糖尿病患者真菌性角膜炎5例临床与真菌学分析

真菌性角膜炎是眼科致盲率较高的眼病之一,近年来发病有逐渐上升趋势。糖尿病患者合并真菌性角膜炎少有报道。本文报道糖尿病患者并发真菌性角膜炎5例,分析其临床和真菌学特点。     

应用ROC曲线分析泪液(1-3)-β-D葡聚糖对真菌性角膜炎的诊断价值

目的探讨真菌性角膜炎患者泪液(1-3)-β-D葡聚糖的阈值,通过受试者工作特征曲线(ROC)评价其对真菌性角膜炎的诊断价值。方法分别测定100例真菌性角膜炎患者和100例健康志愿者泪液中(1-3)-β-D葡聚糖水平,应用ROC曲线评估泪液(1-3)-β-D葡聚糖诊断真菌性角膜炎的最佳临界值及其灵敏度、特异性。结果泪液(1-3)-β-D葡聚糖诊断真菌性角膜炎的ROC曲线下面积为0.992,阈值为23.82pg/mL,灵敏度为97.0%,特异性为98.0%。结论泪液(1-3)-β-D葡聚糖对真菌性角膜炎的诊断具有较高的灵敏度和特异性,具有一定临床应用价值。     

真菌性角膜炎的实验室诊断

真菌性角膜炎是一种危害较大的角膜感染性疾病,近十年发病率不断上升,早期、及时诊断对于提高该病的治愈率,降低致残率非常重要。典型的角膜溃疡、明确的角膜外伤史可以帮助诊断真菌性角膜炎。实验室诊断非常重要,找到致病真菌是确诊的依据。本文主要介绍真菌性角膜炎的各种实验室诊断方法的优缺点以及近年的进展。     

角膜激光共聚焦显微镜在单眼感染性角膜炎疾病诊断中的应用

摘要 目的：探讨角膜激光共聚焦显微镜在单眼感染性角膜炎疾病诊断中的应用价值。方法：回顾性研究2020年6月到2021年6月选择在本院诊治的单眼感染性角膜炎疾病患者62例,所有患者都给予角膜激光共聚焦显微镜检查,记录影像学特征并判断诊断价值（以病原学诊断为金标准）。结果：真菌性角膜炎在角膜激光共焦显微镜下的病变区纵横交错的高反射的真菌菌丝或高反光细长颗粒状的孢子,细菌性角膜炎的病变处会聚集活化的树突状细胞及大量的炎症细胞,病毒性角膜炎的基底膜下神经纤维密度、神经主干的分支数减少,棘阿米巴性角膜炎的包囊表现为圆形高反光厚壁结构。角膜激光共聚焦显微镜判断为病毒性角膜炎17例,诊断病毒性角膜炎的敏感性与特异性为94.4 %和100.0 %;角膜激光共聚焦显微镜判断为棘阿米巴性角膜炎4例,诊断棘阿米巴性角膜炎的敏感性与特异性为94.4 %和100.0 %;角膜激光共聚焦显微镜判断为细菌性角膜炎21例,诊断细菌性角膜炎的敏感性与特异性为95.5 %和97.5 %;角膜激光共聚焦显微镜判断为真菌性角膜炎20例,诊断真菌性角膜炎的敏感性与特异性为94.4 %和93.2 %。ROC曲线分析显示角膜激光共聚焦显微镜诊断细菌性角膜炎、真菌性角膜炎、病毒性角膜炎、棘阿米巴性角膜炎的曲线下面积分别为0.525、0.579、0.777、0.731。结论：角膜激光共聚焦显微镜在单眼感染性角膜炎疾病诊断中的应用能较好的区分细菌性角膜炎、真菌性角膜炎、病毒性角膜炎、棘阿米巴性角膜炎,具有良好的诊断敏感性与特异性。     

