Pax6基因在视网膜母细胞瘤中的表达及临床意义

目的:探讨Pax6基因在视网膜母细胞瘤(retinoblastoma,Rb)中的表达及临床意义。方法:选择我院2001-01/2012-12收存在眼科病理室的15例Rb组织切片设为观察组,再选取15例正常视网膜组织切片设为对照组。应用Western-Blot及RT-PCR(逆转录酶链反应)法分别对正常视网膜组织及Rb组织中的Pax6蛋白和Pax6 mRNA的表达进行检测,同时应用Western-Blot法对Pax6基因下游的BRN3b及MATH5促分化基因在蛋白水平的表达进行检测,最后进行组间比较,进而对Pax6基因在Rb中的表达及临床意义进行探讨。结果:观察组Pax6基因mRNA表达平均值为0.99±0.03,Pax6基因蛋白表达平均值为2.07±0.15,BRN3b蛋白表达平均值为0.195±0.016,MATH5蛋白表达平均值为0.190±0.031,均明显高于对照组,差异有统计学意义(P<0.05)。结论:异常表达的Pax6基因可能对Rb的出现起到促进作用。     

HSP27、增殖细胞核抗原在视网膜母细胞瘤中的表达

目的探讨热休克蛋白(heat shock proteins,HSP)27、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在视网膜母细胞瘤(retinoblastoma,Rb)中的表达及其与Rb分化程度和视神经浸润的关系。方法应用鼠抗人HSP27及PCNA的单克隆抗体,采用PV9000二步法免疫组织化学方法,对36例Rb患儿和2例正常对照视网膜组织常规石蜡标本行HSP27及PCNA表达的测定,并分析它们与肿瘤分化程度及视神经浸润的关系。结果HSP27及PCNA在Rb中的表达阳性率分别为69.4%及83.3%;两指标在未分化组阳性表达高于分化组(P<0.05),两指标在视神经浸润组的阳性表达高于未浸润组(P<0.05)。Rb中HSP27的表达与PCNA的表达呈正相关(P<0.01)。结论Rb中HSP27和PCNA的表达与Rb的分化程度、视神经浸润有关。     

视网膜母细胞瘤增殖细胞核抗原表达的研究

目的 :探讨增殖细胞核抗原 (proliferatingcellnuclearantigen ,PCNA)在视网膜母细胞瘤 (retinoblastoma ,Rb)中的表达及其与Rb病理类型和侵犯视神经情况的关系。方法 :用免疫组化技术检测PCNA在 3 7例Rb组织标本中的表达 ,并应用多功能真彩色病理图像分析系统 (CMIAS系列 )来分析PCNA染色的阳性细胞个数 ,采用积分光密度值定量分析其增殖指数 ,并将各指标进行SPSS统计软件相关性分析。结果 :本组 3 7例Rb标本中PCNA的阳性表达为 97 3 0 % ,而在 2例正常视网膜组织内 (对照组 )的表达均为阴性。PCNA的表达水平与Rb不同的分化、增殖程度有显著的统计学差异 (P <0 0 1) ,Rb侵犯视神经情况越严重、分化程度越差 ,PCNA的表达越强。结论 :Rb中PCNA的高表达可作为判断Rb恶性及其程度的新型标记物。PCNA与Rb分化类型、视神经侵犯情况密切相关 ,为PCNA表达水平反映Rb分化程度和增殖能力提供理论依据     

