ABSTRACT
Objective To analyze differences in gene expression profile of pituitary tissue in cervical spondylosis of vertebral artery type(CSA)model rats and explore the mechanism of adrenal gland's regulation of CSA.Methods Ten SPF male Wistar rats were randomly divided into model group and blank group.The CSA model was established by compound modeling method.The total RNA was extracted from the pituitary;the gene expression profile was detected by whole gene chip,ontology(GO),and signal pathway analysis.Results Compared with the normal group,the differential genes'expression profile analysis showed that the total number of the differential genes was 321(fold change︱ ＞ 2,P＜0.05),with 203 up-regulated genes and 118 down-regulated genes.A total of 1 294 genes rich in GO function were involved in the regulation of intercellular signal activity and nerve cell function;the stress response to external stimuli;and the regulation of coagulation function,angiogenesis, endometrial system,and cell cycle;There were 145 signal transducers,including adipocytokine signaling pathway, TGF-β signal transduction,AMPK signaling pathway,PPAR signal pathway,Wnt signaling pathway,and MAPK signal transduction pathway.Conclusion The pituitary regulates CSA mainly through the inflammatory stimulation,immune regulation,regulation of vertebral artery function,and endometrial system.
ABSTRACT
M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.
ABSTRACT
Based on the gene sequence of AIV matrix protein 2(M2) and two cytotoxic T-lymphocyte epitopes derived from nucleoprotein,the prokaryotic expression vector pET-3M2e-NP1-NP2 was constructed.The target gene was expressed in the solvable form in BL21(DE3) when induced with 1.0 mmol/L IPTG and Western blot analysis showed that the expression product had good immunogencity.Purified fusion protein was mixed with various amount of adjuvant,including Freund's,Vash oil and chitosan,and then immunized 20-day-old chicken by intramuscular injection and boosted 3weeks later.Blood samples were collected weekly following the primary vaccination.The anti-M2e antibody was detected with ELISA method with the synthesized peptide as coated antigen;the neutralizing ability of anti-serum was evaluated on MDCK cell line and chick embryo,the CD4+ and CD8+ T lymphocyte amounts in peripheral blood of immunized chicken was measured by flow cytometry.Results showed that the fusion protein can induce immunological reaction,and the antibody can bind with the viral M2 protein that expressed on the surface of MDCK cells.Flow cytometry result showed that CD4+ and CD8+ T lymphocyte in peripheral blood increased obviously following immunization(P