Transmission Dynamics and Optimal Control Strategies of a Hand-Foot-Mouth Disease Model with Treatment and Vaccination Interventions

In this article, the transmission dynamics of a Hand-Foot-Mouth disease model with treatment and vaccination interventions are studied. We calculated the basic reproduction number and proved the global stability of disease-free equilibrium when R0 R0 > 1. Meanwhile, we obtained the optimal control strategies minimizing the cost of intervention and minimizing the infected person. We also give some numerical simulations to verify our theoretical results.

Expression and clinical significance of pattern recognition receptor-associated genes in hand, foot and mouth disease

Objective:To explore which pattern recognition receptors(PRRs)play a key role in the development of hand,foot,and mouth disease(HFMD)by analyzing PRR-associated genes.Methods:We conducted a comparative analysis of PRR-associated gene expression in human peripheral blood mononuclear cells(PBMCs)infected with enterovirus 71(EV-A71)which were derived from patients with HFMD of different severities and at different stages.A total of 30 PRR-associated genes were identified as significantly upregulated both over time and across different EV-A71 isolates.Subsequently,ELISA was employed to quantify the expression of the six most prominent genes among these 30 identified genes,specifically,BST2,IRF7,IFI16,TRIM21,MX1,and DDX58.Results:Compared with those at the recovery stage,the expression levels of BST2(P=0.027),IFI16(P=0.016),MX1(P=0.046)and DDX58(P=0.008)in the acute stage of infection were significantly upregulated,while no significant difference in the expression levels of IRF7(P=0.495)and TRIM21(P=0.071)was found between different stages of the disease.The expression levels of BST2,IRF7,IFI16 and MX1 were significantly higher in children infected with single pathogen than those infected with mixed pathogens,and BST2,IRF7,IFI16 and MX1 expression levels were significantly lower in coxsackie B virus(COXB)positive patients than the negative patients.Expression levels of one or more of BST2,IRF7,IFI16,TRIM21,MX1 and DDX58 genes were correlated with PCT levels,various white blood cell counts,and serum antibody levels that reflect disease course of HFMD.Aspartate aminotransferase was correlated with BST2,MX1 and DDX58 expression levels.Conclusions:PRR-associated genes likely initiate the immune response in patients at the acute stage of HFMD.

Clinical and neuroimaging features of enterovirus71 related acute flaccid paralysis in patients with hand-foot-mouth disease

Objective:To investigate clinical and neuroimaging features of enterovirus71(EV71) related acute flaccid paralysis in patients with hand-fool-mouth disease.Methods:Nine patients with acute flaccid paralysis met the criterion of EV71 induced hand-foot-mouth disease underwent spinal and brain MR imaging from May 2008 to Sep 2012.Results:One extremity flaccid was found in four cases(3 with lower limb,1 with upper limb),two limbs flaccid in three cases(2 with lower limbs,1 with upper limbs),and four limbs flaccid in two cases.Spinal MRI studies showed lesion with high signal in T2-weighted images(T2WI) and low signal T1-weighted images(T1WI) in the spinal cord of all nine cases,and the lesions were mainly in bilateral and unilateral anterior hom of cervical spinal cord and spinal cord below thoracic 9(T9) level.In addition,the midbrain,pons, and medulla,which were involved in 3 cases with brainstem encephalitis,demonstrated abnormal signal.Moreover,spinal cord contrast MRI studies showed mild enhancement in corresponding anterior hom of the involved side,and strong enhancement in its ventral root.Conclusions: EV71 related acute flaccid paralysis in patients with hand-foot-mouth disease mainly affected the anterior hom regions and ventral root of cervical spinal cord and spinal cord below T9 level. MR imaging could efficiendy show the characteristic pattern and extent of the lesions which correlated well with the clinical features.

