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.

Using Interrupted Time Series Design to Analyze Changes in Hand, Foot, and Mouth Disease Incidence during the Declining Incidence Periods of 2008-2010 in China

Objective To identify patterns of hand, foot and mouth disease （HFMD） incidence in China during declining incidence periods of 2008, 2009, and 2010. Methods Reported HFMD cases over a period of 25 months were extracted from the National Disease Reporting System （NDRS） and analyzed. An interrupted time series （ITS） technique was used to detect changes in HFMD incidence rates in terms of level and slope between declining incidence periods of the three years. Results Over 3.58 million HFMD cases younger than 5 years were reported to the NDRS between May 1, 2008, and May 31, 2011. Males comprised 63.4% of the cases. ITS analyses demonstrated a significant increase in incidence rate level （P〈0.0001） when comparing the current period with the previous period. There were significant changes in declining slopes when comparing 2010 to 2009, and 2010 to 2008 （all P〈O.O05）, but not 2009 to 2008. Conclusion Incremental changes in incidence rate level during the declining incidence periods of 2009 and 2010 can potentially be attributed to a few factors. The more steeply declining slope in 2010 compared with previous years could be ascribed to the implementation of more effective interventions and preventive strategies in 2010. Further investigation is required to examine this possibility.

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.

Evaluation on Common Hand, Foot and Mouth Disease Treated by Integrative Traditional Chinese Medicine and Western Medicine

Objective To evaluate the efifcacy and safety of traditional Chinese medicine (TCM) combined with Western medicine in the treatment of patients with common hand, foot and mouth disease (HFMD) by conducting a prospective, controlled, and randomized trial. Methods A total of 452 patients with common HFMD were randomly assigned to receive Western medicine alone (n=220) or combined with TCM (Reduning or Xiyanping injections) (n=232). The primary outcome was the incidence rate of rash/herpes disappearance within 5 days, while secondary outcomes included the incidence rate for fever, cough, lethargy, agitation, and vomiting clearance within 5 days. Results The rash/herpes disappearance rate was 45.5% (100/220) in Western medicine therapy group, and 67.2% (156/232) in TCM and Western medicine combined therapy group, with significant difference (P<0.001). Moreover, TCM remarkably increased the incidence rate of secondary disappearance, which was 56.4%in Western medicine therapy group and 71.4%in TCM and Western medicine combined therapy group (P=0.001). No drug-related adverse events were observed. Conclusions It’s suggested that the integrative TCM and Western medicine therapy achieved a better therapeutic efficacy. TCM may become an important complementary therapy on relieving the symptoms of HFMD.

Investigation on Factors Associated with Severe Hand, Foot and Mouth Disease

Objective To analyze the clinical and laboratory features of patients with mild and severe HFMD to identify early predictive or diagnostic markers for severe cases. Methods Samples of feces, nasopharyngeal-swab specimens, peripheral blood, serum and cerebral spinal fluid were collected. Postmortem pathological examination was conducted on 2 dead patients with complication due to neurogenic pulmonary edema. Reverse transcription-polymerase chain-reaction(RT-PCR), culture and isolation of enterovirus 71(EV71) were performed to detect EV71 infection. Both univariate and multivariate logistic analysis were used to identify factors associated with severe cases. Results EV71 was mainly responsible for HFMD. In this study, 5 isolated EV71 strains belonged to C4 gene subtype. Compared with mild patients, EV71-RNA detection rate was higher and Cox A16 detection rate was lower among severe patients(P < 0.01). Inflammatory cell infiltration in the lung, cardiac and liver tissues were mild by postmortem pathological examination. It was found that body temperature, vomitting, limb tremor, neutrophil, blood glucose and EV71 infection were significantly related to the severe cases by univariate logistic analysis. However, after multivariate logistic regression analysis, only vomiting(OR 16.1, CI 2.3-110.5, P < 0.01) and limb tremor(OR 117.6, CI 13.8-1004.5, P < 0.01) were significantly and independently correlated with the severe cases.Conclusions EV71 was mainly responsible for HFMD, particularly for severe cases. Vomiting and limb tremor were predictive markers for severe cases.

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.

