ABSTRACT
In order to construct a recombinant replication deficient human type 5 adenovirus (Ad5) expressing a foot-and-mouth disease virus (FMDV) capsid protein, specific primers for P12A and 3B3C genes of FMDV-OZK93 were synthesized. The P12A and 3B3C genes were then amplified and connected by fusion PCR, and a recombinant shuttle plasmid pDC316-mCMV-EGFP-P12A3B3C expressing the FMDV-OZK93 capsid protein precursor P12A and 3B3C protease were obtained by inserting the P12A3B3C gene into the pDC316-mCMV-EGFP plasmid. The recombinant adenovirus rAdv-P12A3B3C-OZK93 was subsequently packaged, characterized and amplified using AdMaxTM adenovirus packaging system, and the expression was verified by infecting human embryonic kidney cell HEK-293. The humoral and cellular immunity levels of well-expressed and purified recombinant adenovirus immunized mice were evaluated. The results showed that rAdv-P12A3B3C-OZK93 could be stably passaged and the maximum virus titer reached 1×109.1 TCID50/mL. Western blotting and indirect immunofluorescence showed that rAdv-P12A3B3C-OZK93 expressed the FMDV-specific proteins P12A and VP1 in HEK-293 cells. In addition, the PK cell infection experiment confirmed that rAdv-P12A3B3C-OZK93 could infect porcine cells, which is essential for vaccination in pigs. Comparing with the inactivated vaccine group, the recombinant adenovirus could induce higher FMDV-specific IgG antibodies, γ-IFN and IL-10. This indicates that the recombinant adenovirus has good immunity for animal, which is very important for the subsequent development of foot-and-mouth disease vaccine.
Subject(s)
Animals , Humans , Mice , Adenoviridae/genetics , Adenoviruses, Human/genetics , Antibodies, Viral , Capsid/metabolism , Capsid Proteins , Foot-and-Mouth Disease/prevention & control , Foot-and-Mouth Disease Virus/genetics , HEK293 Cells , Recombinant Proteins/genetics , Serogroup , Swine , Viral Proteins , Viral Vaccines/geneticsABSTRACT
Objective@#To evaluate the satisfaction of parents with students eye health services after one year implementation of the "Three-year Action Plan for the Prevention and Control of Myopia in Children and Adolescents in Minhang District (2019-2021)", and to provide a reference for optimizing intervention measures.@*Methods@#A total of 20 176 parents were selected with multi-stage sampling method for the satisfaction evaluate.@*Results@#Health services in appointed medical institutions(6 849, 84.65%), providing feedback on the vision screening(16 915, 83.84%) and procedures in appointed medical institutions(6 704, 82.91%) were the top three terms for parents satisfaction with children and adolescents eye health services, respectively. While the eye behavior and eye environment intervention(8 467, 42.01%), parents informing(7 850, 38.91%) and regular follow-up management(7 222, 33.67%) were the top three that were suggested to improvement. Comparative analysis showed that the school period and the history of previous diagnosis of eye diseases had an impact on the satisfaction of children and adolescents eye health services and the demand for improvement significantly (P<0.05).@*Conclusion@#After one year of intervention, parents are generally satisfied with the eye health services for students. At the same time, students with different characteristics parents focused on difference needs for health services. It urges more refined and personalized eye health services should be provided.
ABSTRACT
Objective To observe the effects of the Diaomo-Zhibeng liquid on the expression of Bcl-2/Bax mRNA and their protein in vitro endometrial glandular epithelial cells in patients of endometrial hyperplasia Methods The endometrial tissue in patients with the pathological diagnosis of endometrial hyperplasia were collected.After isolation,culture and identification,the endometrial cells were divided into four groups:low dosage group (12.5 μg/ml),middle dosage group (25 μg/ml),high dosage group (50μg/ml) and blank control group at randomaccording to the random number table.Three duplicate samples were set for each group.After 48h,the expression of Bcl-2 and Bax were detected by RT-PCR and Western-blot.Results Compared with the blank control group,the expression of Bcl-2 mRNA (1.51 ± 0.07,1.09 ± 0.06,0.93 ± 0.07 vs.1.92 ± 0.08) and protein (0.91 ± 0.02,0.72 ± 0.03,0.62 ± 0.03 vs.1.28 ± 0.02) significantly decreased,while the expression ofBax mRNA (5.73 ± 0.37,8.00 ± 0.23,10.72 ± 0.40 vs.3.27 ± 0.34) and protein (0.45 ± 0.02,0.63 ± 0.02,0.78 ± 0.03 vs.0.32 ± 0.02) sifnificantly promoted in low dosage group,medium dosage group and high dosage group of Diaomo-Zhibeng liquid (P＜0.05).Compared with low dosage group,the expression of Bcl-2 mRNA and protein significantly decreased in middle and high dose group (P＜0.05),but Bax mRNA and protein significantly increased (P＜0.05).Conclusions The Diaomo-Zhibeng liquid can increase the susceptibility of epithelial cells to apoptosis,regulate the balance of cell proliferation and apoptosis tends to apoptosis,have the effect of accelerating cells apoptosis.
ABSTRACT
Objective To explore the neuroprotective effect of dexmedetomidine ( Dex) in rats ex-posed to focal cerebral ischemia/reperfusion ( I/R) induced by middle cerebral artery occlusion ( MCAO) and the possible mechansims.Methods Adult male Sprague-Dawley rats were subjected to MCAO for 90 min followed by reperfusion for 24 h and Dex ( 15μg? kg-1 ) was infused through the left femoral vein im-mediately after the onset of MCAO.The PI3K inhibitor LY294002 ( LY,10 mM,10 μl) or eNOS inhibitor L-NIO ( 1 mg? kg-1 ,10μl) was intracerebroventricular administered before ischemia using a microinjecton. The neurological deficit score,brain edema,cerebral infarct volume,and neuron survivals were evaluated after 24 h of reperfusion.The expression of p-Akt ( Ser473) and p-eNOS ( Ser1177) in the ischemic hemicere-brum was detected by Western Blot.Results Compared with I/R group,the neurological deficit score,cere-bral infarct volume,and the degree of brain edema were significantly reduced in the rats treated with Dex ((2.3±0.4) vs (3.9±0.6),(19.3±3.5)%vs (40.5±5.4)%,(61.8±8.1)%vs (76.3±8.5)%,all P0.05).Conclusion Dex can reduce cerebral injury in rats exposed to focal I/R,which is mediated by the activation of PI3K/Akt-eNOS pathway.