During 2018–2019,a severe human adenovirus(HAdV)infection outbreak occurred in southern China.Here,we screened 18 respiratory pathogens in 1704 children(≤14 years old)hospitalized with acute respiratory illness in G...During 2018–2019,a severe human adenovirus(HAdV)infection outbreak occurred in southern China.Here,we screened 18 respiratory pathogens in 1704 children(≤14 years old)hospitalized with acute respiratory illness in Guangzhou,China,in 2019.In total,151 patients had positive HAdV test results;34.4%(52/151)of them exhibited severe illness.HAdV infection occurred throughout the year,with a peak in summer.The median patient age was 3.0(interquartile range:1.1–5.0)years.Patients with severe HAdV infection exhibited increases in12 clinical indexes(P≤0.019)and decreases in four indexes(P≤0.007),compared with patients exhibiting nonsevere infection.No significant differences were found in age or sex distribution according to HAdV infection severity(P>0.05);however,the distributions of comorbid disease and HAdV co-infection differed according to HAdV infection severity(P<0.05).The main epidemic types were HAdV-3(47.0%,71/151)and HAdV-7(46.4%,70/151).However,the severe illness rate was significantly higher in patients with HAdV-7(51.4%)than in patients with HAdV-3(19.7%)and other types of HAdV(20%)(P<0.001).Sequencing analysis of genomes/capsid genes of 13 HAdV-7 isolates revealed high similarity to previous Chinese isolates.A representative HAdV-7isolate exhibited a similar proliferation curve to the curve described for the epidemic HAdV-3 strain Guangzhou01(accession no.DQ099432)(P>0.05);the HAdV-7 isolate exhibited stronger virulence and infectivity,compared with HAdV-3(P<0.001).Overall,comorbid disease,HAdV co-infection,and high virulence and infectivity of HAdV-7 were critical risk factors for severe HAdV infection;these data can facilitate treatment,control,and prevention of HAdV infection.展开更多
为了从分子水平上初步探讨我国流行HAdV-4的基因特征以及分子进化规律,为HAdV-4在我国的疫情监测与防控以及疫苗研发提供科学数据,本研究通过对2013～2014年间在陕西、云南、吉林和山东4省发热呼吸道症候群监测病例鼻咽拭子标本中分离...为了从分子水平上初步探讨我国流行HAdV-4的基因特征以及分子进化规律,为HAdV-4在我国的疫情监测与防控以及疫苗研发提供科学数据,本研究通过对2013～2014年间在陕西、云南、吉林和山东4省发热呼吸道症候群监测病例鼻咽拭子标本中分离得到的HAdV-4毒株,分别进行Penton base,Hexon和Fiber三个靶基因的序列测定,同时下载GenBank数据库中所有我国及全球流行HAdV-4病毒株的上述三基因序列进行比对,构建基因亲缘性关系树,并使用生物信息学分析软件进行基因特征和分子进化分析。结果显示:全球HAdV-4病毒株可划分两个进化分支(HAdV-4p和HAdV-4a),我国自2008年至今流行的HAdV-4病毒株属于HAdV-4a进化分支,与全球优势流行株一致;HAdV-4p和HAdV-4a进化分支病毒株的三个基因序列在时间和空间上均具保守性和稳定性,核苷酸和氨基酸突变主要与不同进化分支密切相关;HAdV-4基于三个基因的估算进化速率分别为3.73×10-5碱基替代/位点/年,1.86×10-5碱基替代/位点/年和4.09×10-5碱基替代/位点/年;基于Fiber基因估算的最近共同祖先(The most recent common ancestor,tMRCA)结果显示,HAdV-4起源于1771年,其中HAdV-4p和HAdV-4a可分别追溯至1933年和1962年。为了进一步积累并完善上述HAdV病毒学监测数据,我国急需建立长期有效的HAdV监测体系。展开更多
