Role of immune inflammation regulated by macrophage in the pathogenesis of age-related macular degeneration.

Age-related macular degeneration (AMD) can lead to irreversible impairment of visual function, and the number of patients with AMD has been increasing globally. The immunoinflammatory theory is an important pathogenic mechanism of AMD, with macrophages serving as the primary inflammatory infiltrating cells in AMD lesions. Its powerful immunoinflammatory regulatory function has attracted considerable attention. Herein, we provide an overview of the involvement of macrophage-regulated immunoinflammation in different stages of AMD. Additionally, we summarize novel therapeutic approaches for AMD, focusing on targeting macrophages, such as macrophage/microglia modulators, reduction of macrophage aggregation in the subretinal space, modulation of macrophage effector function, macrophage phenotypic alterations, and novel biomimetic nanocomposites development based on macrophage-associated functional properties. We aimed to provide a basis and reference for the further exploration of AMD pathogenesis, developmental influences, and new therapeutic approaches.

ERp29 Attenuates Nicotine-Induced Endoplasmic Reticulum Stress and Inhibits Choroidal Neovascularization.

Nicotine-induced endoplasmic reticulum (ER) stress in retinal pigment epithelium (RPE) cells is thought to be one pathological mechanism underlying age-related macular degeneration (AMD). ERp29 attenuates tobacco extract-induced ER stress and mitigates tight junction damage in RPE cells. Herein, we aimed to further investigate the role of ERp29 in nicotine-induced ER stress and choroidal neovascularization (CNV). We found that the expression of ERp29 and GRP78 in ARPE-19 cells was increased in response to nicotine exposure. Overexpression of ERp29 decreased the levels of GRP78 and the C/EBP homologous protein (CHOP). Knockdown of ERp29 increased the levels of GRP78 and CHOP while reducing the viability of ARPE-19 cells under nicotine exposure conditions. In the ARPE-19 cell/macrophage coculture system, overexpression of ERp29 decreased the levels of M2 markers and increased the levels of M1 markers. The viability, migration and tube formation of human umbilical vein endothelial cells (HUVECs) were inhibited by conditioned medium from the ERp29-overexpressing group. Moreover, overexpression of ERp29 inhibits the activity and growth of CNV in mice exposed to nicotine in vivo. Taken together, our results revealed that ERp29 attenuated nicotine-induced ER stress, regulated macrophage polarization and inhibited CNV.

Comparison of functional changes of retina after subthreshold and threshold pan-retinal photocoagulation in severe non-proliferative diabetic retinopathy.

PURPOSE: To find a new approach of pan-retinal photocoagulation (PRP) with less damage to the retina in the treatment of severe non-proliferative diabetic retinopathy (NPDR), this study compared functional changes in the retina after subthreshold and threshold PRP treatment in severe NPDR eyes. METHODS: Post hoc analysis of a randomized clinical trial was conducted in this study. Seventy eyes of 35 patients with bilateral, symmetric, severe NPDR were enrolled. Two eyes from the same patient were randomized into two groups, one eye received subthreshold PRP (S-PRP) and the other eye received threshold PRP (T-PRP). Comprehensive ophthalmological evaluations were performed on the baseline and every 3 months for 1 year. Visual field (VF) and full-field electroretinography (ERG) were performed on the baseline and repeated at month 12. RESULTS: During the 12-month follow-up, 4 eyes (11.4%) in the S-PRP group and 3 eyes (8.6%) in the T-PRP group progressed to proliferative diabetic retinopathy (PDR) stage, and there was no statistical difference in PDR progression rate between the two groups (P = 0.69). In addition, the changes in best-corrected visual acuity (BCVA) from baseline to month 12 between the two groups had no statistical difference (P = 0.30). From baseline to month 12, changes in central VF between the two groups had no statistical difference (P = 0.25), but changes in total score points of peripheral VF in the S-PRP group (- 242.1 ± 210.8 dB) and the T-PRP group (- 308.9 ± 209.7 dB) were statistically significant (P = 0.03). At month 12, ERG records showed that the amplitude of dark-adapted 0.01 ERG, dark-adapted 3.0 ERG, oscillatory potentials, light-adapted 3.0 ERG, and 30 Hz flicker ERG of both groups were significantly decreased from the baseline (P < 0.05). In addition, the amplitude of each ERG record in the S-PRP group decreased significantly less than those in the T-PRP group (P < 0.05). CONCLUSIONS: Subthreshold PRP is as effective as threshold PRP for preventing severe NPDR progress to PDR within 1 year with less damage to periphery VF and retinal function. CLINICALTRIALS: gov Identifier: NCT01759121.