热休克蛋白27及CD34在视网膜母细胞瘤的表达

目的 探讨热休克蛋白27（HSP27）、CD34在视网膜母细胞瘤（Rb）中的表达及其与Rb血管化、分化程度和视神经浸润的关系.方法 应用鼠抗人HSP27、CD34的单克隆抗体,采用北京中杉PV9000通用型二步法免疫组化法,对36例Rb常规石蜡标本行HSP27、CD34表达的测定,并计算CD34标记的肿瘤微血管密度（MVD）值.HSP27光密度值（OD值）、MVD在未分化组、分化组的差异及在有视神经浸润组、无视神经浸润组的差异采用t检验,HSP27 OD值与MVD之间的相关性采用Spearman等级相关分析.结果 HSP27在正常视网膜神经纤维层有弱表达,在Rb中的阳性表达率为69%,阳性物质主要位于细胞浆内.HSP27和CD34在Rb中的阳性表达率分别为69%和100%,CD34标记的MVD及HSP27均与Rb的分化程度、视神经浸润有关（P均＜0.01）.CD34标记的微血管密度与HSP27的表达呈正相关（r=0.4659,P＜0.01）.结论 HSP27、CD34标记的MVD的测定对于确定Rb的组织特性和肿瘤的生物学行为具有重要意义.HSP27可能会促进新生血管的发生,与肿瘤的浸润、转移有关.     

视网膜母细胞瘤多药耐药相关蛋白的研究

目的 检测P-糖蛋白(P-Gp)、多药耐药基因相关蛋白(MRP)及肺耐药相关蛋白(LRP)在视网膜母细胞瘤(Rb)中的表达及临床意义;初步分析Rb患者临床病理指标与MRP间的关系;探讨Rb多药耐药现象的可能机制.方法 实验研究.应用免疫组织化学染色方法检测P-gp、MRP、LRP在75例Rb肿瘤标本中的表达情况.分析3种蛋白表达的相关性及其与患者年龄、性别、眼别、临床表现、组织病理分化程度等临床病理指标的相关性.各蛋白表达情况与一般临床特点、组织病理学特征的比较采用卡方检验,各蛋白间的相关性采用多元相关分析.结果 P-Gp、LRP、MRP蛋自在Rb中阳性表达率分别为64.0%、25.3%、36.0%.P-gp与LRP、P-gp与MRP、LRP与MRP的共表达阳性率分别为18.7%、32.0%、20.0%.P-gp、LRP、MRP在分化型Rb组织中的阳性表达率均高于杀分化型,且组间差异具有统计学意义(χ2=8.002,χ2=17.327,χ2=28.421;P<0.05).3种蛋白的表达均与年龄、性别、眼别无关(χ2=0.003～3.385,P>0.05).P-gp、LRP的表达分别与MRP的表达其有相关性(r=0.389,r=0.521;P<0.05).结论 Rb原发性多药耐药的形成是一个多因素、多步弱的复杂过程,与包括P-gp、LRP、MRP等在内的多项因素的参与有关.P-gp、LRP、MRP蛋白可以作为反映Rb耐药的分子基础,耐药相关标志的联合检测更有利于准确判断Rb的多药耐药状态,为Rb化学治疗提供科学的理论依据.     

视网膜母细胞瘤血管生成拟态与临床病理学特征关系的研究

目的 探讨视网膜母细胞瘤(Rb)标本中是否存在血管生成拟态(VM),并分析VM与Rb临床病理特征及其与缺氧诱导因子-1αa(HIF-1α)、血管内皮生长因子(VEGF)蛋白表达的关系,进一步阐述VM的临床意义.方法 实验研究.收集60例Rb患者、10例正常人视网膜组织石蜡标本及相关临床病理资料,应用过碘酸-雪夫(PAS)与CD34双重染色以及免疫组织化学染色法检测60例Rb中是否存在VM和分布,观察与临床病理学特征的关系特点;检测HIF-1α、VEGF蛋白表达,分析Rb瘤组织中VM与HIF-1α、VEGF表达的关系;用CD34抗体进行肿瘤血管内皮染色,并计数肿瘤微血管密度(MVD).Rb和正常视网膜组织中的VM、HIF-1α、VEGF蛋白阳性表达的比较采用χ2检验;VM阳性组和VM阴性组的MVD值的比较采用q检验.结果 HE染色法观察到Rb中存在VM,其为Rb细胞构成的管腔样结构,无内皮细胞衬覆,腔内可见红细胞.CD34和PAS双重染色结合CD34、神经元特异性烯醇化酶(NSE)免疫组织化学染色进一步证明Rb中存在VM,60例Rb中VM阳性率为18.33%(11/60),其中随着R-E分级的增高,VM阳性表达率明显增加,差别具有统计学意义(χ2=8.861,P<0.05);分化型Rb中VM阳性率(4.34%)低于未分化型中VM阳性率(22.02%)(χ2=4.872,P<0.05);此外,HIF-1α、VEGF在Rb的VM形成过程中的阳性表达率依次为VM阳性组大于VM阴性组大于正常组;微血管密度计数高低与VM的表达有关,VM阴性组的MVD值(49.77±2.05)高于VM阳性组MVD值(36.53±1.15).结论 Rb中存在VM,且肿瘤恶性程度越高,形成VM能力越强.存在VM的Rb组织HIF-1α、VEGF高表达,提示HIF-1α、VEGF在VM的形成中可能起促进作用.     

