应用免疫荧光技术对眼瘢痕性类天疱疮的研究

目的 探讨免疫荧光技术对眼瘢痕性类天孢疮（ＯＣＰ）的实验诊断意义。方法 应用直接免疫荧光（ＤＩＦ）检测１６例ＯＣＰ患者的结膜和应用间接免疫荧光（ＨＦ）以鼠食道和正常人结膜为基质检测３４例ＯＣＰ患者的血清。结果 ＤＩＦ证实６例结膜上皮细胞基底膜上有免疫复合物（ＩｇＧ和／或ＩｇＡ）和／或Ｃ３呈线状沉积。以结膜为基质的ＨＦ证实血清中抗基底膜（ＢＭＺ）抗体４例,抗细胞间物质（ＨＳ）抗体１４例,总阳性率为５     

角膜共聚焦显微镜在眼表疾病诊治中的作用

角膜共聚焦显微镜是从细胞水平上对活体角膜进行无创观察的新型高分辨率显微镜。通过对全层角膜组织的三维、实时和活体条件下的光学断层扫描成像,在角膜组织的生理、病理研究和眼表疾病的诊断和治疗中体现出极大的优越性。（跟科,2011．20：145-147）     

Eosinophil granule proteins expressed in ocular cicatricial pemphigoid

BACKGROUND—Blister formation and tissue damage in bullous pemphigoid have been attributed to the release of eosinophil granule proteins—namely, to eosinophil derived cationic protein (ECP) and major basic protein (MBP). In the present investigation these eosinophil granule proteins were studied in the conjunctiva of patients with ocular cicatricial pemphigoid (OCP).
METHODS—Conjunctival biopsy specimens obtained from patients with subacute (n=8) or chronic conjunctival disease (n=13) were analysed histologically and immunohistochemically using antibodies directed against EG1 (stored and secreted ECP), EG2 (secreted ECP), MBP, CD45 (common leucocyte antigen), CD3 (pan T cell marker), and HLA-DR (class II antigen).
RESULTS—Subepithelial mononuclear cells, mast cells, and neutrophils were detected in all specimens. The number of mononuclear cells, neutrophils, CD45+ cells, CD3+ cells, and the HLA-DR expression were significantly higher in the subacute than in the chronic disease group. Some eosinophils were found in specimens from five of eight patients with subacute OCP, but in none of the patients with chronic disease. The eosinophil granule proteins (ECP and MBP) were found in the epithelium and substantia propria in patients with subacute conjunctivitis.
CONCLUSIONS—Subepithelial cell infiltration in the conjunctiva greatly differs between subacute and chronic ocular cicatricial pemphigoid specimens. The findings suggest that eosinophil granule proteins may participate in tissue damage in acute phase of inflammation in OCP.

     

眼瘢痕性类天疱疮机制及治疗研究进展

眼瘢痕性类天疱疮(ocular cicatricial pemphigoid,OCP)是黏膜类天疱疮(mucous membrane pemphigoid,MMP)在眼部的一种特殊表现形式,目前发病机制尚不明确,可由抗原抗体反应、炎细胞浸润、细胞因子作用、钙离子水平升高、易感基因等多种因素导致。病变早期表现为慢性进行性的结膜纤维化性炎症,晚期可见角膜混浊及新生血管形成,最终导致视力严重丧失。因此,及时对OCP患者进行规范的临床治疗尤为重要。例如氨苯砜、免疫球蛋白(intravenous immunoglobulin,IVIG)、利妥昔单抗(rituximab,RTX)、肿瘤坏死因子拮抗剂、肾上腺皮质激素类药物,可以有效控制炎症反应、延缓疾病进展。手术治疗可在OCP患者并发严重倒睫、角膜病变及白内障时酌情考虑。     

Clinical corneal confocal microscopy

Confocal microscopy allows non-invasive in vivo imaging of the ocular surface. Its unique physical properties enable microscopic examination of all layers of the cornea and have been used to investigate numerous corneal diseases: epithelial changes, numerous stromal degenerative or dystrophic diseases, endothelial pathologies, corneal deposits, infections, and traumatic lesions. It offers a new approach to study the physiological reactions of the cornea to different stimuli and the pathophysiologic events leading to corneal dysfunction in certain diseases. Confocal microscopy proves to be a powerful diagnostic tool and is especially of value in certain corneal diseases by allowing straightforward and non-invasive recognition of the pathologic conditions.     

