Coverage of enterovirus 71 vaccine among children born between 2018 and 2021 in Ningbo City / 预防医学

Objective@# To investigate the coverage of enterovirus 71 (EV71) vaccine among children born between 2018 and 2021 in Ningbo City, Zhejiang Province, so as to provide insights into formulating EV71 vaccination strategy.@*Methods@#Data pertaining to demography and EV71 vaccination among children born between 2018 and 2021 in Ningbo City were obtained from the Zhejiang Immunization Information Management System, and the coverage of vaccination, full-dose vaccination and timely vaccination of EV71 vaccine were analyzed. @*Results@#Totally 416 176 children were born between 2018 and 2021, including 216 977 boys (52.14%) and 199 199 girls (47.86%). The coverage rates of vaccination, full-dose vaccination and timely vaccination was 61.50%, 58.70% and 32.53%, respectively. The coverage rates of vaccination, full-dose vaccination and timely vaccination among local children (71.43%, 70.04% and 40.24%) were higher than those among migrant children (54.05%, 50.18% and 26.73%; P<0.05). The coverage rates of vaccination, full-dose vaccination and timely vaccination among urban children (67.72%, 65.06% and 40.37%) were higher than those among rural children (54.11%, 51.14% and 23.21%; P<0.05). The coverage rates of vaccination were 54.98%, 61.24%, 65.10% and 66.72%, the coverage rates of full-dose vaccination were 52.16%, 58.72%, 62.44% and 63.39%, and the coverage rates of timely vaccination were 22.47%, 25.86%, 39.14% and 48.03% by the born year from 2018 to 2021, all showing increasing trends year by year (P<0.05). @*Conclusion@#The coverage of vaccination, full-dose vaccination and timely vaccination of EV71 vaccine appears a tendency towards a rise among the 2018-2021 birth cohorts in Ningbo City, while the coverage of timely vaccination need to be improved. Health education for EV71 vaccination should be enhanced for rural and migrant children.

Absorption characteristics of aluminum hydroxide adjuvant with inactivated enterovirus 71 / 中国生物制品学杂志

@#Objective To explore the absorption characteristics of aluminum hydroxide adjuvant with inactivated enterovirus 71(EV71).Methods The morphology,purity and particle size distribution of inactivated EV71 particles were analyzed by transmission electron microscope,size exclusion chromatography HPLC(SEC-HPLC) and dynamic light scatter(DLS),and the morphology of aluminum hydroxide adjuvant nanoparticles was observed by transmission electron microscope.Using inactivated EV71 antigen content(4 000,6 000,8 000,10 000,11 000,12 000,13 000,14 000,20 000,30 000 U/mL),aluminum hydroxide adjuvant concentration [0.35,0.25,0.17,0.085 mg/mL(aluminum content)],adsorption time(0.30 and 120 min),ionic strength(sodium chloride concentration of 0.15,0.75 and 1.25 mol/L)and phosphorus-aluminum molar ratio(P/Al,0.15,0.64,2.08 and 7.87) as variables,the adsorption characteristics of aluminum hydroxide adjuvant with inactivated EV71 antigen were investigated.Results Inactivated EV71 particles mainly existed in the form of intact virus particles with a diameter of about 30 nm;Aluminum hydroxide adjuvant showed the characteristics of nanocrystallization,and the particle size was distributed within 200～700 nm.The inactivated EV71 antigen at the concentration of no more than 11 000 U/mL was completely absorbed by aluminum hydroxide adjuvant of 0.35 mg/mL(aluminum content),and no free antigen was detected in the supernatant;while from 12 000 U/mL,the content of free antigen in the supernatant increased with the increase of antigen content;the inactivated EV71 antigen of 250 U/mL was completely absorbed by various concentrations of aluminum hydroxide adjuvant,and no free antigen was detected in the supernatant;the adsorption effect of aluminum hydroxide adjuvant consistent after incubation for different time,and no free antigen was detected in the supernatant.Under the conditions of single dose of vaccine with aluminum hydroxide content(0.35 mg/mL) and inactivated EV71 antigen content(250 U/mL),sodium chloride ion strength had no effect on the adsorption of inactivated EV71 virus,while phosphate ion concentration significantly effected the adsorption.Conclusion Aluminum hydroxide adjuvant in a single dose of vaccine completely absorbed inactivated EV71 antigen,and group replacement played an important role.

