OBJECTIVETo learn about the pathogen spectrum and genetic characterization of HFMD with encephalitis in Yantai city.

METHODSStool samples and cerebrospinal fluids (CSF) collected from HFMD with encephalitis cases in Yantai. Virus were isolated from stool samples and identified by fluorescence reverse transcription-polymerase chain reaction. The VPl region was amplified and sequenced from positive specimens. Genetic characterization was identified by sequence analysis.

RESULTSGetting 3 virus strains from 10 stool specimens and all of them belong to EV71. The nucleotide and amino acid homogeneity with the representative isolates of C4a were 98%-99% and 98.90%-99.45% respectively.

CONCLUSIONThe pathogen of HFMD with encephalitis in Yantai city were mainly EV71 wich belong to subgenogroup C4 cluster C4a.

Capsid Proteins ; genetics ; Encephalitis, Viral ; virology ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Hand, Foot and Mouth Disease ; complications ; virology ; Humans ; Phylogeny

This study describes the expression of heat shock protein70 (HSP70) and alpha-basic-crystallin (alpha-BC) and their association with apoptosis and some related adaptor proteins in the pathogenesis of foot-and-mouth disease virus (FMDV)-induced myocarditis in lambs. HSP70 was generally overexpressed in the myocardial tissues and inflammatory cells of FMDV-induced myocarditis with differential accumulation and localization in same hearts when compared to non-foot-and-mouth disease control hearts. alpha-BC immunolabeling showed coarse aggregations in the Z line of the cardiomyocytes in FMDV-infected hearts in contrast to control hearts. Overall, the results of this study show that the anti-apoptotic proteins, HSP70 and alpha-BC, were overexpressed with increased apoptosis in FMDV-infected heart tissues. Both proteins failed to protect the cardiomyocytes from apoptosis as defense mechanisms to the FMDV during the infection, suggesting that the virus is able to increase apoptosis via both downregulation and/or upregulation of these anti-apoptotic proteins.
Animals ; Apoptosis Regulatory Proteins/metabolism ; Foot-and-Mouth Disease/*complications/*virology ; Foot-and-Mouth Disease Virus/*classification ; Gene Expression ; HSP70 Heat-Shock Proteins/*metabolism ; Myocarditis/complications/pathology/*veterinary/virology ; Myocardium/pathology ; Sheep ; Sheep Diseases/*virology ; Turkey ; alpha-Crystallin B Chain/*metabolism

Central Nervous System Diseases ; complications ; virology ; Child ; China ; epidemiology ; Enterovirus ; Enterovirus A, Human ; Female ; Hand, Foot and Mouth Disease ; complications ; epidemiology ; Humans

OBJECTIVETo identify the pathogen and characteristics on a case of hand-foot-mouth disease (HFMD) caused by coxsackie-virus A6 (CA6) associated with vaccine-derived poliovirus (VDPV) co-infection.

METHODSField epidemiological study at the epidemic area was conducted and 16 stool samples including from the patient and close contacts were collected for isolation and identification of the enterovirus (EV). 21 stool samples from patients diagnosed as HFMD were collected in the same hospital at the same month to detect CA16,EV71, CA6 and PV by real-time RT-PCR or RT-PCR. The VP1 gene of the CA6 was amplified by RT-PCR and PCR products were sequenced and analyzed.

RESULTSThe patient showed only HFMD symptoms, but no symptoms related to acute flaccid paralysis (AFP). No EVs were isolated from 16 samples collected from the patient and close contacts. And no AFP cases were found by an active search. A total of 21 samples from patients diagnosed as HFMD were collected in the same hospital at the same month and 4 were found to be EV71, 2 were CA16 and 15 (include the patient)were CA6. Only this patient was found to have had VDPV II infection. The CA6 VP1 gene was amplified from the HFMD patient and 9 other cases from the same hospital at the same month. Nucleotide sequences of the VP1 gene among the 9 strains shared 98.9%-100.0% in homology and 96.0%-100.0% in the deduced amino acid sequences. Phylogenetic analysis of the VP1 sequences categorized the 9 strains into the same branch. There were 6 nucleotides changes including U2909A between the VP1 region of the VDPV strain of the case and Sabin II. Results from phylogenetic analysis on the VP1 sequences indicated that the VDPV strain of the case was different from other VDPVs strains isolated in the world.