瘢痕性类天疱疮角膜病变手术治疗

目的 观察眼瘢痕性类天疱疮(OCP)行羊膜移植和穿透性角膜移植术(PKP)后的效果.设计 回顾性病例系列.研究对象 OCP患者3例(4眼).方法 2例(3眼)患者为三级睑球粘连,术前视力HM/5cm～HM/10cm,行睑球粘连分离联合羊膜移植,术后放置绷带性角膜接触镜;另外1例(1眼)因并发角膜溃疡伴穿孔,行PKP,供体为甘油冷冻保存的角膜,观察术后角膜、结膜愈合情况.主要指标 视力,角膜、结膜愈合情况.结果 羊膜移植2例(3眼)患者术后1个月羊膜逐步吸收,2例(2眼)手术1年后视力0.04～0.05,中央角膜上皮化良好,二级睑球粘连,角膜缘新生血管减少.PKP后3周植片融解严重,1个月后行全角膜移植联合眼前节切除,再次发生植片融解,眼内容物流失而眼球萎缩.结论 部分眼表功能不全的OCP患者睑球粘连分离联合羊膜移植对减轻睑球粘连、提高视力有一定意义;由于多种不利因素影响,穿透性角膜移植治疗OCP并发穿孔性角膜溃疡效果不良.     

Clinical and microstructural analysis of patients with hyper-reflective corneal endothelial nuclei imaged by in vivo confocal microscopy

The purpose of this study was to determine the significance of hyper-reflective corneal endothelial nuclei imaged by in vivo confocal microscopy. A retrospective analysis was performed using a database of 505 patients that had undergone in vivo confocal microscopy of the cornea. All subjects with hyper-reflective endothelial nuclei were identified and these images were analysed to determine corneal endothelial cell density and morphology. The clinical notes of these patients were reviewed and corresponding data regarding corneal thickness was obtained from a related database of Orbscan II pachymetry. Hyper-reflective endothelial nuclei were identified in 41 eyes of 39 (7.7%) patients. Diagnoses included previous cataract surgery or penetrating keratoplasty, posterior polymorphous dystrophy, Fuchs' endothelial dystrophy and irido-corneal endothelial syndrome. No patients with clinically normal corneas exhibited bright endothelial nuclei. The mean endothelial cell density in this group was 1325+/-872 cells mm(-2) and endothelial density was below age-adjusted normal values in 69.2% of patients. Both cellular polymegathism (coefficient of variation of cell area 33.9+/-7.4%) and cellular pleomorphism were noted (51.8+/-9.0% hexagonal cells). The mean central corneal thickness was 582+/-52 microm. There was no significant difference in endothelial density and morphology compared to cases that had low endothelial density but did not exhibit bright nuclei. In conclusion, this study is the first to investigate the significance of bright endothelial nuclei detected by in vivo confocal microscopy. The strong association with corneal disease states suggests that the most likely explanation for this appearance is the alteration in cellular/nuclear morphology, composition or function.     

共聚焦显微镜在活体角膜上皮肿瘤鉴别诊断中的应用

目的 通过活体共聚焦显微镜在角膜上皮肿瘤诊断中的应用,分析、鉴别角膜上皮肿瘤性质.方法 7例角膜上皮肿瘤患者术前均行常规眼部裂隙灯检查、眼前节OCT和活体共聚焦显微镜检查.所有患者均实施角膜肿瘤切除术,术后行肿瘤组织病理学检查,与术前共聚焦显微镜结果比较.结果 7例角膜上皮肿瘤均侵犯透明角膜、角膜缘和球结膜,突起于眼球表面,形状似桑葚,血管丰富.在眼前节OCT检查中5例角膜肿瘤显示密度均匀,与角膜组织之间界限清晰,浅层基质中未见肿瘤密度影;另外2例角膜肿瘤显示,密度不均,与角膜组织界限不清,而且侵犯前弹力层和浅层角膜基质.在活体共聚焦显微镜中7例均显示角膜上皮细胞非典型增生,其中2例非典型增生明显,侵犯前弹力层,在角膜上皮层和浅基质层内形成“癌巢”,5例诊断为角、结膜乳头状瘤,2例诊断为角膜鳞状上皮细胞癌,与术后组织病理学诊断结果一致.结论 活体共聚焦显微镜检查对角膜上皮肿瘤术前诊断肿瘤性质提供重要依据.     