Epidemiological features of hand, foot, and mouth disease among children under 6 years in Wuxing District / 预防医学

Objective@# To investigate the epidemiological characteristics of hand, foot, and-mouth disease (HFMD) among children under 6 years in Wuxing District, Huzhou City from 2013 to 2021, so as to provide insights into HFMC control among children. @* Methods @#The surveillance on HFMD and enterovirus 71 (EV71) vaccination among children under 6 years in Wuxing District, Huzhou City from 2013 to 2021 were captured from China Disease Prevention and Control Information System, and the temporal, human and spatial distributions of HFMD cases and pathogenic detection results were descriptively analyzed.@*Results @#A total of 14 272 HFMD cases under 6 years were reported in Wuxing District from 2013 to 2021, with an average annual incidence rate of 465.75/104. There were 8 313 male children with HFMD (522.39/104 incidence) and 5 959 female children with HFMD (404.55/104 incidence), and higher incidence of HFMD was seen among male children than among female children (χ2=239.228, P<0.001). Most HFMD cases were found among children at ages of 1 to 3 years (10 006 cases, 70.11%), and the incidence of HFMD peaked during the period between May and July and between October and December each year. High incidence of HFMD was seen in Zhili Township (741.50/104), Yangjiabu Street (546.99/104) and Balidian Township (438.68/104). There were 717 laboratory-confirmed HFMD cases (5.02%), including 106 EV71-positive cases (14.78%) and 177 CoxA16-positive cases (24.69%). A total of 56 288 doses of EV71 vaccines were administered in Wuxing District from 2017 to 2021, with an annual average full-dose vaccination rate of 14.87%. The detection of EV71 infection was significantly lower after EV71 vaccination than before vaccination (4.49% vs. 24.93%; χ2=62.932, P<0.001).@* Conclusions @#The incidence of HFMD peaks in summer and winter among children under 6 years in Wuxing District, which predominantly occurs among children at ages of 1 to 3 years. The incidence of HFMD is high in Zhili Township, Yangjiabu Street and Balidian Township, and the number of EV71-positive cases significantly reduces after EV71 vaccination than before vaccination.

Epidemiological characteristics and etiological changes of hand-foot-mouth disease in Shijiazhuang City in 2009-2021 / 公共卫生与预防医学

Objective To understand the epidemiological and etiological characteristics of hand-foot-mouth disease (HFMD) in Shijiazhuang City from 2009 to 2021, and to provide scientific evidence for the prevention and control of HFMD. Methods The epidemic data and etiological data of HFMD in Shijiazhuang City from 2009 to 2021 were collected, and descriptive epidemiological methods were used for data analysis. Results The reported incidence of HFMD in Shijiazhuang showed a fluctuating downward trend from 2009 to 2021, with a high incidence every other year in most years. The proportion of severe cases and the mortality rate showed a decreasing trend (χ2severe cases=282.09, P2mortality=51.33, P2=4 507.84，Pr=-2.85，P22 =521.86，P2proportion of severe cases=93.71，P=0.000，χ2mortality rate=26.62，P=0.000，χ2proportion of EV71=1060.86，P=0.000). Conclusion The incidence of HFMD in Shijiazhuang presented a declining trend, and the dominant etiological changes of different cases were different. It is necessary to continue to strengthen the etiological monitoring, health education and EV71 vaccination for the prevention and control of HFMD.

Coverage of inactivated enterovirus 71 vaccine and analysis of epidemiological and etiological changes of hand-foot-mouth disease before and after vaccination in Jinshan District of Shanghai / 中国生物制品学杂志

@#ObjectiveTo analyse the coverage of inactivated enterovirus 71(EV71)vaccine and the impact on hand-foodmouth disease(HFMD)epidemiological and etiological changes in Jinshan District,Shanghai,and provide evidence for improving the prevention and control strategy of HFMD in this area.MethodsThe vaccination data of inactivated EV71vaccine from 2016 to 2019 was collected in Jinshan Immunization Information Management System of Shanghai to describe the vaccination characteristics;The data of HFMD cases in Jinshan District from 2013 to 2019 were extracted from Chinese Disease Prevention and Control Information System,and the surveillance etiological information of HFMD in the same period was obtained,which was compared for the differences of HFMD incidence and pathogen positive detection rate before and after vaccination.ResultsFrom November 2016 to December 2019,a total of 63 521 doses of inactivated EV71vaccine were applied in Jinshan District,with the first shot coverage of 22.57%,the full shot coverage of 21.05% and the two-dose completion coverage of 94.65%.There were significant differences in coverage between different years,months,current addresses,age groups and registers(P < 0.05).The highest coverage of first short was in 2018(33.45%),while full short in 2017(30.78%).More doses were applied during May to August,with highest coverage in 6 to 11 months old group and most doses in 1 year old group.The coverage of children in this city was higher than that of migrant children.There was no significant difference in the incidence of HFMD before and after vaccination(χ2= 0.427,P =0.513 ),while the incidence of severe disease,the positive detection rate of EV71 and the estimated incidence of HFMD infected with EV71 decreased significantly after vaccination(χ2= 15.312,41.431 and 432.342 respectively,each P <0.001).ConclusionVaccination with inactivated EV71 vaccine reduced the occurrence of HFMD EV71 infection and severe disease in Jinshan District,while the coverage was low,so it was necessary to pay attention to HFMD etiological changes to prevent other enterovirus infections.It is suggested to strengthen publicity and information technology to improve coverage,speed up the development of combined vaccine and provide more antibody protection.

