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8周负重爬梯抗阻训练对小鼠右心室心肌特定miRNAs的影响
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作者 宋彦霖 张海鹏 +3 位作者 徐琳 白荣雁 徐盛嘉 马继政 《湖北体育科技》 2023年第2期106-111,共6页
目的 在剧烈运动期间,右心室会承受更多的后负荷和室壁应力,本研究评定8周抗阻训练对小鼠右心室心肌特定mi RNAs的表达产生的影响,是否参与调控右心室心肌肥大。方法 12只C57BL/6J雄性小鼠,依据体重随机分为对照组(CON组)和抗阻训练组(... 目的 在剧烈运动期间,右心室会承受更多的后负荷和室壁应力,本研究评定8周抗阻训练对小鼠右心室心肌特定mi RNAs的表达产生的影响,是否参与调控右心室心肌肥大。方法 12只C57BL/6J雄性小鼠,依据体重随机分为对照组(CON组)和抗阻训练组(LRT组),每组6只。两组在预适应、抓力测试和最大自愿承载力测试后,LRT组进行8周的尾部负重爬梯训练,CON组正常饲养不进行爬梯训练。记录两组小鼠1到9周的体重、第9周的体脂率以及第1、5、9周的抓力和最大自愿承载力测试成绩,在所有测试结束48h后,取小鼠右心室心肌,利用实时荧光定量PCR检测心肌肌组织特异的mi RNAs及其他mi RNAs的表达变化情况。使用免疫印迹检测PI3K、Akt和m TOR蛋白表达量变化情况。结果 训练8周后,LRT组小鼠心脏湿重显著升高(p<0.01);抓力和最大自愿承载力测试成绩显著上升(p<0.05;p<0.01);与CON组相比,LRT组右心室mi R-208a、mi R-21、mi R-222、mi R-486表达显著上升(p<0.01;p<0.01;p<0.05;p<0.001),分别升高了0.9倍、0.8倍、0.5倍和0.7倍。其中,mi R-208a和mi R-222与心脏湿重呈正相关(r=0.8391,p<0.05;r=0.8147,p<0.05)。mi R-208b、mi R-499、mi R-1、mi R-17-3p、mi R-133a和mi R-133b在抗阻训练后的右心室心肌中没有显著变化(p>0.05)。抗阻训练不影响这些mi RNAs的表达。LRT组小鼠右心室心肌中PI3K、Akt和m TOR蛋白表达量显著升高(p<0.01;p<0.05;p<0.05)。结论8周尾部负重抗阻训练能够诱导小鼠心脏右心室肥大且激活PI3K/Akt/mTOR信号通路;mi R-208a和mi R-222可能参与调控抗阻运动引起的心脏右心室肥大。 展开更多
关键词 mi rnas 右心室心肌 抗阻训练 小鼠实验
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miR-30c promotes Schwann cell remyelination following peripheral nerve injury 被引量:10
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作者 Sheng Yi Qi-hui Wang +4 位作者 Li-li Zhao Jing Qin Ya-xian Wang Bin Yu Song-lin Zhou 《Neural Regeneration Research》 SCIE CAS CSCD 2017年第10期1708-1715,共8页
Differential expression of mi RNAs occurs in injured proximal nerve stumps and includes mi RNAs that are firstly down-regulated and then gradually up-regulated following nerve injury.These mi RNAs might be related to ... Differential expression of mi RNAs occurs in injured proximal nerve stumps and includes mi RNAs that are firstly down-regulated and then gradually up-regulated following nerve injury.These mi RNAs might be related to a Schwann cell phenotypic switch.mi R-30 c,as a member of this group,was further investigated in the current study.Sprague-Dawley rats underwent sciatic nerve transection and proximal nerve stumps were collected at 1,4,7,14,21,and 28 days post injury for analysis.Following sciatic nerve injury,mi R-30 c was down-regulated,reaching a minimum on day 4,and was then upregulated to normal levels.Schwann cells were isolated from neonatal rat sciatic nerve stumps,then transfected with mi R-30 c agomir and co-cultured in vitro with dorsal root ganglia.The enhanced expression of mi R-30 c robustly increased the amount of myelin-associated protein in the co-cultured dorsal root ganglia and Schwann cells.We then modeled sciatic nerve crush injury in vivo in Sprague-Dawley rats and tested the effect of perineural injection of mi R-30 c agomir on myelin sheath regeneration.Fourteen days after surgery,sciatic nerve stumps were harvested and subjected to immunohistochemistry,western blot analysis,and transmission electron microscopy.The direct injection of mi R-30 c stimulated the formation of myelin sheath,thus contributing to peripheral nerve regeneration.Overall,our findings indicate that mi R-30 c can promote Schwann cell myelination following peripheral nerve injury.The functional study of mi R-30 c will benefit the discovery of new therapeutic targets and the development of new treatment strategies for peripheral nerve regeneration. 展开更多
