In recent years, the subthreshold micropulse laser is a kind of laser mode which is characterized by long intermittence. It achieves effective therapeutic effect while minimizes the damage to tissues. At present, it has been used to treat diabetic macular edema. Early studies suggested that the laser selectively acts on retinal pigment epithelial cells to reduce macular edema by regulating the expression of inflammatory biomarkers, growth factors, heat shock proteins and other substances. In recent years, with the development of research, more and more emphasis has been placed on the role of retinal glial cells. Müller cells are also considered as one of the target cells affected by micropulse laser, but there is no evidence of direct or indirect effects of micropulse laser on Müller cells. In the near future, it is expected that we will have more clinical evidence to confirm the target cells of the micropulse laser, which may be further confirmed by in vitro experiments through Müller cells or Müller cells co-cultured with retina pigment epithelium cells, so as to make a more detailed statement on the mechanism of it.
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.
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.
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.
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 build a small-sample ultra-widefield fundus images (UWFI) multi-disease classification artificial intelligence model, and initially explore the ability of artificial intelligence to classify UWFI multi-disease tasks. MethodsA retrospective study. From 2016 to 2021, 1 608 images from 1 123 patients who attended the Eye Center of the Renmin Hospital of Wuhan University and underwent UWFI examination were used for UWFI multi-disease classification artificial intelligence model construction. Among them, 320, 330, 319, 268, and 371 images were used for diabetic retinopathy (DR), retinal vein occlusion (RVO), pathological myopia (PM), retinal detachment (RD), and normal fundus images, respectively. 135 images from 106 patients at the Tianjin Medical University Eye Hospital were used as the external test set. EfficientNet-B7 was selected as the backbone network for classification analysis of the included UWFI images. The performance of the UWFI multi-task classification model was assessed using the receiver operating characteristic curve, area under the curve (AUC), sensitivity, specificity, and accuracy. All data were expressed using numerical values and 95% confidence intervals (CI). The datasets were trained on the network models ResNet50 and ResNet101 and tested on an external test set to compare and observe the performance of EfficientNet with the 2 models mentioned above. ResultsThe overall classification accuracy of the UWFI multi-disease classification artificial intelligence model on the internal and external test sets was 92.57% (95%CI 91.13%-92.92%) and 88.89% (95%CI 88.11%-90.02%), respectively. These were 96.62% and 92.59% for normal fundus, 95.95% and 95.56% for DR, 96.62% and 98.52% for RVO, 98.65% and 97.04% for PM, and 97.30% and 94.07% for RD, respectively. The mean AUC on the internal and external test sets was 0.993 and 0.983, respectively, with 0.994 and 0.939 for normal fundus, 0.999 and 0.995 for DR, 0.985 and 1.000 for RVO, 0.991 and 0.993 for PM and 0.995 and 0.990 for RD, respectively. EfficientNet performed better than the ResNet50 and ResNet101 models on both the internal and external test sets. ConclusionThe preliminary UWFI multi-disease classification artificial intelligence model using small samples constructed in this study is able to achieve a high accuracy rate, and the model may have some value in assisting clinical screening and diagnosis.
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.
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.