目的 探讨穿透素3(PTX3)、可溶性髓系细胞受体-1(sTREM-1)、细胞因子信号传导抑制因子3(SOCS3)在细菌性脓毒症患者血清中的水平变化及诊断价值.方法 本研究选取2019年1月至2022年12月在本院治疗的细菌性脓毒症患者94例作为观察组,同期选取非感染性全身性炎症性反应综合征(SIRS)患者80例及健康体检志愿者100例分别纳入对照组和健康组.检测血清PTX3、sTREM-1、SOCS3水平;受试者工作特征曲线(ROC)评价血清PTX3、sTREM-1、SOCS3水平对细菌性脓毒症患者的诊断价值;采用多因素Logistic回归分析法分析细菌性脓毒症的影响因素.结果 观察组患者血清PTX3、sTREM-1、SOCS3表达水平分别为(104.39±23.93)ng/mL、(136.17±41.46)ng/mL、(346.59±31.12)μg/L,高于对照组患者血清PTX3、sTREM-1、SOCS3表达水平(53.93±12.19)ng/mL、(69.17±14.46)ng/mL、(271.19±26.02)μg/L及健康组患者血清PTX3、sTREM-1、SOCS3表达水平(1.17±0.33)ng/mL、(10.15±3.17)ng/mL、(163.32±17.33)g/L,三组间比较差异均有统计学意义(F=5.928,588.015,11.785,P＜0.05);ROC结果显示,血清PTX3、sTREM-1、SOCS3水平预测细菌性脓毒症患者的曲线下面积(AUC)分别为0.867、0.878、0.893,对应的敏感度分别为90.43％、78.72％、81.91％,特异度分别为86.25％、88.75％、90.00％,三者联合预测细菌性脓毒症患者的AUC为0.957,敏感度为92.55％,特异度为85.00％;Logistic回归分析显示PTX3、sTREM-1、SOCS3水平升高均是细菌性脓毒症的危险因素.结论 PTX3、SOCS3、sTREM-1在细菌性脓毒症患者血清中均呈明显高表达,三者联合检测有助于诊断疾病的发生.
目的 检测细菌性脑膜炎(BM)患者脑脊液、血清中lncRNA NEAT1水平变化情况,并分析其与患者病情严重程度以及血脑屏障破坏程度的相关性.方法 收集2015年2月-2021年12月间本院收治的BM患者66例作为BM组,另选取同期不具备神经系统感染性疾病但具有腰穿指征的同期住院患者40例作为对照组.根据中枢神经系统功能障碍程度将患者分为普通BM组与重症BM组,收集患者入院48 h内脑脊液与血清标本,qRT-PCR法检测脑脊液与血清标本中lncRNA NEAT1水平,比较对照组、BM组间以及普通BM组与重症BM组间脑脊液与血清标本中lncRNA NEAT1水平;根据白蛋白指数对BM患者血脑屏障破坏程度进行分级,比较普通BM组与重症BM组血脑屏障破坏程度;Pearson法分析脑脊液lncRNA NEAT1、血清lncRNA NEAT1、白蛋白指数间相关性.结果 与对照组相比,BM 组患者脑脊液(3.28±0.51 vs 1.01±0.18)、血清 lncRNA NEAT1 水平(2.51±0.42 vs 1.01±0.16)均升高(P＜0.05),且BM组患者血清与脑脊液中lncRNA NEAT1水平比较差异有统计学意义(P＜0.05);与普通BM患者相比,重症 BM 患者脑脊液 lncRNA NEAT1(3.62+0.53 vs 2.82±0.46)、血清 lncRNA NEAT1(2.70±0.46 vs 2.25±0.37)水平升高(P＜0.05),且普通BM、重症BM患者血清与脑脊液中lncRNA NEAT1水平比较差异有统计学意义(P＜0.05);普通BM患者、重症BM患者血脑屏障损伤程度之间比较差异有统计学意义(P＜0.05);BM患者脑脊液、血清lncRNA NEAT1均与白蛋白指数呈正相关(P＜0.05),BM患者脑脊液lncRNA NEAT1与血清lncRNA 结论 BM患者脑脊液、血清中lncRNA NEAT1高表达,其水平与患者严重程度、血脑屏障损伤程度密切相关,为BM患者的诊疗提供了靶点.