关键词 nerve regeneration peripheral nerve regeneration peripheral nerve injury sciatic nerve mi rnas mi R-30c dedifferentiation Schwann cells myelination in vivo in vitro neural regeneration
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捻转血矛线虫伊维菌素敏感虫株与耐药虫株差异miRNA的转录组学分析 被引量:5
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作者 温海峰 张艳敏 +10 位作者 张海龙 石雅琴 毛晓伟 寇慧琳 于宇 李峥 陈昕迪 王腾宇 高娃 王文龙 刘春霞 《中国预防兽医学报》 CAS CSCD 北大核心 2023年第3期245-252,共8页
为探究捻转血矛线虫的伊维菌素(IVM)敏感虫株与耐药虫株显著差异的microRNA(miRNA)转录谱,发掘其耐药相关的miRNA,本实验采用Illumina Hiseq2000平台分别对捻转血矛线虫IVM敏感虫株与耐药虫株测序,构建cDNA文库,通过质控后采用生物信息... 为探究捻转血矛线虫的伊维菌素(IVM)敏感虫株与耐药虫株显著差异的microRNA(miRNA)转录谱,发掘其耐药相关的miRNA,本实验采用Illumina Hiseq2000平台分别对捻转血矛线虫IVM敏感虫株与耐药虫株测序,构建cDNA文库,通过质控后采用生物信息学软件筛选捻转血矛线虫IVM敏感与耐药虫株的差异mi RNA。结果显示,捻转血矛线虫的IVM敏感与耐药虫株筛选出转录显著差异的miRNA共375个,其中253个转录上调,122个转录下调。利用RNAhybrid(V6.01)、Miranda(V3.3a)、TargetScan(V7.0)软件对筛选到的mi RNA靶基因进行预测,并取交集作为mi RNA靶基因预测结果,将筛选出显著差异的靶基因通过perl脚本与显著差异的miRNA进行关联分析,结果显示共关联到2106对miRNA-mRNA为负调控关系,其中涉及到477个差异的m RNA和603个差异的miRNA。对显著差异的miRNA靶基因进行GO富集分析,结果显示其中有492个GO terms,包括生物过程(BP)322个、细胞组分(CC)47个、分子功能(MF)123个。按照q值由小到大的顺序排列,被显著富集的有肌细胞发育分化及内肽酶活性等功能;按照富集基因数目由多到少排列,被显著富集的为有机物代谢、细胞代谢等功能。对显著差异的miRNA靶基因进行KEGG信号通路富集分析,按照q值由小到大的顺序排列,被显著富集到的有抗原处理与递呈等信号通路,其中显著差异靶基因也被富集到药物代谢-其他酶等与药物代谢相关的信号通路,以及一些在捻转血矛线虫耐IVM中起重要作用的通路,如:药物代谢细胞色素P450、细胞色素P450对外源药物代谢通路、ABC转运蛋白等。随机从差异miRNA中选10个进行RT-q PCR的验证,结果显示,相对转录水平结果与测序结果一致。本研究上述结果首次证实,捻转血矛线虫对IVM耐药性的产生与其代谢过程、肌细胞的分化发育过程及免疫过程中的某个或多个环节有一定的联系,本研究为进一步探究捻转血矛线虫耐药机制提供了科学依据。 展开更多
关键词 捻转血矛线虫 伊维菌素 耐药性 转录组 mi RNA
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Electroacupuncture induces acute changes in cerebral cortical mi RNA profile, improves cerebral blood flow and alleviates neurological deficits in a rat model of stroke 被引量:22
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作者 Hai-zhen Zheng Wei Jiang +6 位作者 Xiao-feng Zhao Jing Du Pan-gong Liu Li-dan Chang Wen-bo Li Han-tong Hu Xue-min Shi 《Neural Regeneration Research》 SCIE CAS CSCD 2016年第12期1940-1950,共11页
Electroacupuncture has been shown to improve cerebral blood flow in animal models of stroke. However, it is unclear whether electroacupuncture alters mi RNA expression in the cortex. In this study, we examined changes... Electroacupuncture has been shown to improve cerebral blood flow in animal models of stroke. However, it is unclear whether electroacupuncture alters mi RNA expression in the cortex. In this study, we examined changes in the cerebral cortical mi RNA profile, cerebral blood flow and neurological function induced by electroacupuncture in a rat model of stroke. Electroacupuncture was performed at Renzhong(GV26) and Neiguan(PC6), with a frequency of 2 Hz, continuous wave, current intensity of 3.0 m A, and stimulation time of 1 minute. Electroacupuncture increased cerebral blood flow and alleviated neurological impairment in the rats. mi RNA microarray profiling revealed that the vascular endothelial growth factor signaling pathway, which links cell proliferation with stroke, was most significantly affected by electroacupuncture. Electroacupuncture induced changes in expression of rno-mi R-206-3p, rno-mi R-3473, rno-mi R-6216 and rno-mi R-494-3p, and these changes were confirmed by quantitative real-time polymerase chain reaction. Our findings suggest that changes in cell proliferation-associated mi RNA expression induced by electroacupuncture might be associated with the improved cerebral blood supply and functional recovery following stroke. 展开更多
