Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis in Asia and domestic pigs serve as the amplifying hosts. In the present study, the full genomic sequences of two JEV strains (HEN0701 and SH0601) isolated from pigs in China were determined and compared with other 12 JEV strains deposited in GenBank. These two strains had an 88.8% nucleotide sequence similarity and 97.9% deduced amino acid sequence homology. HEN0701 had high nucleotide sequence and high amino acid sequence identity with genotype I (GI) strains, while SH0601 had high nucleotide sequence and high amino acid sequence identity with GIII strains at both the gene and full genome levels. Further phylogenetic analysis showed that HEN0701 belonged to the JEV GI group and SH0601 was classified as a GIII strain. Analysis of codon usage showed there were a few differences between the GI and GIII strains in nucleotide composition and codon usage for the open reading frames.
Animals ; Cell Line ; Cricetinae ; Encephalitis Virus, Japanese/classification/*genetics ; Encephalitis, Japanese/epidemiology/*veterinary/virology ; Gene Expression Regulation, Viral/physiology ; Genome, Viral ; Molecular Epidemiology ; Phylogeny ; Swine ; Swine Diseases/epidemiology/*virology

Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis in Asia and domestic pigs serve as the amplifying hosts. In the present study, the full genomic sequences of two JEV strains (HEN0701 and SH0601) isolated from pigs in China were determined and compared with other 12 JEV strains deposited in GenBank. These two strains had an 88.8% nucleotide sequence similarity and 97.9% deduced amino acid sequence homology. HEN0701 had high nucleotide sequence and high amino acid sequence identity with genotype I (GI) strains, while SH0601 had high nucleotide sequence and high amino acid sequence identity with GIII strains at both the gene and full genome levels. Further phylogenetic analysis showed that HEN0701 belonged to the JEV GI group and SH0601 was classified as a GIII strain. Analysis of codon usage showed there were a few differences between the GI and GIII strains in nucleotide composition and codon usage for the open reading frames.
Animals ; Cell Line ; Cricetinae ; Encephalitis Virus, Japanese/classification/*genetics ; Encephalitis, Japanese/epidemiology/*veterinary/virology ; Gene Expression Regulation, Viral/physiology ; Genome, Viral ; Molecular Epidemiology ; Phylogeny ; Swine ; Swine Diseases/epidemiology/*virology

OBJECTIVETo study the effect of annexin A5 on the apoptosis of laryngeal cancer cells.

METHODSSpecial siRNAs were used to knock annexinA5 down in Hep-2 cell, and RT-PCR and Western blot were applied to identify the efficacy of RNA interference. The flow cytometry assay was performed to detect the Hep-2 cell apoptosis.

RESULTSRT-PCR analysis showed that the relative mRNA expression of annexin A5 in siRNA group, negative control group, Lipofectamine 2000 group and blank control group were 0.70 ± 0.03, 1.18 ± 0.05, 1.17 ± 0.06 and 1.23 ± 0.07. The relative mRNA expression of annexin A5 in siRNA group was significantly decreased than contrast groups(t = -14.77, t = -13.23, t = -12.99, P < 0.05).In Western blot assay, the trend of protein expression level was consistent with the mRNA expression levels of annexin A5. The relative levels of proteins in siRNA group, negative control group, Lipofectamine 2000 group and blank control group were shown 1.21 ± 0.03, 3.88 ± 0.06, 3.87 ± 0.02 and 3.95 ± 0.08. The relative protein expression of annexin A5 in siRNA group was significantly decreased than contrast groups(t = -70.34, t = -150.62, t = -56.32, P < 0.05). At the same time in flow cytometry the apoptotic rate of siRNA group, negative control group, Lipofectamine 2000 group and blank control group were 4.43% ± 0.12%, 13.67% ± 0.22%, 13.66% ± 0.12% and 13.35% ± 0.13%, the difference between the siRNA group and contrast groups was statistically significant(t = -62.50, t = -14.16, t = -11.47, P < 0.05).So after RNA interference, expression of annexin A5 decreased, and the results in the apoptosis inhibition of Hep-2 cell.

CONCLUSIONAnnexin A5 promotes apoptosis of Hep-2 cells, and it may be a potential therapeutic target for the laryngeal cancer.

Annexin A5 ; genetics ; Apoptosis ; genetics ; Cell Line, Tumor ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering