Investigation of endothelial cell pathological characteristics in murine choroidal neovascularization model based on single-cell RNA sequencing / 中华实验眼科杂志

Objective:To investigate the molecular expression and pathological features of endothelial cell (EC) in a murine model of choroidal neovascularization (CNV) based on single-cell RNA sequencing (scRNA-seq).Methods:Six C57BL/6 mice aged 6-8 weeks were randomly divided into two groups, with 3 mice in each group.Bilateral eyeballs were enucleated.The choroidal tissues from the two groups were isolated by shearing the complex and scraping the choroid, respectively.Single-cell suspension was prepared by continuous digestion with trypsin/type Ⅰ collagenase at 37 ℃, and the cell viability and EC ratio were detected by flow cytometry to determine the preparation method of single-cell suspension.Another 6 mice were randomly assigned into the control group and the CNV group, with 3 mice in each group.The CNV model was induced by laser photocoagulation and single-cell suspensions were prepared 7 days after modeling.Gene expression library construction was performed using the Chromi-um (10x Genomics) instrument.High throughput sequencing was performed using the Illumina Novaseq6000 to obtain the expression matrix.The EC subpopulations were classified according to previous researches and the Cellmarker database.Pseudo-time analysis was performed in EC, revealing the gene expression matrix of different states.CNV-EC were further selected with preliminary analysis of the expression characteristics.Another 6 mice were selected to establish the CNV model and eyeball frozen sections were prepared 7 days after modeling.Expression and distribution as well as the area percentage of EC marker Pecam1, mitochondrial outer membrane proteins Tomm20 and mt-Co1, and capillary markers Kdr and Plvap were observed by immunofluorescence staining, and the vascular diameter was calculated.The use and care of animals followed the ARVO statement.This study protocol was approved by the Experimental Animal Welfare and Ethics Committee of Air Force Military Medical University (No.20200181).Results:The cell viability of the single-cell suspension prepared from choroidal-scleral fragments and choroidal scrapings was 99.4% and 99.1%, respectively, both of which met the sequencing requirements.The percentage of EC detected by flow cytometry was approximately 1.58%.The scRNA-seq result revealed that both the normal control and CNV groups contained 13 choroidal cell clusters.Compared with the normal control group, the proportions of rod/cone photoreceptor cells, EC and hematopoietic cells all increased, while the retinal pigment epithelium (RPE) and Schwan cells reduced in the CNV group.Among all clusters, EC constituted 18.4%.The pseudo-time analysis demonstrated that EC could be further divided into 4 states.The percentage of state 2 EC was 29.1% in the CNV group, which was significantly higher than 9.5% in the normal control group.Differentially expressed gene analysis showed that the expression of mitochondrion-related genes, including mt-Nd4 and mt-Atp6, were upregulated in state 2 EC, while capillary-related genes, including Kdr and Esm1, were downregulated.Immunofluorescent staining revealed that the area of Tomm20 and mt-Co1 in Pecam1-positive EC in the CNV area was (19.50±4.68)% and (4.64±2.82)%, respectively, which were both higher than (3.00±2.09)% and (0.18±0.34)% in normal area ( t=7.88, 3.84; both at P<0.01). The area of Kdr and Plvap in Pecam1-positive EC in the CNV area was (1.50±0.29)% and (0.79±0.97)%, respectively, which were both lower than (31.30±5.44)% and (10.43±2.28)% in the normal area ( t=13.40, 9.48; both at P<0.01). The vascular diameter in the CNV area was (5.52±1.85)μm, which was larger than (4.21±1.84)μm in the normal area ( t=9.57, P<0.001). Conclusions:When CNV occurs, the proportion of EC in choroid increases, and CNV-EC shows pathologic features of mitochondrial metabolic activation and loss of capillary properties, suggesting the mitochondrial activation of EC may play a role in the formation of CNV.

