玻璃体腔重复注射抗血管内皮生长因子单克隆抗体bevacizumab对糖尿病大鼠视网膜的毒性观察

Objective To observe the retinal toxicity of repeated intravitreal injection with bevacizumab(Avastin)in diabetic rats.Methods Forty male Sprague Dawley(SD)rats were randomly divided into normal group(Group A,10 rats)and diabetes mellitus group(30 rats).The rats in diabetes mellitus group were induced with streptozotocin injection for diabetic retinopathy model.And then randomly divided into diabetic retinopathy(DR)group(Group B,10 rats),the rats were not intervened;the left eyes of the other 20 rats were intravitreal injected with bevaeizumab 3 μ1(25 mg/m1)for 3 times as experimental group(Group C);the right eyes of the 20 rats were not intervened as experimental control group(Group D),20 days after last intravitreal injection,retinal function was measured by Flicker Electroretinogram (F-ERG);retinal vascular pattern was determined by fluorescence microscopy of ethidium bromide(EB)stained retinal flat mounts;retinal morphological changes were determined by light microscope on hematoxylin-eosin (HE) stained sections;Thy-1 and VEGF expression was measured by immunohistochemistry staining.Results F-ERG showed that-the differences of a-and b-waves-the b-wave amplitude and the Ops-wave amplitude in the implicit time between group A,B,C and D were significant (F=33.165,36.162,19.955,23.243;P=0.000);the differences of a-wave amplitude between group A,B,C and D was not significant(F=0.097,P=0v961).Retinal blood vessel pattern was normalin Group A;retinal vascular vessels were tortuous and irregularly expanded in Group B:retinal vascular vessels of Group C were regular and thinner than Group A;microaneurysm were showed in Group D.Light microscope displayed that the layers of the rat retina of Group A were regular,the retinal architectures of Group B were irregular,the retinal layers were regular in Group C,the retinal layers were irregular in Group D.Immunohistochemistry staining discovered that Thy-1 and VEGF were mainly expressed in ganglion cell layer(GCL).Conclusion Repeated intravitreal injection of bevacizumab iS toxic tO retina of diabetes mellitus rats.     

玻璃体腔重复注射抗血管内皮生长因子单克隆抗体bevacizumab对糖尿病大鼠视网膜的毒性观察

目的 观察玻璃体腔重复注射抗血管内皮生长因子(VEGF)单克隆抗体bevacizumab(商品名Avastin)对糖尿病大鼠视网膜的毒性作用.方法 40只健康成年雄性Sprague-Dawley大鼠随机分为正常组(A组)和糖尿病组,分别为10、30只.糖尿病组采用链脲佐菌霉素(STZ)尾静脉注射方法制作糖尿病动物模型.随机选取10只作为糖尿病视网膜病变(DR)组(B组),不作任何处理;其余20只大鼠左眼为实验组(C组),玻璃体腔注射25 mg/ml的bevacizumab 3 μ1,共注射3次,每次间隔10 d;右眼为实验对照组(D组),不给予任何处理.末次注射后20 d,采用闪光视网膜电图(F-ERG)对各组大鼠行视网膜功能检测;溴化乙锭(EB)染色视网膜铺片,荧光显微镜观察各组大鼠视网膜血管变化情况;苏木精-伊红(HE)染色,光学显微镜观察大鼠视网膜形态学变化;免疫组织化学染色法观察大鼠视网膜各层Thy-1及VEGF阳性表达情况.结果 F-ERG检测显示,A、B、C、D 4组暗适应a、b波潜伏期,暗适应b波振幅及振荡电位(Ops)总振幅比较,差异均有统计学意义(F=33.165,36.162,19.955,23.243;P值均=0.000);A、B、C、D4组暗适应a波振幅比较,差异无统计学意义(F=0.097,P=0.961).荧光显微镜观察发现,A组大鼠视网膜血管走行良好,B组大鼠视网膜血管走行纡曲、扩张,C组大鼠视网膜血管走形规则、变细,D组大鼠视网膜可见微血管瘤.光学显微镜观察发现,A组大鼠视网膜层次结构整齐分明,B组大鼠视网膜各层细胞结构排列紊乱,C组大鼠视网膜各层细胞排列整齐,D组大鼠视网膜各层细胞排列不整齐.免疫组织化学染色发现,Thy-1阳性表达主要位于神经节细胞层(GCL),极少数位于内丛状层、内核层.VEGF阳性表达主要定位于GCL,少量位于神经纤维层、内核层及视网膜色素上皮层.结论 玻璃体腔重复注射bevacizumab对糖尿病大鼠视网膜有一定毒性作用.

