The update of the cognition of fundus diseases is inseparable from the rapid development of fundus multimodal imaging. Especially in recent years, the application of wide and ultra-wide fundus photography, ultra-wide fundus fluorescein angiography, indocyanine green angiography, fundus autofluorescence and optical coherence tomography angiography contribute to observe the peripheral retinopathy more directly. The application of adaptive optics and fluorescence lifetime imaging ophthalmoscopy contribute to have a further understanding of fundus diseases at the cellular and metabolic level. Multimodal imageing reflect the pathological characteristics of the diseases from different angles and levels. At the same time, the digitization and intelligence of fundus images are also developing rapidly. However, there are some problems that the ophthalmologists needs to consider further, such as the correctly understanding the use of multimodal imaging, the application of artificial intelligence, and how to sum up from the images.
ObjectiveTo analyze the early changes of gene expression levels and signaling pathways in 661W cell line under hypoxic conditions and to find potential functional target genes.MethodsThe cultured mouse 661W cells were divided into hypoxia treatment group and normoxia control group. Cells in the hypoxia treatment group were cultured in a three-gas incubator with volume fraction of 1% and 5% CO2 at 37 ℃. Cells in the normoxia control group were cultured in an incubator at 37 ℃ with volume fraction of 5% CO2. High-throughput sequencing technology was used to sequence the transcriptome of 661W cell treated with hypoxia and normoxia for 4 hours to screen for differentially expressed genes (DEG). Clustering heat map analysis, gene ontology (GO) functional enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and protein-protein interaction network (PPI) analysis were performed. The reverse transcription-polymerase chain reaction (RT-PCR) was used to verify the accuracy of the sequencing results.ResultsA total of 506 differentially expressed genes were screened, including 459 up-regulated genes and 47 down-regulated genes. GO functional enrichment analysis showed that the main biological processes of DEG were the cell's response to hypoxia, glycolysis, negative regulation of cell proliferation and apoptosis. hypoxia inducible factor (HIF)-1α pathway, glycolysis, Forkhead box O (FoxO) pathway, Insulin signaling pathway and Adenosine 5'-monophosphate-activated protein kinase (AMPK) pathway were involved in the above process. PPI analysis results showed that hub genes related to hypoxia were Aldoa, Aldoc, Gpi1, Hk2, Hk1, Pfkl, Pfkp, Vhl, Fbxo10 and Fbxo27. The RT-PCR results showed that the relative expression levels of 15 DEG mRNA in the hypoxic treatment group were higher than that of the normoxic control group, and the difference was statistically significant (P＜0.05). The mRNA expression levels of N-myc downstream-regulated gene-1 (Ndrg1), Mt1, and vascular endothelial growth factor A (VEGFA) were time-dependent on hypoxia.ConclusionsUnder hypoxia, DEG is mainly related to glucose metabolism, cell response to hypoxia, regulation of proliferation and apoptosis. HIF-1α pathway, glycolysis, FoxO pathway and AMPK pathway are involved in the early changes of 661W cells under hypoxia. Aldoa, Aldoc, Gpi1, Hk2, Hk1, Pfkl, Pfkp, Vhl, Fbxo10, Fbxo27 may play key roles in the response of 661W cells to hypoxia. Ndrg1, Mt1 and VEGFA could be potential functional target genes for the study of ischemia and hypoxia-related fundus diseases.
Objective To observe the efficacy of lamellar hole-associated epiretinal proliferation (LHEP) flap insertion and autologous blood for degenerative type lamellar macular hole (LMH). Methods Retrospective case review. Twenty-eight eyes of 28 patients with LMH were enrolled in this study. There were 2 males (2 eyes) and 26 females (26 eyes). Best corrected visual acuity (BCVA), medical optometry, slit-lamp biomicroscop, indirect ophthalmoscope, spectral domain optical coherence tomography, b-scan ultrasonography and axial length detection were performed on all patients. Logarithm of the minimum angle of resolution (logMAR) was used to calculate visual acuity. There were 10 eyes (35.7%) with degenerative type LMH (flap insertion group) and LHEP. There were 18 eyes (64.3%) with tractional type LMH (general group). The differences of BCVA, AL, horizontal hole diameter from retina and lens state between two groups were not significant (P＞0.05). The differences of horizontal hole diameter of internal limiting membrane (ILM), central foveal thickness (CFT) and integrity of ellipsoidal zone between two groups were significant (P＜0.05). LHEP flap insertion and autologous blood without ILM peeling were used in eyes of flap insertion group. Vitrectomy combined ILM peeling were used in eyes of general group. The follow-up was ranged from 3 to 14 months. The changes of CFT, central foveal form and logMAR BCVA were observed. Results At latest follow-up, the BCVA of flap insertion group and general group were 0.34±0.27, 0.31±0.29; which significantly better than the preoperative BCVA (Z=−3.519, −4.945; P＜0.001). The CFT of flap insertion group and general group were (200.10±58.78), (226.61±70.49) μm. There was no difference between pre- and post-operative CFT in eyes of general group (Z=−1.455, P=0.146). There was significant difference between pre- and post-operative CFT in eyes of flap insertion group (Z=−2.798, P=0.005). In flap insertion group, regular recovery of the foveal contour occurred in 9 eyes (90.0%), improvement in 1 eyes (10.0%). In general group, regular recovery of the foveal contour occurred in 10 eyes (55.6%), improvement in 8 eyes (44.4%). The closure rate of LMH were 100% both in two groups. Conclusion LHEP flap insertion and autologous blood is an effective treatment of degenerative type LMH.