目的基于实时细胞分析(real time cell analysis,RTCA)技术对中药挥发性组分的细胞毒性及抗人腺病毒3型(human adenovirus 3,HAdV-3)的作用进行毒/效整合分析,构建抗病毒药物高通量筛选的新策略。方法采用RTCA技术动态监测不同接种密度...目的基于实时细胞分析(real time cell analysis,RTCA)技术对中药挥发性组分的细胞毒性及抗人腺病毒3型(human adenovirus 3,HAdV-3)的作用进行毒/效整合分析,构建抗病毒药物高通量筛选的新策略。方法采用RTCA技术动态监测不同接种密度的A549细胞48 h生长曲线及10倍梯度稀释的HAdV-3感染A549细胞生长曲线,获得A549细胞最佳接种密度及HAdV-3最佳稀释浓度用于后续实验。以利巴韦林为阳性对照,基于RTCA技术采用曲线下面积(Area Under the Curve,AUC)方法对5种中药挥发性组分的细胞毒性及抗HAdV-3的作用进行毒/效整合分析,并与传统终点法数据处理进行对比。结果终点法中艾叶、柴胡、薄荷、荆芥、牛蒡子、酸枣仁和利巴韦林对细胞的保护率分别为0.73%、12.50%、20.99%、44.50%、27.99%、51.50%和82.70%;AUC法中艾叶、柴胡、薄荷的选择性指数(Selective Index,SI)为负数,分别为-0.57、-0.21和-0.08。荆芥、牛蒡子、酸枣仁和利巴韦林的SI值分别为0.14、0.40、0.72和5.33。以利巴韦林的抗病毒活性为参照,终点法中酸枣仁、牛蒡子和荆芥的抗病毒活性为利巴韦林抗病毒活性的62.27%、33.85%和53.81%;AUC法中酸枣仁、牛蒡子和荆芥的抗病毒活性为利巴韦林抗病毒活性的13.51%、7.50%和2.63%。结论传统终点法与研究采用的AUC法计算结果有较大的差异。传统的抗病毒活性筛选研究多采用终点法检测受试药物对病毒感染细胞的保护作用,无法反映整个感染周期的变化,缺乏细胞毒的数据也会对结果的判断产生偏差。研究提示基于RTCA技术对中药挥发性组分开展抗HAdV-3的毒/效整合评价,采用AUC方法计算药物的选择性指数作为高通量筛选新策略,能快速、精准地判断受试药物的抗病毒活性,具有准确性高、重复性好的优势。展开更多
基金supported by the Guangzhou Science and Technology Program-Zhongnanshan Medical Foundation of Guangdong Province(202102010359-ZNSA-2020003)the Emergency Key Program of Guangzhou Laboratory(EKPG21-13)+3 种基金the National Natural Science Foundation of China(81970003,31900877)the Natural Science Foundation of Guangdong Province of China(2018A030310401)Guangdong-Hong Kong-Macao Joint Laboratory of Respiratory Infectious Disease(GHMJLRID-Z-202109)the Special Project for COVID-19 Prevention and Control of Zhongnanshan Medical Foundation of Guangdong Province(ZNSA-2020012)。
文摘During 2018–2019,a severe human adenovirus(HAdV)infection outbreak occurred in southern China.Here,we screened 18 respiratory pathogens in 1704 children(≤14 years old)hospitalized with acute respiratory illness in Guangzhou,China,in 2019.In total,151 patients had positive HAdV test results;34.4%(52/151)of them exhibited severe illness.HAdV infection occurred throughout the year,with a peak in summer.The median patient age was 3.0(interquartile range:1.1–5.0)years.Patients with severe HAdV infection exhibited increases in12 clinical indexes(P≤0.019)and decreases in four indexes(P≤0.007),compared with patients exhibiting nonsevere infection.No significant differences were found in age or sex distribution according to HAdV infection severity(P>0.05);however,the distributions of comorbid disease and HAdV co-infection differed according to HAdV infection severity(P<0.05).The main epidemic types were HAdV-3(47.0%,71/151)and HAdV-7(46.4%,70/151).However,the severe illness rate was significantly higher in patients with HAdV-7(51.4%)than in patients with HAdV-3(19.7%)and other types of HAdV(20%)(P<0.001).Sequencing analysis of genomes/capsid genes of 13 HAdV-7 isolates revealed high similarity to previous Chinese isolates.A representative HAdV-7isolate exhibited a similar proliferation curve to the curve described for the epidemic HAdV-3 strain Guangzhou01(accession no.DQ099432)(P>0.05);the HAdV-7 isolate exhibited stronger virulence and infectivity,compared with HAdV-3(P<0.001).Overall,comorbid disease,HAdV co-infection,and high virulence and infectivity of HAdV-7 were critical risk factors for severe HAdV infection;these data can facilitate treatment,control,and prevention of HAdV infection.
