目的对蜱传疾病患者流行病学资料、临床特征进行分析,以期为蜱传疾病诊治能力提升提供帮助。方法收集北京地坛医院2013年1月至2023年6月收治的蜱传疾病患者资料,对流行病学和临床特征进行分析。结果共纳入32例蜱传疾病患者临床资料,包...目的对蜱传疾病患者流行病学资料、临床特征进行分析,以期为蜱传疾病诊治能力提升提供帮助。方法收集北京地坛医院2013年1月至2023年6月收治的蜱传疾病患者资料,对流行病学和临床特征进行分析。结果共纳入32例蜱传疾病患者临床资料,包括发热伴血小板减少综合征(severe fever with thrombocytopenia syndrome,SFTS)17例,莱姆病5例,立克次体病10例。发病时间集中在每年的5—10月。北京本土病例14例(43.8%),平谷区占比最高(4例,28.6%),外省输入性病例16例(50.0%),境外旅居患病人员2例(6.3%)。病例年龄21~80岁,平均(50.16±14.61)岁。病例数最多的职业为农民(14例,43.8%)。游走性红斑为莱姆病典型皮损特点,发热多见于SFTS及立克次体病患者,以中度发热为主。WBC及NEUT降低多见于SFTS。SFTS及立克次体病均有EOS降低。PLT减少为SFTS典型特征。立克次体病及SFTS患者表现出明显的肝脏损伤,ALT、AST明显升高。SFTS患者存在明显心肌损伤,CK、CK-MB均有升高。CRP升高主要见于立克次体病。PCT升高最常见于立克次体病,其次为SFTS。结论北京地区蜱传疾病存在逐年上升趋势,多有延误诊断,需提升此类疾病的诊断能力,根据流行病学史和蜱传疾病临床特征进行早期识别,及早行病原学检测,从而提高早期诊断率,实现早诊早治。展开更多
Objective The purpose of this study was to investigate the bacterial communities of biting midges and ticks collected from three sites in the Poyang Lake area,namely,Qunlu Practice Base,Peach Blossom Garden,and Huangt...Objective The purpose of this study was to investigate the bacterial communities of biting midges and ticks collected from three sites in the Poyang Lake area,namely,Qunlu Practice Base,Peach Blossom Garden,and Huangtong Animal Husbandry,and whether vectors carry any bacterial pathogens that may cause diseases to humans,to provide scientific basis for prospective pathogen discovery and disease prevention and control.Methods Using a metataxonomics approach in concert with full-length 16S rRNA gene sequencing and operational phylogenetic unit(OPU)analysis,we characterized the species-level microbial community structure of two important vector species,biting midges and ticks,including 33 arthropod samples comprising 3,885 individuals,collected around Poyang Lake.Results A total of 662 OPUs were classified in biting midges,including 195 known species and 373 potentially new species,and 618 OPUs were classified in ticks,including 217 known species and 326 potentially new species.Surprisingly,OPUs with potentially pathogenicity were detected in both arthropod vectors,with 66 known species of biting midges reported to carry potential pathogens,including Asaia lannensis and Rickettsia bellii,compared to 50 in ticks,such as Acinetobacter lwoffii and Staphylococcus sciuri.We found that Proteobacteria was the most dominant group in both midges and ticks.Furthermore,the outcomes demonstrated that the microbiota of midges and ticks tend to be governed by a few highly abundant bacteria.Pantoea sp7 was predominant in biting midges,while Coxiella sp1 was enriched in ticks.Meanwhile,Coxiella spp.,which may be essential for the survival of Haemaphysalis longicornis Neumann,were detected in all tick samples.The identification of dominant species and pathogens of biting midges and ticks in this study serves to broaden our knowledge associated to microbes of arthropod vectors.Conclusion Biting midges and ticks carry large numbers of known and potentially novel bacteria,and carry a wide range of potentially pathogenic bacteria,which may pose a risk of infection to humans and animals.The microbial communities of midges and ticks tend to be dominated by a few highly abundant bacteria.展开更多