Determinants of macular ganglion cell-inner plexiform layer thickness in normal Chinese adults.

BACKGROUND: Demographic, systemic and ocular factors may impact macular ganglion cell-inner plexiform layer (GCIPL) thickness measurements. This study aimed to investigate the influences of multiple potential determinants of macular GCIPL thickness in normal Chinese adults. METHODS: This was a retrospective study conducted on 225 normal eyes from 225 healthy Chinese adults. GCIPL thickness were obtained using Cirrus high-definition optical coherence tomography (OCT). The age, gender, laterality, spherical equivalent (SE) refractive error, intraocular pressure (IOP), axial length (AL), central cornea thickness (CCT), circumpapillary retinal nerve fibre layer (pRNFL) thickness and OCT signal strength were recorded and their respective effect on GCIPL thickness parameters were evaluated. RESULTS: The mean (± SD) average, minimum, superotemporal, superior, superonasal, inferonasal, inferior, and inferotemporal GCIPL thickness was 84.56 ± 5.36, 81.32 ± 5.58, 83.08 ± 5.37, 85.70 ± 5.95, 87.15 ± 6.26, 85.07 ± 6.11, 82.46 ± 5.76, and 83.88 ± 5.59 µm, respectively. Determinants of thinner GCIPL thickness were older age (P = 0.001-0.117; effects enhanced if age over 40 years), thinner pRNFL (all P < 0.001), and weaker signal strength (all P < 0.001). No significant difference was found between males and females (P = 0.069-0.842), and between right eyes and the left eyes (P = 0.160-0.875) except that of superonasal GCIPL thickness (P < 0.001). There was no significant correlation between GCIPL thickness and SE, IOP, CCT, and AL (P = 0.135-0.968). CONCLUSIONS: Individual determinants associated with thinner GCIPL thickness were older age (particularly over 40 years of age), thinner pRNFL, and weaker OCT signal strength. This is relevant in comprehensively understanding the normative data and differentiating normal aging from abnormalities.

Subthreshold Pan-Retinal Photocoagulation Using Endpoint Management Algorithm for Severe Nonproliferative Diabetic Retinopathy: A Paired Controlled Pilot Prospective Study.

PURPOSE: The aim of this study was to report the efficacy and safety profile of subthreshold pan-retinal photocoagulation (PRP) using endpoint management (EPM) algorithm compared with conventional threshold PASCAL PRP for the treatment of severe nonproliferative diabetic retinopathy (NPDR). METHODS: This was a prospective, single-center, paired randomized controlled trial of 56 eyes of 28 participants with bilateral symmetric severe NPDR. One eye of the participant was randomly assigned to receive the subthreshold EPM PRP, while the other eye of the same participant received the threshold PASCAL PRP. The primary outcome measures included the difference in the 1-year risk of progression to PDR between 2 groups, and mean changes of the logarithm of the minimal angle of resolution (logMAR) visual acuity (VA). The second outcome measures included central foveal thickness (CFT), 1-year risk of progression to PDR, and visual field (VF) parameters. RESULTS: The subthreshold EPM PRP group and the threshold PASCAL PRP group had similar 1-year risk of progression to PDR during the 12-month follow-up visits (17.86 vs. 14.29%, p > 0.05). Slightly decreased VA was found in both groups (0.08 vs. 0.09 logMAR VA); however, no statistical difference was found for neither group (p > 0.05). Similar results were found for thickened CFT for both groups (23.59 vs. 28.34 µm, p > 0.05). Specifically, although substantial loss of VF was found in the threshold PASCAL PRP group (p < 0.05), no obvious damage to VF was seen in the subthreshold EPM PRP group (p > 0.05). CONCLUSION: The subthreshold EPM PRP is noninferior to the conventional threshold PASCAL PRP in the treatment of severe NPDR during 12-month follow-up and could be an alternative treatment option for patients with severe NPDR.

Dynamic changes and correlation analysis of outer retinal microstructure in macular area of central serous chorioretinopathy patients during restoration period.

