真菌性角膜炎的共焦显微镜影像学特点分析

目的探讨应用共焦显微镜观察真菌性角膜炎患者角膜各层的活体形态学特点。方法对65例（65只眼）真菌性角膜炎患者的病灶进行活体共焦显微镜检查,应用NAVIS软件测量、分析角膜各层菌丝、孢子形态、直径、密度。结果通过连续共焦扫描及焦点分析：65例中共焦显微镜显示：①10例显示上皮层下高反射直径2μm～4μm的树枝状菌丝;②27例显示浅、中基质层杂乱分布的直、长线状菌丝,直径约3μm～6μm,长度150μm～300μm。③25例显示浅、中基质层弥漫分布的高反射短、段状菌丝,直径3μm～6μm,长度40μm～60μm。④3例显示：直径12μm～15μm的高亮度圆形、椭圆形实心球体孢子。菌丝侵入分布密度与炎性细胞分布密度成负相关（r=-0.019;p=0.026）。结论共焦显微镜下真菌性角膜炎菌丝有不同的影像学特点,随着菌丝侵入角膜深度不同,菌丝形态亦不相同。明确角膜基质深层菌丝形态在共焦显微镜检查中尤显重要。     

激光共焦显微镜在棘阿米巴角膜炎诊断及治疗中的初步临床研究

目的 应用激光共焦显微镜对棘阿米巴性角膜炎镜下特征进行研究,探讨其在该病诊断和治疗中的应用价值.方法 应用激光共焦显微镜对拟诊为棘阿米巴性角膜炎患者9例(9只眼)进行观察,分析包囊的形态特点和分布特征;观察药物治疗过程中包囊、炎性细胞及树突细胞的变化情况;同时对每例行角膜刮片和培养,3例术眼行病理检查.结果 9例激光共焦显微镜下包囊多于浅中基质层,圆形双壁,边境清晰.7例多见包囊的串珠样分布.角膜环形浸润患者,环形浸润处炎性细胞、树突细胞密度高于中央,包囊分布无明显差异.角膜中央基质浸润患者,病灶中央与周边包囊、炎性细胞及树突细胞密度无明显差异.角膜中央基质浸润患者镜下病灶区早期以树突细胞及炎性细胞为主.抗棘阿米巴药物治疗1个月后,树突细胞略有下降,炎性细胞及包囊密度下降显著,包囊散在分布.2个月后查见少量炎性细胞及树突细胞,包囊极少查见.4例角膜刮片阳性,6例培养阳性.3例病理阳性.结论 激光共焦显微镜下棘阿米巴包囊多呈串珠样分布.观察药物治疗过程中包囊、炎性细胞和树突细胞的变化情况,可指导临床用药.     

应用激光共焦显微镜诊断真菌性角膜炎的临床研究

目的:应用激光共焦显微镜观察真菌性角膜炎患者图像特点及菌丝和孢子检出率,探讨激光共焦显微镜检查在真菌性角膜炎临床诊断中的意义。方法:对41例41眼经门诊确诊为真菌性角膜炎的患者行角膜激光共焦显微镜检查,观察不同治疗期真菌性角膜炎患者活体角膜各层图像特点。结果:共焦显微镜下真菌性角膜炎患者图像有如下特点:(1)病变部位角膜各层形态结构破坏明显;角膜上皮至基质不同程度水肿;炎细胞浸润;神经结构破坏;基质层结构紊乱,透过度降低;(2)真菌菌丝是本病的特异性诊断依据,不同菌种感染在镜下菌丝有不同的影像学特点;(3)不同病变时期和治疗阶段,图像有很大差异,真菌菌丝的检出并不是诊断本病的唯一依据。结论:激光共焦显微镜检查具有无创、及时等优点,在临床诊断真菌性角膜炎中有重要参考意义,尤其是指导临床早期诊断、合理治疗及评价预后。     