关键词 nerve regeneration stroke middle cerebral artery occlusion electroacupuncture mi RNA cerebral blood flow Neiguan(PC6) Renzhong(GV26) neural regeneration
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miRNA在阿霉素相关心脏毒性中调控作用的研究进展
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作者 董香军 郝长来 《生命科学研究》 CAS 2023年第4期290-297,329,共9页
心脏毒性是抗肿瘤药物阿霉素(doxorubicin,DOX)临床应用的主要限制之一,会促进严重心血管并发症的发展,这是一个亟待解决的健康问题。尽管经过了多年研究,但心脏毒性的机制仍然不清楚,也没有有效的早期预测或治疗方法。因此,需要进一步... 心脏毒性是抗肿瘤药物阿霉素(doxorubicin,DOX)临床应用的主要限制之一,会促进严重心血管并发症的发展,这是一个亟待解决的健康问题。尽管经过了多年研究,但心脏毒性的机制仍然不清楚,也没有有效的早期预测或治疗方法。因此,需要进一步了解心脏毒性的发生机制,以确保在心肌发生不可逆损伤之前制定早期预防或治疗策略。近年来,微RNA(microRNA,mi RNA)在阿霉素诱导的心脏毒性(doxorubicin-induced cardiotoxicity,DIC)中的作用引起了人们的广泛关注,mi RNA可能通过多种途径调控DIC的发生与发展,被认为是一个很有希望的探索领域。本文综述了DIC中与mi RNA相关的前沿研究,探讨了使用mi RNA作为治疗靶点的可能性和前景,并分析了其应用的局限性和挑战。 展开更多
关键词 阿霉素(DOX) 心脏毒性 微RNA(mi RNA) 氧化应激 线粒体损伤 自噬 凋亡
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The Biochemical Impact by Covalent Shielding of the Anionic Oxygen of the Phosphate Group in DNA and RNA as Methylated Phosphotriester Linkage on the Inhibition of DNA Duplication and on HIV-1 RNA Viral Infectivity Has Been Seriously Overlooked
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作者 Henk M. Buck 《Journal of Biophysical Chemistry》 CAS 2023年第2期59-66,共8页
With the help of model experiments, we are able to offer a detailed proposal for the inhibition of DNA duplication and no inhibition of RNA viral infectivity. As a backbone, we introduced methyl phosphotriester (MPTE)... With the help of model experiments, we are able to offer a detailed proposal for the inhibition of DNA duplication and no inhibition of RNA viral infectivity. As a backbone, we introduced methyl phosphotriester (MPTE). Duplex formation according to the traditional Watson and Crick base-pairing: [(MPTE)<sub>n−1</sub> DNA] * DNA and [(MPTE)<sub>n−1</sub> DNA] * RNA, where n = number of DNA and RNA bases. However, in the latter case, inhibition is obtained by reduction of the number of MPTE linkages, as is confirmed with model experiments and under biological conditions with micro (mi)RNA substrates. The latter results have recently been published. One or more single MPTEs are disseminated over different places of DNA without neighbour MPTEs (Prof. Wen-Yih Chen and his group, Taiwan). 展开更多
关键词 Methylated Phosphotriester (MPTE) DNA Partially MPTE DNA Model Inhibition Experiments micro (mi)RNA HIV-1 RNA Conformational Transition
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Absent MicroRNAs in Different Tissues of Patients with Acquired Cardiomyopathy
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作者 Christine S. Siegismund Maria Rohde +3 位作者 Uwe Kiihl Felicitas Escher Heinz Peter Schultheiss Dirk Lassner 《Genomics, Proteomics & Bioinformatics》 SCIE CAS CSCD 2016年第4期224-234,共11页
