后发性白内障的发生机制及防治研究现状

后囊膜混浊(posterior capsule opacification,PCO)又称后发性白内障,是现代白内障囊外摘除术后最常见的并发症。后发性白内障的发生与患者手术时的年龄,囊袋的大小,囊口的连续性,血房水屏障破坏释放的炎症介质,残留的晶状体上皮细胞及晶状体皮质的多少,以及所植入的人工晶状体的材质等。研究者对手术方式的改进及人工晶体材料设计进行研究,以降低白内障囊外摘除术后后囊膜混浊的发生率,但它仍然是一个重大的没有解决的难题。Nd:YAG激光后囊膜切开术是目前治疗PCO最常用的有效方法,但仍会出现一定的并发症,并且治疗费用较高。研究者进一步改进手术技术,人工晶体材料设计和在使用药物治疗方面,选择即安全有效和较少的费用消除PCO。     

飞秒激光与生物细胞作用机理及应用

飞秒激光是自1960年第一台激光器诞生以来,过去20年间由激光科学发展起来的最强有力的新工具之一。飞秒激光由于脉冲持续时间短、瞬时功率大、聚焦尺寸小的特点,使得其在超快、超强和超精细领域有着广阔的应用前景。其中最重要的一个方向是飞秒激光在生物细胞方面的应用。细胞是生命活动的基本单位。所有的病源微观上都体现在细胞中细胞器的工作,所以用飞秒激光作用在病体的细胞器上达到治疗的目的,是一个很有前景的领域。由于生物大分子和水几乎不吸收近红外光,故应用近红外飞秒激光对细胞进行手术,同时可在不损伤细胞活性的前提下对细胞进行实验。这种激光手术技术已被用于对细胞内结构进行切割和蚀除。介绍了该技术在细胞领域中的一些应用,如纳米手术、基因转染和染色体切割等;还介绍了飞秒激光技术与生物细胞中主要细胞器的祛除的原理、飞秒激光细胞操作与手术系统和实验中荧光成像、多光子成像显微镜等手段。     

干细胞与晶状体再生

白内障摘除联合人工晶状体植入术是目前治疗白内障的唯一有效措施。然而,人工晶状体作为替代材料,仍然存在一些如屈光调节力差以及术后眩光等未能克服的缺陷。寻找更理想的晶状体替代物及低等两栖类动物（如蝾螈）强大的晶状体再生能力,为晶状体再生的研究提供了原动力和依据。近年来,人们已探索出将胚胎干细胞/诱导的多能干细胞在体外诱导分化为类晶状体样结构的培养方法,为白内障的治疗开辟了新的思路。晶状体再生的研究为探索晶状体正常发育机制及晶状体疾病的发生和防治提供了新的平台。晶状体再生的成功也将为白内障的防治带来里程碑性的突破。本文拟总结晶状体正常发育过程及其调控机制,回顾国内外对晶状体体内再生能力的研究成果,并对目前人们探索利用胚胎干细胞和诱导的多能干细胞再造晶状体的研究进展作一概述,希望对干细胞与晶状体再生的后续相关研究提供一定的借鉴。     

羊毛甾醇可以减轻白内障

<正>白内障(cataract)是由于人眼晶状体(lens)出现混浊,使视力发生障碍,影响成像质量的疾病,是目前全球第一大致盲性眼病。手术摘除混浊的晶状体是目前唯一可以治愈白内障,使患者重获光明的治疗手段。长期以来,开发可以治疗白内障的眼药水是眼科医生的梦想。晶状体内的(crystallin)对于保持晶状体的透明性至关重要,白内障的发生主要由晶状体蛋白的聚集引起。前人研究发     

