In order to understand the management measures,technique abilities and difficulties in controlling and eradicating avian diseases in parent breeding flocks,the current situation of avian disease eradication in breeding flocks was investigated by China Animal Disease Control Centre in 19 provinces in July 2015.Questionnaires investigation was conducted and the feedbacks were received from 214 parent breeding flocks.This study summarized and analyzed the information of farm management,breeding resources,avian disease surveillance and disease eradication in these flocks,which will provide the basic data to promote the avian diseases control and eradication in China.

Objective To assess the effect of silent mating type information regulation 2 homolog 6 (SIRT6) on migratory and proliferative activity of skin fibroblasts in the elderly,and to explore their mechanisms.Methods Circumcised foreskins were obtained from patients of different ages in Department of Urologic Surgery,Second Hospital of Shanxi Medical University,including 8 elderly patients and 8 young patients.Human skin fibroblasts were isolated from the foreskin tissues by using a collagenase digestion method.Western blot analysis was performed to determine the expression of SIRT6 and phosphorylated p65 (p-p65) in human skin fibroblasts in different age groups,scratch assay to evaluate cell migratory activity,and cell counting kit-8 (CCK8) assay to assess cellular proliferative activity.Skin fibroblasts in the elderly group were divided into 2 groups:SIRT6 group transfected with a lentiviral vector overexpressing SIRT6,and control group transfected with an empty lentiviral vector.Then,the cell migratory and proliferative activity as well as p-p65 expression were measured by the above methods,and the mRNA expression of type Ⅰ and Ⅲ collagens,and integrin subunits α3,α5 and β1 was determined by real-time PCR in the SIRT6 group and control group.Statistical analysis was carried out with GraphPad Prism 5 software by using t test for comparison between 2 groups.Results Compared with the young group,the elderly group showed significantly decreased SIRT6 expression in skin fibroblasts (0.434 ±0.179 vs.1.000 ± 0.067,t =3.040,P =0.012),migration rate (43.81％ ± 18.84％ vs.94.63％ ± 12.32％,t =5.903,P =0.003)and cellular proliferative activity at 24 and 48 hours (both P ＜ 0.05),but significantly increased p-p65 expression (1.694 ± 0.148 vs.1.000 ± 0.093,t =2.949,P =0.015).Compared with the control group,the SIRT6 group showed significantly decreased p-p65 expression (P ＜ 0.05),but significantly increased migratory and proliferative activity (both P ＜ 0.05),and elevated mRNA expression of type Ⅲ collagen and integrin subunits oα3,α5 and β1 (all P ＜ 0.05).Conclusion SIRT6 can improve the migratory and proliferative activity of human fibroblasts in the elderly,possibly by inhibiting the nuclear factor-κB pathway.

Porcine reproductive and respiratory syndrome virus (PRRSV) non-structural protein 7 (Nsp7) plays an important role in the induction of host humoral immune response and could serve as an ideal antigen for serological genotyping assay for PRRSV based on the significant difference in immunoreactivities of North American (NA) and European (EU) PRRSV Nsp7. In this study, Nsp7 of NA and EU PRRSVwas separately expressed and purified using prokaryotic expression system. The purified recombinant Nsp7 proteins reacted with serum antibodies against corresponding genotype PRRSV in Western blotting. However, nonspecific reaction of whole recombinant Nsp7 with antibodies against another genotype PRRSV was observed, indicating that whole NA PRRSV Nsp7 and EU PRRSV Nsp7 have similar antigenic epitopes and recombinant proteins could not be used for genotyping of antibodies against PRRSV. Based on the analysis of similar antigenic epitopes at the hydrophilic region of NA PRRSV Nsp7 and EU PRRSV Nsp7 by bioinformatics assessment, partial Nsp7 gene region deleted sequences encoding similar antigenic epitopes was constructed by fusion PCR. The recombinant truncated Nsp7 (NA-deltaNsp7 and EU-deltaNsp7, about 43 kDa) was expressed and the molecular weight was about 43 kDa. The results of Western blotting showed that NA-deltaNSP7 and EU-deltaNSP7 could be specifically recognized by positive serum to NA or EU PRRSV individually and nonspecific reaction was eliminated. This study provided a basis for further development of serological genotyping assay for North American and European genotype PRRSV infection.
Animals ; Genotype ; Porcine respiratory and reproductive syndrome virus ; classification ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; immunology ; Swine ; Viral Nonstructural Proteins ; biosynthesis ; immunology