Abstract：

Objective To observe the retinal toxicity of repeated intravitreal injection with bevacizumab(Avastin)in diabetic rats.Methods Forty male Sprague Dawley(SD)rats were randomly divided into normal group(Group A,10 rats)and diabetes mellitus group(30 rats).The rats in diabetes mellitus group were induced with streptozotocin injection for diabetic retinopathy model.And then randomly divided into diabetic retinopathy(DR)group(Group B,10 rats),the rats were not intervened;the left eyes of the other 20 rats were intravitreal injected with bevaeizumab 3 μ1(25 mg/m1)for 3 times as experimental group(Group C);the right eyes of the 20 rats were not intervened as experimental control group(Group D),20 days after last intravitreal injection,retinal function was measured by Flicker Electroretinogram (F-ERG);retinal vascular pattern was determined by fluorescence microscopy of ethidium bromide(EB)stained retinal flat mounts;retinal morphological changes were determined by light microscope on hematoxylin-eosin (HE) stained sections;Thy-1 and VEGF expression was measured by immunohistochemistry staining.Results F-ERG showed that-the differences of a-and b-waves-the b-wave amplitude and the Ops-wave amplitude in the implicit time between group A,B,C and D were significant (F=33.165,36.162,19.955,23.243;P=0.000);the differences of a-wave amplitude between group A,B,C and D was not significant(F=0.097,P=0v961).Retinal blood vessel pattern was normalin Group A;retinal vascular vessels were tortuous and irregularly expanded in Group B:retinal vascular vessels of Group C were regular and thinner than Group A;microaneurysm were showed in Group D.Light microscope displayed that the layers of the rat retina of Group A were regular,the retinal architectures of Group B were irregular,the retinal layers were regular in Group C,the retinal layers were irregular in Group D.Immunohistochemistry staining discovered that Thy-1 and VEGF were mainly expressed in ganglion cell layer(GCL).Conclusion Repeated intravitreal injection of bevacizumab iS toxic tO retina of diabetes mellitus rats.     

鼠神经生长因子对早期糖尿病大鼠玻璃体中感光细胞间维生素A类结合蛋白含量的影响

Objective To observe the effect of intravitreal injection of mouse nerve growth factor (NGF)on interphotoreceptor retinoid-binding protein(IRBP)in the vitreous of diabetic rats at early stages.Methods Ninety-six male Sprague Dawley(SD)rats were divided into control group(group A,24 rats)and experimental group(72 rats).The rats in experimental group were induced with streptozotocin injection for diabetic retinopathy model,and then randomly divided into positive control group(group B),normal saline group(group C)and NGF group(group D),24 rats in each group.The rats in the group A and B were not intervened.The rats were received intravitreal injection with 4μl normal saline(group C)or 4 μl(0.5 μg/μ1)NGF(group D).At 2,4,6 and 8 weeks after injection,IRBP levels were detected bv enzvmelinked immunosorbent assay(ELISA);hematoxylin-eosin(HE)staining and light microscope were used to observe the morphological changes of the retina;transmission electron microscope was used to observe the retinal uhrastructure.Results At 2 weeks after injection,there was no significant difference in IRBP expression between group A,B,C and D(F=2.833,P=0.052).At 4,6,8 weeks after injection,the differences of IRBP expression between group A,B,C and D were significant(F=22.252,108.459,105.726;P=0.000).At different time points after injection,there was no significant difference in IRBP expression of group A(F=1.462,P=0.241),but there were significant differences in IRBP expression of group B.C and D(F=150.98,63.519,64.604;P=0.000).Light microscope found that the retinal structure was clear in group A and in group B,C,D at 2,4 weeks after injection;the retinal thickness were thinner in group B,C,D at 8 weeks after injection.Transmission electron microscope displayed that the structure of rod outer segments was clear in group A and in group B,C,D at 2 weeks after injection;partly unclear structure of rod outer segments and slightly enlarged gap were observed in group B,C,D at 4,8Weeks after injection.Conclusion Intravitreal injection with NGF can stabilize the IRBP expression in the vitreous of diabetic rats at early stages effectively.     

