Monogenic gene therapy for glaucoma and optic nerve injury

The prevalence of glaucoma, the second leading cause of global blindness, is increasing due to aging populations. In glaucoma, degeneration of the optic nerve and retinal ganglion cells(RGCs) causes visual field defects and eventual blindness.

Practice patterns among ophthalmic surgeons in treating concomitant oculoplastic conditions in preoperative period:A questionnaire-based study

BACKGROUND Addressing oculoplastic conditions in the preoperative period ensures both the safety and functional success of any ophthalmic procedure.Some oculoplastic conditions,like nasolacrimal duct obstruction,have been extensively studied,whereas others,like eyelid malposition and thyroid eye disease,have received minimal or no research.AIM To investigate the current practice patterns among ophthalmologists while treating concomitant oculoplastic conditions before any subspecialty ophthalmic intervention.METHODS A cross-sectional survey was disseminated among ophthalmologists all over India.The survey included questions related to pre-operative evaluation,anaesthetic and surgical techniques preferred,post-operative care,the use of adjunctive therapies,and patient follow-up patterns.RESULTS A total of 180 ophthalmologists responded to the survey.Most practitioners(89%)felt that the ROPLAS test was sufficient during pre-operative evaluation before any subspecialty surgery was advised.The most common surgical techniques employed were lacrimal drainage procedures(Dacryocystorhinostomy)(63.3%),eyelid malposition repair(36.9%),and ptosis repair(58.7%).Post-operatively,47.7%of respondents emphasized that at least a 4-week gap should be maintained after lacrimal drainage procedures and eyelid surgeries.Sixty-seven percent of ophthalmologists felt that topical anaesthetic procedures should be preferred while performing ocular surgeries in thyroid eye disease patients.CONCLUSION Approximately 50%of ophthalmologists handle prevalent oculoplastic issues themselves,seeking the expertise of an oculoplastic surgeon under particular conditions.Many ophthalmologists still favor using ROPLAS as a preliminary screening method before proceeding with cataract surgery.Eyelid conditions and thyroid eye disease are not as commonly addressed before subspecialty procedures compared to issues like nasolacrimal duct obstruction and periocular infections.

REG增强的脱细胞猪角膜/聚甲基丙烯酸羟乙酯原位一体式全层复合人工角膜

背景:目前用于全层移植的人工角膜缺乏生物活性及力学适配性,组合式人工角膜存在镜柱和周围组分间的界面问题。目的:在脱细胞猪角膜原位固化制备具有多肽增强、匹配自然角膜机械强度、良好透光性的一体式全层人工角膜。方法:使用非离子型脱细胞试剂Triton X-100与超声冻融及超级核酸酶相结合的方法制备脱细胞猪角膜,将聚甲基丙烯酸羟乙酯单体与光引发剂同时引入脱细胞猪角膜中,通过紫外滤光片遮住除中心区域以外的部分,使用275 nm紫外光引发中央区域聚合,除去未反应的单体及引发剂后得到中央光学区,同理在后板层固化隔水区,最后引入REG活性多肽,得到原位一体式全层人工角膜,表征人工角膜的物理性能、力学性能、透光性、降解性能及体内外生物相容性。结果与结论:①实验在脱细胞猪角膜的中央区域采用聚甲基丙烯酸羟乙酯原位构建了一个具有聚合物和胶原纤维共存的光学区,扫描电镜下可见人工角膜的上表面较为粗糙且轮廓不规则,具有明显的凹陷和突起结构,下表面相对光滑;该人工角膜具有接近于天然角膜的力学性能,光学区透光率达到自然角膜的80%,浸泡于含胶原酶的PBS无菌溶液中可较好地保留固化光学区和隔水区,维持角膜的基本结构;该人工角膜具有良好的细胞相容性,能够为细胞提供适宜的黏附生长环境,有利于角膜上皮细胞的迁移和黏附,促进血管内皮细胞的生长和新生血管的形成,促进上皮化过程;人工角膜植入SD大鼠皮下12周后具有良好的生物相容性和安全性,可降低植入初期的急性炎症反应;②结果表明,实验制备的一体式全层人工角膜具有作为全层人工角膜支架材料的潜力。

