Natural Image Reconstruction from fMRI Based on Node-Edge Interaction and Multi-Scale Constraint.

Reconstructing natural stimulus images using functional magnetic resonance imaging (fMRI) is one of the most challenging problems in brain decoding and is also the crucial component of a brain-computer interface. Previous methods cannot fully exploit the information about interactions among brain regions. In this paper, we propose a natural image reconstruction method based on node-edge interaction and a multi-scale constraint. Inspired by the extensive information interactions in the brain, a novel graph neural network block with node-edge interaction (NEI-GNN block) is presented, which can adequately model the information exchange between brain areas via alternatively updating the nodes and edges. Additionally, to enhance the quality of reconstructed images in terms of both global structure and local detail, we employ a multi-stage reconstruction network that restricts the reconstructed images in a coarse-to-fine manner across multiple scales. Qualitative experiments on the generic object decoding (GOD) dataset demonstrate that the reconstructed images contain accurate structural information and rich texture details. Furthermore, the proposed method surpasses the existing state-of-the-art methods in terms of accuracy in the commonly used n-way evaluation. Our approach achieves 82.00%, 59.40%, 45.20% in n-way mean squared error (MSE) evaluation and 83.50%, 61.80%, 46.00% in n-way structural similarity index measure (SSIM) evaluation, respectively. Our experiments reveal the importance of information interaction among brain areas and also demonstrate the potential for developing visual-decoding brain-computer interfaces.

Expression of the SARS-CoV-2 Receptor ACE2 and Protease TMPRSS2 in Ocular Hypertension Eyes of Nonhuman Primate and Human.

PURPOSE: Coronavirus disease 2019 is a disease caused by the novel severe acute respiratory syndrome coronavirus 2. The double-positive of angiotensin-converting enzyme 2 and transmembrane protease serine type 2 have a higher risk of being infected by severe acute respiratory syndrome coronavirus 2. The susceptibility of coronavirus disease 2019 in patients with chronic diseases, especially in different tissues of ocular hypertension eyes like glaucoma, is not yet known. METHODS: An ocular hypertension model was established by laser photocoagulation in rhesus monkeys. The expression of angiotensin-converting enzyme 2 and transmembrane protease serine type 2 in three ocular hypertension eyes and the three control eyes were analyzed using immunofluorescence. RESULTS: No difference was observed between ocular hypertension and control eyes in the expression of angiotensin-converting enzyme 2 and transmembrane protease serine type 2 in the conjunctival epithelium, corneal epithelium, and ciliary muscle. In ocular hypertension eyes and control eyes, angiotensin-converting enzyme 2 and transmembrane protease serine type 2 expression were both observed in the retina. Angiotensin-converting enzyme 2 staining of retinal ganglion cells was found to be significantly higher in ocular hypertension eyes than in control eyes. However, there was no difference in angiotensin-converting enzyme 2 and transmembrane protease serine type 2 expression in retinal vessels and choroidal vessels between ocular hypertension and control eyes. In our study, the expression and distribution of angiotensin-converting enzyme 2 and TMPREE2 in human retina were similar to that of non-human primates as expected. CONCLUSION: Our study confirmed that angiotensin-converting enzyme 2 and transmembrane protease serine type 2 were expressed widely in rhesus monkey eyes. When compared with controls eyes, the expression of angiotensin-converting enzyme 2 was higher in the retinal ganglion cells in ocular hypertension eyes, suggesting that high ocular pressure may affect the patients' ocular susceptibility to severe acute respiratory syndrome coronavirus 2 infection.

Binocular head-mounted chromatic pupillometry can detect structural and functional loss in glaucoma.