Computational Investigation of Hand Foot Mouth Disease Dynamics with Fuzziness

The first major outbreak of the severely complicated hand,foot and mouth disease(HFMD),primarily caused by enterovirus 71,was reported in Taiwan in 1998.HFMD surveillance is needed to assess the spread of HFMD.The parameters we use in mathematical models are usually classical mathematical parameters,called crisp parameters,which are taken for granted.But any biological or physical phenomenon is best explained by uncertainty.To represent a realistic situation in any mathematical model,fuzzy parameters can be very useful.Many articles have been published on how to control and prevent HFMD from the perspective of public health and statistical modeling.However,few works use fuzzy theory in building models to simulateHFMDdynamics.In this context,we examined anHFMD model with fuzzy parameters.A Non Standard Finite Difference(NSFD)scheme is developed to solve the model.The developed technique retains essential properties such as positivity and dynamic consistency.Numerical simulations are presented to support the analytical results.The convergence and consistency of the proposed method are also discussed.The proposed method converges unconditionally while the many classical methods in the literature do not possess this property.In this regard,our proposed method can be considered as a reliable tool for studying the dynamics of HFMD.

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,

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.

Value of serum pattern recognition receptors Collectin, Dectin and CD14 detection for disease evaluation in children with hand, foot and mouth disease

Objective:To study the value of serum pattern recognition receptors Collectin, Dectin and CD14 detection for disease evaluation in children with hand, foot and mouth disease (HFMD). Methods:76 children diagnosed with hand, foot and mouth disease in our hospital between May 2013 and March 2016 were selected as the HFMD group of the research, 80 healthy children receiving physical examination in our hospital during the same period were selected as the control group of the research, and the serum was collected to determine the levels of pattern recognition receptors, immunoglobulins, complements, inflammatory media and target organ damage indexes. Results:Serum Collectin, Dectin and CD14 levels of HFMD group were significantly higher than those of control group (P<0.05);serum immunoglobulin G (IgG) IgM, IgA, C3 and C4 levels of HFMD group were significantly lower than those of control group (P<0.05) and negatively correlated with Collectin, Dectin and CD14 levels;serum PCT, C-reactive protein (CRP), interleukin-6 (IL-6), IL-10, tumor necrosis factor-α(TNF-α), N-terminal pro-brain natriuretic peptide (NT-proBNP), neuron-specific enolase (NSE), S100βand surfactant protein A (SP-A) levels of HFMD group were significantly higher than those of control group (P<0.05) and positively correlated with Collectin, Dectin and CD14 levels. Conclusions:High expression of serum pattern recognition receptors Collectin, Dectin and CD14 in children with hand, foot and mouth disease is closely related to the immune response, inflammatory response and target organ function damage during the disease progression.

Hand Foot and Mouth Disease Due to Enterovirus 71 in Malaysia

Hand foot and mouth disease is a febrile sickness complex characterized by cutaneous eruption (exanthem) on the palms and soles with simultaneous occurrence of muco-cutanous vesiculo-ulcerative lesions (enanthem) affecting the mouth. The illness is caused by a number of enteroviruses with coxsackievirus A16 and enterovirus 71 as the main causative agents. Human enterovirus 71 (EV71) belongs to the species Human enterovirus A under the genus Enterovirus within the family Picornaviridae. EV71 has been associated with an array of clinical diseases including hand foot and mouth disease (HFMD), aseptic meningitis, encephalitis and poliomyelitis-like acute flaccid paralysis. A large outbreak of HFMD due to highly neurovirulent EV71 emerged in Malaysia in 1997, and caused 41 deaths amongst young children. In late 2000, a recurrence of an outbreak of HFMD occurred in Malaysia with 8 fatalities in peninsular Malaysia. Outbreak of HFMD due to EV71 recurred in 2003 with an unknown number of cases and mortalities. A similar outbreak of HFMD with 2 recorded deaths in young children occurred in peninsular Malaysia in late 2005 and this was followed by a larger outbreak in Sarawak (Malaysian Borneo) with 6 reported fatalities in the early part of 2006. The current on-going outbreak of HFMD started in peninsular Malaysia in epidemiological week 12 of 2010. As with other HFMD outbreaks in Malaysia, both EV71 and CA16 were the main aetiological viruses isolated. In similarity with the HFMD outbreak in 2005, the isolation of CA16 preceded the appearance of EV71. Based on the VP1 gene nucleotide sequences, 4 sub-genogroups of EV71 (C1, C2, B3 and B4) co-circulated and caused the outbreak of hand, foot and mouth disease in peninsular Malaysia in 1997. Two sub-genogroups (C1 and B4) were noted to cause the outbreak in 2000 in both peninsular Malaysia and Sarawak. EV71 of sub-genogroup B5 with smaller contribution from sub-genogroup C1 caused the outbreak in 2003. In the 2005 outbreak, besides the EV71 strains of sub-genogroup C1, EV71 strains belonging to sub-genogroup B5 were isolated but formed a cluster which was distinct from the EV71 strains from the sub-genogroup B5 isolated in 2003. The four EV71 strains isolated from clinical specimens of patients with hand, foot and mouth disease in the Sarawak outbreak in early 2006 also belonged to sub-genogroup B5. Phylogenetic analysis of the VP1 gene suggests that the EV71 strains causing the outbreak in Sarawak could have originated from peninsular Malaysia. Epidemiological and molecular data since 1997 show the recurrence of HFMD due to EV71 in Malaysia every 2 to 4 years. In each of the past outbreaks, more than one sub-genogroup of the virus co-circulate.

