Objective To analyze the VP1 and VP4 genetic region of enterovirus 71 (EV71)isolated from severe cases and mild cases with hanD-foot-mouth disease (HFMD) in Shanghai in 2011.Methods Five EV71 strains isolated from severe cases and five EV71 strains from mild cases in 2011 were included.Reverse transcription-polymerase chain reaction (RT-PCR) method was used to amplify and sequence the VP1 and VP4 genes of EV71,and then the sequencing results were compared with those of A,B,C genotype reference EV71 strains from GenBank by nucleotide alignment,amino acid alignment and phylogenetic tree analyses.Results The homogeneity between EV71 strains from severe cases and mild cases was 96.0％-98.1％ and 93.7％-99.5％ for VP1 and VP4 nucleotide sequences,respectively.The VP1 nucleotide sequences of 5 strains isolated from severe cases and 5 strains from mild cases in Shanghai shared 86.9％-98.2％ and 87.4％ 98.5％ identity with genotype C,respectively,while the homogeneity of VP4 nucleotide sequence was 85.5％-100.0％and 84.5％-99.5％,respectively.In addition,compared with the Fuyang EV71 strains (representative of C4 subtype),the strains from mild and severe cases shared homogeneity of 97.0％-98.2％ and 97.9％-98.5％ for VP1 gene,respectively,96.1％-100.0％ and 97.1％-99.5％ for VP4 gene,respectively.Among 3 strains isolated from severe cases,mutations at the residue 282 in the VP1 protein (N→S) and residue 7 in the VP4 protein (T→A) were discovered simultaneously.Conclusions The 10 EV71 strains isolated from severe and mild cases in Shanghai belong to subgenogroup C4.Among 3 strains isolated from severe cases,mutations at the residue 282 in the VP1 protein (N→S) and residue 7 in the VP4 protein(T→A) are discovered simultaneously.

Objective To test the levels of enterovirus 71 (EV71) antibody among children of different ages in Shanghai in 2011,and to investigate the relationship between antibody levels and virus infection.Methods EV71 antibody was detected by microneutralization assay from the serum specimens of healthy children of different ages collected during July to August,2011.The results were analyzed by t test for quantitative data with normal distribution,and by x2 test for count data.Results The positive rate of EV71 antibody among the 93 serum specimens was 58.1％ (54/93).The geometric mean titer (GMT) of EV71-specific neutralizing antibody was 1 ∶ 14.48.The positive rate of EV71 antibody in infants less than 6 months old was 87.5％ (21/24),and the GMT was 1∶29.56.In children aged 2 to 3 years,the positive rate of EV71 antibody decreased to 3.7％ (1/27),and GMT decreased to 1∶4.21,which were both statistically significantly lower than those less than 6 months old (x2 =36.37,t=7.58; both P＜0.01).The positive rate of EV71 antibody increased to 83.3％ (20/24) in children aged 5 to 6 years,with GMT reaching 1∶21.74.Whereas in children aged 7 to 8 years,the positive rate was 66.7％ (12/18) and GMT was 1∶20.76,without statistically significant difference compared with those aged 5 to 6 years (x2 =1.58,t=0.597; both P＞0.05).No statistically significant difference was found between boys and girls in positive rate of EV71 antibody [62.7 ％ (32/51) vs 52.4 ％ (22/42),x2 =1.02,P＞0.05] or GMT (1 ∶ 16.23 vs 1 ∶ 12.61,t=0.881,P＞0.05).Conclusions Children aged 2 to 3 years were at higher risk for EV71 infection,with EV71 antibody level significantly lower than other age groups.

Objective To understand epidemic characteristics of human influenza A and the genetic and antigenic variations of H1N1 influenza A isolates in Shanghai area in 2009. Methods Throat swabs were collected from patients with influenza-like illness in the sentinel surveillance clinic in Shanghai area in 2009, then inoculated in Madin-Darby canine kidney (MDCK) cell lines. The types of influenza were identified by direct immunofluorescence assay (DIF) and the subtypes were determined by reverse transcriptase-polymerase chain reaction (RT-PCR). Segments of hemagglutinin (HA) and neuraminidase (NA) genes of some 2009 H1N1 influenza A isolates were amplified and sequenced. HA and NA gene mutations of 2009 H1N1 influenza A isolates were analyzed. Results Seasonal H1N1 and H3N2 influenza A viruses co-circulated during the spring of 2009 in Shanghai area. Seasonal H3N2 began to co-circulate with 2009 H1N1 in August (the 32nd week) and finally2009 H1N1 became dominate since the 40th week. The phylogenetic tree of 2009 H1N1 HA segment revealed that the isolates from different regions and months were interspersed with each other, but all were clustered into one branch which closed to strains in Spain, Russia, Denmark and other European countries. Mutations were found in some HA amino acid sites, but none of them was in the antigenic determinant region. No change was observed in the 274 NA amino acid residues which were related to the drug resistance to oseltamivir. PB2 protein analysis showed that the 627 and 701 amino acid residues were glutamic acid and aspartic acid respectively, which were the same encoded amino acid with avian flu PB2 protein. Conclusions Seasonal H1N1 and H3N2 co-circulated in the spring of 2009, then both 2009 H1N1 and seasonal H3N2 were prevalent in Summer and Autumn, and 2009 H1N1 finally became dominate in Autumn. Compared to early 2009 H1N1 strains, variations are detected in H1N1 influenza A viruses, but none of them has epidemiological influence, and viruses still show high affinity with human and low-pathogenic characteristics.