Aim: The aim of this study is to evaluate the utility of binocular chromatic pupillometry in detecting impaired pupillary light response (PLR) in patients with primary open-angle glaucoma (POAG) and to assess the feasibility of using binocular chromatic pupillometer in opportunistic POAG diagnosis in community-based or telemedicine-based services. Methods: In this prospective, cross-sectional study, 74 patients with POAG and 23 healthy controls were enrolled. All participants underwent comprehensive ophthalmologic examinations including optical coherence tomography (OCT) and standard automated perimetry (SAP). The PLR tests included sequential tests of full-field chromatic stimuli weighted by rods, intrinsically photosensitive retinal ganglion cells (ipRGCs), and cones (Experiment 1), as well as alternating chromatic light flash-induced relative afferent pupillary defect (RAPD) test (Experiment 2). In Experiment 1, the constricting amplitude, velocity, and time to maximum constriction/dilation were calculated in three cell type-weighted responses, and the post-illumination response of ipRGC-weighted response was evaluated. In Experiment 2, infrared pupillary asymmetry (IPA) amplitude and anisocoria duration induced by intermittent blue or red light flashes were calculated. Results: In Experiment 1, the PLR of POAG patients was significantly reduced in all conditions, reflecting the defect in photoreception through rods, cones, and ipRGCs. The variable with the highest area under the receiver operating characteristic curve (AUC) was time to max dilation under ipRGC-weighted stimulus, followed by the constriction amplitude under cone-weighted stimulus and the constriction amplitude response to ipRGC-weighted stimuli. The impaired PLR features were associated with greater visual field loss, thinner retinal nerve fiber layer (RNFL) thickness, and cupping of the optic disk. In Experiment 2, IPA and anisocoria duration induced by intermittent blue or red light flashes were significantly greater in participants with POAG than in controls. IPA and anisocoria duration had good diagnostic value, correlating with the inter-eye asymmetry of visual field loss. Conclusion: We demonstrate that binocular chromatic pupillometry could potentially serve as an objective clinical tool for opportunistic glaucoma diagnosis in community-based or telemedicine-based services. Binocular chromatic pupillometry allows an accurate, objective, and rapid assessment of retinal structural impairment and functional loss in glaucomatous eyes of different severity levels.

Evaluation of the Glaucomatous Macular Damage by Chromatic Pupillometry.

INTRODUCTION: This study aimed to examine the performance of binocular chromatic pupillometry for the objective and rapid detection of primary open-angle glaucoma (POAG), and to explore the association between pupillary light response (PLR) features and structural glaucomatous macular damage. METHODS: Forty-six patients (mean age 41.00 ± 13.03 years) with POAG and 23 healthy controls (mean age 42.00 ± 11.08 years) were enrolled. All participants underwent sequenced PLR tests of full-field, superior/inferior quadrant-field chromatic stimuli using a binocular head-mounted pupillometer. The constricting amplitude, velocity, and time to max constriction/dilation, and the post-illumination pupil response (PIPR) were analyzed. The inner retina thickness and volume measurements were determined by spectral domain optical coherence tomography. RESULTS: In the full-field stimulus experiment, time to pupil dilation was inversely correlated with perifoveal thickness (r = - 0.429, P < 0.001) and perifoveal volume (r = - 0.364, P < 0.001). Dilation time (AUC 0.833) showed good diagnostic performance, followed by the constriction amplitude (AUC 0.681) and PIPR (AUC 0.620). In the superior quadrant-field stimulus experiment, time of pupil dilation negatively correlated with inferior perifoveal thickness (r = - 0.451, P < 0.001) and inferior perifoveal volume (r = - 0.417, P < 0.001). The dilation time in response to the superior quadrant-field stimulus showed the best diagnostic performance (AUC 0.909). In the inferior quadrant-field stimulus experiment, time to pupil dilation (P < 0.001) correlated well with superior perifoveal thickness (r = - 0.299, P < 0.001) and superior perifoveal volume (r = - 0.304, P < 0.001). CONCLUSION: The use of chromatic pupillometry offers a patient-friendly and objective approach to detect POAG, while the impairment of PLR features may serve as a potential indicator of structural macular damage.

Visual Crowding Reveals Field- and Axis-Specific Cortical Miswiring After Long-Term Axial Misalignment in Strabismic Patients Without Amblyopia.

Purpose: Inspired by physiological and neuroimaging findings that revealed squint-induced modification of cortical volume and visual receptive field in early visual areas, we hypothesized that strabismic eyes without amblyopia manifest an increase in critical spacing of visual crowding, an essential bottleneck on object recognition and reliable psychophysical index of cortical organization. Methods: We used real-time eye tracking to ensure gaze-contingent display and examined visual crowding in patients with horizontal concomitant strabismus (both esotropia and exotropia) but without amblyopia and age-matched normal controls. Results: Nineteen patients with exotropia (12 men, mean ± SD = 22.89 ± 7.82 years), 21 patients with esotropia (10 men, mean ± SD = 23.48 ± 6.95 years), and 14 age-matched normal controls (7 men, mean ± SD = 23.07 ± 1.07 years) participated in this study. We found that patients with strabismus without amblyopia showed significantly larger critical spacing with nasotemporal asymmetry in only the radial axis that related to the strabismus pattern, with exotropia exhibiting stronger temporal hemifield crowding and esotropia exhibiting stronger nasal hemifield crowding, in both the deviated and fixating eyes. Moreover, the magnitude of crowding change was related to the duration and degree of strabismic deviation. Conclusions: Using visual crowding as a psychophysical index of cortical organization, our study demonstrated significantly greater peripheral visual crowding with nasotemporal asymmetry in only the radial axis in patients with strabismus without amblyopia, indicating the existence of hemifield- and axis-specific miswiring of cortical processing in object recognition induced by long-term adaptation to ocular misalignment.

