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We wished to analyze the genetic characterization of echovirus 11 (Echo11) from samples of environmental sewage in Shandong Province (China). The VP1 coding region was typed as the strains were amplified. Phylogenetic analyses on the VP1 sequences from these isolates, strains isolated from AFP cases in the period 1994-2010 and others published in GenBank were conducted. From 2011 to 2012, 94 Echo11 strains were isolated from samples of environmental sewage in Jinan and Linyi City in Shandong Province. Numbers of Echo11 were seasonal and reached peaks in the summer and autumn in both cities; A- mong these isolates, nucleotide (nt) identities were 89.5%-100.0% whereas amino acid (aa) identities were 95.4%-100.0%. The nt and aa identities were 76.6%-79.7% and 90.4%-92.5% between those strains and the prototype (Gregory) strain of Echo11, respectively. All isolates from Shandong Province were the A genotype and the strains evolved very rapidly, which suggested that several transmission chains was co-circulating. We described the temporal fluctuation and genetic characterization of Echo11 isolates from surveillance of environmental sewage in Shandong Province, thereby providing important information for exploring the dynamic change and genetic variation of circulating human enteroviruses in this Province in China.

Human Enterovirus HEV 74 is a new member of species Human enterovirus B (HEV-B). To understand its evolution and restructuring characteristics, we report the complete genome sequence of a HEV74 strain 05293/SD/CHN/2005(abbreviated as 05293) isolated from an acute flaccid paralysis (AFP) case in Shangdong Province, China, 2005. Analysis of the complete genomic sequence of 05293 showed that its genome was collinear with that of previously described 2 HEV74 strains, except for insertions and deletions at the 5'NTR and the 3 NTR regions. The complete genome sequence of strain 05293 displayed 80. 8% nucleotide and 96% amino acid identity to the prototype strain USA/CA75-10213, and 80. 6% and 95. 9% to another isolated strain Rikaze-136. The P1, P2 and P3 coding regions of strain 05293 displayed 81. 5%, 80. 0%, 79. 7% nucleotide and 95. 9%, 96. 0%, 96.2% amino acid identity to the prototype strain USA/CA75-10213, and 81. 9%, 78. 8%, 79. 5% and 95. 9%, 96. 1%, 95. 7% to strain Rikaze-136, respectively. The phylogenetic tree and Simplot analysis on 05293 and HEV-B genome sequences were performed, and the result indicated frequent recombination within HEV-B.

Objective: To investigate the pathogen spectrum constitution of HEV related to acute meningitis/encephalitis syndrome (AMES) and to analyze the genetic characterization of the echovirus 30 (Echo30) isolates., Methods: Cerebrospinal fluid, stool, and/or throat swab specimens from 101 AMES cases were collected for virus isolation with RD and HEp-2 cell lines in Linyi city from April to December, 2010. After typing by neutralization test, the entire VP1 gene of the isolates were amplified by RT-PCR and sequenced. Homologous comparison was carried out and phylogenetic analysis were performed., Results: HEVs were isolated from 47 specimens of 34 patients (4 from cerebrospinal fluid, 18 from throut swab and 25 from stool), with 11 Echo30 isolates identified by microneutralization assay and molecular typing method. In alignment of VP1 sequences with global Echo30 isolates, relative high nucleotide homologies (91.2% - 95.4%) with Taian, Zhangqiu and Jiangsu isolates and high divergences (17.9% -19.6%) with prototype strain were observed. Nucleotide divergences among Linyi isolates were 0 - 10.2%, and 3 lineages were revealed via phylogenetic analysis, reflecting 3 transmission chains of Echo30 co-circulated in Linyi city, 2010., Conclusion: Echo30 was the predominant serotype of AMES cases in Linyi city, 2010. These isolates possessed considerable divergence with Echo30 from other countries.

Objective: To study the protective effects of tetramethylpyrazine (TMPZ) on rat myocardial cells infected by Coxsackie virus B3 (CVB3) and its signal transduction mechanism., Methods: The cultured myocardial cells of neonatal Sprague-Dawley rats were randomly treated with CVB3, CVB3+TMPZ (100 micromol/L), TMPZ (100 micromol/L) (negative control) or DMEM (blank control). After treatment, the beating rate of myocardial cells and the LDH activity in the culture fluid were measured. Cell viability was ascertained with MTT assay. Western blot was used to study the expression of nuclear factor kappa-B (NF-kappaB) protein in myocardial cells., Results: The beating rate of myocardial cells in the untreated CVB3 infection group was significantly lower than that in the TMPZ-treated CVB3 infection group (32.0+/-3.6 bpm vs 84.3+/-3.5 bpm, P<0.01). The LDH activity and NF-kappaB expression in the TMPZ-treated CVB3 infection group was significantly reduced when compared with untreated CVB3 infection group (P<0.05, P<0.01, respectively). Cell viability 7 days after CVB3 infection in the TMPZ-treated group was higher than that in the untreated CVB3 infection group (86.7+/-2.7% vs 35.3+/-3.4%; P<0.01)., Conclusions: TMPZ can provide protective effects on rat myocardial cells infected by CVB3, possibly by an inhibition of the activity of NF-kappaB in myocardial cells.