ObjectiveTo compare the consistency and difference of non-mydriatic two-field 45° ultra-wide field Optos and Clarus500 fundus imaging in a large-scale diabetic retinopathy (DR) screening. MethodsA diagnostic methodology study. From November 2020 to August 2021, 526 eyes of 277 patients with type 2 diabetes who diagnosed in Department of Ophthalmology, Henan Provincial People's Hospital were included in the study. Among them, there were 175 males with 328 eyes and 102 females with 198 eyes; the age was 53±10 years old. The same experienced technician performed the non-mydriatic dual-field 45° fundus imaging and the non-mydriatic ultra-wide-angle imaging system Optos, Clarus500 single-field fundus imaging examination on the patient on the same day, and obtained the dual-field 45° fundus image and Optos, Clarus500 single-field fundus image. The Optos and Clarus500 single-field fundus images in the same area as the dual-field 45° fundus image were captured by Photoshop software, and the Optos and Clarus500 dual-field fundus images were obtained. Subsequently, two experienced ophthalmologists performed interpretation and DR grading of the 5 groups of images, respectively. Images with inconsistent grading results were interpreted by a third ophthalmologist and used as the final grading result. In order to avoid the mydriatic dual-field 45° imaging interpretation results as the standard, the consistency and detection rate difference of the two ultra-wide-angle imaging systems in the rapid DR screening results were evaluated. The weighted Kappa (κ) test was used to analyze the consistency of DR diagnosis between dual-field 45° fundus imaging and Optos and Clarus500 fundus imaging; χ2 test was used to compare the detection rates of DR between different imaging systems. ResultsCompared with the dual-field 45° fundus image, the Clarus500 single-field had a higher DR detection rate (χ2=24.965, P＜0.001), and the Optos dual-field fundus image had a lower DR detection rate (χ2=49.559, P＜0.001). Compared with the DR detection rate of dual-field 45° fundus image, Optos single-field fundus image, Clarus500 double-field fundus image had no significant difference (χ2=2.572, 0.649; P=0.109, 0.421). Compared with Optos, Clarus500 single-field and dual-field fundus images DR detection rate, the difference was statistically significant (χ2=43.214, 61.216; P＜0.001). Consistency assessment of DR grading results: dual-field 45° fundus images and Clarus500 dual-field fundus images (κ value=0.932, 95% confidence interval (CI) 0.907-0.956) were highly consistent; dual-field 45° fundus images and Optos single-field fundus images [κ value=0.474, 95%CI 0.417-0.532], Optos dual-field fundus image (κ value=0.495, 95%CI 0.438-0.551), Optos dual-field fundus image (κ value=0.495, 95%CI 0.438-0.551) and Clarus500 dual-field fundus image (κ value=0.452, 95%CI 0.395-0.506) were moderately consistent; dual-field 45°fundus images and Clarus500 single-field fundus images (κ value=0.354, 95%CI 0.303-0.403) and Optos single-field fundus images and Clarus500 single-field fundus images (κ value=0.347, 95%CI 0.287-0.393) showed general agreement. ConclusionsCompared with Optos dual-field fundus image, dual-field 45°fundus image and Clarus500 dual-field fundus image have high consistency in the grading results of DR rapid screening. Compared with Optos single-field fundus image, the detection rate of the DR of Clarus500 single-field fundus image is higher.