Use of a tissue clearing technique combined with retrograde trans-synaptic viral tracing to evaluate changes in mouse retinorecipient brain regions following optic nerve crush.

Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve regeneration. However, morphological assessments of retinorecipient regions are limited by the opacity of brain tissue. In this study, we used an innovative tissue cleaning technique combined with retrograde trans-synaptic viral tracing to observe changes in retinorecipient regions connected to retinal ganglion cells in mice after optic nerve injury. Specifically, we performed light-sheet imaging of whole brain tissue after a clearing process. We found that pseudorabies virus 724 (PRV724) mostly infected retinal ganglion cells, and that we could use it to retrogradely trace the retinorecipient regions in whole tissue-cleared brains. Unexpectedly, PRV724-traced neurons were more widely distributed compared with data from previous studies. We found that optic nerve injury could selectively modify projections from retinal ganglion cells in the hypothalamic paraventricular nucleus, intergeniculate leaflet, ventral lateral geniculate nucleus, central amygdala, basolateral amygdala, Edinger-Westphal nucleus, and oculomotor nucleus, but not the superior vestibular nucleus, red nucleus, locus coeruleus, gigantocellular reticular nucleus, or facial nerve nucleus. Our findings demonstrate that the tissue clearing technique, combined with retrograde trans-synaptic viral tracing, can be used to objectively and comprehensively evaluate changes in mouse retinorecipient regions that receive projections from retinal ganglion cells after optic nerve injury. Thus, our approach may be useful for future estimations of optic nerve injury and regeneration.

The risk of glaucoma associated with phacomatosis cesioflammea and phacomatosis cesioflammeo-marmorata

Background: The ocular features of phacomatosis pigmentovascularis (PPV) have rarely been reported, and glaucoma is the leading cause of blindness in patients with this condition. To protect vision in these patients, it is important to identify glaucoma as early as possible. Objectives: To systematically report the systemic and ocular manifestations of phacomatosis cesioflammea and phacomatosis cesioflammeo-marmorata, and to investigate a glaucoma risk scoring system. Materials & Methods: In this prospective study, patients with PPV from 2014 to 2021 were included. Clinical information was collected, and associations with glaucoma were evaluated. The suitability of the scoring system was assessed. A systematic literature review and analysis of reported cases of PPV was performed. Results: A total of 28 participants with PPV were included. Their ocular findings were similar, ranging from episcleral hyperpigmentation (78.5%), glaucoma (75%), choroid haemangioma (38%), and retinal vascular abnormalities (48%), to hyperpigmentation of the cornea, iris, lens and fundus. Glaucoma was associated with multiple factors, especially a thick choroid (odds ratio: 2.61; p = 0.008) and a diffuse mass-type of episcleral hyperpigmentation (odds ratio: 41.3; p = 0.027). The risk scoring system was characterized by high sensitivity (84%) and specificity (80%; AUC = 0.91) in predicting glaucoma. Conclusion: In addition to involving the systemic system, phacomatosis cesioflammea and phacomatosis cesioflammeo-marmorata also represent a specific spectrum of ophthalmic vascular malformations and hyperpigmentation. Early and periodic detailed ocular examination are recommended. The novel scoring system will help to tailor follow-up for visual protection.

Protocol for laser-induced chronic ocular hypertension and intracameral injection in nonhuman primates.

Laser-induced hypertension in nonhuman primates is used to mimic human glaucoma, the leading cause of irreversible blindness. In this protocol, we detail steps for laser-induced ocular hypertension in nonhuman primates by laser photocoagulation of the trabecular meshwork and subsequent intracameral injection. We further describe recording and evaluation of intraocular pressure changes and peripapillary retinal nerve fiber layer thickness. This protocol can assist researchers improve the success rate and repeatability of the procedure and reduce the number of nonhuman primates needed. For complete details on the use and execution of this protocol, please refer to Sun et al. (2022).

Erratum: Long-term and potent IOP-lowering effect of IκBα-siRNA in a nonhuman primate model of chronic ocular hypertension.

