增生型糖尿病视网膜病变患者外周血内皮祖细胞的数量变化及意义

Objective To investigate the amounts of endothelial progenitor cells(EPCs)in peripheral blood of patients with proliferative diabetic retinopathy(PDR).Methods Forty patients with PDR(PDR group),thirty patmnts with type 2 diabetes mellitus(DM)without DR(DM group),and twenty agematched normal subjects(control group)were enrolled in this study.Blood samples were treated bv repeated centrifugation and stained with monoclonal antibodies.At least 2 × 105 cells were analyzed bv flow cytometry.EPCs were identified by CD34 and CD133 antibody.The correlation between EPCs numbers and DR duration,glycosylated hemoglobin,serum lipids was analyzed.Results The number of EPCs in PDR,DM and control group were(49±12)、(35±11)、(90±25)cells/ml respectively,the difference was statistically significant(F=56.260,P=0.000).There was a positive correlation between EPCs numbers and DR duration(r=0.564,P＜0.05).However there was no correlation between EPCs numbers and glycosylated hemoglobin(r=-0.170,P>0.05)or triglyceride levels(r=0.261,P>0.05).Conclusions The number of EPCs in peripheral blood of PDR patients was decreased. EPCs might play an important role in the pathogenesis of PDR.     

增生型糖尿病视网膜病变患者外周血内皮祖细胞的数量变化及意义

Objective To investigate the amounts of endothelial progenitor cells(EPCs)in peripheral blood of patients with proliferative diabetic retinopathy(PDR).Methods Forty patients with PDR(PDR group),thirty patmnts with type 2 diabetes mellitus(DM)without DR(DM group),and twenty agematched normal subjects(control group)were enrolled in this study.Blood samples were treated bv repeated centrifugation and stained with monoclonal antibodies.At least 2 × 105 cells were analyzed bv flow cytometry.EPCs were identified by CD34 and CD133 antibody.The correlation between EPCs numbers and DR duration,glycosylated hemoglobin,serum lipids was analyzed.Results The number of EPCs in PDR,DM and control group were(49±12)、(35±11)、(90±25)cells/ml respectively,the difference was statistically significant(F=56.260,P=0.000).There was a positive correlation between EPCs numbers and DR duration(r=0.564,P＜0.05).However there was no correlation between EPCs numbers and glycosylated hemoglobin(r=-0.170,P>0.05)or triglyceride levels(r=0.261,P>0.05).Conclusions The number of EPCs in peripheral blood of PDR patients was decreased. EPCs might play an important role in the pathogenesis of PDR.     

红细胞增多症影响早产儿视网膜病变的临床观察

Objective To explore the effect of polycythemia on retinopathy of prematurity (ROP).Methods The clinical data of 262 premature cases was analyzed retrospectively in Xi'an Children Hospital from January 2005 to January 2009. Polycythemia was found in 46 cases (17. 56 %), including 27 males and 19 females. In 216 infants without polycythemia (82.46%), 155 were male and 61 were female. The difference of the birth weight (t = 0. 730, P = 0. 466), gestational age (t= 1. 603, P= 0. 110), oxygen inhalation numbers (χ2 =0. 04 ,P>0. 90) and times (t= 1. 225, P= 0. 223), and concentration (t= 1. 823,P=0. 071) between polycythemia group and no polycythemia group were not significant. In order to diagnose ROP, the ocular fundus of all premature infants was examined with binocular indirect ophthalmoscope, and the stage of ROP was assessed. Results In all the premature infants, ROP was found in 120 cases (45.80%). In 46 cases of polycythemia, ROP was found in 25 cases (54.34%); in 216 infants without polycythemia, ROP was found in 95 cases (43.98%); the difference of ROP incidence betweenthe two groups was not significant (χ2=1.64, P>0. 1). In 120 ROP patients, 104 cases (86.67%) with ROP< 3 stage and 16 cases (13.33%) with ROP ≥3 stage were found. In 25 ROP patients with polycythemia, 18 cases (72.00%) with ROP <3 stage and 7 cases (28. 00%) with ROP ≥3 stage were found. In 95 ROP patients without polycythemia, 86 cases (90. 53%) with ROP <3 stage and 9 cases (9. 47%) with ROP ≥3 stage were found. The difference of the incidence of ROP 0. 05). Conclusion Polycythemia may not affect the incidence of ROP, but impact on the severity of ROP.     