PURPOSE: To investigate the dynamic changes and possible affecting variables of outer retinal microstructure in macular area of central serous chorioretinopathy (CSC) patients. METHODS: This was a retrospective study. The data of optical coherence tomography (OCT) and autofluoroscopy (AF) of 36 CSC patients admitted to our hospital from February 2012 to February 2013 were included. Dynamic variations and possible correlated variables of central retinal thickness (CRT), subretinal fluid diameter (SRFD), ellipsoid zone (EZ), interdigitation zone (IZ) and/or hyperautofluorescent spot (HAS) were analyzed. RESULTS: The outer retinal microstructure was gradually restored along with the subretinal fluid absorption during the follow-up. EZ in 94.4% (34/36) and the IZ in 100% (36/36) eyes were completely disappeared at baseline and restored (completed or incomplete) in 88.9% (8/9) and 44.4% (4/9) eyes, respectively, after 6-month follow-up. HAS was evident in 25% eyes (8/32 eyes) at baseline, and the density was initially increased and then declined during follow-up. Correlation analysis demonstrated that the restoration of EZ and IZ was correlated with the restoration period and subretinal fluid absorption. CONCLUSION: The outer retinal microstructure was restored during the subretinal fluid absorption in CSC patients, with EZ restored earlier than IZ. The restoration period and the absorption of subretinal fluid were two closely correlated variables of macular microstructure restoration.

Anatomical and functional responses in eyes with diabetic macular edema treated with "1 + PRN" ranibizumab: one-year outcomes in population of mainland China.

BACKGROUND: To evaluate the anatomical and functional responses in eyes with diabetic macular edema (DME) treated with ranibizumab under "1 + pro re nata (PRN)" regimen. METHODS: This prospective interventional case series included 69 eyes of 69 patients with DME treated with intravitreal injections of 0.5 mg ranibizumab followed by repeated injections as needed. Best-corrected visual acuity (BCVA), central foveal thickness (CFT), subfoveal choroidal thickness (SFCT), and predictive factors for final visual outcomes were assessed. RESULTS: Logarithm of minimal angle of resolution (logMAR) BCVA improved from 0.64 ± 0.23 at baseline to 0.56 ± 0.27, 0.53 ± 0.26, 0.47 ± 0.25, 0.44 ± 0.32, 0.47 ± 0.26 and 0.46 ± 0.26 at time-point of months 1, 2, 3, 6, 9, and 12, respectively (P < 0.05 for any follow-up time-point except month 1). CFT decreased from 478.23 ± 172.31 µm at baseline to 349.74 ± 82.21 µm, 313.52 ± 69.62 µm, 292.59 ± 61.07 µm, 284.67 ± 69.85 µm, 268.33 ± 43.03 µm, and 270.39 ± 49.27 µm at above time-points, respectively (P < 0.05). The number of injections was 6.83 times over 12 months' follow-up under "1 + PRN" regimen. Multivariate analysis showed that the factors including age, BCVA at baseline, disruption of ellipsoid zone, posterior vitreous detachment (PVD), and vitreomacular traction (VMT) were correlated with the final BCVA. CONCLUSIONS: Intravitreal injections of ranibizumab under "1 + PRN" regimen is a not only effective but also safe way to improve visual acuity of DME patients. And older age, lower baseline BCVA, VMT, and disruption of ellipsoid zone are predictors for final poor BCVA while PVD is a positive predictive factor for good final BCVA. TRIAL REGISTRATION: The trial was registered retrospectively in ClinicalTrials.gov on 2 June 2019 (NCT03973138).

Effectiveness and Safety of Intravitreal Injection of Conbercept as an Initial Treatment for Exudative Circumscribed Choroidal Hemangioma.

