Important role of Rho/ROCK2 pathway in the development of Alzheimer′s disease / 国际药学研究杂志

Rho/ROCK pathway is a ubiquitous singling pathway in organisms,and is involved in many biological processes. In the brain of Alzheimer′s patients,the activities of Rho and Rho associated coiled coil forming protein kinase(ROCK)are up-regulat?ed,which is accompanied by the elevation of Aβ42 level,and the abnormal change of the morphology and function of neuronal process?es,suggesting that the occurrence and development of Alzheimer′s disease(AD)is associafed with the overexpression and excessive activation of Rho or ROCK. Rho/ROCK2 pathway is considered a target pathway for the prevention and treatment of AD,and Rho or ROCK2 also becomes an important target for AD drug development. Numerous studies have revealed that suppressing the expression or decreasing the activity of Rho or ROCK2 can reduce Aβ42-induced neurotoxicity,protect neurons,and slow down the occurrence and de?velopment of AD. Therefore,specific inhibition of ROCK2 has an important significance for the repair of central nervous system dam?age and the treatment of AD. This article reviews several effects of Rho/ROCK2 pathway on the development of AD.

Application of glucose-6-phosphate isomerase and auto-antibodies in diagnosis of rheumatoid arthritis / 中华全科医师杂志

Blood samples were collected from 90 patients with confirmed rheumatoid arthritis (RA), 73 patients with other rheumatic diseases and 50 normal controls. Serum glucose-6-phosphate isomerase (GPI) and anti-cyclic citrullinated peptide (CCP) were detected with ELISA method, rheumatoid factor IgM( RF IgM)was measured with turbidimetric immunoassay and antikeratin antibody (AKA) was determined with indirect immunofluorescence assay. The serum GPI levels of RA group[1.73 (0. 43 -4. 40)mg/L]were significantly higher than those of rheumatic disease[0. 14 (0. 10 -0. 18 )mg/L]and normal controls[0. 12 (0. 09 - 0. 15 ) mg/L], ( H = 18. 13, P ＜ 0. 01 ). The sensitivities of GPI, anti-CCP,RF-IgM, AKA, GPI + CCP, GPI + RF-IgM were 80% (72/90), 57% (51/90), 68% (61/90), 29% (26/90) ,92% (83/90) and 96% (86/90), respectively; the specificities were 97% (119/123) ,95% ( 117/123 ) ,77% (95/123) ,95% ( 117/123 ), 93% ( 115/123 ) and 76% ( 93/123 ), respectively. GP1 level was significantly correlated with CCP and RF-IgM ( r = 0. 674 and 0. 533, P ＜ 0. 01 ), but not correlated with AKA ( r = 0. 12, P ＞ 0. 05 ). The results suggest that serum glucose-6-phosphate isomerase level is a valuable indicator for early diagnosis of rheumatoid arthritis.

Protein Components of Lewy Bodies (LBs) Identified from Synthetic Proteasome Inhibitor (PSI)-induced Inclusions in PC12 Cells by MS Analysis / 中国生物化学与分子生物学报

Proteomic analysis is an effective way to identify protein constituent in Lewy bedy-like inclusions (or aggresome) in vitro. Exposure to synthetic proteasome inhibitor (PSI, 10 μmol/L) for 48 hours was used to induce the formation of cytoplasmic proteineous inclusions (termed as PSi-induced inclusions) in PC12 cells.The proteomic approaches of biochemical fractionation, two-dimensional electrophoresis (2-D) and identification via peptide mass fingerprints (PMF) were deployed, and 20 protein components of LBs were identified,i ncluding 2 proteins involved in the production of synaptic neurotransmitter, 6 subunits of the 26 S proteasome,2 cytoskeleton proteins, 2 subunits of mitochondrial complexes, 1 anti-oxidant protein, and 7 chaperone proteins and (or) chaperone-like proteins. The results suggested that these LB protein components might had been recruited in PSI-induced inclusions formed in PC12 cells under the condition of proteasome inhibition.

