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@#AIM: To investigate the effects of intravitreal injection of triamcinolone acetonide(TA)on angiogenesis and Notch pathway in photochemistry induced branch retinal vein occlusion(BRVO)model in rats. <p>METHODS: BRVO model rats were induced by photochemistry induction and randomly divided into BRVO model group and TA(1, 7, 21)d groups; at the same time, blank control group was set for comparison. The intraocular pressure of rats was measured by ophthalmotonometer; the condition of rat fundus was observed fluorescein fundus color photography(FFA)and optical coherence tomography(OCT); retinal angiogenesis related factors vascular endothelial growth factor(VEGF)and vascular endothelial growth factor receptor 2(VEGFR2), the protein expressions of Notch pathway important factors Notch 1, Jagged 1 and DLL4 were detected in rat retina by Western blotting(WB). <p>RESULTS: In the normal control group, the fundus vessels were arranged neatly and in a clear state. In the BRVO model group, edema appeared in the fundus, the retina turned white, the arrangement of blood vessels was disordered, the optic disc pit was disappeared, retinal vessels were in the state of vasoconstriction. In TA 1, 7 and 21d groups, edema gradually decreased, blood vessels expansion and bending gradually slowed down, and the optic disc pit was restored. Compared with the blank control group, the intraocular pressure of BRVO model group increased, the thickness of the retina increased at the injured site and 250μm far from injured site, the protein expressions of VEGF, VEGFR2, Notch1 and Jagged1 increased, the protein expression of DLL4 protein was decreased(<i>P</i><0.05). Compared with the BRVO model group, in TA 1d group, the retinal thickness decreased at 250μm far from injured site, the protein expressions of VEGFR2, Notch1 and Jagged1 decreased, the protein expression of DLL4 protein increased; in TA 7d group, the retinal thickness was decreased at the injured site and 250μm far from injured site, the protein expressions of VEGFR2, Notch1 and Jagged1 decreased, the protein expression of DLL4 protein increased; the intraocular pressure of TA 21d group decreased, the thickness of the retina decreased at the injured site and 250μm far from injured site, the protein expressions of VEGF, VEGF R2, Notch1 and Jagged1 decreased, the protein expression of DLL4 protein increased(<i>P</i><0.05). <p>CONCLUSION: Vitreous injection of TA may inhibit angiogenesis by regulating Notch pathway to inhibit the activation of VEGF, thus achieving the retinal protection in BRVO rats.
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@#AIM: To investigate the effects of intravitreal injection of triamcinolone acetonide(TA)on angiogenesis and Notch pathway in photochemistry induced branch retinal vein occlusion(BRVO)model in rats. <p>METHODS: BRVO model rats were induced by photochemistry induction and randomly divided into BRVO model group and TA(1, 7, 21)d groups; at the same time, blank control group was set for comparison. The intraocular pressure of rats was measured by ophthalmotonometer; the condition of rat fundus was observed fluorescein fundus color photography(FFA)and optical coherence tomography(OCT); retinal angiogenesis related factors vascular endothelial growth factor(VEGF)and vascular endothelial growth factor receptor 2(VEGFR2), the protein expressions of Notch pathway important factors Notch 1, Jagged 1 and DLL4 were detected in rat retina by Western blotting(WB). <p>RESULTS: In the normal control group, the fundus vessels were arranged neatly and in a clear state. In the BRVO model group, edema appeared in the fundus, the retina turned white, the arrangement of blood vessels was disordered, the optic disc pit was disappeared, retinal vessels were in the state of vasoconstriction. In TA 1, 7 and 21d groups, edema gradually decreased, blood vessels expansion and bending gradually slowed down, and the optic disc pit was restored. Compared with the blank control group, the intraocular pressure of BRVO model group increased, the thickness of the retina increased at the injured site and 250μm far from injured site, the protein expressions of VEGF, VEGFR2, Notch1 and Jagged1 increased, the protein expression of DLL4 protein was decreased(<i>P</i><0.05). Compared with the BRVO model group, in TA 1d group, the retinal thickness decreased at 250μm far from injured site, the protein expressions of VEGFR2, Notch1 and Jagged1 decreased, the protein expression of DLL4 protein increased; in TA 7d group, the retinal thickness was decreased at the injured site and 250μm far from injured site, the protein expressions of VEGFR2, Notch1 and Jagged1 decreased, the protein expression of DLL4 protein increased; the intraocular pressure of TA 21d group decreased, the thickness of the retina decreased at the injured site and 250μm far from injured site, the protein expressions of VEGF, VEGF R2, Notch1 and Jagged1 decreased, the protein expression of DLL4 protein increased(<i>P</i><0.05). <p>CONCLUSION: Vitreous injection of TA may inhibit angiogenesis by regulating Notch pathway to inhibit the activation of VEGF, thus achieving the retinal protection in BRVO rats.