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.

Predictive correlation test of acute respiratory failure in children with hand foot mouth disease

Objective:To investigate the predictive correlation test of acute respiratory failure in children with hand foot mouth disease (HFMD).Methods: Selected from our hospital June 2014 to June 2017 HFMD incidence of acute respiratory failure in 31 cases as a observation group;Selected from our hospital same period HFMD patients without acute respiratory failure in 35 cases as the control group;the other is selected from healthy children 40 cases as normal group. There were collected three groups peripheral venous blood 2 mL, and serum was isolated. Enzyme linked immunosorbent assay was used for the determination of IL-6. The content of CRP was measured by Backman OLYMPUS AU640 automatic biochemical analyzer. The contents of creatine kinase (CK), creatine kinase isozyme (CK-MB) and lactate dehydrogenase (LDH) were determined by Backman OLYMPUS AU640 automatic biochemical analyzer. The content of troponin I (cTnI) was measured by Backman ACCESS2 chemiluminescence immunoanalyzer.Results:The serum levels of IL-6, CRP, CK, CK-MB, LDH and cTnI in the observation group were higher than those in the control group and the normal group. After treatment serum levels of IL-6, CRP, CK, CK-MB, LDH and cTnI were lower than those before treatment, and there were statistically significant differences.Conclusion: serum IL-6, CRP index and serum CK, CK-MB, LDH and cTnI indexes can be used as indicators for the prediction of acute respiratory failure in children with hand foot mouth disease, and have important clinical significance.

Association of IFNGR2 and IL-12B gene polymorphisms with susceptibility to hand,foot and mouth disease infected by EV71

Objective:To study the relationship between the single nucleotide polymorphisms(SNPs)of IFNGR2genes(rs9808753,rs11910627 and rs1532) and IL-12 B gene(rs2288831)and the susceptibility to hand,foot and mouth disease(HFMD)caused by enterovirus 71(EV71).Methods:Blood samples were collected from 145 HFMD children infected by EV71,104 children with EV71 covert infection,and 89 normal control children,followed by DNA extraction.IFNGR2 and IL-12 Bpolymorphisms were detected by imLDRTM.Results:Compared with the control group,IFNGR2rs9808753 genotype and allele distributions of EV71-infected group and EV71 covert infection group showed significant differences(P<0.05).A allele frequency in EV71-infected patients and covert EV71-infected patients was higher than that in the control group(P <0.05).While the genotype and allele frequencies of rs2288831 in IL-12 B did not show significant difference between EV71-infected groups and healthy controls.Conclusion:These findings suggested that the single nucleotide polymorphism of IFNGR2 gene site rs9808753(A/G)was associated with the susceptibility to EV71-infection,and A allele of rs9808753 might be one potential genetic susceptibility factor of EV71-infection.While the rs2288831(C/T)polymorphism of IL-12 Bdid not correlate with the susceptibility of EV71-infection.

