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Homocysteine is closely associated with extracranial and intracranial atherosclerosis, and its main pathogenesis includes oxidative stress, lipid metabolism disorder and vascular endothelial dysfunction. As a protein modification related to homocysteine, homocysteinylation can promote the occurrence and development of cerebral atherosclerosis by promoting oxidation, changing lipid function and destroying vascular endothelial function. This article reviews the role of homocysteinylation in cerebral atherosclerosis, and discusses the possibility of preventing cerebral atherosclerosis by homocysteinylation.
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Intracranial atherosclerotic stenosis (ICAS) is the main cause of ischemic stroke. Endovascular therapy (EVT) is a method of treating symptomatic ICAS, and in-stent restenosis (ISR) is an important factor affecting the efficacy of EVT. This article summarizes the influencing factors of ISR in patients with ICAS receiving EVT treatment.
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Objective@#To investigate the willingness to receive measles-containing vaccine (MCV) and its influencing factors among healthcare workers in the Yangtze River Delta region, so as to provide the evidence for improving the measles-containing vaccination rate@*Methods@#Healthcare workers were sampled from 19 medical institutions in each of Shanghai Municipality, Jiangsu Province, Zhejiang Province and Anhui Province for questionnaire surveys using a multi-stage stratified convenience sampling methods from July 2020 to March 2021. Participants' gender, age, educational level, professional title, measles-containing vaccination, awareness of MCV and willingness to receive MCV were collected, and the factors affecting the willingness to receive MCV were identified among healthcare workers using a multivariable logistic regression model. @*Results@#Totally 1 403 questionnaires were allocated, and 1 394 valid questionnaires were recovered, with an effective recovery rate of 99.36%. The respondents included 327 men and 1 067 women, with a male to female ratio of 1∶3.26, and 64.35% (897) were at ages of 31 to 50 years. There were 1 005 respondents with a bachelor degree (72.09%), 765 with middle and senior professional titles (54.88%), 676 with a history of measles-containing vaccination (48.49%), 1 176 with willingness to receive MCV (84.36%) and 218 without willingness to receive MCV due to convenience of vaccination (30.73%) and cost (19.27%). Multivariable logistic regression analysis showed that region (Zhejiang, OR=1.613, 95%CI: 1.054-2.470; Anhui, OR=2.058, 95%CI: 1.259-3.363), and no history of measles (OR=2.219, 95%CI: 1.302-3.781) were factors improving the willingness to receive MCV among healthcare workers, and hospital level (secondary, OR=0.483, 95%CI: 0.306-0.763; tertiary, OR=0.251, 95%CI: 0.160-0.394), history of measles-containing vaccination (no, OR=0.262, 95%CI: 0.172-0.399; unknown, OR=0.386, 95%CI: 0.266-0.559), and unawareness of MCV knowledge (OR=0.208, 95%CI: 0.081-0.536) were factors inhibiting the willingness to receive MCV among healthcare workers.@*Conclusions@#The willingness to receive MCV correlates with region, history of measles, hospital level, history of measles-containing vaccination and awareness of MCV knowledge among healthcare workers in the Yangtze River Delta region.
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Estimating the actual real-world effectiveness of the vaccine is an essential part of the post-marketing evaluation. This regression discontinuity design (RDD) using observational data is designed to quantify the effect of an intervention when eligibility for the intervention is based on a defined cutoff as age, making it suited to estimate vaccine effects. This approach can avoid the high cost and ethical issues; overcome difficulties in the organization and practice process in randomized controlled trials, which leads to a higher level of causal inference evidence and more realistic results. Here, we describe key features of RDD in general, and then specific scenarios, with examples, to illustrate that RDD are an essential tool for advancing our understanding of vaccine effects.
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Humans , Causality , Vaccine Efficacy , VaccinesABSTRACT
Japanese encephalitis (JE) virus is the leading cause of vaccine-preventable encephalitis in Asia and the Western Pacific, which mainly invades central nervous system. Vaccination is the most important strategy to prevent JE. Currently, both live attenuated Japanese encephalitis vaccines (JE-L) and inactivated vaccines (JE-I) are in use. Due to the supply of vaccines and the personal choice of recipients, there will be a demand for interchangeable immunization of these two vaccines. However, relevant research is limited. By reviewing domestic and foreign research evidence, this article summarizes the current situation of the interchangeable use of JE-L and JE-I, and makes recommendations when the interchangeable immunization is in urgent need, so as to provide reference for practical vaccination and policymaking in China.