MicroRNAs (miRNAs) can be found in a wide range of tissues and body fluids, and their specific signatures can be used to determine diseases or predict clinical courses. The miRNA profiles in biological samples (tis... MicroRNAs (miRNAs) can be found in a wide range of tissues and body fluids, and their specific signatures can be used to determine diseases or predict clinical courses. The miRNA profiles in biological samples (tissue, serum, peripheral blood mononuelear cells or other body fluids) differ significantly even in the same patient and therefore have their own specificity for the presented condition. Complex profiles of deregulated miRNAs are of high interest, whereas the importance of non-expressed miRNAs was ignored. Since miRNAs regulate gene expression rather negatively, absent miRNAs could indicate genes with unaltered expression that therefore are normally expressed in specific compartments or under specific disease situations. For the first time, non-detectable miRNAs in different tissues and body fluids from patients with different diseases (cardiomyopathies, Alzheimer's disease, bladder cancer, and ocular cancer) were analyzed and compared in this study, miRNA expression data were generated by microarray or TaqMan PCR-based platforms. Lists of absent miRNAs of primarily cardiac patients (myocardium, blood cells, and serum) were clustered and analyzed for potentially involved pathways using two prediction platforms, i.e., miRNA enrichment analysis and annotation tool (miEAA) and DIANA miRPath. Extensive search in biomedical publication databases for the relevance of non-expressed miRNAs in predicted pathways revealed no evidence for their involvement in heart-related pathways as indicated by software tools, confirming proposed approach. 展开更多
关键词 CARDIOMYOPATHY Heart muscle biopsy Absent mi rnas Peripheral blood mononuclear cell Serum
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耐受性树突状细胞的研究进展 被引量:2
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作者 孙庐云 丁喆 +3 位作者 陈鹏 刘锋 宝福凯 胡明道 《生命科学研究》 CAS CSCD 2020年第4期314-320,共7页
耐受性树突状细胞(tolerogenic dendritic cell, tolDC)在器官移植和自体免疫性疾病中对免疫耐受调节至关重要。与免疫抑制剂相比, tolDC用于治疗移植后排斥反应的副作用较低。此外, tolDC可通过多种途径培养生成,且不同的培养方式具有... 耐受性树突状细胞(tolerogenic dendritic cell, tolDC)在器官移植和自体免疫性疾病中对免疫耐受调节至关重要。与免疫抑制剂相比, tolDC用于治疗移植后排斥反应的副作用较低。此外, tolDC可通过多种途径培养生成,且不同的培养方式具有独特功效。目前, tolDC作为一种诱导器官移植耐受的辅助治疗已在临床试验中广泛应用。本文就tolDC诱导免疫耐受、tolDC的体外诱导培养及其mi RNA调控机制和应用进行综述并提出相关展望,以期为应用tolDC治疗减轻移植后排斥反应的研究提供新思路。 展开更多
关键词 耐受性树突状细胞(tolDC) 免疫耐受 移植免疫 移植排斥 mi RNA调控机制
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The role of circulating miRNAs in multiple myeloma 被引量:4
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作者 ZHANG Ji XIAO XiaoJuan LIU Jing 《Science China(Life Sciences)》 SCIE CAS CSCD 2015年第12期1262-1269,共8页
Multiple myeloma(MM) is a common malignant hematological disease. Dysregulation of micro RNAs(mi RNAs) in MM cells and bone marrow microenviroment has important impacts on the initiation and progression of MM and drug... Multiple myeloma(MM) is a common malignant hematological disease. Dysregulation of micro RNAs(mi RNAs) in MM cells and bone marrow microenviroment has important impacts on the initiation and progression of MM and drug resistance in MM cells. Recently, it was reported that MM patient serum and plasma contained sufficiently stable mi RNA signatures, and circulating mi RNAs could be identified and measured accurately from body fluid. Compared to conventional diagnostic parameters, the circulating mi RNA profile is appropriate for the diagnosis of MM and estimates patient progression and therapeutic outcome with higher specificity and sensitivity. In this review, we mainly focus on the potential of circulating mi RNAs as diagnostic, prognostic, and predictive biomarkers for MM and summarize the general strategies and methodologies for identification and measurement of circulating mi RNAs in various cancers. Furthermore, we discuss the correlation between circulating mi RNAs and the cytogenetic abnormalities and biochemical parameters assessed in multiple myeloma. 展开更多