共焦显微镜对棘阿米巴角膜炎药物治疗后转归的临床观察

目的借助共焦显微镜来帮助判断棘阿米巴角膜炎患者在应用抗棘阿米巴药物治疗期间的效果及临床转归,为指导临床用药及选择治疗方案提供客观依据。方法对我院2005.2-2006.1住院拟诊为棘阿米巴角膜炎的5例5眼患者,给予抗棘阿米巴角膜炎药物治疗,在接受治疗的1w、2w、4w、8w分别对患者角膜病灶的特定位点进行共焦显微镜检查,从上皮下区或其浅基质层有活的滋养体和12um双壁的包囊及炎性细胞密度和形态的变化及空腔的出现来判断抗棘阿米巴药物治疗的用量频率,对病程较长、反复发作有疤痕者,共焦显微镜下未发现包囊及滋养体者,根据他的临床表现及病史作诊断性治疗,局部用药4m-6m。结果5例患者中4例治愈,视力不同程度提高,在治疗2m后复查共焦显微镜,未见明显包囊及滋养体及炎性细胞,可见疤痕样改变,偶见圆点状亮点,1例治疗1w后病情恶化,合并前部巩膜炎转杭州邵逸夫医院。结论共焦显微镜是目前对早期棘阿米巴角膜炎患者经药物治疗后疗效判断的一种无创伤性手段,其可作为指导临床抗棘阿米巴角膜炎药物治疗和判断预后的一种客观检查工具。     

共焦激光角膜显微镜在真菌性角膜炎诊断中的应用

目的评价共焦激光角膜显微镜在临床诊断真菌性角膜炎中的应用价值。方法用共焦激光角膜显微镜对临床拟诊为真菌性角膜炎的26例患者进行检查,同时行涂片检查真菌菌丝,对两组结果进行比较。结果26例患者中,根据病史、临床表现24例确诊为真菌性角膜炎,其中共焦激光角膜显微镜的确诊率为95.83%(23/24),角膜刮片的确诊率为83.33%(20/24)。结论共焦激光角膜显微镜是一种快速、有效、无损伤的活体检查方法,在真菌性角膜炎的早期诊断、治疗和研究中将起重要作用。     

真菌性角膜炎病原学诊断

目的分析角膜涂片细胞学检查、真菌培养、角膜共焦显微镜在真菌性角膜炎诊断中的应用价值。方法对324临床诊为真菌性角膜炎患者进行涂片细胞学检查、真菌培养和角膜共焦显微镜检查,并对其检查的阳性率进行汇分析,以探讨病原学诊断在真菌性角膜炎诊断中的应用价值。结果324份标本中,细胞学检查阳性者168例(51.85%),真菌培养阳性者212例(65.43%),角膜共焦显微镜检查阳性者277例(85.63%)。结论角膜共焦显微镜检查在真菌性角膜炎诊断中有重要意义,是一种快速、有效、无损伤、可以直接在活体角膜上观察真菌菌丝的检查方法,较真菌培养和细胞学检查对真菌性角膜炎的早期诊断,治疗具有更重要的临床价值。     

43例真菌性角膜炎的诊断及药物治疗临床分析

的分析经药物治疗的真菌性角膜炎病例的早期诊断方法和治疗过程,探讨药物治疗的可行性。方法收集2004年1月至2005年12月在我院经药物治疗的真菌性角膜炎43例,采用角膜刮片镜检加真菌培养鉴定进行诊断,联合2-3种抗真菌药物治疗、根据病情调整用药并观察治疗过程。结果43例经药物治疗的真菌性角膜炎中,35例镜检发现菌丝,39例真菌培养阳性;抗真菌药物治疗疗程18d-56d,5d-7d开始显效,表现为溃疡开始愈合、上皮逐渐修复。全部病例治愈后无复发。结论角膜刮片镜检是早期诊断真菌性角膜炎的有效方法;对发病早期、病情较轻的患者联合抗真菌药物治疗可取得良好疗效。     

共焦显微镜在临床诊断真菌性角膜炎中的应用

目的 评价共焦显微镜在临床诊断真菌性角膜炎中的应用价值。方法 用共焦显微镜对52例临床拟诊为真菌性角膜炎的患者进行检查，同时行角膜病灶刮片培养，对两组结果进行比较。结果 综合病史、临床表现及治疗结果，有45例临床诊断为真菌性角膜炎，其中32例角膜刮片培养到真菌，阳性率为71．11％(32／45)；43例共焦显微镜检查观察到真菌菌丝和／或孢子，阳性率为95．56％(43／45)。结论 共焦显微镜可以在活体角膜上直接观察到真菌菌丝，是一种快速、有效和无损伤的检查方法，对真菌性角膜炎的早期诊断、治疗和研究等具有重要的临床价值。     

