流式细胞术在眼部疾病细胞因子和细胞学检查中的应用进展

流式细胞术是一种能够对单个细胞或生物颗粒进行多参数、定量分析和分选的检测技术,是近年来迅速发展起来的一项先进的细胞定量分析技术,广泛应用于免疫学、肿瘤学和血液学等多领域医学研究和临床诊断。该技术可以定量检测和分析眼内液中细胞因子水平及各细胞亚群的分布特征。与传统的酶联免疫吸附试验、细胞形态学检查及免疫组织化学检测等方法比较,流式细胞术具有操作简单、所需样本量小、灵敏度高、通量高等优点。流式细胞术已广泛应用于眼内新生血管相关疾病、眼内淋巴瘤、视网膜病变、白内障、葡萄膜炎、结节病性葡萄膜炎、感染性眼内炎症性疾病等眼部疾病相关细胞因子水平检测和细胞亚群分析中,对眼部疾病的发病机制研究和靶向治疗起到日益重要的作用。本文就流式细胞术在眼部疾病眼内液的细胞因子和细胞学检查中的应用进展进行综述。     

腺病毒性角结膜炎的病原学诊断技术

腺病毒性角结膜炎是一种常见的眼部病毒感染性疾病,严重时可以暴发性流行.诊断主要依靠其临床表现,常存在误诊、漏诊,因此应重视其病原学诊断.国内外常用的病原学诊断技术包括病毒分离培养、免疫荧光技术、酶免疫技术、核酸杂交技术、PCR技术、循环抗体的检测等.本文对这些腺病毒性角结膜炎病原学诊断技术的病理、特点及其临床应用价值进行总结.     

细胞因子检测方法研究进展及其在眼内液检测中的应用

细胞因子是指由细胞分泌的、能调节细胞功能的小分子多肽,其在免疫调节、应答过程中发挥着重要作用。细胞因子检测方法主要包括生物活性检测法、免疫学检测法和分子生物学检测法。各种检测方法各有优势,不可相互替代。眼内液是指眼球中的房水及玻璃体液,与血液相比,眼内液细胞因子检测能更好地反映眼内的环境。近几年,细胞因子研究增多,检测方法也不断推陈出新。故本文中笔者拟对细胞因子的检测现状、进展及其在眼内液检测中的应用做一综述。     

庆大霉素在眼科的应用及其毒性作用

庆大霉素在眼科的应用及其毒性作用夏翠然综述刘英奇审校（河北医学院第三附属医院石家庄０５００１１）近几年来，庆大霉素已在眼科临床广泛应用，它可以预防和治疗各种细菌引起的急慢性眼部感染性疾病，它在眼内渗透性良好，能在炎症眼内迅速获得有效治疗浓度［１］。为...     

35例葡萄膜炎患者眼内液检测的临床分析

目的 探讨眼内液检测在葡萄膜炎中的临床应用。方法 对2020年6月至2021年11月在中国科学技术大学附属第一医院(安徽省立医院)就诊并进行眼内液检测的葡萄膜炎35例(35只眼)进行回顾性分析，分析其临床特征，并结合眼内液检测做出最后诊断。结果 根据患者及临床诊断需要，35例葡萄膜炎中，29例进行病毒核酸检测，29例进行细胞因子检测，5例进行宏基因检测。29例进行病毒核酸检测样本中，17例样本检测结果阳性，以为水痘-带状疱疹病毒感染为主。29例进行细胞因子检测样本中，白介素6(IL-6)和白介素8(IL-8)水平高于正常值例数较多，分别为29(100%)例和26例(90%)。有25例报告中进行了白介素10(IL-10)的检测，其中有18例(72%)高于正常值；所有检查结果的IL-10/IL-6<1,不支持B细胞淋巴瘤的诊断。5例进行宏基因检测样本中，4例样本检测结果阳性。结论 眼内液检测对感染性葡萄膜炎的病原学诊断具有重要意义。     

感染性眼内炎282例临床分析

目的 分析感染性眼内炎的病因和病原学特征，提高感染性眼内炎的认知和临床诊治水平。方法 回顾分析282例住院治疗的感染性眼内炎患者的临床和实验室检测资料。患者中，男性206例，占73.05%；女性76例，占26.95%。年龄最小2岁，最大79岁，平均年龄（36.0±11.2）岁。除14例病情较重，无法保存眼球者以外，其余268例患者进行了玻璃体标本的病原学检测。临床和实验室检查数据采用Chi-square检验和Kappa一致性检验方法进行统计学分析，以P＜0.05为差异有统计学意义。结果 282例感染性眼内炎患者中，眼外伤所致者177例，占62.77%；感染性角膜炎所致者61例，占21.63%；眼部手术后所致者32例，占11.35%；内源性眼内炎12例，占4.25%。不同病因所致的感染性眼内炎患者中，2～13岁者占眼外伤所致者的19.21%；14～50岁者占感染性角膜炎所致者的55.74%；51～79岁者占眼部手术后所致者的60.61%。病原体检测显示，真菌和葡萄球菌属各占30.63%，为本组感染性眼内炎患者的主要病原体。其中，致病细菌对环丙沙星敏感者最多，占81.81％；庆大霉素次之，占77.92％。结论 感染性眼内炎的主要致病因素是眼外伤，真菌和葡萄球菌属是导致眼内感染的主要病原体。明确感染性眼内炎的不同致病因素、病原学特征有助于提供恰当的临床治疗。     

基于环介导等温扩增方法高效检测模拟眼内液中铜绿假单胞菌的研究△

目的 建立一个稳定、高效检测眼内液中铜绿假单胞菌的体系。方法 采用环介导等温扩增(LAMP)技术,由0.1~0.2 mL眼内平衡盐溶液(BSS)模拟眼内房水或玻璃体,利用铜绿假单胞菌gbca基因,特异性设计5组LAMP 引物,并对扩增方法进行关键试剂的优化,检测眼内液中铜绿假单胞菌。结果 该扩增方法经过优化后,最低可以检出100 fg/μL纯基因组DNA,接近荧光定量灵敏度,具有很好的重现性。此外该方法在检测眼内液中混合的菌落时呈现出良好的检测效果,可以检测出2.1×10² CFU/mL,极具应用价值。结论 建立了一个稳定、高效、灵敏的恒温扩增体系,可以应用于眼内液中铜绿假单胞菌的检测。     

彩色多普勒在眼科的应用进展

眼血流测定在眼部多种疾病的研究中得到越来越多的应用.彩色多普勒成像(CDI)技术可检测眼球后部血管的位置、血流方向和流速,为研究眼球的组织结构、眼球后血流动力学特征以及病理状态下的改变和临床疗效观察提供了一种简便、直观、无创、可重复、可连续动态观察的检查方法.对眼部血管性疾病和肿瘤的诊断与鉴别诊断有重要价值.现就CDI在青光眼、脉络膜和视网膜循环性疾病、眼内和眼眶占位性疾病中的应用进展综述如下.     

葡萄膜炎诊断中正确应用眼内液检测

眼内液检测是指采用房水或玻璃体样本进行的实验室检查，可用于某些类型葡萄膜炎的诊断。本文概述了眼内液检测的具体内容和检查结果的解读，阐述了其在葡萄膜炎诊断中的适应证，并对目前此类检查中存在的误区及其带来的后果进行了剖析。绝大多数葡萄膜炎患者可根据病史、临床检查及辅助检查即可做出诊断，并不需要进行眼内液检测，仅在少数特定的炎症如眼内炎、肿瘤所致的伪装综合征、眼弓形虫病、眼弓蛔虫病等才需进行眼内液检测。过度依赖眼内液检测查找病因不仅增加了感染的风险，对多数葡萄膜炎的病因诊断帮助不大甚至误诊误治。     

中药单体对眼部新生血管性疾病治疗作用的研究现状

概述中药单体在眼部新生血管性疾病中的应用现状。通过检索近十余年来国内外相关研究文献,进行归纳和总结,概述中药单体抑制眼内新生血管的研究进展,阐述中医药在眼内新生血管研究领域具有的潜在优势,为眼部新生血管性疾病的研究及治疗提供新的思路和参考。     

Polymerase chain reaction and Goldmann-Witmer coefficient analysis are complimentary for the diagnosis of infectious uveitis

PURPOSE: To determine the relative contribution of the analysis of intraocular antibody production and the polymerase chain reaction (PCR) in aqueous humor (AH) to the diagnosis of infectious uveitis. DESIGN: Retrospective case-control study. METHODS: Paired AH and serum samples from 230 patients suspected of infectious uveitis were examined for intraocular antibody production against herpes simplex virus (HSV), varicella zoster virus (VZV), and Toxoplasma gondii by calculating the Goldmann-Witmer coefficient (GWC). In addition, AH samples were investigated by real-time PCR to determine the presence of microbial DNA. RESULTS: Positive results were obtained in 54 cases (23%): 13 HSV (24%), 16 VZV (30%), and 25 T gondii (46%). Of these, 23 (43%) were positive for both GWC and PCR, 26 (48%) only for GWC, and 5 (9%) only for PCR. With PCR as the sole diagnostic approach, a correct diagnosis of the infectious etiology would have been missed in 34% of cases for the herpes viruses and in 64% for T gondii. Analysis of the relationship between a positive laboratory diagnosis and the time of sampling after onset of ocular disease demonstrated that intraocular antibody production was found throughout the course of the diseases. Viral DNA was more readily detected early in infection. In contrast, T gondii nucleic acid was not detected until 3 weeks after onset of ocular disease. CONCLUSIONS: Analysis of intraocular antibody production contributed considerably to the etiological diagnosis of infectious uveitis, most notably of ocular toxoplasmosis early after onset of disease. Therefore, both PCR and GWC determination might be performed for comprehensive diagnosis of intraocular infections.     

Novel infectious agents causing uveitis

In any patient with uveitis, an infectious cause should be ruled out first. The differential diagnosis includes multiple well-known diseases including herpes, syphilis, toxoplasmosis, tuberculosis, bartonellosis, Lyme disease, and others. However, clinician should be aware of emerging infectious agents as potential causes of systemic illness and also intraocular inflammation. Air travel, immigration, and globalization of business have overturned traditional pattern of geographic distribution of infectious diseases, and therefore one should work locally but think globally. This review recapitulates the systemic and ocular manifestations of several emergent infectious diseases relevant to the ophthalmologist including Rickettsioses, West Nile virus infection, Rift valley fever, Dengue fever, and Chikungunya. Retinitis, chorioretinitis, retinal vasculitis, and optic nerve involvement have been associated with these emergent infectious diseases. The diagnosis of any of these infections is usually based on pattern of uveitis, systemic symptoms and signs, and specific epidemiological data and confirmed by detection of specific antibody in serum. A systematic ocular examination, showing fairly typical fundus findings, may help establish an early clinical diagnosis, which allows prompt, appropriate management.     

山东省感染性眼内炎病原菌谱特点及药物敏感特征的临床研究

目的分析山东省感染性眼内炎的病原菌谱分布及药物敏感性特征。 方法回顾性分析2013年1月至2020年12月于山东第一医科大学附属眼科医院(山东省眼科医院)诊断为疑似感染性眼内炎350例(350只眼)患者的病例资料。其中,男性272例(272只眼),女性78例(78只眼);年龄1~87岁,平均年龄(44.4±20.0)岁。均为单眼发病。抽取所有患者的房水和(或)玻璃体(统称眼内液)进行涂片和体外培养实验。使用全自动微生物分析仪进行鉴定分离到的细菌和念珠菌,并行药物敏感性试验。采用WHONET5.6软件分析眼内炎患者的临床特点、病原菌分布以及主要病原菌的耐药性情况。感染性眼内炎病原菌分布和致病因素分布采用例数和百分比表示。 结果疑似感染性眼内炎350例患者中外源性眼内炎者和内源性眼内炎者分别为317例和33例,分别占90.6%和9.4%。房水和玻璃体均采集者269例,只采集房水者81例。涂片阳性者99例,阳性率为28.3%。涂片检出菌中革兰氏阳性球菌者、革兰氏阴性菌者、革兰氏阳性杆菌及真菌者分别为67例、8例、8例及16例,分别占67.7%、8.1%、8.1%及16.1%。病原微生物培养标本中病原菌阳性者135例,阳性率为38.6%,共分离得到病原菌144株。其中,革兰氏阳性球菌者、革兰氏阳性杆菌者、革兰氏阴性杆菌者及真菌者分别为87例、12例、15例及30例,分别占60.4%、8.3%、10.5%及20.8%。以眼内液涂片和(或)病原菌培养阳性为感染性眼内炎诊断的金标准,在350例全部疑似患者中,有211例患者眼内液涂片和(或)病原菌培养阳性,最终确诊为感染性眼内炎,感染阳性率为60.3%。病原菌中以凝固酶阴性的葡萄球菌居首位,其次为链球菌。真菌中主要以镰刀菌和曲霉菌为主。分离菌株对左氧氟沙星的敏感性最高达39.3%~84.6%。 结论2013年至2020年间,眼外伤是山东地区感染性眼内炎的主要致病因素,革兰氏阳性球菌尤其凝固酶阴性的葡萄球菌是感染性眼内炎的主要病原菌。     

Algorithmic approach in the diagnosis of uveitis

Uveitis is caused by disorders of diverse etiologies including wide spectrum of infectious and non-infectious causes. Often clinical signs are less specific and shared by different diseases. On several occasions, uveitis represents diseases that are developing elsewhere in the body and ocular signs may be the first evidence of such systemic diseases. Uveitis specialists need to have a thorough knowledge of all entities and their work up has to be systematic and complete including systemic and ocular examinations. Creating an algorithmic approach on critical steps to be taken would help the ophthalmologist in arriving at the etiological diagnosis.     