Frequency of IL-10-producing regulatory B cells associated with disease activity in thyroid-associated orbitopathy

AIM: To investigate the association between IL-10-producing regulatory B (B10) cells and the clinical features of thyroid-associated orbitopathy (TAO). METHODS: A total of 30 patients with TAO were recruited at Zhongshan Ophthalmic Center from May 2015 to December 2015. Peripheral blood mononuclear cells (PBMCs) were separated from blood samples of 30 TAO patients and 16 healthy controls and stimulated with CD40 ligand and CpG for 48h. The frequency of IL-10+ B cells was examined by flow cytometry and the correlation between the frequency of IL-10+ B cells and clinical features of TAO was analyzed by SPSS. RESULTS: The frequency of IL-10+ B cells among CD19+ B cells in TAO patients was significantly lower than in healthy controls (TAO: 4.66%±1.88% vs healthy control: 6.82%±2.40%, P<0.01). The frequency of IL-10+ B cells showed a positive correlation with disease activity of TAO measured by Clinical Activity Score (CAS) (r=0.50, P<0.01), and became higher in TAO patients with family history of Graves’ disease (GD) (P=0.04). CONCLUSION: The decrease of the frequency of IL-10+ B cells in TAO patients indicates the deficiency of B10 cells in TAO, and the positive association with disease activity suggests its important role in TAO inflammation regulation.     

红细胞增多症影响早产儿视网膜病变的临床观察

Objective To explore the effect of polycythemia on retinopathy of prematurity (ROP).Methods The clinical data of 262 premature cases was analyzed retrospectively in Xi'an Children Hospital from January 2005 to January 2009. Polycythemia was found in 46 cases (17. 56 %), including 27 males and 19 females. In 216 infants without polycythemia (82.46%), 155 were male and 61 were female. The difference of the birth weight (t = 0. 730, P = 0. 466), gestational age (t= 1. 603, P= 0. 110), oxygen inhalation numbers (χ2 =0. 04 ,P>0. 90) and times (t= 1. 225, P= 0. 223), and concentration (t= 1. 823,P=0. 071) between polycythemia group and no polycythemia group were not significant. In order to diagnose ROP, the ocular fundus of all premature infants was examined with binocular indirect ophthalmoscope, and the stage of ROP was assessed. Results In all the premature infants, ROP was found in 120 cases (45.80%). In 46 cases of polycythemia, ROP was found in 25 cases (54.34%); in 216 infants without polycythemia, ROP was found in 95 cases (43.98%); the difference of ROP incidence betweenthe two groups was not significant (χ2=1.64, P>0. 1). In 120 ROP patients, 104 cases (86.67%) with ROP< 3 stage and 16 cases (13.33%) with ROP ≥3 stage were found. In 25 ROP patients with polycythemia, 18 cases (72.00%) with ROP <3 stage and 7 cases (28. 00%) with ROP ≥3 stage were found. In 95 ROP patients without polycythemia, 86 cases (90. 53%) with ROP <3 stage and 9 cases (9. 47%) with ROP ≥3 stage were found. The difference of the incidence of ROP 0. 05). Conclusion Polycythemia may not affect the incidence of ROP, but impact on the severity of ROP.     

人角膜内皮细胞压力调节培养的初步研究

Objective To investigate the effect of pressure bionic culture on the morphology and function of rabbit corneal endothelial cells. Methods Corneal endothelial cells were separated and purified by tearing apart the descemet and digesting with trypsin and EDTA, then cultured in the plate. The cells were divided into two groups: group A were cultured under atmosphere; cells exposed to 2 kPa( 14. 66 mm Hg) pressure in vitro was group B; the morphology and growth pattern of cells were observed by inverted microscope; cells origin were identified by neuron-specific enolase immunoassay. Cellular changes in the structure were observed by HE staining and scanning and transmission electron microscopy (SEM and TEM) analysis. Cells activity was detected by flow cytometry. Results NSE antibody of the primary corneal endothelial cells was positive without corneal epithelial cells and corneal stroma cells. Two groups of cells were cultured for 120-144 h respectively, the morphology was flat, polygon, most of cells were hexagon and abundant cytoplasms in group B (pressure bionic culture), but in group A, the cells size was not uniform and there were much granules in the cytoplasm. There was no difference in the time of formation of monolayer in two groups. SEM showed that cells exposed to pressure connected tightly and the surface was rich in microvilli, extended foot processes and attached to the substrate tightly, while cells cultured under atmosphere with more off-chip. In group B, Annexiv-FITC/PI detection of apoptosis showed cell survival rate was 98.2%, early apoptosis rate was 0.7%, late apoptosis rate was 1.0%, death rate was 0. 1%; the corresponding data were 92.2%, 5.2%, 2.3%, and 0.3% in group A, respectively; There was statistically significant difference between the two groups (x2 =594. 0,P ＜0. 01 ). After cultured for 96 h,the expression of ZO-1 protein in cells exposed to pressure was higher than those in control. Conclusions The biological activity of endothelial cells is regulated positively by bionic pressure. The establishment of a new biomimetic pressure model will help to investigate the physiological function and injury repair of corneal endothelial cells in vitro.     