Inhibitory effect of astragalus membranaceus injection on apoptosis and autophagy induced by enterovirus 71 / 中国生物制品学杂志

@#Objective To investigate the effect of astragalus membranaceus（AM）injection on apoptosis and autophagy of human gastric epithelial cell line（GES⁃1）induced by enterovirus 71（EV71）. Methods GES⁃1 cells were cultured in vitro and divided into infected group（EV71 infected at a MOI of 3 and control group（no virus infected）. The morpho⁃logical changes of EV71 infected cells were observed by inverted microscope. The level of VP1 in GES⁃1 cells infected with EV71 was detected by Western blot；CCK⁃8 assay was used to detect the viability of GES⁃1 cells infected with EV71；Nuclear staining with DAPI was used to observe the morphological changes of nuclear apoptosis infected with EV71. GES⁃1 cells were divided into control group（without virus infection），infection group and AM intervention group with final concentration of 1，2. 5，5 and 10 μg／mL，respectively. Western blot was used to detect the effect of AM intervention on the expression of apoptosis⁃related proteins Caspase⁃3，PARP and autophagy⁃related proteins LC3 and P62 in GES⁃1 cells infected withEV71. CCK⁃8 method was used to detect the effect of AM intervention on the viability of GES⁃1 cells infected with EV71. Results GES⁃1 cells were round，shrunken with nuclear pyknosis and uneven size；VP1 level increased（t = 41. 56，P < 0. 01），cell viability decreased（t = 19. 07，P < 0. 01），Caspase⁃3 and PARP proteins were cut off（t = 35. 29 and 3. 648， P < 0. 01 and 0. 021 8，respectively），LC3Ⅱ／LC3Ⅰ ratio increased（t = 10. 16，P = 0. 000 5）and P62 protein was degraded（t = 68. 68，P < 0. 01）；AM inhibited the degradation of Caspase⁃3，PARP and P62 proteins induced by EV71 （t = 52. 66，59. 60 and 40. 22，respectively，each P < 0. 01）and increased the ratio of LC3Ⅱ／LC3Ⅰ（t = 5. 521，P = 0. 005 3），andreducedtheinhibitoryeffectofEV71ontheviabilityofGES⁃1cells（t =4. 420，P =0. 0115）. Conclusion EV71 infection induced apoptosis of GES⁃ 1 cells and AM intervention inhibited EV71 induced apoptosis by inhibiting EV71 induced autophagy.

Epidemiological characteristics of hand, foot and mouth disease in Zhejiang Province from 2016 to 2019 / 预防医学

Objective@#To investigate the epidemiological characteristics of hand, foot and mouth disease ( HFMD ) in Zhejiang Province, so as to provide insights into HFMD control.@*Methods@#The incidence of HFMD in Zhejiang Province from 2016 to 2019 was collected from National Notifiable Disease Reporting System. The temporal distribution, human distribution, regional distribution and pathogenic typing of HFMD were descriptively analyzed in Zhejiang Province from 2016 to 2019.@*Results@#Totally 642 305 cases with HFMD were reported in Zhejiang Province from 2016 to 2019, including 121 severe cases and 9 fatal cases. The annual incidence of HFMD was 335.88/105, 147.76/105, 435.63/105 and 221.77/105, respectively. The incidence of HFMD peaked from May to July each year, and the three highest annual incidence included Ningbo, Jinhua and Wenzhou cities, while the lowest annual incidence was seen in Zhoushan City. The HFMD cases were predominantly found in children at ages of 1 to 5 years ( 537 738 cases, 83.72% ), and in children living at home ( 419 408 cases, 65.30% ). The average annual incidence of HFMD was higher in males than in females ( 328.23/105 vs. 239.99/105; P<0.05). The dominant pathogens gradually shifted from enterovirus 71 ( EV71 ) to Coxsachievirus A16 ( CA16 ), and other enteroviruses remained as the main pathogenic subtypes.@*Conclusions@#The incidence HFMD was high in summer and autumn in Zhejiang Province from 2016 to 2019. Children living at home are at a high risk of HFMD, and CA16 type gradually became the dominant pathogen of HFMD.

