Objective:To analyze the dynamic changes and possible influencing factors of anti-2019 novel Coronavirus (2019-nCoV) neutralizing antibody in confirmed Coronavirus Disease 2019 (COVID-19) cases.Methods:Microneutralization was used to test the anti-2019-nCoV neutralizing antibody. Excel 2007 and SPSS 22.0 were used for data processing and analysis.Results:There were 420 serum samples collected from 155 confirmed COVID-19 cases. These serum samples contained acute phase serum, convalescent phase serum and serum from cases recovered for about six months. The sampling time was 0-221 days after the onset of COVID-19. The geometric mean titer (GMT) of anti-2019-nCoV neutralizing antibody was 1∶13 at 1 week, and 1∶31 at 2 week. The titers of anti-2019-nCoV neutralizing antibody of individual cases were still<1∶4 on the 15 th day. The GMT was all over 1: 52 (13×4) at 6-32 week. Taking 1: 64 as the cut-off point, the serum anti-2019-nCoV neutralizing antibody positive rates was 30.56% in acute phase serum samples (0-14 d, 0-2 w), 82.31% in convalescent phase serum samples (36-63 d, 6-9 w) and 86.52% in serum samples from cases recovered for about six months (183-210 d, 27-30 w). Statistical analysis showed that there was no significant difference in anti-2019-nCoV neutralizing antibody levels at the other weeks except 1-2 week ( χ2=9.270, P=0.931), there was no statistically differences in gender, age and occupation of the cases, and also between the normal and mild cases ( P>0.05). Conclusions:The serum anti-2019-nCoV neutralizing antibody level is only statistically correlated with the disease progression of COVID-19, and maintain the protective level from 3 to 30 week.

Severe fever with thrombocytopenia syndrome (SFTS) is recognized as an emerging infectious disease. This study aimed to investigate the pathogenic mechanism of SFTS. A total of 100 subjects were randomly included in the study. Cytokine levels were detected by enzyme‐linked immunosorbent assay and the viral load was detected by micro drop digital PCR. The results showed that levels of interleukin‐6 (IL‐6), IL‐8, IL‐10, IFN‐inducible protein‐10 (IP‐10), monocyte chemoattractant protein‐1 (MCP‐1), macrophage inflammatory protein‐1α (MIP‐1α), transforming growth factor‐β1 (TGF‐β1), and regulated upon activation normal T cell expressed and secreted factor (RANTES) differed significantly among the SFTS patient group, healthy people group, and asymptomatic infection group (p < .05). Compared to the healthy people group, the patient group had increased cytokine levels (IL‐6, IL‐10, IP‐10, MCP‐1, and IFN‐γ) but reduced levels of IL‐8, TGF‐β1, and RANTES (p < .0167). IL‐6, IL‐8, IL‐10, IP‐10, MCP‐1, MIP‐1α, TGF‐β1, and the RANTES levels had different trends after the onset of the disease. IL‐6, IL‐10, IP‐10, and MCP‐1 levels in severe patients were higher than those in mild patients (p < .05). There was a positive correlation between viral load and IL‐6 and IP‐10 but a negative correlation between viral load and RANTES. SFTSV could cause a cytokine change: the cytokine levels of patients had different degrees of fluctuation after the onset of the disease. The levels of IL‐6 and IL‐8 in the asymptomatic infection group were found between the SFTS patients group and the healthy people group. The levels of IL‐6, IL‐10, IP‐10, and MCP‐1 in the serum could reflect the severity of the disease, and the levels of IL‐6, IP‐10, and RANTES were correlated with the viral load.

目的:回顾新型冠状病毒(2019-nCoV)合并Ⅰ型单纯疱疹病毒(HSV-1)感染的诊断方法.方法:收集12例新型冠状病毒肺炎确诊病例的核酸阳性咽拭子样本,采用Vero和Vero E6细胞进行病毒分离,采用实时荧光定量PCR法进行2019-nCoV核酸检测,采用MiSeq测序技术测定分离株基因组序列,采用MicroSpectrum病原体鉴定分析软件分析非2019-nCoV毒株组分.结果:12例12份咽拭子样本中分离得到5株病毒,其中4株病毒各代培养物均呈2019-nCoV核酸阳性,HN03株在培养过程中观察到支原体感染表现并逐渐表现为2019-nCoV核酸阴性.MiSeq测序获得4株2019-nCoV全基因组序列,HN03株获得部分2019-nCoV基因序列.HN03株的三代培养物经宏基因组序列测定、分析,结果提示HN03株为HSV-1和猪鼻支原体双阳性.结论:HN03株源病例为2019-nCoV和HSV-1合并感染.