Mechanism of curcumin inhibiting choroidal neovascularization in brown Norway rat / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To investigate the mechanism of curcumin inhibiting the choroidal neovascularization(CNV)of brown Norway(BN)rats.METHODS: CNV model of 36 BN rats was established through laser photocoagulation induction, and they were divided into 6 groups with 6 rats in each group. Normal group was fed normally with no intervention, while 532nm laser photocoagulation was used to establish a experimental CNV model in BN rats. Rats after modeling were respectively intervened for 14d and divided into model group, ranibizumab group, curcumin low [100mg/(kg·d)], medium [200mg/(kg·d)], and high [400mg/(kg·d)] dose group. The model group was given intragastric administration of saline for 14d, ranibizumab(10mg/mL, 0.2mL/dose)was injected at 2d after photocoagulation with 5μL once for rats in ranibizumab group, and different concentrations of curcumin were intragastrically administrated to the rats in low, medium and high groups for 14d. Fundus photography, fundus fluorescein angiography(FFA)and indocyanine green angiography(ICGA)examination were performed at 14d after photocoagulation. Ocular histopathological specimens of rats with CNV were made, and the central thickness of CNV were observed by HE staining. Ocular histopathological specimens were made, and the expressions of AKT/p-AKT/HIF-1α/VEGF signaling pathway-related proteins were observed by immunohistochemistry. The mRNA relative expressions of AKT/HIF-1α/VEGF factor in CNV tissues were detected by RT-qPCR, and the protein expressions of AKT/p-AKT/HIF-1α/VEGF factor in CNV tissues were detected by Western-blot.RESULTS: CNV generation rates in the model group, the ranibizumab group, and the low, medium and high-dose curcumin groups were 78.18%, 73.21%, 77.19%, 75.86%, 74.55%, respectively, which were higher than 70%. The average absorbance were 182.12±6.59, 119.22±8.03, 166.45±8.33, 164.34±5.69, 149.22±6.45, respectively; the ranibizumab group was significantly lower than the model group(P&#x0026;#x003C;0.05); the low-dose, medium-dose and high-dose groups were significantly higher than the ranibizumab group(P&#x0026;#x003C;0.05), and the curcumin high-dose group was significantly lower than the model group(P&#x0026;#x003C;0.05). HE staining showed that the retinal tissue structure of BN rats in normal group was clear and neatly arranged. The central thickness of CNV in the ranibizumab group was significantly reduced at 14d after photocoagulation compared with the model group(P&#x0026;#x003C;0.05); While the curcumin high-dose group was significantly reduced compared with the model group(P&#x0026;#x003C;0.05), but increased when compared with ranibizumab group(P&#x0026;#x003C;0.05). Immunohistochemistry results showed that AKT, p-AKT, HIF-1α, and VEGF factors were negatively expressed in the retinal tissue structure of BN rats in the normal group, and no brown-yellow reactants were found. The expression of AKT, p-AKT, HIF-1α, and VEGF factors in the model group were higher than that in the normal group at 14d after photocoagulation(P&#x0026;#x003C;0.05); the ranibizumab group was lower than the model group(P&#x0026;#x003C;0.05). While the expression of the curcumin high-dose group was significantly decreased compared with the model group(P&#x0026;#x003C;0.05), but significantly increased when compared with ranibizumab group(P&#x0026;#x003C;0.05). The mRNA results showed that the relative expression levels of AKT, HIF-1α and VEGF mRNA in the model group at 14d after photocoagulation were higher than those of the normal group(P&#x0026;#x003C;0.05); the ranibizumab group was lower than the model group(P&#x0026;#x003C;0.05). While curcumin high-dose group was significantly decreased compared with the model group(P&#x0026;#x003C;0.05), but significantly increased when compared with ranibizumab group(P&#x0026;#x003C;0.05). Western-blot results showed that there was no significant difference in the relative expression of AKT protein among each experimental groups at 14d after photocoagulation. The relative expression of p-AKT protein in the model group was significantly higher than that in the normal group(P&#x0026;#x003C;0.05); the ranibizumab group was significantly lower than the model group(P&#x0026;#x003C;0.05); the curcumin high-dose group was significantly lower than the model group(P&#x0026;#x003C;0.05). The relative expression levels of HIF-1α protein were significantly higher in the model group than in the normal group(P&#x0026;#x003C;0.05), and the ranibizumab group was lower than in the model group(P&#x0026;#x003C;0.05). The relative expression levels of HIF-1α protein was lower in the curcumin high-dose group than in the model group(P&#x0026;#x003C;0.05)but higher than ranibizumab group(P&#x0026;#x003C;0.05). The relative expression level of VEGF protein was significantly lower in the curcumin medium/high-dose group than in the model group(P&#x0026;#x003C;0.05).CONCLUSION: Curcumin at 400mg/(kg·d)has an inhibitory effect on CNV in BN rats. The mechanism may be closely related to inhibiting the activation of AKT/p-AKT/HIF-1α/VEGF signaling pathways.

Methyltransferase-like 3-mediated N6-methyladenosine methylation modification regulates the biological activity of vascular endothelial cells via the Notch pathway / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the role and mechanism of methyltransferase-like 3(METTL3)-mediated N6-methyladenosine(m6A)methylation modification in regulating biological activity of vascular endothelial cells in the pathogenesis of choroidal neovascularization.METHODS: Human umbilical vein endothelial cells(HUVEC)cultured in vitro were divided into the following groups: control group(normal culture), low density lipoprotein(LDL)group, fluorescence-labelled LDL(Dil-LDL)group, 12.5μg/mL and 25μg/mL oxidized LDL(ox-LDL)groups, 12.5μg/mL and 25μg/mL fluorescence-labelled ox-LDL(Dil-ox-LDL)groups, DMSO group, STM2457(METTL3 inhibitor)group, DAPT group; and monkey retina-choroidal endothelial cells(RF/6A)cultured in vitro were divided into control group, DMSO group, 12.5 μg/mL ox-LDL group, and DAPT group. Endocytosed lipoprotein level was examined through fluorescence microscopy. RNA m6A methylation level was detected through a dot blot assay. Protein and RNA levels of METTL3 or angiogenesis-related markers were measured through Western blot assays and real-time quantitative polymerase chain reaction(RT-qPCR), respectively. METTL3 expression and localization were investigated through immunofluorescence. Cell migratory and tube formation capacities were assessed through transwell migration and tube formation assays, respectively.RESULTS: Endocytosed lipoprotein levels in HUVECs exposed to Dil-LDL, 12.5μg/mL and 25μg/mL Dil-ox-LDL groups were significantly higher than those in the control group. 12.5μg/mL and 25μg/mL ox-LDL groups significantly increased m6A methylation(all P&#x003C;0.05), METTL3 protein expression(all P&#x003C;0.01), and cell migration and angiogenesis capacities(all P&#x003C;0.01). METTL3 mRNA level was significantly unregulated in the 12.5μg/mL ox-LDL group(P&#x003C;0.05). In comparison to the DMSO group, the addition of STM2457 caused significant decrease in m6A methylation level(P&#x003C;0.05), expression of VEGF and other angiogenesis-related markers(all P&#x003C;0.05), cell migration and angiogenesis capacities(all P&#x003C;0.01)and the expression of NICD(P&#x003C;0.05). However, there were no significant differences in METTL3 protein and mRNA levels(all P&#x003E;0.05). The expression of VEGF and NICD(all P&#x003C;0.05), as well as the ability of cell migration and angiogenesis of RF/6A, was all significantly decreased in the DAPT group compared to the DMSO group(all P&#x003C;0.01).CONCLUSION: METTL3-mediated m6A methylation modification promotes angiogenesis in vascular endothelial cells via the Notch signaling pathway in the pathogenesis of choroidal neovascularization.