The clinical value of in vivo confocal microscopy for diagnosis of ocular surface squamous neoplasia

Purpose

To determine the reliability and efficiency of in vivo confocal microscopy for the diagnosis of ocular surface squamous neoplasia (OSSN).

Methods

A case series with five consecutive cases of OSSN were investigated retrospectively, of which the characteristics and subspecial types had been estimated by in vivo confocal microscopy before surgery. The structure and cellular features of OSSN were analyzed with other examinations, such as anterior-segment optical coherence tomography (AS-OCT), and confirmed by histopathological biopsy.

Results

The tumors revealed red gelatinous surfaces with vascular dilatation on the ocular surface of the conjunctival and corneal epithelium in anterior segment photography. Involvement of only corneal epithelium was observed by AS-OCT in three cases, whereas the Bowman''s layer and anterior stroma were also invaded in the other two cases. In vivo confocal microscopy showed cellular anisocytosis and enlarged nuclei with high nuclear to cytoplasmic ratio in three cases diagnosed as conjunctival intraepithelial neoplasia; moreover, nests were partially formed by isolated keratinized, binucleated, and actively mitotic dysmorphic epithelial cells in the other two cases diagnosed as carcinoma in situ and ocular surface squamous carcinoma (OSSC). The characteristics assessed from histopathological biopsy were similar to that revealed by in vivo confocal microscopy in all five cases.

Conclusion

In vivo confocal microscopy analysis of cytological characteristics of OSSN is a safe, relatively noninvasive, and effective diagnostic tool in detecting characteristics of OSSN before surgical resection. Although in vivo confocal microscopy cannot replace excisional biopsy for definitive diagnosis, it can be valuable for initial diagnosis and management of patients with OSSN.     

In vivo confocal microscopy in a patient with conjunctival amyloidosis

Conjunctival amyloidosis is a rare clinical entity. We herein reported in vivo confocal microscopic features of conjunctival amyloidosis and correlated the results with the histopathological findings. Histopathological examination of the biopsy specimen revealed acellular, amorphous, eosinophilic material in the substantia propria of the conjunctiva and positive staining with Congo red confirmed that the material was amyloid. Confocal microscopy with Heidelberg Retina Tomograph II Rostock Cornea Module showed hyporeflective material deposited in a lobular pattern in the substantia propria and around the blood vessels. The material seemed acellular and no inflammatory or dendriform cell accumulation was noted within or around the lesion. Further studies should be done to understand the confocal characteristics of the conjunctival diseases and find specific clues for the diagnosis.     

Conjunctival mucoepidermoid carcinoma in a patient with ocular cicatricial pemphigoid and a review of the literature

Invasive mucoepidermoid carcinoma of the conjunctiva of the left lower eyelid was diagnosed in an orbital exenteration specimen of a 57-year-old woman, after a biopsy of the same lesion was originally diagnosed as invasive squamous cell carcinoma. The woman was undergoing mitomycin C injections for ocular cicatricial pemphigoid, diagnosed in the same eye 2 years prior to identification of the neoplasm. The tumor invaded the cornea, sclera, lacrimal gland, regional small nerves, and lymphatics, but did not show intraocular involvement. The original biopsy specimen was reassessed with stains for mucin and found to be mucoepidermoid carcinoma of the conjunctiva. We reviewed 21 cases of mucoepidermoid carcinoma of the conjunctiva described to date in the English literature. We believe this number underestimates the true incidence of this condition, as it is frequently misdiagnosed both clinically and histopathologically. Evaluating suspected aggressive squamous cell carcinoma with special stains for mucin generally helps to identify mucoepidermoid carcinoma of the conjunctiva. More extensive surgical excision than that used for squamous cell carcinoma should be implemented in the management of mucoepidermoid carcinoma of the conjunctiva to prevent recurrence.     

瘢痕性天疱疮35例临床分析

目的:探讨瘢痕性天疱疮临床特点,为临床诊治提供帮助。方法:回顾性分析2002-01/2008-01我科诊治的35例瘢痕性天疱疮临床特点。结果:瘢痕性天疱疮平均发病年龄为62.4岁,男女性别比为20∶15,病史平均3.3a,51.43%以口腔黏膜病变首发,31.43%以眼部病变首发,眼睑内翻倒睫/慢性结膜炎和干眼/角膜病变为5/4/2,17.14%以其它黏膜病变首发。经过治疗,71.43%(25/35)患者症状消失或缓解超过3mo,14.29%(5/35)患者症状反复或无明显缓解,14.29%(5/35)患者中途退出治疗。治疗过程中37.14%(13/35)患者出现全身其它异常,其中6例出现肝功能异常,2例终止治疗,肾功能异常2例,1例终止治疗,血液系统异常3例,1例退出治疗,心肌梗塞1例,退出治疗,明显血压升高1例。结论:瘢痕性天疱疮是一种慢性病,多发于老年人,男性稍多于女性,眼组织经常受累,多表现为干眼症、睑球粘连和内翻倒睫。瘢痕性天疱疮治疗复杂,经常需要多学科协作。     