鼠神经生长因子对早期糖尿病大鼠玻璃体中感光细胞间维生素A类结合蛋白含量的影响

Objective To observe the effect of intravitreal injection of mouse nerve growth factor (NGF)on interphotoreceptor retinoid-binding protein(IRBP)in the vitreous of diabetic rats at early stages.Methods Ninety-six male Sprague Dawley(SD)rats were divided into control group(group A,24 rats)and experimental group(72 rats).The rats in experimental group were induced with streptozotocin injection for diabetic retinopathy model,and then randomly divided into positive control group(group B),normal saline group(group C)and NGF group(group D),24 rats in each group.The rats in the group A and B were not intervened.The rats were received intravitreal injection with 4μl normal saline(group C)or 4 μl(0.5 μg/μ1)NGF(group D).At 2,4,6 and 8 weeks after injection,IRBP levels were detected bv enzvmelinked immunosorbent assay(ELISA);hematoxylin-eosin(HE)staining and light microscope were used to observe the morphological changes of the retina;transmission electron microscope was used to observe the retinal uhrastructure.Results At 2 weeks after injection,there was no significant difference in IRBP expression between group A,B,C and D(F=2.833,P=0.052).At 4,6,8 weeks after injection,the differences of IRBP expression between group A,B,C and D were significant(F=22.252,108.459,105.726;P=0.000).At different time points after injection,there was no significant difference in IRBP expression of group A(F=1.462,P=0.241),but there were significant differences in IRBP expression of group B.C and D(F=150.98,63.519,64.604;P=0.000).Light microscope found that the retinal structure was clear in group A and in group B,C,D at 2,4 weeks after injection;the retinal thickness were thinner in group B,C,D at 8 weeks after injection.Transmission electron microscope displayed that the structure of rod outer segments was clear in group A and in group B,C,D at 2 weeks after injection;partly unclear structure of rod outer segments and slightly enlarged gap were observed in group B,C,D at 4,8Weeks after injection.Conclusion Intravitreal injection with NGF can stabilize the IRBP expression in the vitreous of diabetic rats at early stages effectively.     

超声微泡造影剂促进重组腺相关病毒载体介导基因转染视网膜神经节细胞的体内实验

Objective To investigate the enhancing effect of ultrasound microbubbles on transfection of recombinant adeno-associated virus (rAAV) mediated green fluorecent protein (EGFP) gene into retinal ganglion cells (RGC) in vivo. Methods A total of 40 adult Sprague-Dawley (SD) rats were divided into four groups randomly (group A, B, C, D) with 10 rats in each. Group A was the normal control, in which the rats underwent intravitreal injection with 5 μl phosphate buffered solution. The rats in group B underwent intravitreal injection with 5 μl recombinant adeno-associated virus encoding EGFP gene (rAAV2-EGFP). The rats in group C underwent ultrasound irradiation on eyes right after intravitreal injection with 5μl rAAV2-EGFP; The ultrasound irradiation was performed on the rats in group D right after intravitreal injection with the mixture solution of microbubbles and rAAV2-EGFP ultrasound. After 21 days, RGC were labeled retogradely with fluogold. Seven days after labeling, the retinal flatmounts and frozen sections were made from five rats in each group. Expression of EGFP reporter gene was observed by laser scanning confocal microscope and evaluated via average optical intensity (AOD) and RGC transfection rate. Labeled RGC were counted to evaluate the adverse effects. Results Green fluorescence can be observed exactly in labeled RGC in B,C, and D groups. The AOD and transfection rate in group D was (95.02 ± 7. 25)% and (20. 10±0. 74)% , respectively; which were higher than those in group B and C (F=25. 970,25. 799;P<0.01). The difference of the number of RGC among the four groups was not significant(F= 0. 877, P>0. 05). Conclusion Under the condition of low frequency and with certain energy, ultrasound-mediated microbubble destruction can effectively and safely enhance rAAV delivery to RGC in rats.     