目的 分离新型冠状病毒毒株,分析新型冠状病毒基本生物学特征和遗传进化特征.方法 筛选具有代表性的新型冠状病毒核酸阳性样本,采用Vero和Vero-E6细胞系分离病毒、荧光定量RT-PCR方法初步鉴定,MiSeq测序技术获取新型冠状病毒全基因组序列,开展进化分析.结果 4个新型冠状病毒核酸阳性的咽拭子样本在Vero和Vero E6细胞均产生细胞病变,细胞病变可以稳定传代.荧光定量RT-PCR鉴定四株病毒二代培养物均显示为新型冠状病毒核酸阳性;三株病毒二代培养物经Miseq测序获得新型冠状病毒全基因组序列,一株病毒二代培养物获得部分新型冠状病毒基因组序列.全基因序列比较显示三株新型冠状病毒均与武汉分离株Wuhan-Hu-1和WIV04亲缘关系最为接近,HN01与Wuhan-Hu-1株的NS3蛋白存在一个氨基酸差异,HN02与Wuhan-Hu-1株的NS8蛋白存在一个氨基酸差异,HN04与Wuhan-Hu-1株在同源编码区核苷酸同源性为100％.结论 河南省新型冠状病毒株与湖北武汉株高度同源,暂未发现有意义的新型冠状病毒变异株.

Abstract Background Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease with the high case-fatality rate, and lack of vaccines. We aimed to systematically analysed the epidemiological characteristics, clinical signs, routine laboratory diagnosis, risk factors, and outcomes. Methods Documents on SFTS were collected by searching the Chinese National Knowledge Infrastructure, Wan Fang Data, PubMed, Embase, and Web of Science databases from 2011 to 2018. Meta-analysis was performed by using Review Manager and Stata software. Results Twenty-five articles involving 4143 cases were included. Diarrhea (odds ratio (OR) =1.60, 95% confidence interval (CI): 1.06 to 2.42, P = 0.02), and vomiting (OR = 1.56, 95% CI: 1.01 to 2.39, P = 0.04) on admission were associated with the fatal outcomes of SFTS. Compared to patients with mild symptoms, patients with severe symptoms had significantly elevated levels of lactic acid dehydrogenase (standard mean difference (SMD) =1.27, 95% CI: 0.59 to 1.94), alanine aminotransferase (SMD = 0.55, 95% CI: 0.24 to 0.85), aspirate aminotransferase (SMD = 1.01, 95% CI: 0.69 to 1.32), and creatine kinase (SMD = 1.04, 95% CI: 0.74 to 1.33) but had reduced platelet counts (SMD = -0.87, 95% CI: − 1.16 to − 0.58) and albumin levels (SMD = -1.00, 95% CI: − 1.32 to − 0.68). The risk factors for poor prognosis included age (mean difference (MD) =6.88, 95% CI: 5.41 to 8.35) and farming (OR = 2.01, 95% CI: 1.06 to 3.80). For the risk factors of contracting SFTS, the incidence of SFTS related to tick bites was 24% [95% CI: 0.18 to 0.31]. The pooled case-fatality rate of SFTS patients was 18% [95% CI: 0.16 to 0.21]. Conclusions China is the country with the highest incidence of SFTS. May to July was the peak of the epidemic, and farmers were a high-risk group. The risk factor for SFTS included age (poor prognosis) and tick bites (contracting SFTS). Patients with severe diarrhea and vomiting symptoms on admission should be noted. Clinicians could use routine laboratory parameters and clinical symptoms as references for clinically suspected cases, classification of SFTS, and timely treatment, especially in basic hospitals.