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AIM:To investigate the natural course and adverse event of branch retinal vein occlusion (BRVO) rat model induced by laser photochemical method. METHODS: Thirty SD (Sprague Dawley) rats were administrated Bangladesh via tail vein. Then 532nm laser (80mW, 100μ m and 100ms) was performed on retinal vein secondary bifurcation of bitamporal optic disk for 50 spots. Electroretinogram (ERG), fundus fluorescein angiography ( FFA), optical coherence tomography (OCT) and fundus (fluorescein) photograph were applied on 1, 3, 5, 7, 10, 14 and 21d after BRVO model constructed. Two rats were sacrificed, respectively, on 1, 5 and 21d after photocoagulation to carry on HE (Hematoxylin - Eosin stain) and VEGF - α (vascular endothelial growth factor - α) immumohistochemical staining. RESULTS: There were three rats died, three rats with severe retinal detachment for excessive bleeding,one rat with retinal sunken, and one rat with cataract. FFA and fundus ( fluorescein) photograph showed that the successful BRVO rat model was 73% (22/30). It was found that the near-end photocoagulation vein became coarse, far - end became diminution on 1d and the photocoagulation vein total recanalization was on 3-7d. ERG showed the amplification of b wave (dark -adaptation 3.0 response) decreased to 0.694士0.042 of control eyes and on 5-7d decreased to rock bottom about 0.487士0.064 of control eyes. Then it increased Aii the time to 0.708士0.0465 of control eyes on 21d. OCT and HE staining found that retinal ganglion cells and outer nuclear layer became edema on 1d in vivo and in vitro.It was observed that the thickness of retina on photocoagulation vein (0μ m or 250μ m) decreased from 5d and there were 3-4 layer cells in ONL on 21d. The expression of VEGF-α at injured site were significantly more than control eyes on 1d and there were no significant difference on 5d;But the expression of VEGF-α were slightly less than control eyes on 21d. CONCLUSION: Photochemical method was a feasible method to establish BRVO rat model. The evolution and development of the BRVO model could partly mimic human BRVO phenomenon. At the same time, it should be improved to increase the successful model rate.
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Objective To investigate the feasibility of immature dendritic cells (imDC)phagocytized psoralen ultraviolet A (PUVA)-treated splenic lymphocytes (PUVA-SP DC)in mice inducing B lymphocytes to be regulatory B cells (Breg)with high secretion of interleukin (IL)-10 (IL-10 +Breg). Methods Bone marrow-derived DC of mice was cultured. Spleen lymphocytes of mice were isolated and treated by PUVA,and turned to be PUVA-SP. The bone marrow-derived imDC was co-cultured with PUVA-SP in vitro to obtain PUVA-SP DC. Splenic B lymphocytes of mice were separated by anti-CD19 magnetic beads and co-cultured with different kinds of DC for 48 hours. The levels of interferon (IFN )-γ,transforming growth factor (TGF)-β,IL-12p70,and IL-10 in the culture supernatant of B lymphocytes,imDC,imDC+B lymphocytes, PUVA-SP DC and PUVA-SP DC +B lymphocytes were measured by enzyme-linked immune absorbent assay (ELISA). The accounts of IL-10 +Breg in B lymphocytes,imDC+B lymphocytes,mDC+B lymphocytes and PUVA-SP DC+B lymphocytes were detected by flow cytometry. Results Compared with the other 4 groups, the level of IL-10 in cell culture supernatant of PUVA-SP DC+B lymphocytes was significantly higher (all in P<0.05). Compared with the other groups,the account of IL-10 +Breg in PUVA-SP DC+B lymphocytes was significantly higher. Conclusions PUVA-SP DC can induce splenic B lymphocytes to differentiate into IL-10 +Breg.
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Objective To design experimental conditions of Riboflavin combining with ultraviolet light illumination for pathogen inactivation and to verify its effect.Methods The effects of multi-factors,such as Riboflavin concentrates,dosage of ultraviolet irradiation,pH value and medium,were observed on E.coli inactivation by photochemical method when it was used as an indicating germ in phosphorus solution(PBS)and taken count by smearing Mac-lonkey agar plates for colony-forming units.Results The reduction of E.coli reached 3.87 logs when the system was treated with 12.5 umol/L riboflavin and 3.0mJ/ml of ultraviolet light at 254nm in PBS medium.Conclusion E.coli in PBS medium was sensitized to low concentrates of Riboflavin and low dosage of ultraviolet irradiation.
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Objective To observe the ability of inactivating human immunodeficiency virus type 1 (HIV-1) in whole blood by visible light combination with methylene blue (MB) at different concentrations. Methods HIV-1 was used as the test virus. The contaminated blood was treated by MB, visible light (640 nm), combined with MB and visible light (640 nm). The test of MT4 cell infection was used to evaluate the virus inactivation efficacy. Results After being treated by MB at the concentrations of 5, 10, and 15 ?mol/L, and then irradiated respectively by visible light (40 000 Lux, 640 nm) for 30, 20, and 10 min, all the added indicated virus at the titer of 10 5.78 TCID 50 of HIV-1 could be inactivated absolutely. Conclusion Methylene blue photochemical method can inactivate HIV-1 in blood effectively.