ObjectiveTo observe the prevalence of fundus tessellation in college students with high myopia and analyze the relationship of fundus tessellation and ocular biological parameters.MethodsA cross-sectional study. A total of 202 eyes of 161 individuals were included in the study. Among them, there were 49 males and 112 females with the average age of 19.73 ± 1.12 years old, and the average spherical equivalent of －7.39 ± 1.12 D. All participants underwent computer optometry, non-mydriatic fundus photography, OCT, OCT angiography (OCTA) examination and axial length (AL) measurement. The non-mydriatic fundus camera was used to take the photo of fundus. Fundus tessellation was differentiated to no leopard eye fundus (grade 0), mild leopard eye fundus (grade 1) and middle leopard eye fundus (grade 2) and for severe leopard eye fundus (grade 3). The lenstar was used for eye axis measurement. The choroid, retinal thickness and microvessel density of the macular fovea at the posterior pole of the fundus were measured using a swept-frequency source optical coherence tomography scanner. According to the ETDRS, the choroid within 6 mm of the fovea was divided into 3 concentric circles centered on the macula, which were the central area with a diameter of 1 mm, the inner ring area of 1-3 mm and the outer ring area of 3-6 mm. The outer ring area of 3-6 mm included a total of 9 zones. The inner ring and outer ring 4 regions were superior, inferior, nasal and temporal, respectively. The distribution characteristics of choroid and retinal thickness in different regions and the density of superficial microvessels in the macular area of the retina were observed. Bivariate correlation analysis was used to analyze the relationship of fundus tessellation and ocular biological parameters.ResultsAmong 202 eyes, 37 eyes with leopard pattern fundus with 0 grade (18.32%, 37/202), 165 eyes with grade 1 to 3 (81.68%, 165/202), of which grade 1, 2 and 3 were respectively 125 (61.88%, 125/202), 28 (13.86%, 28/202), 12 (5.94%, 12/202) eyes. The thickness of the retina, both horizontally and vertically, increased first and then decreased from the nasal side to the central area, was lowest in the central area, then increased and then decreased; the overall thickness of the temporal side was slightly lower than that of the nasal side, and the overall thickness of the lower part was slightly lower than the upper part. The choroidal thickness gradually thickened from the nasal side to the temporal side in the horizontal direction; it gradually decreased in the vertical direction from the top to the central area, then increased and then decreased. The SCP blood flow density in the central area in the horizontal and vertical directions was lower than that in other areas. In multivariate regression analysis, Leopard-like fundus classification and AL (β=0.291, OR=1.338, 95%CI 1.064-1.682, P=0.013), SCP blood flow density in the central area of the macula (β=0.080, OR=1.084, 95% CI 1.006-1.167, P=0.034) and choroidal thickness (β=－0.033, OR=0.968, 95%CI 0.960-0.975, P＜0.001) were related.ConclusionsPatients with high myopia have a higher prevalence of tessellation. The deepening of tessellation is related to the thinning of the choroid thickness, the growth of the eye axis, and the larger density of the superficial microvessels in the fovea.
With the advancement of molecular biology technology and the development of genetics, the viral vector system has been continuously improved and optimized. The viral vector system has gradually become one of the best carriers in ophthalmic gene therapy. Adenovirus vector has the characteristics of transient expression and plays an important role in reducing corneal immune response. Lentiviral vector has the characteristics of stable and high efficiency and can be expressed slowly in the body for a long time.Adeno-associated virus vector has the characteristics of low immunogenicity, high efficiency and precision and can infect a variety of retinal cells. The combined use of adeno-associated virus vector and CRISPR-Cas9 provides new methods for precise treatment of ophthalmic genetic diseases. The advent of viral vectors has significantly increased the potential of gene therapy and has unparalleled advantages over traditional therapies. We have reason to believe that virus-based gene transduction technology will soon achieve clinical application in the near future, and a large number of difficult ophthalmic problems will be solved by then.
With the development of life sciences and informatics, bioinformatics is developing as an interdisciplinary subject. Its main application is the relationship between genes and proteins and their expression. With the help of genomics, proteomics, transcriptomics, and metabolomics, researchers introduce bioinformatics research methods into fundus disease research. A series of gratifying research results have been achieved including the screening of genetic susceptibility genes, the screening of diagnostic markers, and the exploration of pathogenesis. Genomics has the characteristics of high efficiency and accuracy. It has been used to detect new mutation sites in retinoblastoma and retinal pigment degeneration research, which helps to further improve the pathogenesis of retinal genetic diseases. Transcriptomics, proteomics, and metabolomics have high throughput characteristics. They are used to analyze changes in the expression profiles of RNA, proteins, and metabolites in intraocular fluid or isolated cells in disease states, which help to screen biomarkers and further elucidate the pathogenesis. With the advancement of technology, bioinformatics will provide new ideas for the study of ocular fundus diseases.