糖尿病真菌性角膜炎患者临床特征及病原学分析CSCD

目的调查与探讨糖尿病真菌性角膜炎患者临床特征及病原学状况,发现影响糖尿病真菌性角膜炎患者预后的主要危险因素。方法采用回顾性研究方法,选择2013年2月~2016年5月在我院眼科确诊的,具有完整病例资料和随访资料的真菌性角膜炎患者120例作为研究对象,包括合并糖尿病的真菌性角膜炎60例(糖尿病组)和无糖尿病的真菌性角膜炎60例(非糖尿病组),调查所有患者的人口学特征、临床症状、病原学检查结果,同时记录患者的预后情况并进行危险因素分析。结果两组患者的性别、体重指数、病程等对比,差异无统计学意义(P>0.05),糖尿病组糖尿病病史平均为(8.14±2.11)a,糖尿病组的年龄和角膜感染程度评分与非糖尿病组相比,差异有统计学意义(P<0.05)。在糖尿病组中,镰刀菌属10例(16.7%),曲霉菌属40例(66.7%),念珠菌属10例(16.7%);在非糖尿病组中,镰刀菌属25例(41.7%),曲霉菌属30例(50.0%),念珠菌属5例(8.3%),两两比较差异都有统计学意义(P<0.05)。糖尿病组与非糖尿病组的治疗总有效率分别为86.7%和98.3%,糖尿病组的治疗总有效率明显差于非糖尿病组(P<0.05)。多分变量Logistic回归显示浸润病灶、前房积脓、曲霉菌为影响糖尿病真菌性角膜炎预后的主要危险因素(P<0.05)。结论糖尿病真菌性角膜炎患者相对于非糖尿病患者而言主要表现为发病年龄偏大、角膜感染较重、预后差的特点,曲霉属为其主要病原菌。浸润病灶、前房积脓和曲霉菌为影响糖尿病真菌性角膜炎预后的主要危险因素。     

束状刺盘孢角膜炎的实验研究

目的初步研究束状刺盘孢感染兔角膜的临床表现和组织病理学特征。方法选择15只新西兰白兔,应用基质内直接注射法建立模型。肉眼观察角膜变化,并在接种后第5 d、14 d、21 d处死家兔,对角膜行HE染色和PAS染色,观察其组织病理学改变。结果兔束状刺盘孢角膜炎动物模型成功建立,肉眼可以见到典型的真菌性角膜炎的表现。组织病理学检查,在感染早期即可看到大量真菌病原体生长,菌丝垂直于角膜基质,后期出现新生血管和瘢痕。结论角膜基质内直接注射法复制真菌性角膜炎动物模型成功率高、稳定。兔束状刺盘孢角膜炎有典型的真菌性角膜炎的表现,组织学中菌丝垂直于角膜基质生长。     

糖尿病真菌性角膜炎患者临床特征及病原学分析

目的调查与探讨糖尿病真菌性角膜炎患者临床特征及病原学状况,发现影响糖尿病真菌性角膜炎患者预后的主要危险因素。方法采用回顾性研究方法,选择2013年2月~2016年5月在我院眼科确诊的,具有完整病例资料和随访资料的真菌性角膜炎患者120例作为研究对象,包括合并糖尿病的真菌性角膜炎60例(糖尿病组)和无糖尿病的真菌性角膜炎60例(非糖尿病组),调查所有患者的人口学特征、临床症状、病原学检查结果,同时记录患者的预后情况并进行危险因素分析。结果两组患者的性别、体重指数、病程等对比,差异无统计学意义(P0.05),糖尿病组糖尿病病史平均为(8.14±2.11)a,糖尿病组的年龄和角膜感染程度评分与非糖尿病组相比,差异有统计学意义(P0.05)。在糖尿病组中,镰刀菌属10例(16.7%),曲霉菌属40例(66.7%),念珠菌属10例(16.7%);在非糖尿病组中,镰刀菌属25例(41.7%),曲霉菌属30例(50.0%),念珠菌属5例(8.3%),两两比较差异都有统计学意义(P0.05)。糖尿病组与非糖尿病组的治疗总有效率分别为86.7%和98.3%,糖尿病组的治疗总有效率明显差于非糖尿病组(P0.05)。多分变量Logistic回归显示浸润病灶、前房积脓、曲霉菌为影响糖尿病真菌性角膜炎预后的主要危险因素(P0.05)。结论糖尿病真菌性角膜炎患者相对于非糖尿病患者而言主要表现为发病年龄偏大、角膜感染较重、预后差的特点,曲霉属为其主要病原菌。浸润病灶、前房积脓和曲霉菌为影响糖尿病真菌性角膜炎预后的主要危险因素。     