镰孢菌属真菌性角膜炎的临床及共焦显微镜特征分析

目的 回顾性分析镰孢菌感染所致的真菌性角膜炎共焦显微镜影像特征及疗效.方法 将43例(43眼)临床确诊的镰孢菌感染所致的真菌性角膜炎进行裂隙灯显微镜及共焦显微镜检查,并详细记录.结果 43例镰孢菌感染所致的真菌性角膜炎裂隙灯显微镜观察:23例(92%)可见菌丝苔被,20例(80%)可见羽毛状浸润边缘,8例(18.6%)观察到前房积脓;共焦显微镜下显示:19例(44.2%)上皮层及浅基质层检查到长度50～200 μm,直径2～5 μm的树枝状菌丝;16例(37.2%)显示浅中基质层的杂乱分布的直、长线状菌丝,直径3～7 μm,长度150～300 μm.结论 菌丝苔被、羽毛状浸润边缘是镰孢菌属感染所致的真菌性角膜炎的主要临床特点.不同菌种镰孢菌属感染所致的真菌性角膜炎的共焦显微镜影像表现较为一致,可见典型的菌丝形态.     

激光共焦显微镜在真菌性角膜炎诊断中的应用

目的:探讨激光共焦显微镜HRT3-RCM在真菌性角膜炎诊断中的应用价值。方法:对临床拟诊真菌性角膜炎患者32例32眼行HRT3-RCM检查,同时进行角膜刮片细胞学检查。结果:在32例患者中,24例在角膜组织刮片中查到了菌丝,阳性率为75%;30例在激光共焦显微镜下观察到真菌菌丝,阳性率为94%。结论:HRT3-RCM是利用激光作为光源的新一代共焦显微镜,是一种快速、有效、无创的检查方法,可用于真菌性角膜炎的早期病原学诊断以及治疗观察。     

Antifungal chemotherapy for fungal keratitis guided by in vivo confocal microscopy

Background To evaluate antifungal chemotherapy in patients with fungal keratitis guided by in vivo confocal microscopy. Methods A total of 121 patients (121 eyes) with fungal keratitis were enrolled in this study. Confocal microscopy was performed in real time after topical and/or oral antifungal chemotherapy. Hyphal density and morphology, composition of inflammatory cells, and appearance of corneal stromal cells at the central and peripheral corneal lesions were recorded. Antifungal therapy discontinued at 1 week after hyphae and inflammatory cells could not be detected, and affected corneal stromal cells became visible. Results Successful outcomes were achieved in 110 patients (90.9%). By confocal microscopy, we observed the gradual decrease of hyphae-positive sites and hyphal density during the chemotherapy. The inflammatory cells reduced in number and heterogeneity, while corneal stromal cells recovered. The antifungal drugs were tapered according to the changes in hyphae, inflammatory cells, and corneal stromal cells. There was no fungal recurrence during the 2-month follow-up period. The other 11 patients (9.1%) had deteriorated infection within 1 week of antifungal therapy, and therefore were subjected to corneal transplantation. Conclusions In vivo confocal microscopy appears to be an effective approach to guide antifungal chemotherapy. It allows comprehensive evaluation of hyphae, inflammatory cells, and corneal stromal cells in real time, and provides valuable and objective information required in selecting and adjusting therapeutic regimens for the treatment of fungal keratitis.     

共聚焦显微镜在真菌性角膜炎中的应用

目的:探讨共聚焦显微镜在真菌性角膜炎中的应用价值。方法:回顾性分析在我院就诊的拟诊为真菌性角膜炎患者42例42眼,行共聚焦显微镜检查,同时行角膜病灶组织刮片染色检查。结果:真菌性角膜炎患者42例42眼,行共聚焦显微镜检查,观察到真菌菌丝为39例39眼,检出阳性率93%(39/42),行角膜病灶组织刮片染色镜检,观察到真菌菌丝为30例30眼,检出阳性率71%(30/42)。统计学分析表明两者差异有统计学意义(χ2=6.574,P<0.05)。结论:共聚焦显微镜是一种无创、高效、直观的检查设备,对真菌性角膜炎的初步诊断有着重要的临床价值。     

In vivo confocal microscopy in the diagnosis of fungal keratitis: case report

The authors describe a case of fungal keratitis that the in vivo confocal microscopy helped in the diagnosis and follow-up. Confocal microscopy was done in a patient's ulcer that did not improve with several topical medicines. Corneal scrapings were obtained and culture results were without conclusion. We observed hyphae and infectious collections on confocal microscopy. New corneal culture showed Fusarium sp ten days after confocal diagnosis.     