Effect of Pien Tze Huang against EV71 / 中国中药杂志

This study aims to investigate the inhibitory effect of Pien Tze Huang(PZH) on enterovirus 71(EV71). To be speci-fic, chemiluminescence method was adopted to evaluate the toxicity of PZH to African green monkey kidney(Vero) cells and human rhabdomyosarcoma(RD) cells, and cytopathic effect(CPE) method to assess the inhibition on EV71-GFP reporter virus and EV71 C4 wild-type virus. The results showed that PZH had low cytotoxicity to Vero cells and RD cells, with the half-maximal cytotoxic concentration(CC_(50)) of about 0.691 3-0.879 2 mg·mL~(-1) for the two. In addition, PZH can effectively inhibit the replication of EV71 within the non-cytotoxic concentration range, and dose-dependently alleviate the cytopathic changes caused by virus infection, with the half-maximal effective concentration(EC_(50)) of 0.009 2-0.106 3 mg·mL~(-1). On the basis of the above results, the green fluorescent protein(GFP), indirect immunofluorescence assay(IFA), and median tissue culture infective dose(TCID_(50)) were employed to assess and verify the anti-EV71-GFP and anti-EV71 C4 activity of PZH. The results demonstrated that PZH can dose-dependently lower the expression of GFP by EV71-GFP and structural protein VP-1 by EV71 C4 and decrease the production of progeny infectious viruses. The EC_(50) of PZH for EV71-GFP and EV71 C4 was about 0.006 0-0.006 2 mg·mL~(-1) and 0.006 6-0.025 6 mg·mL~(-1), respectively. This study suggested that PZH may exert antiviral activity by acting on EV71 and interfering with the expression of VP-1. At the moment, there is still a lack of specific anti-EV71 drugs. This study proposed a new idea for the symptomatic treatment of EV71 infections such as hand-foot-mouth disease and verified an effective drug for the treatment of EV71 infections.

Regulation of RDN on Th1/ILC1 cell imbalance in HFMD patients caused by EV71 infection / 中国天然药物

Enterovirus 71 (EV71) infection is more likely to cause hand, foot and mouth disease (HFMD) in children, which can lead to neurogenic complications and higher mortality. As a commonly used clinical medicine, Reduning injection (RDN) helps to shorten the symptoms of patients with HFMD and facilitate the early recovery of children. However, the regulatory mechanism of RDN on the HFMD immune system disorder caused by EV71 remains to be discussed. This study collected detailed treatment data of 56 children with HFMD who entered the affiliated Children's Hospital of Nanjing Medical University during 2019. Retrospective analysis of clinical data showed that the symptoms of the RDN treatment group were improved compared with the untreated group. To explore its mechanism, the relevant detection indicators were detected by flow cytometry, enzyme-linked immunosorbent assay and real-time quantitative PCR. It was found that the number and function of innate immune (ILCs) and adaptive immunity (Th1, Th2 and secreted cytokines) were reduced, suggesting that RDN plays a role by regulating cellular immunity. The in vitro differentiation inhibition test further confirmed that RDN affected Th1 differentiation by inhibiting the expression of transcription factors on the basis of Th1 cell differentiation in vitro.