目的:制备粉尘螨变应原第27组分(Der f 27)重组蛋白,鉴定其免疫活性。方法:构建pET-28a(+)-Der f 27质粒,转化至E.coli BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 27。IgE-ELISA和IgE-Western blot检测rDer f 27与...目的:制备粉尘螨变应原第27组分(Der f 27)重组蛋白,鉴定其免疫活性。方法:构建pET-28a(+)-Der f 27质粒,转化至E.coli BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 27。IgE-ELISA和IgE-Western blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率,分别将变应性鼻炎患者PBMC、人支气管上皮细胞BESA-2B与rDer f 27共孵育24 h后测定细胞因子表达;生物信息学软件分析Der f 27的理化性质和结构。结果:成功构建pET-28a(+)-Der f 27质粒并转化至BL21(DE3)细胞,经IPTG诱导表达和纯化后,SDS-PAGE和Western blot在约48 kD处见特异性条带;IgE-ELISA和IgEWestern blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率分别为39.5%和45.5%;与对照组相比,变应性鼻炎患者PBMC与rDer f 27共孵育后,IL-6和IL-8表达升高(P<0.05);BESA-2B细胞与rDer f 27共孵育后,IL-10和TGF-β表达降低,IL-17A和IL-23A表达升高(P<0.05);生物信息学结果显示Der f 27属于丝氨酸蛋白酶抑制物家族,具有此家族通用结构及功能,二级结构主要为α螺旋(42.62%)和无规则卷曲(35.60%)。结论:成功制备了具有较好免疫反应性和免疫原性的重组变应原rDer f 27,并揭示其为变应性鼻炎的重要变应原之一。展开更多
目的:制备粉尘螨变应原第36组分(Der f 36)重组蛋白,鉴定其免疫原性并进行生物信息学分析。方法:获得Der f 36核酸序列并进行人工合成,插入pET-28a(+)载体,制备pET-28a(+)-Der f 36质粒,转化至BL21(DE3)感受态细胞,经诱导表达和纯化后,...目的:制备粉尘螨变应原第36组分(Der f 36)重组蛋白,鉴定其免疫原性并进行生物信息学分析。方法:获得Der f 36核酸序列并进行人工合成,插入pET-28a(+)载体,制备pET-28a(+)-Der f 36质粒,转化至BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 36,SDS-PAGE和Western blot鉴定rDer f 36重组蛋白,IgE-ELISA检测其血清IgE结合率,IgE-ELISA抑制实验检测rDer f 36与rDer p 36的交叉反应性;HNEpC细胞与rDer f 36共孵育24 h后测定细胞因子;生物信息学软件比较Der f 36与Der p 36的理化性质和结构。结果:得到了Der f 36的编码基因,全长690 bp,相对分子质量为25.6 kD;IgE-ELISA鉴定rDer f 36血清IgE结合率为42.1%;IgE-ELISA抑制实验结果显示rDer p 36对rDer f 36的抑制率>50.00%[40.00%(8/20)],平均抑制率为52.98%。HNEpC细胞与rDer f 36共孵育后,IL-6、IL-8、IL-33、IL-25和TSLP表达均高于对照组。生物信息学分析显示Der f 36和Der p 36的序列一致性为77.63%,理化性质较为相似;二级结构分析结果显示二者均含有α螺旋、β转角和无规则卷曲,且无规则卷曲含量最高;C2结构域显示高度重叠(RMSD=0.046)。结论:成功制备了rDer f 36重组蛋白,且具有免疫原性,为粉尘螨变应原单组分诊断及治疗奠定了基础。Der f 36与Der p 36的理化性质、二级结构和三级结构高度相似,决定其具有交叉反应性。展开更多
文摘目的对蜱传疾病患者流行病学资料、临床特征进行分析,以期为蜱传疾病诊治能力提升提供帮助。方法收集北京地坛医院2013年1月至2023年6月收治的蜱传疾病患者资料,对流行病学和临床特征进行分析。结果共纳入32例蜱传疾病患者临床资料,包括发热伴血小板减少综合征(severe fever with thrombocytopenia syndrome,SFTS)17例,莱姆病5例,立克次体病10例。发病时间集中在每年的5—10月。北京本土病例14例(43.8%),平谷区占比最高(4例,28.6%),外省输入性病例16例(50.0%),境外旅居患病人员2例(6.3%)。病例年龄21~80岁,平均(50.16±14.61)岁。病例数最多的职业为农民(14例,43.8%)。游走性红斑为莱姆病典型皮损特点,发热多见于SFTS及立克次体病患者,以中度发热为主。WBC及NEUT降低多见于SFTS。SFTS及立克次体病均有EOS降低。PLT减少为SFTS典型特征。立克次体病及SFTS患者表现出明显的肝脏损伤,ALT、AST明显升高。SFTS患者存在明显心肌损伤,CK、CK-MB均有升高。CRP升高主要见于立克次体病。PCT升高最常见于立克次体病,其次为SFTS。结论北京地区蜱传疾病存在逐年上升趋势,多有延误诊断,需提升此类疾病的诊断能力,根据流行病学史和蜱传疾病临床特征进行早期识别,及早行病原学检测,从而提高早期诊断率,实现早诊早治。
基金supported by grants from National Key R&D Program of China(2019YFC1200501)Research Units of Discovery of Unknown Bacteria and Function(2018RU010)Capacity Enhancement Project supported by National Institute for Communicable Disease Control and Prevention(China CDC).
