ObjectiveTo analyze the protein expression changes in the retina of non-arteritic anterior ischemic optic neuropathy (NAION) in rats.MethodsThe rat NAION (rNAION) model was established by Rose Bengal and laser. Twenty Sprague-Dawley rats were randomly divided into 4 groups, the normal control group, the laser control group, the RB injection control group, and the rNAION model group, with 5 rats in each group. The right eye was used as the experimental eye. The retina was dissected at the third day after modeling. Enzyme digestion method was used for sample preparation and data collection was performed in a non-dependent collection mode. The data were quantitatively analyzed by SWATH quantitative mass spectrometry, searching for differential proteins and performing function and pathway analysis.ResultsCompared with the other three control groups, a total of 184 differential proteins were detected in the rNAION group (expression fold greater than 1.5 times and P＜0.05), including 99 up-regulated proteins and 85 down-regulated proteins. The expressions of glial fibrillary acidic protein, guanine nucleotide binding protein 4, laminin 1, 14-3-3γ protein YWHAG were increased. Whereas the expressions of Leucine-rich glioma-inactivated protein 1, secretory carrier-associated membrane protein 5, and Clathrin coat assembly protein AP180 were decreased. The differential proteins are mainly involved in biological processes such as nerve growth, energy metabolism, vesicle-mediated transport, the regulation of synaptic plasticity, apoptosis and inflammation. Pathway enrichment analysis showed that PI3K-Akt signaling pathway and complement and thrombin reaction pathway was related to the disease.ConclusionThe protein expressions of energy metabolism, nerve growth, synaptic vesicle transport and PI3K-Akt signaling pathway can regulate the neuronal regeneration and apoptosis in NAION.
ObjectiveTo explore the multimorbidity of age-related macular degeneration (AMD) and diabetic retinopathy (DR) in diabetic patients, and observe the association between AMD and the two-year progression of DR. MethodsA prospective cohort study. The data were obtained from the Phase I baseline and Phase II follow-up of the Beichen Eye Study, which was conducted from June 2020 to August 2023, and the data from participants with diabetes were extracted for analysis. The baseline study included demographic data, anthropometric indices, ocular biometry, visual acuity, fundus imaging, Lens Opacities Classification System Ⅲ grade, questionnaires and laboratory information, etc., and follow-up was performed after two years. DR diagnosis and grading was performed based on the DR International Classification Criteria, and the eye with the heavier DR classification was taken as the affected eye. According to whether there was new-onset DR or DR progression at the follow-up visit, patients were divided into DR non-progressing group and progressing group. The Wisconsin AMD grading standard was used for AMD diagnosis and grading. Quantitative data were compared using the Mann-Whitney U test, and categorical variables were compared using the χ2 test or Fisher's exact test. Logistic regression models were used to estimate the odds ratio (OR) and 95% confidence interval. Sub-group analysis would be executed if the primary analysis had no significant results. Sensitivity analysis was conducted after the application of multiple imputation for missing data. ResultsA total of 1 190 eligible diabetic patients were included at baseline. The observed prevalence rates were 22.69% (270/1 190) for DR, 25.97% (309/1 190) for AMD, and 6.64% (79/1 190) for DR-AMD co-morbidity. Among the 741 patients who completed the 2-year follow-up, 95 cases (12.82%) were in the DR Progression group and 646 cases (87.18%) were in the non-progression group. Compared with those without AMD, the prevalence of DR In patients with early (24.44%, 66/270), middle (4.07%, 11/270), late atrophic AMD (0.37%, 1/270), and exudative AMD (0.37%, 1/270) showed an increasing trend. However, the differences were not statistically significant (P＞0.05). The results of logistic regression analysis showed that having AMD at baseline was an independent risk factor for DR (OR=1.532, P=0.026). During the follow-up period, subgroup analysis revealed that in AMD patients with an axial length of 22.9-23.5 mm (OR=4.507, P=0.028) OR a platelet-lymphocyte ratio of 99.5-122.0 (OR=4.107, P=0.015), the risk of DR Progression was significantly increased. The results of the sensitivity analysis after multiple imputation of the missing data remained stable. ConclusionAMD in DM patients over 50 years of age is an independent risk factor for DR prevalence and progression.
