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Purpose: a large number of extraction the plasmid pVAX1-Ag85A-Ag85B pVAX1; preparation tumor emits red fluorescence visualization of bladder cancer in mice transplanted tumor animal models, the use of in vivo red fluorescence imaging and imaging features of the anti-tumor immune therapy to assess Ag85A and Ag85BDNA vaccines results. Basic research and the development and application of DNA vaccines to provide intuitive, reliable theoretical basis for the application of the tumor optical molecular imaging. Methods: First, the plasmid-containing PVAX1-Ag85A, and pVAX1-Ag85B bacterial (E. coli JM109), a large number of culture in a liquid medium, then extract a large number of plasmid extraction kit plasmid Ag85A, Ag85B, and restriction endonuclease digestion, and finally quantitative experiments to the desired concentration (1mg/ml) spare. Containing the mushroom coral red fluorescent protein (Discosomasp. red fluorescent protein, DsRed) gene plasmid vector using liposome transfected into mouse bladder transitional cell carcinoma (BTT) cells, screening and expanding culture, subcutaneous 615 mouse hind limb inner thighs, the establishment of tumor emits red fluorescence murine bladder cancer xenograft model, modeling the success of 24 mice were randomly divided into PVAX1-Ag85A DNA vaccine group (Ag85A group) pVAX1-Ag85B DNA vaccine group (Ag85B) and saline group, each 8 in each group were from right hind cultivation of tumor cells 13, 20 days after Ag85A intramuscular injection in the right hind leg of mice, Ag85B and saline. And then each group were 6,13,20,27 days after planting, the uptake in vivo fluorescence imaging, imaging features observed transplantation tumor growth and fluorescence intensity. Day 28 the mice were sacrificed in each group were isolated liver, lung, kidney, abdominal lymph node tissue, the fresh the organization made 1mm3 small slice, observed under fluorescence microscope. Results: 1. Successful preparation of recombinant plasmid PVAX1-Ag85A and PVAX1-Ag85B DNA vaccine. The extracted plasmid DNA contained less impurities, and can meet experimental needs. Successfully transfected DsRed gene to mouse bladder transitional cell carcinoma (BTT) (BTT-DsRed) cells transfected cell viability, the red fluorescence with cell passage number increased attenuation. Successfully prepared tumor emits red fluorescence visualization of bladder cancer in mice transplanted tumor animal model of tumorigenic rate of 100%. 4 in the right hind limb tumor cell seeding after the first week and the second week, the red fluorescence imaging in vivo results show no significant difference in tumor cell fluorescence intensity, the difference was not statistically significant (P GT; 0.05). The third week, the red fluorescence imaging in vivo results Ag85B DNA vaccine tumor fluorescence intensity was significantly lower than the saline group, the difference was statistically significant (P lt; 0.05); fourth week Ag85A and Ag85B DNA vaccine group tumor fluorescence intensity values ??were significantly lower than the saline group, the difference was statistically significant (P lt; 0.01), but Ag85B and Ag85A DNA vaccines, tumor fluorescence intensity value of the difference was not statistically significant, P gt; 0.05 each mice liver, lung and kidney were not found in tumor cell metastasis, Ag85B DNA vaccine tumor distant lymph node metastasis (25%) was significantly lower than the saline group (87.5%) and Ag85A group (62.5%), the difference statistically significant (P lt; 0.05). Conclusions: 1. Vivo red fluorescence imaging intake of transplanted tumor fluorescence imaging features, dynamic, objective, sensitive, visual assessment of the real anti-tumor effect of the DNA vaccine on mice bladder cancer xenograft. 2. Ag85A, Ag85B DNA vaccine can significantly reduce the fluorescence intensity of the bladder tumor xenografts demonstrated its-tumor immune response. 3 bladder cancer in mice transplanted subcutaneously for lymph node metastasis, Ag85B DNA vaccine has significantly inhibit tumor distant metastasis.
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