Congenital cataract is a crystallin severe blinding disease and genetic factors in disease development are important. Crystallin growth is under a combination of genes and their products in time and space to complete ...Congenital cataract is a crystallin severe blinding disease and genetic factors in disease development are important. Crystallin growth is under a combination of genes and their products in time and space to complete the coordination role of the guidance. Congenital cataract-related genes, included crystallin protein gene (CRYAA, CRYAB, CRYBA1/A3, CRYBA4, CRYBB1, CRYBB2, CRYBB3, CRYGC, CRYGD, CRYGS), gap junction channel protein gene (GJA1, GJA3, GJA8), membrane protein gene (GJA3, GJA8, MIP, LIM2), cytoskeletal protein gene (BF-SP2), transcription factor genes (HSF4, MAF, PITX3, PAX6), ferritin light chain gene (FTL), fibroblast growth factor (FGF) and so on. Currently, there are about 39 genetic loci isolated to which primary cataracts have been mapped, although the number is constantly increasing and depends to some extent on definition. We summarized the recent advances on epidemiology and genetic locations of congenital cataract in this review.展开更多
目的研究Ⅱ型重组腺相关病毒（recombinant adeno-associated virus 2，rAAV2）载体介导的增强型绿色荧光蛋白基因（enhanced green fluorescent protein，ECFP）对体外培养兔晶状体上皮细胞的转染和表达情况，为rAAV携带目的基因防治...目的研究Ⅱ型重组腺相关病毒（recombinant adeno-associated virus 2，rAAV2）载体介导的增强型绿色荧光蛋白基因（enhanced green fluorescent protein，ECFP）对体外培养兔晶状体上皮细胞的转染和表达情况，为rAAV携带目的基因防治后囊膜混浊提供理论依据。方法组织块培养法体外培养兔晶状体上皮细胞。rAAV2-EGFP，按感染复数（multiplicity of infection，MOI）为10^3、10^4、10^5、4×10^5、10^6转染传2代细胞，转染后第1天、第3天、第5天、第7天、第9天、第11天在倒置荧光显微镜下观察晶状体上皮细胞中EGFP表达的阳性情况，记录200个细胞中EGFP阳性表达所占的百分比。当EGFP表达稳定后用激光共聚焦显微镜照相。用倒置显微镜观察rAAV转染对细胞生长和形态的影响。结果当MOI=10^3、10^4时，转染后第72小时晶状体上皮细胞中EGFP开始表达，当MOI=10^5、4×10^5、10^6时，转染后第24小时便可见EGFP阳性表达。随着MOI值的增大及时间的延长，EGFP表达效率逐渐增高，转染后第8-第9天达到高峰并维持，此时，MOI=10^3、10^4、10^5、4×10^5、10^6的转导效率分别为16％、41％、55％、86％、94％。对照组和转染组细胞的生长和形态特征无明显改变。结论腺相关病毒载体可以携带报告基因稳定转染晶状体上皮细胞，并且转染效率高，因而腺相关病毒携带目的基因防治后囊膜混浊是可行的。展开更多
文摘Congenital cataract is a crystallin severe blinding disease and genetic factors in disease development are important. Crystallin growth is under a combination of genes and their products in time and space to complete the coordination role of the guidance. Congenital cataract-related genes, included crystallin protein gene (CRYAA, CRYAB, CRYBA1/A3, CRYBA4, CRYBB1, CRYBB2, CRYBB3, CRYGC, CRYGD, CRYGS), gap junction channel protein gene (GJA1, GJA3, GJA8), membrane protein gene (GJA3, GJA8, MIP, LIM2), cytoskeletal protein gene (BF-SP2), transcription factor genes (HSF4, MAF, PITX3, PAX6), ferritin light chain gene (FTL), fibroblast growth factor (FGF) and so on. Currently, there are about 39 genetic loci isolated to which primary cataracts have been mapped, although the number is constantly increasing and depends to some extent on definition. We summarized the recent advances on epidemiology and genetic locations of congenital cataract in this review.
文摘目的研究Ⅱ型重组腺相关病毒（recombinant adeno-associated virus 2，rAAV2）载体介导的增强型绿色荧光蛋白基因（enhanced green fluorescent protein，ECFP）对体外培养兔晶状体上皮细胞的转染和表达情况，为rAAV携带目的基因防治后囊膜混浊提供理论依据。方法组织块培养法体外培养兔晶状体上皮细胞。rAAV2-EGFP，按感染复数（multiplicity of infection，MOI）为10^3、10^4、10^5、4×10^5、10^6转染传2代细胞，转染后第1天、第3天、第5天、第7天、第9天、第11天在倒置荧光显微镜下观察晶状体上皮细胞中EGFP表达的阳性情况，记录200个细胞中EGFP阳性表达所占的百分比。当EGFP表达稳定后用激光共聚焦显微镜照相。用倒置显微镜观察rAAV转染对细胞生长和形态的影响。结果当MOI=10^3、10^4时，转染后第72小时晶状体上皮细胞中EGFP开始表达，当MOI=10^5、4×10^5、10^6时，转染后第24小时便可见EGFP阳性表达。随着MOI值的增大及时间的延长，EGFP表达效率逐渐增高，转染后第8-第9天达到高峰并维持，此时，MOI=10^3、10^4、10^5、4×10^5、10^6的转导效率分别为16％、41％、55％、86％、94％。对照组和转染组细胞的生长和形态特征无明显改变。结论腺相关病毒载体可以携带报告基因稳定转染晶状体上皮细胞，并且转染效率高，因而腺相关病毒携带目的基因防治后囊膜混浊是可行的。
