Molecular epidemiology of an outbreak of hand, foot, and mouth disease associated with subgenotype C4a of enterovirus A71 in Nanchang, China in 2014.

An outbreak of hand, foot, and mouth disease was reported through hospital-based surveillance in Nanchang, China in 2014. A total of 244 cases were reported, 176 (72.1%) cases were tested positive for enteroviruses by direct reverse transcription-polymerase chain reaction, in which enterovirus A71 (EV-A71), coxsackievirus A16 (CV-A16), and untyped enteroviruses (UEV) accounted for 84.1%, 3.4%, and 12.5%, respectively. In this outbreak, children under 5 years old constituted more than 98% of the positive cases, and the ratio of male to female cases was 2.6 to 1 (P < 0.01). Phylogenetic analysis indicated that the Nanchang EV-A71 strains belonged to subgenotype C4a undergoing continuously evolutionary changes.

Knowledge, Attitudes, and Practices (KAP) Relating to Avian Influenza (H10N8) among Farmers' Markets Workers in Nanchang, China.

Three cases of avian influenza virus H10N8 were reported in Nanchang, China, as of April 2014. To identify the knowledge, attitudes, and practices (KAP) related to H10N8 among farmers' market workers, a cross-sectional survey was conducted in 63 farmers' markets in Nanchang. Using the resulting data, characteristics of poultry and non-poultry workers' knowledge, attitudes, and practice were described. Results suggest that interventions targeting high-risk workers should be developed and implemented by public health agencies to prevent the spread of H10N8. Additionally policies that encourage farmers' market workers to receive influenza vaccine should be developed, adopted, and enforced.

Coexistence of Avian Influenza Virus H10 and H9 Subtypes among Chickens in Live Poultry Markets during an Outbreak of Infection with a Novel H10N8 Virus in Humans in Nanchang, China.

Infection with the novel H10N8 virus in humans has raised concerns about its pandemic potential worldwide. We report the results of a cross-sectional study of avian influenza viruses (AIVs) in live poultry markets (LPMs) in Nanchang, China, after the first human case of H10N8 virus infection was reported in the city. A total of 201 specimens tested positive for AIVs among 618 samples collected from 24 LPMs in Nanchang from December 2013 to January 2014. We found that the LPMs were heavily contaminated by AIVs, with H9, H10, and H5 being the predominant subtypes and more than half of the LPMs providing samples that were positive for the H10 subtype. Moreover, the coexistence of different subtypes was common in LPMs. Of the 201 positive samples, 20.9% (42/201) had mixed infections with AIVs of different HA subtypes. Of the 42 mixed infections, 50% (21/42) showed the coexistence of the H9 and H10 subtypes, with or without H5, and were from chicken samples. This indicated that the H10N8 virus probably originated from segment reassortment of the H9 and H10 subtypes.

Genetic diversity of avian influenza A (H10N8) virus in live poultry markets and its association with human infections in China.

Following the first human infection with the influenza A (H10N8) virus in Nanchang, China in December 2013, we identified two additional patients on January 19 and February 9, 2014. The epidemiologic, clinical, and virological data from the patients and the environmental specimen collected from 23 local live poultry markets (LPMs) were analyzed. The three H10N8 cases had a history of poultry exposure and presented with high fever (>38°C), rapidly progressive pneumonia and lymphopenia. Substantial high levels of cytokines and chemokines were observed. The sequences from an isolate (A/Environment/Jiangxi/03489/2013 [H10N8]) in an epidemiologically linked LPM showed highly identity with human H10N8 virus, evidencing LPM as the source of human infection. The HA and NA of human and environmental H10N8 isolates showed high identity (99.1-99.9%) while six genotypes with internal genes derived from H9N2, H7N3 and H7N9 subtype viruses were detected in environmental H10N8 isolates. The genotype of the virus causing human infection, Jiangxi/346, possessed a whole internal gene set of the A/Environment/Jiangxi/10618/2014(H9N2)-like virus. Thus, our findings support the notion that LPMs can act as both a gene pool for the generation of novel reassortants and a source for human infection, and intensive surveillance and management should therefore be conducted.