目的 探讨天麻素(Gas)对呼吸道合胞病毒(RSV)感染肺炎大鼠肺组织的保护机制及对Th17/Treg细胞平衡的影响.方法 将Wistar雄性大鼠60只分为对照组(C组)、模型组(M组)Gas低剂量组(Gas-L,5 mg/kg)、Gas中剂量组(Gas-M,10 mg/kg)、Gas高剂量组(Gas-H,20 mg/kg)、阳性组(0.15 g/kg利巴韦林),每组10只.药物干预结束后,称定大鼠体质量和肺组织干湿重,计算肺指数和肺组织W/D值;取肺组织作HE染色检查并进行损伤评分;qRT-PCR检测肺组织RSV病毒载量;流式细胞术检测脾组织Th17、Treg细胞比例;ELISA法检测血清TNF-α、IL-1β、IL-6、IL-17A、TGF-β含量;Western blot检测肺组织Notch/NF-KB通路蛋白表达.结果 与C组相比,M组肺组织出现中性粒细胞浸润,肺泡破裂连片等病变,肺指数、W/D值、肺损伤评分、RSV病毒载量、Th17细胞比例、TNF-α、IL-1β、IL-6、IL-17水平、Notch-1、Hes-1、-NF-KB p65/NF-KB p65蛋白均显著升高(均P＜0.05),Treg细胞比例和TGF-β水平均显著下降(均P＜0.05).Gas-M、H组和阳性组肺损伤程度显著改善,肺指数、W/D值、肺损伤评分、RSV病毒载量,Th17细胞比例,TNF-α、IL-1β、IL-6、IL-17 及 Notch-1、Hes-1、p-NF-κB p65/NF-κB p65 蛋白水平均显著下降(均 P＜0.05),Treg细胞比例和TGF-β水平显著上升(均P＜0.05).结论 Gas具有减轻RSV感染肺炎大鼠的肺组织炎症损伤和维持Th17/Treg细胞平衡的作用,可能通过抑制Notch/NF-κB信号通路实现.
目的 克隆羊口疮病毒020基因,构建其重组真核表达质粒并转染HeLa细胞,分析020基因编码E3L蛋白的亚细胞定位,分析该蛋白生物信息学特征.方法 提取羊口疮病毒基因组,PCR扩增020基因并构建重组真核表达质粒,转染HeLa细胞,采用Western blot检测其在该细胞中的表达,免疫荧光法检测该蛋白的亚细胞定位.利用DNAstar软件分析不同毒株ORFV E3L氨基酸序列特点及其遗传演化关系;利用SOMPA软件分析其二级结构;利用SignalP4.0软件预测信号肽;利用TMHMM2.0软件预测跨膜结构;利用NetPhos 3.1预测磷酸化位点;利用NetNGlyc-1.0预测糖基化位点;应用IDEB和SYFPEITHI预测ORFV E3L蛋白的抗原表位.结果 成功克隆得到ORFV 020基因,全长549 bp,编码183个氨基酸;亚细胞定位显示该蛋白定位于细胞核和细胞质.生物信息学分析ORFV E3L蛋白α螺旋约占40.44％、β折叠约占5.46％、延伸链约占16.94％、无规则卷曲约占37.16％;该蛋白无信号肽和跨膜结构,可能存在15个磷酸化位点,7个N-糖基化位点,5个B细胞线性表位,2个CTL表位,3个Th细胞表位,3个B细胞和CTL联合表位,3个B细胞和Th细胞联合表位.结论 ORFV E3L蛋白定位于细胞核和细胞质,含有多个抗原表位,可能具有免疫原性.该蛋白高度保守,具有成为优势保护性抗原的潜力.为进一步揭示ORFV E3L蛋白的功能奠定基础,为ORFV诊断方法的建立及疫苗的研制提供了理论依据.
目的 探究干扰Wnt5a对卡介苗(BCG)感染的小鼠肺上皮细胞(TC-1)炎性反应的调控作用,并阐明其作用机制.方法 慢病毒干扰Wnt5a处理TC-1细胞系及其阴性对照细胞系后BCG感染24h.设置4个实验组:NC组、NC+BCG 组、Si-3 组和 Si-3+BCG 组.通过 Western blot 检测 Toll 样信号通路 TLR2、TLR4、Myd88、p-NF-κB 和 NF-κB炎症相关蛋白表达水平,免疫荧光法检测各实验组p-NF-κB表达情况,qRT-PCR检测Toll样信号通路TLR2、TLR4、Myd88、p-NF-KB和NF-κB等基因mRNA表达水平,ELISA检测细胞培养上清促炎因子TNF-α、IL-6和IL-10含量.结果 与对照组比较,BCG组炎性相关因子TLR2、TLR4、Myd88、NF-KB、p-NF-κB的蛋白表达水平(均P＜0.01)和mRNA表达水平均显著上调(均P＜0.05),p-NF-κB蛋白荧光强度较对照组显著增加,促炎因子TNF-α和IL-6表达水平升高(均P＜0.01),抑炎因子IL-10表达水平降低.与BCG组相比,Si-3+BCG组上述蛋白表达水平(均P＜0.05)和mRNA表达水平均(均P＜0.01)显著下调,p-NF-κB蛋白荧光强度较BCG组均显著下调,促炎因子TNF-α和IL-6(均P＜0.05)表达水平显著降低,抑炎因子IL-10表达水平显著升高(P＜0.01).结论 干扰Wnt5a可调控卡介苗(BCG)感染的小鼠肺上皮细胞(TC-1)的炎性反应,其调控作用机制是通过Toll样信号通路依赖Myd88途径抑制BCG诱导的TC-1炎性反应.
Malaria is transmitted by the bite of Anopheles mosquitoes with the parasite, mainly prevalent in tropical and subtropical regions. Malaria is prevalent in Bangladesh. This review reviews the recent progress on malaria epidemic characteristics in Bangladesh to provide theoretical.
