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 inhibitory effect of lentivirus (LV)-mediated miR-191 on the proliferation and angiogenesis of human retinal vascular endothelial cells (hREC) cultured in vitro.MethodsThe hREC cell lines were cultured in vitro and divided into control group, hypoxia group, LV-empty vector (LV-vector) group, and LV-miR-191 (LV-191) group. The LV-vector group and LV-191 group were transferred to the corresponding lentiviral vector respectively. Flow cytometry was used to detect cell transfection efficiency. Cell Counting Kit-8 (CCK-8) test was used to detect cell proliferation ability. Scarification test and invasion chamber (Transwell) test were used to detect cell migration ability. Matrigel test was used to detect cell lumen formation ability. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the relative expression of miR-191 and relative mRNA expression of its downstream target genes p21, vascular endothelial growth factor (VEGF), cell division protein kinase (CDK) 6, cyclin-D1 (Cyclin D1). Independent sample t test was used for pairwise comparison. ResultsThe results of flow cytometry showed that the transfection efficiency of cells in the control group and the LV-191 group were 0.615% and 99.400%, respectively. The results of CCK-8, scarification, Transwell and Matrigel test showed that, compared with the control group, the number of cell proliferation (t=6.130, 4.606), the cell mobility (t=4.910, 6.702), the number of stained cells on the microporous membrane (t=7.244, 6.724) and the lumen formation ability cells (t=8.345, 9.859) were significantly increased in the hypoxia group and the LV-vector group (P<0.01), while the LV-191 group showed completely opposite performance (t=14.710, 6.245, 5.333, 5.892; P≤0.01). The qPCR test results showed that, compared with the control group and the LV-vector group, the relative expression of miR-191 mRNA in the cells of the LV-191 group was significantly up-regulated (t=44.110, 42.680), the relative expression of Cyclin D1 mRNA (t=29.940, 14.010) and CDK6 mRNA (t=15.200, 7.645) decreased significantly, and the difference were statistically significant (P<0.01); the relative expression of p21 mRNA increased, however, the difference was not statistically significant (t=2.013, 2.755; P>0.05). There was no significant difference in the relative expression of VEGF mRNA in the 4 groups of cells (F=0.966, P>0.05). ConclusionsLV-191 can inhibit the proliferation, migration and tubing of hREC by up-regulating p21 and down-regulating CDK6 and Cyclin D1.
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.
ObjectiveTo screening differentially expressed genes (DEGs) in proliferative diabetic retinopathy (DR) patients to provide new biological therapeutic targets for proliferative DR (PDR) therapy. MethodsA basic research. A total of 3 PDR patients (group PDR) and 3 non-diabetic patients (control group) were enrolled in the study in Tianjin Medical University Eye Hospital in October 2020. In addition, 40 cases of PDR and non-diabetic patients were selected and divided into PDR validation group and control validation group. Peripheral blood validation test was performed in PDR validation group and control validation group; RNA sequencing was performed in PDR group and control group. Transcriptomics (RNAseq) sequencing technology was used to screen DEG in PDR group and control group. The selected DEGs were analyzed by gene ontology (GO) function enrichment analysis, signal pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein-protein interaction network (PPI). The gene expression database was used to find the high-throughput data related to PDR, and multi queue comparison analysis was carried out. The target genes of differentially expressed miRNAs were predicted through targetscan platform, so as to clearly screen the correlation between DEG and PDR. Reverse transcription polymerase chain reaction and Western blot were used to verify the expression of DEG mRNA and protein related to PDR. The relative expression of PDR related DEG mRNA and protein between PDR validation group and control validation group were compared by paired t-test. ResultsA total of 1 337 DEGs were screened by RNAseq sequencing in the peripheral blood of patients with PDR, of which 419 genes were up-regulated and 918 down-regulated. Among them, direct inhibitor of apoptosis protein-binding protein with low isoelectric point (DIABLO), zinc finger and BTB domain containing 10 (ZBTB10), polo-like kinases 3 (PLK3), regulatory subunit 1 (PIK3R1) and B cell translocation gene 3 (BTG3) were differentially expressed in PDR patients. The function of GO was enriched from the analysis of molecular function, biological process and cellular composition. The results showed that DIABLO, ZBTB10, PLK3, PIK3R1, BTG3 were involved in the pathological process related to PDR. KEGG enrichment analysis showed that glucose metabolic pathways such as extracellular matrix receptors, cytokine regulatory pathway, p53 signal pathway and galactose metabolism may be involved in the process of differential genes. The analysis of PPI protein interaction network showed that the larger the DEG-associated protein node, the greater the number of associated nodes. Among them, DIABLO, ZBTB10, PLK3, PIK3R1 and BTG3 played significant roles in the formation of the action network. By comparing and analyzing the existing high-throughput data related to diabetic retinopathy in Gene Expression Omnibus database and predicting by Targetscan platform, it was found that some significant differences in miRNA reported in aqueous humor, vitreous fluid and plasma of DR patients can be regulated by the differential genes found in this study. Compared with the control verification group, the relative expressions of DIABLO, ZBTB10, PLK3, PIK3R1 mRNA and protein in peripheral blood of the PDR verification group were up-regulated, and the relative expression of BTG3 mRNA and protein was down-regulated. ConclusionDIABLO, ZBTB10, PLK3, PIK3R1 and BTG3 are DEGs in patients with PDR, and they can participate in the disease process by regulating the biological processes of cell proliferation, fibrosis and oxidative stress.