ObjectiveTo investigate the prevalence and risk factors of tessellation fundus (TF) among Tianjin Medical University students with different refractive statuses. MethodsA cross-sectional study. From September to December 2019, 346 students from Tianjin Medical University were randomly selected and underwent slit-lamp examination, non-cycloplegic auto-refraction, subjective refraction, best-corrected visual acuity, ocular biometric measurement, and non-dilation fundus photography. The differences in the prevalence of TF in basic characteristics and ocular biometric parameters were compared. Based on the equivalent spherical (SE), refractive status was divided into the non-myopia group (SE＞－0.50 D) and the myopia group (SE≤－0.50 D). The myopia group was further divided into mild myopia group (－3.00 D＜SE≤－0.50 D), moderate myopia group (－6.00 D＜SE≤－3.00 D), and high myopia group (SE≤－6.00 D). According to the axis length (AL), the subjects were divided into AL＜24 mm group, 24-26 mm group, and ＞26 mm group. The logistic regression was used to analyze the risk factors affecting TF. Trend tests were performed for each risk factor and TF. ResultsOf the 346 subjects, 324 (93.6%, 324/346) were myopia, of whom 73 (21.1%, 73/346), 167 (48.3%, 167/346), and 84 (24.3%, 84/346) were mild myopia, moderate myopia, and high myopia, respectively; 22 (6.4%, 22/346) were non-myopia. There were 294 (85.0%, 294/346) students with TF in the macula, including 9 (40.91%, 9/22), 58 (79.45%, 58/73), 145 (86.83%, 145/167), and 82 (97.62%, 82/84) in non-myopia, low myopia, moderate myopia, and high myopia group, respectively; 52 (15.0%, 52/346) students were without TF in the macula. There were statistically significant gender differences (χ2=4.47), SE (t=6.29), AL (t=－8.29), anterior chamber depth (Z=－2.62), lens thickness (Z=－2.23), and average corneal radius (Z=－3.58) between students with and without TF in the macula (P＜0.05). Spherical equivalent and axial length were independent risk factors for TF and its severity (P≤0.001). With an increasing degree of myopia, and increasing axial length, the risk of TF increased (P for trend＜0.001). ConclusionsThe prevalence of TF is 85.0% among Tianjin Medical University students. TF is detected in the fundus of no myopia, mild myopia, moderate myopia and high myopia. The degree of myopia is higher, the AL is longer, the possibility of TF is higher.
Objective To observe the effect of different macular edema on the area of foveal avascular zone (FAZ) and its correlation in eyes with branch retinal vein occlusion (BRVO). Methods A total of 72 patients (75 eyes) diagnosed with BRVO were included in the study. There were 40 patients males (42 eyes) and 32 females (33 eyes), with the mean age of (56.00±9.96) years. All the eyes were examined by BCVA, intraocular pressure, slit lamp microscope combined with preset lens, fundus color photography and optical coherence tomography angiography (OCTA). BRVO patients were divided into two groups according to the degree of macular edema: group M300 that was CRT ≥300 μm (38 patients, 39 eyes) and group L300 that was CRT＜300 μm (34 patients, 36 eyes). The macular angiography scan protocol covered a 3 mm×3 mm area. The parameters of macular were measured by the