Objective Analysis of the antigenic epitope of the outer membrane protein H.8,expression and purification of the protein using a prokaryotic expression system. Methods The target protein was analyzed by bioinformatics technology to obtain its corresponding physical, chemical properties and prediction of B cell and T cell epitopes.The out membrane protein H.8 gene was cloned into pET32a（+）,and its prokaryotic expression system was constructed using the E.coli expression hosts, Rossetta（DE3）.The fusion protein was detected by SDS-PAGE,and isolated and purified by nickel column affinity chromatography to verify its immunoreactivity by Western blot assay. Results Peptides19-57,63-70,73-83,90-100,108-135,140-150 and 153-163 of the out membrane protein H.8 were the dominantB cell epitopes, while peptides 125-139 and 132-146 were the dominantT cell epitopes.The recombinant protein was significantly expressed in the supernatant at IPTG concentration of 0.5 mmol/L after transformation of DE3 with the constructed pET32a（+） recombinant plasmid containing the outer membrane protein H.8 gene.The results of Western blot showed that the recombinant protein can recognize by His-tag antibody andpolyclonal antibody of N.gonorrhoeae respectively. Conclusion Bioinformatics methods predicted that N. gonorrhoeae outer membrane protein H.8 contains dominant B-cell epitopes and dominant T-cell epitopes, and that the outer membrane protein H.8 expressed in prokaryotic cellshas immunoreactive, which providing an experimental basis for the study of N. gonorrhoeae vaccines and its pathogenic mechanisms.
目的 分析开封地区细菌性痢疾患儿的临床表现、流行病学特征、病原菌及耐药率.方法 选取2019-2021年开封市儿童医院小儿内科就诊的腹泻患儿,其中232例被确诊为细菌性痢疾.制定病历调查表,采用一对一方式,收集患儿资料(包括基础资料、临床表现、病原菌及耐药性),分析细菌性痢疾患儿的临床表现及流行病学特点.参与本次研究的所有患儿,进行抗菌治疗前,采集患儿新鲜粪便标本.接种于培养基进行培养,挑取菌落进行病原菌鉴定及血清凝集试验.采用K-B纸片扩散法,进行志贺菌属对氨苄西林、氨苄西林/舒巴坦、头孢他啶、头孢曲松、头孢吡肟、亚胺培南、美罗培南、环丙沙星、左氧氟沙星及复方新诺明的药敏试验.结果 232例细菌性痢疾患儿的临床表现中,腹痛、呕吐、阵发性哭闹、每日腹泻次数＞10次、每日腹泻次数≤10次、低热、中热、高热、黄色稀水样便、黄色糊状便、黏液脓血便的发生率分别为 100％、70.26％、75.86％、33.19％、66.81％、42.24％、31.03％、26.72％、27.59％、11.21％和61.21％.其中腹痛、阵发性哭闹、呕吐,居临床表现前3位.按照患儿发病年龄分组,＜1岁组12例,1～岁组42例,3～岁组60例,5～岁组37例,7～岁组33例,9～岁组28例,11～岁组20例.232例细菌性痢疾患儿,2019年85例,2020年77例,2021年70例.每年1月份的发病率最低,分别为1.18％、1.30％和1.43％.发病率逐月上升,每年7月份为高发期,分别为20.00％、19.48％和21.43％.8月份开始发病率逐月下降,12月份的分别为2.35％、2.60％和2.86％.共检出病原菌216株,阳性率93.10％.2019年检出85株,2020年70株,2021年61株,阳性率分别为100％、90.91％和87.14％,患儿粪便标本病原菌阳性率呈现逐年下降趋势.检出福氏志贺菌137株,各年度分别检出61、43和33株,检出率逐年下降.检出宋内志贺菌60株,各年度分别检出19、20和21株,检出率逐年上升.检出鲍氏志贺菌14株,各年度分别检出4、5和5株,检出率逐年上升.检出痢疾志贺菌5株,各年度分别检出1、2和2株,检出率逐年上升.216株志贺菌属对氨苄西林、氨苄西林/舒巴坦、头孢他啶、头孢曲松、头孢吡肟、亚胺培南、美罗培南、环丙沙星、左氧氟沙星及复方新诺明的药敏试验显示,各年度均未产生亚胺培南、美罗培南的耐药株.对氨苄西林、氨苄西林/舒巴坦、复方新诺明的耐药率较高,各年度均高于50％,并呈现逐年升高的趋势.对头孢他啶、头孢吡肟的耐药率较低,各年度均低于50％.头孢曲松的耐药率,各年度分别为40.00％、45.71％和54.10％,呈现快速上升的变化趋势.结论 开封地区小儿细菌性痢疾,发病患儿主要集中于3-5岁,高发于每年7月份.以福氏志贺菌为主要病原菌,对多种抗菌药物的耐药性逐年上升,药物敏感性发生变化,临床上应重点关注本地区志贺菌属亚型及耐药情况.
