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Canine distemper virus (CDV) causes a highly contagious disease in a wide range of carnivores. The hemagglutinin (H) protein of viruses shows the highest variability and plays an important role in modulation of viral antigenicity, virulence, and receptor recognition. Since 2012, canine distemper (CD) outbreaks in fur-bearing animals (minks, foxes, raccoon dogs) caused by CDV variants with I542N and Y549H substitutions in the H protein have been frequently reported in China. To characterize the molecular evolutionary dynamics and epidemiological dynamics of CDV, 235 H gene sequences of CDV wild-type strains collected from 22 countries between 1975 and 2015, including 44 strains predominant in fur-bearing animals in China, were analyzed. The phylogenetic relationships and evolutionary rates of the CDV strains were determined by Bayesian phylogenetics. The CDV strains clustered into distinct geographic genotypes, irrespective of the species of isolation. All the variant strains formed a distinct monophyletic cluster and belonged to the F sub-genotype within the Asia-1 genotype-currently the predominant sub-genotype in fur-bearing animals in China. Evolutionary analysis suggested that the variant strains originated in 2006. Furthermore, the selection pressure analysis revealed that the Y549H substitution was under positive selection pressure for adaptation toward the fur-bearing animals. The residue at position 549 also showed structural interaction with the V domain of the mink signaling lymphocyte-activation molecule (SLAM) receptor based on the homology modeling of the H-SLAM complex. Our results suggested that the Y549H substitution contributed to the molecular adaptation of CDV variants in the fur-bearing animals during the viral evolutionary phase in China.

To investigate the expression and biological significance of LINC00310 in the malignant transformation of human bronchial epithelial cells(16 HBE) induced by glycidyl methacrylate(GMA).The 16 HBE cells recovered successfully used 1 μg/mL dimethyl sulfoxide(DMSO) as the solvent control group, and the final concentration was 8 μg/mL GMA as the treatment group, and were subcultured after repeated exposure 3 times for 72 hours each time. The 10 th, 20 th and 30 th generation cells of the GMA treatment group and corresponding DMSO control group were collected. The LncRNA microarrays was used to analyze the expression changes of LINC00310 in different periods, and the target gene and function prediction was performed by NCBI and cBioPortal bioinformatics database, and real-time quantification PCR(qPCR) was used to detect the relative expression levels of LINC00310 and predicted target genes.The result of the microarray showed that LINC00310 in the GMA-treated group was down-regulated by 2. 02-fold, up-regulated by 6. 17-fold, and up-regulated by 2. 03-fold in the pre-transformation, mid-term, and late, respectively. The result of qPCR confirmed that the expression of LINC00310 relative expression level of 10 th, 20 th and 30 th generation cells was consistent with the microarray result, which were down-regulated by 2. 76-fold, up-regulated by 2. 68-fold, and up-regulated by 3. 09-fold. Consistently, the relative expression of the target gene C-Myc was statistically significant in 20 th and 30 th generation cells.LINC00310 induced low expression in the early stage of malignant transformation of 16 HBE cells induced by GMA, and was highly expressed in the middle and late stages. It indicated that LINC00310 may play a cancer-promoting role in the process of cell malignant transformation through C-Myc.

α-Toxin, a pore-forming toxin secreted by most Staphylococcus aureus, plays critical role in the pathogenesis associated with various infectious diseases. The USA300 which is a major international epidemic methicilin-resisrant S. aureus has spread rapidly to multiple countries and become an emerging public health concern. In this study, the in vitro efficacy of Dracorhodin Perochlorate (DP) against USA300 virulence was evaluated. Using susceptibility testing, immunoblots, rabbit blood haemolytic assay and real-time RT-PCR, we observed that the α-toxin production was decreased when USA300 was co-cultured with different sub-inhibitory concentration of DP. Further, the protective effect of DP against USA300-mediated injury of human alveolar epithelial cells (A549) and MH-S cells was evaluated by cytotoxicity assays, and the result revealed that DP, at final concentration of 16 µg/ml, is a potent antagonist for USA300-mediated cell damage. Importantly, those beneficial effects might partially correlate with hla and RNAIII suppression by DP, leading to the inhibition of α-toxin production in culture supernatant. Overall, these results suggest that DP could attenuate the virulence of USA300 by decreasing α-toxin production without inhibiting bacterial growth, and this compound may represent an ideal candidate for the development of anti-virulence agent combating S. aureus infection.

