目的探讨CRISPR/CAS9靶向敲除Oct3/4基因后化疗药物索拉菲尼(Sorafenib)对肝癌细胞的作用影响。方法通过实时荧光定量PCR和免疫印迹检测人肝癌细胞Li-7、HepG2、Huh7、BEL-7405和人肝正常细胞LX-2中Oct3/4的表达水平。通过CRISPR设计工...目的探讨CRISPR/CAS9靶向敲除Oct3/4基因后化疗药物索拉菲尼(Sorafenib)对肝癌细胞的作用影响。方法通过实时荧光定量PCR和免疫印迹检测人肝癌细胞Li-7、HepG2、Huh7、BEL-7405和人肝正常细胞LX-2中Oct3/4的表达水平。通过CRISPR设计工具(http://crispr.mit.edu/)设计靶向Oct3/4的gRNA,并通过CRISPR/CAS9技术敲除肝癌细胞HepG2的Oct3/4,评估Oct3/4缺失情况下,化疗药物Sorafenib对肝癌细胞HepG2的凋亡水平和DNA损伤水平的影响。结果Oct3/4在人肝癌细胞Li-7、HepG2、Huh7、BEL-7405中的mRNA表达水平显著高于人肝正常细胞LX-2(P<0.05);同时,Oct3/4在人肝癌细胞Li-7、HepG2、Huh7、BEL-7405中的蛋白表达水平也显著高于人肝正常细胞LX-2。使用CRISPR/CAS9对HepG2细胞进行基因编辑后,挑取嘌呤霉素筛选出的2个单克隆细胞株进行Oct3/4表达水平的检测,1#号单克隆细胞株Oct3/4表达水平低于野生型WT,2#号单克隆细胞株没有表达Oct3/4。测序后发现1#号基因组上的Oct3/4第1个外显子只有1条染色体缺失了2个碱基,另一条染色体并未缺失;2#号基因组上的Oct3/4第1个外显子2条染色体均存在不同程度的缺失,一条缺失2个碱基,另一条缺失4个碱基。使用Sorafenib处理野生型HepG2细胞和2#Oct3/4 KO HepG2细胞后,Oct3/4 KO细胞株的凋亡水平显著上升(P<0.05),DNA损伤水平显著上升(P<0.05)。结论Oct3/4基因的敲除能够显著提高化疗药物Sorafenib促肝癌细胞凋亡和DNA损伤的作用。展开更多
Aims. The authors conducted a study to compare retinal thickness in patients with choroidal neovascularization (CNV) using optical coherence tomography (OCT) and retinal thickness analyser (RTA). Methods. In all, 11 e...Aims. The authors conducted a study to compare retinal thickness in patients with choroidal neovascularization (CNV) using optical coherence tomography (OCT) and retinal thickness analyser (RTA). Methods. In all, 11 eyes from 11 patients with subfoveal choroidal neovascularization were examined with OCT and RTA. Results. Three patients could not be explored by RTA due to lack of fixation and high myopia. The mean foveal thickness was 289.9± 92.1 μ m with OCT and 207.7± 60.8 μ m with RTA. The mean perifoveal thickness was 293.8± 46.3 μ m and 200.8± 61.3 μ m, respectively. The maximal perifoveal thickness measured by OCT was 335± 104.0 μ m and 316± 86.8 μ m by RTA. Conclusion. OCT and RTA are able to detect increases in retinal thickness due to the presence of CNV and of fluid extravasation. They can be used in measuring retinal thickness in patients with CNV, although measures are not comparable between both systems.展开更多
Purpose To report the morphologic data of adult-onset foveomacular vitelliform dystrophy(AFVD)provided by thirdgeneration optical coherence tomography(OCT 3).Design Observational case report.Methods An 85-year-old wom...Purpose To report the morphologic data of adult-onset foveomacular vitelliform dystrophy(AFVD)provided by thirdgeneration optical coherence tomography(OCT 3).Design Observational case report.Methods An 85-year-old woman presenting with AFVD underwen t fundus biomi-croscopy,fluorescein angiography,and OCT examination.Results Fundus examination disclosed a round,elevated yellowish lesion,centered by a pigm ented spot,in the macula of the left eye.Examination with OCT disclosed an area of hyperreflectivity located b etween the retinal pigment epithelium layer and the photorecep tor layer,compatible in size with the yellowish elevated lesion.The hyperreflective line corresponding to the photoreceptor layer was elevated by the material and separated from th e retinal pigment ep-ithelium layer.Conclusions In AFVD,examination with OCT 3demonstrates elevation of the p hotoreceptor layer by the material and supports the previo us hypothesis that the material is located between the photoreceptor and the reti-nal pigment epithelium layer.展开更多