Inactivated enterovirus 71 vaccination in Hannan District of Wuhan / 公共卫生与预防医学

Objectve To analyze the EV71 vaccination of children in Hannan District of Wuhan, providing a reference for the adjustment of EV71 vaccination strategy. Methods Based on the Hubei Province Immunization Program Information Management System, the general information and the Immunization information of the inactivated EV71 vaccine in children born between January 1, 2016 and December 31, 2019 managed by the inoculation department of Hannan was collected. Descriptive statistical analysis was used to calculate the vaccination rate and vaccination quality. Results Vaccination rate for first dose and full vaccination of EV71 vaccine of children in Hannan were 49.95% and 42.82% . Vaccination rate for first dose of local children was lower than that of ecdemic children (P <0.005), and Vaccination rate for full vaccination was slightly higher than that of ecdemic children (P=0.859). Vaccination rate for first dose and full vaccination of children in rural  areas were lower than those in urban areas (P <0.005) ). Vaccination rates were obvious differences among children of various age groups (P <0.005). Full vaccination rate of 6-11 months of age were 15.66%, full vaccination rate of children of the later the time of birth were the higher(P <0.005), full vaccination rate of the local children were lower than the ecdemic children (P <0.005), while which of children in urban areas were higher than children in rural areas (P <0.005). Between the two dose of vaccination, the unqualified vaccination rate was 5.54%, the timely vaccination rate was 66.94%, and the qualified vaccination rate was 27.52%. The timely vaccination rate of children of the later the time of birth were the higher (P <0.005). The unqualified vaccination and timely vaccination of local children were higher than ecdemic children (P = 0.02), Unqualified vaccination for children in urban areas is lower and timely vaccination is higher than that in rural areas (P <0.005). Conclusion The EV71 vaccination rate of children in Hannan increased year by year, but was far from establishing herd immunity barrier. Propaganda of HFMD prevention and control knowledge must be strengthened, increasing the coverage rate of full vaccination of children before 1 year of age and the timely vaccination rate of the second dose.

Enterovirus 71 can induce autophagy and apoptosis of THP-1 macrophages / 南方医科大学学报

OBJECTIVE@#To investigate enterovirus 71 (EV71)-induced of autophagy, apoptosis and the related signaling pathways in THP-1 macrophages.@*METHODS@#THP-1 macrophages were infected with EV71 at the multiplicity of infection (MOI) of 0.1 for 2, 8 or 16 h, and the cell proliferation and toxicity were analyzed using CCK-8 kit. The intracellular viral nucleic acid in THP-1 macrophages were detected by fluorescence quantitative PCR, and the ultrastructural changes of the cells were observed using transmission electron microscopy. Cell apoptosis induced by EV71 infection was detected using Hoechst 33342 staining and AnnexinV/PI double staining. Western blotting was performed for analysis of changes in autophagy and apoptosis of the cells and in the expressions of the related proteins. The effect of EV71 infection on apoptosis of THP-1 macrophages incubated with 3-MA and Ac-DEVD-CHO inhibitor for 2 h was assessed using Western blotting.@*RESULTS@#EV71 infection significantly lowered the cell survival rate of THP-1 macrophages at 2, 8 h and 16 h after the infection ( < 0.05). The total copy number of viral nucleic acid in THP-1 macrophages incubated with EV71 increased significantly and progressively over time ( < 0.01). Intracellular autophagosomes and virions could be seen in EV71-infected THP-1 macrophages. The total apoptotic rate of the infected cell also increased significantly over time ( < 0.01). EV71 infection significantly increased LC3 conversion (LC3-Ⅱ/ LC3-I) and the expression of cleaved caspase 3 protein and decreased the protein expressions of p62, Bcl-2 and caspase-3 ( < 0.01) without causing obvious changes in cleaved caspase-8 (>0.05). 3-MA significantly inhibited the EV71-induced autophagy of THP-1 macrophages and reduced LC3 conversion (LC3-Ⅱ/LC3-I) and p62 protein expression at 8 h after EV71 infection ( < 0.01). Compared with DMSO, Ac-DEVD-CHO significantly inhibited EV71-induced apoptosis of THP-1 macrophages (15.5% 7.7%, < 0.01).@*CONCLUSIONS@#EV71 not only can infect and replicate in THP-1 macrophages, but also induces autophagy and cell apoptosis possibly by activating LC3/p62 autophagy pathway and caspase apoptosis pathway.

New advances in the discovery of anti-enterovirus-71 agents / 药学学报

In recent years, enterovirus infection has become a frequent epidemic and developed into an important public health problem. For example, hand-foot-mouth disease has become a common infection among children in China. Hand-foot-mouth disease (HFMD) has been spreading globally since 1997, especially in the Asia-Pacific region. Enterovirus 71 (EV71) is one of the main pathogens causing HFMD. And now there is no drug available to treat EV71 infection. This review summarizes the research progress of anti-enterovirus-71 drugs from the perspective of medicinal chemistry.