视网膜母细胞瘤中EphB4和EphrinB2的表达及意义

目的 探讨EphB4和EphrinB2的蛋白在视网膜母细胞瘤中的表达及其意义.方法 应用免疫组织化学SP法测定10例正常视网膜组织和40例视网膜母细胞瘤标本中EphB4和EphrinB2的表达水平,分析其与性别、临床分期、分化程度和视神经浸润的关系.结果(1)EphB4和EphrinB2蛋白表达的阳性率,在正常视网膜组织中分别为5％和2.5％,40例视网膜母细胞瘤标本中分别为77.5％和67.5％.EphB4和EphrinB2在视网膜母细胞瘤组织中的阳性率与显著高于正常视网膜组织中的阳性率(P＜0.05).(2)EphB4和EphrinB2的蛋白在视网膜母细胞瘤的阳性表达率与肿瘤组织的临床分期、分化程度及视神经浸润显著相关(P＜0.05).(3)EphB4和EphrinB2的蛋白在视网膜母细胞瘤组织中的阳性表达显著相关(P＜0.05).结论 EphB4和EphrinB2可能在视网膜母细胞瘤的发生和发展过程中起重要作用,可作为评价视网膜母细胞瘤的浸润及预后的标志物.     

CD1a标记的肿瘤浸润性树突状细胞在视网膜母细胞瘤中的表达

目的:探讨浸润性树突状细胞(TIDC)在视网膜母细胞瘤(retinoblastoma,Rb)组织中的表达。方法:以鼠抗人CD1a单克隆抗体为一抗,采用免疫组化SP染色法检测31例Rb组织中树突状细胞的表达。结果:在所有31例标本中,有23例经免疫组化染色后有CD1a+细胞的表达,即阳性率为74.2%。其中高分化为主型阳性率为77.8%(7/9),未分化为主型阳性率为75%(15/20),自发退变型为50%(1/2)。结论:Rb组织中CD1a阳性细胞数量与肿瘤分型之间未见明显相关性,TIDC是否可看作一个良好的肿瘤预后指标,还有待进一步确证。     