目的 调查老挝琅勃拉邦省蚊虫种类组成及重要生态习性,为当地制定有效的媒介控制策略及措施提供参考.方法 2022年11月,在老挝北部琅勃拉邦省采用诱蚊灯通宵捕蚊法、人工帐诱捕法和人牛房全捕法采集成蚊,采用勺舀法捞捕居民周边小积水伊蚊四龄幼虫,采集的成蚊和伊蚊幼虫通过形态学方法进行种类鉴定并登记.结果 共捕获成蚊2亚科6属29种2149只,其中三带喙库蚊和乌头按蚊属当地优势种,分别占捕获总数的19.48％(419/2149)和16.54％(345/2149);共捕捞伊蚊幼虫1属2种14条,其中埃及伊蚊和白纹伊蚊占比分别为85.71％(12/14)和14.29(2/14).微小按蚊和多斑按蚊通宵均有叮咬活动,叮咬高峰分别主要出现在子夜(00:00-01:00)和黄昏(20:00-21:00).结论 老挝琅勃拉邦省蚊虫种类丰富,三带喙库蚊为优势蚊种,同时存在具有夜间叮咬活动的微小按蚊,以及埃及伊蚊和白纹伊蚊,提示当地仍然存在疟疾、登革热和流行性乙型脑炎疫情风险,建议加强对上述疾病的监测.
The m6A methylation of RNA in eukaryotic cells affects gene function by regulating the splicing, localization, stabilization and translation of RNA,and methyltransferase, demethylase and reading proteins are involved in the methylation modification process. In recent years, it has been found that in the battle between virus and host, and host m6A modification system might affect viral replication by regulating viral RNA. In the same time, viral proteins also affect the overall m6A modification extent by influencing methylation modification-related factors to change the level of cytokines, interfere with the proliferation and the activity of immune cells, and reduce stress response, ultimately toensure that the virus replicates successfully. This article summarizes the roles of m6A modification during the infection and replication of three types of RNA viruses, including influenza A virus（IAV）,human immunodeficiency virus type I（HIV-I）,and novel coronavirus（SARS-CoV-2）,as well as three types of DNA viruses: hepatitis B virus（HBV）,Epstein Barr virus（EBV）,and adenovirus（AdV）. These studies contribute to explore the inhibitors targeting m6A regulatory factors and will provide a theoretical clue toward the development of novel antiviral drugs.
目的 分析2型糖尿病合并肺部感染患者的病原菌分布及肺部CT特征.方法 选取173例于新疆医科大学第五附属医院进行治疗的2型糖尿病合并肺部感染患者为研究对象.于患者接受治疗前,采集其痰液、支气管肺泡灌洗液后采用全自动细菌鉴定分析仪进行病原菌鉴定.所有患者在治疗前、后均进行肺部CT检查,观察不同肺部感染类型患者的CT影像特点.患者经治疗后,观察记录患者血糖控制情况与疗效,分析患者血糖控制情况与肺部感染治疗效果的相关性.结果 共检出病原菌173株.革兰阳性菌42株(24.28％),主要为肺炎链球菌(15.61％)、金黄色葡萄球菌(5.78％).革兰阴性菌112株(64.74％),主要为肺炎克雷伯菌(15.03％)、鲍曼不动杆菌(12.72％).经CT检查,肺炎154例,以小叶肺炎(45.09％)为主,肺结核患者10例,真菌感染9例.不同肺部疾病患者的CT影像特点具有差异化.大叶肺炎患者CT影像显示大片状实变阴影沿肺段分布;小叶肺炎患者CT影像显示,点、斑片状沿支气管血管分布,肺结核患者CT影像显示均为活动性肺结核,15个肺段产生病灶,全部伴有肺门纵膈淋巴结肿大;肺曲菌病患者CT影像空洞内可见可移动圆形结节影;肺念珠病患者CT影像显示大块成片状实变影.肺部感染患者抗菌药物使用情况,主要为单联使用药物方式(86.71％),以头孢菌素、第三代喹诺酮类为主.11.56％采用两联使用抗菌药物,头孢菌素+喹诺酮类使用频次最高.173例2型糖尿病合并肺部感染患者经过治疗后,136例患者血糖控制良好,患者痊愈、好转率为86.76％,显著高于血糖控制不良患者组,差异有统计学意义(x2=14.049,P＜0.05).结论 2型糖尿病合并肺部感染患者病原菌主要以肺炎克雷伯菌、鲍曼不动杆菌、肺炎链球菌为主,肺部感染类型多小叶肺炎.临床治疗一般以以头孢菌素、第三代喹诺酮类为主,控制血糖有助于肺部感染的治疗.