文摘目的:探讨海德堡激光眼底扫描仪(HRT-Ⅱ)和光学相干断层扫描仪(OCT3)检测视网膜神经纤维层厚度(retinal nerve fiber layer,RNFL)在青光眼早期诊断中的价值。方法:可疑开角型青光眼(suspected open angle glaucoma,SOAG)48例、原发性开角型青光眼(primary open angle glaucoma,POAG)55例与年龄相匹配的正常人43例。采用Oculus-Easyfield视野计flied视野计、HRT-Ⅱ和OCT3分别进行视野、视盘形态和视网膜神经纤维层厚度的检测。选用HRT-Ⅱ所测视盘参数中的平均mRNFL、截面面积RNFLA和OCT3所测的mRNFL做比较分析。测的平均视网膜神经纤维层厚度(RNFL)做分析比较。采用方差分析,对HRT-Ⅱ和OCT3检测3组间的参数进行两两比较;对HRT-Ⅱ视盘参数中的mRNFL,RNFLA和OCT3所测的mRNFL厚度及各自与视野平均缺损值(meandefect,MD)做相关分析。结果:HRT-Ⅱ视盘参数中的mRNFL,RNFLA及OCT3所测的mRNFL均存在显性差异(P<0.05);在POAG组,HRT-Ⅱ的mRNFL,RNFLA及OCT3所测的mRNFL显著性相关(P<0.05),且与视野平均缺损值(MD)有不同程度的相关。结论:HRT-Ⅱ和OCT3均能够区分青光眼的不同状态。HRT-Ⅱ所测mRNFL,RNFLA与OCT3所测的mRNFL有较好的相关性且均与视野平均缺损值相关。
文摘目的探讨CRISPR/CAS9靶向敲除Oct3/4基因后化疗药物索拉菲尼(Sorafenib)对肝癌细胞的作用影响。方法通过实时荧光定量PCR和免疫印迹检测人肝癌细胞Li-7、HepG2、Huh7、BEL-7405和人肝正常细胞LX-2中Oct3/4的表达水平。通过CRISPR设计工具(http://crispr.mit.edu/)设计靶向Oct3/4的gRNA,并通过CRISPR/CAS9技术敲除肝癌细胞HepG2的Oct3/4,评估Oct3/4缺失情况下,化疗药物Sorafenib对肝癌细胞HepG2的凋亡水平和DNA损伤水平的影响。结果Oct3/4在人肝癌细胞Li-7、HepG2、Huh7、BEL-7405中的mRNA表达水平显著高于人肝正常细胞LX-2(P<0.05);同时,Oct3/4在人肝癌细胞Li-7、HepG2、Huh7、BEL-7405中的蛋白表达水平也显著高于人肝正常细胞LX-2。使用CRISPR/CAS9对HepG2细胞进行基因编辑后,挑取嘌呤霉素筛选出的2个单克隆细胞株进行Oct3/4表达水平的检测,1#号单克隆细胞株Oct3/4表达水平低于野生型WT,2#号单克隆细胞株没有表达Oct3/4。测序后发现1#号基因组上的Oct3/4第1个外显子只有1条染色体缺失了2个碱基,另一条染色体并未缺失;2#号基因组上的Oct3/4第1个外显子2条染色体均存在不同程度的缺失,一条缺失2个碱基,另一条缺失4个碱基。使用Sorafenib处理野生型HepG2细胞和2#Oct3/4 KO HepG2细胞后,Oct3/4 KO细胞株的凋亡水平显著上升(P<0.05),DNA损伤水平显著上升(P<0.05)。结论Oct3/4基因的敲除能够显著提高化疗药物Sorafenib促肝癌细胞凋亡和DNA损伤的作用。
文摘Aims. The authors conducted a study to compare retinal thickness in patients with choroidal neovascularization (CNV) using optical coherence tomography (OCT) and retinal thickness analyser (RTA). Methods. In all, 11 eyes from 11 patients with subfoveal choroidal neovascularization were examined with OCT and RTA. Results. Three patients could not be explored by RTA due to lack of fixation and high myopia. The mean foveal thickness was 289.9± 92.1 μ m with OCT and 207.7± 60.8 μ m with RTA. The mean perifoveal thickness was 293.8± 46.3 μ m and 200.8± 61.3 μ m, respectively. The maximal perifoveal thickness measured by OCT was 335± 104.0 μ m and 316± 86.8 μ m by RTA. Conclusion. OCT and RTA are able to detect increases in retinal thickness due to the presence of CNV and of fluid extravasation. They can be used in measuring retinal thickness in patients with CNV, although measures are not comparable between both systems.
文摘Purpose To report the morphologic data of adult-onset foveomacular vitelliform dystrophy(AFVD)provided by thirdgeneration optical coherence tomography(OCT 3).Design Observational case report.Methods An 85-year-old woman presenting with AFVD underwen t fundus biomi-croscopy,fluorescein angiography,and OCT examination.Results Fundus examination disclosed a round,elevated yellowish lesion,centered by a pigm ented spot,in the macula of the left eye.Examination with OCT disclosed an area of hyperreflectivity located b etween the retinal pigment epithelium layer and the photorecep tor layer,compatible in size with the yellowish elevated lesion.The hyperreflective line corresponding to the photoreceptor layer was elevated by the material and separated from th e retinal pigment ep-ithelium layer.Conclusions In AFVD,examination with OCT 3demonstrates elevation of the p hotoreceptor layer by the material and supports the previo us hypothesis that the material is located between the photoreceptor and the reti-nal pigment epithelium layer.