Correlation analysis of vision and changes of macular structure and microcirculation in myopic maculopathy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To observe the changes of macular morphology and microcirculation in myopic maculopathy(MM), and investigate theirs correlation and effects on vision.METHODS: Case-control study. A total of 165 patients(189 eyes)with high myopia and 154 healthy volunteers(154 eyes)from October 2016 to December 2018 were selected. According to the classification of Meta-analysis for pathologic myopia(META-PM), participants were divided into M0 group(category 0, 41 eyes), M1 group(category 1, 53 eyes), M2 group(category 2 and 3, 52 eyes), and myopic choroidal neovascularization(mCNV)group(43 eyes). All participants underwent optical coherence tomography angiography(OCTA)examination. Morphological and microcirculation parameters of retina at different layers were compared between groups. Pearson correlation was used to assess the correlation between morphological and microcirculation parameters. Correlations between vision and other parameters were analyzed using multiple linear regression analysis.RESULTS:Foveal full retinal thickness(FRT)and outer retinal thickness(ORT)were all lower in M0, M1 and M2 groups than those of control group(all P&#x003C;0.01). Foveal superficial capillary plexus vessel density(SVD)and deep capillary plexus vessel density(DVD)were all lower in M2 and mCNV groups than those of the control group(all P&#x003C;0.01). Parafoveal FRT and ORT were all lower in M0, M1, M2 and mCNV groups than those of the control group(all P&#x003C;0.01). Parafoveal inner retinal thickness(IRT), SVD and DVD were all lower in M2 and mCNV groups than those of the control group(all P&#x003C;0.01). Subfoveal choroidal thickness(SFCT)and choroid capillaries vessel density(CVD)were all lower in M0, M1, M2 and mCNV groups than those of the control group(all P&#x003C;0.01). Foveal vessel density of retina and choroid were positively correlated with its thickness in patients with MM without CNV(all P&#x003C;0.05). Multivariate analysis showed that axial length(AL), diffuse or patchy chorioretinal atrophy were influencing foctors of best corrected visual acuity(BCVA; all P&#x003C;0.01).CONCLUSION:Retinal morphological changes precede microcirculation changes in MM. Most of all, ORT changes precede IRT changes. Foveal vessel density of retina and choroid were positively correlated with its thickness. The main influencing factors of BCVA were AL and types of MM.

Influencing factors of poor response to anti-vascular endothelial growth factor drug therapy for wet age-related macular degeneration and therapeutic strategy of targeting pericytes / 国际眼科杂志(Guoji Yanke Zazhi)

With the increasing aging population, the incidence of wet age-related macular degeneration(wARMD)is gradually rising. The formation of neovascularization leads to recurrent hemorrhage in the macular region, which is one of the main causes of blindness in the elderly. Currently, the primary clinical treatment for wARMD is intravitreal injection of anti-vascular endothelial growth factor(VEGF)drugs. However, there are still some patients who have poor or no response to anti-VEGF drugs, resulting in suboptimal or ineffective clinical outcomes. Analyzing the specific influencing factors will be beneficial in guiding clinical decision-making. This article reviews the impact of factors such as advanced age, treatment duration, number of injections, characteristics of neovascular lesions, macular structure, intraocular cytokine levels, and genetics on the response to anti-VEGF therapy. In addition, recent studies have found that pericytes, as cellular components of microvascular walls, can influence the sensitivity to anti-VEGF therapy. This review summarizes the current research on the mechanisms of pericytes in poor or non-response to anti-VEGF therapy and discusses targeted strategies focusing on pericytes.

The role of NLRP3/GATA-4/VEGF signal pathway in neovascular age-related macular degeneration / 中国医师杂志

Objective:To explore the potential role of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/GATA-binding protein 4 (GATA-4)/vascular endothelial growth factor (VEGF) signal pathway in neovascular age-related macular degeneration (nAMD).Methods:We applied the TRANSFAC Public database to search the human and mouse VEGF promoters and upstream transcription factors, analyzed the transcription factors that may influence the transcriptional activity of VEGF. The RAW264.7 cells were divided into control group and lipopolysaccharide (LPS) stimulated group (LPS group). Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the activation of NLRP3 inflammasome, and the mRNA levels of GATA-4 and VEGFA. Thus, we applied the specific small molecular NLRP3 inhibitor MCC950 pretreated RAW264.7 cells (LPS+ MCC950 group), and detected the gene expression of NLRP3, Caspase-1, interleukin 1β( IL-1β), GATA-4 and VEGFA.Results:There were multiple GATA transcription factor binding sites upstream of human and mouse VEGF promoters. Compared with the control group, mRNA expression of NLRP3, Caspase-1, IL-1β, GATA-4 and VEGFA in LPS group were increased (all P<0.05). Compared with LPS group, mRNA expression of NLRP3, Caspase-1, IL-1β, GATA-4 and VEGFA in LPS+ MCC950 group were significantly decreased (all P<0.05). Conclusions:NLRP3/GATA-4/VEGF signal pathway may play a significant role in the pathologic processes of nAMD.