Anti-scarring effect of sodium hyaluronate at filtration pathway after filtering surgery in rabbits

AIM: To investigate the anti-scarring effect of sodium hyaluronate (HA) at filtration pathway after filtering surgery in a rabbit model. METHODS: Fifteen healthy adult New Zealand white rabbits were selected for trabeculectomy in both eyes. The right eyes were used as HA group with 0.1 mL HA injected into the anterior chamber at the end of the operation; the left eyes were used with 0.1 mL sodium lactate Ringer’s solution (RS) injected into the anterior chamber as RS group. Intraocular pressure (IOP), filtering blebs morphology, inflammatory reaction and complications were observed at the 7, 60, and 90d after surgery. RESULTS: One day after surgery, the IOP of HA and RS groups were 12.75±1.92 and 10.50±1.59 mm Hg (P=0.005). At the 7th day postoperative, the filtering blebs of each group were functional type and TGF-β expression was significantly difference in both groups (0.10±0.01 vs 0.14±0.02, P=0.024). After 60d of the operation, all filtering blebs were scarring and alpha-smooth muscle actin (α-SMA) expression was significantly difference in both groups (0.40±0.04 vs 0.35±0.02, P=0.032). α-SMA positive cells were mainly distributed in the junction of conjunctiva and sclera and around the blood vessels. The collagen volume fraction (CVF) of HA and RS group was (75.49±7.01)% and (79.93±5.35)% (P=0.044). On the 90th day after the operation, CVF was (82.57±5.19)% and (88.08±1.75)% in HA and RS groups (P=0.036). There was no α-SMA positive cell in HA group, while a few positive cells were observed in RS group (P=0.000). CONCLUSION: HA has effect of anti-scar and anti-inflammation on filtration pathway after filtering surgery within 3mo by inhibiting fibroblast proliferation and collagen deposition.     

早产儿视网膜病变激光光凝治疗后视网膜功能发育状况观察

Objective To observe the development of cone/rod functions in children with retinopathy of prematurity(ROP)after laser photocoagulation.Methods 30 ROP patients(60 eyes)treated with laser photocoagulation(case group)and 30 preterm infants(60 eyes)without ROP(control group)were enrolled in this study.Flash Electroretinogram(F-ERG)was performed on all the children, and the response of rod cells and cone cells,maximal mixed responses and oscillatory potentials were recorded.Results Compared with the control group,the amplitude of response of rod cells was obviously decreased(t=-2.385,P＜0.05),while the latency phase of rod cells was obviously prolonged(t=-2.799,P＜0.05);the amplitudes of a-and b-wave of maximal response were significantly decreased in the case group(t=-2.967.-4.037;P＜0.05).But there was no significant difference of amplitudes(t=-1.402)and latency phase (t=-1.637,0.465)0f b-/a-wave of the maximal mixed response between two groups(P>0.05).In the response of cone cells,there was no significant difference of the latency phase(t=1.222)and amplitude (t=-0.636)of a-wave as well as amplitude(t=-1.927)of b-wave between two groups(P>0.05).The latency phase of b-wave of cone cells in the case group was longer than that in the control group(t=-2.466,P＜0.05).Conclusions Compared to no-ROP preterm infant retina,laser-treated ROP retina has normal cone function development and delayed rod function development.     

早产儿视网膜病变激光光凝治疗后视网膜功能发育状况观察

Objective To observe the development of cone/rod functions in children with retinopathy of prematurity(ROP)after laser photocoagulation.Methods 30 ROP patients(60 eyes)treated with laser photocoagulation(case group)and 30 preterm infants(60 eyes)without ROP(control group)were enrolled in this study.Flash Electroretinogram(F-ERG)was performed on all the children, and the response of rod cells and cone cells,maximal mixed responses and oscillatory potentials were recorded.Results Compared with the control group,the amplitude of response of rod cells was obviously decreased(t=-2.385,P＜0.05),while the latency phase of rod cells was obviously prolonged(t=-2.799,P＜0.05);the amplitudes of a-and b-wave of maximal response were significantly decreased in the case group(t=-2.967.-4.037;P＜0.05).But there was no significant difference of amplitudes(t=-1.402)and latency phase (t=-1.637,0.465)0f b-/a-wave of the maximal mixed response between two groups(P>0.05).In the response of cone cells,there was no significant difference of the latency phase(t=1.222)and amplitude (t=-0.636)of a-wave as well as amplitude(t=-1.927)of b-wave between two groups(P>0.05).The latency phase of b-wave of cone cells in the case group was longer than that in the control group(t=-2.466,P＜0.05).Conclusions Compared to no-ROP preterm infant retina,laser-treated ROP retina has normal cone function development and delayed rod function development.     