ObjectiveTo analyze the change of differential genes and signaling pathways in high glucose induced BV2 cells, and to explore the mechanism of transgelin-2 (TAGLN2) regulating cellular inflammatory response and metabolic process. MethodsAn experimental study. The cultured BV2 cells were divided into mannitol treatment (Man) group, glucose treatment (Glu) group, overexpression control Glu treatment (Con) group, overexpression TAGLN2 Glu treatment group, silence control Glu treatment (shCon Glu) group, and silence TAGLN2 Glu treatment (shTAGLN2 Glu) group. Cells in the Man group were cultured in modified Eagle high glucose medium (DMEM) containing 25 mmol/L mannitol and 25 mmol/L glucose, cells in other groups (Glu group, Con Glu group, TAGLN2 Glu group, shCon Glu group and shTAGLN2 Glu group) were cultured in DMEM medium containing 50 mmol/L glucose. After 24 hours of cells culture, transcriptome sequencing of cells in each group were performed using high-throughput sequencing technology, and significantly differentially expressed genes (DEG) were screened. |log2 (fold change)|≥1 and P≤0.05 were adopted as criteria to screen for DEG. Gene Ontology (GO) function enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and protein-protein interaction network analysis were performed. Real-time polymerase chain reaction (RT-PCR) was used to detect the relative expression level of DEG mRNA. The data between groups were compared by independent sample t-test. ResultsWhen compared with Man group, a total of 517 differentially expressed genes were screened in Glu group, which including 277 up-regulated genes and 240 down-regulated genes. KEGG pathway enrichment analysis showed that the up-regulated genes were significantly enriched in immune system processes such as nuclear factor (NF)-κB signal pathway, Jak-signal transducers and activators of transcription (STAT) signal pathway, while down-regulated genes were significantly enriched in glycosaminoglycan degradation and glyceride metabolic pathway. Compared with Con Glu group, a total of 480 DEG were screened in TAGLN2 Glu group, among which 147 up-regulated and 333 down-regulated genes were detected. Up-regulated genes were significantly enriched in the metabolic processes of fatty acid, glyceride and pyruvate, while down-regulated genes were significantly enriched in immune system processes such as NF-κB signal pathway, Jak-STAT signal pathway and tumor necrosis factor (TNF) signal pathway. Compared with shCon Glu group, a total of 582 DEG were screened in shTAGLN2 Glu group, among which 423 up-regulated and 159 down-regulated genes were detected. Up-regulated DEG were significantly enriched in immune system processes such as TNF signal pathway and chemokine signal pathway, while down-regulated DEG were significantly enriched in pattern recognition receptor signal pathway. RT-PCR results showed that the relative expression levels of DEG mRNA Card11 (t=13.530), Icos (t=3.482), Chst3 (t=6.949), Kynu (t=5.399), interleukin (IL)-1β (t=2.960), TNF-α (t=5.800), IL-6(t=3.130), interferon-γ (t=7.690) and IL-17 (t=6.530) in the TAGLN2 Glu treatment group were decreased significantly compared with Con Glu group, and the difference was statistically significant. ConclusionTAGLN2 can inhibit glucose induced microglia inflammation by NF-κB and Jak-STAT signaling pathways, Card11, Icos, Chst3 and Kynu play an important role in the anti-inflammatory process of TAGLN2.
ObjectiveTo observe the efficacy of pars plana vitrectomy (PPV) in the treatment of different types of chorioretinal coloboma with retinal detachment (RD). MethodsA single-center, retrospective clinical study. From April 2021 to March 2023, 24 eyes of 23 patients who were diagnosed as chorioretinal coloboma with RD in Henan Provincial Eye Hospital were included in this study. There were 11 males with 12 eyes and 12 females with 12 eyes. The mean age was (33.3±13.7) years old. Best corrected visual acuity (BCVA), spectral domain optical coherence tomography were performed. The BCVA examination was performed using a international standard logarithmic visual acuity chart, which was converted into logarithm of the minimum angle of resolution (logMAR) visual acuity during statistics. According to the types of chorioretinal coloboma, the affected eyes were divided into the coloboma involved the optic disc group and the coloboma not involved the optic disc group, with 15 eyes and 9 eyes. According to whether the RD containing the coloboma area, the affected eyes were divided into RD containing the coloboma area group and the RD not containing the coloboma area group, with 15 eyes and 9 eyes. All eyes underwent standard pars plana three-channel 25G PPV, retinal laser photocoagulation combined with silicone oil tamponade. The follow-up time after surgery was (19.5±16.3) months. The last follow-up was the time point for efficacy determination. The retinal reattachment, BCVA recovery and postoperative complications were observed. Paired t-test or t test was performed for comparison of quantitative data. Fisher's exact test was performed for comparison of qualitative data. ResultsAt the last follow-up, retinal reattachment was achieved in 20 eyes (83.3%, 20/24). The logMAR BCVA of the coloboma involved the optic disc group before and after surgery were 1.85±0.62 and 1.71±0.71, the difference was no significant (t=0.845, P=0.412). The logMAR BCVA of the coloboma not involved the optic disc group before and after surgery were 1.75±0.45 and 0.84±0.26, the difference was statistically significant (t=6.153, P＜0.001). The improvement of BCVA in the coloboma not involved the optic disc group was significantly higher than that in the coloboma involved the optic disc group after surgery, with statistically significant differences (t=3.024, P=0.006). There was no significant difference in the retinal reattachment rate between the two groups (P=0.615). There was no significant difference in the retinal reattachment rate between the RD containing the coloboma area group and the RD not containing the coloboma area group (P=0.259). Postoperative complications included elevated intraocular pressure in five eyes, cataract progression in ten eyes, recurrent RD in two eyes, bullous keratopathy in one eye and band-shaped keratopathy in one eye. ConclusionPPV combined with silicone oil tamponade is safe and effective in the treatment of chorioretinal coloboma with RD, the improvement of visual acuity in the coloboma not involved the optic disc group is better than that in the coloboma involved the optic disc group after surgery.