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Humans , Encephalitis Virus, Japanese , Encephalitis, Japanese/prevention & control , Immunization , Japanese Encephalitis Vaccines , Vaccination , Vaccines, InactivatedABSTRACT
Objective @#To identify spatio-temporal distribution patterns of mumps in Zhejiang Province from 2008 to 2019, so as to provide insights into precise control of mumps.@*Methods @#The epidemiological data pertaining to mumps in Zhejiang Province from 2008 to 2019 were collected from the China Information System for Disease Control and Prevention. The time- and region-specific incidence of mumps was descriptively analyzed, and the spatio-temporal clustering characteristics of mumps incidence were identified using global and local spatial autocorrelation analyses and spatio-temporal scan analysis.@*Results @#A total of 167 196 cases of mumps were reported in Zhejiang Province from 2008 to 2019, and the incidence of mumps reduced from 56.08/105 in 2008 to 11.35/105 in 2019, with a reduction of 79.76%. There was no remarkable mumps seasonality since 2017; however, spatial autocorrelation was seen in mumps incidence in 2008, 2011 and 2012, respectively ( Moran's I coefficient=0.16, 0.18, 0.16; all P<0.05 ). Spatio-temporal scan analysis identified five spatio-temporal clusters of mumps incidence in 2008, 2011 and 2012, and the most likely spatio-temporal cluster was mainly concentrated in Cangnan County of Wenzhou City, covering 17 counties clustered in the time frame from 15 January 2008 to 28 February 2009, while the other four clusters were concentrated in Pinghu City of Jiaxing City, Kaihua County of Quzhou City, Fuyang District of Hangzhou City and Zhuji City of Shaoxing City, clustered in the time frame from 15 January 2008 to 30 June 2012. @*Conclusion @#The incidence of mumps appeared a tendency towards a reduction in Zhejiang Province from 2008 to 2019, and rebounded in some regions. No remarkable mumps seasonality was seen since 2017. The spatio-temporal clusters of mumps incidence were concentrated in Wenzhou and Jiaxing cities of Zhejiang Province in 2008, 2011 and 2012; however, no spatial cluster was identified since 2012.
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Objective:To investigate the efficacy and safety of endovascular treatment (EVT) in patients with acute ischemic stroke due to acute primary medium vessel occlusion (MeVO).Methods:Patients with primary MeVO received EVT in the stroke center of Jining First People's Hospital from January 2019 to January 2022 were enrolled retrospectively. MeVO was defined as occlusion of M2/M3 segment of middle cerebral artery, A2/A3 segment of anterior cerebral artery and P2/P3 segment of posterior cerebral artery. The symptoms, vascular recanalization and surgical complications were documented. The National Institutes of Health Stroke Scale (NIHSS) score at discharge and the modified Rankin Scale (mRS) score at 90 d after onset were followed up.Results:A total of 17 patients with primary MeVO were included, among them 16 successfully achieved vascular recanalization, 1 had distal emboli escape, and 1 had asymptomatic intracranial hemorrhage. There was significant difference in NIHSS scores before and after thrombectomy ( P<0.01). Ten patients (62.5%) had good functional outcome (mRS score 0-2), and 1 died of secondary pulmonary infection at 20 d after procedure. Conclusion:After strict risk/benefit evaluation and screening, EVT of acute ischemic stroke due to primary MeVO was technically feasible, and had good efficacy and safety.
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Intracranial atherosclerotic stenosis (ICAS) is one of the most common causes of ischemic cerebrovascular disease. Compared with other causes such as extracranial diseases or cardiogenic embolism, symptomatic ICAS (sICAS) has a higher risk of stroke recurrence, often leading to the aggravation of neurological impairment and even death. More and more evidence shows that imaging features play an important role in predicting the risk of stroke recurrence and individualized secondary prevention in patients with sICAS. This article reviews the imaging features of ischemic stroke recurrence in patients with sICAS, which provides a basis for identifying sICAS patients with high risk of recurrent stroke and effective secondary prevention.
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Abstract@#Inoculation of COVID-19 vaccines is an important approach to preventing SARS-CoV-2 infections and reducing the severe disease and mortality of COVID-19. The elderly, children and adolescents, pregnant women, lactating women, patients with chronic diseases and immunocompromised individuals are considered to be susceptible to and at a high risk of COVID-19. Early, safe and effective inoculation of COVID-19 vaccines is critical for the successful building of the population immune barrier against COVID-19. This review, based on data from clinical trials, summarizes the safety and efficacy and safety of COVID-19 vaccines among special populations, so as to provide insights into COVID-19 vaccination among special populations.
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Objective:To investigate the clinical safety and efficacy of tirofiban in the treatment of hemiplegic stroke warning syndrome.Methods:Patients with hemiplegic stroke warning syndrome admitted to Jining First People's Hospital without receiving intravenous thrombolysis from January 2018 to May 2020 were enrolled retrospectively. Some patients were given tirofiban intravenous infusion for at least 24 h in acute phase, then received oral antiplatelet therapy (tirofiban group); some only received aspirin+ clopidogrel dual antiplatelet therapy (control group). The primary endpoint was muscle strength at the paralytic side and National Institutes of Health Stroke Scale (NIHSS) score at day 7 after onset. The secondary endpoint was the modified Rankin Scale (mRS) score at 3 months after onset, and ≤2 was defined as good clinical outcome. The safety endpoint was the bleeding events during treatment. Multivariate logistic regression analysis was used to determine the independent influencing factors of clinical outcome. Results:A total of 30 patients with hemiplegic stroke warning syndrome were enrolled, including 19 (63.3%) in the tirofiban group and 11 (36.7%) in the control group. There was no significant difference in baseline clinical data between the two groups, and no drug-related bleeding complications occurred during treatment. The muscle strength at paralytic side and NIHSS score at day 7 after onset, NIHSS score at discharge and good clinical outcome rate at 3 months in the tirofiban group were significantly better than those in the control group, and the differences were statistically significant (all P<0.05). Multivariate logistic regression analysis showed that tirofiban was an independent protective factor for good outcome after adjusting the NIHSS score at the beginning of treatment (odds ratio 0.040, 95% confidence interval 0.040-0.449; P=0.009). Conclusions:Tirofiban is safe and effective in the treatment of patients with hemiplegic stroke warning syndrome in acute phase. It can effectively block the progress of the disease, improve the outcome of patients, and will not increase the risk of bleeding.
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Objective @#To investigate the awareness of human papillomavirus ( HPV ) vaccine and willingness to vaccinate daughters among parents of primary and middle school students, so as to provide the reference for the promotion of HPV vaccine in primary and middle school girls.@*Methods @#Using multi-stage stratified cluster sampling method, the parents of girls in in Grade Four to Nine from schools in Gongshu District of Hangzhou, Xiuzhou District of Jiaxing and Wuxing District of Huzhou were selected. A questionnaire survey was conducted to collect demographic information, HPV vaccine related knowledge and willingness to vaccinate daughters with HPV vaccines. The multivariate logistic regression model was used to analyze the influencing factors for the willingness to vaccinate daughters with HPV vaccines among parents. @*Results @#Totally 1 500 questionnaires were sent out, and 1 466 were effectively collected, with an effective rate of 97.73%. There were 313 fathers responded, accounting for 21.35%; and 1 153 mothers responded, accounting for 78.65%. The awareness rate of HPV vaccine was 16.81%. The rate of willing to vaccinate daughters with HPV vaccines was 49.86%. The multivariate logistic regression analysis showed that the patients who ever vaccinated daughters with self-paid vaccines ( OR=1.935, 95%CI: 1.473-2.541 ), knew cervical cancer ( OR=1.424, 95%CI: 1.065-1.904 ), knew HPV vaccine dose ( OR=1.672, 95%CI:1.216-2.301 ), knew the best vaccination period ( OR=1.392, 95%CI: 1.032-1.876 ), knew the need of cervical cancer screening even after vaccination ( OR=1.596, 95%CI:1.227-2.075) were more willing to vaccinate daughters with HPV vaccines, while the parents who thought HPV vaccine expensive ( OR=0.154, 95%CI: 0.099-0.240 ) were less willing to vaccinate daughters with HPV vaccines. @*Conclusions @#The rates of HPV vaccine awareness and willingness to vaccinate daughters are 16.81% and 49.86% among parents of primary and middle school students. Their knowledge of HPV vaccine and the price of the vaccine may affect their willingness to vaccinate daughters.