增生型糖尿病视网膜病变患者外周血内皮祖细胞的数量变化及意义

目的 观察增生型糖尿病视网膜病变(PDR)患者外周血内皮祖细胞(EPCs)数量的变化规律,探讨EPCs在糖尿病视网膜病变(DR)发病机制中的作用.方法 PDR患者40例(PDR组)、单纯2型糖尿病患者30例(糖尿病组)及健康体检者20名(对照组)纳入研究.抽取3组受检者晨起空腹静脉血2 ml,反复离心过滤.采用流式细胞仪分析2×105个细胞,以CD34和(或)CD133双阳性细胞群为EPCs,观察并比较3组受检者外周血EPCs数量变化;同时分析PDR患者外周血EPCs数量与DR病程、糖化血红蛋白及血脂的相关性.结果 PDR组、糖尿病组及对照组受检者外周血EPCs计数分别为(49±12)、(35±11)、(90±25)个/ml,3组间差异有统计学意义(F=56.260,P＜0.05).相关性分析发现,PDR患者外周血EPCs数量与DR病程呈正相关(r=0.564,P＜0.05),与糖化血红蛋白(r=-0.170,P>0.05)及血脂(r=0.261,P>0.05)均无相关性.结论 PDR患者外周血EPCs数量较正常者明显降低;EPCs在DR发病机制中可能具有一定作用.

Abstract：

Objective To investigate the amounts of endothelial progenitor cells(EPCs)in peripheral blood of patients with proliferative diabetic retinopathy(PDR).Methods Forty patients with PDR(PDR group),thirty patmnts with type 2 diabetes mellitus(DM)without DR(DM group),and twenty agematched normal subjects(control group)were enrolled in this study.Blood samples were treated bv repeated centrifugation and stained with monoclonal antibodies.At least 2 × 105 cells were analyzed bv flow cytometry.EPCs were identified by CD34 and CD133 antibody.The correlation between EPCs numbers and DR duration,glycosylated hemoglobin,serum lipids was analyzed.Results The number of EPCs in PDR,DM and control group were(49±12)、(35±11)、(90±25)cells/ml respectively,the difference was statistically significant(F=56.260,P=0.000).There was a positive correlation between EPCs numbers and DR duration(r=0.564,P＜0.05).However there was no correlation between EPCs numbers and glycosylated hemoglobin(r=-0.170,P>0.05)or triglyceride levels(r=0.261,P>0.05).Conclusions The number of EPCs in peripheral blood of PDR patients was decreased. EPCs might play an important role in the pathogenesis of PDR.     

眼眶腺样囊性癌的治疗与预后分析

目的 分析总结眼眶腺样囊性癌的治疗方法及影响预后的因素,为改善预后提供帮助.方法 采用回顾性系列病例研究,分析1991年1月至2006年7月就诊的75例眼眶腺样囊性癌患者的手术记录、病理分型及随访记录.相关数据采用χ2检验和Fisher精确检验进行统计分析.结果 眼眶实体型腺样囊性癌的2年复发率为85%(17/20)、5年复发率为100%(19/19),而腺样.管状型则分别为23.53%(8/34)和64.52%(20/31),差异有统计学意义(2年,χ2=19.14,P=0.000;5年,Fisher精确检验,P=0.003).前者发生局部蔓延和远处转移例数亦多于后者.肿瘤切除术后放射治疗的5年复发率为70%(14/20),低于单纯手术切除的复发率92.86%(13/14)(Fisher精确检验,P=0.198).首次手术行眶内容物剜除术的5年复发率为25%(1/4),低于复发后再行眶内容物剜除术的病例为75%(6/8)(Fisher精确检验,P=0.222),γ刀、粒子刀、化疗及生物治疗的效果不能确定.局部蔓延主要是至颅内、副鼻窦和颞窝,远处转移可到达肺、骨、肝、耳前淋巴结.5年远处转移率为25.71%(9/35),肺转移和骨转移各占33.33%(3/9).5年生存率74.29%(26/35),死亡率25.71%(9/35),无瘤生存率37.14%(13/35),10年无瘤生存率17.14%(6/35).最常见的死亡原因是颅内蔓延.肿瘤切除联合放射治疗可以使5年生存率提高到80%(16/20).结论 腺样囊性癌是高度恶性的眼眶肿瘤,复发率和死亡率均较高,病理分型、治疗方法均影响预后.采取综合治疗方法,可以减少复发,提高生存率.     

