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AIM: To investigate the expression of connective tissue growth factor (CTGF) and α-SMA in human lens epithelium cell (HLEC) line B3 after transfection by liposome-coated siRNA targeting CTGF.METHODS: HLECs were transfected with small interfering RNA (siRNA) targeting CTGF,labeled with 5`-fluorescein isothiocyanate (5`-FITC) and coated with lipofectamine.The transfection ratio was evaluated via fluorescence intensity.Cell counting kit-8 (CCK-8) assay was performed to assess cytoviability of both non-transfected and transfected HLECs.Quantitative RT-PCR,cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection.RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine.The transfection ratio reached 95% at 24h.The proliferation of HLECs was inhibited by siRNA after 72h transfection.The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h.This effect was not found in negative control siRNA.CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs,which is a potential therapeutic strategy for posterior capsular opacification.
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AIM: To study the changes of brain - derived neurotrophic factor ( BDNF ) expression in gene modified bone marrow mesenchymal stem cells ( BMSC) . ●METHODS:BMSC were divided into blank control group ( without transfected BMSC ) , negative control group ( empty vector without BDNF gene transfected BMSC) and experimental group ( BDNF gene transfected BMSC) . The expression of BDNF mRNA in BMSC was measured by Realtime PCR, and the expression of BDNF in BMSC was measured by ELlSA. ●RESULTS:The BDNF mRNA expressions of 3, 4, 5, 6, 7 and 8-generation BMSC cells in the experimental group were higher than those in the blank control group and negative control group. The differences were statistically significant (P3: F=491. 788, P ●CONCLUSION:Long-term expression of BDNF in BMSC can be enhanced by genetic engineering.
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Abstract?AIM: To investigate the influence on blood-retina barrier after intense light exposure in rats.?METHODS: The rats were randomly divided into light exposure group and control group. Rats in light exposure group were exposed in white light (10000lux, 12h on-off, continuing 1-14d) .Rats in control group were only exposed in natural light.The eyes of the rats in the two groups were removed when the rats in light exposure group acceptted intense light after 1, 3, 7 and 14d.We observed the change of retinal structure using hematoxylin-eosin ( HE ) staining, and observed the change of retinal ultrastructure using electron microscope.We quantified the change of retinal vascular permeability using laser scanning confocal fluorescence microscope and spectrophotometry after perfusion of Evans-blue, to evaluate the change of blood-retinal barrier.?RESULTS: At 1d after intense light exposure, the retinal ultrastructure of rats changed, such as denaturation of photoreceptor cells and falling of membranous disc outer segment and thinning of the outer nuclear layer thickness, and so on;and the longer the rats exposure to intense light, the more serious change of the retinal ultrastructure were found.At 3d later, photoreceptor cells began apoptosis.At 14d later, the outer nuclear layer became thinner obviously, and the number of cells reduce obviously.At 1d after intense light exposure, EB leaked from the retinal vascular, and at 14d later the leaking of EB was more obvious.?CONCLUSION: The photoreceptor cell of the outer nuclear layer of retina will degenerate and apoptosis, and the outer nuclear layer will be thinner, and the structure and function of blood-retinal barrier will be destroied, if the eyes of rats exposed in intense light.