目的 分析儿童急性化脓性阑尾炎术后感染病原菌分布及不同用药方案疗效.方法 选取2019年1月1日～2022年7月30日本院收治的569例急性化脓性阑尾炎患儿作为研究对象.设计并制作临床资料收集表,收集患儿基本情况、临床症状、病原学检查结果等.所有患儿进行阑尾炎切除手术,术中吸取患者阑尾腔分泌物.诊断为术后切口感染的患儿,采集其切口分泌物,分别接种于不同培养基内进行培养,提纯分离后,使用全自动微生物分析仪进行病原菌鉴定.采用Pearson分析术后切口感染患者的阑尾腔分泌物与切口分泌物病原菌之间的相关性.将参与本次研究的569例急性化脓性阑尾炎患儿随机分为观察组和对照组,两组患儿均进行腹腔镜下切除术.对照组患儿术后给予甲硝唑注射液,以静脉注射方式连续治疗3 d.观察组患儿在对照组患儿治疗基础上联合使用盐酸头孢吡肟注射液,以静脉注射方式连续治疗3 d.对比两组患儿术前及术后血清炎症因子水平测得值、术后切口感染率.结果 569例急性化脓性阑尾炎患儿中,发生术后切口感染68例,感染率为11.95％.67例患儿出现明显腹痛,其中30例为阵发性腹痛.对68例患儿采集外周血进行血液分析,66例患儿白细胞计数＞10×109/L,65例患儿C反应蛋白＞10 mg/L.采集68例急性化脓性阑尾炎术后切口感染患儿的阑尾腔分泌物进行培养分离,共分离到病原菌59株.89.83％为革兰阴性菌,以大肠埃希菌、肺炎克雷伯菌为主.采集68例术后切口感染患儿的切口分泌物进行培养分离,共分离到病原菌75株,61例患儿为单一病原菌感染.84.00％为革兰阴性菌,以大肠埃希菌、肺炎克雷伯菌为主,16.00％为革兰阳性菌,主要为金黄色葡萄球菌.对68例急性化脓性阑尾炎术后切口感染患儿的阑尾腔分泌物与切口分泌物病原菌两变量高度相关(r=0.997,P＜0.05).对照组患儿术后切口感染率为15.85％,观察组患儿术后切口感染率为8.07％,两组患儿术后发生切口感染率对比差异具有统计学意义.两组患儿手术前血清中白细胞介素-8(IL-8)、肿瘤坏死因子-a(TNF-a)水平对比差异无统计学意义(P＞0.05),观察组患儿术后血清中IL-8、TNF-a水平较术前明显下降,对比差异有统计学意义(P＜0.05).结论 急性化脓性阑尾炎患儿术后切口感染病原菌主要以革兰阴性菌为主,术后切口感染患者的阑尾腔分泌物与切口分泌物病原菌之间的相关性较强.术后患儿采用头孢吡肟与甲硝唑联合用药方案,可以有效预防术后发生切口感染,对患儿血清炎症因子水平具有显著影响.
Objective In this study, we applied reverse genetic techniques to construct a chimeric influenza vaccine expressing the HA protein of seasonal influenza virus H1N1 using the cold-adapted attenuated influenza B virus strain as a backbone. Methods The HA fragment ectodomain region of B/Vienna/1/99 of influenza B virus was replaced with the HA protein of H1N1 influenza virus（A/Victoria/2570/2019）,and the recombinant plasmid was cotransfected with the other seven plasmids of the backbone strain in 293T cells at 33 ℃ under 5% CO 2 for 72 hours to rescue the H1N1 chimeric vaccine strain. The recombinant viral strain was subjected to hemagglutination assay identification, RT-PCR identification, electron microscopy identification, one-step growth curve. We evaluated the safety of this H1N1 chimeric influenza vaccine in mice model. Results The H1N1 chimeric influenza virus strain was successfully rescued and named rA/B-H1-Vic. The sequence was sequenced correctly and identified as expected, and typical particles of influenza virus were observed by electron microscopy. The H1N1 chimeric influenza virus strain had a hemagglutination titersof about 2~5. Also, we detected the viral titer of 10 4.5 TCID 50 /mL on MDCK cells and 10 8.36 EID 50 /mL in chicken embryo eggs.Mice were inoculated intranasally with 10~6EID 50 and 10~5EID 50 in a volume of 50 μL,respectively, and the infective mice showed no significant weight loss compared to control mice, the survival rate of infective mice was 100%. We detected a low level of viral replication only in nasal turbinate and lungs-on day 3 post-infection. The other tissues, including hearts, levers, spleens, kidney, intestines, and brains, were found no virus replication. Conclusion We rescued the non-virulent H1N1 chimeric vaccine strain successfully and it may provide a new strategy for the development of novel influenza vaccines.