CD44V6蛋白在眼部鳞状细胞癌中的表达及与增殖细胞核抗原关系的研究

目的探讨眼部鳞状细胞癌中细胞黏附分子变异体6(cluster of differentiation 44 variant 6,CD44V6)蛋白表达及与增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的关系.方法应用链霉亲和素-生物素-过氧化物酶复合物免疫组化法检测35例眼部鳞状细胞癌、20例乳头状瘤和11例正常眼睑皮肤组织标本CD44V6蛋白表达.结果 CD44V6蛋白在眼部鳞状细胞癌(squamous cell carcinoma,SCC)中的阳性表达率为62.9%(22/35),而在乳头状瘤的阳性表达率为15.0%(3/20),在11例正常眼睑皮肤组织中均无CD44V6蛋白的阳性表达;恶性组CD44V6蛋白的阳性表达率明显高于良性组(χ2=11.757,P＜0.01)和正常组(P= 0.000 1),8例淋巴转移的CD44V6蛋白的阳性表达率明显高于无转移者(P=0.049);CD44V6蛋白的阳性表达组其PCNA指数明显高于阴性表达组,差异有非常显著性(t=20.21,P＜0.01).结论 CD44V6分子可能与眼部鳞状细胞癌的恶性表型、浸润及转移有关,CD44V6与PCNA蛋白的阳性表达呈正相关,两者同时检测可能对判定患者预后有临床参考价值.     

泪腺肿瘤中细胞黏附分子CD44变异体6及纤维连接蛋白的表达与预后相关性探讨

目的　探讨细胞黏附分子CD44变异体 6 (CD44V6)和纤维连接蛋白 (FN)的表达与泪腺肿瘤发生、发展和预后的关系及其临床病理学意义。方法　应用链霉亲和素 生物素 过氧化物酶复合物免疫组化法检测 4 9例泪腺良恶性肿瘤组织和 8例正常泪腺组织的CD44V6和FN蛋白的表达。并对检测结果进行比较。结果　CD44V6、FN蛋白在良性混合腺瘤的阳性表达率分别为 30 8% (8/ 2 6 )和80 8% (2 1/ 2 6 ) ,在泪腺恶性肿瘤的阳性表达率分别为 6 0 9% (14 / 2 3)和 34 8% (8/ 2 3) ,经统计学分析 ,恶性组的阳性表达率与良性组的阳性表达率差异有显著意义 (P <0 0 5 ) ;在复发肿瘤标本中 ,CD44V6、FN蛋白的阳性表达率与无复发组的阳性表达率比较 ,差异有显著意义 (P <0 0 5 ) ;两者表达呈显著负相关性 (P <0 0 1)。结论　CD44V6、FN在泪腺肿瘤的发生、发展及复发过程中发挥重要作用 ,且 2者的表达呈负相关性 ,检测CD44V6、FN蛋白的表达有助于了解泪腺肿瘤生物学行为及对患者预后的判断     

WERI-Rb1肿瘤干细胞的分离扩增及鉴定

目的　分离培养人视网膜母细胞瘤细胞系ＷＥＲＩ-Ｒｂ１中的肿瘤干细胞,建立肿瘤干细胞的鉴定方法。方法　将ＷＥＲＩ-Ｒｂ１细胞系接种至无血清培养基,对其中的肿瘤干细胞行分离、培养扩增及传代,观察细胞形态,采用旁群细胞检测、克隆形成实验、ＣＤ１３３流式细胞仪检测、荧光定量ＰＣＲ等方法对肿瘤干细胞特性进行鉴定。结果　体外扩增培养的ＷＥＲＩ-Ｒｂ１细胞系呈疏松葡萄状团块,ＷＥＲＩ-Ｒｂ１细胞系中存在比例稳定的旁群细胞（０．０７５±０．０１７）％,无血清培养基可分离扩增培养出ＷＥＲＩ-Ｒｂ１中的肿瘤干细胞,形成类似胚胎干细胞的克隆团块,后者具备较高的克隆形成能力,克隆形成率为（３１．７０±１．８９）％,且干细胞标记物ＣＤ１３３的基因及蛋白均高表达,ＣＤ１３３基因表达量为ＷＥＲＩ-Ｒｂ１的（２．２５±０．１９）倍。结论　本研究阐明了视网膜母细胞瘤细胞系ＷＥＲＩ-Ｒｂ１肿瘤干细胞的分离、培养、扩增及鉴定方法,证明了ＷＥＲＩ-Ｒｂ１中肿瘤干细胞的存在,为后续对视网膜母细胞瘤的治疗提供线索,为进一步研究视网膜母细胞瘤肿瘤干细胞及胚胎干细胞等的特性对比提供细胞来源。     