built-in measurement software of the system: (1) area of FAZ, perimeter of FAZ (PERIM), avascular index of FAZ (AI), vascular density within a width of 300 μm around the FAZ region (FD-300); (2) central retinal thickness (CRT); (3) vascular density (VD): the superficial central fovea vascular density (SFVD), the deep central fovea vascular density (DFVD), the superficial hemi-macular vascular density (SHVD), the deep hemi-macular vascular density (DHVD). Spearman test was used to test the correlation between FAZ area and other parameters in each group. Results The FAZ area in group M300 and L300 were 0.388±0.166 mm2 and 0.596±0.512 mm2, respectively. The results of Spearman test showed that the FAZ area of group M300 was positively correlated with PERIM and AI (r=0.932, 0.591; P=0.000, 0.000), negatively correlated with SFVD, DFVD and SHVD (r=−0.490, −0.429, −0.339; P=0.002, 0.006, 0.035). But there was no significant negative correlation between FAZ area and FD-300, CRT, DHVD in group M300 (r=−0.129, −0.053, −0.400; P=0.435, 0.749, 0.395). The FAZ area in group L300 was positively correlated with PERIM and AI (r=0.887, 0.633; P=0.000, 0.000), negatively correlated with SFVD, DFVD, SHVD and DHVD (r=−0.413, −0.643, −0.630, −0.370, −0.411; P=0.012, 0.000, 0.000, 0.026, 0.013). But there was no significant positive correlation between FAZ area and FD-300 in group L300 (r=0.093, P=0.590). Conclusion FAZ area varies with the degree of macular edema. The degree of macular edema is higher, the FAZ area is smaller. FAZ area is positively correlated with PERIM and AI significantly, and negatively correlated with SFVD, DFVD and SHVD.
Objective To observe the difference of macular microvascular features in superficial and deep vascular plexi in patients with branch retinal vein occlusion (BRVO). Methods A total of 63 BRVO patients (63 eyes) were enrolled in this study. There were 28 males (28 eyes) and 35 females (35 eyes). The patients aged from 39 to 74 years, with the mean age of (59.76±8.48) years. All eyes were evaluated by optical coherence tomography angiography (OCTA). The macular angiography scan protocol covered a 3 mm×3 mm area. The focus of angiography analysis included superficial vascular plexus and deep vascular plexus. The following vascular morphological parameters were assessed in these two plexi: foveal avascular zone (FAZ) enlargement, capillary non-perfusion (CNP) occurrence, microvascular abnormalities (MA) appearance, and vascular congestion (VC) signs. The FAZ area was measured by the built-in software. The macular microvascular morphology changes in superficial and deep vascular plexi were compared through McNemar test. Results The superficial and deep plexi showed FAZ enlargement in 43 eyes (68.3%) and 50 eyes (79.4%), CNP in 51 eyes (81%) and 50 eyes (79.4%), MA in 62 eyes (98.4%) and 62 eyes (98.4%), VC in 23 eyes (36.5%) and 52 eyes (82.5%), respectively. FAZ area was (0.55±0.37) mm2. There was no difference in CNP (P=1.000) and MA (P=1.000) between superficial and deep plexi. But, there was difference in FAZ enlargement (P=0.039) and VC signs (P＜0.001) between superficial and deep plexi. Conclusion Deep vascular plexus showed more FAZ enlargement and VC sign than superficial plexus in BRVO patients.
