Objective To investigate the ingestion, metabolism and subcellular localization of indocyanine green (ICG) in human retinal epithelial (R PE) cells.Methods RPE cells were incubated with 0.25 mg/ml ICG under the condition of 37oC in the camera. The ICG granule and ultrastructure of RPE cells were observed under the electron microscopy after 1, 4, and 24hour incubation, and the ICG autofluorescence was detected by fluorescence microscopy after the incubation for 1, 2, 4, 8, 12, 24, and 48 hours, respectively. The ab sorbency (A value) of ICG solution was measured at 805 nm with ultraviol et/v isible specrtrometer. The standard curve of concentration of ICG was drawn and the related equation of concentration of ICG and the A value was calculated. After being incubated for 1, 2, 4, 8, 12, 24, 48, and 72 hours, respectively, the A value of supernatant fluid was calculated according to the equation. Aft er incubated with ICG for 24 hours, one sample was observed under electron microscope and fluorescence microscope per week to evaluate the metabolizable period of ICG .Results ICG granules were distributed evenly after entering the RPE cells. After incubated with 0.25 mg/ml ICG for 24 hours, no significant change of the ultrastructure of the RPE cells was found. ICG granules accu mulated in the cells as the time goes by and reached the peak after 24 hours, and then they decreased because of the slowdown of the metabolism. Few ICG was still remained in the cells 1 week later Conclusions RPE cells may take in ICG actively. ICG metabolizable period in RPE cells is long, which may be one of the mechanisms of the toxicity of ICG to the retina in the vitreous operation.(Chin J Ocul Fundus Dis，2004，20：179-181)
Objective To establish a method for primary culture of iris pigment epithelial cells(IPE). MethodsEnzyme-Assisted microdissection was used to isolate and cultivate the IPE cells.An identification was made with microscopic and immunohistochemical observations.Results IPE were successfully sultured and showed on differences with RPE in primary culture and subculture.ConclusionEnzyme-Assisted microdissection is a reliable and quick method for the isolation of IPE.
Objective To investigate the inhibitive effect of E2F decoy oligodeoxynucleotides (E2F decoy ODNs) on cultured human retinal pigment epithelial (HRPE) cells.Methods E2F decoy ODNs or scramble decoy ODNs at varied concentrations were put into the HRPE cells mediated by lipofectamineTM2000. The proliferative activity of HRPE was detected by methythiazolyl-terazollium assay, and the competitive combinative activity of E2F decoy ODNs and transcription factor E2F was detected by electrophoresis mobility-shift assay. Results The proliferation of HRPE was inhibited markedly by E2F decoy ODNs at the concentration of 0.2 μmol/L (P=0.002) in a dose-dependent manner but not by scrambled decoy. The results of electrophoresis mobility-shift assay showed that the combinative activity of transcription factor E2F was abolished completely by E2F decoy ODNs. Conclusions E2F decoy ODNs may sequence-specifically inhibit the combinative activity of transcripti on factor E2F,and inhibit the proliferation of HRPE cells.(Chin J Ocul Fundus Dis，2004，20：182-185)
Purpose:To evaluate the function of gap junction-mediated intercellular communication in cultured cells of retinal pigment epithelial(RPE) cells from porcine eyes.
Methods:The cultured RPE cells were previously stained by a fluorescent probe 5, 6-carboxy fluorescein diacetate (CFDA) ,and then photobleach the fluorescent molecule in chosed cells. Using laser scanning confocal
microscope (LSCM)to observe fluorescence recovery rate of the RPE cells which located in different condition. The function of gap junction communication was evaluated according to the fluorescence recovery
rate.
Results:The fluorescence recovered after photobleached and the fluorescent density of cells which touching to them descend. The recovery rate per minnte of the cells which the cell number it adjacent to was 1,2 and 3 respectively was 1. 997plusmn;0. 665, 4. 378plusmn;0. 811 and 8. 736plusmn;2. 084. Conclusion:The cultured porcine RPE cells have the function of gap junction communication,and its function proportion is associate to its adjoining cells number.
（Chin J Ocul Fundus Dis,1996,12: 41-42）
Systemic lupus erythematosus is an autoimmune disease involving multiple organs of the body. Lupus nephritis is one of the most serious organ manifestations of systemic lupus erythematosus. Vimentin, a member of the intermediate filament protein family, is involved in the pathogenesis of many autoimmune diseases, including lupus nephritis. More and more studies have shown that vimentin plays an important role in the pathogenesis of lupus nephritis, and has an important influence on the disease development, treatment and prognosis of lupus nephritis. This review focuses on the structure, function and post-translational modification of vimentin, the relationship between vimentin and the pathogenesis of lupus nephritis, and the significance of vimentin expression levels in renal tissues, serum and urine, in order to provide theoretical basis for future mechanism research and clinical application.
Objective To investingate the ultrastructural changes of retinal pigment epithelium(RPE) and its permeability in spontaneously hypertensive rats(SHR)and explore the relation between these changes and hypertensive retinopathy.MethodsThe ultrastructure of RPE cells in the SHR aged five,six,seven months wasobserved with transmission electronmicroscope and compared to its normotensive control strain(WKY) with the same age.Then,lanthanum tracer procedures were carried out to investigate pathological changes of the blood-retinal barrier.Results (1)In SHR the main pathological changes involved swelling of mitochondria,enlargement of endoplasmic reticula,decrease of RPE cell infolding,and sparseness of microvilli.These degenerations were more serious in older rats with higher blood pressure.(2)The breakdown of outer blood-retinal barrier with permeation of lanthanum tracers were evident in SHR aged six or seven month,however,in WKY and five-month SHR the traces were prevented from passing by tight junctions.ConclusionThe degeneration of RPE owing to ischemia and anoxia arises in early periosd of hypertensive retinopathy.The pathological changes of ultrastructure and permeability might interact with the damage of visual cells and play a main role in the hypertensive retinopathy.
Objective To study the construction feasibility of a biodegradable artificial esophagus by the squamous epithelial cells and the myoblast cells seeded on the small intestinal submucosa(SIS) and to investigate the growth patternand angiogenesis of the co-cultured human embryonic squamous epithelial cells and the skeletal myoblasts in vivo. Methods The squamous epithelial cells and the myoblast cells were obtained from the 20-week aborted fetus. Both of their cellswere marked by 5-BrdU in vitro.The isolated cells were then seeded on the SIS and co-cultured in vitro for 24 hours, and then the compound of the cells and the SIS was transplanted into the subcutaneous tissue of the athymismus mice. The observation on the morphology and the cytokeratin AE3 and α-actin specified immunohistochemistry of the squamous epithelial cells and the myoblastcells was performed at each of the following time points: 3 days, 1 week, 2 weeks, and 3 weeks after transplantation. Results The morphological observation indicated that the cultured cells could penetrate into the small intestinal submucosa and form several-layered cell structures, and that the compound of the cells and the SIS could have angiogenesis within 2-3 weeks. The 5-BrdU specified immunohistochemical observation suggested that the cells growing in the small intestinal submucosa scaffold might be the cells transplanted.The cytokeratin AE3 specified and α-actin specified immunohistochemical studies demonstrated that the transplanted cells could differentiate in vivo. Conclusion It is possible to fabricate the framework of a biodegradable artificial esophagus with the epithelial cells and the myoblast cells seeded on the small intestinal submucosa.
