Q-开关Nd:YAG激光治疗真菌性角膜溃疡的实验研究
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摘要
目的：探讨Q-开关Nd:YAG激光治疗真菌性角膜溃疡的可行性及筛选最佳治疗参数，观察治疗效果。
方法
1 接种剂型制备
标准的串珠镰刀菌菌种实验前接种到含有新鲜配置的沙保弱葡萄糖琼脂培养基的培养皿中，28℃培养6天。培养的串珠镰刀菌用研磨器研磨成小的菌丝段，生理盐水稀释，红细胞计数板计数，配成浓度为5×107孢子或菌丝段的悬浮液用于接种。
2 实验动物与分组
新西兰白兔44只，体重2.0~2.5kg，雌雄不限，除外眼部疾患。第一步用24只，随机分为5组，第一组为正常对照组（2只兔4只眼），第二组为未治疗组（4只兔8只眼），另三组为激光治疗组（每组6只兔12只眼），激光能量分别为5mJ、10mJ、20mJ；第二步用20只，随机分为2组，每组10只兔20只眼，双眼接种，一组为激光加药物治疗组，另一组为局部清创加药物治疗组。
3 动物模型的制备
实验用兔除正常对照组外，均双眼接种串珠镰刀菌。实验前3天，0.25%氯霉素滴眼液点双眼，4次/日。麻醉：用氯胺酮肌注（40mg/kg），1%丁卡因点眼。刮除角膜中央约3mm角膜上皮，1ml注射器针头划伤角膜浅层，滴上0.2ml
已经制好的串珠镰刀菌菌液，盖上角膜接触镜，将第三眼睑与对侧球结膜缝合，并间断缝合睑裂。24小时后打开睑裂，拆除第三眼睑缝线，去除角膜接触镜，观察角膜感染情况，感染不重者继续戴镜，缝合睑裂，待12~24小时后再打开。
4 实验方法
4.1第一步
4.1.1激光治疗：动物模型建立后第5天病变达中基质层时，实验动物于全麻后用Q-开关Nd:YAG激光：光斑500um,能量分别为5mJ、10mJ、20mJ，单或双脉冲，采用由外到内，由浅至深的方法，对病变表面坏死组织进行定点爆破式清除，并记录击打点数。
4.1.2观察指标
4.1.2.1一般情况观察：动物模型建立后每天用裂隙灯对角膜及前房反应等情况进行观察。
4.1.2.2角膜内皮染色：正常对照组、未治疗组、治疗后1小时、第1天不同能量组各摘除一眼，用锥蓝联合茜素红进行角膜内皮细胞染色，观察角膜内皮细胞变化。
4.1.2.3病理切片光镜观察：正常对照组、未治疗组、治疗后第1天、第3天、第7天不同能量组各摘除一眼，常规石蜡切片，分别做HE及PAS染色，观察角膜的组织结构变化。
4.1.2.4透射电镜观察：正常对照组、未治疗组、治疗后1小时不同能量组各摘除一眼，透射电镜观察角膜内皮细胞的超微组织结构变化。
4.2第二步
4.2.1选用第一步中对角膜损伤较小且效果较好的激光参数对实验组进行治疗，同时每日球结膜下注射2%的氟康唑注
射液0.5ml；对照组用刀片进行局部清创，同时每日球结膜下注射2%的氟康唑注射液0.5ml。
4.2.2每日对兔眼角膜行裂隙灯检查观察角膜及前房反应情况，共7天。比较两组治愈率。
结果
1 动物模型
用浅层划痕，戴软性角膜接触镜联合第三眼睑缝合的方法制成的真菌性角膜溃疡动物模型100%成功，且获得的多为轻中度真菌性角膜溃疡模型。
2 内皮细胞染色
未治疗组及5mJ激光治疗组治疗后1小时角膜内皮细胞排列稍紊乱，细胞体积变大；10mJ激光治疗组治疗后1小时角膜内皮细胞排列紊乱，细胞体积变大，形状不规则，边界模糊，细胞间隙增宽；20mJ激光治疗组治疗后1小时角膜内皮细胞排列明显紊乱，细胞形状明显不规则，边界模糊，细胞间隙明显增宽，可见细胞缺失；术后第1天各激光治疗组角膜内皮细胞可见排列紊乱，细胞形状不规则，未见细胞缺失，未见核着色，各组细胞间隙较前变窄，余未见明显区别。
3 病理
未治疗组HE染色主要病理变化为广泛的化脓性炎症，PAS染色可见实质层内有淡粉红色的真菌菌丝生长；5mJ组HE染色实质层有轻度水肿，3天后溃疡面愈合，但炎症在实质层及后部继续发展，7天后形成角膜后脓肿，PAS染色治疗后第3天即见真菌菌丝；10mJ组HE染色实质层中度水肿，3天后炎症反应减轻，7天后炎症基本消失，上皮增生，结缔组织增生，角膜斑翳形成，PAS染色未见真菌菌丝；20mJ
组HE染色实质层水肿严重，7天后仍见实质层明显水肿，PAS染色未见真菌菌丝。
4 透射电镜
未治疗组与5mJ组可见角膜内皮细胞基质轻度水肿，细胞内线粒体部分嵴消失；10mJ组可见角膜内皮细胞基质中度水肿，细胞内线粒体呈空泡样变，嵴大部分消失；20mJ组可见角膜内皮细胞破裂，细胞基质严重水肿，线粒体呈空泡样变，嵴大部分消失。
5 各不同能量激光治疗组击打点数
5mJ组与10mJ组，5mJ组与20mJ组比较，P＜0.05，有统计学差异。10mJ组与20mJ组比较，P＞0.05，无统计学差异。
6 用10mJ Q-开关Nd:YAG激光加药物治疗真菌性角膜溃疡治愈率可达80%，而局部清创加药物治愈率为40%，二者进行校正X2检验，P＜0.05，具有统计学差异。
结论
1、能量为10mJ组对角膜组织损伤为可逆性，且能较彻底清除病变组织，为治疗真菌性角膜溃疡的一种有效而简便的新方法。
2、能量为10mJ联合抗真菌药物治疗组真菌性角膜溃疡，治愈率达80%，高于局部清创加药物治疗组。
3、Q-开关Nd:YAG激光有可能成为一种治疗真菌性角膜溃疡的新方法。