Eukaryotic Translation Factors Resided in PSI-induced Inclusions in PC12 / 中国生物化学与分子生物学报

Lewy body (LB), an eosinophilic inclusion localized in the neuronal perikaryon, consists of a wide range of proteins, including the consistent organization and the selective composition. Treatment of PC12 cells with synthetic proteasome inhibitor (PSI) at 10 μmol/L for 48 hours induced the formation of inclusions, which were detected by eosin staining and immunostaining for α-synuclein. To investigate the potential new components of PSI-induced inclusions in vitro, pure intact inclusions were successfully obtained by fractionation and subjected to two-dimensional electrophoresis (2-DE) then analyzed with unequivocal matrixassisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Eukaryotic translation initiation factor 3 subunit 5 (eIF-3ε), eukaryotic elongation factor 2 (eEF-2) and mitochondrial elongation factor Tu (EF-Tumt) were identified. The results suggest that 3 eukaryotic translation factors recruited in PSI-induced inclusions may influence formation of the intermediate organelles following the inhibition of proteasomes.

Construction and immunogenicity of recombinant fowlpox vaccines coexpressing HA of AIV H5N1 and chicken IL18.

cDNAs of the HA genes of subtype H5N1 AIV were fused to form a single open reading frame, designated H5HA-H7HA. The H5HA-H7HA cDNA and chicken Interleukin-18 (IL18) were inserted into the fowlpox virus (FPV) expression vector pUTA-16-LacZ to produce pUTAL-H5-H7-IL18. cDNA of H5N1 AIV HA was inserted into the FPV expression vector pUTA2 to create the recombinant expression plasmid pUTA2-H5. Plasmids were then co-transected into CEF cells. The two recombinant fowlpox viruses (rFPV) were produced by three cycles with the BrdU and verified by RT-PCR, IFA and Western blotting. One-day-old specific pathogen free (SPF) chickens and 7-day-old commercial Leghorn egg-laying chickens were inoculated with 10(6) PFU recombinant or parental fowlpox vaccine viruses by wing-web puncture. Hemagglutination inhibition (HI) antibody titer and nonspecific cellular immunity level were assessed after 1-3 weeks post-immunization. We found that all rFPV-vaccinated groups produced HI-specific antibodies, and the level of cellular immunity induced by the rFPV-H5-H7-IL18 strain was significantly higher than that induced by rFPV-H5HA. At 3 weeks post-inoculation, immunized SPF and Leghorn chickens were challenged with H5N1 HP AIV. The rFPV-H5-H7-IL18 vaccine strains were able to induce complete (10/10) protection, while the rFPV-H5HA vaccine strain induced (9/10) protection. Cloacal swabbing samples were collected from immunized leghorn chickens during the first week post-challenge; no shedding was found in the rFPV-H5-H7-IL18 vaccinated group. The rFPV-H5-H7-IL18 vaccinated group displayed significantly increased weight gain relative to the rFPV-H5HA group. This study reports a significant step in the further development of new AIV vaccines.

Construction of recombinant baculovirus vector of HIV-1 gag-gp120 chimeric gene / 吉林大学学报(医学版)

Objective:A new recombinant baculovirus transfer vector was constructed, in whic h a recombinant gene fragment encoding HIV-1 gag-gp120 chimeric gene was inse rted.Methods:After HIV-1 gag gene and gp120 gene were linked,the recombinant baculov irus vector was constructed,and the DNA recombinant technique and the E coli /baculovirus system were used.Results:Gel electrophoresis of DNA analysis showed that the genes were recombine d correctly.Electron microscope showed that the recombinant baculovirus reproduc ed in a great quantity.Conclusion:Recombinant baculovirus vector which HIV-1 gag-gp120 chimeric gene fragment was inserted in was constructed successfully.This vector is useful in study of the expression and the biological function of the HIV-1 Gag-gp120 ch imeric protein.