丹芪化瘀方含药血清对光损伤人视网膜色素上皮细胞保护机制研究

目的:探讨丹芪化瘀方含药血清对LED冷光源致人视网膜色素上皮细胞(ARPE-19)光损伤的防治作用及最佳药物剂量,探寻其可能存在的保护机制。方法:应用健康的SD大鼠制备丹芪化瘀方空白血清和含药血清,将0%(空白血清)及5%、10%、15%、20%含药血清作用于ARPE-19细胞,每组设6个复孔。各组加入相应的药物于细胞恒温培养箱中培养24 h后采用CCK-8检测细胞活力,筛选最佳含药血清浓度进行后续实验。实验设置对照组、空白血清组、低剂量、中剂量、高剂量含药血清组培养,采用实时荧光定量聚合酶链式反应(Real-time PCR)检测细胞中诱导型一氧化氮合酶(iNOS)、内皮型一氧化氮合酶(eNOS)、内皮素-1(ET-1)mRNA表达;酶联免疫吸附测定法(ELISA)检测一氧化氮合成酶(NOS)、一氧化氮(NO)、血管内皮生长因子(VEGF)含量;蛋白免疫印迹法(Western blot)检测对照组、空白血清组、高剂量组iNOS的蛋白表达水平。结果:筛选出16%丹芪化瘀方含药血清作为后续实验干预浓度。与对照组比较,空白血清组iNOS、ET-1 mRNA表达及NOS、NO、VEGF含量均升高(P<0.001),eNOS mRNA表达降低(P<0.001);随着含药血清浓度的增加细胞中iNOS、ET-1 mRNA表达及NOS、NO、VEGF含量逐渐降低,eNOS mRNA表达增加(P<0.001),与低、中剂量组相比,高剂量组的iNOS、ET-1 mRNA表达及NOS、NO、VEGF含量显著降低(P<0.001或P<0.01或P<0.05),eNOS mRNA显著升高(P<0.001或P<0.05);空白血清组iNOS蛋白水平明显高于对照组,而16%含药血清的iNOS蛋白水平明显低于空白血清组(P<0.001)。结论:丹芪化瘀方含药血清可能通过抑制iNOS/NO途径减轻细胞损伤,上调eNOS表达,下调ET-1、VEGF的表达,抑制脉络膜新生血管形成并改善血-视网膜屏障功能,从而发挥保护作用。

重视眼病前期的流行病学调查

流行病学调查是疾病早预防、早干预以及相关政策制定重要的科学依据。疾病前期(predisease)常指在流行病学调查/筛查中发现的机体非正常状态,但还没有达到疾病的诊断标准,是机体由健康状态向疾病状态进展的过渡阶段,如果不干预就会发展为疾病。如大家熟知的癌症前期(癌前病变)、糖尿病前期以及高血压前期等,也包括近视前期、青光眼前期以及糖尿病前期眼部并发症等眼病前期。疾病前期由于处于疾病尚未出现临床表现或者极早期阶段,通过适当的干预可以阻止甚至逆转疾病的发生而广受重视。然而,目前眼科工作者尚未对开展眼病前期流行病学调查足够重视,对如何开展眼病前期的流行病学调查也不甚了解,本文根据以往开展眼病流行病学调查的经验,就目前患病率高、视觉损伤大的几种常见眼病前期流行病学调查的开展进行论述。

2013—2022年北京市海淀区学龄前儿童眼保健及视力低常状况分析

目的:了解北京市海淀区幼儿园4~6岁在园儿童眼保健状况及其变化趋势,为制定相应干预措施提供依据。方法:对2013—2022年北京市海淀区幼儿园4~6岁儿童眼保健年报数据进行分析。结果:2013—2022年海淀区幼儿园4~6岁儿童视力检查率为99.16%~99.89%,呈逐年上升趋势,差异有统计学意义(χ^(2)=238.628,P<0.01);视力低常患病率为5.97%~7.57%,整体呈上升趋势,差异有统计学意义(χ^(2)=17.355,P<0.01),2021年达到最高值,2022年降为最低;弱视发生率为0.41%~1.29%,呈逐年下降趋势,差异有统计学意义(χ^(2)=451.419,P<0.01);视力低常矫治率为75.74%~97.63%,呈逐年上升趋势,差异有统计学意义(χ^(2)=1 087.365,P<0.01)。本地儿童视力低常率和视力低常矫治率均高于外地户籍儿童。结论:北京市海淀区学龄前儿童眼保健覆盖状况良好,儿童视力低常率呈逐年上升趋势,本市户籍儿童是预防发生视力低常的重点人群,外地户籍儿童家长应加强对儿童视力低常矫治的重视,可以通过加强针对家长和教师的眼保健健康教育,注重培养儿童良好用眼习惯等综合干预措施降低儿童视力低常检出率。