茄病镰刀菌感染致角膜溃疡1例

报告1例由茄病镰刀菌感染引起的角膜溃疡。患者男性,40岁,木工,左眼红、痛、流泪、视力下降10 d。经过角膜清创,系统性及局部抗真菌治疗后好转。致病菌株经真菌学鉴定为茄病镰刀菌,它是真菌性角膜炎的常见致病菌之一。真菌性角膜炎病程进展迅速,能否及早诊治关系预后。真菌学检查是确诊的依据,临床应予重视。     

糖尿病患者感染轮枝镰刀菌致角膜炎1例

报道糖尿病患者感染轮枝镰刀菌致真菌性角膜炎1例。患者男性,48岁,因右眼红,痛,视力下降20天就诊。患者有2型糖尿病史近10年,注射胰岛素血糖一直控制不佳。刮取角膜分泌物标本进行真菌荧光染色镜检和培养。对培养的菌株进行形态学(乳酸酚棉兰染色)和分子生物学鉴定。真菌荧光染色镜检阳性,镜下可见分支分隔的菌丝及鹿角状菌丝。真菌培养阳性,乳酸酚棉兰染色镜下可见多数棒状的小分生孢子。ITS和Beta-tubulin基因序列分析鉴定为轮枝镰刀菌。依据临床和实验室检查确定诊断右眼角膜炎(真菌性),2型糖尿病。给予患者那他霉素等滴眼液点右眼局部抗炎抗真菌对症处理,用药治疗后再次行眼科检查,患者右眼症状较治疗前明显好转。出院后继续抗真菌抗炎治疗2周,门诊随访。     

张家口地区真菌性角膜炎病原菌的临床分析

目的：分析张家口地区真菌性角膜炎的病原学及流行病学特征,为其临床诊治提供一定的参考依据。方法：回顾性分析2010年12月-2012年6月我院收治的97例真菌性角膜炎患者的临床资料,分析其病原菌的构成、诊断方法及药物敏感性情况。结果：总共鉴定出84株真菌,分属12个属、23个种,其中以镰刀菌属感染最为多见（51．2％）,其次为曲霉属（14．3％）和假丝酵母属（10．7％）;感染者以男性劳动者居多,外伤是最常见的诱因,秋冬季为高发季节;不同菌种对药物的敏感性存在区别,主要致病菌种对两性霉素B和纳他霉素的敏感性均达70％以上,较其它抗真菌药物高。结论：镰刀菌为本地区真菌性角膜炎的主要致病菌,病原体种类和构成与其他北方地区无明显区别。两性霉素B和纳他霉素为大多数菌种的敏感药物,可作为本地区临床预防和治疗的首选药物。     

Characterization of Edenia gomezpompae Isolated from a Patient with Keratitis

Edenia gomezpompae, isolated from a patient suffering from keratitis, who was an agricultural worker with a 4-year history of diabetes mellitus, is described. Fungal hyphae were detected in corneal scrapings after microscopic examination and repeated culture. Isolates were identified by morphology and by sequencing the internal transcribed spacer region of ribosomal DNA. Chlamydospore characteristics of E. gomezpompae are described for the first time. The in vitro antifungal susceptibilities of the isolate were tested using five antifungal agents. With the exception of fluconazole, the other drugs (natamycin, amphotericin B, itraconazole, and voriconazole) assayed were highly effective against this fungus. The keratitis was cured after debridement of the corneal ulcer and treatment with 5 % natamycin. After 3 months, when the patient was reexamined, the corneal ulcer showed signs of healing, with no recurrence. To our knowledge, this is the first reported case of keratitis caused by E. gomezpompae, as well as being the first known case of disease in humans caused by this species.     

Rare Fungal Keratitis Caused by Coprinellus Radians

Mycopathologia - A case of fungal keratitis due to Coprinellus radians is reported. To our knowledge, fungal keratitis caused by this species was rare. Fungal hyphae were detected in corneal...     

