Objective: To analyse the genomic characteristics of Novirus(NoV) GII.4 genotype JN010 strain isolated form Jinan in 2017. Methods: Seven pairs of primers were designed and used to amplify the JN010 genome. Sequence analyses, alignment and phylogenetic trees of ORF1 and ORF2 genes were performed using the software Lasergene7.1 and MEGA5.2. At the same time the major protein VP1 amino acid mutations were analyzed. Results: The 7 516 bp complete genome sequence of JN010 strain was obtained, the most mutation sites were located in P2 subdomain of VP1. Two substitutions I293N and H373N of VP1 were locate neighboring epitope A, and R297H mutation happened within epitope A and the site A that binding with histo-blood group antigens(HBGAs). The JN010 strain was GII.Pe/GII.4 genotype and genetically closest to the strains found in Osaka of Japan(GenBank accession number LC066046) and the strains in Zhongshan city of Guangdong (GenBank accession number KY407064) respectively according to ORF1 and ORF2 gene homologous and phylogenetic analysis. Conclusions The NoV GII.4 variant strain JN010 has occurred mutations in the key site of the epitope A and site A that bind with HBGAs, and maybe affect its antigenicity and interaction with HBGAs.

Objective:To elucidate the changes of cytokine expression profiles in hand, foot and mouth disease (HFMD) patients infected with non-EV-A71 enteroviruses in Jinan city, and explore the characteristics of cytokines expression.Methods:The serum samples of acute and convalescent phases were collected from non-EV-A71 enterovirus-infected HFMD patients in Jinan from 2014 to 2017. The serum samples of healthy subjects were collected as control group. The Bio-plex liquid chip platform was used for high-throughput detection of 27 cytokines. GraphPad Prism and SPSS 22.0 were used for description and statistical analysis.Results:Twenty-two serum cytokines significantly changed in non-EV-A71 infected patients, including 11 kinds of interleukin (IL), 5 kinds of chemokines, 2 kinds of growth factors, granulocyte colony stimulating factor (G-CSF), interferon-γ, tumor necrosis factor-α (TNF-α) and granulocyte-macrophage colony stimulating factor (GM-CSF). There were 21 kinds (mean ranks 17.06-19.00 pg/ml vs 5.50-8.80 pg/ml, P < 0.05) and 20 kinds (mean ranks 16.41-19.00 pg/ml vs 5.50-9.90 pg/ml, P < 0.05) of cytokines expression in acute stage and convalescent stage respectively were higher than those in healthy control group for coxsackievirus A16 (CV-A16) infected patients, and GM-CSF expression (mean ranks 9.65 pg/ml vs 21.40 pg/ml, 9.59 pg/ml vs 21.50 pg/ml, P < 0.05) were both lower than those in healthy control group. For HFMD patients infected CV-A6, there were 19 kinds (mean ranks 11.92-13.50 pg/ml vs 5.50-6.45 pg/ml, P < 0.05) and 21 kinds (mean ranks 12.00-13.50 pg/ml vs 5.50-6.40pg/ml, P < 0.05) of cytokines expression with acute and convalescent stage respectively were higher than those in healthy control group. GM-CSF expression decreased only in acute phase (mean ranks 5.00 pg/ml vs 10.60 pg/ml, P < 0.05) compared with healthy control group. Double serum analysis showed that interleukin 6 (22.79pg/ml vs 35.88 pg/ml) and interferon-induced protein 10 (IFNγ -induced protein 10) (793.56 pg/ml vs 2 157.32 pg/ml) expression in patients with CV-A16 infection during convalescent stage were lower than that in acute stage; IL-7 (3.13 pg/ml vs 1.165 pg/ml), IL-15 (27.84 pg/ml vs 16.005 pg/ml) and regulated upon activation normal T cell expressed and secreted (RANTES) (22 605.96 pg/ml vs 7 040.90 pg/ml) expression in patients with CV-A6 infection during convalescent stage increased compared with the acute stage. Conclusions:There are extensive changes in cytokine expression profile in HFMD patients with non-EV-A71 enterovirus infection. Different pathogens infection and different clinical course of HFMD have different characteristics of cytokine expression. These findings could provide scientific data for finding indicators that are meaningful for disease progression, clinical diagnosis and immunotherapy.