0~6岁儿童眼保健和视力检查服务现状及对策研究

目的了解0~6岁儿童眼保健和视力检查服务现状,为相关部门提供决策依据。方法采用方便抽样方法,于2019年9—12月从北京市、安徽省、青海省抽取15家区县级妇幼保健院及基层卫生机构进行问卷调查,并采用现场观察及小组访谈的方式了解儿童眼保健服务现状及存在的问题等。结果被调查的区县级妇幼保健机构均配备了开展儿童眼保健服务的基本用房及设备;基层卫生机构均配备了开展儿童眼保健服务的基础设备,但有些设备陈旧破损、闲置未用。被调查机构儿童眼保健人员均不足,区县级妇幼保健院、社区卫生服务中心和乡镇卫生院的儿童眼保健人员分别占卫生技术人员的9.5%、5.5%和2.4%,不同类型卫生机构的儿童眼保健人员占比差异有统计学意义(χ^(2)=8.966,P=0.011)。社区卫生服务中心无眼科医生,该岗位由儿保人员兼职。被调查机构儿童眼保健人员接受上级指导及培训的机会普遍较少,年人均1~3次。各地儿童眼保健及视力检查服务覆盖率均在90%以上,但现场观察发现儿童眼保健人员操作技能尚不熟练、不规范。结论各卫生机构均需进一步完善儿童眼保健及视力检查服务设备,提高儿童眼保健人员的知识技能及服务质量。

慢性泪囊炎合并睑板腺功能障碍综合治疗后疗效评价

目的:探讨慢性泪囊炎合并睑板腺功能障碍患者通过泪道激光成型并置管术及术后联合睑板腺治疗的疗效。方法:收集我院2021-03/2022-12慢性泪囊炎合并睑板腺功能障碍的患者128例,均行泪道激光成型联合置管术;术后随机分为2组,A组64例(无睑板腺治疗)、B组64例(睑板腺治疗),术后3 mo取出泪道置管后评价两组泪道冲洗通畅率;并对比两组眼表疾病指数(OSDI)评分、非侵袭性泪膜破裂时间、泪河高度、眼红分析、睑板腺分析、泪液脂质层厚度、泪液蕨类试验、结膜印记细胞检查的变化情况。结果:A组与B组泪道冲洗通畅率分别为78.1%、81.2%,两者比较无差异(P>0.05);与A组比较,术后3 mo B组患者非侵袭性泪膜破裂时间延长,OSDI评分、眼红分析、泪液蕨类试验及结膜印记细胞学分级降低(均P<0.05),而泪河高度、泪液脂质层厚度、睑板腺缺失评分两者无明显差异(均P>0.05)。结论:慢性泪囊炎合并睑板腺功能障碍患者综合治疗后其舒适度、泪膜稳定性、局部炎症反应有所好转,这类患者手术治疗的同时应同步重视眼表微环境异常的改善,以期达到满意的疗效。

西安市中小学生视力筛查和戴镜情况分析

目的:了解西安市中小学生视力和戴镜情况,为开展近视防控工作提供科学依据。方法:在西安市16个区县对119所中小学校38 226名学生开展视力筛查和戴镜情况调查,对裸眼视力、屈光度、戴镜率、足矫率等进行统计分析。结果:西安市中小学近视率为61.53%,随着学段增加而递增(χ^(2)_(趋势)=5332.203,P<0.01),其中轻度近视率占比随着学段的升高而降低,中度、高度占比随着学段的升高呈上升趋势(χ^(2)_(趋势)=2671.562,P<0.01)。近视学生戴镜率为51.69%,戴镜率随着学段的升高呈上升趋势(χ^(2)_(趋势)=1486.941,P<0.01),女生戴镜率高于男生(χ^(2)=23.659,P<0.01),城区戴镜率高于郊县(χ^(2)=102.241,P<0.01)。戴镜学生足矫率为67.08%,城区戴镜学生足矫率高于郊县(χ^(2)=4.980,P<0.05)。近视学生过去1 a内视力检查频次≥2次的占63.66%,职业高中学生视力检查频次≥2次占58.06%。近视学生居住在郊县与戴镜率呈负相关,学段升高、每年视力检查频次增多与近视学生戴镜率呈正相关(均P<0.01)。结论:西安市学生近视防控形势严峻,学生戴镜率、足矫率、视力检查频次低,需要重点关注郊县、小学、职业高中学生视力矫正情况。