Morphological characterization of subretinal hyper-reflective material in posterior uveitis using swept-source optical coherence tomography and optical coherence tomography angiography

Purpose: To analyze the structural features of subretinal hyper-reflective material (SHRM) in posterior uveitis using swept-source optical coherence tomography (SS-OCT) and optical coherence tomography angiography (SS-OCTA). Methods: In this observational study, subjects with quiescent posterior uveitis and the presence of SHRM on SS-OCT were subjected to SS-OCTA to identify the presence of an intrinsic choroidal neovascular (CNV) network. OCT features were compared for SHRM harboring CNV (vascular SHRM) with those without CNV network (avascular SHRM) to identify clinical signs pointing toward the presence of CNVM inside SHRM. Results: Forty-two eyes of 33 subjects (18 males; mean age: 29.52 ± 12.56 years) were evaluated. Two-thirds (28/42) of eyes having SHRM on SS-OCT harbored intrinsic neovascular network (vascular SHRM). Increased reflectivity of SHRM (P < 0.001) and increased transmission of OCT signal underlying SHRM (P = 0.03) were suggestive of the absence of CNVM. The presence of intra/subretinal fluid (P = 0.08) and pitchfork sign (P = 0.017) were important markers of vascular SHRM. Conclusion: SHRM is an important OCT finding in eyes with posterior uveitis. Meticulous assessment of SHRM characteristics on SS-OCT can aid in identifying the underlying intrinsic neovascular network.

Diagnostic value of OCTA and FFA in central serous chorioretinopathy complicated with choroidal neovascularization: a meta-analysis / 中华实验眼科杂志

Objective:To systematically evaluate the diagnostic value of optical coherence tomography angiography (OCTA) and fluorescein fundus angiography (FFA) for choroidal neovascularization (CNV) in central serous chorioretinopathy (CSC).Methods:PubMed, Cochrane Library, Web of Science, Embase, China National Knowledge Internet (CNKI), VIP database and Wanfang Database were searched for literature published from January 1991 to March 2020 with CSC patients as subjects and OCTA and FFA as diagnostic methods of CNV.The quality of the included literature was evaluated with Australian JBI tool.A meta-analysis was performed using Review Manager 5.3 software.The source of heterogeneity was identified by age and total number of samples subgroup analysis.Results:Nine studies with a total sample size of 374 eyes were enrolled.The quality score of 8 studies was greater than 14.The detection rate of CNV in CSC by OCTA was higher than that by FFA [odds ratio( OR)=3.99, 95% confidence interval( CI): 1.44-11.07, P<0.001].Studies with sample size >40 showed no heterogeneity ( I2=49%, P=0.14), suggesting that sample size might be a source of heterogeneity.Publication bias was found by funnel plot. Conclusions:OCTA has a higher detection rate of CNV secondary to CSC than FFA, and it can be used as a routine inspection method.

Effect and safety of aflibercept in the treatment of polypoidal choroidal vasculopathy with ranibizumab-resistant serous pigment epithelial detachment / 中华实验眼科杂志

Objective:To evaluate the effectiveness and safety of intravitreal injection of different doses of aflibercept for polypoidal choroidal vasculopathy (PCV) with serous pigment epithelial detachment (PED) resistant to ranibizumab.Methods:A non-randomized controlled clinical study was conducted.Seventy-three eyes of 73 patients with PCV and serous PED resistant to ranibizumab were enrolled at the First Affiliated Hospital of Zhengzhou University from January 2019 to December 2020.All patients were treated by intravitreal injection of 2 mg or 4 mg aflibercept according to patients' willingness.2 mg aflibercept or 4 mg aflibercept was intravitreally injected monthly for three consecutive months following pro re nata (PRN) regimen in 2 mg aflibercept group (38 eyes) and 4 mg aflibercept group (35 eyes), respectively.PED height and central macular thickness (CMT) were measured by optical coherence tomography, and the best corrected visual acuity (BCVA) was examined with a visual acuity chart and converted to logarithm of the minimum angle of resolution (LogMAR) unit before injection and 1 month, 2, 3, 6 months from the first injection.Intraocular pressure and treatment-related adverse events were recorded.This study adhered to the Declaration of Helsinki and was approved by an Ethics Committee of The First Affiliated Hospital of Zhengzhou University (No.2021-KY-1252).Written informed consent was obtained from each patient prior to entering study cohort.Results:Thirty-three patients (86.84%) in 2 mg aflibercept group and 30 patients (85.71%) in 4 mg aflibercept group finished the treatment and follow-up, respectively. The PED, BCVA and CMT before treatment and at the end of follow-up were (379.24±95.50) and (280.09±120.50)μm, 0.68±0.27 and 0.51±0.19, (393.96±100.81) and (291.70±44.09)μm in 2 mg aflibercept group, respectively, showing statistically significant differences (all at P<0.05).The PED, BCVA and CMT before treatment and at the end of follow-up were (393.07±93.76) and (278.63±145.07)μm, 0.66±0.31 and 0.48±0.22, (377.43±79.61) and (284.67±84.88)μm in 4 mg aflibercept group, respectively, with statistically significant differences (all at P<0.05).The CMT value in 4 mg aflibercept group was significantly lower than that in the 2 mg aflibercept group in one month after injection ( P<0.05).No severe ocular and systemic adverse events were found during the follow-up, such as retinal detachment, endophthalmitis, cataract, and persistent high intraocular pressure. Conclusions:Both 2 mg and 4 mg aflibercept can effectively treat ranibizumab-resistant PCV with serous PED, and improve the anatomical structure of retina and BCVA.4 mg aflibercept can accelerate the recovery of PED and CMT.