Investigation of avian influenza virus in poultry and wild birds due to novel avian-origin influenza A(H10N8) in Nanchang City, China.

Multiple reassortment events within poultry and wild birds had resulted in the establishment of another novel avian influenza A(H10N8) virus, and finally resulted in human death in Nanchang, China. However, there was a paucity of information on the prevalence of avian influenza virus in poultry and wild birds in Nanchang area. We investigated avian influenza virus in poultry and wild birds from live poultry markets, poultry countyards, delivery vehicles, and wild-bird habitats in Nanchang. We analyzed 1036 samples from wild birds and domestic poultry collected from December 2013 to February 2014. Original biological samples were tested for the presence of avian influenza virus using specific primer and probe sets of H5, H7, H9, H10 and N8 subtypes by real-time RT-PCR. In our analysis, the majority (97.98%) of positive samples were from live poultry markets. Among the poultry samples from chickens and ducks, AIV prevalence was 26.05 and 30.81%, respectively. Mixed infection of different HA subtypes was very common. Additionally, H10 subtypes coexistence with N8 was the most prevalent agent during the emergence of H10N8. This event illustrated a long-term surveillance was so helpful for pandemic preparedness and response.

Clinical and epidemiological characteristics of a fatal case of avian influenza A H10N8 virus infection: a descriptive study.

BACKGROUND: Human infections with different avian influenza viruses--eg, H5N1, H9N2, and H7N9--have raised concerns about pandemic potential worldwide. We report the first human infection with a novel reassortant avian influenza A H10N8 virus. METHODS: We obtained and analysed clinical, epidemiological, and virological data from a patient from Nanchang City, China. Tracheal aspirate specimens were tested for influenza virus and other possible pathogens by RT-PCR, viral culture, and sequence analyses. A maximum likelihood phylogenetic tree was constructed. FINDINGS: A woman aged 73 years presented with fever and was admitted to hospital on Nov 30, 2013. She developed multiple organ failure and died 9 days after illness onset. A novel reassortant avian influenza A H10N8 virus was isolated from the tracheal aspirate specimen obtained from the patient 7 days after onset of illness. Sequence analyses revealed that all the genes of the virus were of avian origin, with six internal genes from avian influenza A H9N2 viruses. The aminoacid motif GlnSerGly at residues 226-228 of the haemagglutinin protein indicated avian-like receptor binding preference. A mixture of glutamic acid and lysine at residue 627 in PB2 protein--which is associated with mammalian adaptation--was detected in the original tracheal aspirate samples. The virus was sensitive to neuraminidase inhibitors. Sputum and blood cultures and deep sequencing analysis indicated no co-infection with bacteria or fungi. Epidemiological investigation established that the patient had visited a live poultry market 4 days before illness onset. INTERPRETATION: The novel reassortant H10N8 virus obtained is distinct from previously reported H10N8 viruses. The virus caused human infection and could have been associated with the death of a patient. FUNDING: Emergency Research Project on human infection with avian influenza H7N9 virus, the National Basic Research Program of China, and the National Mega-projects for Infectious Diseases.

[Retrospective analysis of human avian influenza A (H10N8) virus infection in Jiangxi province].

OBJECTIVE: To analyze and discuss the source and epidemic disposition of human infection with avian influenza A (H10N8) virus. METHODS: Epidemiological surveys were used to collect related data and RT-PCR was applied to detect the specimens collected from cases, close contacts related exposure to live poultry markets. Data were analyzed descriptively. RESULTS: Three cases were discovered by surveillance on patients with severe pneumonia, two of the three died, but one in the hospital with the course over 6 months. All the three cases had histories of exposure to live poultry or related markets. Lower respiratory tract gargle aspirate samples of 3 patients were detected by Nanchang Municipal Center for Disease Control and Prevention (CDC) and Jiangxi Provincial CDC, and the results showed the influenza 2009pdmH1, H3, H5, H7, H9 subtypes negative. Specimen of patient 1 and 2 was positive for influenza A universal primers. Specimen of patient 3 was positive for H10N8 detected by Chinese National Influenza Center. All 33 close contacts of the patients were negative for H10N8 virus. Positive rate of the total poultry environmental specimens collected from the cases exposure markets was 5.19%. No new cases were found, after the prevention and control messages were implemented. CONCLUSION: Three cases of H10N8 avian influenza virus infection province might be associated with exposure to live poultry market in Jiangxi.