波形蛋白与糖皮质激素性白内障发病关系的研究进展

长期全身或局部应用糖皮质激素较易导致晶状体后囊膜混浊,形成糖皮质激素性白内障,机理尚不明确。随着糖皮质激素在器官移植、免疫系统障碍、过敏及创伤救治等治疗过程中的大量应用,激素性白内障的发生率也大幅上升,因此,糖皮质激素性白内障也越来越受到人们的重视。目前对于糖皮质激素性白内障的发病机制尚不明确,主流的包括氧化-构象学说、蛋白复合物形成学说、细胞粘附分子异常学说及糖皮质激素作用于糖皮质激素受体从而发生改变的学说。激素发生改变的方法也许同晶状体蛋白共同作用或者协同作用均相关,就像通过细胞调控、粘附调节、受体等协同作用发生改变。而以往的研究表明波形蛋白在晶状体上皮细胞中正确和准确的存在,可起着保持晶状体细胞基本状态的首要作用。同时波形蛋白在晶状体里的反应会导致晶状体上皮细胞的变化,及晶状体细胞基本状态的变化。近来研究发现,糖皮质激素受体介导的晶状体波形蛋白的改变参与了激素性白内障的形成。     

PDGF-A在年龄相关性白内障晶状体上皮细胞中的表达及其临床意义

目的:探讨血小板源性生长子A(Platelet derived growth factor-A,PDGF-A)在年龄相关性白内障晶状体上皮细胞中的表达及其临床意义。方法:以哈尔滨医科大学附属第四医院眼科2011年6月~2012年1月收治的72例年龄相关性白内障晶状体前囊膜作为实验组,10例无白内障发生的透明晶状体前囊膜作为正常对照组,采用SP免疫组织化学方法分别检测两组上皮细胞中PDGF-A的表达,运用图像分析软件进行量化,并采用SPSS11.0软件进行统计学分析探讨其与年龄相关性白内障临床特征之间的关系。结果:年龄相关性白内障及透明晶状体上皮细胞中均有PDGF-A表达,且在年龄相关性白内障中表达明显高于透明对照组,差异有统计学意义(P0.05)。随晶状体混浊程度加重,PDGF-A强阳性表达率逐渐升高,差异有统计学意义(P0.05);另外,不同年龄组白内障人群中PDGF-A表达无显著性差异(P0.05)。结论:①PDGF-A在年龄相关性白内障患者晶状体上皮细胞中呈高表达;②年龄相关性白内障患者中PDGF-A的表达量与晶状体的混浊程度呈正相关;③PDGF-A的过度表达可能在年轻人白内障的发生中起到了重要作用。     

糖尿病性白内障相关的酶及蛋白质

在我国白内障是造成视力障碍的主要因素。糖尿病性白内障(DC)是糖尿病的慢性并发症之一,其致盲率仅次于糖尿病视网膜病变(DR),糖尿病的发病率逐年上升的同时,DC的发病率也在增加。虽然白内障手术能够治愈DC,但研究人员仍致力于研究其发病机制以求通过药物途径治疗或预防DC。最近的研究显示,白内障的生成与晶状体内某些成分的改变有直接或间接的关系,DC发病过程中更是有一些特殊的改变:多元醇通路与DC的发展有着紧密的联系,有学者认为多元醇积聚诱发了白内障形成;氧化损伤在白内障形成过程中起了重要作用,而高血糖使得晶状体中多种抗氧化酶受损;晶状体本身是人体蛋白质含量最高的器官,白内障本质上即为结构蛋白的变性,而某些晶状体蛋白作为结构蛋白的同时又具有功能性蛋白的特性,其性质的改变引发晶状体混浊。本文针对DC相关的某些晶状体蛋白及酶类的研究进展做一综述。     

用二次谐波成像技术研究经飞秒激光切削后角膜变化

本文用二次谐波成像技术（second harmonic generation SHG）来研究飞秒激光切削后角膜结构的变化.在生物学研究,材料科学等方面都有很广泛应用的SHG成像技术能在不破坏的角膜情况下获得高对比度的角膜层析图像,分辨率为500 nm,实验装置是利用现有的双光子显微镜.本文还根据成像结果评价了飞秒激光在角膜切削中的质量,为飞秒激光微米级的精确切削和临床应用提供了实验支持.     