ObjectiveTo observe the expression of S100A8 in plasma exosomes, microvesicles (MV), plasma and vitreous in patients with diabetic retinopathy (DR), and verify it in a diabetic rat model, and to preliminarily explore its role in the occurrence and development of DR.MethodsA case-control study. From September 2018 to December 2019, a total of 73 patients with type 2 diabetes, hospitalized patients undergoing vitrectomy, and healthy physical examinations in the Tianjin Medical University Eye Hospital were included in the study. Among them, plasma were collected from 32 patients and vitreous fluid were collected from 41 patients, which were divided into plasma sample research cohort and vitreous sample research cohort. The subjects were divided into simple diabetes group (DM group), non-proliferative DR group (NPDR group) and proliferative DR group (PDR group) without fundus changes; healthy subjects were regarded as normal control group (NC group). In the study cohort of vitreous samples, the control group was the vitreous humor of patients with epimacular membrane or macular hole. Plasma exosomes and microvesicles (MVs) were separated using ultracentrifugation. Transmission electron microscopy, nanometer particle size analyzer and Western blot (WB) were used to characterize exosomes and MVs. The mass concentration of S100A8 was determined by enzyme-linked immunosorbent assay. Eighteen healthy male Brown Norway rats were divided into normal control group and diabetic group with 9 rats in each group by random number table method. The rats of diabetes group were induced by streptozotocin to establish diabetic model. Five months after modeling, immunohistochemical staining and WB were used to detect the expression of S100A8 in the retina of rats in the normal control group and the diabetes group. t test was used for the comparison of measurement data between the two groups. Single-factor analysis of variance were used for the comparison of multiple groups of measurement data.parison of measurement data between the two groups. Single-factor analysis of variance were used for the comparison of multiple groups of measurement data.ResultsExosomes and MVs with their own characteristics were successfully separated from plasma. The concentrations of plasma exosomes and vitreous S100A8 in the PDR group were higher than those in the NPDR group, DM group, NC group, and the difference was statistically significant (P=0.039, 0.020, 0.002, 0.002, P＜0.000,＜0.000). In the plasma sample cohort study, It was not statistically significant that the overall comparison of the S100A8 mass concentrations of plasma and plasma MV in the four groups of subjects (F=0.283, 0.015; P=0.836, 0.996). Immunohistochemical staining showed that retinal ganglion cells, bipolar cells, cone rod cells and vascular endothelial cells in the diabetic group all expressed S100A8 protein. Compared with the normal control group, the expression level of S100A8 in the retina of the diabetic group increased, and the difference was statistically significant (t=8.028, P=0.001).ConclusionsThe level of S100A8 protein in circulating exosomes increases significantly with the severity of DR in patients with type 2 diabetes. S100A8 may be an influential factor in the inflammatory environment of DR and a potential anti-inflammatory therapeutic target.
ObjectiveTo compare the clinical features of patients with acute Vogt-Koyanagi-Harada syndrome (VKH syndrome) of optic disc swelling (ODS) and serous retinal detachment (RD).MethodsA retrospective clinical study. From January 2013 to November 2019, 212 patients with acute VKH syndrome diagnosed in the Department of Ophthalmology of Shanghai Xuhui District Central Hospital were included in the study. Among them, there were 105 males (210 eyes) and 107 females (214 eyes). The average age was 40.84±13.90 years. All affected eyes were examined by BCVA, FFA, and OCT. The standard logarithmic visual acuity chart was used for BCVA examination, which was converted into logMAR visual acuity in statistics. According to the changes in the fundus, the patients were divided into the ODS group and the RD group, 36 patients with 72 eyes (16.98%) and 176 patients with 352 eyes (83.02%), respectively. The independent sample t test was performed to compare the age of onset, visit time and BCVA of the two groups of patients, the χ2 test was performed to compare the count data.ResultsAmong the 72 eyes of 36 patients in the ODS group, there were 16 males with 32 eyes (44.44%), 20 females with 40 eyes (55.56%). The average age was 40.56±16.57 years, the average visit time was 22.47±19.98 days, the average logMAR BCVA was 0.68±0.53. Among the 352 eyes of 176 patients in the RD group, there were 89 male patients with 178 eyes (50.56%), and 87 female patients with 174 eyes (49.43%). The average age was 40.90±13.34 years, the average visit time was 17.25±24.40 days, the average logMAR BCVA was 0.80±0.56. The average age (t=－0.116), gender composition ratio (χ2=0.448), average visit time (t=1.204), average logMAR BCVA (t=－1.661) comparisons between the two groups showed no statistically significant differences (P＞0.05). There was no statistically significant difference in the average logMAR BCVA between the RD group and the ODS group of different eyes (t=0.227, 0.810; P＞0.05). There were 50 (69.44%, 50/72) and 272 (77.27%, 272/352) eyes in the ODS group and RD group with inflammation of the anterior segment. There were anterior segment reactions between the two groups. There was no statistically significant difference in the number of eyes (χ2=1.003, P＞0.05). There were 34 (19.32%, 34/176) and 2 (5.56%, 2/36) patients with headache and hearing loss, respectively. The comparison of the number of patients with headache and hearing loss between the two groups showed statistically significant differences (χ2=4.015, P＜0.05).ConclusionCompared the patients with ODS acute VKH syndrome, the patients with serous RD acute VKH syndrome are more likely to have extraocular symptoms such as headache and hearing loss.