鼠神经生长因子对早期糖尿病大鼠玻璃体中感光细胞间维生素A类结合蛋白含量的影响

目的 观察玻璃体腔注射鼠神经生长因子(NGF)对早期糖尿病大鼠玻璃体中感光细胞间维生素A类结合蛋白(IRBP)含量的影响.方法 Sprague-Dawley雄性大鼠96只,随机分为正常对照组(A 组)及实验组,分别为24、72只.实验组采用链脲佐菌霉素(STZ)一次性腹腔注射制作糖尿病动物模型,再随机分为糖尿病阳性对照组(B组)、糖尿病玻璃体腔生理盐水注射组(C组)和糖尿病玻璃体腔鼠NGF注射组(D组),每组24只大鼠.A组及B组建模后不给予任何干预;C组注射生理盐水4μl,1次/周;D组注射0.5μg/μl的鼠NGF 4 μl,1次/周.注射后2、4、6、8周,采用酶联免疫吸附测定(ELISA)法检测大鼠玻璃体中IRBP含量;作石蜡切片行苏木精-伊红(HE)染色,光学显微镜观察视网膜结构;作超微切片,透射电子显微镜观察视网膜超微结构.结果 注射后2周,A、B、C、D组大鼠玻璃体中IRBP含量比较,差异无统计学意义(F=2.833,P=0.052).注射后4、6、8周,A、B、C、D组大鼠玻璃体中IRBP含量比较,差异均有统计学意义(F=22.252,108.459,105.726;P值均=0.000).A组组内注射后不同时间点大鼠玻璃体中IRBP含量比较,差异无统计学意义(F=1.462,P=0.241);B、C、D组组内注射后不同时间点大鼠玻璃体中IRBP含量比较,差异均有统计学意义(F=150.98,63.519,64.604;P值均=0.000).光学显微镜观察发现,A组大鼠视网膜各层组织层次清楚,排列整齐.注射后2、4周,B、C、D组大鼠视网膜结构较A组无明显改变.注射后8周,B、C、D组大鼠视网膜厚度变薄.透射电子显微镜观察发现,A组大鼠视细胞外节膜盘结构清晰,排列规则.注射后2周,B、C、D组大鼠视细胞外节膜盘结构清晰,排列规则整齐.注射后4周,B、C、D组大鼠视细胞外节膜盘局部模糊不清,间隙略有扩大.注射后8周,B、C组大鼠视细胞外节膜盘局部模糊不清,间隙扩大,部分出现溶解、断裂;D组大鼠视细胞外节膜盘局部模糊不清,间隙扩大,未见明显溶解断裂.结论 玻璃体腔注射鼠NGF可以有效稳定糖尿病大鼠早期玻璃体中IRBP的含量.

Abstract：

Objective To observe the effect of intravitreal injection of mouse nerve growth factor (NGF)on interphotoreceptor retinoid-binding protein(IRBP)in the vitreous of diabetic rats at early stages.Methods Ninety-six male Sprague Dawley(SD)rats were divided into control group(group A,24 rats)and experimental group(72 rats).The rats in experimental group were induced with streptozotocin injection for diabetic retinopathy model,and then randomly divided into positive control group(group B),normal saline group(group C)and NGF group(group D),24 rats in each group.The rats in the group A and B were not intervened.The rats were received intravitreal injection with 4μl normal saline(group C)or 4 μl(0.5 μg/μ1)NGF(group D).At 2,4,6 and 8 weeks after injection,IRBP levels were detected bv enzvmelinked immunosorbent assay(ELISA);hematoxylin-eosin(HE)staining and light microscope were used to observe the morphological changes of the retina;transmission electron microscope was used to observe the retinal uhrastructure.Results At 2 weeks after injection,there was no significant difference in IRBP expression between group A,B,C and D(F=2.833,P=0.052).At 4,6,8 weeks after injection,the differences of IRBP expression between group A,B,C and D were significant(F=22.252,108.459,105.726;P=0.000).At different time points after injection,there was no significant difference in IRBP expression of group A(F=1.462,P=0.241),but there were significant differences in IRBP expression of group B.C and D(F=150.98,63.519,64.604;P=0.000).Light microscope found that the retinal structure was clear in group A and in group B,C,D at 2,4 weeks after injection;the retinal thickness were thinner in group B,C,D at 8 weeks after injection.Transmission electron microscope displayed that the structure of rod outer segments was clear in group A and in group B,C,D at 2 weeks after injection;partly unclear structure of rod outer segments and slightly enlarged gap were observed in group B,C,D at 4,8Weeks after injection.Conclusion Intravitreal injection with NGF can stabilize the IRBP expression in the vitreous of diabetic rats at early stages effectively.     

超声微泡造影剂促进重组腺相关病毒载体介导基因转染视网膜神经节细胞的体内实验

Objective To investigate the enhancing effect of ultrasound microbubbles on transfection of recombinant adeno-associated virus (rAAV) mediated green fluorecent protein (EGFP) gene into retinal ganglion cells (RGC) in vivo. Methods A total of 40 adult Sprague-Dawley (SD) rats were divided into four groups randomly (group A, B, C, D) with 10 rats in each. Group A was the normal control, in which the rats underwent intravitreal injection with 5 μl phosphate buffered solution. The rats in group B underwent intravitreal injection with 5 μl recombinant adeno-associated virus encoding EGFP gene (rAAV2-EGFP). The rats in group C underwent ultrasound irradiation on eyes right after intravitreal injection with 5μl rAAV2-EGFP; The ultrasound irradiation was performed on the rats in group D right after intravitreal injection with the mixture solution of microbubbles and rAAV2-EGFP ultrasound. After 21 days, RGC were labeled retogradely with fluogold. Seven days after labeling, the retinal flatmounts and frozen sections were made from five rats in each group. Expression of EGFP reporter gene was observed by laser scanning confocal microscope and evaluated via average optical intensity (AOD) and RGC transfection rate. Labeled RGC were counted to evaluate the adverse effects. Results Green fluorescence can be observed exactly in labeled RGC in B,C, and D groups. The AOD and transfection rate in group D was (95.02 ± 7. 25)% and (20. 10±0. 74)% , respectively; which were higher than those in group B and C (F=25. 970,25. 799;P<0.01). The difference of the number of RGC among the four groups was not significant(F= 0. 877, P>0. 05). Conclusion Under the condition of low frequency and with certain energy, ultrasound-mediated microbubble destruction can effectively and safely enhance rAAV delivery to RGC in rats.     