近视的药物治疗及手术治疗研究进展

近视是最常见的眼病之一,近年来,由于近视的发病率逐年上升,已成为世界性医学问题,近视眼的治疗也成为研究重点。药物研究和手术治疗是近视的治疗目前的研究热点,并已经取得一定得成果。药物方面,阿托品,托品酰胺,哌仑西平,消旋山莨菪碱对近视的控制作用都有被证实,但由于应用上还有一定的副作用,远期效果也不明确,需要进一步研究。手术方面,近视眼手术主要分为角膜屈光手术和眼内屈光手术。LASIK手术是目前应用最广泛的角膜屈光手术方法,SBK手术,飞秒激光手术等技术也日趋成熟。但是,角膜手术存在着一定的风险,应用范围也较为局限,需要进一步研究。眼内屈光手术主要为有晶体眼的人工晶体植入术(PIOL)和屈光性晶状体置换术,矫正的范围更加大,但是眼内手术并发症较多,也需要进一步研究和改进。     

PDGF-A在年龄相关性白内障晶状体上皮细胞中的表达及其临床意义

目的：探讨血小板源性生长子A（Platelet derived growth factor—A,PDGF-A）在年龄相关性白内障晶状体上皮细胞中的表达及其临床意义。方法：以哈尔滨医科大学附属第四医院眼科2011年6月-2012年1月收治的72例年龄相关性白内障晶状体前囊膜作为实验组,10例无白内障发生的透明晶状体前囊膜作为正常对照组,采用sP免疫组织化学方法分别检测两组上皮细胞中PDGF—A的表达,运用图像分析软件进行量化,并采用SPSSll．0软件进行统计学分析探讨其与年龄相关性白内障·临床特征之间的关系。结果：年龄相关性白内障及透明晶状体上皮细胞中均有PDGF—A表达,且在年龄相关性白内障中表达明显高于透明对照组,差异有统计学意义（P〈0．05）。随晶状体混浊程度加重,PDGF-A强阳性表达率逐渐升高,差异有统计学意义（P〈0．05）;另外,不同年龄组白内障人群中PDGF—A表达无显著性差异（P〉0．05）。结论：①PDGF—A在年龄相关性白内障患者晶状体上皮细胞中呈高表达;②年龄相关性白内障患者中PDGF—A的表达量与晶状体的混浊程度呈正相关;（3）PDGF．A的过度表达可能在年轻人白内障的发生中起到了重要作用。     

Expression of Cytokines,Chmokines and Growth Factors in Patients Undergoing Cataract Surgery with Femtosecond Laser Pretreatment

Purpose

To describe cytokines, chemokines and growth factors profiles in patients undergoing cataract surgery with femtosecond laser pretreatment and investigate their relationships with the postoperative in vivo inflammation index.

Methods

Aqueous humor was collected from 22 eyes after femtosecond laser pretreatment and from 22 eyes at the beginning of routine cataract surgery. The levels of 45 inflammation-related mediators were measured using multiplex fluorescent bead-based immunoassays. Laser flare photometry was measured preoperatively and at 1 day, 7 days and 30 days postoperatively.

Results

Compared with the control group, the femtosecond laser treatment group showed significantly higher aqueous humor levels of fibroblast growth factor (FGF-2), tumor necrosis factor (TNF)-α, leukemia inhibitor factor (LIF), interleukin (IL)-1ra and IL-18, and significantly lower aqueous humor levels of IL-9, platelet-derived growth factor (PDGF)-BB, eotaxin and TNF-β. Postoperative aqueous flare was significantly greater in the manual cataract surgery group at 1 day (p<0.001), 7days (p<0.001) and 30 days (p = 0.002).No correlation was found between the analyzed mediators and the aqueous flare values.

Conclusions

The expression profiles of cytokines, chemokines and growth factors and the correlations of these profiles with the in vivo inflammatory indexes for patients undergoing cataract surgery with femtosecond laser pretreatment were identified. Our data indicate a disturbance of postoperative inflammation response after femtosecond laser treatment.     

