ObjectiveTo observe the value of optical coherence tomography (OCTA) in distinguishing ischemic and non-ischemic branch retinal vein occlusion (BRVO). MethodsA prospective clinical observational study. From January 2020 to January 2021, 44 eyes of 44 patients with BRVO diagnosed in Tianjin Medical University Eye Hospital were included in the study. Among them, there were 24 eyes of 24 males and 20 eyes of 20 females. The macular edema subsided after three consecutive anti-vascular endothelial growth factor (VEGF) drug treatments. All the affected eyes underwent best corrected visual acuity (BCVA), intraocular pressure, ultra-wide-angle fluorescein fundus angiography (UWFFA), and OCTA examination. According to the results of UWFFA, the affected eyes were divided into ischemic group and non-ischemic group, with 22 eyes in 22 patients. The macular area of the affected eye with an OCTA instrument were scaned in the range of 3 mm×3 mm to measure the blood flow density (SVD, DVD), foveal blood flow density (SFVD, DFVD), parafoveal blood flow density (SPFVD, DPFVD), affected hemilateral blood flow density (SHVD, DHVD) and affected quadrant blood flow density (SQVD, DQVD) of the superficial capillary layer (SCP) and deep capillary layer (DCP) of the retina, foveal retinal thickness (CRT), fovea avascular zone (FAZ) area, perimeter of FAZ (PERIM), out-of-roundness index (AI), and blood flow density within 300 μm width of FAZ (FD-300). The two-sample independent t test was used to compare the parameters between the ischemic group and the non-ischemic group. Receiver operating characteristic (ROC) curve analysis was used to measure the area under the curve (AUC) of blood flow density to predict ischemic BRVO, determine the critical value for predicting ischemic BRVO and the corresponding sensitivity and specificity, with AUC＞0.9 as the prediction performance was good. ResultsThe differences of BCVA (t=1.544), intraocular pressure (t=－0.404), SFVD (t=0.444), DFVD (t=－0.812), CRT (t=1.082), FAZ area (t=－0.785), PERIM (t=－0.685), AI (t=1.047) of the eyes in the ischemic group and non-ischemic group were not statistically significant (P＞0.05). The differences of age (t=2.194), SVD (t=－3.796), SPFVD (t=－4.181), SHVD (t=－4.700), SQVD (t=－3.594), DVD (t=－2.324), DPFVD (t=－2.476), DHVD (t=－2.118), DQVD (t=－6.529) and FD-300 (t=－5.116) of the eyes in the ischemic group and non-ischemic group area were statistically significant (P＜0.05). ROC curve analysis results showed that DQVD predicted the AUC of ischemic BRVO the largest (0.917), the best cut-off value was 33.75%, and the sensitivity and specificity were 90.9% and 81.8%, respectively. ConclusionOCTA can quantitatively assess the microvascular structure of SCP and DCP in the macular area of BRVO eyes, and contribute to distinguish ischemic and non-ischemic BRVO.
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 observe the differences of macular microvascular structure between recurrent and non-recurrent macular edema (ME) secondary to central retinal vein occlusion (CRVO) after intravitreal injection of ranibizumab (IVR), and to preliminarily analyze the correlation between recurrence and ME. MethodsA prospective clinical observational study. Forty-five patients (45 eyes) diagnosed as CRVO with ME were included in this study in Tianjin Medical University Eye Hospital from January 2020 to December 2021. There were 22 males (22 eyes) and 23 females (23 eyes). All cases were unilateral. The average age was 61.11±10.88 years old. All patients received IVR treatment once a month for 3 consecutive months. ME were regressive after the initial three treatments. The patients were divided into recurrent group (21 cases, 21 eyes) and non-recurrent group (24 cases, 24 eyes) based on ME recurrence at 6 months after ME resolution. All patients underwent best corrected visual acuity (BCVA), intraocular pressure, and optical coherence tomography angiography (OCTA). OCTA was used to scan the macula in the area of 3 mm×3 mm, and the vessel density (VD) of superficial capillary plexus (SCP), deep capillary plexus (DCP), fovea and parafovea before and after treatment was measured. Foveal retinal thickness, foveal avascular zone (FAZ) area, perimeter of FAZ (PERIM), avascular index of FAZ (AI), VD within 300 μm width of FAZ range (FD-300). Foveal VD included superficial and deep retinal VD (SFVD, DFVD); parafoveal VD included superficial and deep retinal VD (SPFVD, DPFVD). Taking the initial three treatments as the observation time point, the changes of the parameters of the two groups were compared. Comparison between the recurrent and non-recurrent group was performed by two independent sample t-tests. Receiver operating characteristic (ROC) curve analysis was used to measure the area under the curve (AUC) of VD for predicting the recurrence of ME. ResultsThere were no significant differences in age (t=1.350), IOP (t=1.929), SFVD (t=－1.716), DFVD (t=－1.143), CRT (t=－1.207) and AI (t=1.387) between the recurrent and non-recurrent group (P＞0.05). There were significant differences in times of anti-VEGF therapy (t=5.912), BCVA (t=5.003), SVD (t=－4.617), SPFVD (t=－4.110), DVD (t=－5.503), DPFVD (t=－4.772), FAZ area (t=2.172), PERIM (t=2.606) and FD-300 (t=－3.501) between the recurrent and non-recurrent group (P＜0.05). ROC curve analysis showed that the AUC of DVD in predicting the recurrence of ME was highest, with 0.921, and the threshold was 37.65%. The sensitivity and specificity were 91.7% and 85.7%, respectively. ConclusionsThe SVD, SPFVD, DVD, DPFVD and FD-300 in the recurrence group are significantly lower than those in the non-recurrence group, while the FAZ area and PERIM are significantly higher than those in the non-recurrence group. DVD≤37.65% can be used as the best threshold for predicting the recurrence of ME.
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 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.