光学性全厚板层角膜移植术治疗真菌性角膜炎的初步探讨

目的初步探讨完全钝性分离的光学性全厚板层角膜移植术治疗真菌性角膜炎的可行性、安全性和增视效果。方法采用前瞻性非对照研究,对临床确诊为真菌性角膜炎且常规抗真菌药物治疗无效的患者行活体激光共焦显微镜检查．对其中明确菌丝未累及角膜深基质及后弹力层的21例患者（21眼）行完全钝性分离的光学性全厚板层角膜移植术。术后第1周、第1个月、第3个月、第6个月、第12个月和第18个月常规随访,观察角膜植片上皮愈合情况、有无真菌复发、免疫排斥反应、移植片透明度和最佳矫正视力等;术后第6个月和第12个月复查角膜内皮细胞密度。结果术前激光共焦显微镜检查。21只患眼均找到真菌菌丝,未见菌丝累及角膜深基质及后弹力层：术中无一例因后弹力层穿破而改行穿透性角膜移植术;仅1例在术后2周内出现真菌复发,复发率为4．76％。其余20例平均随访（14±6）个月,角膜植片均透明,治愈率为95．24％。术后第6个月,有14例术眼最佳矫正视力≥4．5,其中5例≥4．8。术后第6个月,平均角膜内皮细胞密度为（2296±368）个／mm^2（1025～3491个／mm^2）;术后第12个月为（2337±357）个／mm^2（1016～3380个／mm^2）。结论光学性全厚板层角膜移植术是一种治疗真菌性角膜炎的有效手段．激光共焦显微镜有助于真菌的诊断和真菌浸润深度的判断．完全钝性分离的方法减少了前房穿孔率,提高了手术的成功率。     

真菌性角膜炎70例早期临床诊断分析

目的:分析70例真菌性角膜炎早期临床诊断结果,评价共焦激光显微镜在真菌性角膜炎早期临床诊断中的应用价值。方法:用共焦激光角膜显微镜对临床拟诊为真菌性角膜炎的70例患者进行检查,同时行涂片检查真菌菌丝及培养查真菌菌落,对三组结果进行比较。结果:在70例患者中,根据病史、临床表现确诊为真菌性角膜炎,其中共焦激光角膜显微镜的确诊率为94%(66/70),角膜刮片的确诊率为31%(22/70),培养查真菌菌落确诊率为20%(14/70)。激光共焦角膜显微镜检查检出率高于角膜组织刮片(χ2=29.615,P<0.01)及真菌培养(χ2=39.433,P<0.01)检查。结论:共焦激光角膜显微镜是一种无创、快速、有效的活体检查方法,在真菌性角膜炎的早期诊断、治疗和研究中将起重要作用。     

Fungal keratitis caused by Cylindrocarpon lichenicola

We report a case of a 67-year-old woman with no significant past ocular history, who was referred for management of an unresponsive microbial keratitis resulting from trauma with a piece of clothing fabric 1 month previously in Portugal and worsening despite topical fortified antibiotics. On examination, visual acuity was limited to "light perception". Slit lamp examination revealed an 11×11mm full-thickness corneal infiltrate. Confocal images showed branching hyphae suggestive of a fungal infection. Fungal cultures of corneal scrapings revealed growth of Cylindrocarpon lichenicola, a saprophytic, filamentous fungus, which is an unusual cause of keratitis. Despite aggressive antifungal therapy with voriconazole and amphotericin B, she required penetrating keratoplasty for impending corneal perforation. Follow-up was uneventful, with no recurrence at 1 year. Fungal infections must be suspected in all corneal ulcers of traumatic etiology. Specific cultures and confocal microscopy must be performed early, so as to enable early treatment modification.     

Lamellar keratoplasty for the treatment of fungal keratitis.

PURPOSE: To determine the therapeutic value of lamellar keratoplasty (LKP) in the treatment of fungal keratitis not curable by antifungal chemotherapy. METHODS: Fifty-five patients, in whom a diagnosis of fungal keratitis was confirmed by microscopic analysis of corneal scrapings or confocal microscopy, and who were not cured by topical and oral antifungal medication, were given LKP. After LKP, topical antifungal treatment was continued for 2 weeks with gradual tapering of the drugs. The excised recipient lamella was used for microbial culture and histopathologic examination. RESULTS: Therapeutically beneficial results were achieved in 51 cases (92.7%) of the 55 LKPs that were performed. In these 51 cases, there was no recurrence of infection, and the resulting visual acuity ranged from 20/63 to 20/20. Patient follow-up ranged from 6 to 18 months. In four cases (7.3%), there was a recurrence of the fungal infection within 2 weeks of LKP. In these four patients, the infection was cured by performing a penetrating keratoplasty (PKP). Forty-six of the recipient lamellae were culture positive for fungi. Thirty-three of these cultures were identified as Fusarium, six as Aspergillus, three as Candida, one as Penicillium species, and in the other three cases, unidentified septate hyphae were noted. In the four cases of recurrent infection, microbiologic culture revealed three cases with Fusarium species and one case with Aspergillus species. Histopathologic analysis of periodic acid-Schiff (PAS)-stained tissue sections of donor lamellae revealed fungal filaments in all samples. Immune reactions to the lamellar grafts were not observed and the donor lamellae remained clear for the duration of follow-up. CONCLUSION: Lamellar keratoplasty can be effective for treating fungal keratitis that is not cured by antifungal therapeutics. In addition, LKP can provide useful vision with few complications. Furthermore, corneal tissue used in LKP may be obtained more easily than healthy tissue used in PKP.     