Molecular Epidemiological Analysis of Echovirus 19 Isolated From an Outbreak Associated With Hand, Foot, and Mouth Disease (HFMD) in Shandong Province of China

To elucidate the genetic characterization and molecular epidemiological features of Echovirus 19 （El9） isolates collected from an outbreak associated with hand, foot and mouth disease （HFMD） in Tai＇an city of Shandong Province of China from July to September, 2003. Methods Thirty seven Echovirus 19 isolates were isolated from stool specimens and throat swabs collected during the outbreak, then major capsid （VP1） genomic sequence was determined, and phylogenetic tree was done based on the VP1 sequences among these 37 and other El9 viruses deposited in the Genbank. Also a representative strain named CHN-SD03-TN12 was selected for sequencing of 5′-untranslated regions （5′-UTR）. Results The identity rate was about 98.9%-100% among all these 37 El9 viruses. The genetic relationships between these 37 El9 isolates and other strains reported were also depicted. The identity rate was about 78.4%-78.9% compared with El9 reference strain Burke. The substitutions in the sequence of 5′-UTR resulted in changes in the conjectural properties of 5′-UTR of El9 viruses. Condusion The genetic features of El9 viruses isolated during the outbreak in Shandong Province in 2003 may be associated with a genetic and antigenic drift that changes the virulence of the Shandong isolates, but the molecular changes in Shandong El9 viruses contributing to their phenotype remain to be further illuminated. However, the sequences described in this paper substantiate the changes taken place in capsid VPI and 5′UTR regions. These substitutions may contribute to their tropism and virulence, and play a significant role in pathogenesis and clinical manifestations of the disease.

Application of Seasonal Auto-regressive Integrated Moving Average Model in Forecasting the Incidence of Hand-foot-mouth Disease in Wuhan,China

Outbreaks of hand-foot-mouth disease（HFMD） have occurred many times and caused serious health burden in China since 2008. Application of modern information technology to prediction and early response can be helpful for efficient HFMD prevention and control. A seasonal auto-regressive integrated moving average（ARIMA） model for time series analysis was designed in this study. Eighty-four-month（from January 2009 to December 2015） retrospective data obtained from the Chinese Information System for Disease Prevention and Control were subjected to ARIMA modeling. The coefficient of determination（R^2）, normalized Bayesian Information Criterion（BIC） and Q-test P value were used to evaluate the goodness-of-fit of constructed models. Subsequently, the best-fitted ARIMA model was applied to predict the expected incidence of HFMD from January 2016 to December 2016. The best-fitted seasonal ARIMA model was identified as（1,0,1）（0,1,1）12, with the largest coefficient of determination（R^2=0.743） and lowest normalized BIC（BIC=3.645） value. The residuals of the model also showed non-significant autocorrelations（P_（Box-Ljung（Q））=0.299）. The predictions by the optimum ARIMA model adequately captured the pattern in the data and exhibited two peaks of activity over the forecast interval, including a major peak during April to June, and again a light peak for September to November. The ARIMA model proposed in this study can forecast HFMD incidence trend effectively, which could provide useful support for future HFMD prevention and control in the study area. Besides, further observations should be added continually into the modeling data set, and parameters of the models should be adjusted accordingly.

Clinical observation on the treatment of severe hand-foot-mouth disease with antelope horn powder combined with auricular point application

Objective:To explore the Clinical observation on the treatment of severe hand-foot-mouth disease with antelope horn powder combined with auricular point application.Methods:From July 2016 to June 2018,90 children were randomly divided into control group(n=30),treatment group(n=30)and control group(n=30).The control group was treated with routine western medicine,the treatment group 1 was treated with oral antelope horn powder on the basis of control group,the second group was treated with auricular point application on the basis of treatment group 1,and the time of symptom relief and clinical cure were observed in each group.Related immune function and related inflammatory factors,serum neuron-specific enolase(NSE),safety index.Results:In treatment group 1,the time of herpes regression,antipyretic,antispasmodic time and clinical cure time were shorter than those of control group,and the time of treatment 2 group was shorter than that of group 1(P<0.05).The levels of CD3,CD4 and CD8 were increased after treatment in the three groups,especially in the treatment group(P<0.05),and the levels of TNF-毩αand IL-2,IL-6,IL-10 were decreased after treatment in the three groups,especially in the treatment group(P<0.05).NSE decreased after treatment in three groups,especially in treatment group 2(P<0.05).Conclusion:The application of antelope horn powder combined with auricular point application can obviously improve the severe hand,foot and mouth disease,and the clinical curative effect is definite.It is worth popularizing and applying in clinical practice.