To analyze the opioid binding protein/cell adhesion molecule-like (OPCML) methylation status at different stages of malignant transformation of human bronchial epithelial (16HBE) cells induced by Glycidyl Methacrylate (GMA) and to explore the effect of OPCML methylation in the process of malignant transformation.Cells were harvested at different stages (the 10th generation, the 20th generation and the 30th generation). To verify the Methylation chip result of OPCML methylation status in the process of malignant transformation, we detected it by methylation-specific-PCR (MSP); Real-time fluorescence Quantitative PCR (qPCR) were applied to measure the gene expression levels of OPCML at different transformed stage, and compared with the control groups (treated with DMSO).Based on the result of methylation chip, the gene of OPCML methylation occurred in all stages, which was consistent to the result of MSP; qPCR showed that the levels of gene expression decreased in the 20th generation and 30th generation.Methylation status of OPCML gene promoter could be considered as a stable and specific biomarker in the transformation process.

Background Bovine tuberculosis has led to serious economic losses for Sika Deer producers in China. Strategies for controlling the spread of Mycobacterium bovis are often hampered by a lack of epidemiological data. Specifically, tracing infections requires the ability to trace back infections, which, in turn, requires the ability to determine isolates with a common source. This study was planned to assess the discriminatory power of each mycobacterial interspersed repetitive unit (MIRU)-variable number tandem repeats (VNTR) locus and evaluate the most appropriate combination of MIRU-VNTR loci for molecular epidemiological studies on Sika Deer in China. Results The discriminatory power of MIRU-VNTR typing based on 22 known loci (12 MIRUs, 2 ETRs, 4 QUBs, and 4 Mtubs) were assessed in 96 Mycobacterium bovis strains collected sequentially from Sika Deer at a slaughterhouse in northeastern China. We defined four loci (MIRU4, ETRA, QUB11b, and Mtub4) as highly discriminative, eight loci (MIRU2, MIRU23, MIRU27, MIRU31, MIRU39, MIRU40, QUB26, and Mtub21) as moderately discriminative, and three loci (MIRU16, Mtub30, and Mtub34) as poorly discriminative. The final locus showed no polymorphism between strains. MIRU-VNTR typing as a whole was highly discriminative, with an overall allelic diversity of 0.897. Of the loci tested, the four highly discriminative loci and eight moderately discriminative loci proved to be most appropriate for first line typing of M. bovis from Sika Deer, with the same resolving ability as all 22 loci (H = 0.897). Conclusions MIRU-VNTR typing is quick and effective for typing bovine tuberculosis isolates from Sika Deer in China. Electronic supplementary material The online version of this article (doi:10.1186/s12917-015-0402-0) contains supplementary material, which is available to authorized users.

To develop a method for determination of 2, 4-dichlorophenoxyacetic acid (2, 4-D) in the air of workplace by high-performance liquid chromatography.2, 4-D was collected by ultrafine glass filters, desorbed by methanol, separated by a C18 column, and detected by a UV detector. Identification and quantification of 2, 4-D were performed by retention time and peak areas, respectively.The linear range of the test was 2∼200 µg/ml; the elution efficiency was 94.6%- 95.9%; the limit of detection (S/N = 3) was 0.034 µg/ml (injection volume of 20 µl eluant); the lower limit of quantification (S/N = 10) was 0.11 µg/ml; the minimum detectable concentration was 0.011 mg/m(3); the minimum quantifiable concentration was 0.037 mg/m(3) (with sampled air volume of 45 L).This method is convenient and simple in sample collection and preparation, and satisfies all methodological requirements. Therefore, this method is useful for the determination of 2, 4-D in the air of workplace.

Three novel metabolites (1, 5 and 10) were isolated from a 25-L fermentation broth of Actinomyces sp. JN411010 together with 11 related compounds. Their structures were determined on the basis of 1D and 2D NMR, as well as HR-ESI-MS spectroscopic data analyses. Compounds 1-2, 5-8 and 10 displayed moderate antimicrobial activities against Gram-positive and Gram-negative bacteria. Biosynthetic pathway of 5 and 10 was proposed to be catalysed by polyketide and non-ribosomal peptide gene clusters, respectively.