Clinical observation of removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis

AIM: To study the clinical observation of removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis. METHODS：A retrospective study was done to 10 patients (10 eyes) who had accepted removal of the necrotic corneal tissue combined with conjunctival flap covering surgery for fungal keratitis,the diagnosis by corneal scraping and smear examination or confocal microscopy check hyphae.Local and systemic antifungal therapy more than one week for all patients, corneal ulcer enlarge or no shrink.Slit lamp microscope examination the diameter of corneal ulcer about 2mm-4mm.Anterior segment optical coherence tomography（AS-OCT）examine the depth of corneal ulcer between 1/3-1/2,infiltrate corneal stroma about 20um-80um,the diameter of corneal ulcer about 3mm-6mm.Type-B ultrasonic exclusion endophthalmitis. Complete removal lesions until transparent of stroma, make conjunctival flap equal or greater than ulcer 1mm nearby conjunctiva. Continued antifungal therapy. The vision, fungal recurrence, conjunctival flap rollback or desquamate were analysed. RESULTS：Ten patients had success done this surgery, the corneal ulcer was not enlarge and healing afteroperation.7 cases were bridging conjunctival flap and 3cases were single conjunctival flap. Preoperation vision above 0.1 had 8 cases,7 cases had vision above 0.1 one week after surgery, while 1 cases vision droped from 0.3 to 0.05.There was not recurrent for fungal,2 cases conjunctival flap rollback:1 case was bridging and 1case was single flap, no conjunctival flap desquamate. CONCLUSION：It is safe and effective to perform removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis which werenot sensitive or aggravate for antifungal drugs.     

Treatment of fungal keratitis by penetrating keratoplasty

AIM: To study the use of penetrating keratoplasty (PKP) for the treatment of severe fungal keratitis that could not be cured by antifungal medication. METHODS: A retrospective analysis of 108 cases of severe fungal keratitis in which PKP was performed served as the basis for this study. Fungal keratitis was diagnosed by KOH staining of corneal scrapings or by confocal microscopic imaging of the cornea. All patients received a combination of topical and oral antifungal medicines without steroids as the first course of therapy. Patients whose corneal infection was not cured or in whom the infection progressed during antifungal treatment were given a PKP. After surgery, the patients continued to receive antifungal therapy with gradual tapering of the dose over a 1-2 month period. Cyclosporine was used to prevent graft rejection beginning 2 weeks after PKP. Topical steroid only was administered to the patient whose donor graft was over 8.5 mm and with a heavy iris inflammation 2 weeks after PKP. The surgical specimens were used for microbiological evaluation and examined histopathologically. The patients were followed for 6-24 months after PKP. Graft rejection, clarity of the graft, visual acuity, and surgical complications were recorded. RESULTS: Corneal grafts in 86 eyes (79.6%) remained clear during follow up. There was no recurrence of fungal infection and the visual acuity ranged from 40/200 to 20/20. Complications in some patients included recurrent fungal infection in eight eyes (7.4%), corneal graft rejection in 32 eyes (29.6%), secondary glaucoma in two eyes (1.9%), and five eyes (4.6%) developed cataracts. 98 of 108 of the recipient corneas had PAS positive fungal hyphae in tissue sections; 97 of 108 were culture positive for various fungi, including Fusarium (63), Aspergillus (14), Candida (9), Penicillium (4), and seven cases in which septate hyphae were seen but identification of the organism was not conclusive. CONCLUSIONS: PKP is an effective treatment for fungal keratitis that does not respond to antifungal medication. Early surgical intervention before the disease becomes advanced is recommended. It is critical that the surgical procedure remove the infected tissue in its entirety in order to effect a cure.