OBJECTIVE: To investigate the effectiveness and safety of intravitreal injection of conbercept (IVC) as the initial treatment for exudative circumscribed choroidal hemangioma (CCH). METHODS: Forty-two eyes of 42 patients received 3 monthly IVC (0.5 mg/0.05 mL) as the initial treatment. Three months later, the patients were assessed for further treatment including observation, reinjection of conbercept, laser photocoagulation (if the lesion was 3,000 µm away from the macular fovea), or photodynamic therapy (PDT; if the lesion was under the macular fovea). Anatomical and functional responses including best corrected visual acuity (BCVA), central foveal thickness (CFT), and tumor size were analyzed. RESULTS: Twenty-three patients (54.76%) were sensitive to the monotherapy of IVC. Fourteen patients (33.33%) were insensitive to IVC and underwent rescue laser photocoagulation, and 5 patients (11.90%) underwent rescue PDT due to insensitivity to IVC treatment at 3 months. For subgroup analysis, although no statistical difference was found for BCVA at any follow-up time point compared to baseline, an increasing tendency of BCVA was found in the IVC group (p> 0.05). The mean CFT decreased significantly from 427.13 ± 214.74 µm at baseline to 259.83 ± 61.68 µm at 6 months in the IVC group (p< 0.05). No influence on tumor size was found in the IVC group. CONCLUSION: IVC as the initial treatment might be an option for exudative CCH.

APRPG-modified nanoliposome loaded with miR-146a-5p inhibitor suppressed choroidal neovascularization by targeting endothelial cells.

INTRODUCTION: To explore the effect and mechanism of APRPG-modified nanoliposomes loaded with miR-146a-5p inhibitor (ANL-miR-146a-5p inhibitor) on endothelial cell proliferation, migration, tube formation, and choroidal neovascularization (CNV) in mice. METHODS: ANL-miR-146a-5p inhibitors were generated by thin film hydration; in vitro, endothelial cell uptake experiment was used to detect the targeting effect of ANL-miR-146a-5p inhibitor; endothelial cells proliferation, migration, and tube formation were detected, respectively, by CCK8 assay, scratch assay, and Matrigel tube formation assay. In vivo, the mice CNV models were established by 810 nm laser photocoagulation. Mice choroidal flatmounts were performed to detect the volume of CNV after intravitreal injection of L-miR-146a-5p inhibitor, ANL-miR-146a-5p inhibitor, or normal saline; the vascular endothelial growth factor (VEGF) expression of mice choroidal tissue was detected by ELISA; HE section and electrophysiology (ERG) were performed to check the toxicity of ANL-miR-146a-5p inhibitor on mice retina. RESULTS: ANL are targeted to endothelial cells and are more targeted in inflammatory environment. At the same concentration, ANL-miR-146a-5p inhibitor's ability to inhibit endothelial cell proliferation, migration, tube formation, CNV formation, and VEGF expression is better than L-miR-146a-5p inhibitor (p < 0.05); ANL-miR-146a-5p inhibitor had no toxicity on the structure of mouse retina; ANL-miR-146a-5p inhibitor had no toxicity on the a-wave and b-wave amplitudes and b-wave implicit times (p > 0.05). CONCLUSIONS: ANL-miR-146a-5p inhibitor can more effectively down-regulate the expression level of VEGF through its targeting to endothelial cells, thereby more effectively inhibiting endothelial cell proliferation, migration, tube formation, and mice CNV formation.

A pilot prospective study of 577-nm yellow subthreshold micropulse laser treatment with two different power settings for acute central serous chorioretinopathy.

To compare the efficacy of 50% threshold power with 25% threshold power of 577-nm subthreshold micropulse laser (SMPL) for acute central serous chorioretinopathy (CSC). Prospective, interventional, non-randomized, comparative case series. A total of 54 patients (54 eyes) with acute CSC were enrolled. Twenty-four eyes received 25% threshold power and 30 eyes received 50% threshold power of 577-nm SMPL. Best-corrected visual acuity (BCVA), central macular thickness (CMT), and complete absorption of subretinal fluid (SRF) were evaluated at 1 month and 3 months. The complete absorption rate of SRF in the 50% power group was significantly greater than that in the 25% power group at 1 month (70.0% vs 25.0%, p < 0.001) and at 3 months (83.3% vs 54.2%, p < 0.001). Mean BCVA improved from 0.34 ± 0.20 LogMAR to 0.02 ± 0.13 LogMAR in the 50% power group and from 0.27 ± 0.15 LogMAR to 0.14 ± 0.21 LogMAR in the 25% power group with a significant difference between the two groups after 3 months (p = 0.027). In the 50% power group, the CMT decreased from 491.6 ± 154.8 µm at baseline to 231.3 ± 92.3 µm at 1 month and 228.2 ± 88.1 µm at 3 months, and in the 25% power group, the CMT decreased from 444.9 ± 164.1 to 306.8 ± 102.6 µm at 1 month and 254.5 ± 101.7 µm at 3 months. There was statistical difference of CMT at 1 month (p = 0.009) but no significant difference at 3 months between the two groups (p = 0.232). SMPL with 50% threshold power may be more effective than 25% threshold power for acute CSC.