Galectin-1 reduces the severity of herpes simplex virus-induced ocular immunopathological lesions

Stromal keratitis is a chronic immunopathological lesion of the eye caused by HSV-1 infection that can result in blindness. Because the inflammatory lesions are primarily orchestrated by Th1 cells, and to a lesser extent by Th17 cells, inhibiting their activity represents a useful form of therapy. In this study we evaluated the therapeutic potential of galectin-1 (gal-1), an endogenous lectin that in some autoimmune diseases was shown to suppress the functions of Th1 and Th17 cells. Treatment was begun at different times after ocular infection with HSV and the outcome was assessed clinically as well as for effects on various immune parameters. Treatment with recombinant gal-1 significantly diminished stromal keratitis lesion severity and the extent of corneal neovascularization. Treated mice had reduced numbers of IFN-γ- and IL-17-producing CD4(+) T cells, as well as neutrophil infiltration in the cornea. Furthermore, disease severity was greater in gal-1 knockout mice compared with their wild-type counterparts. The many effects of gal-1 treatment include reduction in the production of proinflammatory cytokines and chemokines, increased production of IL-10, and inhibitory effects on molecules involved in neovascularization. To our knowledge, our findings are the first to show that gal-1 treatment represents a useful approach to control lesion severity in a virally induced immunopathological disease.     

Acanthamoeba: biology and increasing importance in human health

Acanthamoeba is an opportunistic protozoan that is widely distributed in the environment and is well recognized to produce serious human infections, including a blinding keratitis and a fatal encephalitis. This review presents our current understanding of the burden of Acanthamoeba infections on human health, their pathogenesis and pathophysiology, and molecular mechanisms associated with the disease, as well as virulence traits of Acanthamoeba that may be targets for therapeutic interventions and/or the development of preventative measures.     

The increasing importance of Acanthamoeba infections

Free-living amebae belonging to the genus Acanthamoeba are the causative agents of granulomatous amebic encephalitis, a chronic progressive disease of the central nervous system, and of amebic keratitis, a chronic eye infection. Granulomatous amebic encephalitis occurs more frequently in immunocompromised patients while keratitis occurs in healthy individuals. The recent increased incidence in Acanthamoeba infections is due in part to infection in patients with acquired immune deficiency syndrome, while that for keratitis is due to the increased use of contact lenses. Understanding the mechanism of host resistance to Acanthamoeba is essential since the amebae are resistant to many therapeutic agents. Studies in our laboratory as well as from others have demonstrated that macrophages from immunocompetent animals are important effector cells against Acanthamoeba. We have demonstrated also that microglial cells, resident macrophages of the brain, elicit cytokines in response to A. castellanii. Neonatal rat cortical microglia from Sprague-Dawley rats co-cultured with A. castellanii produced mRNA for the inflammatory cytokines, interleukin 1alpha, interleukin 1beta, and tumor necrosis factor alpha. In addition, scanning and transmission electron microscopy revealed that microglia ingested and destroyed A. castellanii in vitro. These results implicate macrophages as playing an effector role against Acanthamoeba and suggest immune modulation as a potential alternative therapeutic mode of treatment for these infections.     

IL-17 produced by Th17 cells alleviates the severity of fungal keratitis by suppressing CX43 expression in corneal peripheral vascular endothelial cells

Fungal keratitis is a relatively common ocular disease requiring positive medical management combined with surgical intervention. Interleukin-17 (IL-17) was reported to promote the activation and mobilization of neutrophile granulocyte to foci of inflammation. This study investigated the effect of IL-17 production from Th17 cells on the progression of fungal keratitis. A mouse model of fungal keratitis induced by Candida albicans was successfully constructed to detect infiltration of inflammatory cells in corneal tissues by hematoxylin-eosin (HE) staining and immunohistochemistry. Fungal load capacity of mouse cornea was also detected. The regulatory role of IL-17 in fungal keratitis with the involvement of CX43 was investigated with the relevant expression of inflammatory factors detected and activation of vascular endothelial cells assessed. Furthermore, in vivo experiment was also performed to confirm the role of CX43 in keratitis. Mice with fungal keratitis showed increased level of inflammatory cytokines and infiltration of inflammatory cells. Silencing IL-17 in Th17 cells and overexpressing CX43 could inhibit the activation of vascular endothelial cells. Besides, CX43 knockdown in vivo alleviated fungal keratitis in mice. The possible mechanism of the above findings could be IL-17 inhibiting the level of CX43 through the AKT signaling pathway. Taken together, IL-17 could inhibit the occurrence and development of fungal keratitis by suppressing CX43 expression through the AKT signaling pathway. Therefore, this study provides a potential target for the treatment of fungal keratitis.     

