Automatic Detection of 30 Fundus Diseases Using Ultra-Widefield Fluorescein Angiography with Deep Experts Aggregation.

INTRODUCTION: Inaccurate, untimely diagnoses of fundus diseases leads to vision-threatening complications and even blindness. We built a deep learning platform (DLP) for automatic detection of 30 fundus diseases using ultra-widefield fluorescein angiography (UWFFA) with deep experts aggregation. METHODS: This retrospective and cross-sectional database study included a total of 61,609 UWFFA images dating from 2016 to 2021, involving more than 3364 subjects in multiple centers across China. All subjects were divided into 30 different groups. The state-of-the-art convolutional neural network architecture, ConvNeXt, was chosen as the backbone to train and test the receiver operating characteristic curve (ROC) of the proposed system on test data and external test date. We compared the classification performance of the proposed system with that of ophthalmologists, including two retinal specialists. RESULTS: We built a DLP to analyze UWFFA, which can detect up to 30 fundus diseases, with a frequency-weighted average area under the receiver operating characteristic curve (AUC) of 0.940 in the primary test dataset and 0.954 in the external multi-hospital test dataset. The tool shows comparable accuracy with retina specialists in diagnosis and evaluation. CONCLUSIONS: This is the first study on a large-scale UWFFA dataset for multi-retina disease classification. We believe that our UWFFA DLP advances the diagnosis by artificial intelligence (AI) in various retinal diseases and would contribute to labor-saving and precision medicine especially in remote areas.

Relationship between ischemic index, leakage index, and macular edema in branch retinal vein occlusion.

PURPOSE: To investigate the combined association of the ischemic index and leakage index with macular edema on ultra-widefield fluorescein angiography (UWFFA) in patients with branch retinal vein occlusion (BRVO). METHODS: Retrospective image analysis study. The leakage index and ischemic index were calculated using Fiji after aligning early and late UWFFA images. Differences in the ischemic index, leakage index, and central macular thickness (CMT) between ischemic and non-ischemic BRVO were compared. Moreover, the association between the ischemic index, leakage index, and macular edema was analyzed. RESULTS: Eighty-three patients with BRVO were enrolled, including 53 non-ischemic BRVO and 30 ischemic BRVO patients. No significant differences were observed in leakage index and CMT between ischemic BRVO and non-ischemic BRVO (all P > 0.05). In all included patients, CMT correlated with the panretina and all subregion leakage indexes (all P < 0.01), but not with the ischemic index (all P > 0.05). In the ischemic BRVO group, CMT showed a correlation with the leakage index in several regions, but not with the ischemic index. After adjusting for the ischemic index and other clinical features, CMT remained significantly correlated with the leakage index in all regions. CONCLUSION: The leakage index may be a more effective biomarker for monitoring BRVO-associated macular edema compared to the ischemic index. Further follow-up studies are warranted to validate these findings.

Proliferative diabetic retinopathy and diabetic macular edema are two factors that increase macrophage-like cell density characterized by en face optical coherence tomography.

BACKGROUND: Macrophage-like cells (MLCs) located at the ILM were observed in live human retinas using adaptive optics optical coherence tomography (OCT) as well as clinically-used OCT. The study aimed to quantitatively analyzing MLCs at the vitreoretinal interface (VRI) in diabetic retinopathy (DR) using en face OCT and swept-source optical coherence tomography angiography (SS-OCTA). METHODS: 190 DR eyes were included in the study, with 70 proliferative diabetic retinopathy (PDR) eyes and 120 non- proliferative diabetic retinopathy (NPDR) eyes. Sixty-three eyes from normal subjects were included as controls. MLCs were visualized in a 5 µm en face OCT slab above the VRI centered on the fovea. Mann-Whitney U test and Kruskal-Wallis H test were used to compare the OCTA parameters and the MLC parameters among groups. We evaluated the MLC density among groups on binarized images after image processing. We also investigated the relationship between MLC density and other OCT parameters including retina thickness and vessel density (VD). RESULTS: The MLC density significantly increased in PDR eyes (PDR vs. NPDR, 8.97 (8.40) cells/mm2 vs.6.14 (8.78) cells/mm2, P = 0.013; PDR vs. normal, 8.97 (8.40) cells/mm2vs. 6.48 (6.71) cells/mm2, P = 0.027) and diabetic macular edema (DME) eyes (DME vs. without DME, 8.94 (8.26) vs.6.09 (9.00), P = 0.005). After adjusting for age and gender, MLC density in NPDR eyes negatively correlated to VD of deep capillary plexus (DCP) (P = 0.01). CONCLUSIONS: SS-OCTA is a non-invasive and simple method for the characterization of MLCs at the VRI. PDR and DME are two factors that increase MLC density. MLC density also correlated with VD.

