Objective To investigate the clinical significance of plasma exosomal deleted in lymphocytic leukemia 1(exo-DLEU1)in predicting immunotherapy response of epidermal growth factor receptor(EGFR)/anaplastic lymphoma kinase(ALK)wild type advanced non-small cell lung cancer(NSCLC)patients.Methods The blood samples of 91 patients with EGFR/ALK wild type advanced NSCLC who received immunotherapy in the First People's Hospital of Huzhou,Zhejiang Province from June 2017 to February 2019 were collected and divided into response group(53 cases)and non-response group(38 cases)according to the response after immunotherapy.Plasma exosomes were extracted from the two groups before treatment,and exoDLEU1 expression was detected by real-time fluorescent quantitative PCR.The influencing factors of im-munotherapy response in EGFR/ALK wild type advanced NSCLC patients were analyzed by logistic regression.Receiver operator characteristic(ROC)curve was plotted to evaluate the predictive value of plasma exoDLEU1 level in immunothera-py response.Results Under transmission electron microscopy,the vesicles had circular or elliptical membranes with diame-ters ranging from 30 to 150 nm,and the original concentration was 6.6×1010 particles/ml.Flow cytometry and Western blot showed that the extracts contained exosome markers CD63,CD9,CD81,and heat shock protein 70.Real-time fluorescence quantitative PCR results showed that the plasma exoDLEU1 level in the response group before treatment was lower than that in the non-response group,and the difference was statisti-cally significant(P＜0.05).Analysis of influencing factors showed that plasma exoDLEU1 expression level was an indepen-dent influencing factor for immunotherapy non-response in EGFR/ALK wild type advanced NSCLC patients(OR=6.119,95%CI:2.768-13.526,P＜0.05).ROC curve analysis showed that plasma exoDLEU1 expression level predicted im-munotherapy response of EGFR/ALK wild type advanced NSCLC patients with an area under the curve of 0.900(P＜0.05).Conclusion The plasma exoDLEU1 level of EGFR/ALK wild type advanced NSCLC patients before immunotherapy can be used as an indicator to predict their treatment response,and help to screen out patients who are more likely to benefit from immunotherapy in the clinic.

目的 探讨血清外泌体长链非编码RNA(LncRNA)在非小细胞肺癌(NSCLC)早期诊断中的价值.方法回顾性选取湖州市第一人民医院2017年 1月至2020年3月检测的206份NSCLC和200份健康对照人群的血清样本,随机分为训练集(50例NSCLC和50例健康对照)和验证集(156例NSCLC和150例健康对照).从NSCLC和健康对照的血清样本中分离外泌体,进行外泌体RNA深度测序以检测差异表达的外泌体LncRNA,然后利用实时荧光定量聚合酶链式反应(qRT-PCR)分析在训练集和验证集队列中表达失调最显著的外泌体LncRNA PCAT1(exoPCAT1),采用受试者工作特征(ROC)曲线分析检测外泌体生物标志物的诊断能力.结果 共发现781个外泌体LncRNA在NSCLC和健康对照组血清样本之间差异表达(变化倍数＞1.5,P＜0.05).在训练集中,利用qRT-PCR证实exoPCAT1在NSCLC血清样本中较健康对照组过表达[2.24(1.45,3.44)vs 0.92(0.57,1.30),Z=5.667,P＜0.05].在验证集中,NSCLC组exoPCAT1 相对表达量亦高于健康对照组[2.09(1.32,3.18)vs0.84(0.56,1.41),Z=10.112,P＜0.05].exoPCAT1 诊断 NSCLC 的曲线下面积(AUC)为 0.838(95％CI0.795～0.880).对于早期(Ⅰ～Ⅱ期)或者血清癌胚抗原正常(＜5 ng/ml)的NSCLC患者,exoPCAT1也有较高的诊断价值,AUC分别为0.789(95％CI 0.737～0.841)和 0.728(95％CI 0.661～0.795).另外,血清 exoPCAT1 表达与疾病进展有关,高表达(＞2.09)者TNM Ⅲ～Ⅳ期、T1～T2分期及淋巴结转移阳性的比例更高(均P＜0.05).结论 血清外泌体LncRNA PCAT1可作为一种新的"液体活检"生物标志物,并可用于早期NSCLC的诊断.

目的 分析湖州市近年来肠道门诊腹泻患者病原菌的检出情况,给当地感染性腹泻疾病防控提供科学依据.方法 收集该院2015—2019年就诊的腹泻患者3499例的粪便标本,按照《浙江省食源性致病菌监测工作手册》进行细菌培养鉴定.结果 共分离出阳性菌株769株(22.0％),检出菌株种类有致泻性大肠埃希菌(9.5％)、副溶血性弧菌(5.0％)、嗜水/豚鼠气单胞菌(3.4％)、沙门菌(3.1％)、非O1/非O139霍乱弧菌(0.5％)、河流弧菌(0.5％).2017—2019年病原菌检出率逐年下降.腹泻主要发生在21～30岁(24.3％)及31～40岁(17.0％),而致病菌检出率以71～80岁最高(35.0％).结论 湖州地区腹泻病原菌种类多,居首的是致泻性大肠埃希菌,患者以青壮年为主,致病菌检出率以老年患者最多,防控措施需要根据本地区的流行特点实施.

目的 了解湖州地区食源性腹泻病原谱的分布特征,可疑中毒食物范围和感染的季节分布为临床诊断和制定防控措施提供依据.方法 通过对湖州地区2016年食源性腹泻患者的粪便进行细菌病原学检测,并对检测结果进行分析.结果 510例食源性腹泻患者的粪便共分离得到病原菌110株,检出率21.57％,检出的病原菌包括致泻性大肠杆菌60株11.76％(60/510)、副溶血性弧菌41株8.04％(41/510)、沙门菌5株0.78％(5/510)、气单胞菌3株0.59％(3/510)、其它肠道病原菌2株0.40％(2/510),志贺氏菌未检出.510例食源性腹泻患者中主要可疑食物为肉类与肉制品24.2％,果蔬类占18.1％,混合多种食品占17.1％,水产品类占13.9％.结论 湖州地区食源性腹泻感染高发期为8月份,病原菌前三位是致泻性大肠杆菌,副溶血性弧菌,沙门菌,同时肉类和肉制品、果蔬类、混合多种食品和水产品是可能的污染食品.

目的 分析食源性腹泻患者副溶血弧菌(VP)季节分布及感染患者年龄特征、毒力基因、血清学分型.方法 对2014年1月至2016年12月急性腹泻患者粪便分离的125株副溶血弧菌进行常规培养、血清分型及PCR毒力基因检测.结果 副溶血弧菌感染发生有明显的季节性,夏秋季高发,尤以6～10月份为高峰期.感染患者中21～30岁占37.6％,31～40岁占24.8％.分离得到17个血清型,其中O3:K6占59.2％.经单重PCR进行毒力基因检测显示:tdh+/trh-分离株所占比例为88.0％,tdh-/trh-分离株为6.40％,检测到2株携带(tdh+/trh+)基因.结论 湖州地区副溶血性弧菌感染发生呈明显的季节分布特征,且感染的患者以青壮年居多,优势血清型为O3:K6;多数菌株携带tdh+/trh-毒力基因.