糖尿病视网膜病变对大鼠循环外周血内皮祖细胞数量的影响

Objective To observe the effect of diabetic retinopathy on endothelial progenitor cells (EPCs)from peripheral blood.Methods Sixty male Wistar rats were divided into control group and diabetes group.The rats in diabetes group were induced with streptozotocin(STZ)injection for diabetic retinopathy model.Flow cytometry was used to identify and count the number of EPCs from peripheral blood at 1 week.1,3 and 6 months after injection.All eyeballs were examined by hematoxylin and eosin (HE)staining,periodic acid-Schiffs(PAS)staining of trypsin-digested retinal vessels flat preparation and transmission electron microscope.EPCs count,and the relationship between DR morphological changes and EPCs count were compared and analyzed.Results The quantity of EPCs from peripheral blood at 1 week,1,3 and 6 months after STZ injection were 25±7,28±8,39±7,43±7 cells per 200 000 monocytes respectively,which decreased compared with the control group 45±4 cells per 200 000 monocytes(F=8.933,P＜0.0 1).The quantity of EPCs was gradually increased at 1 week,1,3 and 6 months after STZ injection,accompanied with responsive pathological changes of retinal structure and vessels.The thickness of retina at 1 week and 1 month after injection were reduced slightly.The number of retinal ganglion cells reduced,with the time passing by.Endothelial cells were edema,mitochondrial was swollen,capillary basement membrane was thicken,lumen was significant stenosis,lumen occlusion and retinal artery aneurysm were observed at 6 months after STZ injection.Conclusion The number of EPCs increases gradually throughout the development of DR.     

糖尿病视网膜病变对大鼠循环外周血内皮祖细胞数量的影响

Objective To observe the effect of diabetic retinopathy on endothelial progenitor cells (EPCs)from peripheral blood.Methods Sixty male Wistar rats were divided into control group and diabetes group.The rats in diabetes group were induced with streptozotocin(STZ)injection for diabetic retinopathy model.Flow cytometry was used to identify and count the number of EPCs from peripheral blood at 1 week.1,3 and 6 months after injection.All eyeballs were examined by hematoxylin and eosin (HE)staining,periodic acid-Schiffs(PAS)staining of trypsin-digested retinal vessels flat preparation and transmission electron microscope.EPCs count,and the relationship between DR morphological changes and EPCs count were compared and analyzed.Results The quantity of EPCs from peripheral blood at 1 week,1,3 and 6 months after STZ injection were 25±7,28±8,39±7,43±7 cells per 200 000 monocytes respectively,which decreased compared with the control group 45±4 cells per 200 000 monocytes(F=8.933,P＜0.0 1).The quantity of EPCs was gradually increased at 1 week,1,3 and 6 months after STZ injection,accompanied with responsive pathological changes of retinal structure and vessels.The thickness of retina at 1 week and 1 month after injection were reduced slightly.The number of retinal ganglion cells reduced,with the time passing by.Endothelial cells were edema,mitochondrial was swollen,capillary basement membrane was thicken,lumen was significant stenosis,lumen occlusion and retinal artery aneurysm were observed at 6 months after STZ injection.Conclusion The number of EPCs increases gradually throughout the development of DR.     