Molecular epidemiology of human enterovirus 71 strains in the Nanchang region of China in 2011.

Hand, foot, and mouth disease (HFMD) is caused by enteroviruses, most commonly enterovirus 71 (EV71) and coxsackievirus A16 (CA16). In general, EV71 infection is more likely to induce severe complications and mortality than other enterovirus infections. The present study focuses on the molecular epidemiology of human EV71 strains in the Nanchang region of China in 2011. Overall, 651 specimens (throat or rectal swabs) were collected, and one-step reverse transcriptase-polymerase chain reaction was performed for analysis. Enteroviruses were detected in 62.2% (405/651) of the specimens. EV71, CA16, and other enteroviruses were found in 292, 73, and 40 specimens, respectively. Phylogenetic analysis of the VP1 region of the 8 EV71 strains found in the Nanchang region indicated that these strains belong to the C4 subgenotype. This study shows that the C4 subgenotype strain of EV71 was prevalent in the HFMD cases of Nanchang in 2011, and it reports the first incidence of adults being infected by EV71 in the Nanchang region. Thus, the surveillance of HFMD epidemiology and monitoring of HFMD severity should be continued.

Nucleotide sequence characterization and phylogenetic analysis of hantaviruses isolated in Shandong Province, China.

BACKGROUND: China is the most severe endemic area of hemorrhagic fever with renal syndrome (HFRS) in the world with 30,000-50,000 cases reported annually, which accounts for more than 90% of total number of cases worldwide. The incidence rate of the syndrome in Shandong Province is one of the highest in China, which has ever reached 50 per 100,000 persons per year. However, the molecular characteristics of hantaviruses (HV) epidemic in Shandong Province remain unclear. Therefore it is useful to clarify nucleotide sequence and phylogenetic characteristics of HV isolated in Shandong Province in order to provide better advices to control and prevent HFRS. METHODS: RNAs were extracted from sera of clinically diagnosed patients and positive rodent lungs that were detected by indirect immunofluorescent assay (IFA). Partial M segments of HV were amplified from the RNAs with reverse transcription nested polymerase chain reactions (nested PCR) using hantavirus genotype specific primers. The nested PCR products were sequenced and compared with those from previously epidemic isolates in Shandong and with other representative HV sequences from GenBank. Phylogenetic tree analyses were performed based on the sequences of the M genes. RESULTS: Thirty-four HV isolates in Shandong showed 67.1%-100% nucleotide identities. The nucleotide homologies among 6 Hantaan viruses (HTNV) isolates in Shandong were 78.1%-98.7%, while the homologies among 28 Seoul virus (SEOV) isolates in Shandong were 93.7%-100%. There were at least 3 subtypes HTNV (H2, H5, H9) and 2 subtypes SEOV (S2, S3) in Shandong Province. CONCLUSIONS: In Shandong Province, the homologies of HTNV were lower and there were no predominant subtypes, while the homologies of SEOV were higher and S3 was the predominant subtype. The homologies of SEOV from rodents were higher than those from patients. The distribution of subtypes in Shandong was similar to that of the adjoining provinces. Phylogenetic analyses of the sequences showed geographic clustering of HV in Shandong.

Joint effects of eNOS gene T-786C and ADH2 Arg47His polymorphisms on the risk of premature coronary artery disease.