A new era of uveitis: impact of polymerase chain reaction in intraocular inflammatory diseases

Uveitis is a sight-threatening intraocular inflammatory disorder which may occur from both infectious and non-infectious or autoimmune causes. The frequency of infectious uveitis and autoimmune uveitis varies depending on countries and regions. According to a nationwide survey conducted by the Japanese Ocular Inflammation Society, infectious and non-infectious uveitis accounted for 16.4 and 50.1% of new patients, respectively while the remaining 33.5% of new uveitis cases were not classified or were idiopathic uveitis. Infectious uveitis is particularly important because it causes tissue damage to the eye and may result in blindness unless treated. However, it can be treated if the pathogenic microorganisms are identified promptly and accurately. Remarkable advancements in molecular and immunological technologies have been made in the last decade, and the diagnosis of infectious uveitis has been greatly improved by the application of molecular and immunological investigations, particularly polymerase chain reaction (PCR). PCR performed on a small amount of ocular samples provides a prompt, sensitive, and specific molecular diagnosis of pathogenic microorganisms in the eye. This technology has opened a new era in the diagnosis and treatment of uveitis, enabling physicians to establish new clinical entities of uveitis caused by infectious microorganisms, identify pathogens in the eyes of many patients with uveitis, and determine prompt diagnosis and appropriate therapy. Here we review the PCR process, new PCR tests specialized for ocular diseases, microorganisms detected by the PCR tests, diseases in the eye caused by these microorganisms, and the clinical characteristics, diagnosis, and therapy of uveitis.     

Intraocular anti-Toxoplasma gondii IgA antibody production in patients with ocular toxoplasmosis

PURPOSE: To investigate the immunoglobulin classes associated with the intraocular anti-Toxoplasma gondii antibody response during clinical ocular toxoplasmosis and to determine which immunoglobulin class is most helpful in the diagnosis of this disease. METHODS: Paired serum and intraocular fluid samples from 155 patients who had uveitis were tested for intraocular anti-T. gondii IgG, IgA, and IgM antibody production. The presence of T. gondii DNA was determined by polymerase chain reaction. Patients were divided into two groups, based on the initial clinical diagnosis; group 1 included 78 patients with presumed ocular toxoplasmosis, and group 2 included 77 patients with uveitis that was not clinically suspected to be ocular toxoplasmosis. Samples from 27 nonuveitis patients who underwent intraocular surgery were used as control subjects. The final diagnosis was based on the clinical course and interpretation of laboratory tests. RESULTS: A final diagnosis of ocular toxoplasmosis was made in 88 of 155 patients (group 1, 68; group 2, 20). Among these patients, 65% had intraocular IgG production, 52% had intraocular IgA production, 37.5% had both IgG and IgA production, 27% had IgG production only, and 15% had IgA production only. Of the 13 patients tested, only one had intraocular IgM production. Intraocular IgA could not be detected in patients who had final diagnoses other than ocular toxoplasmosis or in control subjects. A positive polymerase chain reaction combined with a test that was positive for intraocular IgG production had a sensitivity of 77%, which increased to 91% after the detection of intraocular IgA production was added. CONCLUSIONS: Immunoglobulin G is the major class involved in the humoral immune response against the T. gondii parasite, followed by IgA. The determination of IgA production is useful as an additional test in the diagnosis of ocular toxoplasmosis.     

眼外伤后眼内组织骨化

目的 观察眼外伤后眼内组织骨化症的临床和病理特征。方法 对2例眼外伤后眼内组织骨化症患者做眼科检查，并行手术治疗，将摘除组织作病理检查。结合文献对本病的病因、发生、临床特点、诊断及鉴别诊断进行分析讨论。结果 2例均有10年以上的穿孔性外伤史。患眼视功能逐渐减退，最后为无光感及眼球萎缩。2例均进行眼内容摘除术，经病理检查证实为眼内组织骨化。结论 眼内组织骨化症是严重眼外伤的并发症。     

Prenatal genetic diagnosis of retinoblastoma – clinical correlates on follow-up

Retinoblastoma is the most common malignant intraocular tumor in pediatric age group if undetected leads to ocular mortality. Prenatal diagnosis is an emerging technology to detect fatal diseases in utero such that subsequent management is planned to reduce the ocular morbidity. We describe a case demonstrating the importance of prenatal diagnosis in a child with a strong family history of retinoblastoma and importance of a long-term clinical follow-up in these cases.