Objective：To evaluate the possibility of treatment of fungal corneal ulcer using Q-switched Nd:YAG laser. To investgate the best treatment parameters and observe the therapeutic effects of Q-switched Nd:YAG laser for fungal corneal ulcer.
Methods
1 inoculaed
The Fusarium specie was inoculated on Sabouraud,s agar, and grown six days at 28℃. Then the fungal specie was crushed, reconstituted in sterile water, counted on a hemocytometer, and the released Fusarium adjusted to 5×107/ml .
Animals
The first step：24 healthy New Zealand white rabbits weighing 2.0~2.5Kg were divided randomly into 5 groups, one was normal group（2rabbits, 4eyes）. one was experimental group(4rabbits, 8eyes). the other three were control groups(6 rabbits 12eyes each). Energy was 5mJ、10mJ、20mJ repectively. The second step：20 rabbits were divided randomly into 2 groups, (10 rabbits, 20eyes each). One group was treated by laser combining medicine, the other group was treated by debridement combining medicine.
Making of animal model
Double eyes were inoculaed in every rabbit. 0.25% chloromycetin was applied topically 3days before inoculated. The rabbits were anesthesized. The central corneal epithelium was removed within a 3mm diameter and 0.2ml Fusarium was instilled on the corneal surface. A soft contact lens was used to cover the cornea. Then sutured the third eyelids with the opposite bulbar conjunctiva and tarsorrhaphy was performed, took out stitches 24 hours post-inoculation, examined the cornea. If the infection was not severe, Tarsorrhaphy was performed again, took out stitches 12 or 24 hours later again.
4 Experiment method
4.1 The first step
4.1.1 Laser treatment：The pathological changes has reached the middle of lamellae 5 days post-inoculation. The necrosis on the surface of cornea ulcer was debrided by pinpoint bombing uaing Q-switched Nd:YAG laser with an output energy of 5mJ、10mJ、20mJ respectively, spot 500um in diameter from outer to center, and from the surface to the deep until the surface of ulcer become a clean one, noting the number of treating at the same time.
Observe target
Clinical examination：Rabbits were examined by slit-lamp biomicroscopy everyday.
Stained corneal endothelial cell：The corneal endothelial cell were stained using alizarin red S-trypan blue in
normal group、control group、1hour and 1 day post-laser respectively. Observed the corneal endothelial cell.