To observed the chromosome oberration of CHL cells induced by CdTe QDs.The chromosome oberration test of Chinese hamster lung (CHL) cell was conducted in CdTe QDs at the concentrations of 1.6, 3.2, 6.3, 12.6 and 25.2 microl/ml, under the metabolic (+S9) and non-metabolic (-S9) activation systems.The chromosome oberration rate of 12.6 and 25.2 microl/ml CdTe QDs groups were significantly higher than the control groups (P0.01) Under the condition of metabolic activation. Main types of chromosomal aberrations was broken, fragmented and exchange.It was suggested that CdTe QDs could induce the effects of chromosome oberration under the metabolic activation systems (+S9).

Eukaryotic expression plasmid (PVAX 1/E0) of E0 gene was constructed from an isolated strain of bovine viral diarrhea virus (BVDV) from sika deer and transfected into the BHK-21 cells to detect E0 gene transcription expression level and protein expression level. It was found that E0 gene could express in BHK-21 cells by RT-PCR and ELESA results. The humoral and cellular immune response levels were detected in rabbits immunized with PAVX1/E0 and inactivated vaccine by ELESA technology, which reached the peak after 42 days immunization and PVAX1/E0 immune rabbits could produce specific higher cells immune responses level in comparison to inactivated vaccine group. This is the first report demonstrating and “immunogenicity” of BVDV E0 gene on “sika deer”, indicating a new vaccine target for BVDV. Key words: Sika deer, bovine viral diarrhea virus, E0 gene, eukaryotic expression, immune response.

To explore the correlation between genetic polymorphisms of XRCC1, XPD, XRCC3 and DNA repair capacity induced by benzene.Eighty patients suffered from chronic benzene poisoning were investigated. PCR-RFLP was applied to detect the single nucleotide polymorphisms on C26304T, G27466A, G28152A, G36189A of XRCC1, C22541A, C23591T, A35931C of XPD, C18067T of XRCC3. Cytokinesis-block micronucleus (CBMN) and alkaline comet were applied to detect the DNA repair capacity.The DNA repair capacity of the subjects carrying XPD 35931C variant allele or carrying XRCC3 18067 C/T variant genotype were higher than those carrying corresponding mild genotype.There could be a correlation between polymorphisms of XRCC3 and DNA repair capacity of DNA damage induced by benzene.

To investigate the effect of glycidyl methacrylate (GMA) on cell cycle and apoptosis in human bronchial epithelial cells.Flow cytometry was applied to determine cell cycle and apoptosis of 16HBE cells treated at the doses of 1-16 microg/ml GMA for different time. Mitotic index (MI) of cell was also analysed.When 16HBE cells were treated by GMA for 72hr, cells in G0/G1 phase decreased, but cells in S and G2/M phase increased. The increases of apoptosis rates and the decreases of mitotic index could also be observed. As treatment went on, all groups cells were stopped at G0/G1 phase, but at the dose of 16 microg/ml the cell increases of S and G2/M phase were observed in comparison with the controls. After cells were treated for 3 times, the decreases apoptosis rates and the increases of mitotic index of cell treated by high dosage were observed.The 16HBE cells treated by GMA could result in cell cycle transition from G0/G1 phase to S and G2/M phase. It was suggested that the cell proliferation was stimulated. Uncontrolled cellular proliferation may be responsible for malignant transformation induced by GMA.

Background Bovine viral diarrhea virus (BVDV) infections continue to cause significantly losses in the deer population. Better isolation and identification of BVDV from sika deer may contribute significantly to the development of prophylactic therapeutic, and diagnostic reagents as well as help in prevention and control of BVDV. However, isolation and identification of BVDV from sika deer is seldom reported in literature. In this study, we collected some samples according to clinical sign of BVDV to isolation and identification of BVDV from sika deer. Results we isolated a suspected BVDV strain from livers of an aborted fetus from sika deer in Changchun (China) using MDBK cell lines, named as CCSYD strain, and identified it by cytopathic effect (CPE), indirect immunoperoxidase test (IPX) and electron microscopy(EM). The results indicated that this virus was BVDV by a series of identification. The structural proteins E0 gene was cloned and sequenced. The obtained E0 gene sequence has been submitted to GenBank with the accession number: FJ555203. Alignment with other 9 strains of BVDV, 7 strains of classical swine fever virus (CSFV) and 3 strains of border disease virus(BDV) in the world, showed that the homology were 98.6%-84.8%, 76.0%-74.7%, 76.6%-77.0% for nucleotide sequence, respectively. The phylogenetic analysis indicated that new isolation and identification CCSYD strain belonged to BVDV1b. Conclusion To the best of our knowledge, this is the first report that BVDV was isolated and identified in sika deer. This current research contributes development new BVDV vaccine to prevent and control of BVD in sika deer.