In vivo corneal confocal microscopic analysis in patients with keratoconus

AIM:To quantify corneal ultrastructure using laser scanning in vivo confocal microscopy (IVCM) in patients with keratoconus and control subjects.METHODS: Unscarred corneas of 78 keratoconic subjects without a history of contact lens use and 36 age-matched control subjects were evaluated with slit-lamp examination (SLE), corneal topography and laser scanning IVCM. One eye was randomly chosen for analysis. Anterior and posterior stromal keratocyte, endothelial cell and basal epithelial cell densities and sub-basal nerve structure were evaluated.RESULTS: IVCM qualitatively demonstrated enlarged basal epithelial cells, structural changes in sub-basal and stromal nerve fibers, abnormal stromal keratocytes and keratocyte nuclei, and pleomorphism and enlargement of endothelial cells. Compared with control subjects, significant reductions in basal epithelial cell density (5817±306 cells/mm² vs 4802±508 cells/mm², P<0.001), anterior stromal keratocyte density (800±111 cells/mm² vs 555±115 cells/mm², P<0.001), posterior stromal keratocyte density (333±34 cells/mm² vs 270±47 cells/mm², P<0.001), endothelial cell density (2875±223 cells/mm² vs 2686±265 cells/mm², P<0.001), sub-basal nerve fiber density (31.2±8.4 nerves/mm² vs 18.1±9.2 nerves/mm², P<0.001), sub-basal nerve fiber length (21.4±3.4 mm/mm² vs 16.1±5.1 mm/mm², P<0.001), and sub-basal nerve branch density (median 50.0 (first quartile 31.2 - third quartile 68.7) nerve branches/mm² vs median 25.0 (first quartile 6.2 - third quartile 45.3) nerve branches/mm², P<0.001) were observed in patients with keratoconus.CONCLUSION: Significant microstructural abnormalities were identified in all corneal layers in the eyes of subjects with keratoconus using IVCM. This non-invasive in vivo technique provides an important means to define and follow progress of microstructural changes in patients with keratoconus.     

In vivo confocal microscopy of climatic droplet keratopathy

We describe the corneal microstructural changes in a patient with spheroidal degeneration using in vivo confocal microscopy. Multiple hypo‐ and hyper‐reflective spherical lesions were observed in the anterior corneal stroma and Bowman's layer ranging from 45 to 220 μm in size. The corneal epithelium, posterior stroma and endothelium were otherwise unaffected. In vivo confocal microscopy demonstrates good correlation with excised histological samples in climatic droplet keratopathy. It provides a non‐invasive technique to examine the living cornea for degenerative disease and acts as a bridge between clinical and laboratory observations.     

Automatic analysis of corneal nerves imaged using in vivo confocal microscopy

Interest has grown over the past decade in using in vivo confocal microscopy to analyse the morphology of corneal nerves and their changes over time. Advances in computational modelling techniques have been applied to automate the estimation of sub‐basal nerve structure. These objective methods have the potential to quantify nerve density (and length), tortuosity, variations in nerve thickness, as well as temporal changes in nerve fibres such as migration patterns. Different approaches to automated nerve analysis, methods proposed and how they were validated in previous literature are reviewed. Improved understanding of these approaches and their limitations will help improve the diagnostic leverage of emerging developments for monitoring the onset and progression of a broad class of systemic diseases, including diabetes.     