Non-Invasive Bleaching of the Human Lens by Femtosecond Laser Photolysis

Background

Globally, cataract is the leading cause of blindness and impaired vision. Cataract surgery is an attractive treatment option but it remains unavailable in sufficient quantity for the vast majority of the world population living in areas without access to specialized health care. Reducing blindness from cataract requires solutions that can be applied outside operating theatres. Cataract is a protein conformational disease characterized by accumulation of light absorbing, fluorescent and scattering protein aggregates. The aim of the study was to investigate whether these compounds were susceptible to photobleaching by a non-invasive procedure and whether this would lead to optical rejuvenation of the lens.

Methodology/Principal Findings

Nine human donor lenses were treated with an 800 nm infra-red femtosecond pulsed laser in a treatment zone measuring 1×1×0.52 mm. After laser treatment the age-induced yellow discoloration of the lens was markedly reduced and the transmission of light was increased corresponding to an optical rejuvenation of 3 to 7 years.

Conclusions/Significance

The results demonstrate that the age-induced yellowing of the human lens can be bleached by a non-invasive procedure based on femtosecond laser photolysis. Cataract is a disease associated with old age. At the current technological stage, lens aging is delayed but with a treatment covering the entire lens volume complete optical rejuvenation is expected. Thus, femtosecond photolysis has the potential clinical value of replacing invasive cataract surgery by a non-invasive treatment modality that can be placed in mobile units, thus breaking down many of the barriers impeding access to treatment in remote and poor regions of the world.     

Surgical applications of femtosecond lasers

Femtosecond laser ablation permits non‐invasive surgeries in the bulk of a sample with submicrometer resolution. We briefly review the history of optical surgery techniques and the experimental background of femtosecond laser ablation. Next, we present several clinical applications, including dental surgery and eye surgery. We then summarize research applications, encompassing cell and tissue studies, research on C. elegans, and studies in zebrafish. We conclude by discussing future trends of femtosecond laser systems and some possible application directions. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Laser nanosurgery in cell biology

Since their first use in the early 60's, pulsed lasers have become increasingly popular for their ability to ablate biological tissue. Short laser pulses allow high precision surgery for biological and medical applications with minimal invasiveness. Performing highly targeted manipulation and ablation allows experiments impossible so far in development biology, cellular biology or even assisted reproductive technologies and laser surgery has been increasingly used over the last five years to answer key questions in Biology. Recently, picosecond UV and femtosecond IR laser pulses have been used to cleave microtubules and to severe actin stress fibers in vivo with a spatial precision in the submicrometer range to study their dynamics without affecting cell viability. We review recent findings on the underlying principles of pulsed laser nanosurgery mechanisms showing how the use of ultra short laser pulses increases precision and non-invasiveness of laser surgery. We show how the understanding of the surgical process allows one to distinguish between single cell ablation in living organisms or intracellular nanosurgery in living cells and we review recent applications to the study of forces and the quantification of cytoskeleton dynamics.     