色素上皮衍生因子对糖尿病大鼠视网膜谷氨酸代谢的影响

Objective To observe the effect of pigment epithelium-derived factor(PEDF)on glutamate metabolism in diabetic rat retina.Methods 78 Sprague-Dawley rats were randomly divided into the model group,model control group,PEDF intervention group and intervention control group.There were some dead and euglycemia rats at the end of experiment,so only 12 rats in each group were included in the statistical analysis.The diabetic retinopathy rat model of the model,PEDF intervention and intervention control group were induced with streptozotocin injection.The rats in the model group were not intervened.The monthly-age matched normal rats of model group were in the model control group.The left eyes of rats were received intravitreal injection with 5μl(0.1μg/μl)PEDF(PEDF intervention group)or 5μlphosphate buffer solution(intervention control group).The expressions of L-glutamate/L-aspartate transporter in retina were analyzed by western blot and real time RT-PCR techniques and glutamate content in retina was analyzed by high-pressure liquid chromatography(HPLC).Cultured rat Mailer cells were divided into the control,experimental,PEDF intervention and intervention control group,GLAST expressions were detected by fluorescence immunofluorescence and real-time RT-PCR techniques.The glutamate up-take activity of Müller cells was determined by intracellular[3H] labeled D,L-glutamate concentration with scintillation counting.Results Western blot and real-time RT-PCR showed that GLAST expression decreased(real-time RT-PCR:t=8.86,P＜0.01;Western blot:t=3.42,P＜0.05).glutamate content increased(t=4.01,P＜0.05)in model group compared with the model control group:GLAST expression increased(real-time RT-PCR:t=3.56,P＜0.05;Western blot:t=3.52,P＜0.05),glutamate content decreased(y=4.36,P＜0.05)in the PEDF intervention group compared with the intervention control group.Real-time RT-PCR and fluorescence immunofluorescenee showed that high glucose down-regulate GLAST expressions in Müller cells(real-time RT-PCR:t=3.48,P＜0.05;fluorescence immunofluoreseence:t=4.72,P＜0.05)and impair glutamate up-take activity of Müller cells(t=3.81,P＜0.05).Under high glucose conditions,PEDF up-regulated GLAST expression significantly(real-time RT-PCR:t=6.82,P＜0.01;fluorescence immunofluorescence:t=3.72,P＜0.05)and ameliorated the glutamate up-take activitv of Mailer cells(t=4.14,P＜0.05).Conclusions In diabetic rats,PEDF may improve the activitv of GLAST in Müller cells,thus ameliorate retinal glutamate metabolism and inhibit death of retinal ganglion cells.     

去炎松对玻璃体切除兔眼视网膜电图和超微结构的影响

3组12只兔双眼行玻璃体切除术，分别于一眼玻璃体内注入0.5、1.0，2.0mg去炎松悬液，另一眼注入眼用平衡盐液．观察术后28天内眼底及ERGb波和28天时视网膜结构的变化．注入各剂量的去炎松在28天时均未完全吸收，0.5、1 .0mg组用药眼ERGb波在各时间点与对照眼及术前无显著差异(P>0.05)；2.0mg组用药眼较术前无显著差异(P>0.05)，较对照眼有轻度升高(t=3.3589，P>0.05)．光镜及电镜下视网膜结构无损害．表明玻璃体切除后使用有效剂量的去炎松对视网膜仍是安全的． （中华眼底病杂志，1996，12：105-107）     

正确认识和掌握高度近视黄斑病变的诊疗方法,减少高度近视患者视功能损害

以近视眼中心凹劈裂、黄斑裂孔及黄斑裂孔性视网膜脱离、近视性脉络膜新生血管为代表的高度近视黄斑病变是影响高度近视患者视功能的主要原因之一.光相干断层扫描检查有助于提升高度近视黄斑病变的认知水平.选择合适的手术时机,解除玻璃体后皮质和内界膜对视网膜的牵拉,有利于恢复视网膜的弹性,使近视眼中心凹劈裂消失.促进黄斑裂孔的闭合和视网膜复位;而抗新生血管药物与光动力疗法和(或)糖皮质激素联合应用,则是治疗近视性脉络膜新生血管的发展方向.正确认识和掌握高度近视黄斑病变的诊疗方法 和时机,采取针对性的干预措施.使其能够得到合理有效的治疗,是进一步减少高度近视患者视功能损害的关键.     

Coats病19例临床病理观察

目的：观察Coats病的临床及病理改变。 方法：复习本院眼科病理室1959—1994年经病理学证实的19例Coats病患者的临床资料和病理切片。 结果：男14例，女5例，年龄为1~18岁，10岁前发病者男性明显多于女性，随年龄增长，性别差异逐渐减少。病理改变主要是视网膜外层血管扩张充血，血管壁厚薄不均及结缔组织增生。晚期病例眼底多见视网膜脱离隆起，表面有血管改变和出血，视网膜下液中有泡沫细胞和胆固醇结晶，有的病例视网膜外层有钙化及骨化。 结论：Coats病是视网膜血管病变引起的视网膜多种病理变化，视网膜下液细胞学检查可作为本病与视网膜母细胞瘤鉴别诊断的可靠方法。 （中华眼底病杂志，1996，12：157-159）     