ObjectiveTo understand the epidemiological and etiological characteristics of an imported Chikungunya fever case occurred in Huixian county of Henan province in August 2019 and provide evidence for the prevention and control of the further spread. MethodsThe serum samples of the case, person with co-exposure and contacts as well as mosquito samples were detected with real-time fluorescence quantitative RT-PCR and the virus isolation was conducted, E1 gene of the isolate was sequenced and analyzed by Clustal X and MEGA 7.0. Epidemiological investigation was conducted, and prevention and control measures were taken according to related technical guidelines. ResultsThe serum sample of the case was positive for nucleic acid of Chikungunya virus. The virus isolate (GenBank No. MN432879) belonged to ECSA genotype of Chikungunya virus. In E1 section, the isolate shared 100% nucleotide/amino acid homology of MN432879 with an imported strain (GenBank No. MG912993) in Zhejiang in 2017, and shared 99.4% nucleotide homology and 99.8% amino acid homology with an imported strain (GenBank No. MG925665) isolated in Henan. There was one case with co-exposure and three contacts, all of them showed no clinical manifestations of Chikungunya fever. The breteau index of the epidemic point declined from 9.5 in the first surveillance to 3.2 in the third surveillance and remained at a low level. Mosquito samples were negative for nucleic acid of Chikungunya virus. ConclusionThe imported Chikungunya fever case in Huixian county of Henan was caused by ECSA genotype Chikungunya virus, no further spread occurred in this area.

目的:总结和分析河南省炭疽疫情概况及流行特征,为炭疽监测与防控工作提供科学依据.方法:采用描述性流行病学方法对河南省1956～2018年炭疽疫情数据资料进行分析.结果:1956～2018年河南省共报告人间炭疽6558例,均为皮肤炭疽,年均发病率0.21/10万;死亡258例,病死率3.93％.流行高峰出现在1957年,1960年代初和年代末又各有一次流行高峰,之后疫情总体呈现下降趋势.炭疽病例主要分布在南阳、新乡、洛阳、许昌、开封5个地区,占全省病例的89.60％.乡村发病率高于城市,盆地和山区炭疽发病率较高.炭疽病例以青壮年为多,男性多于女性.发病集中在6～9月,其中8月是流行高峰.牛、羊、猪、马等家畜是人间炭疽的传染源.病例潜伏期2～9 d,平均5 d.结论:河南省1950～1980年代是炭疽流行期,盆地及山区的农村地区是主要流行地区.

Objective To survey avian influenza A viruses (AIVs) in the environment and explore the reasons for the surge in human H7N9 cases. Methods A total of 1,045 samples were collected from routine surveillance on poultry-related environments and 307 samples from human H7N9 cases-exposed environments in Henan from 2016 to 2017. The nucleic acids of influenza A (Flu A), H5, H7, and H9 subtypes were detected by real-time polymerase chain reaction. Results A total of 27 H7N9 cases were confirmed in Henan from 2016 to 2017, 24 had a history of live poultry exposure, and 15 had H7N9 virus detected in the related live poultry markets (LPMs). About 96% (264/275) Flu A positive-environmental samples were from LPMs. H9 was the main AIV subtype (10.05%) from routine surveillance sites with only 1 H7-positive sample, whereas 21.17% samples were H7-positive in H7N9 cases-exposed environments. Samples from H7N9 cases-exposed LPMs (47.56%) had much higher AIVs positive rates than those from routine surveillance sites (12.34%). The H7+H9 combination of mixed infection was 78.18% (43/55) of H7-positive samples and 41.34% (43/104) of H9-positive samples. Conclusion The contamination status of AIVs in poultry-related environments is closely associated with the incidence of human infection caused by AIVs. Therefore, systematic surveillance of AIVs in LPMs in China is essential for the detection of novel reassortant viruses and their potential for interspecies transmission.