Comparison of the effects of photodynamic therapy, intravitreal ranibizumab and combination for polypoidal choroidal vasculopathy under 1 + PRN regimen.

BACKGROUND: The optimal treatment for polypoidal choroidal vasculopathy (PCV) is still under debate. Little knowledge is known about the treatment effect of "1+pro re nata(PRN)" treatment regimen for PCV. The aim of this study was to compare the outcomes of photodynamic therapy (PDT), intravitreal ranibizumab injection (IVR) and combination therapy under the "1 + PRN" treatment regimen for PCV. METHODS: Fifty-seven eyes of 57 patients completed the 12 months' follow-up in this prospective study. The patients in the PDT arm(n = 23), ranibizumab arm(n = 18), or combination arm(n = 16) underwent a session of PDT, IVR or combination of both at baseline followed by additional IVR as needed. Mean change of logarithm of the minimal angle of resolution (logMAR) visual acuity (VA), central foveal thickness (CFT) and the regression rate of polyps were evaluated. Cost-benefit analysis was also performed. RESULTS: At Month 12, the mean logMAR VA improved from 0.90 ± 0.52 to 0.75 ± 0.57 in the PDT group (P < 0.05), from 0.96 ± 0.58 to 0.77 ± 0.41 in the IVR group (P < 0.05), and from 0.94 ± 0.55 to 0.72 ± 0.44 in the combination group (P < 0.05), respectively. The CFT decreased from 478.04 ± 156.70 µm, 527.5 ± 195.90 µm, and 522.63 ± 288.40 µm at the baseline to 366.43 ± 148.28 µm, 373.17 ± 134.88 µm and 328.44 ± 103.25 in the PDT group (P < 0.05), IVR group (P < 0.01), and the combination group (P < 0.05), respectively. However, no statistical difference was found between groups (P > 0.05). PDT treatment (60.87%) was superior to the IVR therapy (22.22%) in achieving complete regression of polyps (P < 0.05). Cost-benefit analysis showed that IVR treatment cost the least money for improving per 0.1logMAR units and the combination therapy demanded the least money for reducing per 100 µm of CFT. CONCLUSIONS: PDT, IVR and the combination therapy have similar efficacy in the VA improvement as well as the reduction of CFT under the "1 + PRN" treatment regimen. TRIAL REGISTRATION: Current Controlled Trials NCT03459144 . Registered retrospectively on March 2, 2018.

Functional and morphological analysis of the subretinal injection of human retinal progenitor cells under Cyclosporin A treatment.

PURPOSE: The purpose of this study is to evaluate the functional and morphological changes in subretinal xenografts of human retinal progenitor cells (hRPCs) in B6 mice treated with Cyclosporin A (CsA; 210 mg/l in drinking water). METHODS: The hRPCs from human fetal eyes were isolated and expanded for transplantation. These cells, with green fluorescent protein (GFP) at 11 passages, were transplanted into the subretinal space in B6 mice. A combination of invasive and noninvasive approaches was used to analyze the structural and functional consequences of the subretinal injection of the hRPCs. The process of change was monitored using spectral domain optical coherence tomography (SDOCT), histology, and electroretinography (ERG) at 3 days, 1 week, and 3 weeks after transplantation. Cell counts were used to evaluate the survival rate with a confocal microscope. ERGs were performed to evaluate the physiologic changes, and the structural changes were evaluated using SDOCT and histological examination. RESULTS: The results of the histological examination showed that the hRPCs gained a better survival rate in the mice treated with CsA. The SDOCT showed that the bleb size of the retinal detachment was significantly decreased, and the retinal reattachment was nearly complete by 3 weeks. The ERG response amplitudes in the CsA group were less decreased after the injection, when compared with the control group, in the dark-adapted and light-adapted conditions. However, the cone-mediated function in both groups was less affected by the transplantation after 3 weeks than the rod-mediated function. CONCLUSIONS: Although significant functional and structural recovery was observed after the subretinal injection of the hRPCs, the effectiveness of CsA in xenotransplantation may be a novel and potential approach for increasing retinal progenitor cell survival.