两种绷带镜在糖尿病患者玻璃体视网膜手术中应用后对角膜的影响

目的:观察不同类型的角膜绷带镜在玻璃体视网膜手术中应用后对糖尿病患者角膜的影响.方法:前瞻性非随机对照研究.将符合增殖性糖尿病视网膜病变、拟行玻璃体视网膜手术的患者69例69眼按照患者意愿分为两组.A组36例36眼,术中使用直径为13.8 mm角膜绷带镜覆盖于角膜表面,非接触广角镜下实施玻璃体切割联合眼内填充术,手术结束后去除绷带镜;B组33例33眼,同法使用直径为14.0 mm角膜绷带镜,以相同方式实施手术.术前和术后1、2、3、7d进行裂隙灯显微镜、角膜荧光素钠染色检查.术前和术后7d行角膜内皮细胞计数、角膜厚度测量.结果:术前两组患者角膜荧光素钠染色阳性检出率分别为42%和42%(x2=0.004,P=0.949).术后第1、2、3、7d两组患者角膜染色阳性率分别为47%和45%(x2=0.022,P=0.883),44%和45%(x2=0.007,P=0.933),44%和42%(x2=0.029,P=0.886),42%和39%(x2=0.037,P=0.848);两组患者间各时间点角膜上皮阳性检出率比较,差异无统计学意义(P>0.05).术前和术后7d时,A组角膜内皮细胞计数分别为2779.25±329.55、2777.14±331.17个/mm2,差异无统计学意义(t=0.551,P=0.585);B组角膜内皮细胞计数分别为2678.61±335.64、2672.45±336.25个/mm2,差异无统计学意义(t=1.774,P=0.086).术前和术后7 d时A组角膜厚度分别为519.25±23.42、542.03±25.94μm,差异有统计学意义(t=-6.854,P<0.001);B组为525.64±20.97、551.33±27.87μm,差异有统计学意义(t=-7.204,P<0.001).结论:糖尿病患者玻璃体视网膜手术中使用不同类型角膜绷带镜,手术前后角膜上皮完整性无差异.不同类型的绷带镜均可有效保护角膜上皮,术中可保持良好的角膜透明性.     

Existence of small slow-cycling Langerhans cells in the limbal basal epithelium that express ABCG2

Despite the obvious importance of limbal stem cells in corneal homeostasis and tumorigenesis, little is known about their specific biological characteristics. The purpose of this study was to characterize limbal slow-cycling cells based on the expression of ABCG2 and major histocompatibility complex (MHC) class II and the cell size. Wistar rats were daily injected with 5-bromo-2-deoxyuridine (BrdU) at a dose of 5 mg/100 g for 2 weeks. After 4-week BrdU-free period, corneal tissues were excised, and immunofluorescence staining for ABCG2, BrdU, and MHC class II was performed by confocal microscopy. In another series, corneal tissues of normal rat were double immunostained for ABCG2, keratin 14, keratin 3, CD11c, and MHC class II. In addition, limbal, peripheral and central corneal epithelial sheets were isolated by Dispase II digestion and dissociated into single cell by trypsin digestion and cytospin preparations were double immunostained for ABCG2 and MHC class II. The cell size and nucleus-to-cytoplasm (N/C) ratio of limbal ABCG2+ cells were analyzed and compared with those of cells from other zones. BrdU label-retaining cells (LRCs) with expression of ABCG2 were found in the limbal epithelial basal layer, but not in other parts of the cornea. Approximately 20% of these cells were MHC class II positive. All MHC class II+ cells in the corneal epithelium were positive for CD11c, a marker for dendritic cells (DCs). Double labeling with ABCG2 and keratin 14 showed that nearly four-fifth of limbal ABCG2+ cells were positive for keratin 14 but negative for keratin 3, exhibiting an undifferentiated epithelial cell lineage. Cytospin sample analysis revealed the presence of a distinct population of smaller ABCG2+ cells with expression of MHC class II with a larger N/C ratio in the limbal epithelium. A new population of small slow-cycling cells with large N/C ratio has been found to express ABCG2 in the limbal epithelial basal layer. Some of these cells normally express MHC class II antigen. These findings may have important implications for our understanding of the characteristics of limbal slow-cycling cells.     

