Construction and the infection efficiency of BimS lentivirus RNA interference vector / 实用口腔医学杂志

Objective:To construct BimS lentivirus RNA interference(RNAi) vector and to study its infection efficiency by using RNAi technique.Methods:Three interference targets were designed according to the BimS sequence.The single chain primer was annealed into double-stranded oligo sequences,and then connected with vector linearized with Age Ⅰ and EcoR Ⅰ enzyme.The recombinant plasmid was packaged,and the infection efficiency was observed by infecting ACC-2 cells.Results:After amplification,a 337 bp band was appeared in the electrophoresis results of positive clones.Sequence of inserted fragments were identical with the result of DNA sequencing.Restructuring lentivirus was packed in 293T cells,the virus titer was 2 × 108 TU/ml,MOI =20,and the transfection efficiency was 85％.The BmS mRNA relative expression of pFU-GV-BmS-1,pFU-GV-BMS-2 and control group was 0.743 ±0.025,0.466 ±0.023 and 1.266 ±0.042 respectively(between each 2 groups,P ＜0.05).Conclusion:BimS lentivirus virus RNA interference vectors can be constructed,and can efficiently infect ACC-2 cells.

Transgenic tobacco expressed HPV16-L1 and LT-B combined immunization induces strong mucosal and systemic immune responses in mice.

BACKGROUND: Although there are two HPV vaccines have been used to prevent cervical cancer, the cost limits their application in developing countries. The aim of this study was to evaluate the potential value of plant-based HPV16L1 and LTB proteins as a high-efficiency, low-cost and easy-to-use HPV16L1 oral vaccine. RESULTS: Transgenic plant-derived HPV16L1 and LTB were identified, which display potent immunogenicity and biologic activity. Higher levels of specific IgG and IgA levels of HPV16L1 were induced when mice were immunized with L1 combined with LTB by the oral route. The stimulation index (SI) of spleen cells from the L1/LTB-immunized group was significantly higher than that in the L1-immunized group (p < 0.05). The percentage of IFN-γ (+) /IL-4 (+) CD4 (+) T cells from the L1/LTB group was clearly increased compared with that in the L1 and control groups (p < 0.05). METHODS: Plant-expressed HPV16L1 and LTB proteins were extracted from transgenic tobacco leaves, and their biologic characteristics and activity were examined with electron microscopy and GM1-binding assays respectively. Mice were immunized orally with either HPV16L1 or LTB alone or in combination. Induced mucosal and systemic immune responses were detected by ELISA, Hemagglutination inhibition (HAI), lymphocyte proliferation assays and flow cytometry analysis. CONCLUSION: Strong mucosal and systemic immune responses were induced by transgenic tobacco derived HPV16-L1 and LTB combined immunization. This study will lay the foundation for the development of a new type of vaccine to decrease HPV16 infections, which may lead to the prevention of cervical cancer.

DISTRACTION OF OSTEOGENESIS AND LENGTHENING OF CRANIOFACIAL BONE-PRELIMINARY REPORT OF AN EXPERIMENTAL RESEARCH / 药物分析学报

Objective The method of bone lengthening by distraction of callus was used on dog mandible as a model to study the capability and changes of membranous bone during distraction osteogenesis. Methods 15 dogs aged 5～7 months were chosen as the subjects. The operation included a unilateral or bilateral periosteal preserving corti cotomy of mandibular bone, and an introral mini lengthening device were fixed to the buccal side of the mandible. After 7d, the mandible was lengthened lmm/d for 20d and then held in internal fixation for 7～ 140d (1 ～ 20 weeks). Results The dogs were killed at 3 times. Anthropometric measurements, X-ray examination and histological observa tion were conducted and conformed that the distracted bone had formed in the expanded zone successfully. Conclu sion The results suggested that we could use the DO technique in the area of craniofacial clinic. The strong ability of generating new bone in membranous skeleton by DO technique should be further demonstrated in the future.