Light microscopy：Histological changes were examined in normal group、control group、1hour and 1 day post-laser respectively.
Electron transmicroscopy：The corneal endothelial cell ultrastructure changes were examined in normal group、control group、1hour and 1 day post-laser respectively.
The second step
4.2.1 The treated group animals were treated using the laser that was selected at the first step. 0.5ml 0.2% Fluconazole was injected subconjunctival everyday after laser treatment. The control group animal were treated using corneal lesion resection and 0.5ml 0.2% Fluconazole was injected subconjunctival everyday.
4.2.2 Clinical examination：Animals were examined by slit-lamp biomicroscopy everyday, compared the cure rates of the two groups.
Results
Making of animal model
Nicking the surface layer, using the soft contact lens combined with performing the tarsorrhaphy at the same time was a very good method of making the fungal corneal ulcer animal model. All the animal models were succeeded. Most of the infection eyes were in light or middle degree.
Stained corneal endothelial cell
The control group and 1hour post-5mJ laser：There was a little of irregular in the shape of the endothelial cells. Cubage of the cells was enlarged; 1hour post-10mJ laser：The shape of the corneal endothelial cells was irregular obviously. The cubage of the corneal endothelial cells was enlarged. The boundary of the cells was not obvious; 1hour post-20mJ laser：The shape of some corneal endothelial cell was irregular obviously. Some corneal endothelial cells were lost; 1 day post-laser：the shape of endothelial cell of
方法
1 接种剂型制备
标准的串珠镰刀菌菌种实验前接种到含有新鲜配置的沙保弱葡萄糖琼脂培养基的培养皿中，28℃培养6天。培养的串珠镰刀菌用研磨器研磨成小的菌丝段，生理盐水稀释，红细胞计数板计数，配成浓度为5×107孢子或菌丝段的悬浮液用于接种。
2 实验动物与分组
新西兰白兔44只，体重2.0~2.5kg，雌雄不限，除外眼部疾患。第一步用24只，随机分为5组，第一组为正常对照组（2只兔4只眼），第二组为未治疗组（4只兔8只眼），另三组为激光治疗组（每组6只兔12只眼），激光能量分别为5mJ、10mJ、20mJ；第二步用20只，随机分为2组，每组10只兔20只眼，双眼接种，一组为激光加药物治疗组，另一组为局部清创加药物治疗组。
3 动物模型的制备
实验用兔除正常对照组外，均双眼接种串珠镰刀菌。实验前3天，0.25%氯霉素滴眼液点双眼，4次/日。麻醉：用氯胺酮肌注（40mg/kg），1%丁卡因点眼。刮除角膜中央约3mm角膜上皮，1ml注射器针头划伤角膜浅层，滴上0.2ml
已经制好的串珠镰刀菌菌液，盖上角膜接触镜，将第三眼睑与对侧球结膜缝合，并间断缝合睑裂。24小时后打开睑裂，拆除第三眼睑缝线，去除角膜接触镜，观察角膜感染情况，感染不重者继续戴镜，缝合睑裂，待12~24小时后再打开。
4 实验方法
4.1第一步
4.1.1激光治疗：动物模型建立后第5天病变达中基质层时，实验动物于全麻后用Q-开关Nd:YAG激光：光斑500um,能量分别为5mJ、10mJ、20mJ，单或双脉冲，采用由外到内，由浅至深的方法，对病变表面坏死组织进行定点爆破式清除，并记录击打点数。
4.1.2观察指标
4.1.2.1一般情况观察：动物模型建立后每天用裂隙灯对角膜及前房反应等情况进行观察。