Effect of interferon + Potassium Sodium Dehydroandrograpolide Succinate therapy on inflammatory response and immune response in children with hand-foot-mouth disease

Objective: To investigate the effect of interferon + Potassium Sodium Dehydroandrograpolide Succinate therapy on inflammatory response and immune response in children with hand-foot-mouth disease. Methods: A total of 116 children with hand-foot-mouth disease who were treated in this hospital between September 2016 and February 2018 were selected as the study subjects and divided into the control group (n=58) and the Potassium Sodium Dehydroandrograpolide Succinate group (n=58) by random number table method. Control group received symptomatic + interferon therapy, and Potassium Sodium Dehydroandrograpolide Succinate group received symptomatic + interferon + Potassium Sodium Dehydroandrograpolide Succinate therapy, and they were treated for 1 week. The differences in the serum contents of inflammatory factors, adhesion molecules and humoral immunity indexes were compared between the two groups before and after treatment. Results:Before treatment, serum levels of inflammatory factors, adhesion molecules and humoral immunity indexes were not significantly different between the two groups. After 1 week of treatment, serum inflammatory factors IL-1β, IL-10, PCT and hs-CRP levels of Potassium Sodium Dehydroandrograpolide Succinate group were lower than those of control group;serum adhesion molecules CD44, ICAM-1 and VCAM-1 levels were lower than those of control group;serum humoral immunity indexes IgA, IgG, C3 and C4 levels were lower than those of control group. Conclusion: Interferon + Potassium Sodium Dehydroandrograpolide Succinate therapy can effectively reduce the systemic inflammatory response and improve the humoral immune function in children with hand-foot-mouth disease.

Correlation of serum NSE and S100β levels with inflammatory response and immune response in children with hand-foot-mouth disease complicated by encephalitis

Objective:To study the correlation of serum NSE and S100β levels with inflammatory response and immune response in children with hand-foot-mouth disease (HFMD) complicated by encephalitis.Methods:Children who were diagnosed with hand-foot-mouth disease in Yulin Third Hospital between May 2015 and February 2017 were selected, children who were combined with central nervous system were selected as severe group, and children who were not combined with central nervous system were selected as mild group;children who received physical examination during the same period were selected as the control group. Serum was collected to determine the contents of NSE, S100β and inflammatory response mediators, and peripheral blood was collected to determine the contents of T cell subsets and NK cells.Results: Serum NSE and S100β levels of severe group and mild group were significantly higher than those of control group, and serum NSE and S100β levels of severe group were significantly higher than those of mild group;serum TNF-α, IL-6, IL-10, IL-17 and PCT levels as well as peripheral blood HLA-DR+CD4+, HLA-DR+CD8+, CD38+CD4+ and CD38+CD8+ levels of severe group and mild group were significantly higher than those of control group while peripheral blood CD3+T cell, CD4+T cell, CD8+T cell and NK cell levels were significantly lower than those of control group;serum TNF-α, IL-6, IL-10, IL-17 and PCT levels as well as peripheral blood HLA-DR+CD4+, HLA-DR+CD8+, CD38+CD4+ and CD38+CD8+ levels of severe group were significantly higher than those of mild group and positively correlated with NSE and S100β levels while peripheral blood CD3+T cell, CD4+T cell, CD8+T cell and NK cell levels were significantly lower than those of mild group and negatively correlated with NSE and S100β levels.Conclusion: The increase of serum NSE and S100β levels in children with HFMD complicated by encephalitis is closely related to inflammatory response activation and immune response disorder.

Molecular Epidemiology of Enteroviruses Associated with Hand, Foot, and Mouth Disease in the Mainland of China

Hand, foot, and mouth disease （HFMD） is a common contagious illness which occurs worldwide both sporadically and in epidemics. The disease mainly affects, children and the typical symptoms, which may resolve spontaneously, include mucocutaneous papulovesicular lesions on the hands, feet, mouth, and buttocks. In rare cases, however, the patients may also develop neurological complications such as neurogenic pulmonary edema,

A Novel Early Warning Model for Hand, Foot and Mouth Disease Prediction Based on a Graph Convolutional Network

Objectives Hand,foot and mouth disease(HFMD)is a widespread infectious disease that causes a significant disease burden on society.To achieve early intervention and to prevent outbreaks of disease,we propose a novel warning model that can accurately predict the incidence of HFMD.Methods We propose a spatial-temporal graph convolutional network(STGCN)that combines spatial factors for surrounding cities with historical incidence over a certain time period to predict the future occurrence of HFMD in Guangdong and Shandong between 2011 and 2019.The 2011-2018 data served as the training and verification set,while data from 2019 served as the prediction set.Six important parameters were selected and verified in this model and the deviation was displayed by the root mean square error and the mean absolute error.Results As the first application using a STGCN for disease forecasting,we succeeded in accurately predicting the incidence of HFMD over a 12-week period at the prefecture level,especially for cities of significant concern.Conclusions This model provides a novel approach for infectious disease prediction and may help health administrative departments implement effective control measures up to 3 months in advance,which may significantly reduce the morbidity associated with HFMD in the future.