Objective To detect and analyze the levels of 27 serum cytokines expression in EV71-induced Hand Foot and Mouth Disease(HFMD) in Jinan and revealed their correlations with disease severity based on the platform of liquid chip.Methods 43 serum samples were collected from EV71-infected HFMD patients in Jinan in 2009-2014,including 16 mild cases and 27 severe cases.10 serum specimens from healthy people were also collected as controls.27 serum cytokines were analyzed on Bio-Plex liquid chip platform.Results Compared to healthy controls,22 cytokines expression levels increased in severe groups,including IL-1 ra,IL-2,IL-4,IL-5,IL-6,IL-7,IL-8,IL-9,IL-10,IL-12 (p70),IL-13,IL-15,IL-17,etotaxin,G-CSF,IFN-γ,IP-10,MCP-1,PDGF-BB,MIP-1β,RANTES and TNF-α(P ＜ 0.02).21 cytokines expression levels elevated than healthy controls (P ＜ 0.02).GM-CSF was found decreased levels in both mild and severe groups than in the healthy controls(P ＜ 0.02).The correlation analysis showed that 12 cytokines (IL-1 ra,IL-7,IL-9,IL-10,IL-13,IL-17,etotaxin,IP-10,PDGF-BB,MIP-1β,RANTES and TNF-α) were moderately correlated with the severity of EV71-infected HFMD.GM-CSF was negatively correlated with HFMD.Conclusions Many kinds of serum cytokine changed obviously in the EV71-infected HFMD patients and cytokine levels were closely associated with the severity of HFMD.The analysis of liquid chip for serum cytokines provides an efficiently method.

Objective:Serum cytokines and chemokines in patients with fever with thrombocytopenia syndrome (SFTS) and the severity of the disease was studied.Methods:Serum of 47 SFTS patients from 2014 to 2017 in Jinan city was collected. Serum of 11 dead patients and 36 survivors were selected as the survival group and serum of 10 healthy people was selected as the control group. Luminex liquid phase chip was used to detect and analyze the expression levels of 40 cytokines and chemokines in peripheral blood.Results:The expression levels of 27 cytokines and chemokines in the death group were significantly higher than those in the control group ( P<0.05). The expression level of 33 cytokines and chemokines in the death group was significantly higher than that in the survival group ( P<0.05). The expression level of 12 cytokines and chemokines in the survival group was significantly higher than that in the control group ( P<0.05). The expression levels of 10 cytokines and chemokines in the patients were increased in different degrees during the recovery period compared with the acute period ( P<0.05). The expression levels of 5 cytokines and chemokines were lower in the recovery period than in the acute period ( P<0.05). Receiver operating characteristic curve (ROC curve) analysis showed that 22 factors were statistically significant and could be used as biomarkers to predict the severity of the disease. Conclusions:The expression levels of various cytokines and chemokines in serum of SFTS patients changed significantly during the development of the disease and the cytokine storm caused by SFTS may be the cause of death.

Objective: To analyze the VP1 gene characteristics of Coxsackievirus A16 (CV-A16) isolated in(hand, foot and mouth disease, HFMD) patients of Jinan 2017. Methods: The samples collected from patients with HFMD in Jinan city in 2017 were analyzed and some of the CV-A16 nucleic acid positive samples were choosen with simple random sampling method and pretreated to inoculate RD cells. The whole VP1 gene sequences of CV-A16 stains which had obvious cytopathic effect in RD cells were amplified and sequenced. MEGA5.2 software was used to constructed phylogenetic tree and Lasergene software was used to analysis the homology consistency of nucleotide and amino acid of VP1 gene. Results: All the 10 CV-A16 strains isolated from Jinan were located 3 clusters belonged to B1b genotypes, with a nucleotide similarity of 94.9%~100% and deduced amino acid similarity of 99.3%~100%. The Jinan CV-A16 strains were nearest related with Shanghai strains KX871333(nucleotide similarity: 99.7%), Henan strains KM260134(99.0%), Jiangsu strains KP751580、KR138327(98.7%) and Guangdong strains MH004016(98.5%). Compared with some reference strains from our country, amino acid mutation were identified at positions 14, 59, and 145 in VP1 proteins of Jinan CV-A16 strains. Conclusions CV-A16 strains B1b genotypes were the strains prevailed in Jinan city, and 3 amino acid substitutions in VP1 proteins were happened in CV-A16 strains isolated from Jinan.