文摘Objective The purpose of this study was to investigate the bacterial communities of biting midges and ticks collected from three sites in the Poyang Lake area,namely,Qunlu Practice Base,Peach Blossom Garden,and Huangtong Animal Husbandry,and whether vectors carry any bacterial pathogens that may cause diseases to humans,to provide scientific basis for prospective pathogen discovery and disease prevention and control.Methods Using a metataxonomics approach in concert with full-length 16S rRNA gene sequencing and operational phylogenetic unit(OPU)analysis,we characterized the species-level microbial community structure of two important vector species,biting midges and ticks,including 33 arthropod samples comprising 3,885 individuals,collected around Poyang Lake.Results A total of 662 OPUs were classified in biting midges,including 195 known species and 373 potentially new species,and 618 OPUs were classified in ticks,including 217 known species and 326 potentially new species.Surprisingly,OPUs with potentially pathogenicity were detected in both arthropod vectors,with 66 known species of biting midges reported to carry potential pathogens,including Asaia lannensis and Rickettsia bellii,compared to 50 in ticks,such as Acinetobacter lwoffii and Staphylococcus sciuri.We found that Proteobacteria was the most dominant group in both midges and ticks.Furthermore,the outcomes demonstrated that the microbiota of midges and ticks tend to be governed by a few highly abundant bacteria.Pantoea sp7 was predominant in biting midges,while Coxiella sp1 was enriched in ticks.Meanwhile,Coxiella spp.,which may be essential for the survival of Haemaphysalis longicornis Neumann,were detected in all tick samples.The identification of dominant species and pathogens of biting midges and ticks in this study serves to broaden our knowledge associated to microbes of arthropod vectors.Conclusion Biting midges and ticks carry large numbers of known and potentially novel bacteria,and carry a wide range of potentially pathogenic bacteria,which may pose a risk of infection to humans and animals.The microbial communities of midges and ticks tend to be dominated by a few highly abundant bacteria.
文摘目的:制备粉尘螨变应原第27组分(Der f 27)重组蛋白,鉴定其免疫活性。方法:构建pET-28a(+)-Der f 27质粒,转化至E.coli BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 27。IgE-ELISA和IgE-Western blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率,分别将变应性鼻炎患者PBMC、人支气管上皮细胞BESA-2B与rDer f 27共孵育24 h后测定细胞因子表达;生物信息学软件分析Der f 27的理化性质和结构。结果:成功构建pET-28a(+)-Der f 27质粒并转化至BL21(DE3)细胞,经IPTG诱导表达和纯化后,SDS-PAGE和Western blot在约48 kD处见特异性条带;IgE-ELISA和IgEWestern blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率分别为39.5%和45.5%;与对照组相比,变应性鼻炎患者PBMC与rDer f 27共孵育后,IL-6和IL-8表达升高(P<0.05);BESA-2B细胞与rDer f 27共孵育后,IL-10和TGF-β表达降低,IL-17A和IL-23A表达升高(P<0.05);生物信息学结果显示Der f 27属于丝氨酸蛋白酶抑制物家族,具有此家族通用结构及功能,二级结构主要为α螺旋(42.62%)和无规则卷曲(35.60%)。结论:成功制备了具有较好免疫反应性和免疫原性的重组变应原rDer f 27,并揭示其为变应性鼻炎的重要变应原之一。
文摘目的:制备粉尘螨变应原第36组分(Der f 36)重组蛋白,鉴定其免疫原性并进行生物信息学分析。方法:获得Der f 36核酸序列并进行人工合成,插入pET-28a(+)载体,制备pET-28a(+)-Der f 36质粒,转化至BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 36,SDS-PAGE和Western blot鉴定rDer f 36重组蛋白,IgE-ELISA检测其血清IgE结合率,IgE-ELISA抑制实验检测rDer f 36与rDer p 36的交叉反应性;HNEpC细胞与rDer f 36共孵育24 h后测定细胞因子;生物信息学软件比较Der f 36与Der p 36的理化性质和结构。结果:得到了Der f 36的编码基因,全长690 bp,相对分子质量为25.6 kD;IgE-ELISA鉴定rDer f 36血清IgE结合率为42.1%;IgE-ELISA抑制实验结果显示rDer p 36对rDer f 36的抑制率>50.00%[40.00%(8/20)],平均抑制率为52.98%。HNEpC细胞与rDer f 36共孵育后,IL-6、IL-8、IL-33、IL-25和TSLP表达均高于对照组。生物信息学分析显示Der f 36和Der p 36的序列一致性为77.63%,理化性质较为相似;二级结构分析结果显示二者均含有α螺旋、β转角和无规则卷曲,且无规则卷曲含量最高;C2结构域显示高度重叠(RMSD=0.046)。结论:成功制备了rDer f 36重组蛋白,且具有免疫原性,为粉尘螨变应原单组分诊断及治疗奠定了基础。Der f 36与Der p 36的理化性质、二级结构和三级结构高度相似,决定其具有交叉反应性。
