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
The antibiotics, metronidazole and ciprofloxacin, are the first-line treatment for pouchitis. Patients who do not respond to antibiotics or conventional medications represent a major challenge to therapy. In this repo...The antibiotics, metronidazole and ciprofloxacin, are the first-line treatment for pouchitis. Patients who do not respond to antibiotics or conventional medications represent a major challenge to therapy. In this report, we have described a successful treatment of severe refractory pouchitis with a novel agent, rebamipide, known to promote epithelial cell regeneration and angiogenesis. A 27-year-old male with ileo-anal pouch surgery presented with worsening anal pain, diarrhea, and abdominal pain. The patient was diagnosed to have pouchitis and was given metronidazole together with betamethasone enema （3.95 rag/dose）. However, despite this intensive therapy, the patient did not improve. On endoscopy, ulceration and inflammation were seen in the ileal pouch together with contact bleeding and mucous discharge. The patient was treated with rebamipide enema （150 rag/close） twice a clay for 8 wk without additional drug therapy. Two weeks after the rebamipide therapy, stool frequency started to decrease and fecal hemoglobin became negative at the 4^th wk. At the end of the therapy, endoscopy revealed that ulcers in the ileal pouch had healed with no obvious inflammation. The effect of rebamipide enema was dramatic and was maintained throughout the ll-mo follow-up. The patient continued to be in remission. No adverse effects were observed during the treatment or the follow-up period. The sustained response seen in this case with severe and refractory pouchitis indicates that agents, which promote epithelial cell growth, angiogenesis and mucosal tissue regeneration, are potential therapeutic agents for the treatment of refractory colorectal lesions.展开更多
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。
文摘The antibiotics, metronidazole and ciprofloxacin, are the first-line treatment for pouchitis. Patients who do not respond to antibiotics or conventional medications represent a major challenge to therapy. In this report, we have described a successful treatment of severe refractory pouchitis with a novel agent, rebamipide, known to promote epithelial cell regeneration and angiogenesis. A 27-year-old male with ileo-anal pouch surgery presented with worsening anal pain, diarrhea, and abdominal pain. The patient was diagnosed to have pouchitis and was given metronidazole together with betamethasone enema （3.95 rag/dose）. However, despite this intensive therapy, the patient did not improve. On endoscopy, ulceration and inflammation were seen in the ileal pouch together with contact bleeding and mucous discharge. The patient was treated with rebamipide enema （150 rag/close） twice a clay for 8 wk without additional drug therapy. Two weeks after the rebamipide therapy, stool frequency started to decrease and fecal hemoglobin became negative at the 4^th wk. At the end of the therapy, endoscopy revealed that ulcers in the ileal pouch had healed with no obvious inflammation. The effect of rebamipide enema was dramatic and was maintained throughout the ll-mo follow-up. The patient continued to be in remission. No adverse effects were observed during the treatment or the follow-up period. The sustained response seen in this case with severe and refractory pouchitis indicates that agents, which promote epithelial cell growth, angiogenesis and mucosal tissue regeneration, are potential therapeutic agents for the treatment of refractory colorectal lesions.
