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基于生物信息学分析揭示ceRNA网络调控锰致神经毒性的标志物
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作者 张诗玄 李婕 +4 位作者 武璐丽 马俊香 陈丽 肖忠新 牛丕业 《环境卫生学杂志》 2022年第4期247-253,共7页
目的筛选锰(manganese,Mn)染毒人神经母细胞瘤(SH-SY5Y)细胞后差异表达RNAs,分析内源性竞争RNA(competing endogenous RNA,ceRNA)共表达网络并鉴定出与帕金森病(Parkinson’s disease,PD)患者大脑皮质中共表达的关键RNAs,揭示Mn致神经... 目的筛选锰(manganese,Mn)染毒人神经母细胞瘤(SH-SY5Y)细胞后差异表达RNAs,分析内源性竞争RNA(competing endogenous RNA,ceRNA)共表达网络并鉴定出与帕金森病(Parkinson’s disease,PD)患者大脑皮质中共表达的关键RNAs,揭示Mn致神经毒性与PD的共同分子机制和潜在的生物标志物。方法利用Agilent人表达谱芯片对Mn暴露(500μmol/L MnCl_(2),6h)SH-SY5Y细胞进行mRNAs、miRNAs和lncRNAs检测;运用R软件分析差异表达(differential expression,DE)基因。通过starBase和miRTarBase数据库鉴定miRNA-lncRNA和miRNA-mRNA相互作用关系,用Cytoscape软件构建ceRNA调控网络,对比分析GEO数据库GSE72962检测的PD患者额叶皮质miRNAs表达数据;对DE mRNAs进行GO功能富集和KEGG通路分析。结果Mn染毒SH-SY5Y细胞后差异表达分析筛选出DE lncRNAs 177个,DE miRNAs 112个和DE mRNAs 6353个。根据共表达分析结果和ceRNA调控机制,最终纳入lncRNAs 20个,miRNAs 14个和mRNAs 27个,共65对相互作用关系构建lncRNA-miRNA-mRNA调控网络。其中miR-124-3p和miR-371a-5p的表达在Mn染毒神经细胞和PD患者额叶皮质中均有显著性变化。GO和KEGG富集分析提示,ceRNA主要富集在神经系统发育、凋亡信号调节、氧化应激反应、自噬、细胞衰老等通路。结论ceRNA调控网络在Mn致神经毒性作用中发挥着重要作用,miR-124-3p和miR-371a-5p可能会成为锰中毒和PD诊断的生物标志物。 展开更多
关键词 神经毒性 帕金森病(PD) 竞争性内源RNA(ceRNA)
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激光共聚焦显微镜对样品检测中的温度变化及不同固定方法的研究 被引量:2
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作者 肖忠新 赵媛媛 《中国医学装备》 2018年第12期161-163,共3页
目的:研究环境温度的变化和样本固定的方法对于激光共聚焦显微镜检测样品的影响。方法:选取1个铃兰样片,对在冷冻保存和常温下的样片进行荧光强度检测。将样片保存于冰箱-20℃冷冻12 h,取出恢复至室温后利用激光共聚焦显微镜观测样片荧... 目的:研究环境温度的变化和样本固定的方法对于激光共聚焦显微镜检测样品的影响。方法:选取1个铃兰样片,对在冷冻保存和常温下的样片进行荧光强度检测。将样片保存于冰箱-20℃冷冻12 h,取出恢复至室温后利用激光共聚焦显微镜观测样片荧光强度变化;采集室温下铃兰样片图,观测样片荧光强度;样片观测时间间隔5 min,相同视野下采集图像,共采集30 min。依据处理方法的不同将其分为A组和B组,A组为甲醇处理组(3个皿);B组为甲醛处理组(3个皿),分别采用4%多聚甲醛缓冲液、预冷的甲醇两种固定剂,固定人正常肺上皮细胞,同时两组各再加渗透试剂TritonX-100处理细胞,用4',6-二脒基-2-苯基吲哚(DAPI)复染液对肺上皮细胞核染色;在相同显微镜的物镜和样品荧光激发强度及图像采集参数条件下,使用激光共聚焦显微镜观测、采集图像。结果:在冰箱-20℃冷冻保存的样片的荧光强度呈逐步上升趋势,室温下样片的荧光强度无明显改变。两组固定剂再加渗透试剂TritonX-100处理,细胞核染色效果更好。结论:环境温度的变化会影响样本荧光强度,普通固定剂固定细胞之后再加渗透试剂处理有利于细胞核染色。 展开更多
关键词 温度的变化 固定的方法 激光共聚焦显微镜 荧光强度
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Mutagenicity of Urine From Individuals Exposed to LPG Combustion Products
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作者 YIN XUE-JUN LIU JUN-ZHUO +3 位作者 KONG XIANG-HUAN CHU JIN-HUA WANG HONG xiao zhong-xin 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 1998年第3期251-257,共7页
The mutngenicity of urine from individua1s exposed to the combustion products of liquefied petroleum gas (LPG) was detected with Salmonella typhimurium TA98 and its newly developed derivatives YG1021 (nitroreductase o... The mutngenicity of urine from individua1s exposed to the combustion products of liquefied petroleum gas (LPG) was detected with Salmonella typhimurium TA98 and its newly developed derivatives YG1021 (nitroreductase overproducing) and YG1024 (O-acetyltransferase overproducing). The detection showed significantly increased mutagenicity for the two YG strains and increased positive rates for all three stralns in the presence of both rat liver S9 and β-glucuronidase. Further analysis demonstrated that urine samples taken from smoking and non-smoking exposed individuals exhibited significantly higher mutagenic potency (revertants/10 μl urine concentrate) than their corresponding controls. These results indicate that the increased urine mutagenicity is caused by the exposure to LPG combustion products or smoking. The mutagenic potency of urine samples of all exposed individuals tested with YG1024 was found to be about 7 times higher than with TA98. The difference in mutagenic potency was smaller for the sarne samples when comparison was made between YG1021 and TA98. This suggests that the mutagenic compounds present in the urine samples contaln mainly arotnatic compounds as glucuronide conugates. our results demonstrate that YG1024 is more sensitive than TA98 in detecting the mutagenicity of these samples. In addition, no significant difference in the mutagenic poency between the' pure' exposed (non-srnokers') and the ' pure' smokers' (unexpeed) samples was found in all three tester strains. This might mean that the exopure extent Of mutagens/carcinogens in LPG combustion products for exmped individuals roughly corresponds to the smoking level of smokers who smoke 20-40 cigarettes per day. Furthermore, the results also suggest that sguergism might exist in the mutagenic effects of exposure to LPG combustion products and cigarette smoking. 展开更多
关键词 NSE Mutagenicity of Urine From Individuals Exposed to LPG Combustion Products LPG
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