CONCLUSIONThis case was a HFMD which caused by CA6 co-infection with VDPV II and the VDPV was newly discovered. HFMD symptoms of the case were caused by CA6. The reason why this case did not have AFP symptoms was probably due the protective effect of IPV vaccine. No AFP cases were found by the active search for AFP cases conducted in the area, which indicated that VDPV did not cause virus circulation in this area.

Child, Preschool ; Coinfection ; Enterovirus A, Human ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; complications ; virology ; Humans ; Poliovirus Vaccines ; adverse effects

Acupuncture Therapy ; Child, Preschool ; Gait Disorders, Neurologic ; etiology ; physiopathology ; therapy ; Hand, Foot and Mouth Disease ; complications ; virology ; Humans ; Male ; Walking

OBJECTIVETo reveal the etiological agent of hand, foot and mouth disease in children in Beijing.

METHODSThroat swabs were collected from 6 infants and young children with hand, foot and mouth disease who visited the affiliated Children's Hospital from May to June 2007. Aspirated fluid from tracheal intubatton, serum and cerebral spinal fluid (CSF) were collected from a 9 years old girl (No.4243) having central neural system complication of severe hand, foot and mouth disease and admitted to the hospital from the Emergency Department. Throat swab and aspirated fluid were inoculated into the cell lines Hep-2, MDCK and Vero for virus isolation. RNAs were extracted by Trizol from 6 throat swab specimens and aspirated fluid, serum while CSF was from that severe case (No.4243). The gene fragment from 5' UTR of enterovirus was amplified from throat swabs and aspirated fluid by reverse transcription-polymerase chain reaction (RT-PCR) with the primer pairs located at the untranslated region of all enterovirus. EV71 was identified by RT-PCR with the 2 and half primer pairs located at different parts of VP1 gene of EV71. The PCR products for VP1 encoding gene of EV71 from the specimens were sequenced and sequence analysis was performed by comparing those published VP1 genes of EV71. EV71 and CA16 specific primers were used to identify the isolates by RT-PCR and the sequences were directly determined from PCR products.

RESULTSGene fragments with expected molecular weight were amplified from all 6 throat swabs and the aspirate by the primer pairs universal for the 5' UTR of enterovirus, suggesting that these patients with hand, foot and mouth disease were infected by entorovirus. Out of these 7 specimens, 2 throat swabs and the aspirate were also showing positive for the VP1 of the EV71 by different primer sets. Sequence analysis revealed that the sequences for the amplicons from 1 throat swab (No. F4211) and the aspirate shared highest homology with those published EV71, indicating that these specimens were truly positive for EV71. The sequences amplified from these specimens shared 100% and 98.9% homology to each other and were closer to the sequences of EV71 identified from Zhejiang province than those from Taiwan and strain BrCr. Gene fragments for 5' UTR of enterovirus were obtained by RT-PCR after CPE appeared in 6 out of 7 inoculations including that aspirate fluid in Vero cell, indicating that enteroviruses were isolated from these specimens. Virus isolates from one throat swab (No. F4211) and the aspirate (No. 4243) were positive by RT-PCR with the primer pairs for EV71, which was consistent with RT-PCR amplification directly from specimens. Virus isolates from other 4 specimens were CA16 by RT-PCR and sequence analysis.

CONCLUSIONThese data suggested that hand, foot and mouth disease recently appeared in children in Beijing was related with EV71 and CA16. EV71 could cause severe clinical manifestations with central nerve system complications even in the child older than 5 years.

Animals ; Cercopithecus aethiops ; Child ; China ; epidemiology ; Enterovirus A, Human ; genetics ; isolation & purification ; Enterovirus Infections ; complications ; epidemiology ; Female ; Hand, Foot and Mouth Disease ; etiology ; virology ; Humans ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, RNA ; Vero Cells ; Virus Cultivation

OBJECTIVETo evaluate the adrenocortical function in children with severe and critical enterovirus 71 infection by using a high-dose (250 µg) adrenocorticotropic hormone (ACTH) stimulation test. And to at provide experimental basis for glucocorticoid in the treatment of hand-foot-and-mouth disease (HFMD).