后巩膜加固术治疗高度近视黄斑裂孔性视网膜脱离

目的：探讨改进的后巩膜加固术对高度近视黄斑裂孔性视网膜脱离的治疗效果。 方法：1993年3月至1995年11月对住院的20例高度近视黄斑裂孔性视网膜脱离患者的20只眼，采用后巩膜加固术治疗，有关临床资料进行回顾分析。 结果：眼轴可对比的14只眼，术前与出院时的均值为28.22mm与26.87mm，17只眼视网膜复位，获得0.02一0.2的矫正视力，3只眼失败，增殖性玻璃体视网膜病变(proliferative vitreous retinopathy，PVR)加重。 结论：后巩膜加固术可有效治疗无严重PVR的高度近视黄斑裂孔所致的视网膜脱离，不必采用视网膜粘连法以尽量保存残留的中心视力。 （中华眼底病杂志，1996，12：214-216）     

兔视网膜中蛋白激酶C的定位研究

目的：证实兔视网膜中有α、β和γ三种蛋白激酶C(protein kinase C，PKC)亚型的分布。方法；采用免疫组织化学技术，以抗PKC同功酶I(α)、Ⅱ(β)、Ⅲ(γ)单克隆抗体对兔视网膜进行PKC同功酶定位研究。 结果：兔视网膜中有PKC-α、PKC-β和PKC-γ的阳性免疫反应。PKC-α免疫反应主要呈现在内核层的双极细胞；PKC-β免疫反应主要呈现在神经节细胞层的神经节细胞；PKC-γ则在神经节细胞层、内网状层以及光感受器的外节呈弥漫性弱阳性染色。 结论：作为中枢神经系统的一部分，视网膜可以同时存在PKC-α、PKC-β和PKC-γ三种亚型。 （中华眼底病杂志，1996，12：242-244）     

病理性近视黄斑劈裂光相干断层扫描图像分型及其对临床的指导意义

病理性近视黄斑劈裂光相干断层扫描(OCT)的类组织学分型可以分为单纯外层劈裂、外层和中层劈裂、外层和内层劈裂、多层劈裂4种情况.以彻底清除玻璃体后皮质及后部血管弓内的内界膜为重点的玻璃体视网手术是治疗病理性近视黄斑劈裂的主要选择.其中,单纯外层劈裂合并中心凹脱离者,手术后视功能改善较明显;而多层劈裂者视功能改善有限或不改善.合理剥除后皮质与内界膜起始点,应在未发生内层劈裂处.正确认识和了解病理性近视黄斑劈裂的OCT分型.对于选择玻璃体视网膜手术方式和判断治疗预后有积极意义.     

脉络膜黑色素瘤组织病理学分析

目的 观察脉络膜黑色素瘤的组织病理学特征。 方法 回顾分析64例病理确诊的脉络膜黑色素瘤病理资料。按照美国眼黑色素瘤多中心研究组的测量方法和WHO的分类标准，测量和观察大体标本中的肿瘤大小和光学显微镜下的肿瘤细胞学类型；以肿瘤前缘累及部位对肿瘤所处位置进行分类；以肿瘤细胞向外浸润程度对肿瘤蔓延程度进行分级。 结果 64例脉络膜黑色素瘤中，大肿瘤25例，占39.1%；中等大小肿瘤31例，占48.4%；小肿瘤8例，占12.5%。梭形细胞型42例，占65.6%,其中，梭形细胞A型15例,占23.4%，梭形细胞B型27例,占42.3%;上皮样细胞型7例,占10.9%;混合型10例,占15.6%;其它型5例,占7.8%。肿瘤细胞未累及巩膜者25例,占39.1%;累及巩膜但限于巩膜层内者22例,占34.4%;穿透巩膜全层达眼球表面者12例,占18.7%;眶内浸润者5例,占7.8%。 结论 脉络膜黑色素瘤组织病理学特征变化多样，临床上以梭形细胞型最常见，易伴巩膜浸润。 (中华眼底病杂志, 2006, 22: 161-165)     