4.1.2.2角膜内皮染色：正常对照组、未治疗组、治疗后1小时、第1天不同能量组各摘除一眼，用锥蓝联合茜素红进行角膜内皮细胞染色，观察角膜内皮细胞变化。
4.1.2.3病理切片光镜观察：正常对照组、未治疗组、治疗后第1天、第3天、第7天不同能量组各摘除一眼，常规石蜡切片，分别做HE及PAS染色，观察角膜的组织结构变化。
4.1.2.4透射电镜观察：正常对照组、未治疗组、治疗后1小时不同能量组各摘除一眼，透射电镜观察角膜内皮细胞的超微组织结构变化。
4.2第二步
4.2.1选用第一步中对角膜损伤较小且效果较好的激光参数对实验组进行治疗，同时每日球结膜下注射2%的氟康唑注
射液0.5ml；对照组用刀片进行局部清创，同时每日球结膜下注射2%的氟康唑注射液0.5ml。
4.2.2每日对兔眼角膜行裂隙灯检查观察角膜及前房反应情况，共7天。比较两组治愈率。
结果
1 动物模型
用浅层划痕，戴软性角膜接触镜联合第三眼睑缝合的方法制成的真菌性角膜溃疡动物模型100%成功，且获得的多为轻中度真菌性角膜溃疡模型。
2 内皮细胞染色
未治疗组及5mJ激光治疗组治疗后1小时角膜内皮细胞排列稍紊乱，细胞体积变大；10mJ激光治疗组治疗后1小时角膜内皮细胞排列紊乱，细胞体积变大，形状不规则，边界模糊，细胞间隙增宽；20mJ激光治疗组治疗后1小时角膜内皮细胞排列明显紊乱，细胞形状明显不规则，边界模糊，细胞间隙明显增宽，可见细胞缺失；术后第1天各激光治疗组角膜内皮细胞可见排列紊乱，细胞形状不规则，未见细胞缺失，未见核着色，各组细胞间隙较前变窄，余未见明显区别。
3 病理
未治疗组HE染色主要病理变化为广泛的化脓性炎症，PAS染色可见实质层内有淡粉红色的真菌菌丝生长；5mJ组HE染色实质层有轻度水肿，3天后溃疡面愈合，但炎症在实质层及后部继续发展，7天后形成角膜后脓肿，PAS染色治疗后第3天即见真菌菌丝；10mJ组HE染色实质层中度水肿，3天后炎症反应减轻，7天后炎症基本消失，上皮增生，结缔组织增生，角膜斑翳形成，PAS染色未见真菌菌丝；20mJ
组HE染色实质层水肿严重，7天后仍见实质层明显水肿，PAS染色未见真菌菌丝。
4 透射电镜
未治疗组与5mJ组可见角膜内皮细胞基质轻度水肿，细胞内线粒体部分嵴消失；10mJ组可见角膜内皮细胞基质中度水肿，细胞内线粒体呈空泡样变，嵴大部分消失；20mJ组可见角膜内皮细胞破裂，细胞基质严重水肿，线粒体呈空泡样变，嵴大部分消失。
5 各不同能量激光治疗组击打点数
5mJ组与10mJ组，5mJ组与20mJ组比较，P＜0.05，有统计学差异。10mJ组与20mJ组比较，P＞0.05，无统计学差异。
6 用10mJ Q-开关Nd:YAG激光加药物治疗真菌性角膜溃疡治愈率可达80%，而局部清创加药物治愈率为40%，二者进行校正X2检验，P＜0.05，具有统计学差异。
结论
1、能量为10mJ组对角膜组织损伤为可逆性，且能较彻底清除病变组织，为治疗真菌性角膜溃疡的一种有效而简便的新方法。
2、能量为10mJ联合抗真菌药物治疗组真菌性角膜溃疡，治愈率达80%，高于局部清创加药物治疗组。
3、Q-开关Nd:YAG激光有可能成为一种治疗真菌性角膜溃疡的新方法。
Objective：To evaluate the possibility of treatment of fungal corneal ulcer using Q-switched Nd:YAG laser. To investgate the best treatment parameters and observe the therapeutic effects of Q-switched Nd:YAG laser for fungal corneal ulcer.
Methods
1 inoculaed
The Fusarium specie was inoculated on Sabouraud,s agar, and grown six days at 28℃. Then the fungal specie was crushed, reconstituted in sterile water, counted on a hemocytometer, and the released Fusarium adjusted to 5×107/ml .
Animals
The first step：24 healthy New Zealand white rabbits weighing 2.0~2.5Kg were divided randomly into 5 groups, one was normal group（2rabbits, 4eyes）. one was experimental group(4rabbits, 8eyes). the other three were control groups(6 rabbits 12eyes each). Energy was 5mJ、10mJ、20mJ repectively. The second step：20 rabbits were divided randomly into 2 groups, (10 rabbits, 20eyes each). One group was treated by laser combining medicine, the other group was treated by debridement combining medicine.