Chinese herbal medicine for hand-foot-and-mouth disease in children: An overview of systematic reviews

Objective:To summarize the use of Chinese herbal medicines (CHM) for the treatment of hand-foot-and-mouth disease (HFMD) in children and to provide high-level evidence for clinical decision-making.Methods:We conducted an overview of systematic reviews (SRs).Two English-language and four Chinese-language electronic databases were searched from inception to March 31,2018.Published SRs and meta-analyses evaluating CHM use in children with HFMD and reporting clinically-relevant outcomes such as time to fever resolution were eligible for inclusion in this overview.Reviews were accepted if the intervention featured CHM with or without other treatment.Two authors evaluated the methodological quality of the included SRs by using ASMTAR and ROBIS.Results:Thirty SRs comprising 90 244 children with HFMD were included in this overview of SRs.All SRs were published in Chinese between 2011 and 2017.All intervention arms received CHM with or without conventional treatment and were compared against control arms receiving no treatment,conventional treatment alone,or placebo.All 30 reviews reported the time to fever resolution and results demonstrated that CHM had a greater benefit in reducing fever compared with controls.Twenty-three reviews reported the time to rash resolution which presented similar results.Fourteen SRs reported the time to oral ulcer healing and CHM demonstrated a significantly reduced time to resolution compared to controls.The quality of the included SRs was low to moderate as assessed by the AMSTAR tool.Conclusion:Published reviews demonstrated potential benefits of CHM in children with HFMD.Overall,the methodological quality of reviews included in this overview of SRs was low and our findings should be interpreted with caution.We would strongly recommend that future SRs be designed and reported rigorously following PRISMA in order to provide more robust evidence on which to base clinical guidance.

The Comparative Study on Two Models of Syndrome Differentiation of the Hand, Foot and Mouth Disease: An Investigation Analysis of the Signs and Symptoms on 2325 Cases

Objective To realize the characteristics of "zheng" differentiation-treatment for hand, foot and mouth disease(HFMD), a new methodology of syndrome differentiation for different stages of HFMD has been explored. Methods Total of 2 325 cases with HFMD were recorded by distributing them into exterior syndrome stage, interior syndrome stage, severe syndrome stage and recovered syndrome stage, respectively, and the main symptoms and subsidiary symptoms of different stages of HFMD have been observed. The major and minor pathogenesis of HFMD in different stages were obtained, and compared with the "2010 Guideline for the Diagnosis and Treatment of HFMD". Results It was found that the major pathogenesis of exterior stage was defined as "the invation of the wenevil to the defender of the body with the collaterals got involved ", and the minor as "qi deficiency"; in interior stage, "the fury of Gan-Yang" was the main pathogenesis, and "qi in chaos and qi deficiency" was the minor; in severe syndrome stage, "the damage of heart, liver and lung" was the main pathogenesis, and "qi in chaos" was the minor; and the pathogenesis of recovered stage was "qi-yin deficiency". Compared with the "2010 Guideline for the Diagnosis and Treatment of HFMD", it showed that "the obstruction of the fei-pi qi by the mixture of shi-re evil" and "the mixture of shi-re" in vivo was quite difficult to be explained in completely different context in the general situation; in the severe stage, the TCM clinical characteristics of syndrome differentiation might lose; in the early acute severe cases, the phenomenon that xin-yang and fei-qi almost ran out was difficult to be observed, then, the line between the severe and the acute severe became vague.Conclusions The theory of syndrome differentiation by stages of HFMD was reasonable in the actual situation of clinical description on HFMD which was expected to be further tested and widely applied in the "zheng" differentiation-treatment of HFMD in the future.