After we published our preliminary study on the use of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and curated E. coli toxin databases on the identification of E. coli Shiga toxins (Stxs) in the Journal of Proteomics in year 2018, we were encouraged to further refine the method and test clinical isolates. In this study, different concentrations of mitomycin C (MMC) and ciprofloxacin (CF), two common antibiotic/chemotherapy agents capable of stimulating Stx production, were first tested and compared on three reference strains and eight clinical isolates to observe the toxin induction and subsequent identification. Notably, no differences were observed between the two agents other than the concentrations applied. Seventeen more clinical isolates were then tested using fixed MMC and CF concentrations and sample amount. This study confirms that the majority of stx2-positive E. coli strains can be stimulated to produce sufficient toxin for confident identification. This does not occur with stx1-positive E. coli isolates, however, despite the fact that both Stxs can be identified for several isolates without MMC or CF stimulation. BIOLOGICAL SIGNIFICANCE: Stxs, especially Stx2, are very important causes of severe food-borne disease, even death. This study confirms that receptor analogue-based affinity enrichment of Stxs, after MMC or CF treatment of E. coli, is useful for fast and accurate Stx2 identification through LC-MS/MS.

To study the epidemiology and etiology characteristics of first imported Chikungunya fever case in Henan province, China, 2017. The patient was confirmed by Chikungunya virus (CHIKV) infected as CHIKV ribonucleotide was continuously detected in his serum specimens. BHK-21 cell line was used for virus isolation, the strain was named CHIKV/Henan001/2017. CHIKV/Henan001/2017 belonged to genotype ECSA. The highest ribonucleotide homology sequence of highly conserved region E1 with CHIKV/Henan001/2017 was hk02 strain (99.8%), who was an imported strain to Hong Kong, China, 2016. Epidemiological information and laboratory testing confirmed it was an imported Chikungunya fever case in Henan province, 2017. No secondary case has been reported.

Objective To investigate and analyze the clinical features,epidemiologic information and pathogenic characteristics of a rabies patient.Methods Clinical data of the patient(boy) was collected and epidemiological survey was conducted,fluorescence quantitative reverse transcription-polymerase chain reaction (FQRT-PCR) and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the samples of saliva,cerebrospinal fluid (CSF),skin tissue with hair follicle at the back of the neck for rabies laboratory diagnosis.Results Early symptoms of the boy were vomiting,diarrhea,fever and irritability,followed by coma and death.The boy had nasal trauma one month ago and the domestic dog died of illness during the same period.He did not accept the rabies post-exposure prophylaxis (PEP).The result of the saliva sample was positive by FQRT-PCR.The predicted segments of the glycoprotein (G),nucleoprotein (N) genes of rabies virus were amplified from the positive saliva sample of the patient by RT-PCR.Compared with rabies virus strains in Henan province,the nucleotide homology and amino acid homology in G gene segment were 96.5％-98.8％ and 96.5％-99.2％ respectively.Conclusions The case was diagnosed in laboratory as rabies case.The pathogenic rabies virus strain was endemic in Henan province.The nasal trauma,the dead domestic dog were probably related to the infection of the boy.

目的 应用人胚肺成纤维细胞MRC-5培养并分离鉴定寨卡病毒.方法 收集寨卡病毒病确诊病例的精液样本并在MRC-5细胞中进行培养,观察致细胞病变效应(CPE).采用实时荧光PCR (real-time PCR)方法鉴定培养,同时对培养物进行序列测定和比对分析.结果 病例精液样本经MRC-5细胞盲传3代,未观察到明显的特异性致CPE,MRC-5细胞3代培养物寨卡病毒经real-time PCR检测结果呈阳性.毒株命名为Henan/001/2016,GenBank编号为MF593625,属于亚洲基因簇系.Henan/001/2016与Natal RGN株的亲缘关系最为接近,在E基因区段的核苷酸和氨基酸序列同源性分别为99.7％和100％.结论 MRC-5细胞适用于寨卡病毒的分离、培养和鉴定.