Synthesis and synthetic process optimization of the enterovirus 71 3C protease inhibitor NK-1.8k / 国际药学研究杂志

Objective: To synthesize the enterovirus 71 3C protease inhibitor NK-1.8k and optimize the synthetic process. Methods: With N-Boc-L-glutamic acid dimethyl ester as the starting material, the target compound NK-1.8k was synthesized via the substitution, reductive amination, deprotection, amide condensation, hydrolysis, and reduction reactions. Compared with the original synthetic route, the tandem string type in the important intermediate 5 synthesis was changed to parallel type, thereby the total synthetic reactions were condensed from seven steps to six steps, and some post-processing methods were also optimized. Results: The structures of intermediates and the target compound were confirmed by MS and 1H NMR data, and the total yield of the target compound synthesis was increased to 13.3% from 10.7% of the original route. Conclusion: The synthetic route established in this article for NK-1.8k is reasonable and feasible, the raw materials used are cheap and easily available, the operation is simple, most of the reaction conditions are mild and controllable, the post-processing is simple, the intermediates are easy to separate and purify, the steps are short, and the yield is high. This method provides a valuable reference for the further synthesis of NK-1.8k and similar products.

VP1-binding protein glucose-regulated protein 78 as an important mediator for enterovirus 71 infecting human brain microvascular endothelial cells

Purpose: Enterovirus 71 (EV71) is one of the main pathogens causing hand, foot and mouth disease, which could even induce severe brain damage in some patients. As the underlying mechanism of the invasion and replication process still remains largely unknown, we investigated the role of candidate proteins expressed during EV71 invasion in human brain microvascular endothelial cells (HBMECs) to delineate the pathophysiological mechanism of EV-71 infection. Materials and Methods: Ninety-one candidate EV71-associated proteins which could bind the major capsid protein (viral protein 1 [VP1]) of EV71 on the HBMEC were identified by applying an analysis of glutathione-S-transferase pull-down coupling with liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS). Seventy-eight kDa glucose-regulated protein 78 (GRP78) binding to the VP1 protein was further validated by co-immunoprecipitation, immunofluorescence and western blot analysis. To explore the role of GRP78 in EV71 infection, GRP78 was knocked down and overexpressed in HBMEC and was verified by TCID50 assay. Results: LC-ESI-MS/MS-identified 91 proteins were subjected to gene ontology analysis, and on molecular and biological function analysis revealed GRP78 act as an important binding protein in mediating EV71 infection. In addition, immunofluorescence demonstrated the co-localisation of GRP78 and VP1 in cytoplasm of the infected HBMEC. The TCID50 assay showed that knockdown of GRP78 could attenuate the replication capacity of EV71 in HBMEC, and the overexpression could increase the virus titre in HBEMC at 24 h post-infection suggesting that GRP78 was associated with the replication capacity of EV71 in HBMEC. Conclusion: These findings provided evidence that GRP78 plays an important role during the progression of EV71 infection as a mediator in HBMEC.

Surveillance results of adverse events following immunization of enterovirus 71 inactvated vaccine / 预防医学

Objective@#To evaluate the safety of enterovirus 71（EV71）inactivated vaccines produced by Wuhan Institute of Bio Products through learning the incidence of adverse events following immunization（AEFI）in children aged 6 months to 3 years old after vaccination.@*Methods@#According to the national requirement for vaccine safety monitoring program，data of AEFI cases after inoculated EV71 vaccines from September 2017 to October 2018 in Zhejiang Province was collected by combining active and passive monitoring. Demographic characteristics，space distribution，dose and incidence of AEFI were analyzed.@*Results@#A total of 107 503 children were included in this study，of which 27 173 were actively monitored and 80 330 were passively monitored. The monitoring results showed that 288 cases of AEFI occurred after inoculated EV71 vaccines，the reported incidence rate was 267.90/100 000. Of 288 cases，266 cases had common reactions（247.43/100 000），14 cases had abnormal reactions（13.02/100 000）and 8 cases had complications（7.44/100 000）；145 cases were passively monitored（180.51/100 000）and 143 cases were actively monitored（526.26/100 000）；150 cases were males and 138 cases were females，with the males to females ratio of 1.09∶1. The AEFI of EV71 vaccines were reported in all the cities of Zhejiang Province，with Jinhua，Taizhou and Hangzhou ranking the top three and accounting for 50.69% of all the reported cases. About 202 cases of AEFI（70.14%）occurred after the first dose of EV71 vaccines，and 163 cases（56.60%）occurred on the day of inoculation.Most of common reactions were fever，swelling and sclerosis. The harm of the reported abnormal reactions was mild，with 8 cases of anaphylactic rash and no other serious abnormal reactions were found. @*Conclusion@#The reported incidence rate of AEFI of the domestic EV71 vaccine in children aged from 6 months to 3 years is 267.90/100 000，without serious abnormal reactions，suggesting that the EV71 vaccine is safe.