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Capsule warning syndrome (CWS) is a special clinical subtype of transient ischemic attack involving internal capsule, no cortical involvement and showing stereotyped attack. Although CWS is rare in clinical practice, most patients have poor outcomes. Therefore, a comprehensive and in-depth understanding of CWS is helpful to improve the outcomes and quality of life of patients. This article reviews the recent research progress of CWS.
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Objective@#To explore the reliability and validity of the Chinese Pediatric Quality of Life Inventory（PedsQL）in influenza children.@*Methods@#From January 2017 to February 2018,we selected laboratory-diagnosed influenza cases and healthy children according to age and gender ratio by stratified random sampling and systematic sampling method. We employed Chinese version of PedsQL 4.0 to investigate their quality of life,used Cronbach's α to evaluate the reliability,and used Pearson correlation coefficient, t-test,confirmatory factor analysis（CFA）and ROC curve to evaluate the validity. @*Results@#Totally 300 influenza cases and 300 healthy children were surveyed,with 294（98.00%）and 295（98.33%）valid questionnaires recovered. The general Cronbach's α was 0.89,and the Cronbach's α of each dimension ranged from 0.79 to 0.84. The Pearson correlation coefficients between the items and their belonged dimension ranged from 0.537 to 0.755,between the items and other dimension ranged from 0.203 to 0.384. The CFA resulted in RMSEA of 0.06,GFI of 0.88,AGFI of 0.90,CFI of 0.91 and NFI of 0.89. The scores in total and in all the dimensions in influenza cases were significantly different with those in healthy children（P<0.05）. Taking the scores in influenza cases as a golden standard,the area under the ROC curve was 0. 985（P<0.05）,the sensitivity was 0.92,and the specificity was 0.95. When the limit score was 82.18,the Youden index was the largest. @*Conclusion @#The Chinese version of PedsQL4.0 has good reliability and validity, which can be applied to quality of life assessment in children.
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@#[Abstract] Objective: To investigate the effect of alantolactone (ALT) on proliferation, migration, invasion and apoptosis of human osteosarcoma 143B cells and the underlying mechanism. Methods: Osteosarcoma 143B cells were treated with different concentrations of ALT (0, 4, 6, 8, 10 µmol/L). Then, the cell proliferation ability was detected by crystal violet staining and MTT assay, cell migration was determined by Wound-healing test, cell invasion was analyzed by Transwell assay and cell apoptosis rate was detected by Hoechst33258 staining. The mRNA and protein expressions of E-cadherin, N-cadherin, caspase-3, cleaved caspase-3 (c-caspase-3), poly ADP-ribose polymerase (PARP) and cleaved PARP (c-PARP) in 143B cells were detected by qPCR and Western blotting (WB), respectively. TCF/LEF (T cell lymphocyte factor/lymphoid enhancer factor) transcriptional activity was examined with Luciferase reporter gene assay. The mRNA and protein expressions of β-catenin as well as MMP-7 and c-Myc were detected by qPCR and WB, respectively. Results: ALT inhibited proliferation, migration and invasion of osteosarcoma143B cells and promoted apoptosis(P<0.05or P<0.01). After the treatment with ALT at 8, 10 µmol/L, the mRNA and protein expressions of E-cadherin and PARP, as well as the protein expressions of c-caspase-3 and c-PARP were up-regulated, while the mRNA and protein expressions of N-cadherin were downregulated (P<0.05 or P<0.01);At the sametime, theTCF/LEF transcriptional activity and the mRNA and protein expressions of β-catenin, MMP-7 and c-Myc were significantly down-regulated (P<0.05 or P<0.01). Conclusion:ALT may inhibit the proliferation, migration and invasion and promote cell apoptosis possibly through suppressing Wnt/β-catenin signaling pathway in osteosarcoma 143B cells.
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Intracranial atherosclerotic stenosis (ICAS) is the most common cause of ischemic stroke in Asians, and the middle cerebral artery (MCA) is its most common site. MCA stenosis can be divided into symptomatic or asymptomatic. The current endovascular treatment is mainly aimed at symptomatic stenosis. Early drug therapy can alleviate the clinical symptoms of patients and change its natural course to some extent. This article reviews the treatment of symptomatic MCA stenosis from two aspects: drug therapy and endovascular therapy.