ObjectiveTo detect the protein expression change in the proliferation of human retinal microvascular endothelial cells (hRMECs) stimulated with 4-Hydroxynonenal (4-HNE).MethodshRMECs were in a logarithmic growth phase, and then were separated into 4-HNE-stimulated group and negative control group. The concentration of 4-HNE included 5, 10, 20 and 50 μmol/L in 4-HNE-stimulated group, while the negative control group was added in the same volume of ethanol (the solvent of 4-HNE). Then the cells were stimulated with 4-HNE for 24 hours following by CCK-8 kits incubating for 4 hours to detect absorbance. It was found that 10 μmol/L 4-HNE had the most obvious effect on the proliferation of hRMECs. Therefore, the cellular proteins from 10 μmol/L 4-HNE-stimulated group and negative control group were acquired and prepared by FASP sample preparation method. Data independent acquisition was used for data acquisition, and the GO analysis and pathway enrichment were performed for analysis of differentially expressed proteins.ResultsCCK-8 kits detection results showed that the A value of the 10 and 20 μmol/L 4-HNE-stimulated groups were significantly higher than negative control group and 5 μmol/L 4-HNE-stimulated group (F=25.42, P＜0.01), while there were no differences between 10 and 20 μmol/L 4-HNE-stimulated groups, and the A value of 50 μmol/L 4-HNE-stimulated groups was lower than negative control. A total of 2710 quantifiable proteins were identified by peoteomics, and 118 proteins were differentially expressed (fold change＞1.5, P＜0.05). Seventy-two proteins were up-regulated after 4-HNE stimulation, whereas 46 proteins were down-regulated. Particularly, the expressions of Heme oxygenase-1, Sulfoxdoxin-1, Heat shock protein A1B, Thioredoxin reductase-1, Glutathione reductase, ATPase and prothrombin were increased when cells were added in 4-HNE, whereas the expressions of apolipoprotein A1 and programmed cell death protein 4 were decreased. These differentially expressed proteins were mainly involved in the biological processes such as oxidative stress, cell detoxification, and ATPase-coupled membrane transport.ConclusionAfter stimulated with 4-HNE, the oxidative stress, cell detoxification, and ATPase-coupled membrane transport protein expression may change in hRMECs in order to regulate oxidative stress and growth situation.