视网膜母细胞瘤组织芯片中PTEN、mdm2、p53表达及与组织病理特征关系的研究

目的 探讨视网膜母细胞瘤组织芯片中PTEN,murine double minute 2(mdm2)及p53蛋白的表达及与肿瘤临床组织病理特征的关系.设计实验性研究.研究对象 64例视网膜母细胞瘤和6例正常视网膜组织.方法 免疫组化方法检测视网膜母细胞瘤组织芯片和正常视网膜组织石蜡标本中PTEN,mdm2及p53的表达情况,将肿瘤分为球后视神经受侵袭组和球后视神经未受侵组,检测两组间mdm2及p53表达差异,并结合肿瘤的组织病理学特征进行综合分析.主要指标视网膜母细胞瘤组织芯片中PTEN,mdm2和p53表达率.结果 视网膜母细胞瘤中PTEN,mdm2与p53的阳性表达率分别为53.13%,48.43%和53.13%.球后视神经受侵袭组较球后视神经未受侵组mdm2(P=0.027)及p53(P=0.017)表达均明显增高.视网膜母细胞瘤中PTEN与p53表达呈负相关(r=-0.384,P=0.045),mdm2与p53蛋白表达呈正相关(r=0.281,P=0.018).结论 视网膜母细胞瘤中mdm2和p53的过度表达与视神经侵袭程度存在一定相关性.mdm2及p53蛋白的检测可能成为视网膜母细胞瘤恶性程度的参考指标.     

视网膜母细胞瘤组织中DNA甲基转移酶1、DNA甲基转移酶3a、3b的表达

目的 观察视网膜母细胞瘤(RB)组织中DNA甲基转移酶(DNMT)1、DNMT3a和DN MT3b的表达.方法 62例RB肿瘤组织标本及6例正常视网膜组织标本纳入研究.其中,低分化组织标本17例,高分化组织标本45例；侵袭性肿瘤16例,非侵袭性肿瘤46例.采用免疫组织化学染色方法检测RB肿瘤组织标本中DNMT1、DNMT3a、DNMT3b的表达.以细胞核棕色染色为阳性表达,蓝色为阴性表达.以DNMT1、DNMT3a及DNMT3b阳性细胞分别≥65％、60％及40％为DNMT1、DNMT3a、DNMT3b高表达,≥1％但＜65％、60％及40％为低表达.同时分析DNMT1、DNMT3a、DNMT3b表达和MIB-1标记指数之间的相关性.结果 光学显微镜观察发现,正常视网膜组织中DNMT1、DNMT3a和DNMT3b均为阴性表达.肿瘤视网膜组织中均可见DNMT1、DNMT3a和DNMT3b表达,其阳性表达率分别为100％、98％、92％.高低分化组织标本比较,DNMT1、DNMT3a高表达率(x2=12.57、10.54)及阳性细胞计数(U=179、198)间差异均有统计学意义(P＜0.05)；DNMT3b高表达率(x2=1.5)和阳性细胞计数(U=307)间差异均无统计学意义(P＞0.05).侵袭性及非侵袭性肿瘤组织标本比较,DNMT1高表达率(x2=4.72)及阳性细胞计数(U=236)间差异均有统计学意义(P＜0.05)；DNMT3a、DNMT3b高表达率(x2=3.53、0.84)及阳性细胞计数(U=338、257)间差异均无统计学意义(P＞0.05).相关性分析结果显示,MIB-1标记指数与RB肿瘤组织中DNMT1、DNMT3a及DNMT3b表达呈正相关(R2=0.554、0.376、0.219,P＜0.05).结论 RB肿瘤视网膜组织中DNMT1、DNMT3a和DNMT3b均呈高表达.     

Paraneoplastic and non-paraneoplastic retinopathy and optic neuropathy: Evaluation and management

Paraneoplastic syndromes involving the visual system are a heterogeneous group of disorders occurring in the setting of systemic malignancy. Timely recognition of one of these entities can facilitate early detection and treatment of an unsuspected, underlying malignancy, sometimes months before it would have otherwise presented, and gives the patient an increased chance at survival. We outline the clinical features, pathogenesis, and treatment strategies for the retinal- and optic nerve–based paraneoplastic syndromes: cancer-associated retinopathy; melanoma-associated retinopathy; paraneoplastic vitelliform maculopathy; bilateral diffuse uveal melanocytic proliferation; paraneoplastic optic neuropathy; and polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes syndrome. Distinguishing these disorders from their non-paraneoplastic counterparts (e.g., autoimmune-related retinopathy and optic neuropathy, and acute zonal occult outer retinopathy) and determining appropriate systemic evaluation for the responsible tumor can be challenging. In addition, we discuss the utility and interpretation of autoantibody testing.     