Toxin expression is a key factor in Shiga toxin (Stx)-producing E. coli, a common pathogen involved in foodborne disease outbreaks. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) based approach has been used in this study to identify commonly reported E. coli toxins, with a focus on Shiga toxins (Stxs). Different sample preparation methods using variable culture conditions and concentrations of mitomycin C (MMC), a common antibiotic/chemotherapy agent capable of stimulating Stx production, were first tested on reference strains EDL933 and 90-2380 by LC-MS/MS detection of tryptic digests of receptor-analogue affinity binding enriched Stx preparations from culture supernatants and lysates. A curated E. coli protein toxin database was also used for faster and more straightforward toxin identification. With eight more genetically confirmed E. coli strains examined to verify the method, this preliminary study indicates that receptor-analogue based affinity enrichment on cell lysate or supernatant is a sensitive and accurate method for Stx identification. Biological significance: The existence of Stx is very important for identifying Stx-producing E. coli and implementing a clinical treatment regime. This study demonstrates for the first time that using a curated E. coli toxin database, together with receptor-analogue-based affinity enrichment of Stxs after MMC treatment of E. coli, is an easy and appropriate approach for fast and accurate Stx identification through LC-MS/MS. Crown Copyright (C) 2017 Published by Elsevier B.V. All rights reserved.

人类肠道病毒(enteroviruses ,EV)是小RNA病毒科的一个属 ,目前有100多种血清型[1-2] .EV主要包括脊髓灰质炎病毒、柯萨奇病毒(coxsackievirus ,CV )、Echo病毒和新型EV ,可导致手足口病 (hand ,foot ,and mouth disease , HFMD)、无菌性脑膜炎、脑炎、脊髓灰质炎、疱疹性咽峡炎等多种疾病[2] .柯萨奇病毒 A 组 4 型(CV-A4 )是能引起HFMD的A组EV的一种 ,也是疱疹性咽峡炎的重要病原体[3-13 ] .本研究对2016年郑州市一例疱疹性咽峡炎患儿进行了临床症状的分析和病原学分子鉴定 ,同时对鉴定的病原样本的基因组进行了测定和分析.

Objective To analyze the VP1 sequences of coxsackievirus A16(CA16) causing neu-rologic complications. Methods Clinical samples and epidemiological information were collected from pa-tients with viral encephalitis. Coxsackievirus A16 in these samples were first detected with real time RT-PCR and then isolated. RT-PCR was performed to amplify VP1 sequences and the amplified products were se-quenced. DNAStar 5.0 and Mega 5 were used for sequence analysis. All data was analyzed with SPSS statis-tical software. Results Fifteen samples were collected from 12 patients with hand, foot and mouth disease (HFMD) complicated by neurologic complications. Eight patients had the symptoms of fever, skin rash, signs of meningeal irritation and neck rigidity. No typical cluster was associated with clinical features or the time of onset. Both pharyngeal/anal swab and serum samples were collected from three patients (patient′s number:01111,01169 and 01130). The two samples collected from both 01111 and 01130 patients shared 100% similarity in nucleotide and amino acid based on VP1 sequences,while those from 01169 patient dif-fered in only one base. The 15 CA16 isolates were highly similar in VP1 gene, sharing 94.5%-100% ho-mology in nucleotide sequences and 98.0%-100% homology in amino acid sequences. These 15 isolates showed 68.5%-70.5% identities in nucleotide sequences and 90.5%-91.9% identities in amino acid se-quences with the CA16 prototype strain G10. Phylogenetic analysis revealed that based upon VP1 sequences, all of the 15 CA16 isolates grouped into genotype B subtype 1b (B1b), which was further classified into three clusters. Conclusion All of the 15 CA16 isolates causing neurologic complications belonged to B1b sub-genotype. Understanding the molecular epidemiology of CA16 would be essential for controlling morbidi-ty rates of HFMD and vaccine research.

目的 综合处置一起一犬伤13人的狂犬病暴露事件,评价预防处置措施效果,为规范化处置一犬伤多人事件提供依据.方法 根据《狂犬病暴露后预防处置工作规范(2009版)》对一犬伤13人事件中的13人进行伤口清洗、狂犬病疫苗和/或狂犬病免疫球蛋白接种;分别采用RT-PCR方法和RFFIT方法进行伤人犬的实验室诊断和狂犬病暴露后处置人群免疫效果评价.结果 伤人犬被打死、焚烧后深埋处理.犬脑组织标本经RT-PCR检测狂犬病病毒N、L基因均有特异性条带出现.13例伤者均为Ⅲ级暴露,均在24h内主动至当地狂犬病预防处置门诊进行暴露后预防处置,其全程免疫后第15d的血清抗狂犬病病毒中和抗体水平均＞0.5 IU/mL.结论 伤人犬为狂犬病病犬,13例狂犬病暴露后免疫人群全程免疫后血清中和抗体均达到保护性水平.及时、有效的多部门合作对一犬伤多人事件的处置具有积极意义.