文摘为了从分子水平上初步探讨我国流行HAdV-4的基因特征以及分子进化规律,为HAdV-4在我国的疫情监测与防控以及疫苗研发提供科学数据,本研究通过对2013～2014年间在陕西、云南、吉林和山东4省发热呼吸道症候群监测病例鼻咽拭子标本中分离得到的HAdV-4毒株,分别进行Penton base,Hexon和Fiber三个靶基因的序列测定,同时下载GenBank数据库中所有我国及全球流行HAdV-4病毒株的上述三基因序列进行比对,构建基因亲缘性关系树,并使用生物信息学分析软件进行基因特征和分子进化分析。结果显示:全球HAdV-4病毒株可划分两个进化分支(HAdV-4p和HAdV-4a),我国自2008年至今流行的HAdV-4病毒株属于HAdV-4a进化分支,与全球优势流行株一致;HAdV-4p和HAdV-4a进化分支病毒株的三个基因序列在时间和空间上均具保守性和稳定性,核苷酸和氨基酸突变主要与不同进化分支密切相关;HAdV-4基于三个基因的估算进化速率分别为3.73×10-5碱基替代/位点/年,1.86×10-5碱基替代/位点/年和4.09×10-5碱基替代/位点/年;基于Fiber基因估算的最近共同祖先(The most recent common ancestor,tMRCA)结果显示,HAdV-4起源于1771年,其中HAdV-4p和HAdV-4a可分别追溯至1933年和1962年。为了进一步积累并完善上述HAdV病毒学监测数据,我国急需建立长期有效的HAdV监测体系。
文摘目的基于实时细胞分析(real time cell analysis,RTCA)技术对中药挥发性组分的细胞毒性及抗人腺病毒3型(human adenovirus 3,HAdV-3)的作用进行毒/效整合分析,构建抗病毒药物高通量筛选的新策略。方法采用RTCA技术动态监测不同接种密度的A549细胞48 h生长曲线及10倍梯度稀释的HAdV-3感染A549细胞生长曲线,获得A549细胞最佳接种密度及HAdV-3最佳稀释浓度用于后续实验。以利巴韦林为阳性对照,基于RTCA技术采用曲线下面积(Area Under the Curve,AUC)方法对5种中药挥发性组分的细胞毒性及抗HAdV-3的作用进行毒/效整合分析,并与传统终点法数据处理进行对比。结果终点法中艾叶、柴胡、薄荷、荆芥、牛蒡子、酸枣仁和利巴韦林对细胞的保护率分别为0.73%、12.50%、20.99%、44.50%、27.99%、51.50%和82.70%;AUC法中艾叶、柴胡、薄荷的选择性指数(Selective Index,SI)为负数,分别为-0.57、-0.21和-0.08。荆芥、牛蒡子、酸枣仁和利巴韦林的SI值分别为0.14、0.40、0.72和5.33。以利巴韦林的抗病毒活性为参照,终点法中酸枣仁、牛蒡子和荆芥的抗病毒活性为利巴韦林抗病毒活性的62.27%、33.85%和53.81%;AUC法中酸枣仁、牛蒡子和荆芥的抗病毒活性为利巴韦林抗病毒活性的13.51%、7.50%和2.63%。结论传统终点法与研究采用的AUC法计算结果有较大的差异。传统的抗病毒活性筛选研究多采用终点法检测受试药物对病毒感染细胞的保护作用,无法反映整个感染周期的变化,缺乏细胞毒的数据也会对结果的判断产生偏差。研究提示基于RTCA技术对中药挥发性组分开展抗HAdV-3的毒/效整合评价,采用AUC方法计算药物的选择性指数作为高通量筛选新策略,能快速、精准地判断受试药物的抗病毒活性,具有准确性高、重复性好的优势。
基金A Project from the Hospital (Grant No. 2021-XZYG-B11)the Key Research and Development Project of the Sichuan Science and Technology Department (Grant No. 2019YFS0367)