Segmentation and accuracy in copying and drawing: Experts and beginners

As part of an ongoing investigation into real-world copying and drawing, I recorded the eye-hand drawing strategies of 16 subjects with drawing experiences ranging from expert to novice while they copied a line drawing of a standing nude. The experts produced accurate copies whereas all the beginners produced marked inaccuracies of overall scaling, proportion and shape. Analysis of eye and hand movements showed that the experts alone segmented the original drawing into simple line sections that were copied one at a time using a direct eye-hand strategy not requiring intermediary encoding to visual memory. The results suggest that segmentation into simple lines defines the task-specific process of accurate copying, and that this process is restricted to experts, i.e. acquired through training and practice. Additional preliminary tests also suggest that a similar process may apply to drawing a model from life.     

Nodes and nodal points and lines in eyes and other optical systems

The typical stigmatic optical system has two nodal points: an incident nodal point and an emergent nodal point. A ray through the incident nodal point emerges from the system through the emergent nodal point with its direction unchanged. In the presence of astigmatism nodal points are not possible in most cases. Instead there are structures, called nodes in this paper, of which nodal points are special cases. Because of astigmatism most eyes do not have nodal points a fact with obvious implications for concepts, such as the visual axis, which are based on nodal points. In order to gain insight into the issues this paper develops a general theory of nodes which holds for optical systems in general, including eyes, and makes particular allowance for astigmatism and relative decentration of refracting elements in the system. Key concepts are the incident and emergent nodal characteristics of the optical system. They are represented by 2 × 2 matrices whose eigenstructures define the nature and longitudinal position of the nodes. If a system's nodal characteristic is a scalar matrix then the node is a nodal point. Otherwise there are several possibilities: Firstly, a node may take the form of a single nodal line. Second, a node may consist of two separated nodal lines reminiscent of the familiar interval of Sturm although the nodal lines are not necessarily orthogonal. Third, a node may have no obvious nodal line or point. In the second and third of these classes one can define mid-nodal ellipses. Astigmatic systems exist with nodal points and stigmatic systems exist with no nodal points. The nodal centre may serve as an approximation for a nodal point if the node is not a point. Examples in the Appendix , including a model eye, illustrate the several possibilities.     

Sensitivity to first- and second-order motion and form in children and adults

We compared the sensitivity of adults and children aged 3-10 years to first- and second-order motion and form. For first-order stimuli, at all ages sensitivity was better for motion than form, and motion thresholds were better at 6 Hz than at 1.5 Hz. For second-order stimuli, at all ages sensitivity was better for form than motion, and motion thresholds were better at 0.25 cyc/deg than at 1 cyc/deg. Thresholds became adult-like later for motion than for form and later for first-order than second-order stimuli. For first-order stimuli, the changes with age were larger and more protracted.     

Ethics and optometry: concepts and applications

Ethics refers both to the study of behaviour, and moral principals. The related concepts of justice and law are also relevant to optometry. A profession typically claims specialist knowledge and ethical behaviour – putting the interests of clients above its own. However, professional codes fail as ethical directives, and their goals are questioned. Beginning with broad principles, institutional ethics and issues of general health care provision are considered, and applications to optometry are made. Ethical theory can guide us in interacting with our patients, utilising resources and ordering priorities. The conservative approach to consumerism and advertising is defended on the basis of protecting public and professional interests. Ethical behaviour can be fostered, and this process should begin in undergraduate education.     

