| 1.Objective:Corneal neovascularization lead to decreased corneal transparency so that it can induce vision impair even occoecatio. The complex process of corneal neovascularization is related with multiple factors. Though there are many essays to treat corneal new born vessels, there isn't exact method until now. The corneal alkali burn model was wide used in the investigation of corneal neovascularization to detect various kinds of angiogenesis factors and inhibiting factors. We establish rat corneal alkali burn model to approach the inhibiting effect of radiation on corneal new vessels. We wish to provide a new utility method for clinical application and compare its therapeutic effect with the 1%CsA eye drops which has been used as a routine method so as to make it clear whether it is really potent. We witness its relationship with CD34,Bcl-2,BAX,VEGF. We devided the rats into several groups to find the appropriate dose of radiation and make the clinical safety evaluation.Corneal neovascularization can induce corneal graft failure. Rats corneal transplantation is performed in a high-risk situation between SD and WISTAR rats. Just the same as above we detect the relationship between radiation and VEGF. We make the clinical safety evaluation.2.Content:1).90Sr-90Y ophthalmic applicator designment:The applicator used in this experiment is routine in clinical nuclear medicine. It was bought from atomic energy isotope research institute,tipe:SSR9013, absorbed dose rate:0.046Gy/s,it was detected by radioactive nuclide application treatment healthy protecting standard. The applicator undertook manufacture research,dosimetric medicine research,applicator concavity origin appearance contaminated level,applicator temperature and humidity serviceable range,applicator appearance status,applicator radiation homogeneity,radioautography,the radiogical safety calculation of applicator origin surface.2).Rat alkali Burn model:We established rat alkali burn model in accordance with Benelli U's method .The rats were subgrouped to five groups: Group A:large doseβ-ray irradiating group, 10Gy for once. Group B:low doseβ-ray irradiating group, 7Gy divided for 7days once a day. Group C:low doseβ-ray irradiating group, 10Gy divided for 7days once a day. Group D:1%CsA treating group,1%CsA eye drops for 7days three times a day. We selected 6 rats from each group randomly and observed the corneal inflammation,dropsy,opacitas,neovascularization by slit lamp. We calculate the new vessel response area. The datas were statistical analyzed. The experimental results hinted that the new vessels of group B were limited. It is of statistical significance.3).The changement of HE after corneal alkali burn and CD34,Bcl-2,BAX immunohistochemistry outcoms:The corneal samples of group B,C,D,E 3days,5days,7days after alkali burn were dyed by HE and CD34,BCL-2,BAX immunohistochemistry. CD34,BcL-2,BAX immunohistochemistry blades's gray scale values were measured by high definition chromatic color pathologic analytical system, The values were statistical analyzed. HE pathologic blade results:the corneal vessels of group B,C,D were less than group E. Inflammatory infiltration of group E was seriousy. Group E had the most corneal new vessels.CD34 immunohistochemistry:the group B,C,D were dyeing weakly positive,the group E was dyeing masculine. Bcl-2 and BAX immunohistochemistry:Bcl-2 expressed strong masculine in the group E. Bcl-2 expressed masculine in the group D. Bcl-2 expressed weakly positive in the group B,C.BAX expressed opposite to Bcl-2.4).Reverse transcriptase-polymerase chain reaction (RT-PCR),Western blot and immunohistochemical staining were employed to examine the expression of VEGF after alkali burn. VEGF expressed weakly in the new born vascular endothelial cells,inflammatory cells in group A,B,C,VEGF expressed masculine in the corneal epithelium cells,corneal stroma cells,new born vascular endothelial cells,inflammatory cells in group D. VEGF expressed strong masculine in the corneal epithelium cells,corneal stroma cells,new born vascular endothelial cells,inflammatory cells in group E. RT-PCR results: VEGF mRNA was overexpressed in group E,VEGF mRNA expressed lower in group B and C, VEGF mRNA expressed medium in group D. VEGF Western-blot results:the VEGF expressed in this order:C |
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