Critical guiding role of OCT angiography in long-term treatment and follow-up of neovascular AMD / 中华实验眼科杂志

The pathogenesis of neovascular age-related macular degeneration (nAMD) is complex and macular neovascularization (MNV), a key pathogenic factor in nAMD, is prone to recurrence.Vitreous injection of anti-VEGF drugs is the main therapy of nAMD.In recent years, a lot of progress has been made in fundus imaging techniques and optical coherence tomography angiography (OCTA) with non-invasive, rapid, stratified and high-definition functions has shown strong advantages in diagnosis, differential diagnosis, disease dynamic monitoring and follow-up of nAMD.Clinicians have had a certain understanding of the important role of OCTA in the diagnosis of nAMD and other diseases, and its clinical application value has been recognized gradually.However, its application value in follow-up of patients with nAMD and polypoid choroidal vasculopathy (PCV) is still not well understood.By reviewing a large number of recent relevant literature on OCTA, and combining the clinical practice of our research team in monitoring the course of AMD and PCV disease by OCTA, we have gained new knowledge and understanding of the pathological mechanism of AMD and PCV.In this paper, we elucidated the latest understanding of the diagnostic value of OCTA in AMD based on long-term series of OCTA studies, the new findings of OCTA in AMD management of our team, as well as its impact on ophthalmology clinical practice.Then we forecasted the role of OCTA in the prediction of recurrence and anti-VEGF treatment response, as well as the clinical value of OCTA in the optimization of nAMD treatment and follow-up plan.It is recommended that clinicians pay more attention to the clinical value and guiding role of OCTA in long-term treatment monitoring and follow-up of AMD.

Role of complement dysregulation in age-related macular degeneration / 中华实验眼科杂志

Age-related macular degeneration (AMD) is a common cause of blindness, which is still short of effective therapies.As a complex disease affected by genetical, metabolic and nutritional factors, a key factor to promote the occurrence and development of AMD is chronic inflammation.In recent years, the etiological role of abnormal complement activation in AMD has attracted lots of attention.Genetic analysis has identified a number of complement-related genes, especially CFH, affecting the susceptibility of AMD.Moreover, in vitro and in vivo studies have found that abnormal ocular and systemic complement activations are closely related to the pathological alterations of AMD, including Bruch membrane changes, drusen formation, chronic retinal inflammation and choroidal neovascularization.The dysregulation of complement activation cascades causes the damage of retinal cells, which eventually leads to the pathological changes of AMD.Accordingly, complement system has become a target for new anti-AMD therapy development.This review summarized the pathological characteristics of AMD, complement-related risk genes for AMD, and the role of abnormal complement activation in promoting the progression of AMD, so as to find new targets for AMD treatment.

Long-term follow-up of a case of choroidal neovascularization secondary to reticular pigmentary retinal dystrophy / Seguimento de longo-prazo de um caso de neovascularização de coroide secundária à distrofia reticular pigmentar da retina

ABSTRACT Reticular pigmentary retinal dystrophy, also known as Sjögren's reticular dystrophy, is a rare condition characterized by macular lesions with a reticular pattern, which are best seen on fluorescein angiogram. Choroidal neovascularization secondary to this type of dystrophy is even less common. This report describes a case of reticular pigmentary retinal dystrophy with vision loss due to neovascular membrane, which responded well to treatment with anti-vascular endothelial growth factor.

RESUMO A distrofia reticular pigmentar da retina, também conhecida como distrofia reticular de Sjögren, é uma doença rara, caracterizada por lesões maculares com um padrão reticular, que são mais bem visualizadas na angiografia com fluoresceína. A neovascularização de coroide secundária a este tipo de distrofia é ainda menos comum. Este relato descreve um caso de distrofia reticular pigmentar da retina, com perda de visão devido à membrana neovascular, que respondeu bem ao tratamento com fator de crescimento endotelial antivascular.

Progress of the application of Conbercept in ocular neovascular diseases / 国际眼科杂志(Guoji Yanke Zazhi)

Conbercept is a novel anti-vascular endothelial growth factor drug independently developed by China. Since it was approved for clinical application by the State Food and Drug Administration of China in 2013, conbercept has shown reliable safety and efficacy in the treatment of ocular neovascular diseases such as wet age-related macular degeneration, choroidal neovascularization and macular edema. For different diseases, the treatment strategies of conbercept are different. This article mainly reviews the application progress of conbercept in ocular neovascularization related diseases including wet age-related macular degeneration, diabetic macular edema, pathologic myopia choroidal neovascularization, neovascular glaucoma, retinopathy of prematurity and corneal neovascularization, and summarizes and explores the indications, administration scheme and therapeutic effect of conbercept. It is expected that the indications of conbercept will be wider and the administration scheme will be more given, and the usage of conbercept will bring new ideas for the treatment of ocular neovascular diseases.