前后段联合手术及硅油填充治疗高度近视黄斑孔视网膜脱离的疗效观察

目的 观察前后段联合手术及硅油充填治疗高度近视黄斑孔视网膜脱离临床疗效.方法 回顾分析前后段联合手术及硅油充填治疗高度近视黄斑孔视网膜脱离患者48例48只眼的临床资料.患者均有高度近视史,视网膜脱离以后极部为主.裂隙灯前置镜和(或)光相干断层扫描(OCT)检查均发现黄斑裂孔.均行白内障超声乳化或抽吸联合玻璃体切割硅油充填,41例行内界膜(ILM)剥离,23例植入人工晶状体(10L).硅油取出的时间距第一次手术时间为3.5～48.0个月.取硅油前均行OCT检查.取硅油后随访观察均1年以上.结果 除5例外,其他患者手术后1周,前置镜检查均不能看到黄斑孔边缘;视力均有不同程度的提高.48例患者全部已取硅油.取硅油前OCT检查,黄斑孔愈合呈U型8例,V型为6例,W型为23例;未闭合11例.未闭合的11例经取硅油与膨胀气体充填后全部复位,其中,U型2例,W型9例.32例W型愈合者中2例患者在取油后13、38个月后出现视网膜脱离复发.最终黄斑裂孔U型和V型愈合者16例,占33.3%;W型愈合者32例,占66.7%.视网膜复位率为100.0%.结论前后段联合手术及硅油充填是治疗高度近视黄斑孔视网膜脱离的有效方法 ;OCT检查是确定黄斑孔是否封闭的客观标准.     

病理性近视黄斑裂孔患者黄斑区视网膜前膜的组织学观察

目的 观察病理性近视内界膜表面结构的组织学变化与黄斑裂孔发生发展的关系.方法 同顾分析行玻璃体切割手术的病理性近视黄斑裂孔患者34例34只眼的临床资料.患眼屈光度均超过-6.00 D,眼轴26.00～33.12 mm,平均眼轴长度27.74 mm.5只眼为黄斑裂孔无视网膜脱离(黄斑裂孔组),29只眼为黄斑裂孔合并后极部视网膜浅脱离(视网膜脱离组).对入选患眼行睫状体平坦部三切口的玻璃体切割手术,手术中观察Weiss环以判断玻璃体后脱离程度,获取34只眼的视网膜前膜及19只眼的内界膜组织标本,行苏木精-伊红(HE)及醋酸铀-枸橼酸铅双染色,采用光学及透射电子显微镜观察.结果 玻璃体切割手术中,5只眼出现Weiss环,24只眼的视网膜表面有多层玻璃体组织残留.光学显微镜观察发现,内界膜表面的视网膜前膜主要由玻璃体胶原纤维.星形胶质细胞及细胞外基质组成.透射电子显微镜观察发现,19只眼的内界膜标本中,5只眼可她内界膜一玻璃体胶原纤维-细胞的"三明治"样结构,1只眼可见内界膜损伤、表面组织牵引和星形胶质细胞移行.结论病理性近视玻璃体后界面劈裂、内界膜表面组织结构的变化是黄斑裂孔发生发展直至视网膜脱离的重要原因.     

视网膜内界膜剥离手术治疗高度近视黄斑裂孔视网膜脱离的疗效观察

目的 观察视网膜内界膜剥离治疗高度近视黄斑裂孔视网膜脱离的疗效. 方法 回顾分析25例25只眼高度近视黄斑裂孔伴视网膜脱离患者的临床资料.根据治疗方法 分为2组,A组为单纯玻璃体切割手术,13例13只眼;B组为玻璃体切割手术加吲哚青绿染色内界膜剥离,12例12只眼.所有患者行惰性气体填充,手术后保持面朝下体位7～15 d.观察最佳矫正分辨角对数(LogMAR)视力,检查眼底,光相干断层扫描(OCT)、B型超声检查视网膜复位及黄斑裂孔闭合情况,比较两组间疗效差异.手术后随访6～18个月,平均随访时间10个月. 结果 A组13只眼中,7只眼手术后视网膜复位,占53.8%;B组12只眼中,11只眼手术后视网膜复位,占91.7%.B组视网膜复位率明显优于A组(X2=4.427,P=0.046);25只眼中,手术后黄斑裂孔闭合者17只眼,占68.0%.其中,A组13只眼中,6只眼黄斑裂孔闭合,占A组患者的46.2%;B组12只眼中,11只眼黄斑裂孔闭合,占B组患者的91.7%.两组患者手术后黄斑裂孔闭合率比较,差异有统计学意义(X2=5.940,P=0.020).A组手术后最佳矫正LogMAR视力提高平均0.32,与手术前比较,差异有统计学意义(Z=-2.045,P=0.041),B组手术后最佳矫正LogMAR视力提高平均0.53,与手术前比较,差异有统计学意义(Z=-2.481,P=0.012).两组间手术后视力差异无统计学意义(U=51.5,P=0.16). 结论玻璃体切割联合视网膜内界膜剥离手术可能通过完全解除玻璃体黄斑牵引、增加视网膜顺应性而提高高度近视黄斑裂孔视网膜脱离的治疗效果.显著增加视网膜复位率及黄斑裂孔闭合率.