Background: Rabies is a serious reemerging zoonosis in China. At present human rabies cases are primarily diagnosed based on clinical presentation.Case presentation: In August 2012, a woman and her son were attacked by a stray dog in Henan, China. The son received rabies postexposure prophylaxis (wound treatment followed by vaccine, no immunoglobulin), however, the mother did not. Rabies infection was subsequently laboratory confirmed in the mother and she died in December; her son is alive and healthy after 2 years of follow-up.Conclusion: This report documents that the timely utilization of postexposure prophylaxis is a required measure in preventing rabies after exposure to an animal bite.

目的 通过对河南省濮阳县某农村一起人间狂犬病散发疫情进行现场调查与处置,总结提出规范的调查与处置工作流程.方法 对河南省濮阳县报告的一起人间狂犬病散发疫情进行现场调查与处置,分别进行流行病学调查、现场消杀、防控知识宣传、工作总结评估,同时开展肇事犬追踪和疑似病例搜索.最后提出人间狂犬病疫情综合防控措施,及对疫情处置效果进行评估.结果 经现场核实,报告病例为临床诊断狂犬病病例.成立了疫情调查处置队伍,分流行病学调查2人,现场消杀1人,防控知识宣传1人,工作总结评估1人.列出疫情处置所需物品清单,对疫点及有关物品、用具进行消毒,无一犬伤多人情况,未搜索到疑似病例,采取发放宣传单、村内广播等形式进行了狂犬病防控知识宣传.经狂犬病常见潜伏期2～3个月后,无新发病例出现.结论 狂犬病等人兽共患病疫情的规范处置应包括响应准备、现场处置、总结评估3个阶段.

目的 总结和分析河南省人间狂犬病流行特征,探讨防控策略.方法 收集并整理河南省1951-2015年人间狂犬病疫情相关资料,对2007-2015年病例进行流行病学调查,用SPSS17.0建立数据库并进行统计分析.结果 河南省人间狂犬病上世纪80年代曾严重流行、90年代得到有效控制.本世纪初以来疫情上升明显,近10年又有下降趋势.病例以农村地区居民为主,男多于女,多为35-65岁组及15岁以下组人群.7-9月份是发病高峰期.豫东和豫南是人间狂犬病病例较为集中区域.病例平均潜伏期为60 d,平均病程为3d.病例暴露后约45.9％未作任何伤口处理,76.5％oo未曾接种疫苗,伤口肉眼可见出血的占93.6％,抗狂犬病免疫球蛋白注射率仅为0.64％.传染源98.7％是犬只,其中非栓养犬占82.1％以上.犬只狂犬病毒感染率为0.63％～6.00％.农村地区犬只免疫率为0.64％oo,人犬比值平均为6.50.结论 河南省是狂犬病流行地区,应开展以农村地区为重点的综合防控.

目的 对一例暴露因素不明的人狂犬病病例进行实验室确诊,通过分子流行病学分析探索其可能的感染来源.方法 通过对存活疑似人狂犬病病例的唾液、脑脊液、血清标本采用直接免疫荧光试验、反转录-聚合酶链式反应和快速荧光灶抑制试验进行实验室确诊;通过时空进化分析探索该病例可能的感染来源.结果 病例唾液标本直接免疫荧光试验结果呈阳性,病例血清标本经快速荧光灶抑制试验检测抗狂犬病病毒中和抗体呈阳性.病例唾液标本经反转录-聚合酶链式反应获得狂犬病病毒核蛋白基因预期扩增片段,序列测定进一步证实为狂犬病病毒.该狂犬病病毒株与2011年安徽省一株犬源狂犬病病毒株核苷酸序列同源性最高,为98.4％.结论 该疑似狂犬病病例可确诊为狂犬病病例.其发病源于至少2011年以来的某次不自觉的狂犬病病毒感染.