Objective To explore the basic biochemical properties of the protein encoded by Rv1511 gene of Mycobacterium tuberculosis, and provide experimental data for the recombinant expression of the protein in Escherichia coli,evaluated the feasibility of it as a marker for tuberculosis detection. Methods The Rv1511 gene and amino acid sequence were obtained by bioinformatics method, it is basic biochemical properties were predicted. The recombinant expression vector pET28a（+）-Rv1511 with Escherichia coli as host was constructed, the high purity recombinant protein was obtained by nickel metal chelate chromatography and verified by Western blot. Results The protein encoded by Rv1511 gene consists of 340 amino acids. The protein is GDP-d-mannose dehydrase, which is related to the pathway of glucose metabolism and may be involved in the permeability of cell wall control substances. The protein is a hydrophilic intracellular protein without transmembrane domain, with theoretical molecular weight of 38.34241×10~3and isoelectric point of 6.08. It has 36 phosphorylation sites and 1 N-glycosylation site. BepiPred-2.0 predicted that the protein contained 4 B cell epitopes, SYFPEITHI and Rankpep predicted 8 CTL cell epitopes and Th cell epitopes, NetMHCIIpa4.0 predicted T cell epitopes showed that the protein had multiple MHCI and MHCII binding sites. The recombinant plasmid was constructed and expressed in background after transfection of E.coli BL21（DE3）. The expressed product was insoluble inclusion body. The target protein with a purity of 92.6% was obtained by nickel metal chelation purification after 8mol/L urea denaturation. SDS-PAGE and mouse anti-histidine protein antibody Western blot confirmed that the recombinant protein was successfully expressed in Escherichia coli. Conclusion Rv1511 gene and coding protein are deleted in BCG,which is expected to be a potential marker for clinical detection of tuberculosis. The protein has multiple modification sites and antigenic epitopes, which may be related to the stress resistance of Mycobacterium tuberculosis under hypoxia and nutritional deficiency, and is a potential target for anti-tuberculosis drugs. The recombinant protein of Rv1511 gene has reactivity, which can provide experimental support, further evaluation of its feasibility as a marker for tuberculosis detection.
Objective To explore the relationship between CD64 infection index, interleukin-6（IL-6） and the prognosis of patients with multidrug resistant Acinetobacter baumannii bloodstream infection. Methods A total of 102 patients with multidrug resistant A. baumannii bloodstream infection admitted to our hospital from October 2017 to October 2022 were selected as the study group, according to the prognosis of patients, they were divided into poor prognosis group（n=57） and good prognosis group（n=45）. In addition, 98 patients with pulmonary infection and mechanical ventilation were regarded as the control group. Clinical data were collected and Cox multifactor analysis was applied to analyze the factors affecting the prognosis of patients with multidrug resistant A. baumannii bloodstream infection; ROC curve was applied to analyze the predictive value of CD64 infection index and IL-6 level in peripheral blood on the prognosis of patients with multidrug resistant A. baumannii bloodstream infection. Results Compared with the control group, the peripheral blood CD64 infection index[（6.71±1.06）,（2.85±0.57）] and IL-6 levels[（206.91±22.92）,（137.52±15.34）ng/L] in the study group were obviously higher（P＜0.05）. Compared with the good prognosis group, the peripheral blood CD64 infection index[（8.25±1.39）,（4.77±0.84）] and IL-6 levels[（245.86±29.51）,（157.57±16.92）ng/L] in the poor prognosis group were obviously higher（P＜0.05）. The prognosis of patients with multidrug resistant A. baumannii bloodstream infection was independently affected by the complicated immunosuppressive status, antibacterial drug use type ≥3,Acute Physiological And Chronic Health Evaluation II（APACHE II） score, mechanical ventilation time, CD64 infection index and IL-6（P＜0.05）. ROC curve analysis showed that the AUC of CD64 infection index alone predicting the prognosis of patients with multidrug resistant A. baumannii bloodstream infection was 0.869,the best cutoff value was 5.66,and the sensitivity and specificity were 70.27% and 86.99%,respectively; the AUC of IL-6 alone in predicting the prognosis of patients with multidrug resistant A. baumannii bloodstream infection was 0.836,the best cutoff value was 239.15 ng/L,and its sensitivity and specificity were 67.57% and 86.30%,respectively, the AUC predicted by the combination of the two was 0.926,obviously higher than that predicted by the CD64 infection index alone（Z=3.849,P=0.000） and the level of IL-6 alone（Z=2.233,P=0.026）. Conclusion The CD64 infection index and IL-6 level in peripheral blood are obviously increased in patients with multidrug resistant A. baumannii bloodstream infection, the combined detection of the two has high predictive value for the prognosis of patients with multidrug resistant A. baumannii bloodstream infection.