环孢素A在眼表疾病中的应用及进展

环孢素A是一种具有强免疫抑制活性的环状多肽,作用机制广泛,包括免疫抑制作用、抗炎、抑制细胞凋亡、促进上皮愈合及杯状细胞功能恢复和增加泪液分泌,与修复眼表疾病的损伤密切相关。由于其疗效显著、抑制疾病复发且副作用较小,近年来,越来越多的眼表疾病如干眼、角膜移植术后排斥反应、春季角结膜炎、非感染性角膜炎、单纯疱疹病毒性角膜基质炎等在临床中使用环孢素A。然而,不同浓度和剂型的环孢素A对眼表炎症性疾病的治疗具有选择性差异。因此,为了系统地了解环孢素A对眼表疾病的影响,文章对其展开综述。

Age-related driving mechanisms of retinal diseases and neuroprotection by transcription factor EB-targeted therapy

Retinal aging has been recognized as a significant risk factor for various retinal disorders,including diabetic retinopathy,age-related macular degeneration,and glaucoma,following a growing understanding of the molecular underpinnings of their development.This comprehensive review explores the mechanisms of retinal aging and investigates potential neuroprotective approaches,focusing on the activation of transcription factor EB.Recent meta-analyses have demonstrated promising outcomes of transcription factor EB-targeted strategies,such as exercise,calorie restriction,rapamycin,and metformin,in patients and animal models of these common retinal diseases.The review critically assesses the role of transcription factor EB in retinal biology during aging,its neuroprotective effects,and its therapeutic potential for retinal disorders.The impact of transcription factor EB on retinal aging is cell-specific,influencing metabolic reprogramming and energy homeostasis in retinal neurons through the regulation of mitochondrial quality control and nutrient-sensing pathways.In vascular endothelial cells,transcription factor EB controls important processes,including endothelial cell proliferation,endothelial tube formation,and nitric oxide levels,thereby influencing the inner blood-retinal barrier,angiogenesis,and retinal microvasculature.Additionally,transcription factor EB affects vascular smooth muscle cells,inhibiting vascular calcification and atherogenesis.In retinal pigment epithelial cells,transcription factor EB modulates functions such as autophagy,lysosomal dynamics,and clearance of the aging pigment lipofuscin,thereby promoting photoreceptor survival and regulating vascular endothelial growth factor A expression involved in neovascularization.These cell-specific functions of transcription factor EB significantly impact retinal aging mechanisms encompassing proteostasis,neuronal synapse plasticity,energy metabolism,microvasculature,and inflammation,ultimately offering protection against retinal aging and diseases.The review emphasizes transcription factor EB as a potential therapeutic target for retinal diseases.Therefore,it is imperative to obtain well-controlled direct experimental evidence to confirm the efficacy of transcription factor EB modulation in retinal diseases while minimizing its risk of adverse effects.

Subretinal fibrosis secondary to neovascular age-related macular degeneration:mechanisms and potential therapeutic targets

Subretinal fibrosis is the end-stage sequelae of neovascular age-related macular degeneration.It causes local damage to photoreceptors,retinal pigment epithelium,and choroidal vessels,which leads to permanent central vision loss of patients with neovascular age-related macular degeneration.The pathogenesis of subretinal fibrosis is complex,and the underlying mechanisms are largely unknown.Therefore,there are no effective treatment options.A thorough understanding of the pathogenesis of subretinal fibrosis and its related mechanisms is important to elucidate its complications and explore potential treatments.The current article reviews several aspects of subretinal fibrosis,including the current understanding on the relationship between neovascular age-related macular degeneration and subretinal fibrosis;multimodal imaging techniques for subretinal fibrosis;animal models for studying subretinal fibrosis;cellular and non-cellular constituents of subretinal fibrosis;pathophysiological mechanisms involved in subretinal fibrosis,such as aging,infiltration of macrophages,different sources of mesenchymal transition to myofibroblast,and activation of complement system and immune cells;and several key molecules and signaling pathways participating in the pathogenesis of subretinal fibrosis,such as vascular endothelial growth factor,connective tissue growth factor,fibroblast growth factor 2,platelet-derived growth factor and platelet-derived growth factor receptor-β,transforming growth factor-βsignaling pathway,Wnt signaling pathway,and the axis of heat shock protein 70-Toll-like receptors 2/4-interleukin-10.This review will improve the understanding of the pathogenesis of subretinal fibrosis,allow the discovery of molecular targets,and explore potential treatments for the management of subretinal fibrosis.