Incidence, prevalence, and demographic characteristics of ocular cicatricial pemphigoid in Colombia: data from the National Health Registry 2009-2019

AIM: To provide an epidemiological characterization of ocular cicatricial pemphigoid (OCP) in Colombia. METHODS: We conducted a cross-sectional study using SISPRO. We applied the specific code of the International Classification of Diseases for Ocular Pemphigoid, from 2009-2019 to estimate prevalence, incidence, and the demographic status of the disease in Colombia. RESULTS: The estimated average prevalence was 0.22 per 1 000 000 inhabitants, and the estimated average incidence was 0.24 per 1 000 000 inhabitants. With a female predominance of 62.5%, and a male/female ratio of 1:1.6. The group of patients diagnosed with the disease after the age of 80 presented the highest prevalence. The departments with the highest prevalence were Antioquia, Bogotá, and Santander. CONCLUSION: There are important differences between worldwide and Colombian prevalence and incidence data, which may be related to genetic and epigenetic factors, and the possible underdiagnosis of the disease. According to the results, OCP is an extremely rare disease in Colombia. Nevertheless, it is important to encourage awareness of the disease due to its devastating consequences.     

Imaging posterior polymorphous corneal dystrophy by in vivo confocal microscopy

To identify features of posterior polymorphous dystrophy (PPMD) by in vivo confocal microscopy, the corneas of a female patient with PPMD were examined using slit‐lamp biomicroscopy and slit‐scanning in vivo confocal microscopy. Characteristic endothelial vesicular and band lesions were seen clinically and easily identified using in vivo confocal microscopy. However, endothelial pleomorphism, an increased density and reflectance of posterior stromal keratocytes, and prominence of corneal nerves were also delineated. In vivo confocal microscopy enhances clinicopathological diagnosis and follow up of corneal dystrophies with subtle clinical presentations, such as PPMD.     

The significance of Vogt's striae in keratoconus as evaluated by in vivo confocal microscopy

Background: To evaluate the association of the presence, extent and width of Vogt's striae with other microstructural corneal alterations in keratoconus using in vivo confocal microscopy (IVCM). Methods: Sixty‐eight keratoconic corneas of 68 patients were evaluated with slit‐lamp examination (SLE), corneal topography and IVCM. For each eye, the presence, extent and width of alternating light and dark bands (Vogt's striae) observed using IVCM was recorded together with keratocyte and endothelial cell densities, stromal nerve thickness, subbasal nerve density and thickness. The refractive status and the mean and steepest corneal curvatures were noted. Results: Vogt's striae were present in 43 (63.2%) eyes on SLE and dark bands were present in 53 (77.9%) eyes on IVCM. Compared with patients without dark bands, patients with dark bands had significantly higher refractive errors in spherical equivalents (SE; ?8.15 ± 3.70 vs. ?5.18 ± 2.46 diopters [D], P = 0.007), higher astigmatic errors (?5.88 ± 2.69 vs. ?4.10 ± 1.84 D, P = 0.027), higher steepest corneal curvatures (54.33 ± 4.38 vs. 51.23 ± 3.72 D, P = 0.018), lower anterior stromal keratocyte densities (1106 ± 172 vs. 1222 ± 171 cells/mm², P = 0.022) and lower nerve fibre densities (18.74 ± 6.54 vs. 22.66 ± 6.47 nerves/mm², P = 0.054). Compared with patients in whom dark bands were confined to the posterior stroma, patients with dark bands extending into the anterior stroma had significantly higher refractive errors in SE (?11.17 ± 2.25 vs. ?6.34 ± 3.48 D, P < 0.001), higher astigmatic errors (?7.44 ± 2.56 vs. ?4.69 ± 2.22 D, P = 0.006) and wider bands (6.0 ± 2.1 vs. 9.6 ± 3.1 µm, P < 0.001). Conclusions: Vogt's striae appear to be more prevalent in keratoconic corneas than can be appreciated clinically. The presence of Vogt's striae may be associated with corneal topographic and microstructural changes.     

In vivo confocal microscopy of pre-endothelial deposits

BACKGROUND: Deposits in various layers of the cornea might result from long-term medical therapy, photorefractive surgery, and longterm use of contact lenses or corneal dystrophies. METHODS: A 46-year-old woman was referred to our department with the suspected diagnosis of posterior polymorphous dystrophy. Slit-lamp biomicroscopy revealed bilateral small-sized deposits in the posterior part of the cornea. In vivo confocal microscopy was performed to evaluate these deposits in detail. RESULTS: In vivo confocal microscopy of the cornea identified hyperreflec-tive "dot-like" structures in the deep stromal layer and anterior to the endothelial cell layer. The morphology and number of keratocytes of the posterior stroma and of endothelial cells appeared normal. CONCLUSIONS: In vivo confocal microscopy is a very useful tool to analyze and visualize pre-endothelial deposits. Because there is no family history of corneal disease, the exact origin of the pre-endothelial deposits in our case remains unclear.