Corneal parameters and their correlations with refractive error,axial length,anterior chamber depth and lens thickness in black South Africans

目的:收集南非健康黑人的角膜直径(CD)、曲率(ACC)和角膜中央厚度(CCT)的数据并探讨其与屈光不正等效球镜(SE)、眼轴长度(AL)、前房深度(ACD)、晶状体厚度(LT)的相关性,及三个角膜参数之间的相关性.方法:选取600例受试者,包括305例男性和295例女性(平均年龄:28.15±13.1y,年龄范围:10~66y)接受全面的眼科检查,包括验光,裂隙灯和眼底镜检查.Oculus Keratograph 4检测CD和ACC,ivue-100光学相干断层扫描检测CCT.Nidek AR-310 A自动验光仪和主观验光检测屈光不正.Nidek US-500 A超检测AL,ACD和LT.结果:由于左右眼参数评估的差异无统计学意义,此处只显示右眼的结果.角膜各项参数平均值如下:CD=11.77±0.32 mm(10.30-13.70 mm),ACC=7.88±0.29 mm(7.13~8.88 mm),CCT=493.05±33.2μm(412~590μm).CD与SE无相关性(r=0.05,P=0.24).CD与AL(r=0.58,P=0.00)、ACD(r=0.63,P=0.00)显著相关,但与LT(r=-0.40,P=0.00)呈负相关.ACC与SE(r=-0.03,P=0.48)无显著相关性.ACC与AL(r=0.40,P=0.00)呈正相关.ACC与ACD(r=0.04,P=0.56)、LT(r=-0.03,P=0.88)无显著相关性.CCT与角膜以外其他参数间无显著相关性.角膜各项参数的相关性表明,CD与ACC(r=0.71,P=0.00)、CCT(r=-0.68,P=0.00)相关.结论:CD与AL、ACD、LT相关,ACC与AL相关.CCT与角膜以外其它参数无相关性,是一个独立的因素.在角膜各项参数中,CD与ACC、CCT相关.     

获得性免疫缺陷综合征患者视网膜微血管病变与巨细胞病毒视网膜炎对比研究

目的　分析获得性免疫缺陷综合征（acquired immunodeficiency syndrome，AIDS）患者视网膜微血管病变与巨细胞病毒视网膜炎（cytomegalovirus retinitis，CMVR）的疾病特征。方法　回顾性病例分析。选取就诊于首都医科大学附属北京佑安医院眼科并经感染科专科医师确诊的AIDS患者145例，其中合并视网膜微血管病变者75例为微血管病变组，合并CMVR者70例为CMVR组。所有患者均进行AIDS相关病史询问，同时行血人类免疫缺陷病毒（HIV）载量和CD4+T细胞检测，并行最佳矫正视力、眼压、裂隙灯检查、散瞳眼底检查及彩色眼底照相等眼科检查。对全身情况允许的微血管病变组25例、CMVR组70例进行血液和房水CMV-DNA检测。结果　视网膜微血管病变组最佳矫正视力为0.91±0.16，13例(17.33%)患者已开始高效联合抗反转录病毒治疗（HAART），CD4+T细胞数为22（0～506）个·μL^-1，8例（10.67%）患者有眼前节反应；CMVR组患者最佳矫正视力为0.63±0.37，34例(48.57%)患者已开始HAART，CD4+T细胞数为13（1~48）个·μL^-1，23例(32.85%)患者有眼前节反应，上述指标两组相比差异均有统计学意义（均为P=0.000）。微血管病变组25例中房水CMV-DNA阳性0例，血液CMV-DNA阳性8例（32.00%）；CMVR组70例中房水CMV-DNA阳性60例（85.71%），血液CMV-DNA阳性33例（47.14%）；两组血液中CMV-DNA阳性率相比差异无统计学意义（P=0.227），但房水中阳性率两组相比差异有统计学意义（P=0.000）。结论　AIDS合并CMVR患者比AIDS合并视网膜微血管病变患者视力更差，CD4+T细胞计数更低，眼前节反应及房水CMV-DNA阳性率更高，且视网膜微血管病变患者未接受HAART的人数明显高于CMVR患者。     