Objective To investigate the infection status of human soil-transmitted helminth（STH） in China-Vietnam border areas, providing local scientific basis for formulating prevention and control measures. Methods Jinping County was selected as the site of investigation by adopting cluster random sampling method and followed by selecting one administrative village from each area. Collect the feces of the subjects and the eggs of STH in feces were detected by the Kato-Katz method. The culture method of test tube filter paper was used to identify hookworm species. The eggs of Enterobius vermicularis（E. vermicularis） were detected by the cellophane tape method in children at aged of 3-9 years-old. Results The stool samples from 1350 residents were examined and the STH infection rate was 14.15%（191/1350）. The infection rates of Ascaris lumbricoides（A.lumbricoides）,whipworm, hookworm and Enterobius vermicularis（E. vermicularis） were 0.30%（4/1350）,3.70%（50/1350）,13.11%（177/1350）and 0.44%（6/1350） respectively. The eggs of E. vermicularis were detected by the cellophane tape method in children aged 3-9 years, a total of 154 children were examined and 5 were infected. A total of 52 cases of hookworm infection were identified by test tube filter method, all were infected with. Minor infections with A.lumbricoides,whipworm and hookworm accounted for 75.00%（3/4）,88.00%（44/50） and 85.88%（152/177）respectively.The highest infection rate of STH was found in the Ding qing village of Zhe mi Lahu nationality township（22.63%） among the five villages investigated. The infection rate of STH was statistically significant among different village（χ~2=33.28,P＜0.05）.The infection rate of STH was 12.26%（82/669）in male and 16.01%（109/681） in female, with significant difference（χ~2=3.90,P＜0.05）. The STH infections were found in all age groups with the highest infection rate in the age group of 60 and above years（22.12%）. People in Lahu ethnic group owned higher STH infection rate of 22.71%than people with other ethnic groups. Farmers possessed the highest infection rate of 15.25% compared with people with other occupations. In terms of education, the STH infection rate was highest among primary school residents（15.84%）. The infection rate of STH was statistically significant among different age groups, nationalities, and educational levels（χ~2=19.43、22.97、9.86,P＜0.05）. Conclusion The infection rate of STH is still at a high level in Jinping county of China-Vietnam border areas and the results were showed that local department should be strengthen the prevention and treatment of soil-transmitted helminth for farmers, the elderly and ethnic groups.
目的 研究华支睾吸虫来源的分子伴侣(molecular chaperones HscB,CsHscB)对氧化偶氮甲烷(azoxymethane,AOM)联合葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导的小鼠炎症性结肠癌的治疗效果及潜在的分子机制.方法 C57BL/6 小鼠随机分为正常对照组(NC,n=10),rCsHscB组(n=10),模型组(AOM+DSS,n=15),干预组(AOM+DSS+rCsHscB,n=15).模型组和干预组按8 mg/kg体重的剂量经腹腔注射AOM,1 次/d,连续7 d.然后给予7 d 2％DSS饮水和14 d正常饮水,共4个循环.在给予DSS饮水的第4 d、7 d,rCsHscB组和干预组小鼠按照1.25 mg/kg体重的剂量经腹腔注射rCsHscB蛋白.记录小鼠体重和生存状况,第84 d剖杀小鼠,测量结肠长度,记录结肠肿瘤个数,HE和Masson染色观察结肠病变,ELISA检测结肠组织中IL-6、MCP-1 和IL-10 含量,Western blot检测结肠组织中α-SMA、MMP2、Occludin表达水平和ERK1/2、JNK、P38、AKT、STAT3 的磷酸化及非磷酸化水平.结果 与正常对照组相比,模型组71.4％的小鼠结肠中出现肿瘤,小鼠体重和结肠长度显著降低缩短(均P＜0.05),结肠组织中有大量炎症细胞浸润和纤维化形成.与模型组相比,干预组小鼠的生存率显著升高(P＜0.01),体重、结肠长度和炎症纤维化均有所改善,仅有10％小鼠出现结肠肿瘤(P＜0.01).与模型组相比,干预组小鼠结肠组织中IL-6 和MCP-1 显著降低(均P＜0.05),IL-10 显著升高(P＜0.0501),α-SMA 和MMP2 蛋白表达水平降低(均P＜0.05),Occludin蛋白表达水平升高(P＜0.05),ERK1/2、JNK、P38、AKT和STAT3蛋白的磷酸化和非磷酸化水平均显著降低(均P＜0.05).结论 rCsHscB可能通过抑制MAPK和AKT/STAT3 信号通路的活化改善AO M/DSS诱导的小鼠炎症性结肠癌.
