Optical coherence tomography (OCT), as a high-resolution, non-invasive, in-vivo image method has been widely used in retinal field, especially in the examination of fundus diseases. Nowadays, the modality has been gradually popularized in most of the national basic-level hospitals. However, OCT is only employed as a diagnostic tool in most cases, ophthalmologists lack of awareness of further exploring the information behind the raw data. In the era of fast-developing artificial intelligence, on the basis of standardized information management, a more comprehensive OCT database should be established. Further original image processing, lesion analysis, and artificial intelligence development of OCT images will help improve the understanding level of vitreoretinal diseases among clinicians and assist ophthalmologists to make more appropriate clinical decisions.
Objective To observe the abnormal expression of alpha;A-crystallin protein in neural retina in type 2 diabetic rats via proteomic technique.Methods Twenty-eight male Sprague-Dawley (SD) rats were randomly divided into the normal control and the diabetic experimental groups with 14 rats in each group.A type 2 diabetes rat model (T2DM) was set up in the diabetic experimental group by feeding high fat diet combined with peritoneal injection of low dose streptozotocin (STZ);the successful diabetes model is with the randomlydetected blood glucose of ＞16.7 mmol/L.The rats in the control group underwent peritoneal injection of equivalent sodium citrate solution and were fed with normal diet.All of the animals were sacrificed by decapitation 56 days after the induction of diabetes.The eyes were enucleated and the neural retina layers were carefully peeled off and preserved.The total neural retinal proteins were extracted from the control and diabetic groups, respectively,and then subjected to two dimensional gel electrophoresis (2-DE).Some different proteins spots were identified by peptide mass fingerprinting (PMF) as well as by tandem mass spectrometric (MS/MS) measurements.Western blot and indirect immunofluorescence (IMF) were used to confirmed that alpha;A-crystallin protein expression was upregulated in diabetic retina.Results An average of （3122plusmn;37） spots in normal retinas and（2702plusmn;21）spots in diabetic group were found by 2-DE image analysis software; about 150 spots in 2-DE gel of diabetic retinae exhibited statistically significant variations (t＞2.77,P＜0.05).Compared with normal rats' retinae, diabetic ones presented 68 protein spots of up regulation expression and 82 of downregulation expression in 2DE gel.Furthermore,20 of the 150 protein spots were identified by mass spectrometry.The points of 2369 and 1048 in 2-DE gel, showing high expression in diabetic retinal tissues, were identified as alpha;A-crystallin via PMF.Western blot validated that the expression level of alpha;A-crystallin in diabetic neural retina was much higher than that in the control group. Significantly increased expression of alpha;A-crystallin in nuclear retina in diabetic group was also observed by IMF. Fluorescence was mainly seen in the retinal nuclear layer;alpha;A-crystallin aggregation was detected in the perinuclear region of neurons.Conclusion The expression of alpha;A-crystallin increases in neural retina of early T2DM rats.
Based on the pathogenic mechanisms of age-related macular degeneration (AMD), tremendous preclinical and clinical trials have demonstrated that cell transplantation which aim to replace impaired retinal pigment epithelium (RPE) with healthy RPE cells is a promising approach to treat AMD. So far, choices of cell sources mainly are autologous RPE, iris pigment epithelium, fetal RPE, human embryonic stem cell-derived RPE and human induced pluripotent stem cell-derived RPE, and some of them are undergoing clinical researches. Grafting manners in cell-based therapies are various including RPE sheet or RPE-choroid complex transplantation, RPE cell suspension injection, and RPE sheet transplantation with scaffolds. This review is limited to cell-based therapies for RPE that damaged first in the progress of AMD and focus on recent advances in cell sources, transplantation methods, preclinical and clinical trials, and the obstacles that must be overcome.
Clustered regularly interspersed short palindromic repeats/Cas system is a powerful genome-editing tool for efficient and precise genome engineering both in vitro and in vivo, with the advantages of easy, convenient and low cost. This technology makes it possible to simultaneously mutate multiple genes in a single fertilized egg, thus to study the gene expression, genetic interaction and gene function. Even though this method is still in its immature stage and its stability is inconclusive, making precision models of ocular diseases through genome editing may provide a positive effect to explore gene targeted therapy in genetic eye disease.