Influencing factors for enterovirus 71 vaccination among children aged below 3 years in Xicheng District / 预防医学

Objective@#To understand the situation of inoculation and influencing factors of children in the Xicheng District of Beijing after the enterovirus 71（EV71）vaccine was marketed .@*Methods@#A questionnaire survey was conducted among the parents of children aged 6 to 35 months in the vaccination clinics in Xicheng District from May to September 2017. Demographic characteristics,awareness of hand-foot-mouth disease and EV71 vaccine,the access to these knowledge,acceptance of the vaccine price and advice on the management were collected. The influencing factors for EV71 vaccination were analyzed by multivariate logistic regression .@*Results@# There were 1 850 out of 1 885 parents investigated,with the effective rate of 98.14%. The EV71 vaccination rate of children aged 6 to 35 months was 27.03%. The awareness rate of hand-foot-mouth disease and EV71 vaccine in parents was 55.95%. The results of logistic regression analysis showed that the parents with the annual household income of 50 000 yuan and above （OR50 000 yuan- =2.665,95%CI：1.535-4.628;OR100 000 yuan-=4.732,95%CI：2.830-7.914;OR200 000 yuan-=3.576,95%CI：2.084-6.135）,with acceptance of 100 yuan and above in the price of the vaccine （OR100 yuan-= 9.487,95%CI：6.873-13.096;OR200 yuan-=5.940,95%CI：3.465-10.183;OR300-500 yuan-=2.415,95%CI：1.183-4.933）,with more than two sources of the information for EV71 vaccine （OR=3.062,95%CI：2.306-4.065）,without the care about the management of the EV71 vaccine（OR=1.650,95%CI：1.156-2.356）were more likely to have their children vaccinated;while the parents who lived in bungalows and cabinets （OR=0.589,95%CI：0.386-0.899）,who were medical staff（OR=0.240,95%CI：0.118-0.487）,who were not residents of Beijing（OR=0.587,95%CI：0.399-0.863）,who had more than one children （OR=0.338,95%CI：0.236-0.483） were less likely to have their children vaccinated .@*Conclusion@#The parents of children aged below three years in Xicheng District of Beijing should improve their awareness of hand-foot-mouth disease and EV71 vaccine. The vaccination rate of EV71 were mainly related to the registration,occupation,number of children, household income,the access to the information about vaccine,acceptance of the vaccine price and advice on the management in parents.

Post-marketing observation on safety of inactivated enterovirus A71 vaccine (human diploid cell) / 中华预防医学杂志

Objective@#To evaluate the post-marketing safety of inactivated Enterovirus type 71 (EV-A71) vaccine (human diploid cell) .@*Methods@#A total of 20 191 healthy children aged 6 to 59 months were invited to receive 2 doses of EV-A71 vaccine in Zhejiang Province from September 2016 to December 2017. Child caregivers were followed up on the 4th or 5th day after each EV-A71 vaccination, and the incidence of local, systemic, and other adverse events within 3 days after vaccination was recorded to assess vaccine safety. Describe the differences in adverse events among children with different characteristics.@*Results@#A total of 32 230 doses were observed in this study, of which 20 191 and 12 039 were vaccinated for the first and the second dose, respectively; and the incidence of adverse events within 3 days was 2.045% (413 doses) and 1.611% (194 doses), respectively. After the first and the second dose, the number of systemic adverse events was the highest, 371 and 175 cases, respectively, with an incidence of 1.837% and 1.454%, respectively; the number of local adverse events was the lowest, 14 and 2 doses, respectively, with an incidence of 0.069% and 0.017%. Local adverse events occurred after vaccination were generally mild, and only 2 patients had level of 3; among the systemic adverse events, 39 patients had a fever level of 3 or higher, accounting for 8.2% of the total fever. Most of the symptoms in the local adverse events did not require treatment, only 3 cases of vaccination site rash and 2 cases of pruritus were self-purchased drugs or outpatient treatment; except for 5 cases of fever, the other symptoms were not hospitalized in the case of systemic adverse events.@*Conclusion@#The incidence of adverse events within 3 days after vaccination with EV-A71 vaccine was low in children, mainly systemic adverse events, and the prognosis was good.