Making of animal model
Double eyes were inoculaed in every rabbit. 0.25% chloromycetin was applied topically 3days before inoculated. The rabbits were anesthesized. The central corneal epithelium was removed within a 3mm diameter and 0.2ml Fusarium was instilled on the corneal surface. A soft contact lens was used to cover the cornea. Then sutured the third eyelids with the opposite bulbar conjunctiva and tarsorrhaphy was performed, took out stitches 24 hours post-inoculation, examined the cornea. If the infection was not severe, Tarsorrhaphy was performed again, took out stitches 12 or 24 hours later again.
4 Experiment method
4.1 The first step
4.1.1 Laser treatment：The pathological changes has reached the middle of lamellae 5 days post-inoculation. The necrosis on the surface of cornea ulcer was debrided by pinpoint bombing uaing Q-switched Nd:YAG laser with an output energy of 5mJ、10mJ、20mJ respectively, spot 500um in diameter from outer to center, and from the surface to the deep until the surface of ulcer become a clean one, noting the number of treating at the same time.
Observe target
Clinical examination：Rabbits were examined by slit-lamp biomicroscopy everyday.
Stained corneal endothelial cell：The corneal endothelial cell were stained using alizarin red S-trypan blue in
normal group、control group、1hour and 1 day post-laser respectively. Observed the corneal endothelial cell.
Light microscopy：Histological changes were examined in normal group、control group、1hour and 1 day post-laser respectively.
Electron transmicroscopy：The corneal endothelial cell ultrastructure changes were examined in normal group、control group、1hour and 1 day post-laser respectively.
The second step
4.2.1 The treated group animals were treated using the laser that was selected at the first step. 0.5ml 0.2% Fluconazole was injected subconjunctival everyday after laser treatment. The control group animal were treated using corneal lesion resection and 0.5ml 0.2% Fluconazole was injected subconjunctival everyday.
4.2.2 Clinical examination：Animals were examined by slit-lamp biomicroscopy everyday, compared the cure rates of the two groups.
Results
Making of animal model
Nicking the surface layer, using the soft contact lens combined with performing the tarsorrhaphy at the same time was a very good method of making the fungal corneal ulcer animal model. All the animal models were succeeded. Most of the infection eyes were in light or middle degree.
Stained corneal endothelial cell
The control group and 1hour post-5mJ laser：There was a little of irregular in the shape of the endothelial cells. Cubage of the cells was enlarged; 1hour post-10mJ laser：The shape of the corneal endothelial cells was irregular obviously. The cubage of the corneal endothelial cells was enlarged. The boundary of the cells was not obvious; 1hour post-20mJ laser：The shape of some corneal endothelial cell was irregular obviously. Some corneal endothelial cells were lost; 1 day post-laser：the shape of endothelial cell of
引文
卢嘉彪，陈家祺，王丽娅，广东地区真菌性角膜病的病原体及其发病情况的变迁，眼科研究，1998，16：289~291。
王丽娅，王莉，张月琴等，真菌性角膜病在河南地区的流行病学研究，中国全科医学杂志，1998，1：152~154。
张士元，新中国眼科50年，中华眼科杂志，1999，35：325~329
Killingsworth DW, Stern GA, Driebe WT, et al Results of therapeutic penetrating keratoplasty. Ophthalmology, 1993, 100:534~541