Small extracellular vesicles derived from human induced pluripotent stem cell-differentiated neural progenitor cells mitigate retinal ganglion cell degeneration in a mouse model of optic nerve injury

Several studies have found that transplantation of neural progenitor cells(NPCs)promotes the survival of injured neurons.However,a poor integration rate and high risk of tumorigenicity after cell transplantation limits their clinical application.Small extracellular vesicles(sEVs)contain bioactive molecules for neuronal protection and regeneration.Previous studies have shown that stem/progenitor cell-derived sEVs can promote neuronal survival and recovery of neurological function in neurodegenerative eye diseases and other eye diseases.In this study,we intravitreally transplanted sEVs derived from human induced pluripotent stem cells(hiPSCs)and hiPSCs-differentiated NPCs(hiPSC-NPC)in a mouse model of optic nerve crush.Our results show that these intravitreally injected sEVs were ingested by retinal cells,especially those localized in the ganglion cell layer.Treatment with hiPSC-NPC-derived sEVs mitigated optic nerve crush-induced retinal ganglion cell degeneration,and regulated the retinal microenvironment by inhibiting excessive activation of microglia.Component analysis further revealed that hiPSC-NPC derived sEVs transported neuroprotective and anti-inflammatory miRNA cargos to target cells,which had protective effects on RGCs after optic nerve injury.These findings suggest that sEVs derived from hiPSC-NPC are a promising cell-free therapeutic strategy for optic neuropathy.

The gut-eye axis:from brain neurodegenerative diseases to age-related macular degeneration

Age-related macular degeneration is a serious neurodegenerative disease of the retina that significantly impacts vision.Unfortunately,the specific pathogenesis remains unclear,and effective early treatment options are consequently lacking.The microbiome is defined as a large ecosystem of microorganisms living within and coexisting with a host.The intestinal microbiome undergoes dynamic changes owing to age,diet,genetics,and other factors.Such dysregulation of the intestinal flora can disrupt the microecological balance,resulting in immunological and metabolic dysfunction in the host,and affecting the development of many diseases.In recent decades,significant evidence has indicated that the intestinal flora also influences systems outside of the digestive tract,including the brain.Indeed,several studies have demonstrated the critical role of the gut-brain axis in the development of brain neurodegenerative diseases,including Alzheimer’s disease and Parkinson’s disease.Similarly,the role of the“gut-eye axis”has been confirmed to play a role in the pathogenesis of many ocular disorders.Moreover,age-related macular degeneration and many brain neurodegenerative diseases have been shown to share several risk factors and to exhibit comparable etiologies.As such,the intestinal flora may play an important role in age-related macular degeneration.Given the above context,the present review aims to clarify the gut-brain and gut-eye connections,assess the effect of intestinal flora and metabolites on age-related macular degeneration,and identify potential diagnostic markers and therapeutic strategies.Currently,direct research on the role of intestinal flora in age-related macular degeneration is still relatively limited,while studies focusing solely on intestinal flora are insufficient to fully elucidate its functional role in age-related macular degeneration.Organ-on-a-chip technology has shown promise in clarifying the gut-eye interactions,while integrating analysis of the intestinal flora with research on metabolites through metabolomics and other techniques is crucial for understanding their potential mechanisms.

Postnatal development of rat retina:a continuous observation and comparison between the organotypic retinal explant model and in vivo development

The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and continuous comparison between in vivo retinal development and the organotypic retinal explant culture makes this model controversial in postnatal retinal development studies.Thus,we aimed to verify the feasibility of using this model for postnatal retinal development studies by comparing it with the in vivo retina.In this study,we showed that postnatal retinal explants undergo normal development,and exhibit a consistent structure and timeline with retinas in vivo.Initially,we used SOX2 and PAX6 immunostaining to identify retinal progenitor cells.We then examined cell proliferation and migration by immunostaining with Ki-67 and doublecortin,respectively.Ki-67-and doublecortin-positive cells decreased in both in vivo and explants during postnatal retinogenesis,and exhibited a high degree of similarity in abundance and distribution between groups.Additionally,we used Ceh-10 homeodomain-containing homolog,glutamate-ammonia ligase(glutamine synthetase),neuronal nuclei,and ionized calcium-binding adapter molecule 1 immunostaining to examine the emergence of bipolar cells,Müller glia,mature neurons,and microglia,respectively.The timing and spatial patterns of the emergence of these cell types were remarkably consistent between in vivo and explant retinas.Our study showed that the organotypic retinal explant culture model had a high degree of consistency with the progression of in vivo early postnatal retina development.The findings confirm the accuracy and credibility of this model and support its use for long-term,systematic,and continuous observation.

AAV2-PDE6B restores retinal structure and function in the retinal degeneration 10 mouse model of retinitis pigmentosa by promoting phototransduction and inhibiting apoptosis

Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.