[This corrects the article DOI: 10.1016/j.isci.2022.104149.].

Long-term and potent IOP-lowering effect of IκBα-siRNA in a nonhuman primate model of chronic ocular hypertension.

Glaucoma is one of the most common causes of irreversible blindness. It is acknowledged that lowering intraocular pressure (IOP) is the effective treatment to slow glaucoma disease progression. The main obstacle of existing drugs is that the effect of reducing IOP does not last long. Degradation of IκB stimulates the transcription of NF-κB, which could upregulate the expression of matrix metalloproteinases (MMPs). Whether a IκB-targeted gene therapy works in glaucoma is unclear. Here, we established a chronic ocular hypertension (COHT) model in rhesus monkey by laser photocoagulation and verified that intracameral delivery of IκBα-siRNA showed long-lasting and potent effects of reducing IOP without obvious inflammation in monkeys with COHT. We also verified that IκBα-siRNA could increase the expressions of MMP2 and MMP9 by knocking down IκBα in vitro and in vivo. Our results in nonhuman primates indicated that IκBα-siRNA may become a promising therapeutic approach for the treatment of glaucoma.

Radiation optic neuropathy diagnosis in nasopharyngeal carcinoma patients with radiation encephalopathy.

AIM: To develop a suitable radiation optic neuropathy (RON) diagnostic model based on optical coherence tomography angiography (OCTA) and to reveal the correlation between endocrine and OCTA indexes in nasopharyngeal carcinoma (NPC) patients with radiation encephalopathy (RE). METHODS: This retrospective cross-sectional study included seventy-seven male NPC patients with RE following radiotherapy (41 non-RON and 36 RON). Endocrine and OCTA indexes were collected. The macular and peripapillary vessel density (VD) were automatically analyzed using AngioVue 2.0 of the RTVue XR Avanti device. The OCTA indexes were included in the multivariable binary logistic regression model between non-RON and RON. For all RE patients, a multiple linear regression was performed between each of the OCTA indexes and the endocrine indexes. RESULTS: Compared to non-RON, enlarged foveal avascular zone (FAZ) area, reduced superficial vascular plexus (SVP) VD and radial peripapillary capillary plexus density, reduced full retinal thickness, ganglion cell complex thickness and peripapillary retinal nerve fiber layer thickness were observed in RON. Foveal VD in the SVP was included in the RON diagnostic model. Free triiodothyronine (FT3) showed a significantly negative correlation with the FAZ area but showed a significantly positive correlation with foveal VD in the SVP and deep vascular plexus, peripapillary VD and thickness and parafoveal and perifoveal thickness. CONCLUSIONS: OCTA may be useful for diagnosing RON in male NPC patients with RE following radiotherapy. Reduced FT3 and foveal VD in SVP may be sensitive in screening RON in these patients.

A Classification Tree Model with Optical Coherence Tomography Angiography Variables to Screen Early-Stage Diabetic Retinopathy in Diabetic Patients.

AIM: To establish a classification tree model in DR screening and to compare the DR screening accuracy between the classification tree model and the logistic regression model in type 2 diabetes mellitus (T2DM) patients based on OCTA variables. METHODS: Two hundred forty-one eyes of 241 T2DM patients were included and divided into two groups: the development cohort and the validation cohort. Optical coherence tomography angiography (OCTA) images were acquired in these patients. The data of foveal avascular zone area, superficial capillary plexus (SCP) density, and deep capillary plexus (DCP) density were exported after automatically analyzing the macular 6 × 6 mm OCTA images, while the data of radial peripapillary capillary plexus (RPCP) density was exported after automatically analyzing the optic nerve head 4.5 × 4.5 mm OCTA images. These OCTA variables were adopted to establish and validate the logistic regression model and the classification tree model. The area under the curve (AUC), sensitivity, specificity, and statistical power for receiver operating characteristic curves of two models were calculated. RESULTS: In the logistic regression model, best-corrected visual acuity (BCVA) (LogMAR) and SCP density were entered (BVCA : OR= 60.30, 95% CI= [2.40, 1513.82], p = 0.013; SCP density: OR= 0.86, 95% CI= [0.78, 0.96], p = 0.006). The AUC, sensitivity, and specificity for detecting early-stage DR (mild to moderate NPDR) in the development cohort were 0.75 (95% CI: [0.66, 0.85]), 63%, and 83%, respectively. The AUC, sensitivity, and specificity in the validation cohort were 0.75 (95% CI: [0.66, 0.84]), 79%, and 72%, respectively. In the classification tree model, BVCA (LogMAR), DM duration, SCP density, and DCP density were entered. The AUC, sensitivity, and specificity for detecting early-stage DR were 0.72 (95% CI: [0.60, 0.84]), 66%, and 76%, respectively. The AUC, sensitivity, and specificity in the validation cohort were 0.74 (95% CI: [0.65, 0.83]), 74%, and 72%, respectively. The statistical power of the development and validation cohorts in two models was all more than 99%. CONCLUSIONS: Compared to the logistic regression model, the classification tree model has similar accuracy in predicting early-stage DR. The classification tree model with OCTA variables may be a simple tool for clinical practitioners to identify early-stage DR in T2DM patients. Moreover, SCP density is significantly reduced in mild-to-moderate NPDR eyes and might be a biomarker in early-stage DR detection. Further improvement and validation of the DR diagnostic model are awaiting to be performed.

