To generate an epitope-mutated foot-and-mouth disease virus (FMDV) as a marker vaccine, the infectious clone pAsia 1-FMDV containing the complete genomic cDNA of Asia 1 type FMDV was used as backbone, the residues at positions 27 and 31 in the 3D gene were mutated (H27Y and N31R). The resulting plasmid pAsia 1-FMDV-3DM encoding a mutated epitope was transfected into BHK-21 cells and the recombinant virus rAsia 1-3DM was rescued. The recombinant virus showed similar biological characteristics comparable with the parental virus. In serological neutralization test the antisera against recombine virus have a good reactivity with parental virus. The antisera against the mutant virus were shown to be reactive with the mutated epitope but not the wild-type one. The results indicated that the two virus strains could be distinguished by western blotting using synthetic peptides. This epitope-mutated FMDV strain will be evaluated as a potential marker vaccine against FMDV infections.
Animals ; Cell Line ; DNA, Complementary ; Epitopes ; genetics ; Foot-and-Mouth Disease Virus ; genetics ; Immune Sera ; Neutralization Tests ; Transfection

Objective:To discuss the effects and mechanism of triptolide regulating Caspase1/GSDMD pathway in bone destruction in collagen-induced arthritis rats.Methods:The rats were divided into blank group, model group, triptolide group, and methotrexate group using a random number table method, with 5 rats in each group. Except for the blank group, all other groups were injected with bovine typeⅡ collagen at the tail root to establish an arthritis model. After 7 days of strengthening immunity, the dosage of triptolide A was calculated based on the body surface area in rats, and the triptolide A group was orally administered with triptolide A for 18 μg/kg, 1 d/time; the methotrexate group received intraperitoneal injection of 0.3 mg/kg methotrexate for 3 days per dose, with continuous intervention for 15 days. The arthritis index (AI) score and toe volume changes of the rats were recorded. The ankle joint histological changes were observed with HE staining, and the ankle joint cartilage and bone changes were observed with ferruginine solid green staining. The contents of IL-18 and IL-1β in serum were determined by ELISA. The mRNA levels of GSDMD, Caspase-1, OPG and RANKL in ankle joints were detected by real-time quantitative PCR. The expressions of GSDMD, Caspase-1, OPG and RANKL in ankle tissues were detected by Western blot.Results:After 1 and 2 weeks of administration, compared with the model group, the AI scores and toe volume values of the triptolide group, and methotrexate group decreased ( P<0.01); in the model group, a large number of inflammatory cell infiltration, synovial pannus formation, and blurred defects of the tide line of cartilage and bone staining were observed. The inflammatory infiltration, synovial pannus formation, articular cartilage and bone destruction were improved to varying degrees in each administration group. Compared with model group, serum IL-18 and IL-1β contents in triptolide group and methotrexate group significantly decreased ( P<0.01), mRNA and protein expressions of GSDMD, Caspase-1 and RANKL decreased ( P<0.01), and mRNA and protein expression of OPG increased ( P<0.01). Conclusion:Triptolide can effectively improve joint inflammation and bone destruction in collagen-induced arthritis rats, and its mechanism may be related to down-regulating the expressions of GSDMD, Caspase-1 and RANKL, and up-regulating the expression of OPG.

Objective To evaluate the safety of Renaissance spine robot assisted system in spinal injury.Methods From March 2014 to May 2016,38 patients with spinal disease received spinal surgery assisted by spine robot system.They were 20 males and 18 females,with an average age of 42 years (range,from 12 to 69 years).There were 10 lumbar fractures,8 thoracic fractures and 20 spinal deformities.Pedicle screw implantation was conducted in 30 patients (PS group) and percutaneous vertebroplasty in 8 (PV group).One side was chosen randomly to use Mazor spine robot assisted system (assisted group) and the opposite side the conventional method (non-assisted group).The anteroposterior and lateral X-rays and CT scan of the lumbar and/or thoracic spine were performed in all patients after surgery.The precision of pedicle screws implantation in PS group was evaluated by the Abul-Kasimhierarchy grading system;location of the puncture trajectory,time used for puncture and radiation exposure time in PV group were evaluated.Results 208 pedicle screws were implanted in PS group,including 120 lumbar ones and 88 thoracic ones.For lumbar pedicle screw implantation,the excellent to good rate was 95.0％ (57/60) in the assisted group,significantly higher than that in the non-assisted group (80.0％,48/60) (P ＜ 0.05).For thoracic pedicle screw implantation,the excellent to good rate was 95.5％ (42/44) in the assisted group,significantly higher than that in the non-assisted group (77.3％,34/44) (P ＜ 0.05).There were 24 puncture trajectories in 8 patients in PV group,showing no pedicle penetration or cement leaking in any case.The mean time used for puncture was 5.5 ± 1.4 min in the assisted group,significantly shorter than that in the non-assisted group (17.8 ± 7.5 min) (P ＜ 0.05);the X-ray exposure time was 14.0 ± 4.0 s in the assisted group,significantly shorter than that in the non-assisted group (22.4 ± 6.0 s) (P ＜ 0.05).Conclusions Renaissance spine robot-assisted system deserves more clinical application,because in spinal surgery it can make pedicle screw implantation more precise and safer,and can reduce operation time and X-ray exposure time in percutaneous vertebroplasty.

Integrin alphavbeta3 plays a major role in various signaling pathways, cell apoptosis, and tumor angiogenesis. To examine the functions and roles of alphavbeta3 integrin, a stable CHO-677 cell line expressing the murine alphavbeta3 heterodimer (designated as "CHO-677-malphavbeta3" cells) was established using a highly efficient lentiviral-mediated gene transfer technique. Integrin subunits alphav and beta3 were detected at the gene and protein levels by polymerase chain reaction (PCR) and indirect immunofluorescent assay (IFA), respectively, in the CHO-677-malphavbeta3 cell line at the 20th passage, implying that these genes were successfully introduced into the CHO-677 cells and expressed stably. A plaque-forming assay, 50% tissue culture infective dose (TCID50), real-time quantitative reverse transcription-PCR, and IFA were used to detect the replication levels of Foot-and-mouth disease virus (FMDV) in the CHO-677-malphavbeta3 cell line. After infection with FMDV/O/ZK/93, the cell line showed a significant increase in viral RNA and protein compared with CHO-677 cells. These findings suggest that we successfully established a stable alphavbeta3-receptor-expressing cell line with increased susceptibility to FMDV. This cell line will be very useful for further investigation of alphavbeta3 integrin, and as a cell model for FMDV research.
Animals ; Animals, Suckling ; CHO Cells ; Cloning, Molecular ; Cricetulus ; DNA, Complementary/genetics/metabolism ; Disease Susceptibility/virology ; Foot-and-Mouth Disease/*genetics/virology ; Foot-and-Mouth Disease Virus/*physiology ; Integrin alphaVbeta3/*genetics/metabolism ; Mice