ObjectiveTo investigate the influence of Ataxia-telangiectasia mutated (ATM) activation on cellular oxidative stress induced by high glucose in bovine retinal capillary endothelial cells(BRECs). Methods The BRECs were treated by different culture medium with various glucose concentrations (5 mmol/L glucose, 30 mmol/L glucose, 30 mmol/L glucose+10 μmol/L KU55933) as normal glucose group, high glucose group and treatment group respectively.After the cells incubated for 48 hours, the protein expression of ATM, P-ATM, Mitogen-Activated Protein Kinase P38(P38), P-P38, Extracellular signal-regulated kinases(ERKs), P-ERKs was detected by Western blot; cellular ROS level was detected by Reactive Oxygen Species Assay Kit; propidium iodide/Hoechst staining was used for analysis of apoptosis; the expression of vascular endothelial growth factor (VEGF) in the supernatant was determined by Enzyme-Linked Immunosorbent Assay (ELISA); the paracellular permeability between endothelium cells was detected by FITC-dextran. ResultsCompared with the protein level of P-ATM, P-P38 and P-ERKs in high glucose group increased. Especially, P-P38, P-ERKs expressed much more than in high glucose group. The secretion of VEGF in high glucose group was higher than that in the normal glucose group but less than that in treatment group. The same tendency existed in ROS assay, apoptosis assay and paracellular permeability measuring. ConclusionsHigh glucose induced altered activation of ATM which might play a protective role in cellular oxidative stress. Deficiency of ATM might lead to ROS explosion, cell apoptosis and dysfunction of endothelial barrier. The mechanism might be associated with P38, ERKs and VEGF.
PURPOSE: To produce monoclonal antibodies directed against tumor-associated antigens expressed of retinoblastoma-derived tissue culture cell line SO-RBS0. METHODS:Hybridization was performed and the specificity of the antibody was tested by immunofluorescent and immunohistochemical methods. RESULTS:Two hybridomas secreted specific monoclonal antibody against retinoblastoma cells were produced ,and the isotype of the monoclonal antibody was IgG2a CONCLUION:The monoclonal antibodies were specific and highly active against retinoblastoma cells and might be used as immunoconjugate.
PURPOSE:To carry out preliminary study on immunogenicity or'the retina and provide theoretical bassis of transplant rejection of the retina. METHODS:The allogeneic whole retinal or photoreceptor layer from C57BL/6 mice wer transplanted subcutaneously into BALB/C mice for antigen exposure and delayed hypersensitivity (DH) and modified 51Cr-release assay for specific cytotoxic T lymphoeytes (CTL)were emploied. RESULTS:The allogeneic whole retinal transplantation gave rise to DH(Plt;0.05 )and increased function of CTL of which the killing rate was 33.4% in concentration of 1:90 comparing with negative group (4.8% in 1:90,Plt;0.05)and the risen function could be blocked by anti-CD8. CONCLUSION:We deduce that allogeneic whole retina has immunogenicity and should pay attention to transplant rejection postoperatively.but the photoreceptor layer seems to have no immunogenicity and may be no transplant rejection after its transplantation. （Chin J Ocul Fundus Dis,1997,13: 234-236）
Objective To establish an allogenic intraocular melanoma model and observe its pathological features.Methods Thirty-six kunming mice were devided randomly into 3 groups with 12 ones in each, and allogeneic melanoma cells B16F10(C57BL16) were inoculated into the anterior chamber (AC), vitreous cavity (VC) of right eyes and under the skin (subcutaneous, SC) of the back of right feet of each grup respectively. The incidence of tumor occurance, time of breaking through the eyeball and other general pathologic features of the tumor were observed by slip-lamp biomicroscopy and operating microscopy for continuous 32 days, and the results were statistically analyzed. Pathological examination was given for tumors at last.Results The incidence of tumor occurance in both AC (12/1 2 eyes) and VC group (11/11 eyes) was higher than that in SC group (2/12 feet)（χ2=17.143, P＝0 .000；χ2=16.218, P=0.000）. The time of eyeball diabrosis was 11-13 days in AC group and 13-32 days in VC group, and there was significant difference between these two groups (Log Rank=18.22, P=0.000). The intraocular melanomas could grow progressively, but reduced and fell off when they broke through eyeball and grew in or bit for a period. The average diameter of the tumor after 32 days after inoculation was (2.27±1.97) mm in AC group，(3.82±1.85) mm in VC group and (0.94±2.27) mm in SC group. There was significant difference between VC and SC group (t=3.322，P=0.003). In pathohistological examination, tumor tissue necrosis could be observed at the center of the subcutaneous melanomas but not in intraocular melanomas.Conclusions Allogeneic intraocular melanoma model is successfully established which is convenient, repeatable, and helpful to studying the mechanism of genesis and development of this tumor. (Chin J Ocul Fundus Dis，2003，19：333-404)
Objective:To detect collagen I synthesis activity in the vitreous of PVR induced by macrophages in rabbits. Methods:PC Ⅲ (Procollagen Ⅲ ) concentrations were measured by radioim- munoassay in the vitreous samples of 14 rabbit eyes with experimental PVR and 14 control eyes. Results:The mean PC Ⅲ concentration on the 7th day after macrophage injection as 257.58mu;g/L(range,236.04~266.88mu;g/L,n= 4)and significantly increased on the 14th day later. On the 28th day the mean concentration of PC Ⅲ as 912.23mu;g/L (range, 881.36~943.10mu;g/L ;n= 2). There was a significant difference between the 7th and the 14th, 21st of 28th day statistically(P＜0.05). PC Ⅲ was not detected in control eyes. Conclusion:The PC Ⅲ level in the vitreous of rabbit eyes with experimental PVR increased significantly from the 7th to the 28th day after macrophages injection and is well consistent with the time course of scarring and the development of traction retinal detachment in the PVR model. （Chin J Ocul Fundus Dis,1996,12: 43-44）
Objective To investigate the feasibility of differentiation of invitro induced rat bone marrowderived mesenchymal stem cells（rMSCs） into retinal pigment epithelial （RPE） cells.Methods The rMSCs from BrwonNorway (BN) rats were isolated and cultured by adherent screening method. RPE cells lysate made by repeated freezethawing was put into the rMSCs culture system to identify whether the induced cells could express characteristic label cytokeratin（CK）and S-100 simultaneously or not.Results The growth rate of rMSCs induced by RPE cells lysate was slower and protuberant burr surrounded the fusiform cells. The results of immunoblotting and double immunofluorescence showed that partial induced cells expressed CK and S-100 simultaneously. The result of flow cytometry indicated that 14.1% induced cells expressed CK and S-100 simultaneously.Conclusion Induced by RPE cells lysate, rMSCs can differentiate into RPE cells.