Relative peripheral refraction characteristics and their relationship with retinal microvasculature in young adults: Using a novel quantitative approach.

AIM: To analyze relative peripheral refraction (RPR) characteristics in young adults and to investigate the relationship between RPR and retinal microvasculature using multispectral refractive tomography (MRT), a novel quantitative approach. METHODS: This cross-sectional study included 278 eyes of 139 young adults. All eyes underwent complete ophthalmic examinations, including MRT, optical coherence tomography angiography (OCTA) and other ocular examinations. Refraction difference values (RDVs) among different rings/sectors were compared using one-way ANOVA in bilateral eyes. Stepwise multiple linear regression analyses were performed between ∆RDV in different rings and ∆density/thickness value in OCTA. RESULTS: Among the different rings, the 30°-45°/45°-53° RDV was significantly greater than the inner rings in both eyes. Among the different sectors, RDV in the superior sector was most significantly reduced among all sectors, and RDV in the nasal sector was significantly greater than that in the temporal sector. In the stepwise multiple linear regression analyses, ∆RDV was negatively correlated with ∆radial peripapillary capillary plexus density and ∆macular thickness. CONCLUSIONS: MRT may be a useful tool in RPR quantitative assessment. RPR and some OCTA indexes might be closely correlated. Further research should be conducted to investigate the relationships among these indexes in young adults.

Network Pharmacology-Based Approach to Comparatively Predict the Active Ingredients and Molecular Targets of Compound Xueshuantong Capsule and Hexuemingmu Tablet in the Treatment of Proliferative Diabetic Retinopathy.

BACKGROUND: Compound Xueshuantong capsule (CXC) and Hexuemingmu tablet (HXMMT) are two important Chinese patent medicines (CPMs) frequently used to treat proliferative diabetic retinopathy (PDR), especially when complicated with vitreous hemorrhage (VH). However, a network pharmacology approach to understand the therapeutic mechanisms of these two CPMs in PDR has not been applied. OBJECTIVE: To identify differences in the active ingredients between CXC and HXMMT and to comparatively predict and further analyze the molecular targets shared by these CPMs and PDR. Materials and methods. The differentially expressed messenger RNAs (mRNAs) between normal retinal tissues in healthy individuals and active fibrovascular membranes in PDR patients were retrieved from the Gene Expression Omnibus database. The active ingredients of CXC and HXMMT and the targets of these ingredients were retrieved from the Traditional Chinese Medicine Systems Pharmacology database. The intersections of the CPM (CXC and HXMMT) targets and PDR targets were determined. Then, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed, and the ingredient-target networks, protein-protein interaction networks, and KEGG-target (KEGG-T) networks were constructed. RESULTS: CXC contains 4 herbs, and HXMMT contains 19. Radix salviae is the only herb common to both. CXC had 34 potential therapeutic targets in PDR, while HXMMT had these 34 and 10 additional targets. Both CPMs shared the following main processes: response to reactive oxygen species and oxidative stress, regulation of blood vessel diameter and size, vasoconstriction, smooth muscle contraction, hemostasis, and blood coagulation. The shared pathways included the AGE-RAGE signaling pathway in diabetic complications, TNF signaling pathway, relaxin signaling pathway, and IL-17 signaling pathway. CONCLUSIONS: Both CXC and HXMMT include components effective at treating PDR and affect the following main processes: response to reactive oxygen species and oxidative stress, regulation of blood vessels, and blood coagulation. Radix salviae, the only herb common to both CPMs, contains many useful active ingredients. The PDR-CXC and PDR-HXMMT networks shared 34 common genes (RELA, HSPA8, HSP90AA, HSP90AB1, BRCA, EWSR1, CUL7, HNRNPU, MYC, CTNNB1, MDM2, YWHAZ, CDK2, AR, FN1, HUWE1, TP53, TUBB, EP300, GRB2, VCP, MCM2, EEF1A1, NTRK1, TRAF6, EGFR, PRKDC, SRC, HDAC5, APP, ESR1, AKT1, UBC, and COPS5), and the PDR-HXMMT network has 10 additional genes (RNF2, VNL, RPS27, COPS5, XPO1, PARP1, RACK1, YWHAB, and ITGA4). The top 5 pathways with the highest gene ratio in both networks were the AGE-RAGE signaling pathway in diabetic complications, TNF signaling pathway, relaxin signaling pathway, IL-17 signaling pathway, and focal adhesion. Additional pathways such as neuroactive ligand-receptor interaction, chemokine signaling pathway, and AMPK signaling pathway were enriched with HXMMT targets. Thus, HXMMT has more therapeutic targets shared by different active ingredients and more abundant gene functions than CXC, which may be two major reasons why HXMMT is more strongly recommended than CXC as an auxiliary treatment for new-onset VH secondary to PDR. However, the underlying mechanisms still need to be further explored.