ObjectiveTo observe the relationship between the response to anti-vascular endothelial growth factor (VEGF) drug treatment and single nucleotide polymorphism (SNP) genotype in patients with wet age-related macular degeneration (wAMD). MethodsA retrospective clinical study. From August 2019 to September 2020, 103 eyes of 103 wAMD patients diagnosed in Tianjin Medical University Eye Hospital were included in the study. Among them, there were 59 males (57.28%, 59/103) and 44 females (42.72%, 44/103); the average age was 68.74±7.74 years. The standard logarithmic visual acuity chart was used to detect the Best Corrected Visual Acuity of the affected eye and converted to the logarithmic minimum angle of resolution (logMAR) visual acuity during statistics. Optical coherence tomography was used to detect the central retinal thickness (CRT) of the affected eye. At the same time, the patient's high-density lipoprotein cholesterol (HDL-C) was tested. All eyes were treated with intravitreal injection of anti-VEGF drugs once a month for 3 months. Before the initial treatment, peripheral venous blood from the patient were collected. Interleukin-8 (IL-8), complement C3 gene (C3), complement factor H (CFH), liver lipase (LIPC), cholesterol ester transfer protein (CETP), ATP binding cassette subfamily a member 1 (ABCA1), lipoprotein lipase (LPL), fatty acid desaturation gene cluster (FADS1) SNP. According to gene frequency, genotypes are divided into wild type and mutant type were detected. Qualitative data such as the frequency difference of the genotype distribution in the clinical phenotype and the Hardy-Weinberg equilibrium of the genotype distribution were compared with the Chi-square test or Fisher's exact test. ResultsThere were fewer CRT responders in IL-8 rs4073 mutant (TA+AA) patients than wild-type (TT) [odds ratio (OR)=0.310, 95% confidence interval (CI) 0.106-0.910, P＜0.05). Among them, after the drug stratification test, the proportion of patients with IL-8 rs4073 locus TT genotype in the conbercept treatment group was less CRT non-responders (OR=0.179, 95% CI=0.034-0.960, P=0.033). Patients with LIPC rs2043085 mutant (CT+TT) with BCVA increased ≥0.2 logMAR are more likely than wild-type (CC) (OR=3.031, 95% CI 1.036-8.867, P＜0.05); HDL-C level was significantly lower Compared with wild type (CC), the difference was statistically significant (t=2.448, P=0.016). There was no significant difference in logMAR BCVA and CRT between IL-8 rs4073, LIPC rs2043085 mutant and wild-type patients before treatment (IL-8 rs4073: Z=－0.198, －1.651; P=0.843, 0.099; LIPC rs2043085: Z=－0.532, －0.152; P=0.595, 0.879). C3 rs 225066, CFH rs800292, CETP rs708272, ABCA1 rs1883025, FADS1 rs174547, LPL rs12678919 have no correlation with anti-VEGF drug treatment response. Conclusions Patients with wAMD are treated with anti-VEGF drugs. Those with IL-8 rs4073 locus A genotype may be less responsive to CRT. LIPC rs2043085 locus T genotypes may be relatively more responsive to BCVA.
ObjectiveTo observe clinical phenotypes and analyze the pathogenic genes of Leber congenital amaurosis (LCA). MethodsA retrospective clinical study. From 2019 to 2020, 2 patients diagnosed with LCA by genetic testing in Tianjin Medical University Eye Hospital and their 6 unaffected family members were enrolled in the study. Two patients were from 2 unrelated families, both were probands. The patient's medical history was inquired in detail, slit lamp microscopy, ultra-widefield fundus photography, autofluorescence, and flash visual evoked potential (F-VEP) were performed. Peripheral vein blood (3-5 ml) was collected and genomic DNA was extracted from all study subjects. A total of 381 pathogetic genes associated with inherited retinal diseases, were selected by targeted exome sequencing capture strategy. Sanger sequencing was used to verify suspected pathogenic mutations. Candidate pathogenic mutations were identified after bioinformatics analysis. Sanger sequencing, real-time quantitative polymerase chain reaction and family co-identification were used to confirm the final mutations. ResultsTwo patients were male, aged 3 and 27 years. One case had vision loss in both eyes, accompanied by nystagmus and acupressure eye sign since childhood. The clinical hallmark of the proband (F1-Ⅱ-3) in F1 includes clearly boundary of optic disc, normal retinal blood vessels and macular fovea. The implied period of the maximum forward wave in both eyes of F-VEP was roughly normal, and its amplitude decreased significantly. The phenotype of the proband (F2-Ⅱ-1) in F2 includes optic nerve head pallor, bone-spicule intraretinal pigmentation, “gold-foil maculopathy”, retina patchy hypo-autofluorescence in both eyes. There was no abnormal phenotype in the eyes of the family members. According to the genetic diagnosis, the proband (F1-Ⅱ-3) carried the GUCY2D gene c.835G>A (p.D279N) (M1) and exon 9-19 deletion (M2) compound heterozygous mutations, in which M1 was derived from healthy mother and M2 was derived from healthy father. The proband (F2-Ⅱ-1) carried CRB1 gene c.1576C>T(R526X) (M3) and c.1522T>C (C508R) (M4) compound heterozygous mutations, in which M3 from the healthy father, M4 from the healthy mother. M2 and M4 were novel mutations. ConclusionGUCY2D gene mutations lead to LCA1 type in the F1 family, CRB1 gene mutations lead to LCA8 type in the F2 family; there are significant different phenotypes caused by different pathogenic genes.