Multimodal imaging to distinguish choroidal neovascularization from macular inflammation lesions in multifocal choroiditis / 中华眼底病杂志

Objective:To observe the multimodal image features of inflammatory lesions and choroidal neovascularization (CNV) in multifocal choroiditis (MFC).Methods:A retrospective clinical analysis. A total of 90 eyes of 46 patients with MFC diagnosed in the Department of Ophthalmology of Yunnan University Affiliated Hospital from May 2017 to April 2021 were included in the study. Among them, there were 21 males and 25 females; the average age was 38.30±8.97 years old. Twenty-nine cases of MFC were diagnosed in the past, and they visited the doctor again due to new symptoms; 17 cases without a clear past medical history were the first visits. All eyes underwent color fundus photography, fluorescein fundus angiography (FFA), optical coherence tomography (OCT), and OCT angiography (OCTA). With reference to the literature and the results of multimodal fundus imaging examinations, MFC lesions were divided into active CNV lesions, inactive CNV lesions, active inflammatory lesions, and inactive inflammatory lesions, with 31 (34.4%, 31/90), 12 (13.3%, 12/90), 26 (28.9%, 26/90), 90 (100.0%, 90/90) eyes. Nineteen eyes were treated with anti-vascular endothelial growth factor drugs. To summarize and analyze the manifestations of inflammatory lesions and CNV lesions in different imaging examinations. The Wilcoxon rank test was used to compare the detection rate of CNV lesions between FFA and OCTA.Results:In eyes with active inflammatory lesions and active CNV lesions, yellow-white lesions, retinal hemorrhage and exudation were seen on fundus color photography; FFA examination showed fluorescein leakage in the lesions; OCT examination showed retinal pigment epithelium (RPE) layer in the lesions was uplifted, the boundary was unclear, combined with subretinal and intraretinal fluid; OCTA examination showed that there was no blood flow signal in each layer of vascular tissue in active inflammatory lesions, and blood flow signals were seen in active CNV lesions. In the eyes of inactive inflammatory lesions and inactive CNV lesions, the fundus color photography showed that the lesions had clear boundaries without bleeding or exudation; FFA examination, the lesions were fluorescently stained, and there was no fluorescein leakage; OCT examination, inactive CNV lesions manifested as raised lesions with clear boundaries, and inactive inflammation manifested as scars formed by mild RPE hyperplasia or depressions in outer structures formed by atrophy; OCTA examination, inactive inflammatory lesions showed patchy loss of blood flow signal or penetrating blood flow signal below, blood flow signal can be seen in inactive CNV lesions.Conclusion:MFC active inflammatory lesions and active CNV lesions are often accompanied by retinal hemorrhage and exudation; FFA shows fluorescein leakage; OCT shows that the boundary of raised lesions is unclear; OCTA can identify the nature of CNV or inflammatory lesions.

Research progress of β-adrenergic receptor antagonists in the treatment of ocular neovascular diseases / 中华眼底病杂志

Ocular neovascularization is a pathological change in various ocular diseases such as diabetic retinopathy, retinopathy of prematurity, central retinal vein occlusion and age-related macular degeneration, which seriously affects patient's vision. β receptors are expressed in conjunctiva, corneal epithelial cells, corneal endothelial cells, extraocular muscles, trabecular meshwork, ciliary muscle, lens and retina. β adrenergic receptor antagonists bind to β receptors to exert anti-angiogenic effects by inhibiting the expression of vascular endothelial growth factor (VEGF), hypoxia-inducible factor-1, interleukin-6 and other angiogenic cytokines; reducing macrophage-related inflammatory response; increasing the expression of anti-angiogenic factors. In the treatment of corneal neovascularization, choroidal neovascularization, and retinopathy of prematurity, it can significantly reduce the area of neovascularization and delay disease progression. Co-administration of anti-VEGF drugs can reduce the frequency of administration of anti-VEGF drugs. At effective therapeutic concentrations, β-adrenergic receptor antagonists are well tolerated; they have broader targets than anti-VEGF drugs, which offers new treatment strategies for ocular neovascularization such as corneal, choroidal and retinal neovascularization.

The application and value evaluation of assisted diagnosis system for five fundus lesion based on artificial intelligence combined with optical coherence tomography / 中华眼底病杂志

Objective:To establish an artificial intelligence robot-assisted diagnosis system for fundus diseases based on deep learning optical coherence tomography (OCT) and evaluate its application value.Methods:Diagnostic test studies. From 2016 to 2019, 25 000 OCT images of 25 000 patients treated at the Eye Center of the Second Affiliated Hospital of Zhejiang University School of Medicine were used as training sets and validation sets for the fundus intelligent assisted diagnosis system. Among them, macular epiretinal membrane (MERM), macular edema, macular hole, choroidal neovascularization (CNV), and age-related macular degeneration (AMD) were 5 000 sheets each. The training set and the verification set are 18 124 and 6 876 sheets, respectively. Through the transfer learning Attention ResNet structure algorithm, the OCT image was characterized by lesion identification, the disease feature was extracted by a specific procedure, and the given image was distinguished from other types of disease according to the statistical characteristics of the target lesion. The model algorithms of MERM, macular edema, macular hole, CNV and AMD were initially formed, and the fundus intelligent auxiliary diagnosis system of five models was established. The performance of each model-assisted diagnosis in the fundus intelligent auxiliary diagnostic system was evaluated by applying the subject working characteristic curve, area under the curve (AUC), sensitivity, and specificity.Results:With the intelligent auxiliary diagnosis system, the diagnostic sensitivity of the MERM was 93.5%, the specificity was 99.23%, and AUC was 0.983 7; the diagnostic sensitivity of macular edema was 99.02%, the specificity was 98.17%, and AUC was 0.994 6; the diagnostic sensitivity of macular hole was 98.91%, the specificity was 99.91%, AUC was 0.996 2; the diagnostic sensitivity of CNV was 97.54%, the specificity was 94.71%, AUC was 0.987 5; the diagnostic sensitivity of AMD was 95.12%, the specificity was 97.09%, AUC was 0.985 3.Conclusions:The artificial intelligence robot-assisted diagnosis system for fundus diseases based on deep learning for OCT images has accurate and efficient diagnostic performance for assisting the diagnosis of MERM, macular edema, macular hole, CNV, and AMD.