Reduced Dendritic Spines in the Visual Cortex Contralateral to the Optic Nerve Crush Eye in Adult Mice.

Purpose: To determine alteration of dendritic spines and associated changes in the primary visual cortex (V1 region) related to unilateral optic nerve crush (ONC) in adult mice. Methods: Adult unilateral ONC mice were established. Retinal nerve fiber layer (RNFL) thickness was measured by spectral-domain optical coherence tomography. Visual function was estimated by flash visual evoked potentials (FVEPs). Dendritic spines were observed in the V1 region contralateral to the ONC eye by two-photon imaging in vivo. The neurons, reactive astrocytes, oligodendrocytes, and activated microglia were assessed by NeuN, glial fibrillary acidic protein, CNPase, and CD68 in immunohistochemistry, respectively. Tropomyosin receptor kinase B (TrkB) and the markers in TrkB trafficking were estimated using western blotting and co-immunoprecipitation. Transmission electron microscopy and western blotting were used to evaluate autophagy. Results: The amplitude and latency of FVEPs were decreased and delayed at 3 days, 1 week, 2 weeks, and 4 weeks after ONC, and RNFL thickness was decreased at 2 and 4 weeks after ONC. Dendritic spines were reduced in the V1 region contralateral to the ONC eye at 2, 3, and 4 weeks after ONC, with an unchanged number of neurons. Reactive astrocyte staining was increased at 2 and 4 weeks after ONC, but oligodendrocyte and activated microglia staining remained unchanged. TrkB was reduced with changes in the major trafficking proteins, and enhanced autophagy was observed in the V1 region contralateral to the ONC eye. Conclusions: Dendritic spines were reduced in the V1 region contralateral to the ONC eye in adult mice. Reactive astrocytes and decreased TrkB may be associated with the reduced dendritic spines.

SP1-mediated upregulation of LINGO-1 promotes degeneration of retinal ganglion cells in optic nerve injury.

BACKGROUNDS: Insults to the axons in the optic nerve head are the primary cause of loss of retinal ganglion cells (RGCs) in traumatic, ischemic nerve injury or degenerative ocular diseases. The central nervous system-specific leucine-rich repeat protein, LINGO-1, negatively regulates axon regeneration and neuronal survival after injury. However, the upstream molecular mechanisms that regulate LINGO-1 signaling and contribute to LINGO-1-mediated death of RGCs are unclear. METHODS: The expression of SP1 was profiled in optic nerve crush (ONC)-injured RGCs. LINGO-1 level was examined after SP1 overexpression by qRT-PCR. Luciferase assay was used to examine the binding of SP1 to the promoter regions of LINGO-1. Primary RGCs from rat retina were isolated by immunopanning and RGCs apoptosis were determined by Tunnel. SP1 and LINGO-1 expression was investigated using immunohistochemistry and Western bolting. Neuroprotection was assessed by RGC counts, RNFL thickness, and VEP tests after inhibition of SP1 shRNA. RESULTS: We demonstrate that SP1 was upregulated in ONC-injured RGCs. SP1 was bound to the LINGO-1 promoter, which led to increased expression of LINGO-1. Treatment with recombinant Nogo-66 or LINGO-1 promoted apoptosis of RGCs cultured under serum-deprivation conditions, while silencing of SP1 promoted the survival of RGCs. SP1 and LINGO-1 colocalized and were upregulated in ONC-injured retinas. Silencing of SP1 in vivo reduced LINGO-1 expression and protected the structure of RGCs from ONC-induced injury, but there was no sign of recovery in VEP. CONCLUSIONS: Our findings imply that SP1 regulates LINGO-1 expression in RGCs in the injured retina and provide insight into mechanisms underlying LINGO-1-mediated RGC death in optic nerve injury.