Prevalence analysis of hand,foot and mouth disease in China,2008-2018

Objective:To analyze the prevalence of hand,foot and mouth disease(HFMD)in China during 2008-2018 and to provide reference for the prevention and control of HFMD.Methods:Literature eligible for selection was retrieved from the China's national knowledge infrastructure project(CNKI),and data from 2008-2018 Chinese HFMD reports were systematically analyzed.Excel and SPSS 20.00 were used for statistical analysis,and GraphPad Prism 6 was used for mapping.Results:The analysis showed that the average incidence rate of HFMD in China from 2008 to 2018 was 123.37±93.14 cases per 100,000 people,with a male-to-female ratio of(1.50±0.15):1,and the vast majority of cases were concentrated in the under-5 age group,mainly in scattered children.The time of onset of HFMD showed obvious seasonal characteristics,in which single-peak provinces were higher than double-peak provinces,with double-peak trends in April-July and October-November,respectively,and single-peak trends mainly in May-July.Based on the spatial distribution study,the incidence pattern showed obvious regional differences(P<0.05),with high incidence concentrated in the southeastern part of the country,and a gradual increase in incidence from west to east and north to south of China.EV71,CA16 and other enteric viruses were the main pathogens of HFMD,with EV71 accounting for 53.5%,CA16 is accounting for 14.3%and other enteric viruses accounting for 32.2%of the pathogens.Conclusions:The incidence of HFMD in China is high,with obvious geographical,seasonal,gender and age distribution characteristics.According to the pattern of HFMD incidence in the region,early prevention and control of key areas and groups is an effective way to cut off the spread of the disease.

Regional characteristics and spatiotemporal differentiation of the prevalence of hand, foot, and mouth disease in Xinjiang, China

In recent years,the number of hand,foot,and mouth disease(HFMD)cases in China has continued to grow,and the disease has become a serious public health issue.Xinjiang Uygur Autonomous Region is located in the arid areas of Northwest China,where the epidemiological trend of HFMD is gradually increasing and characterized by geographical heterogeneity.In this study,based on the HFMD case data in all counties and cities in Xinjiang,we employed statistical and GIS spatial analyses,and geographic probe models to characterize the spatiotemporal differentiation of HFMD epidemics in Xinjiang during the period of 2009–2018,and quantitatively analyzed the factors influencing the spatial differentiation of HFMD epidemics.The results showed that HFMD incidence rate in Xinjiang had non-stationary temporal characteristics on the interannual and monthly scales,and the monthly variation characteristics of HFMD epidemic were quite different in southern and northern Xinjiang.The spatial distribution characteristics of HFMD epidemics showed a north–south spatial differentiation pattern with the Tianshan Mountains as the boundary;cold spot and hot spot of HFMD epidemics in Xinjiang have shifted from scattered to concentrated,and the spatial differentiation pattern had gradually stabilized.Moreover,the dominant factors influencing the spatial differentiation of HFMD epidemics in Xinjiang were socioeconomic factors,such as per capita GDP and urbanization rate,while the basic factors affecting its spatial differentiation were natural environmental factors.The spatial differentiation and evolution patterns of HFMD epidemics differed between northern and southern Xinjiang.Specifically,the leading role of socioeconomic factors is more obvious in southern Xinjiang than in northern Xinjiang,while natural environmental factors(e.g.,dryness and relative humidity)contribute to the prevalence of HFMD epidemics in northern Xinjiang,and the perturbing effect of these factors was more prominent than other factors.The results of this study can provide a scientific basis for the prevention of HFMD epidemics and early warning of HFMD epidemics in Xinjiang.

天津市手足口病防控知识科普宣讲活动及效果评价

2023年6—10月,对天津市16个区进行分层抽样,抽取4个宣传地区及其24所托幼机构,对849位幼儿家长、705名校(园)医、129名社区医生开展手足口病防控知识科普宣讲活动,宣传形式包括家长课堂宣讲,校医、社区医生现场活动和答疑解惑,以科普文章制作转发、视频播放等形式传播,并针对现场宣讲活动人群开展问卷调查。科普宣讲活动后,手足口病日常预防知识知晓率及防控行为率明显升高。提出对于需要多次强化或不易记忆的日常防护知识,可采用科普宣讲和短视频循环播放相结合的方式。

深圳市宝安区手足口病经济负担及影响因素

目的:了解深圳市宝安区2021年手足口病经济负担并分析其影响因素。方法:收集某三级甲等医院2021年9月1日—12月1日就诊的1 225例手足口病病人,设计调查问卷,采用电话询问及医院信息系统查询收集资料,评估手足口病造成的经济负担并分析其影响因素。结果:1 225例病人中,门诊病人1 195例(97.55%),住院病人30例(2.45%),无重症病例。1 195例手足口病门诊病人的总体经济负担为422.57(336.70,604.54)元/例,30例手足口病住院病人的总体经济负担为(4 386.88±1 624.61)元/例。年龄、报销类型以及诊疗次数是手足口病门诊病人总体经济负担的影响因素(P<0.05)。结论:手足口病住院病人经济负担高于门诊病人,年龄、报销类型以及诊疗次数是手足口病门诊病人经济负担的影响因素。早发现、早诊断、早治疗能有效降低疾病经济负担。