Effects of Exosomal microRNAs on Oral Mucosal Epithelial Cells Cocultured with Limbal Niche Cells.

Autologous cultivated oral mucosal epithelial transplantation (COMET) is an important method for the treatment of limbal stem cell deficiency (LSCD), but the appearance of peripheral corneal neovascularization after COMET has prevented its widespread use in clinical practice. Using limbal niche cells (LNCs) as feeders in the process of coculturing could inhibit the postoperative corneal neovascularization. However, the specific mechanism is still unclear. In this study, LNCs were used as feeder cells to alter the phenotype of cultured oral mucosal epithelial cells (COMECs) by mimicking the primitive limbal microenvironment. The high-throughput sequencing of COMECs cocultured with LNCs or 3T3 cells (named LNCs group and 3T3 groups) was performed, and differential miRNA expression was analyzed. A total of 99 known and 1 newly predicted miRNAs were significantly upregulated in the LNCs group, while 101 known and 8 newly predicted miRNAs were significantly downregulated. A total of 3000 target genes with the 60 most significantly differentially expressed miRNAs were predicted, and 7 upregulated and 7 downregulated miRNAs were ultimately screened. The supernatants obtained from both cocultures were found to be rich in exosomes, indicating that the intercellular communication between COMECs and LNCs or 3T3 cells was highly active. Furthermore, the expression levels of rno-miR-200-5p, rno-miR-204-5p, rno-miR-126a-3p, rno-miR-192-5p, rno-miR-211-5p, rno-miR-143-3p, and rno-miR-184 were significantly higher in the LNCs group compared to the 3T3 group, and the expression levels had a similar trend in exosomes. Meanwhile, sequencing of the cell lines revealed 7 miRNAs that were significantly downregulated in the LNCs group. Interestingly, in that case, rno-miR-23a-3p, rno-miR-379-5p, and rno-miR-127-5p were also significantly downregulated in the exosomes. In summary, this study suggested that signal transduction between cells mediated by exosomal miRNAs may be an important factor for the inhibition of angiogenesis by LNCs nourished COMECs.

Incidence of and Risk Factors for Postoperative Hyphema After 23-Gauge Pars Plana Vitrectomy for Proliferative Diabetic Retinopathy.

PURPOSE: We aim to investigate the incidence, clinical course, and risk factors for developing postoperative hyphema after 23-gauge pars plana vitrectomy (PPV) for patients with proliferative diabetic retinopathy (PDR) without neovascularization of the iris or angles and neovascular glaucoma (NVG). METHODS: This retrospective study included 155 consecutive eyes from 124 patients with at least three-month follow-up who underwent PPV for PDR without neovascularization of the iris or angles and NVG. Demographic data, surgery notes, postoperative hyphema assessment, intraocular pressure (IOP), and the surgical outcome were recorded. RESULTS: Postoperative hyphema occurred in 18 of 155 eyes (11.6%), with 14 of those having hyphema on day 1, and 4 having hyphema on days 7-20. Of the 18 eyes, only 3 (16.7%) had normal IOP, and immediate intraocular hypertension was observed in 15 (83.3%). Seven eyes required anterior chamber paracentesis and five needed anterior chamber irrigation. The average time for absorption of the hyphema was 13.1 days, and IOP was controlled in all cases. There was a significant correlation between membrane removal and the development of hyphema (OR = 5.65 and 95% CI: 1.190-25.203; p = 0.013). No recurrence of hyphema was observed. In patients with hyphema, the final best corrected visual acuity (BCVA) was 1.75 ± 0.84 logMAR, which improved significantly compared to the initial BCVA of 2.20 ± 0.65 logMAR (t = 3.893; p = 0.001), and the final anatomic success rate was 100%. CONCLUSION: The development of hyphema is not uncommon after PPV for patients with PDR without neovascularization of the iris or angles and NVG, and membrane removal is a risk factor for postoperative hyphema. The timely management of hyphema ensures that hyphema does not affect the visual recovery or the final anatomical success.

