患者影像学表现多样,基因表达特殊,治疗原则与其他类型ADCs不同。