Post-marketing multi-center safety surveillance of inactivated enterovirus A71 vaccine (Vero cell) / 中华预防医学杂志

Objective@#To evaluate the post-marketing safety profiles of the inactivated enterovirus type 71 (EV-A71) vaccine (Vero cell) after routine inoculation.@*Methods@#Eleven cities of Zhejiang Province, Fengtai district of Beijing, Qinnan district, two counties as Pingle and Pingguo of Guangxi Zhuang Autonomous Region, and Dongtai city of Jiangsu Province were selected as the field sites. A total of 45 239 subjects were enrolled in this study from children who seeked the vaccination of EV-A71 vaccine during the period from July, 2016 to June, 2018. Different sampling method were adopted in different sites. All vaccinated children were invited to participate in the study in Fengtai and Dongtai, however, systematic sampling method were adopted in other sites. Active surveillance was conducted and information about adverse reactions (ARs) occurred in 30 min, 3 d and 30 d following each dose of EV-A71 immunization was collected by field observation, phone-call or face-to-face interview. The incidence of ARs in different types, symptoms and grades were described.@*Results@#In total, there were 45 239 children who received 71 243 doses EV-A71 vaccine. The overall incidence of ARs was 1.079% (769 doses), with the highest incidence of 1.182% (177/14 973) in 5-11 month group and the lowest incidence of 0.849% (18/2 119) in ≥ 36 month group among different age groups. There was a higher incidence in solicited ARs, which was 1.047% (746 doses). The incidences of grade 1 and grade 2 ARs were also higher, which were 0.404% (288 doses) and 0.554% (395 doses), respectively. No grade 4 ARs occurred. The doses of the first and the second vaccination was 40 736 and 30 507, respectively, and the incidences of ARs were 1.281% (522 doses) and 0.810% (247 doses). Also, the incidences of ARs were 0.091% (37 doses) and 0.043% (13 doses) in local, and 1.168% (476 doses) and 0.760% (232 doses) in system. The symptoms of ARs after the two doses of vaccination were basically the same. Redness at the injection site was the most common local ARs after each dose vaccination, with doses of 24 and 11, while fever was the most common systemic ARs, with doses of 362 and 190. Moreover, ARs mainly occurred in 30 min to 3 d after each dose vaccination, with incidence of 1.016% (414 doses) and 0.698% (213 doses) in the first and second dose, respectively.@*Conclusion@#The ARs had a low incidence after vaccination in children and most were mild or moderate. EV-A71 vaccine with good safety is suitable for inoculation in a large scale.

EV71 induces incomplete cellular autophagy to promote self-replication / 中华实验和临床病毒学杂志

Objective@#To investigate the relationship between enterovirus 71 (EV71) and autophagy and the effect of autophagy on viral replication.@*Methods@#RD cells were pretreated with rapamycin, Chloroquine (CQ) and 3-methyladenine (3-MA). The effects of different stages of autophagy on viral replication were detected by Western blotting and plaque assay.@*Results@#The study showed that EV71 infection could induce incomplete autophagy in RD cells. Replication of EV71 was promoted in the CQ treatment group, while the level of replication of the rapamycin and 3-MA treatment groups was reduced. Further studies have shown that mTOR is a key molecule affecting EV71 replication, and inhibition of mTOR by rapamycin can inhibit the synthesis of viral RNA.@*Conclusions@#The effects of different stages of autophagy on viral replication are also different. Inhibition of autophagic lysosome degradation promotes viral replication, while inhibition of the early stage of autophagy or promotion of autophagy reduces the level of viral replication. mTOR can affect the replication of EV71 at RNA level.

Sequence analysis of enterovirus 71 3′UTR and construction of a recombinant cDNA infectious clone / 中华实验和临床病毒学杂志

Objective@#To sequence the 3′UTR of enterovirus 71 strains, investigate its foundation and impact in virulence by constructing a 3′UTR-replaced recombinant cDNA infectious clone.@*Methods@#Viral RNA of EV-A71 isolated viruses were extracted, and the nucleotide analysis was performed after sequencing. The 3′UTR of a full-length infectious clone of SDLY107 strain was replaced by its corresponding part of SDLY1 strain, and then the recombinant virus was constructed and identified.@*Results@#The nine isolated strains were classified into sub-genotype C4a of enterovirus (EV)-A71 by analysis, and nucleotide sequence homology for 3′UTR were 94%-100%. 3′UTR of EV-A71 strains may be associated with its pathogenicity. Identification of the rescued virus by sequencing and indirect immunofluorescence confirmed the successful construction of infectious recombinant virus.@*Conclusions@#Sequence analysis indicated that the 3′UTR may be involved in the pathogenicity of EV-A71. The recombinant virus SDLY107(1-3′UTR) was rescued successfully. Our study may provide evidence for further research on the influence of 3′UTR on the virulence of enterovirus 71.