Quantitative Assessment of Three-Dimensional Choroidal Vascularity and Choriocapillaris Flow Signal Voids in Myopic Patients Using SS-OCTA.

PURPOSE: To compare the choroidal vascularity of large- and middle-sized choroidal vessels and choriocapillaris (CC) perfusion in patients with different degrees of myopia using swept-source optical coherence tomography angiography (SS-OCTA). METHODS: One hundred and thirteen people with myopia were enrolled. SS-OCTA was performed to analyze the choroidal vascularity and CC perfusion. Three-dimensional (3D) choroidal vascularity index (CVI) and choroidal luminal volumes (LV) were obtained by artificial intelligence segmentation of the choroidal lumen in Volume OCT images. CC perfusion was assessed by flow signal voids (FSVs). RESULTS: In the macular, multiple linear regression model showed that choroidal thickness (CT), total choroidal volume, LV, and choroidal stromal volume were negatively correlated with axis length (AL), respectively (all p < 0.001). Three dimensional CVI was negatively associated with AL (p < 0.05). FSV% was positively correlated with age only (p < 0.001). Additionally, after adjustment for age and AL, FSV% had a significant negative correlation with CT (p < 0.05). CONCLUSION: Choroidal vascularity decreases gradually with increasing severity of myopia. The decrease of CC blood perfusion was related to a higher severity of myopia and the thinning of choroid.

NEW INSIGHTS INTO THE ASSOCIATION BETWEEN CHOROIDAL VESSELS AND CHORIOCAPILLARIS IN NORMAL EYES.

PURPOSE: To determine the relationship between large-sized and medium-sized choroidal vessels and choriocapillaris using the three-dimensional choroidal vascularity index (3D CVI) and percentage of flow deficits (FD%). METHODS: Prospective observational study. The main outcome measures included choroidal volume parameters (3D CVI, mean choroidal thickness, total choroidal volume, choroidal luminal volume, and choroidal stromal volume), choriocapillaris parameters (FD%, average area of FD, and FD count), and age and axial length. RESULTS: A total of 63 healthy individuals with an average age of 40.60 ± 12.32 years were included in this study. Univariate analysis showed that FD% (R = 0.295, P = 0.019) and area of FD (R = 0.276, P = 0.028) were significantly positively associated with 3D CVI, and FD count was negatively associated with 3D CVI (R = -0.297, P = 0.018). After adjusting for age and axial length, a significant correlation between FD% and 3D CVI (R = 0.264, P = 0.039) and between area of FD and 3D CVI remained (R = 0.267, P = 0.037). However, no significant correlation was found between FD count and 3D CVI after adjusting for axial length (R = -0.220, P = 0.085). CONCLUSION: In healthy individuals, choroidal vessel dilatation as suggested by elevated 3D CVI may lead to decreased choriocapillaris perfusion, which was correlated with an enlarged area of FD in choriocapillaris.

Expression of purinergic receptors on microglia in the animal model of choroidal neovascularisation.

To investigate the effect of P2 receptor on microglia and its inhibitor PPADS on choroidal neovascularization. Forty CX3CR1GFP/+ mice were randomly divided into 8 groups. In addition to the normal group, the rest of groups were receiving laser treatment. The retina and choroid from the second, third, fourth and fifth group of mice were taken in the 1, 4, 7, 14 days after laser treatment. The mice in the sixth and seventh group received intravitreal injection of 2 µl PPADS or PBS respectively immediately after laser treatment. The mice in the eighth group received topical application of PPADS once per day of three days. The mice in sixth, seventh and eighth group received AF and FFA examination on the fourth day after laser treatment. Immunofluorescence histochemical staining and real-time quantitative PCR were used to evaluate P2 expression and its effect on choroidal neovascularization. After laser treatment, activated microglia can express P2 receptors (P2X4, P2X7, P2Y2 and P2Y12). The expression of P2 increased on the first day after laser damage, peaked on the fourth day (tP2X4 = 6.05, tP2X7 = 2.95, tP2Y2 = 3.67, tP2Y12 = 5.98, all P < 0.01), and then decreased. After PPADS inhibition, compared with the PBS injection group, the mRNA of P2X4, P2X7, P2Y2 and P2Y12 were decreased significantly in the PPADS injection group (tP2X4 = 5.54, tP2X7 = 9.82, tP2Y2 = 3.86, tP2Y12 = 7.91, all P < 0.01) and the PPADS topical application group (tP2X4 = 3.24, tP2X7 = 5.89, tP2Y2 = 6.75, tP2Y12 = 4.97, all P < 0.01). Compared with the PBS injection group, not only the activity of microglia cells but also the leakage of CNV decreased significantly (P < 0.01) in the PPADS injection group and the PPADS topical application group. But between two PPADS groups, the leakage of CNV had no difference (P = 0.864). After laser induced CNV, activated microglia can express P2 receptors. The P2 receptor inhibitor, PPADS, can significantly affect the function of microglia and inhibit the formation of choroidal neovascularization.