INTRODUCTION: Both T-786C mutation in endothelial nitric oxide synthase (eNOS) gene and alcohol dehydrogenase (ADH) gene polymorphism such as ADH3 gamma1/gamma2 have been reportedly associated with coronary artery disease (CAD). Since ADH2 Arg47His polymorphism is common in Asian population, the aim of this present study was to assess the interaction between eNOS gene T-786C and ADH2 Arg47His polymorphisms on premature CAD risk. MATERIALS AND METHODS: Hospital-based case-control study was conducted with 167 premature CAD and 235 late-onset CAD patients. Polymerase chain reaction restriction fragment length polymorphism was used to detect the polymorphisms. Multivariate logistic regression model was performed to adjust the potential confounders and estimate odds ratios (ORs) with 95% confidence intervals (CIs). Synergy index (S) was the measure to assess the interaction as departure from additivity. RESULTS: After the adjustment for the potential confounders, and compared with the carriers of TT and Arg/Arg as the reference, the ORs with 95% CIs in parentheses of premature CAD were that 1.13 (0.19-6.59) for CT or CC and Arg/Arg carriers; 2.24 (0.77-6.49) for TT and Arg/His or His/His carriers; 4.18 (1.32-13.22) for CT or CC and Arg/His or His/His carriers, respectively. Based on those ORs, S was 2.32 (95% CI: 0.37-14.72). CONCLUSIONS: The mutant genotypes of eNOS gene T-786C mutation and the fast form of ADH2 Arg47His polymorphism had an additive interaction on the risk of premature CAD in Chinese population. Further investigations with big sample size are necessary for confirming this additive interaction.

[The relationship between gene polymorphism of catechol-O-methyltransferase and survival of oral pesticides suicide attempters].

OBJECTIVE: To understand the association between gene polymorphism of catechol-O-methyltransferase (COMT) and suicide attempters taking pesticides orally and to provide a scientific base for the prevention of suicidal behavior. METHODS: 107 suicide attempters taking pesticides orally were collected from The People's Hospitals in Shandong Province from January 2003 through December 2003. The informations of all the attempters were obtained and 1 ml whole blood from of them were collected. Molecular biological techniques were used to study the gene type of each subject. RESULTS: In the 107 suicide attempters, the ratio of male to female is 1:1.55; 67.3% of them were in age range of 20 - 44 years; 59.8% of them had education level of elementary school or lower. 13 suicide attempters (12.1% of all the attempters) had psychiatric disorders. 84 suicide attempters (78.5% of all the attempters) had affective conflicts with others in the recent year. 30 suicide attempters (28.0% of the suicide attempters) had impulsive behavior. Multivariate logistic regression model analysis showed that suicide impulsion and psychiatric disorders were significantly associated with COMT Val/Val, their OR values were 0.052 (95% CI: 0.006 - 0.437), 2.917 (95% CI: 1.097 - 7.760). CONCLUSION: In this population, more attention should be paid to young female people with psychiatric disorders or having affective conflicts with others in the recent year so as to prevent suicide. This study supported that there is heterogeneity in COMT gene in suicide attempters and there is interaction between COMT Val/Val and suicide impulsion and psychiatric disorders.

[A paired case-control study on related factors to attempted suicide].

OBJECTIVE: To understand the environmental risk factors on attempted suicide, and to study the interaction between factors as gene polymorphism of catechol-O-methyltransferase (COMT) associated to attempted suicide. METHODS: Paired case-control study of 205 suicide attempters (89 male, 116 female) and molecular biological techniques were used to study the relation between gene polymorphism of COMT, environmental factors and the rate of attempted suicide. Controls were paired with cases according to the same gender, similar age (no more than 3 years) and in the same district. RESULTS: There were no significant differences in gene types and gene frequency between case and control groups. Multivariate conditional logistic regression model analysis showed that COMT Val/Val 158/108, low education level, cigarette smoking, emotional conflicts, psychologic disorders and depression were risk factors of attempted suicide with OR values as 2.43 (95% CI: 1.10 - 5.40), 5.70 (95% CI: 1.88 - 17.27), 3.54 (95% CI: 1.02 - 12.36), 10.96 (95% CI: 4.74 - 25.34), 6.35 (95% CI: 1.68 - 24.05) and 11.30 (95% CI: 4.58 - 27.89) respectively. There was no first level interaction between any two risk factors. CONCLUSION: The study supported that low education level, cigarette smoking, affective conflicts, psychiatric disorders, depression were risk factors of attempted suicide and COMT Val/Val 158/108 was suspected to be a susceptible gene type of attempted suicide but needs further study. The study also suggested that 116 bp in gene atlas be possibly correlated to high activity of COMT.