ObjectiveTo investigate the immunity and seroprevalence of hepatitis A and to identify the risk factors of hepatitis A infection in 0-18 year-old children and adolescents in Shanghai.MethodsSubjects were enrolled by stratifying and clustering random sampling method.Questionnaire interview was applied to investigate the socio-demographic and behavioral factors related to hepatitis A virus (HAV),and information on HAV immunization was abstracted from the immunization registration book of each subject.The enzyme-linked immunosorbent assay (ELISA) was used to qualitatively detect HAV IgM and quantitatively measure total HAV antibody in all subjects.Risk factors associated with HAV among the subjects without HAV vaccination were analyzed.ResultsA total of 2431 subjects were enrolled in the present study with negative HAV IgM antibody and total HAV antibody in 1483 subjects were sero-positive with positivity rate of 61％.Total HAV antibody positivity rates were declined with age increasing and were significantly higher in subjects with HAV vaccination than those without HAV vaccination records.Salad food,eating together without food separation in school and endoscopy inspection were risk factors for HAV infection.ConclusionsHAV vaccination strategies remarkably improve the total HAV antibody seropositive rate in children and adolescents in Shanghai.The risk of HAV infection exists if HAV vaccination is not administrated comprehensively.Therefore,strengthening HAV vaccination and health education are important for children and adolescents to prevent and control of hepatitis A in Shanghai.

Objective To know the levels of antibodies against influenza A virus subtypes H1 and H3 of population in Shanghai during 2009, and the detection of antibodies against avian influenza virus subtypes H5 and H9 in population which contacts with avian. Methods The serological survey of the antibodies against influenza A viruses subtypes H1, H3, H5 and H9 in 356 close contacts with avian (professional population) and 332 general subjects (general population) at various age groups were carried out using hemagglutinin inhibit (HI) test. Results The positive rates of antibodies against influenza virus A/Brisbane/59/2007 (H1N1) in general population and professional population were 82.8% (275/332) and 73.9% (263/356), respectively; those of A/Brisbane/10/2007 (H3N2)were 50.6% (168/332) and 54.8% (195/356), respectively. The positive rate of antibodies against influenza virus A/Brisbane/59/2007 (H1N1 )was significantly higher than that of influenza A viruses subtype H3, which was consistent with etiological survey of influenza virus in Shanghai during 2008.The positive rates of antibodies against influenza A virus subtype H5 in professional population and general population were 4.2% (15/356) and 0.3% (1/332), respectively; those of influenza A virus subtype H9 were 34.6% (123/356) and 2.4% (8/332), respectively. The positive rates of antibodies against influenza virus A/Brisbane/59/2007 (H1N1 ) and A/Brisbane/10/2007 (H3N2) in age groups of 6 months-5 years and ≥60 years were lower than other age groups. Conclusions The immune protective response against seasonal influenza A subtype H1 and H3 of population in Shanghai is high,while those of children and the elders were low. The levels of antibodies against influenza A viruses subtype H5 and H9 in professinal population present obviously ascending trend, which indicates that the etiological and serological survey of influenza virus in this population should be enhanced.

OBJECTIVEProtein induced by vitamin K absence or antagonist II (PIVKA II), also called des-gamma carboxy prothrombin (DCP), is a sensitive marker for the diagnosis of hepatocellular carcinoma (HCC), in Japan and the United States since the sensitive kits were available (1998). PIVKA II is not used in clinical diagnosis in China so far. The aim of this study was to assess the diagnostic value of PIVKA II in Chinese patients with HCC.

METHODSSerum PIVKA II and alpha-fetoprotein (AFP) levels were determined in 60 patients with HCC and 30 patients with cirrhosis not carrying HCC.

RESULTSThe mean serum concentration of PIVKA II in HCC patients (784.3 +/- 1364.1 mean +/- s) was higher than that in cirrhosis patients (16.1 +/- 31.7); this difference was highly significant (P < 0.0001). When the cutoff level of 40 mAU/ml was used as the level of discriminating HCC from cirrhosis, 51.7% of patients (31/60) with HCC had PIVKA II values above this level (sensitivity). Only 4 patients with cirrhosis had such high PIVKA II levels. Thus, the specificity of this test was 86.7% (26/30). Total accuracy was 62.2% [(31 + 26)/(60 + 30)]. Seven of 19 small HCCs (36.84%) had PIVKA II values above the cutoff level. Concentrations of AFP above 20 ng/ml were observed in 34 of 60 patients with HCC (56.7%) and in 11 patients with cirrhosis (36.7%). Eleven of 26 patients with HCC (46.2%) without increased AFP had concentrations of PIVKA II greater than 40 mAU/ml. No significant correlation was found between serum levels of AFP and PIVKA II that were measured in 60 HCC patients (rs = 0.101, P = 0.247). Combining the information from PIVKA II and AFP showed an increase of approximately 21.6% over AFP and 26.7% over PIVKA II alone. For small HCC patients, combining the information from PIVKA II and AFP showed an increase of approximately 15.8% over AFP alone and 21.1% over PIVKA II alone.

CONCLUSIONPIVKA II is a useful early diagnostic marker for HCC and may be more sensitive when combined with AFP in Chinese patients.

Adult ; Aged ; Aged, 80 and over ; Biomarkers ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; blood ; diagnosis ; pathology ; Female ; Humans ; Liver Cirrhosis ; blood ; Liver Neoplasms ; blood ; diagnosis ; pathology ; Male ; Middle Aged ; Protein Precursors ; blood ; Prothrombin ; alpha-Fetoproteins ; analysis