李建国，杨玉蓉，崔国华等，Nd:YAG激光在真菌性角膜溃疡中的应用，中国激光医学杂志，2000，4：207
马吉献，张廷钺，于纯智等，用茜素红-锥蓝联合染色法对兔角膜内皮细胞活性的评价，眼科研究，1993，11：1~3
O,Day DM, Ray WA, Head WS, et al Influence of corticosteroid on experimental induced keratomycosis. ArchOpthalmology. 1991, 109:1601~1604
Forster RK, Rebell G, Animal model of Fusarium Solani keratitis. Am J Ophthal, 1975, 79:510~516
O,Day DM, Head WS, Robinson RD, et al The evaluation of therapeutic responses in experimental keratomycosis, Curr Eye Res. 1992, 11:35~44
Yamamoto GK, Pavan-Langston D, Stowe GC, et al Fungal invasion of a therapeutic soft contact lens and cornea. Ann of Ophthal. 1979, 11:1731~1735
Churner R, Cunningham RD, Fungal contaminated soft contact lenses. Ann Ophthal, 1983, 15:724~727
Kremer I, Goldenfeld M, Snmueli D. Fungal keratitis associated with contact lens wear after penetrating keratoplasty, Ann Ophthal, 1991, 23:342~345
Ritterband DC, Seedor JA, Shah MK. et al A unique case of cryptococcus laurentii keratitis spread by a rigid gas permeable contact lens in a patient with onychomycosis. Cornea, 1998, 17:115~118
朱志忠等，角膜病学，人民卫生出版社，第一版，1986：272~273
Stocker FW, King EH, Lucas DO, et al Clinical test for evaluating donor corneas, Arch Ophthalmol , 1970, 84:2~7
邱孝芝，蒋秀莉，蒋雪芹等，深低温保存角膜的实验研究，中华眼科杂志，1988，1：41~43
孙为荣等，眼科病理学，人民卫生出版社，第一版，1987，20
王新月，郭娟，孙声桃等，外伤所致真菌性角膜溃疡的病原学及治疗分析，眼外伤职业病杂志，2002，24：519~520
周祖濂综述，霉菌性角膜炎，国外医学·眼科学分册，1991，15：23~27
朱菁等，激光医学，上海科学技术出版社，第一版，2003，207
孟祥毓等，病灶切除治疗17例真菌性角膜炎，眼科，1992，1：48
王印其，高长凤，赵东卿等，角膜真菌病，中国实用眼科
杂志，1996，14：517~522
王丽娅，张月琴，王印其等，中国三地区真菌性角膜病致病菌种的调查，中华眼科杂志，2000，36：138~140
王丽娅，王莉，张月琴等，真菌性角膜病在河南地区的流行病学研究，中国全科医学杂志，1998，1：152~154。
张士元，新中国眼科50年，中华眼科杂志，1999，35：325~329
Killingsworth DW, Stern GA, Driebe WT, et al Results of therapeutic penetrating keratoplasty. Ophthalmology, 1993, 100:534~541
李建国，杨玉蓉，崔国华等，Nd:YAG激光在真菌性角膜溃疡中的应用，中国激光医学杂志，2000，4：207
马吉献，张廷钺，于纯智等，用茜素红-锥蓝联合染色法对兔角膜内皮细胞活性的评价，眼科研究，1993，11：1~3
O,Day DM, Ray WA, Head WS, et al Influence of corticosteroid on experimental induced keratomycosis. ArchOpthalmology. 1991, 109:1601~1604
Forster RK, Rebell G, Animal model of Fusarium Solani keratitis. Am J Ophthal, 1975, 79:510~516
O,Day DM, Head WS, Robinson RD, et al The evaluation of therapeutic responses in experimental keratomycosis, Curr Eye Res. 1992, 11:35~44
Yamamoto GK, Pavan-Langston D, Stowe GC, et al Fungal invasion of a therapeutic soft contact lens and cornea. Ann of Ophthal. 1979, 11:1731~1735
Churner R, Cunningham RD, Fungal contaminated soft contact lenses. Ann Ophthal, 1983, 15:724~727
Kremer I, Goldenfeld M, Snmueli D. Fungal keratitis associated with contact lens wear after penetrating keratoplasty, Ann Ophthal, 1991, 23:342~345
Ritterband DC, Seedor JA, Shah MK. et al A unique case of cryptococcus laurentii keratitis spread by a rigid gas permeable contact lens in a patient with onychomycosis. Cornea, 1998, 17:115~118
朱志忠等，角膜病学，人民卫生出版社，第一版，1986：272~273
Stocker FW, King EH, Lucas DO, et al Clinical test for evaluating donor corneas, Arch Ophthalmol , 1970, 84:2~7
邱孝芝，蒋秀莉，蒋雪芹等，深低温保存角膜的实验研究，中华眼科杂志，1988，1：41~43
孙为荣等，眼科病理学，人民卫生出版社，第一版，1987，20
王新月，郭娟，孙声桃等，外伤所致真菌性角膜溃疡的病原学及治疗分析，眼外伤职业病杂志，2002，24：519~520
周祖濂综述，霉菌性角膜炎，国外医学·眼科学分册，1991，15：23~27
朱菁等，激光医学，上海科学技术出版社，第一版，2003，207
孟祥毓等，病灶切除治疗17例真菌性角膜炎，眼科，1992，1：48
王印其，高长凤，赵东卿等，角膜真菌病，中国实用眼科
杂志，1996，14：517~522
王丽娅，张月琴，王印其等，中国三地区真菌性角膜病致病菌种的调查，中华眼科杂志，2000，36：138~140