METHODThis was a prospective multi-center study which was carried out in PICUs of Beijing Children's Hospital, Zhengzhou Children's Hospital, Kaifeng Children's Hospital and Linyi People's Hospital in Shandong province. Children with severe and critical hand-foot-mouth disease admitted to PICUs of the four hospitals from June 2009 to April 2010 were enrolled in this study, and EV71 virus nucleic acid test and high-dose (250 µg) ACTH stimulation started at the same time. EV71 virus nucleic acid positive 51 cases were eventually enrolled in the study. Cortisol test was performed at baseline (T0) and after high-dose (250 µg) ACTH stimulation at 30 minutes (T30), 60 minutes (T60) in the first 6 hours after admission, but before glucocorticoid was given. The adrenocortical function was evaluated according to ΔTmax [ΔTmax=(T30, T60 maximum)-T0]. Diagnostic criteria of adrenal insufficiency (AI) is increment (ΔTmax)≤9 µg/dl.

RESULTThe incidence of AI in 51 cases was 52.94% (27/51). The incidence of AI in severe group was 44.74% (17/38), which was significantly higher in critical group 76.92% (10/13), P<0.05. Of the cases with a pediatric critical illness score (PCIS)≤70, 81.82% (9/11) had adrenal insufficiency, and it was 28.57% (4/14) when PCIS≥90. The incidence of AI was 75% (6/8) and 48.84% (21/43) in death and survivor group respectively, but there were no significant difference between the two groups (P>0.05). Baseline (T0) cortisol in death group was higher than survivor group (P<0.05).

CONCLUSIONAI may occur in children with enterovirus 71 infection. The critical enterovirus 71 infection had a high incidence of AI. AI may affect the prognosis of patients with severe and critical enterovirus 71 infection. Exogenous glucocorticoids administration may be considered when AI is identified or highly suspected. The timing, dosage and regimen of glucocorticoid are still unclear. Further animal experiments and clinical trials are needed.

Adrenal Insufficiency ; drug therapy ; etiology ; Adrenocorticotropic Hormone ; administration & dosage ; therapeutic use ; Child, Preschool ; Enterovirus A, Human ; pathogenicity ; Female ; Hand, Foot and Mouth Disease ; complications ; drug therapy ; physiopathology ; virology ; Humans ; Infant ; Male ; Prognosis ; Prospective Studies

In 2009, the first outbreak of hand, foot and mouth disease (HFMD) or herpangina (HP) caused by enterovirus 71 occurred in the Republic of Korea. This study inquired into risk factors associated with complications of HFMD or HP. A retrospective medical records review was conducted on HFMD or HP patients for whom etiologic viruses had been verified in 2009. One hundred sixty-eight patients were examined for this investigation. Eighty patients were without complications while 88 were accompanied by complications, and 2 had expired. Enterovirus 71 subgenotype C4a was the most prevalent in number with 67 cases (54.9%). In the univariate analysis, the disease patterns of HFMD rather than HP, fever longer than 4 days, peak body temperature over 39degrees C, vomiting, headache, neurologic signs, serum glucose over 100 mg/dL, and having an enterovirus 71 as a causative virus were significant risk factors of the complications. After multiple logistic analysis, headache (Odds ratio [OR], 10.75; P < 0.001) and neurologic signs (OR, 42.76; P < 0.001) were found to be the most significant factors. Early detection and proper management of patients with aforementioned risk factors would be necessary in order to attain a better clinical outcome.
Adolescent ; Adult ; Blood Glucose/analysis ; Body Temperature ; Enterovirus A, Human/genetics/isolation & purification ; Female ; Fever/etiology ; Genotype ; Hand, Foot and Mouth Disease/*complications/virology ; Headache/etiology ; Herpangina/*complications/virology ; Humans ; Logistic Models ; Male ; Middle Aged ; Odds Ratio ; Republic of Korea ; Retrospective Studies ; Risk Factors ; Vomiting/etiology ; Young Adult