Radiation-Induced Optical Coherence Tomography Angiography Retinal Alterations in Patients With Nasopharyngeal Carcinoma.

Aim: The aim of the study was to investigate the early neurovascular alterations of the retina in radiation encephalopathy (RE) patients with normal-ranged visual acuity after radiotherapy for nasopharyngeal carcinoma. Methods: Fifty-five RE patients and 54 healthy age-matched subjects were enrolled in this retrospective cross-sectional case-control study. The best corrected visual acuity (LogMAR) of the included eye should not be more than 0. The vessel density and thickness of different locations in the retina were acquired automatically using optical coherence tomography angiography (OCTA). The data were then compared between the RE patients and the controls. The location included the whole retina, the superficial vascular plexus (SVP)/the ganglion cell complex (GCC), the deep vascular plexus (DVP), and the choroid in the macular area, as well as the inside disc and peripapillary area in the optic nerve head (ONH). The risk factors in OCTA retinal impairments were analyzed using a backward multiple linear regression. The relationships between mean deviation (MD) and pattern standard deviation (PSD) in the visual field (VF) and the OCTA parameters were also analyzed in RE patients. Results: The vessel density of the GCC was significantly reduced in RE patients compared with controls (p = 0.018), and the reductions were mainly shown in the parafoveal (p = 0.049) and perifoveal fields (p = 0.006). The thickness of the GCC was correspondingly reduced (whole image GCC mean thickness: p = 0.044; parafoveal thickness: p = 0.038; perifoveal thickness: p = 0.038). In addition, the sub-foveal choroidal thickness (p = 0.039) was also reduced in RE patients. The vessel density of the GCC (R 2 = 0.643) and DVP (R 2 = 0.777) had a significant positive correlation with high-density lipoprotein cholesterol (HDL-C) and apolipoprotein A1 (ApoA1) and had a significant negative correlation with age (GCC: HDL-C, ß = 29.89, p = 0.005; ApoA1, ß = 78.92, p = 0.002; age, ß = -0.886, p = 0.001; DVP: HDL-C, ß = 40.09, p = 0.003; ApoA1, ß = 62.65, p = 0.013; age, ß = -1.31, p = 0.001). The vessel density of the GCC also had a significant negative correlation with apolipoprotein B (ApoB) (ß = -32.18, p = 0.006). In the VF, MD had a significant positive correlation with the vessel density inside disc (R 2 = 0.241, ß = 0.304, p = 0.045), whereas PSD showed a significant negative correlation with the vessel density inside disc and the average GCC thickness, respectively (R 2 = 0.437; vessel density inside disc, ß = -0.358, p = 0.004; average GCC thickness, ß = -0.510, p < 0.001). Conclusion: With the aid of OCTA, we found that neurovascular alterations of the retina may exist in RE patients with normal-ranged visual acuity. Herein, we suggest the implementation of OCTA to assist ophthalmologists in the early detection and consistent monitoring of radiation-related eye diseases to avoid delayed diagnosis.

Inhibition of the leucine-rich repeat protein lingo-1 enhances RGC survival in optic nerve injury.

Leucine-rich repeat and immunoglobulin-like domain-containing nogo receptor-interacting protein 1 (lingo-1) is selectively expressed on neurons and oligodendrocytes in the central nervous system and acts as a negative regulator in neural repair, implying a potential role in optic neuropathy. The aim of the present study was to determine whether adeno-associated virus serotype 2 (AAV2) vector-mediated transfer of lingo-1 short hairpin RNA could reduce nerve crush-induced axonal degeneration and enhance axonal regeneration following optic nerve (ON) injury in vivo. The expression of lingo-1 was knocked down in vivo using a green fluorescent protein (GFP)-tagged AAV2 encoding lingo-1 shRNA via intravitreal injection in adult Sprague-Dawley rats. Silencing effects of AAV2-lingo-1-shRNA were confirmed by detecting GFP labelling of RGCs, and by quantifying lingo-1 expression levels with reverse transcription-quantitative polymerase chain reaction and western blotting. Rats received an intravitreal injection of AAV2-lingo-1-shRNA or negative control shRNA. The ON crush (ONC) injury was performed 2 weeks after the intravitreal injection. RGC density, lesion volume of the injured ON and the visual electrophysiology [flash visual evoked potential (F-VEP)] at different time points post-injury were determined. Transduction with lingo-1-shRNA decreased lingo-1 expression levels and promoted RGC survival following ONC. Lingo-1-shRNA promoted ON tissue repair and functional recovery. The mechanism underlying the effect of AAV2-lingo-1-shRNA on RGCs may be the phosphorylation of protein kinase B (Akt) at Ser473 and activation of the Akt signaling pathway acting downstream of lingo-1. The results of the current study indicate that the inhibition of lingo-1 may enhance RGC survival and facilitate functional recovery following ON injury, representing a promising potential strategy for the repair of optic neuropathy.