目的 分析血液病患者并发血流感染的病原菌分布特点及危险因素.方法 选取2021年1月～2021年12月于本院血液内科发生血流感染的血液病患者150例,同时随时选取同期60例未发生血流感染的血液病患者.收集所有参与本次研究患者的临床资料,包括年龄、患病类型、住院天数、中性粒细胞值、粒细胞缺乏时间、治疗方式、病原菌检出情况及药敏试验结果等,分析血液病患者发生血流感染的危险因素.血液病患者疑似发生血流感染后,在使用抗菌药物治疗前,抽取患者外周静脉血于血培养瓶内,采用全自动血培养仪进行细菌培养,采用全自动细菌鉴定仪进行病原菌鉴定及药敏试验.结果 150例发生血流感染的血液病患者,共检出病原菌148株.其中革兰阴性菌93株,革兰阳性菌44株,真菌11株.革兰阴性菌主要为肺炎克雷伯菌(21.62％),大肠埃希菌(16.22％),铜绿假单胞菌(10.81％).革兰阳性菌主要为凝固酶阴性葡萄球菌(12.84％)与金黄色葡萄球菌(7.43％).真菌均为热带念珠菌.药敏结果显示,肺炎克雷伯菌对头孢呋辛、头孢他啶、头孢曲松、氨曲南、复方新诺明的耐药率高于40％,对亚胺培南、美罗培南的耐药率低于20％.大肠埃希菌对头孢呋辛、头孢他啶、头孢曲松、环丙沙星、左氧氟沙星、复方新诺明的耐药率高于50％,对亚胺培南、美罗培南的耐药率低于10％.铜绿假单胞菌对头孢他啶、环丙沙星、左氧氟沙星、莫西沙星的耐药率高于40％.肺炎克雷伯菌、大肠埃希菌及铜绿假单胞菌均未产生对阿米卡星的耐药株.凝固酶阴性葡萄球菌对氨苄西林、红霉素、环丙沙星、左氧氟沙星、复方新诺明的耐药率高于50％,金黄色葡萄球菌对氨苄西林、红霉素、环丙沙星的耐药率高于40％,均未产生对万古霉素、利奈唑胺的耐药菌.对比150例发生血流感染与60例未发生血流感染的血液病患者临床资料,结果显示,年龄、中性粒细胞值、粒细胞缺乏时间、是否有侵入性操作、抗感染药物使用种类、是否接受化疗、是否使用糖皮质激素差异有统计学意义(P＜0.05).进一步进行二元Logistic分析发现,年龄＞60岁、中性粒细胞值≤0.2X 109/L、粒细胞缺乏时间＞7 d、抗感染药物使用种类＞2种、接受化疗、使用糖皮质激素是血液病合并血流感染的独立危险因素(P＜0.05).结论 血液病患者并发血流感染的病原菌主要为肺炎克雷伯菌、大肠埃希菌、凝固酶阴性葡萄球菌,对多种药物均呈现较高耐药率.发生血流感染的危险因素较多,临床上应采取有效措施积极预防.
Objective To predict the biochemical characteristics, structure and antigenic epitopes of Echinococcus multilocularis HSP90（EmHSP90）,and to provide a theoretical basis for the study of peptide vaccine against alveolar Echinococcosis. Methods Log in NCBI database to search and download the amino acid sequence of EmHSP90.The physical and chemical properties of EmHSP90 were predicted by Expasy onlineprediction website.SOPMA in PRABI database was used to predict the secondary structure of EmHSP90.SWISS-MODEL online prediction site was used to predict the tertiary structure of EmHSP90.The online tool NetAcet-1.0 predicted the acetylation site of EmHSP90,Netphos3.1 predicted the phosphorylation site of EmHSP90,and NetNGlyc1.0 predicted the n-glycosylation site of EmHSP90.Prediction of transmembrane domain of EmHSP90 by online database TMHMM.The Signal peptide sequence of EmHSP90 was predicted by Signal P-5.0 Serveronline database.Predicting Antigenic Peptides from an online database predicts the antigenic determinants of EmHSP90.Prediction of dominant B cell epitopes of EmHSP90 using IEDB database.Prediction of T cell dominance epitopes of EmHSP90 using SYFPEITHI online database. Results EmHSP90 was composed of 711 amino acids, its relative molecular weight was 80.049 24×10~3,and its instability coefficient was 41.09. Its average hydrophilic coefficient is-0.276,which is a hydrophilic protein.In the secondary structure, α-helix accounted for 50.91%,β-Angle accounted for 4.92%,extended chain accounted for 13.92%,random curling accounted for 30.24%. It contains 1 acetylation site, 64 phosphorylation sites（40 serine（Ser） phosphorylation sites, 16 threonine（Thr） phosphorylation sites, 8 tyrosine（Tyr） phosphorylation sites） and 4 n-glycosylation sites.It does not contain signal peptide sequences and transmembrane domains. The mean antigen propensity index of EmHSP90 was 1.024 4,and there were 27 epitope regions.There were 10 dominant epitopes in B cells, 13 dominant epitopes in CTL cells and 14 dominant epitopes in Th cells. Conclusion Bioinformatics predicted that EmHSP90 was an unstable hydrophilic protein containing B-cell and T-cell dominant epitopes, which could provide reference for the development of peptide vaccine against alveolar hydatid disease.
目的 分析武汉市2021 年分离乙型流感病毒基因组变异情况及分子进化特征.方法 采集2021 年武汉市流感样病例咽拭子标本,进行核酸检测、病毒分离,选取34 株乙型流感病毒代表毒株进行全基因组测序,利用生物信息学软件构建进化树并分析分子进化特征.结果 2021 年武汉市共监测病例标本2 242 例,流感病毒核酸检测阳性218例,均为乙型流感病毒Victoria 系(Influenza B virus Lineage Victoria,BV),阳性率9.72％.2021 年武汉分离的BV 系流感病毒与疫苗株比,HA 基因同源性为98.65％-99.24％,NA 基因同源性为98.89％-99.54％.2021 年武汉分离的BV系流感病毒归属于V1A.3a.1 和V1A.3a.2 分支,V1A.3a.2 为2021 年武汉市优势株.发现多处关键抗原表位发生了变异,未发现耐药位点变异.结论 2021 年武汉分离的BV 系流感病毒较疫苗株均存在数个抗原位点变异,因此应加强对病毒变异的持续监测,为疫苗株更新、流感防控及临床诊疗措施的制定提供参考.