Intravitreal injection of (99)Tc-MDP inhibits the development of laser-induced choroidal neovascularization in rhesus monkeys.

BACKGROUND: The aim of this study was to investigate the safety and efficacy of intravitreal injection of (99)Tc-MDP, a decay product of (99m)Tc-MDP, on the development of laser-induced choroidal neovascularization (CNV) in rhesus monkeys. METHODS: Experimental CNV was induced by argon laser with a small high-energy laser spot. Monkeys were given 50 µL of (99)Tc-MDP at a concentration of 0.005 µg/mL (n = 6) or 0.01 µg/mL (n = 6) by intravitreal injection once a week immediately after laser injury for a period of 56 days. Control animals were treated with the same volume of PBS (n = 6) in the same way. Eyes were monitored by ophthalmic examination, color fundus photography, fluorescence fundus angiography (FFA), optical coherence tomography (OCT) and histology. Incidences of grade 4 CNV lesions as well as the leakage areas of grade 4 CNVs on the late-phase of fluorescein angiograms were measured in a standardized, randomized and masked fashion fortnightly. The maximum widths and heights of grade 4 CNVs were also calculated by histology at the end of the experiment. Toxicity of (99)Tc-MDP on the retina was evaluated by electroretinogram (ERG) and histologic analysis. RESULTS: (99)Tc-MDP reduced the incidences of grade 4 CNVs by 33.33 % and 39.40 % in the 0.005 µg/mL and 0.01 µg/mL groups, respectively, compared with the PBS group on day 28 (P < 0.05; n = 6). The leakage areas of grade 4 CNVs were smaller in the 0.005 µg/mL (0.7136 ± 0.0283 mm(2), p <0.01; n = 6) and 0.01 µg/mL (0.4351 ± 0.0349 mm(2), p < 0.01; n = 6) groups than those in the PBS control group (0.9373 ± 0.0455 mm(2); n = 6) in a dose-dependent manner on day 28. OCT and histology also showed that the sizes of CNVs were smaller in the (99)Tc-MDP treated groups than those in the PBS group. Although intravitreal injection of (99)Tc-MDP led to mild inflammatory reaction in the anterior chamber, histology and ERG findings demonstrated that (99)Tc-MDP did not cause any change in histological structure or function of the retina (p>0.05). CONCLUSIONS: Intravitreal injection of (99)Tc-MDP can inhibit the development of laser-induced CNV without toxic effect on retina, suggesting that (99)Tc-MDP has therapeutic potential for CNV related diseases.

[The study of laser-induced choroidal neovascularization in rhesus monkeys].

OBJECTIVE: To investigate the morphological and functional changes of the laser-induced choroidal neovascularization (CNV) in rhesus monkeys. METHODS: Experimental study. Eight adult rhesus monkeys weighted 4 to 7 kg were used in this study. CNVs were induced with small high-energy laser spots at short pulse duration by an argon green laser. Eyes were monitored weekly by color fundus photography, fluorescence fundus angiography (FFA) , and optical coherence tomography (OCT) . Fluorescein leaking intensities of grade 4 CNV lesions were analyzed by the method of ANOVA for repeated measures. Electroretinogram (ERG) was performed before laser photocoagulation and 56 days after laser photocoagulation and the data were analyzed with paired t-test. RESULTS: (1) FFA revealed that the mean intensities of grade 4 CNV lesions were 89.44 ± 26.28, 97.56 ± 26.47, 110.22 ± 29.76, 100.26 ± 29.24, 91.77 ± 28.11, 77.76 ± 24.85 and 63.23 ± 22.34 on day 14, day 21, day 28, day 35, day 42, day 49, and day 56 respectively and the differences were statistically significant (F = 39.715, P < 0.01) . The differences between any time-point and its previous time-point were also statistically significant (t14-21 = 4.824, P < 0.01; t21-28 = 5.225, P < 0.01; t28-35 = 7.378, P < 0.01;t35-42 = 2.954, P < 0.05; t42-49 = 5.386, P < 0.01; t49-56 = 6.138, P < 0.01). (2) OCT images showed retinal edema, subretinal fluid and hyper-reflective lesions of CNVs in the laser sites and histopathology showed that fibrovascular tissues together with proliferating retinal pigment epithelium cells were seen in the laser sites. (3) ERG data revealed that implicit time of dark-adapted b wave (t = 4.23, P < 0.01) increased while the amplitudes of dark-adapted a wave (t = 6.35, P < 0.01) , dark-adapted b wave (t = 3.12, P < 0.01) and light-adapted b wave (t = 3.93, P < 0.01) decreased 56 days after laser photocoagulation compared with those before laser photocoagulation. CONCLUSION: The laser-induced CNV in non-human primate model shows continuous leakage on FFA examination and reduced amplitudes as well as increased implicit time on ERG examination, suggesting that laser-induced CNV primate model not only could be used to study the pathogenesis of CNV formation but also could be used for screening of drug effectiveness.