ObjectiveTo observe the changes of retinal microstructure in lamellar macular hole (LMH) after vitrectomy.MethodsA retrospective clinical observational study. Forty patients (41 eyes) with LMH and received vitrectomy in Ophthalmology Department of Peking University People’s Hospital from January 2014 to September 2018 were included in this study. Among them, 14 patients (15 eyes) were males and 26 patients (26 eyes) were females, with an average age of 67.8±8.6 years. There were 37 eyes with a lens and 4 eyes with an IOL. There were 29 eyes with LMH of tractional type, 7 eyes of degenerative type, and 5 eyes of mixed type. All patients underwent BCVA and OCT examinations. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logMAR visual acuity. The average logMAR BCVA was 0.57±0.27; the mean macular retinal thickness (CRT) was 192.3±108.9 μm, the mean macular thickness (MRT) was 427.5±110.2 μm. Among the 29 eyes of tractional type, there were 17 eyes with retinal cavity, 8 eyes with macular retinoschisis, and 3 eyes with incomplete ellipsoid zone. Among the 7 eyes of degenerative type, there were 5 eyes with lamellar hole-associated epiretinal proliferation (LHEP), 5 eyes with retinal cavity, and 5 eyes with incomplete ellipsoid zone. Among the 5 eyes of mixed type, 2 eyes with LHEP, 1 eye with macular epiretinal membrane, and 4 eyes with incomplete ellipsoid zone. The average follow-up time after surgery was 12.8±5.2 months. Among them, 10 eyes were followed up for equal or greater than 24 months. After the surgery, the same equipment and method before the surgery were used for relevant examination. The changes of BCVA, CRT, and MRT before and after surgery were observed. Continuous variables were compared by t test.ResultsAt the last follow-up, the mean logMAR BCVA was 0.37±0.26. Compared with before surgery, the difference was statistically significant (t=5.98, P＜0.01). The mean CRT and MRT were (245.2±90.8) and (347.0±46.7) μm, respectively. Compared with before surgery, the differences were statistically significant (t=-2.49, -5.24; P＜0.05, ＜0.01). CRT and MRT changed greatly within 6 months after surgery, and then tended to be gentle. Among the 3 eyes with incomplete ellipsoid zone of tractional type before surgery, ellipsoid zone recovered in 2 eyes and partially recovered in 1 eye. Among the 17 eyes with retinal cavity and 8 eyes with macular retinoschisis before surgery, there were still 4 eyes with retinal cavity, but all the retinoschisis were disappeared. Among the 5 eyes with retinal cavity of degenerative type before surgery, there were still 2 eyes with retinal cavity and all the eyes with incomplete ellipsoid zone. Among 10 eyes with a follow-up time of equal or greater than 24 months, the macular ganglion cell complex partially atrophied in 6 eyes, and the nerve fiber layer separated in 2 eyes. There was no full-thickness macular hole after surgery.ConclusionFor most LMH patients, vitrectomy can effectively improve the visual acuity and promote the recovery of retinal microstructure.
ObjectiveTo observe the protective effect of polypyrimidine bundle-binding protein-related splicing factor (PSF) over-expression on RPE cell injury induced by advanced glycation end products (AGEs).MethodsThe human RPE cells cultured in vitro were divided into three groups: normal control group (N group), blank control group (N + AGEs group), empty vector control group (Vec + AGEs group), and PSF high expression group (PSF + AGEs). group). RPE cells in N group were routinely cultured; RPE cells in N + AGEs group were only transfected but did not introduce any exogenous genes combined with AGEs induction; Vec +AGEs group and PSF + AGEs group were transfected with pcDNA The empty vector or pcDNA-PSF eukaryotic expression plasmid was introduced into RPE cells and induced by AGEs. Except the N group, the other 3 groups of cells were transfected accordingly, and were stimulated with 150 μg/ml AGEs for 72 h after 24 h. HE staining and Hoechst 33258 staining were used to observe the effect of high PSF expression on the morphological changes of RPE cells; ROS level detection was used to analyze the effect of PSF high expression on the ROS expression of RPE cells induced by AGEs; MTT colorimetric method was used to detect the high PSF expression Effects on the viability of RPE cells; Western blot was used to detect the effects of different time and dose of PSF on the expression of heme oxygenase 1 (HO-1).ResultsHE staining and Hoechst 33258 staining observation showed that the cells in group N were full in shape, the nucleus was round, the cytoplasm was rich, and the staining was uniform; the cells in N + AGEs group and Vec + AGEs group were reduced in size, the eosinophilic staining was enhanced, and the nucleus was densely densely stained. Pyrolysis and even fragmentation; the morphology of cells in the PSF + AGEs group was still full, the cytoplasm staining was more uniform, and the nucleus staining was uniform. The results of MTT colorimetry showed that high expression of PSF can effectively improve the viability of RPE cells, but this effect can be effectively antagonized by ZnPP, and the difference is statistically significant (F=33.26, P＜0.05). DCFH-DA test results showed that compared with the N + AGEs group and Vec + AGEs group, the ROS production in PSF + AGEs group decreased, the difference was statistically significant (F=11.94, P＜0.05). Western blot analysis showed that PSF protein up-regulated HO-1 expression in a time- and dose-dependent manner. The relative expression level of HO-1 at 24, 48, and 72 h after PSF protein was significantly higher than that at 0 h, and the difference was statistically significant (F=164.91, P＜0.05). The relative expression level of HO-1 under the action of 0.1, 0.5, 1.0, 1.5, and 2.0 μg PSF protein was significantly higher than 0.0 μg, and the difference was statistically significant (F=104.82, P＜0.05).ConclusionPSF may inhibit the production of ROS by up-regulating the expression of HO-1, thus protecting the RPE cells induced by AGEs.