Staphylococcus aureus Keratitis: A Review of Hospital Cases

Background

Methicillin-resistant Staphylococcus aureus (MRSA) infection is an important public health issue. The study aimed to characterize the patient demographics, clinical features, antibiotic susceptibility, and clinical outcomes of keratitis caused by S. aureus, and to make a comparison between MRSA and methicillin-sensitive S. aureus (MSSA) isolates.

Methodology/Principal findings

Patients (n = 59) with culture-proven S. aureus keratitis treated in Chang Gung Memorial Hospital between January 1, 2006, and December 31, 2010, were included in our study. Patients'' demographic and clinical data were retrospectively reviewed. Twenty-six MRSA (44%) and 33 MSSA (56%) isolates were collected. The MRSA keratitis was significantly more common among the patients with healthcare exposure (P = 0.038), but 46.2% (12/26) of patients with MRSA keratitis were considered to have community-associated infections. All isolates were susceptible to vancomycin. MRSA isolates were significantly more resistant to clindamycin, erythromycin, and sulfamethoxazole/trimethoprim. Ocular surface disease was a significant risk factor for MRSA keratitis (P = 0.011). Visual outcome did not differ significantly between the MRSA and MSSA groups. However, age (B = 0.01, P = 0.035, 95% confidence interval [CI]: 0.001–0.019) and visual acuity at presentation (B = 0.749, P<0.001, 95% CI: 0.573–0.926) were significantly correlated with visual outcome.

Conclusions/Significance

Ocular surface disease is an important predisposing factor for S. aureus keratitis, especially for MRSA infections. Advanced age and poor visual acuity at presentation are important prognostic indicators for poor visual outcome in S. aureus keratitis. Oxacillin resistance may not be a significant prognostic indicator.     

Enzymes of cyclic nucleotide degradation in eye tissues during experimental ocular herpes

Activity of phosphodiesterases disintegrating cAMP and cGMP in the cornea, sclera and ciliary body was investigated in health and in different stages of experimental herpetic keratitis. The problems concerning the role of the cyclase system in the pathogenesis of herpetic keratitis and the possibility of applying some of the drugs to the disease treatment are discussed.     

Immunological inter-strain crossreactivity correlated to 18S rDNA sequence types in Acanthamoeba spp

Various species of the genus Acanthamoeba have been described as potential pathogens; however, differentiation of acanthamoebae remains problematic. The genus has been divided into 12 18S rDNA sequence types, most keratitis causing strains exhibiting sequence type T4. We recently isolated a keratitis causing Acanthamoeba strain showing sequence type T6, but being morphologically identical to a T4 strain. The aim of our study was to find out, whether the 18S rDNA sequence based identification correlates to immunological differentiation. The protein and antigen profiles of the T6 isolate and three reference Acanthamoeba strains were investigated using two sera from Acanthamoeba keratitis patients and one serum from an asymptomatic individual. It was shown, that the T6 strain produces a distinctly different immunological pattern, while patterns within T4 were identical. Affinity purified antibodies were used to further explore immunological cross-reactivity between sequence types. Altogether, the results of our study support the Acanthamoeba 18S rDNA sequence type classification in the investigated strains.     

Genotyping of pathogenic Acanthamoebae isolated from clinical samples in Greece--report of a clinical isolate presenting T5 genotype

Amoebae belonging to the genus Acanthamoeba are potentially pathogenic to humans, causing mainly amoebic keratitis. Pathogenic ability of the 15 known Acanthamoeba genotypes is under investigation. We report that four out of five cases of amoebic keratitis studied in Greece, present T4 sequence type, while the remaining one presents T5 sequence type (Acanthamoeba lenticulata), which is the second most frequent genotype found among environmental samples. Thus, it is confirmed, for the first time to our knowledge, that A. lenticulata can cause keratitis. However the reason that it is under represented in clinical samples compared to environmental ones is unknown.