PEDF protects retinal pigment epithelium from ferroptosis and ameliorates dry AMD-like pathology in a murine model.

Age-related macular degeneration (AMD) is the leading cause of irreversible vision damage among elderly individuals. There is still no efficient treatment for dry AMD. Retinal pigment epithelial (RPE) degeneration has been confirmed to play an important role in dry AMD. Recent studies have reported that ferroptosis caused by iron overload and lipid peroxidation may be the primary causes of RPE degeneration. However, the upstream regulatory molecules of RPE ferroptosis remain largely unknown. Pigment epithelium-derived factor (PEDF) is an important endogenic protective factor for the RPE. Our results showed that in the murine dry AMD model induced by sodium iodate (SI), PEDF expression was downregulated. Moreover, dry AMD-like pathology was observed in PEDF-knockout mice. Therefore, the aim of this study was to reveal the effects and mechanism of PEDF on RPE ferroptosis and investigate potential therapeutic targets for dry AMD. The results of lipid peroxidation and transmission electron microscope showed that retinal ferroptosis was significantly activated in SI-treated mice and PEDF-knockout mice. Restoration of PEDF expression ameliorated SI-induced retinal dysfunction in mice, as assessed by electroretinography and optical coherence tomography. Mechanistically, western blotting and immunofluorescence analysis demonstrated that the overexpression of PEDF could upregulate the expression of glutathione peroxidase 4 (GPX4) and ferritin heavy chain-1 (FTH1), which proved to inhibit lipid peroxidation and RPE ferroptosis induced by SI. This study revealed the novel role of PEDF in ferroptosis inhibition and indicated that PEDF might be a potential therapeutic target for dry AMD.

The RBPMSCreERT2-tdTomato mouse line for studying retinal and vascular relevant diseases.

RNA-binding protein with multiple splicing (RBPMS) plays a crucial role in cardiac mesoderm specification and cardiovascular development, as well as being a typical marker for whole retinal ganglion cells (RGCs). However, there is a lack of animal models to spatiotemporally trace the location and function of RBPMS-expressing cells in vivo. In this study, we develop a tamoxifen-inducible RBPMS-tdTomato reporter mouse line to track RBPMS-expressing cells during embryogenesis and adulthood. This mouse line allows us to identify and locate RBPMS-tdTomato-positive cells among various tissues, especially in RGCs and smooth muscle cells, which assist to simulate related retinal degenerative diseases, model and examine choroidal neovascularization non-invasively in vivo. Our results show that the RBPMSCreERT2-tdTomato mouse line is a valuable tool for lineage tracing, disease modeling, drug screening, as well as isolating specific target cells.

Systemic Risk Factors for Vitreous Hemorrhage Secondary to Polypoidal Choroidal Vasculopathy.

INTRODUCTION: It remains unclear whether systemic factors are associated with an increased risk of vitreous hemorrhage (VH) secondary to polypoidal choroidal vasculopathy (PCV), and there is no method to predict the possibility of VH occurrence in patients with PCV. This study aimed to investigate and visualize systemic risk factors for VH in patients with PCV. METHODS: Data on the sex, age, history of systematic diseases, best-corrected visual acuity, intraocular pressure, and laboratory data of patients with PCV were collected from the medical record system. Univariate and multivariate binary logistic regression analyses were applied to investigate independent risk factors for VH in patients with PCV. Receiver operating characteristic analysis and nomograms were used to visualize the independent risk factors. RESULTS: The patient population comprised 115 patients with VH secondary to PCV and 181 patients with PCV without VH. Binary logistic regression analyses showed that higher white blood cell count [WBC; odds ratios (OR) 1.247], higher aspartate aminotransferase/alanine aminotransferase ratio (AST/ALT; OR 2.339), and longer activated partial thromboplastin time (APTT; OR 1.196) were independent risk factors of VH in patients with PCV. Integrated application of APTT, AST/ALT, and WBC as markers showed the best performance for distinguishing patients with VH, with an area under the curve of 0.723. The nomogram was created for doctors to calculate the possibility of VH in a patient with PCV. CONCLUSIONS: Higher WBC, higher AST/ALT, and longer APTT are independent serum risk factors of VH secondary to PCV, which may shed light on VH prevention in patients with PCV.