Intravitreal Conbercept or Ranibizumab for myopic choroidal neovascularization / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To compare the efficacy of intravitreal conbercept or ranibizumab for myopic choroidal neovascularization(CNV).<p>METHODS: A retrospective cohort study. This study included 46 patients(46 eyes)with myopic CNV who were treated with conbercept(conbercept group, 20 cases, 20 eyes)or ranibizumab(ranibizumab group, 26 cases, 26 eyes)from March 2015 to August 2019. Central macular thickness(CMT), the number of injections and complications measured by best-corrected visual acuity(BCVA)and optical coherence tomography(OCT)were compared between the two groups before treatment and 1, 3, 6mo after treatment.<p>RESULTS: Before treatment, the BCVA(LogMAR)of conbercept and ranibizumab groups were 0.81±0.51, 0.83±0.66(<i>P</i>=0.900). After treatment, the BCVA(LogMAR)in the conbercept group at 1, 3 and 6mo were 0.59±0.33, 0.49±0.34, 0.44±0.32, in the ranibizumab group were 0.53±0.54, 0.47±0.47, 0.40±0.43. The BCVA was significantly improved in both groups after treatment(all <i>P</i><0.001). Before treatment, the CMT of conbercept and ranibizumab groups were 242.30±73.27, 233.38±66.63μm(<i>P</i>=0.669). After treatment, the CMT in the conbercept group at 1, 3, and 6mo were 217.00±54.78, 208.65±55.38, 206.00±45.34μm, in the ranibizumab group were 197.42±50.47, 198.38±55.19, 192.15±51.97μm. The CMT was significantly decreased in both groups after treatment(all <i>P</i><0.05). There were no significant differences in the number of injections, BCVA and CMT at each follow-up time points between conbercept and ranibizumab groups(all <i>P</i>>0.05). Systemic adverse reactions and serious ocular complications were not found during the treatment period.<p>CONCLUSION: Intravitreal conbercept or ranibizumab provide similar efficacy to improve the BCVA and reduce the CMT in the patients with myopic CNV. Both conbercept and ranibizumab could be a choice of treatment for myopic CNV.

Study on influence factors of intravitreal Ranibizumab in chronic central serous chorioretinopathy combined with CNV / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To observe the efficacy of intravitreal anti-vascular endothelial growth factor(VEGF)injection treatment in chronic central serous chorioretinopathy(CCSC)combined with choroidal neovascularization(CNV)using multimodal imaging, to explore and evaluate the influence factors.<p>METHODS: In this retrospective study, 30 patients(30 eyes)were diagnosed as CCSC combined with CNV. Comprehensive ophthalmologic examinations were performed, including best corrected visual acuity(BCVA), enhanced-depth imaging(EDI)spectral domain optical coherence tomography(SD-OCT), fundus fluorescein angiography(FFA), Indocyanine green angiography(ICGA), and optical coherence tomography angiography(OCTA). Patients were treated with intravitreal ranibizumab(IVR)parallel 1+PRN schem for subretinal fluid(SRF)secondary to CCSC combined with CNV. All the patients were followed up at 1wk, 1mo after treatment and 3mo after consecutive treatment. The BCVA, central macular thickness(CMT), subfoveal choroid thickness(SFCT)and CNV flow area were compared.<p>RESULTS: All the patients were observed at baseline, 1wk, 1mo after treatment and 3mo after consecutive treatment. The difference at various time points of CMT(μm)were statistically significant(<i>F</i>=62.06, <i>P</i><0.01). CMT after treatment at each time point was compared with baseline, the difference among each time points was statistically significant(<i>t</i>=3.08, 6.57, 4.90; <i>P</i>=0.01, 0.02, <0.01). In which 46.7% of patients, SRF can be completely absorbed(14/30). The difference at various time points of BCVA(LogMAR)were statistically significant(<i>F</i>=87.21, <i>P</i><0.01). BCVA after treatment at each time point was compared with baseline, the difference between each group was statistically significant(<i>t</i>=6.52, 4.71, 6.01; <i>P</i>=0.03, <0.01, <0.01). The difference at various time points of SFCT(μm)were statistically significant(<i>F=</i>57.98, <i>P</i><0.01). SFCT after treatment at each time point was compared with baseline, the difference among each time points was statistically significant(<i>t</i>=5.11, 9.03, 4.2; <i>P</i>=0.03, <0.01, <0.01). The difference at various time points of CNV area(mm²)were statistically significant(<i>F</i>=70.78, <i>P</i><0.01). CNV area at 1wk and 1mo after treatment was compared with baseline, the difference was no statistically significant(<i>t</i>=7.01, 6.54; <i>P</i>=0.07, 0.05). CNV area at 3mo after the last treatment was compared with baseline, the difference was statistically significant(<i>t</i>=4.51, <i>P</i>=0.02). The change of CMT was positively correlated with the baseline CMT, BCVA and CNV area(<i>r</i>=0.43, 0.41, 0.41; <i>P=</i>0.02, 0.03, 0.03). The change of BCVA was positively correlated with the baseline BCVA and CMT(<i>r</i>=0.89, 0.43; <i>P</i><0.01, 0.02). <p>CONCLUSION: CCSC combined with CNV show different sensitivity to anti-VEGF therapy, the SRF can be completely absorbed after treatment in parts of patients. CNV may not be the only predictive factor leading to the SRF. The baseline BCVA, CMT and CNV area may be the factors that influence the final therapeutic effect of the intravitreal anti-VEGF injection therapy.