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Posterior circulation ischemic stroke is a kind of cerebral infarction which occurs in the blood supply area of vertebrobasilar artery. Its etiology is complex. The common causes include thromboembolism, large artery atherosclerosis, and small arterial disease. The treatment options for posterior circulation ischemic stroke include drug therapy, intravenous thrombolysis and endovascular therapy. However, the application of endovascular therapy in patients with posterior circulation ischemic stroke is still in the exploratory stage, and there is not enough evidence of randomized controlled trials.
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Objective To analyze the clinical and biochemical,as well as genetic characteristics of a patient with Wiedemann-Steiner syndrome (WDSTS).Methods The clinical data of a patient with WDSTS were collected.The patient was treated with recombinant human growth hormone (rhGH) combined with gonadotrophine-releasing hormone agonist (GnRHa).Blood samples of the patient and her parents were taken for whole-Exome Sequencing (WES).Relevant literatures about KMT2A mutations were reviewed.Results The 5-year old girl presented with growth retardation,with height 100 cm (-2.4 SD),torpid reaction,and facial anomalies including low hairline,thick eyebrow and hair,hypertelorism,a wide nasal bridge.She had small and puffy hands and feet,excessive hair around back of neck,bilateral forearm and lower limbs.Her GH peak level was 26.6 ng/ml during GH stimulation test.She was re-examined at the age of 10.4 years,with severe short stature (120 cm/-3.58 SD) and a Tanner stage 2 of breast development.Her bone age was found to be approximately 11.4 years.Height increased from 120 cm at the age of 10.4 years to 147.3 cm after rhGH treatment combined with GnRHa for 2.5 years.rhGH therapy alone continued for 1.1 years and a height of 150 cm was reached at the age of 14.9 years,with bone age 14 years.Gene sequencing revealed a de novo frameshift mutation (c.10051 delA,p.Thr3351 Leufs * 17) of exon 27 in KMT2A gene of the patient,but without any mutation in her parents.Through a literature review,seventy-one patients with WDSTS (including present case) presented with intellectual disability (70/71),facial anomalies (70/71),short stature (50/71),and hypertrichosis (39/71).Conclusion Patients presented with short stature,typical facial dysmorphism,intellectual disability,and hypertrichosis should be considered for WDSTS.The mutation p.Thr3351Leufs * 17 in the KMT2A gene detected in our patient is a novel mutation.This is so far the first report of WDSTS patient who was successfully treated with a combination of GH and GnRHa at the onset of puberty to improve her adult height.
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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.
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Objectives@#To evaluate the safety of inactivated enterovirus A71(EV-A71) vaccines after large-scale immunization in the community.@*Methods@#We selected EV-A71 susceptible people (healthy children) aged 6-59 months in vaccination clinics from 89 counties in Zhejiang Province between April 2016 and March 2018. All local and systematic adverse actions were collected by 30 min on-site inspection, within 3 days and 4-30 days follow-up. Chi-square test and Fisher′s exact test were used to compare the difference of AEs incidence in various characteristics among two groups.@*Results@#A total of 71 663 doses of vaccines were included for active safety analysis, which included 37 331 doses in boys and 34 332 doses in girls. Among all the doses, children aged 6 to 11 months, 12 to 23 months and 24 to 59 months were received 13 707, 32 639 and 25 317 doses respectively. The incidence of adverse reactions within 30 min, 3 days and 4-30 days were 0.33% (239 doses), 1.58% (1 133 doses) and 0.34% (244 doses) respectively. Adverse reactions within 3 days were 1 372 doses, with a incidence of 1.91%; among all the cases, 539 doses (0.75%) were grade 1, 677 doses (0.94%) were grade 2 and 156 doses (0.22%) were grade 3, no grade-4 adverse reaction was reported. The common local adverse reactions were redness, swelling and pruritus, with the incidence rates were 0.05% (39 doses), 0.02% (16 doses) and 0.02% (12 doses), respectively, while the most common systemic adverse reaction was pyrexia with an incidence of 1.19% (856 doses), followed by diarrhea and anorexia with the incidence rates were 0.15% (104 doses) and 0.13% (90 doses) respectively.@*Conclusion@#Most adverse actions of EV-A71 vaccines were mild and moderate and majority of them were common adverse actions. No new adverse reactions were found in the study.
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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.