Novel compound heterozygous variants of the SEC23A gene in a Chinese family with cranio-lenticulo-sutural dysplasia based on data from a large cohort of congenital cataract patients.

BACKGROUND: Cranio-lenticulo-sutural dysplasia (CLSD) is a rare dysmorphic syndrome characterized by skeletal dysmorphism, late-closing fontanels, and cataracts. CLSD is caused by mutations in the SEC23A gene (OMIM# 607812) and can be inherited in either an autosomal dominant or autosomal recessive pattern. To date, only four mutations have been reported to cause CLSD. This study aims to identify the disease-causing variants in a large cohort of congenital cataract patients, to expand the genotypic and phenotypic spectrum of CLSD, and to confirm the association between SEC23A and autosomal recessive CLSD (ARCLSD). METHODS: We collected detailed medical records and performed comprehensive ocular examinations and whole-exome sequencing (WES) on 115 patients with congenital cataracts. After suspecting that a patient may have CLSD based on the sequencing results, we proceeded to conduct transmission electron microscopy (TEM) on the cultured skin fibroblasts. The clinical validity of the reported gene-disease relationships for the gene and the disease was evaluated using the ClinGen gene curation framework. RESULTS: Two novel compound heterozygous variants (c.710A > C p.Asp237Ala, c.1946T > C p.Leu649Pro) of the SEC23A gene, classified as variant of uncertain significance, were identified in the proband with skeletal, cardiac, ocular, and hearing defects. The observation of typical distended endoplasmic reticulum cisternae further supported the diagnosis of CLSD. Application of the ClinGen gene curation framework confirmed the association between SEC23A and ARCLSD. CONCLUSION: This study expands the genotypic and phenotypic spectrum of CLSD, proposes TEM as a supplemental diagnostic method, and indicates that congenital cataracts are a typical sign of ARCLSD.

Performance of the AIDRScreening system in detecting diabetic retinopathy in the fundus photographs of Chinese patients: a prospective, multicenter, clinical study.

Background: Diabetic retinopathy (DR) is the leading cause of blindness in the working-age population worldwide, and there is a large unmet need for DR screening in China. This observational, prospective, multicenter, gold standard-controlled study sought to evaluate the effectiveness and safety of the AIDRScreening system (v. 1.0), which is an artificial intelligence (AI)-enabled system that detects DR in the Chinese population based on fundus photographs. Methods: Participants with diabetes mellitus (DM) were recruited. Fundus photographs (field 1 and field 2) of 1 eye in each participant were graded by the AIDRScreening system (v. 1.0) to detect referable DR (RDR). The results were compared to those of the masked manual grading (gold standard) system by the Zhongshan Image Reading Center. The primary outcomes were the sensitivity and specificity of the AIDRScreening system in detecting RDR. The other outcomes evaluated included the system's diagnostic accuracy, positive predictive value, negative predictive value, diagnostic accuracy gain rate, and average diagnostic time gain rate. Results: Among the 1,001 enrolled participants with DM, 962 (96.1%) were included in the final analyses. The participants had a median age of 60.61 years (range: 20.18-85.78 years), and 48.2% were men. The manual grading system detected RDR in 399 (41.48%) participants. The AIDRScreening system had a sensitivity of 86.72% (95% CI: 83.39-90.05%) and a specificity of 96.09% (95% CI: 94.14-97.54%) in the detection of RDR, and a false-positive rate of 3.91%. The diagnostic accuracy gain rate of the AIDRScreening system was 16.57% higher than that of the investigator, while the average diagnostic time gain rate was -37.32% lower. Conclusions: The automated AIDRScreening system can detect RDR with high accuracy, but cannot detect maculopathy. The implementation of the AIDRScreening system may increase the efficiency of DR screening.