Establishment of Indirect ELISA Diagnosis Technique based on the VP1 Protein of Foot and Mouth Disease Virus Serotype A

The VP1 protein of foot-and-mouth disease virus serotype A was prokaryotically expressed and purified to replace the traditional virus antigen for estab- lishing a fast, safe, effective indirect ELISA method, so as to detecting antibody of foot-and-mouth disease virus serotype A. Western-Blot test showed that the VP1 recombinant protein could be used as detective antigen as it can be specifically recognized by bovine positive serum of FMDV serotype A. By employing matrix titra- tion method, the optimal parameters were obtained as follows： 1 mg/L VP1 protein as coating antigen, Vserum：Vblocking solution = 1：50 dilution for serum and Vsecondary enzyme-linked antibedies：Vblocking solution ---1：2 000 for enzyme combined antibodies. The results showod that the sensitivity and specificity of this method were 94.32% and 99.09% respectively, the coefficients of variations in intra-assay and inter-assay reproducibility tests was lower than 8%. Compared with liquid phase blocking ELISA kits, the agreement of 201 serum samples reached 92.54%. The VP1-ELISA method established here is specific, sensitive, stable and simple, which can be used to monitor the antibody level of FMD serotype A.

Results on Pathogen Detection of Foot and Mouth Disease in Guangxi China and Analysis on Its Popular Spectrum

Objective: The study aims to understand the characteristics and epidemic trend of the pathogen of hand, foot and mouth disease (HFMD) in Guangxi regions, China. Besides, it aims to analyze the differences of intestinal virus detection rate between anal swab and pharyngeal swab samples. Methods: Anal swab and pharyngeal swabs of suspected HFMD children were collected in our hospital from 2012 to 2015. Real-time fluorescent PCR (Polymerase Chain Reaction) was used to detect enterovirus 71 (EV71), coxsackie virus type 16 (CA16), and universal intestinal virus nucleic acid (EV). Composition and conversion of predominant pathogens were analyzed, and paired samples’ test results of swabs anal and pharyngeal swab were statistically analyzed. Results: There are 681 cases with enterovirus in 2351 cases of patients. Among those who got enterovirus, there are 501 cases of EV71, 102 cases of CA16 and 79 cases of EV. From 2012 to 2015, the total proportion of the virus detection is 46.47%, 16.23%, 41.02% and 15.33% respectively in each year, while the proportion of predominant epidemic virus is 93.93% of EV71, 66.12% of CA16, 89.30% of EV71 and 98.73% of EV, non-EV71, non-CA16 EV (from October to December in 2015). It’s obvious that the total virus detection rate in 2012 and 2014 is significantly higher than that in 2013 and 2015. There is statistical significance. Conclusion: The main HFMD pathogens are EV71 from 2012 to 2015 in Guangxi regions. In 2012 and 2014, the predominant epidemic pathogens were EV71, while in 2013 and 2015, the predominant epidemic pathogens turn to be CA16 and non-EV71, non-CA16 EV respectively. What’s more, collecting anal swab and pharyngeal swab virus at the same time for nucleic acid detection is of great significance to improve the HFMD laboratory diagnostic.

Characterization of Hand,Foot,and Mouth Disease in China between 2008 and 2009

Objective To investigate the epidemiological and cl （HFMD） since several outbreaks of HFMD caused between 2007 and 2008. inical features of hand, foot and mouth disease by enteroviruses were documented in China Methods HFMD cases reported to the National Infectious Disease Information Management System database between May 2008 and April 2009 were assessed. Clinical features in some of the severe and fatal cases were analyzed the etiology of the outbreaks was investigated. Results 89.1% of reported HFMD cases were found in children〈5 year-old with an age-specific incidence rate of 834.1/100 000 in the first year as the notifiable disease in China from May 2008 to April 2009. The incidence, mortality and percentage of severe cases were studied for three regions of China and found to be highest in the central region. The incidence of severe cases and mortality in rural population were significantly higher than those in urban population. Among the laboratory confirmed EV17 positive cases there were 52.6% mild, 83.5% severe, and 96.1% fatal cases. More myoclonic jerks were found in the severe case group than in group that died. Tachypnea, lip purpling, pink foaming and low limb temperature occurred more frequently in the fatal cases than in the severe cases. Conclusion The epidemic of HFMD in China was characterized predominantly by EV71 infections, had relatively high mortality rates especially in the central region, and was most prevalent in young, rural populations.