Nutritional antioxidants and age-related cataract and maculopathy

Loss of vision is the second greatest, next to death, fear among the elderly. Age-related cataract (ARC) and maculopathy (ARM) are two major causes of blindness worldwide. There are several important reasons to study relationships between risk for ARC/ARM and nutrition: (1) because it is likely that the same nutritional practices that are associated with prolonged eye function will also be associated with delayed age-related compromises to other organs, and perhaps, aging in general, (2) surgical resources are insufficient to provide economic and safe surgeries for cataract and do not provide a cure for ARM, and (3) there will be considerable financial savings and improvements in quality of life if health rather than old age is extended, particularly given the rapidly growing elderly segment of our population. It is clear that oxidative stress is associated with compromises to the lens and retina. Recent literature indicates that antioxidants may ameliorate the risk for ARC and ARM. Given the association between oxidative damage and age-related eye debilities, it is not surprising that over 70 studies have attempted to relate antioxidant intake to risk for ARC and ARM. This article will review epidemiological literature about ARC and ARM with emphasis on roles for vitamins C and E and carotenoids. Since glycation and glycoxidation are major molecular insults which involve an oxidative stress component, we also review new literature that relates dietary carbohydrate intake to risk for ARC and ARM. To evaluate dietary effects as a whole, several studies have tried to relate dietary patterns to risk for ARC. We will also give some attention to this emerging research. While data from the observational studies generally support a protective role for antioxidants in foods or supplements, results from intervention trials are less encouraging with respect to limiting risk for ARC/ARM prevalence or progress through antioxidant supplementations, or maintaining higher levels of antioxidants either in diet or blood. Without more information it is difficult to parse these results. It would be worthwhile to determine why the various types of studies are not yielding similar results. However, there are many common insults and mechanistic compromises that are associated with aging, and proper nutrition early in life may address some of these compromises and provide for extended youthful function later in life. Indeed, proper nutrition, possibly including use of antioxidant supplements for the nutritionally impoverished, along with healthy life styles may provide the least costly and most practical means to delay ARC and ARM. Further studies should be devoted to identifying the most effective strategy to prevent or delay the development and progress of ARC/ARM. The efforts should include identifying the right nutrient(s), defining useful levels of the nutrient(s), and determining the age when the supplementation should begin.     

Exogenous attention and color perception: performance and appearance of saturation and hue

Exogenous covert attention is an automatic, transient form of attention that can be triggered by sudden changes in the periphery. Here we test for the effects of attention on color perception. We used the methodology developed by Carrasco, Ling, and Read [Carrasco, M., Ling, S., & Read, S. (2004). Attention alters appearance. Nature Neuroscience, 7 (3) 308-313] to explore the effects of exogenous attention on appearance of saturation (Experiment 1) and of hue (Experiment 2). We also tested orientation discrimination performance for single stimuli defined by saturation or hue (Experiment 3). The results indicate that attention increases apparent saturation, but does not change apparent hue, notwithstanding the fact that it improves orientation discrimination for both saturation and hue stimuli.     

血糖和血脂异常代谢与糖尿病性眼肌麻痹患者的关联分析

目的探究糖代谢和脂代谢异常与糖尿病性眼肌麻痹患者的关联性。方法选取2012年2月至2016年2月在首都医科大学附属北京同仁医院神经内科住院的39例糖尿病性眼肌麻痹患者为观察组,40例于本院体检中心进行健康检查的人群为对照组。对比两组研究对象糖代谢(Hb A1c、FPG、2h PBG)及脂代谢(TG、TC、HDL-C、LDL-C)水平;分析观察组患者脂代谢指标中TG、TC与糖代谢指标Hb A1c的相关性。结果观察组糖代谢的Hb A1c、FPG及2h PBG水平显著高于对照组;观察组脂代谢的TG、TC及LDL-C水平显著高于对照组,指标HDL-C水平低于对照组;观察组血糖血脂的关联性分析得出TG及TC与Hb A1c呈正相关,均处于平行上升趋势。结论糖尿病患者高血糖和血脂代谢异常是引发颅神经病变的因素,导致患者眼肌麻痹并发症的出现,治疗和改善糖尿病患者血糖及血脂代谢异常,可以更好的预防糖尿病患者眼肌麻痹的发生。