全飞秒激光与飞秒激光辅助Lasik治疗高度近视患者的效果及对角膜曲率、视觉质量的影响分析

摘要 目的：探究全飞秒激光与飞秒激光辅助Lasik治疗高度近视患者的效果,并分析治疗对患者角膜曲率、视觉质量的影响。方法：选择2020年7月至2022年7月在我院接受治疗的120例（240眼）高度近视患者为研究对象,按照随机数字表法结合患者意愿的方式将其区分为研究组（n=60,接受飞秒激光辅助Lasik治疗）与对照组（n=60,接受全飞秒激光治疗）,对比两组患者术前术后调制传递函数截止频率（MTF）值及客观散射指数（OSI）、角膜曲率变化、屈光度差异、视力情况差异,统计两组患者各类并发症发生率并进行比较。结果：（1）两组患者术前MTF值及OSI组间差异无统计学意义（P>0.05）,术后研究组患者的MTF值和OSI值均明显低于对照组,差异具有统计学意义（P<0.05）;（2）术前、术后90 d两组患者的最佳矫正视力组间差异无统计学意义（P>0.05）,但组内前后比较两组患者的最佳矫正视力均较术前有明显提高（P<0.05）;（3）术前两组患者的角膜前表面曲率以及后表面曲率组间差异均无统计学意义（P>0.05）,术后90 d时开展组间比较,研究组患者角膜前表面曲率中K1、K2以及Km值均明显低于对照组（P<0.05）,但后表面曲率中K1、K2以及Km值组间差异无统计学意义（P>0.05）;进一步分析显示,两组患者术前术后前表面曲率K1、K2以及Km值有明显变化（P<0.05）,后表面曲率K1、K2以及Km值前后差异无统计学意义（P>0.05）;（4）统计研究组患者共出现弥漫性层间反应2例,感染1例,角膜内生1例,并发症总发生率3.33%（4/120）,明显高于对照组的0.00%（0/120）（P<0.05）。结论：全飞秒激光和飞秒激光辅助Lasik术对高度近视均具有较好的治疗效果,相比于全飞秒激光术,飞秒激光辅助Lasik术在改善患者视觉质量方面明显占优,但其并发症发生率同样更高,全飞秒激光术术后视觉质量,但安全性更高,建议临床上结合患者实际情况灵活选择术式,以改善高度近视患者预后。     

Evaluation of a 345 nm Femtosecond Laser for Corneal Surgery with Respect to Intraocular Radiation Hazard

Purpose

We report our findings from a preclinical safety study designed to assess potential side effects of corneal ultraviolet femtosecond laser treatment on lens and retina.

Methods

Refractive lenticules (-5 dpt) with a diameter of 6 mm were created in the right cornea of eight Dutch Belted rabbits. Radiant exposure was 0.5 J/cm² in two animals and 18 J/cm² in six animals. The presence of lens opacities was assessed prior to and up to six months following laser application using Scheimpflug images (Pentacam, Oculus) and backscatter analysis (Opacity Lensmeter 702, Interzeag). Ganzfeld flash and flicker electroretinogram (ERG) recordings were obtained from both eyes prior to and up to six weeks following laser application. At the study endpoint, retinas were examined by light microscopy.

Results

Independent of energy dose applied, no cataract formation could be observed clinically or with either of the two objective methods used. No changes in ERG recordings over time and no difference between treated and untreated eye were detected. Histologically, retinal morphology was preserved and retinal pigment epithelium as well as photoreceptor inner and outer segments appeared undamaged. Quantitative digital image analysis did not reveal cell loss in inner or outer nuclear layers.

Conclusions

Our analysis confirms theoretical considerations suggesting that ultraviolet femtosecond laser treatment of the cornea is safe for intraocular tissues. Transmitted light including stray light induces no photochemical effects in lens or retina at energy levels much higher than required for the clinical purpose. These conclusions cannot be applied to eyes with pre-existing retinal damage, as these may be more vulnerable to light.     

In-vivo intratissue ablation by nanojoule near-infrared femtosecond laser pulses

Non-invasive intratissue ablation was performed in the cornea of living rabbits by using 80 MHz near-infrared intense nanojoule femtosecond laser pulses. The intratissue surgical effect was induced by multiphoton absorption at a wavelength of 800 nm and was ascertained by histological examination. Highly precise intratissue ablation was obtained with no detrimental effects to the overlying or underlying layers. Activated keratocytes in the laser-treated corneas were detected with two-photon imaging postoperatively. Intratissue femtosecond laser ablation thus has potential as a effective technique in refractive surgery for the treatment of visual disorders. This work was supported in part by the German Science Foundation.     

Investigations of Photosensitive Proteins by Serial Crystallography

This review contains recent data on serial femtosecond X-ray crystallography (SFX), based on a femtosecond X-ray free electron laser, as well as on the possibilities of its application for studying photosensitive proteins. Development of this method began rather recently, and it has already shown its effectiveness and some unique advantages over conventional X-ray structural analysis. This technology is especially promising for structural studies of membrane proteins and for kinetic studies. The main principle of the method, the possibility of its application in structural biology, its advantages and disadvantages, as well as its prospects for further development are analyzed in this review. Special attention is given to publications in which the SFX method has been used to study photosensitive proteins.