Imaging features of OCTA in the macular hemorrhage of pathologic myopia / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To observe the imaging features of optical coherence tomography angiography(OCTA)in the macular hemorrhage of pathologic myopia.METHODS:Designing a retrospective analysis collected clinical data of 100 patients(108 eyes)diagnosed as macular hemorrhage of pathological myopic in Nanjing Medical University Affiliated Eye Hospital from June 2016 to December 2020. All patients underwent refraction, eye axis,fundus photography, spectral-domain optical coherence tomography(SD-OCT), fundus fluorescein angiography(FFA), indocyanine green angiography(ICGA)and OCTA examination. All patients were divided into macular hemorrhage only with lacquer cracks and macular hemorrhage with choroidal neovascularization(CNV). All patients followed-up for more than 3mo by OCTA. RESULTS:There were 40 patients(42 eyes)diagnosed as macular hemorrhage only with lacquer cracks, OCTA showed bleed obscured by choroidal capillaries. After hemorrhage was being absorbed, lacquer cracks showed linear or stellate reflection completely in the choroidal capillary layer. B-scan image showed discontinuous retinal pigment epithelium(RPE), thinner choroid and an increased light. Penetrance into deeper tissues. After all macular hemorrhage only with lacquer cracks were absorbed, follow-up mode of OCTA found that 2 eyes(4.8%)without lacquer cracks, 28 eyes(66.7%)were linear and 12 eyes(28.6%)were stellate under the original hemorrhage. Follow-up mode also showed that 8 eyes of 8 patients(19.0%)relapsed macular hemorrhage only with lacquer cracks, and 4 eyes of 4 patients(9.5%)suffered secondary macular hemorrhage with CNV. There were 60 patients(66 eyes)diagnosed as macular hemorrhage with CNV,OCTA showed bleed obscured choroidal capillaries, the outer retinal and choroidal capillary layer also showed the shape of CNV around hemorrhage. B-scan showed CNV breaked through the RPE layer and blood flow signal in it. The area of CNV decreased after anti-vascular endothelial growth factor(VEGF)intravitreal injection treatment. Around all macular hemorrhage with CNV, OCTA found that 48 eyes(72.7%)had lacquer cracks, 28 eyes(42.4%)were linear and 20 eyes(30.3%)were stellate.CONCLUSION:OCTA has a great significance in the diagnosis of macular hemorrhage of pathological myopia, fast and non-invasive is the biggest advantage. Choroidal capillary layer can clearly observe the shape and location of hemorrhage,lacquer cracks and CNV. The follow-up mode can intuitively comprehend the changes of disease. To some extent, it can replace fundus angiography to directly judge the classification, and time to treat in the clinic.

Mechanism study of Curcumin on inhibiting experimental choroidal neovascularization <i>in vitro</i> by AKT/HIF-1α/VEGF pathway / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the effect and mechanism of curcumin on inhibiting choroidal neovascularization(CNV)<i>in vitro</i>. METHODS: Human retinal pigment epithelial(ARPE-19)cells chemical hypoxia model was established by cobalt chloride(CoCl2). CCK-8 method was used to detect the effect of curcumin on the activity of ARPE-19 cells induced by CoCl2. RT-qPCR and Western blot were used to detect the expression of AKT, HIF-1α, VEGF mRNA and protein in ARPE-19 cells hypoxia model induced by CoCl2. Cell scratch test, transwell chamber migration test, transwell chamber invasion test and matrigel matrix hose lumen formation test were used to observe the effects of conditioned medium of curcumin in ARPE-19 cells on the proliferation, migration, invasion and lumen formation of human umbilical vein endothelial cells(HUVEC)in non-contact condition. RESULTS:Chemical hypoxia model of ARPE-19 cells can successfully establish by CoCl2 at 100μmol/L. CoCl2 at the final concentration of 100μmol/L can promote the expression of AKT, HIF-1α and VEGF mRNA and p-AKT, HIF-1α and VEGF protein in ARPE-19 cells. Curcumin at the final concentration of 100μmol/L can reduce the expression of AKT, HIF-1α and VEGF mRNA in ARPE-19 hypoxia model. Curcumin at the final concentration of 100μmol/L can reduce the expression of AKT, HIF -1α and VEGF proteins in ARPE-19 hypoxia model. The conditioned medium of low(6.25μmol/L), medium(25μmol/L)and high dose(100μmol/L)curcumin in ARPE-19 cells can significantly inhibit the level migration of HUVEC. The conditioned medium in high dose group can significantly inhibit the vertical migration and cell invasion of HUVEC. The conditioned medium of middle and high dose curcumin in ARPE-19 cells can inhibit the lumen formation of HUVEC. CONCLUSION:Curcumin at 100μmol/L can protect ARPE-19 cells from hypoxia induced by CoCl2. Curcumin can inhibit the formation of blood vessels at the cellular level.