上海市长宁区儿童家长肠道病毒71型灭活疫苗接种意愿及影响因素分析

目的 了解上海市长宁区托幼机构儿童家长为孩子接种肠道病毒71型灭活疫苗(EV71疫苗)的意愿并分析影响因素,为推广EV71疫苗接种提出建议。方法 采用随机抽样的方式抽取长宁区托幼机构就读的儿童,向儿童家长发放问卷进行调查。结果 有75.40%的家长愿意为孩子接种EV71疫苗。Logistic回归分析,不认为EV71会产生强烈副作用、疫苗犹豫量表得分越高、主动获取知识得分越低的家长更愿意为孩子接种EV71疫苗。结论 家长为儿童接种EV71疫苗的意愿受到对EV71疫苗副作用的认知、疫苗犹豫和主动获取知识情况的影响,应对家长开展相关健康教育,以提升接种率。

2016至2022年杭州地区14259例儿童肠道病毒流行特征分析

目的了解杭州地区手足口病(hand-foot-mouth disease,HFMD)病原体肠道病毒(entero virus,EV)的阳性检出情况及流行特征,为今后制定杭州地区HFMD防控策略提供科学依据。方法收集整理2016年1月至2022年12月期间杭州市儿童医院检出的EV通用型核酸阳性的14259例的病例信息,对年龄、性别、检出时间等因素进行回顾性分析。结果EV历年平均阳性检出率为45.48%(14259/31354),其中男童阳性检出率为27.20%,女童为18.28%,差异有统计学意义(P<0.01)。不同年龄段患儿EV阳性检出率随年龄增大呈下降趋势,5岁以下儿童的EV阳性检出率占所有阳性检出患儿的90.38%。在季节方面,EV夏季高发,历年夏季5月至7月的总阳性检出数占总检出数的49.16%,自2016年以来EV阳性检出率呈逐年下降趋势。结论杭州地区EV感染具有性别差异,男童易感染,尤其需要重视5岁以下儿童的EV防护。另外HFMD存在一定的季节性,夏季高发,历年检出率自2016年后逐年下降,可能与疫苗推广和健康意识提高存在一定关系。

Transmission dynamics of Hand-Foot-Mouth Disease with partial immunity through non-integer derivative

In this paper,we formulate the transmission phenomena of Hand-Foot-Mouth Disease(HFMD)through non-integer derivative.We interrogate the biological meaningful results of the recommended system of HFMD.The basic reproduction number is determined through next generation method and the impact of different parameters on the reproduction number is examined with the help of partial rank correlation coeficient(PRCC)technique.In addition,we concentrated on qualitative analysis and dynamical behavior of HFMD dynamics.Banach's and Schaefer's fixed-point theorems are used to analyze the uniqueness and existence of the solution of the proposed HFMD model.The HFMD system's Ulam-Hyers stability has been confirmed to be sufficient.To highlight the impact of the parameters on the dynamics of HFMD,we performed several simulations through numerical scheme to conceptualize the transmission route of the infection.To be more specific,numerical simulations are used to visualize the effect of input parameters on the systems dynamics.We have shown the key input parameters of the system for the control of infection in the society.

The Application Research of WBC Morphology Combined with NAP in the HFMD Co-Infection of Children

In this manuscript the authors have studied the application value of White blood cell morphology combined with neutrophils alkaline phosphatase staining integral value in treatment of pediatric HFMD combined with infection. They showed that comparing the NAP positive rate of HFMD co-infection with the NAP positive rate of other three groups respectively—reference group, HFMD group and anti-infection HFMD group—the t value is 25.7964, 28.2326, 28.3696 and the values of P are 0.0000;Comparing the NAP integral value of HFMD co-infection with the NAP integral value of other three groups respectively, which are reference group, HFMD group and anti-infection HFMD, the t value is 35.8687, 36.2664, 36.1374 and the values of P are 0.0000. They concluded that it is useful to apply WBC morphological examination combined with the positive rate and integral value of NAP staining to diagnose and treatment HFMD.