内毒素诱导的大鼠葡萄膜炎的巨噬细胞中Toll样受体4的表达

目的 探讨在内毒素诱导的Wistar大鼠葡萄膜炎中Toll样受体4(TLR4)阳性细胞与虹膜组织中巨噬细胞的动态变化和分布.方法 实验研究.Wistar大鼠50只,用随机数字法随机分为5组,每组10只,分别为正常对照(0 h)组、6 h组、12 h组、24 h组及48 h组.除0 h组外其余各组均足垫部注射霍乱弧菌内毒素200μg,注射后于裂隙灯显微镜下观察双眼前节炎症反应变化.按实验分组于0、6、12、24、48 h处死大鼠.取虹膜一睫状体及脉络膜组织.通过葡萄膜铺片免疫组织化学方法检测TLR4和巨噬细胞的标记CD163的表达.人工计数虹膜中TLR4~+与CD163~+的细胞并计算细胞密度,计算圆形和多形性的CD163~+细胞占所有CD163~+细胞的百分比.进一步采用免疫荧光双标记检测TLR4和CD163共表达的情况.通过单因素方差分析分别对大鼠虹膜内阳性细胞密度以及圆形、多形性CD163~+细胞的百分比进行统计学检验.结果 正常大鼠虹膜睫状体组织不表达TLR4.6 h组有2只大鼠虹膜内可见少量TLR4~+细胞,12～48 h组所有大鼠虹膜内TLR4~+细胞明显增多(F=167.2,P＜0.001),虹膜内TLR4~+细胞密度分别为(506.1±39.5)个/mm~2(12 h组)、(492.3±54.5)个/mm~2(24 h组)及(663.8±150.2)个/mm~2 (48 h组).在注射LPS后12～48 h期间TLR4~+细胞形态无明显变化.0～48 h组大鼠虹膜内均有CD163~+细胞,0 h组圆形和多形性CD163~+细胞百分比为13%,12～48 h组其百分比约为80%,且圆形细胞主要位于虹膜基质层.免疫荧光双标记可见TLR4和CD163的共表达,TLR4位于细胞膜,CD163位于细胞质.5组大鼠脉络膜内均未见TLR4表达.结论 内毒素诱导的大鼠葡萄膜炎中虹膜内TLR4表达增高,部分虹膜固有巨噬细胞表达TLR4.TLR4可能在葡萄膜炎的发生发展中起一定作用.     

泪腺腺样囊性癌高级别转化的临床病理学研究

目的探讨泪腺腺样囊性癌高级别转化(LACC-HGT)的临床病理学特征。 方法回顾性分析首都医科大学附属北京同仁医院病理科2011年1月至2019年12月间确诊的8例LACC-HGT病例。其中，男性4例(4只眼)，女性4例(4只眼)；年龄19~65岁，中位年龄44.5岁。记录所有患者的性别、年龄、首发症状、肿瘤大小及骨质破坏情况；观察组织病理学特点；利用免疫组织化学EnVision法标记P63、腺泡来源标志物转录因子SRY相关蛋白10(SOX10)、细胞角蛋白7(CK7)、分化簇117(CD117)、Ki-67、原癌基因MYB、人表皮生长因子受体-2(Her-2)及雄激素受体(AR)等的表达情况。 结果8例LACC-HGT患者中，临床表现为眼球突出者5例，占62.5%(5/8)；眼睑肿胀者1例，占12.5%(1/8)；眼球突出伴眼睑肿胀者2例，占25%(2/8)。肿瘤大小2~5 cm，平均(3.50±0.92)cm。骨质破坏者6例，占75%(6/8)。2例随访数据不全，其余6例随访期间均出现转移，转移部位包括腮腺、肺、脑、骨及区域淋巴结等，其中4例死亡。患者存活时间3~54个月，中位生存时间26个月。8例肿物均为实性，肿物最大直径2~5 cm，平均3.5 cm。肿物切面灰白、实性且质地中等，与周围组织边界不清。8例LACC-HGT病例中均可见经典ACC区域。其中，低级别的筛状型结构者2例，占25%(2/8)；实性型结构者3例，占37.5%(3/8)；筛状与实性混合型且实性成分>30%者3例，占37.5%(3/8)。高级别转化(HGT)区域占肿瘤镜下总面积的比例为60%~90%，肿瘤细胞排列呈实性未分化癌或低分化腺癌形态，片状；实性片巢常大于一个低倍镜视野。细胞异型性明显，胞核显著增大，核仁及核分裂象易见(平均38.75个/10高倍镜视野)。病理组织学显示神经浸润者8例，占100%(8/8)；脉管侵犯者5例，占62.5%(5/8)；间质促纤维结缔组织反应者7例，占87.5%(7/8)；坏死者6例，占75%(6/8)，其中4例伴钙化。免疫组织化学染色显示在经典腺样囊性癌(ACC)区域，P63在管周、筛周及筛孔周的肌上皮细胞核表达，在实性巢中可呈斑驳状表达；CK7和CD117在管腔面腺上皮以及筛状结构中向导管分化的细胞表达，而在实性巢中表达较弥漫；MYB蛋白在肌上皮和腺上皮中均呈胞核表达；在HGT区域，P63阳性肿瘤细胞明显减少，部分区域完全消失，而CK7和CD117在肿瘤中呈胞膜弥漫强阳性表达，提示腺泡来源的标志物SOX10呈胞核阳性表达，MYB蛋白亦呈核阳性表达。Ki-67增殖指数在HGT区域约为40%~70%(平均52.5%)，在经典ACC区域约为10%~30%(平均15%)。Her-2和AR在ACC及HGT区域均呈阴性表达。 结论LACC-HGT组织病理学特征显著，生物学行为差，病理医师应根据组织病理学及免疫组织化学特点做出正确诊断，以指导临床治疗及预后评估。