Objective To observe the association between the red cell distribution width (RDW)/albumin (ALB) ratio (RAR) and the progression of diabetic retinopathy (DR). MethodsA cohort study. From June 2020 to February 2022, 835 diabetic patients who participated in the Phase II Beichen Eye Study, conducted at the Tianjin Medical University Eye Hospital were included. All participants underwent a two-year follow-up. Data were collected from patients at both baseline and the two-year follow-up, including mydriatic color fundus photography and laboratory tests for RDW and ALB. The RAR was calculated based on these measurements. DR was diagnosed and graded according to the DR International Clinical Severity Scale. Based on the progression of DR, patients were categorized into a non-progression group (689 cases, 83%) and a progression group (146 cases, 17%). Univariate models, as well as models Ⅰ, Ⅱ, and Ⅲ, were constructed after adjusting for various variables. The associations between RAR and its tertiles with the progression of DR were analyzed utilizing multivariate logistic regression analysis. Additionally, subgroup and interaction analyses were conducted to further investigate the relationship. ResultsThere was no significant difference in RDW and ALB levels between patients in the non-progression group and those in the progression group (t=-1.399, 1.954; P > 0.05). However, a significant difference was observed in RAR (t=-2.147, P=0.033). Results from the multivariate logistic regression analysis indicated that, in the fully adjusted model Ⅲ, RAR was an independent risk factor for the progression of DR. Specifically, each unit increase in RAR was associated with a 2.33-fold higher risk of DR progression [odds ratio (OR)= 2.33, 95% confidence interval (CI) 1.20-4.54, P = 0.013]. Compared to the univariate model, the predictive power of the fully adjusted model III for DR progression was 71.3% (area under the curve= 0.713, P < 0.001). Interaction analysis revealed a statistically significant difference in the effect of insulin use on the association between RAR and DR progression (insulin users: OR= 5.83, 95% CI 2.15-15.78, P=0.013). ConclusionsIncreased RAR is associated with a heightened risk of DR progression, and insulin use may influence the relationship between the two.
ObjectiveTo observe the effect of interleukin-8 (IL-8) on the adhesion and migration of retinal vascular endothelial cells (RCEC). MethodsA cell experiment. Human RCEC (hRCEC) was divided into normal control group (N group), advanced glycation end product (AGE) treatment group (AGE group), and AGE-induced combined IL-8 antagonist SB225002 treatment group (AGE+SB group). The effect of AGE on IL-8 expression in hRCEC was observed by Western blot. The effect of SB225002 on hRCEC migration was observed by cell scratch assay. The effects of SB225002 on leukocyte adhesion and reactive oxygen species (ROS) on hRCEC were detected by flow cytometry. Student-t test was performed between the two groups. One-way analysis of variance was performed among the three groups. ResultsCompared with group N, the expression level of IL-8 in cells of AGE group was significantly increased, with statistical significance (t=25.661, P＜0.001). Compared with N group and AGE+SB group, cell mobility in AGE group was significantly increased (F=29.776), leukocyte adhesion number was significantly increased (F=38.159, 38.556), ROS expression level was significantly increased (F=22.336), and the differences were statistically significant (P＜0.05). ConclusionIL-8 antagonist SB225002 may down-regulate hRCEC adhesion and migration by inhibiting ROS expression.
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.