Clinical characteristics of hand, foot and mouth disease in Harbin and the prediction of severe cases

Background Hand, foot and mouth disease （HFMD） is an emerging public health problem in China, not only threatening the health of children, but also causing tremendous loss and burden to both families and society. The aim of this study was to characterize the epidemiology and clinical features of HFMD, and to understand the key factors affecting HFMD in the Harbin region to provide scientific evidence for effective prevention and control strategies. Methods Epidemiological and clinical information from 2379 randomly chosen cases of HFMD treated at the Harbin Center for Disease Control and Prevention from May 2008 to November 2011 were analyzed. All cases were separated into common and severe HFMD, with key factors for severe HFMD analyzed using multivariable Logistic regression. Results Among the 2379 patients, 1798 were common cases and 581 severe cases, 14 of which resulted in death. Most cases were in children younger than 5 years. Morbidity peaked in July and was higher in the surrounding country and cities than in Harbin proper. Medical expenses were significantly higher for severe than for common cases （P 〈0.001）. The primary clinical symptoms were fever and erythema; laboratory examination showed leucocytosis together with pneumonia, carditis, and abnormal electrocardiogram and electroencephalogram in severe cases. Multivariabie Logistic regression analysis showed that the key factors for severe HFMD were age, morbidity location, morbidity area, fever duration, mouth mucosal symptoms, and abnormal serum levels of neutrophils （NEUT）, hemoglobin and glucose （P 〈0.05）. Conclusions To improve prognosis, reduce medical expense and prevent the development of severe cases, we should improve the epidemiological detection of HFMD to treat patients quickly. We should also closely monitor children with the EV71 virus, who present with continuous fever as well as abnormal laboratory results, from areas highly susceptible to

Pattern Classification of Enterovirus 71-Associated Hand, Foot, and Mouth Disease in Chinese Medicine: A Retrospective Study in 433 Cases

Objective： To determine whether patterns of enterovirus 71（EV71）-associated hand, foot, and mouth disease（HFMD） were classified based on symptoms and signs, and explore whether individual characteristics were correlated with membership in particular pattern. Methods： Symptom-based latent class analysis（LCA） was used to determine whether patterns of EV71-HFMD existed in a sample of 433 cases from a clinical data warehouse system. Logistic regression was then performed to explore whether demographic, and laboratory data were associated with pattern membership. Results： LCA demonstrated a two-subgroup solution with an optimal fit, deduced according to the Bayesian Information Criterion minima. Hot pattern（59.1% of all patients） was characterized by a very high fever and high endorsement rates for classical HFMD symptoms（i.e., rash on the extremities, blisters, and oral mucosa lesions）. Non-hot pattern（40.9% of all patients） was characterized by classical HFMD symptoms. The multiple logistic regression results suggest that white blood cell counts and aspartate transaminase were positively correlated with the hot pattern（adjust odds ratio=1.07, 95% confidence interval： 1.006–1.115; adjust odds ratio=1.051, 95% confidence interval： 1.019–1.084; respectively）. Conclusions： LCA on reported symptoms and signs in a retrospective study allowed different subgroups with meaningful clinical correlates to be defined. These findings provide evidence for targeted prevention and treatment interventions.

Construction of Plant Expression Vector for Hand,Foot and Mouth Virus EV71-VP1 Gene and Its Expression in Tomato

EV71-type virus is one of the main pathogens causing the occurrence of hand,foot and mouth disease(HFMD),and VP1 protein,a factor that directly determines the antigenicity of the virus,has been isolated.The tomato was selected as a bioreactor for the production of an edible EV71 vaccine designed for the VP1 capsid protein.Using molecular biology techniques,the fusion gene EV71-VP1 was cut from vector PGEX-4T-2,a vector containing the p2300-EV71 gene with CaMV35S promoter and TL regulatory elements was constructed,and the hypocotyl and cotyledons of tomato were transformed using Agrobacterium(EHA105)-mediated method,screened,elongated and rooted,and finally 20 resistant tomato plants were obtained.Five transgenic positive seedlings were obtained by digestion and PCR assay,among which three plants were detected by RT-PCR to be capable of transcriptional translation at the RNA level.The experimental results aimed to explore new material support for the preparation of transgenic plant oral vaccines against EV71 infection and provide a theoretical basis for accelerating the development of transgenic plant vaccines in the future.