Leakage index on ultra-widefield fluorescence angiography in different regions of retina and its correlation with cystoid macular edema in central retinal vein occlusion eyes.

PURPOSE: To investigate the relationship between leakage index on ultra-widefield fluorescence angiography (UWFFA) in different regions of retina and its correlation with cystoid macular edema (CME) in central retinal vein occlusion (CRVO) eyes. METHODS: Patients with naïve non-ischemic CRVO that had undergone UWFFA were identified. UWFFA images in the late phase were used to analyze the leakage index, which was performed by a semi-automatic method using ImageJ. The UWFFA images were subdivided into four regions by concentric circles centered on the macula for analysis. Optical coherence tomography (OCT) images were used to identify the presence of CME and obtain central macular thickness (CMT). RESULTS: A total of 57 eyes from 57 CRVO patients were analyzed in this study, including 43 eyes with CME and 14 eyes without CME. The leakage index in panretinal, peri-macular area (PMA), and near-peripheral area (NPA) was significantly different between eyes with CME and eyes without CME. Leakage index of PMA, NPA, mid-peripheral area (MPA), and panretinal area was significantly correlated with CMT, particularly the PMA. CONCLUSIONS: The distribution of leakage is different between patients with CME and patients without CME. The contribution of leakage index in different regions to CME was different, most predominant in PMA and NPA, and the closer to the center of the macula, the stronger the correlation between leakage index and CMT. A linear correlation was observed between CMT and the leakage index of panretinal area and all regions except far-peripheral area (FPA).

Observation of the far peripheral retina of normal eyes by ultra-wide field fluorescein angiography.

PURPOSE: The appearance of the far peripheral retina in normal eyes was observed using an Optos ultra-wide field fluorescein angiography imaging system. METHODS: A cross-sectional study of ultra-wide field pseudocolor and angiography images obtained using the Optos 200Tx. Four different non-vascular and six vascular-associated fluorescence features in far peripheral retina were analyzed. RESULTS: A total of 166 eyes in 83 patients met the inclusion criteria. The mean patient age was 40.78 ± 10.94 years (range: 20-65 years) with a female-to-male ratio of 0.98. Slight cataract were present in 64 eyes (38.55%), low to moderate myopia in 50 eyes (30.12%), and unspecified visual disturbances in 52 eyes (31.33%). Far peripheral retinas were found to have mottled florescent band in 42.17% eyes, ground glass hyperfluorescence in 84.34% eyes, granular hyperfluorescence in 12.65% eyes, local mottled fluorescence in 10.84% eyes, vascular anastomosis with a peripheral avascular area in 36.23% eyes, invisible capillary details in 50.67% eyes, vessels passing through the ora serrata in 16.98% eyes, late-phase slight leakage in 32.53% eyes, angiectasis in 18.67% eyes, and microaneurysm in 34.94% eyes. CONCLUSION: Ultra-wide field fluorescein angiography revealed various fluorescence features in the periphery of normal eyes, providing a basis for interpreting normal and pathological changes in peripheral retina and an important reference for clinical work.

Clinical application of ultra-widefield fundus autofluorescence.

PURPOSE: To review the basic principles of ultra-widefield fundus autofluorescence (UWF-FAF) and discuss its clinical application for a variety of retinal and choroidal disorders. METHODS: A systematic review of the PubMed database was performed using the search terms "ultra-widefield," "autofluorescence," "retinal disease" and "choroidal disease." RESULTS: UWF-FAF imaging is a recently developed noninvasive retinal imaging modality with a wide imaging range that can locate peripheral fundus lesions that traditional fundus autofluorescence cannot. Multiple commercially available ultra-widefield imaging systems, including Heidelberg Spectralis and Optomap Ultra-Widefield systems, are available to the clinician. Imaging by UWF-FAF is more comprehensive; it can reflect the content and distribution of the predominant ocular fluorophore in retinal pigment epithelial cells and evaluate the metabolic status of RPE of various retinal and choroidal disorders. CONCLUSION: UWF-FAF can detect abnormalities that traditional fundus autofluorescence cannot; therefore, it can be used to better elucidate disease pathogenesis, analyze genotype-phenotype correlations, diagnose and monitor disease.