Comparison between non-visualized polyps and visualized polyps on optical coherence tomography angiography in polypoidal choroidal vasculopathy.

PURPOSE: To determine the underlying reasons for the non-visualization of polyps on en face optical coherence tomography angiography (OCTA) in patients with polypoidal choroidal vasculopathy (PCV). METHODS: A cross-sectional study of consecutive treatment-naïve 30 eyes with active PCV was included. Results of fundus photography, fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA), spectral domain optical coherence tomography (SD-OCT), and en face OCTA were analyzed. RESULTS: A total of 64 active polyps were found on FFA and ICGA in 30 eyes. On OCTA, 42/64 (65.6%) polyps were visualized, while 22/64 (34.4%) polyps were non-visualized. There were no significant differences in the size (P = 0.723) and filling time of polyps (P = 0.558) between the two groups. However, polypoidal lesions were less common in the non-visualized group (P < 0.001). The height of the polyps on SD-OCT was 243.95 ± 114.24 µm in the non-visualized group, which was higher than those (188.00 ± 87.93 µm) in the visualized group (P = 0.048). Moreover, more pulsatile polyps (72.7%) were found in the non-visualized group than those (2.4%) in the visualized group (P < 0.001). Four of the 22 polyps in the non-visualized group (18.2%) were located under a thick subretinal hemorrhage, and two of 22 invisible polyps (9.6%) located under and parallel to the retinal vessel in the inner layer of retina. CONCLUSIONS: Our results revealed that the height of the polyps, and not the size and pulsation of the polyps, correlated with the visualization of the polyps on OCTA. Polyps that were pulsating in early ICGA were difficult to be visualized on OCTA, which is the most possible reason for the non-visualization. Coverage with thick subretinal hemorrhage or retina vessels was another reason for the non-visualization of the polyps in active PCV on OCTA.

Mechanism of fibrosis inhibition in laser induced choroidal neovascularization by doxycycline.

To explore the mechanisms underlying doxycycline suppression of fibrosis in laser-induced choroidal neovascularization (LCNV), C57BL/6J male mice (aged from 6 to 8 weeks) received intraperitoneal injections of PBS/doxycycline solution from one day before laser injury until they were sacrificed. Leakage was assessed by FA, and CNV (stained by IB4) or fibrosis (stained by collagen type I) size was measured. The percentage of Pan-keratin+α-SMA+ cells was counted in the eyes' cryostat sections by immunohistochemistry. qPCR was used to measure the mRNA of markers of pan-macrophages, M1 and M2-type macrophages (M1 and M2), markers of EMT, and markers in the downstream of STAT6 signaling. Western blotting was used to analyze the expression of Arg-1, α-SMA, E-cadherin, pSTAT6 and STAT6. Our data showed that doxycycline inhibited leakage from CNV, areas of CNV on day 7 and day 14, and suppressed fibrosis, and the ratio of fibrotic/angiogenic areas during day 7 to day 35. We also showed attenuation of EMT in the doxycycline group. The percentage of Pan-keratin+α-SMA+ cells was lower in the doxycycline group than in the control group. The mRNA and protein levels of mesenchymal markers were downregulated in the doxycycline group, while the epithelial marker was upregulated. In addition, our data showed that the protein expression of Arg-1, the mRNA expression of M1 and M2-markers, were both inhibited by doxycycline, while the level of pan-macrophages (f4/80) showed no significant difference in two groups. Finally, our results showed that doxycycline was able to modulate the STAT6 signaling in transcript and protein levels. Accordingly, we suggested that the mechanism of doxycycline-mediated inhibition of fibrosis in CNV occurs through the STAT6 pathway.