动物造模,药效评价,葡萄膜炎 z-bio 2024-08-15 351

　　1. Model making method: F1 generation was obtained by mating genetically modified positive mice, and F2 generation was obtained by mating sisters. The experimental animals include the first genetically modified mice, F1 and F2 generations.

　　Positive animal testing: ① Target gene integration and copy number testing. This mainly includes extracting mouse tail DNA, screening with 2 pairs of primers by PCR, and further identification by spot hybridization and Southern hybridization. The probe is the 615 kb HLA2B2704 gene, which is directly labeled and detected by alkaline phosphatase kit Positive animal expression detection. Trim the tail of a mouse to collect blood, dissolve red blood cells to obtain surrounding lymphocytes. Immunohistochemistry and flow cytometry were used to detect the expression level of the target DNA. The primary antibodies used were HC10 antibody (anti-HLA2B27 heavy chain specific monoclonal antibody) and HLA-ABC class I (anti human HLA class I molecular specific antibody). The SP immunoassay kit method was performed according to the instructions. FITC labeled sheep anti mouse.

　　2. Observation indicators and analysis

　　(1) Eye examination: Posterior corneal deposition can be seen under slit lamp. Under the dissecting microscope, the edema of the optic disc is not obvious, and there is no significant dilation of retinal blood vessels.

　　(2) Pathological examination of the eyes: proliferation of outer corneal epithelial cells and disordered arrangement of columnar epithelium. The arrangement of collagen fibers in the middle layer is irregular, and the number of cells increases. The inner epithelial cells are enlarged, and epithelial cell shedding can be seen in some areas. The aqueous humor examination showed that the total number of cells in the aqueous humor of mice with anterior uveitis increased, and the proportion of inflammatory cells was significantly higher than that of normal mice. Cell classification includes shed epithelial cells, iris pigment cells, lymphocytes, and a small number of monocytes.