Prospective associations of circulating adipocyte fatty acid-binding protein levels with risks of renal outcomes and mortality in type 2 diabetes.

AIMS/HYPOTHESIS: Elevated circulating adipocyte fatty acid-binding protein (AFABP) levels have been found to correlate with diabetic nephropathy staging in cross-sectional studies. However, it remains unclear whether these higher serum levels reflect a role of AFABP in the development of diabetic kidney disease (DKD), or simply result from its impaired renal clearance in DKD. Here we investigated prospectively the prognostic importance of serum AFABP level in the development of adverse renal outcomes in a large clinic-based cohort of participants with type 2 diabetes. METHODS: Baseline serum AFABP levels were measured in 5454 Chinese participants from the Hong Kong West Diabetes Registry. The association between circulating AFABP levels and incident adverse renal outcomes-defined as a composite endpoint of a sustained 40% decline in eGFR, end-stage renal disease requiring renal replacement therapy or kidney transplantation, or renal deaths-was evaluated using multivariable Cox regression analysis. RESULTS: Over a median follow-up of 5 years, 754 of the 5454 participants developed incident adverse renal outcomes. Elevated circulating AFABP levels were independently associated with incident adverse renal outcomes (HR 1.43, 95% CI 1.31, 1.57, p < 0.001) after adjustments for conventional risk factors for DKD progression. Importantly, the prognostic role of serum AFABP was independent of the baseline albuminuria status or eGFR levels of the study participants. CONCLUSIONS/INTERPRETATION: Circulating AFABP levels were predictive of incident adverse renal outcomes, even in participants with relatively well-preserved kidney function at baseline, suggesting its potential to be a useful marker for early risk stratification in DKD.

The multifaceted and controversial immunometabolic actions of adiponectin.

Adiponectin, one of the most abundant adipose-derived hormones, has variable actions in many tissues and organs. Although principally known for its insulin-sensitizing activity, recent data also highlight its homeostatic function, which is mediated both by direct actions on metabolic cells and indirectly through immunomodulatory effects on immune cells. Here we review the multifaceted immunometabolic actions of adiponectin and attempt to unify some of the contradictory reports on adiponectin function in inflammatory processes. We propose that a holistic understanding of adiponectin function can be garnered only from understanding its actions both on the immune system and on metabolism.

The roles of leptin and adiponectin: a novel paradigm in adipocytokine regulation of liver fibrosis and stellate cell biology.

Although leptin is a key adipokine promoting liver fibrosis, adiponectin may prevent liver injury. To determine the role of these adipokines in liver fibrosis and to understand their expression in vivo, fa/fa rats and their lean littermates were subjected to bile duct ligation (BDL). Histomorphometry for collagen and alpha-smooth muscle actin (alpha-SMA) revealed that lean rats, but not fa/fa littermates, had significant fibrosis with abundant hepatic stellate cell (HSC) activation. The lean-BDL rats had significantly higher leptin concentrations in the hepatic vein than lean sham-operated, fa/fa BDL, or fa/fa sham-operated rats. Co-localization of leptin and alpha-SMA in activated HSCs was observed by immunohistochemistry. Real-time reverse transcriptase-polymerase chain reaction and Western blot analysis confirmed the presence of leptin and alpha-SMA in activated, but not quiescent, HSCs, whereas only quiescent HSCs synthesized adiponectin mRNA and protein. Adiponectin overexpression in activated HSCs reduced proliferation, augmented apoptosis, and reduced expression of alpha-SMA and proliferating cell nuclear antigen. Adiponectin receptors (AdipoR1 and AdipoR2) were detected in both activated and quiescent HSCs, but only activated HSCs produced significant apoptosis after treatment with either globular or full-length adiponectin. Adiponectin may act to reverse HSC activation, maintain HSC quiescence, or significantly, may have important therapeutic implications in liver fibrosis.