关键词 mi RNA multiple myeloma BIOMARKER DIAGNOSIS
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CircLphn3 protects the blood-brain barrier in traumatic brain injury 被引量:3
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作者 Yu-Qi Cheng Chen-Rui Wu +6 位作者 Meng-Ran Du Qiang Zhou Bi-Ying Wu Jia-Yuan-Yuan Fu Ehab Balawi Wei-Lin Tan Zheng-Bu Liao 《Neural Regeneration Research》 SCIE CAS CSCD 2022年第4期812-818,共7页
Circular RNAs(circRNAs)are a new and large group of non-coding RNA molecules that are abundantly expressed in the central nervous system.However,very little is known about their roles in traumatic brain injury.In this... Circular RNAs(circRNAs)are a new and large group of non-coding RNA molecules that are abundantly expressed in the central nervous system.However,very little is known about their roles in traumatic brain injury.In this study,we firstly screened differentially expressed circ RNAs in normal and injured brain tissues of mice after traumatic brain injury.We found that the expression of circ Lphn3 was substantially decreased in mouse models of traumatic brain injury and in hemin-treated b End.3(mouse brain cell line)cells.After overexpressing circ Lphn3 in b End.3 cells,the expression of the tight junction proteins,ZO-1,ZO-2,and occludin,was upregulated,and the expression of mi R-185-5 p was decreased.In b End.3 cells transfected with mi R-185-5 p mimics,the expression of ZO-1 was decreased.Dual-luciferase reporter assays showed that circ Lphn3 bound to mi R-185-5 p,and that mi R-185-5 p bound to ZO-1.Additionally,circ Lphn3 overexpression attenuated the hemin-induced high permeability of the in vitro b End.3 cell model of the blood-brain barrier,while mi R-185-5 p transfection increased the permeability.These findings suggest that circ Lphn3,as a molecular sponge of mi R-185-5 p,regulates tight junction proteins'expression after traumatic brain injury,and it thereby improves the permeability of the blood-brain barrier.This study was approved by the Animal Care and Use Committee of Chongqing Medical University of China(approval No.2021-177)on March 22,2021. 展开更多
关键词 BIOINFORMATICS blood-brain barrier circ RNA mi RNA RNA-sequence tight junction protein traumatic brain injury ZO1
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小RNA高通量测序数据分析方法 被引量:1
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作者 彭骅 李佛生 王胜华 《生命科学仪器》 2017年第2期19-28,共10页
本文应用Perl语言和My SQL数据库构建了小RNA高通量测序数据分析平台,以4个水稻数据集为分析对象,详细介绍了小RNA高通量测序数据的处理方法和流程。我们以MSU 6.1水稻基因组为参考,构建了该版本的全基因组结构及已知nc RNAs位点信息数... 本文应用Perl语言和My SQL数据库构建了小RNA高通量测序数据分析平台,以4个水稻数据集为分析对象,详细介绍了小RNA高通量测序数据的处理方法和流程。我们以MSU 6.1水稻基因组为参考,构建了该版本的全基因组结构及已知nc RNAs位点信息数据库,结合Perl脚本可以实现小RNA在基因组上的详细定位与统计,同时我们从数据库中提取已知pre-mi RNAs表达特征,设计了一个新的mi RNAs挖掘方法,该方法可以筛选出大量的新mi RNAs,其中已知mi RNAs命中率可以达到98%。针对水稻小RNA种类的多样性,我们对mi RNAs和endo-si RNAs的鉴别也给予了探讨和说明。本文设计的高通量测序数据分析平台,方法简单高效,以数据库作为存储和查询媒介,能够实现多位点reads的分析,可以得到灵活多样的数据统计结果。依照本文的方法同样可以构建其他模式物种的小RNA数据分析平台,在高通量测序逐渐普及的将来,本文的方法对中小实验室建立自己的数据分析平台具有实践指导意义。 展开更多
关键词 小RNA(small rnas) 非编码RNA(nc rnas) 微小RNA(micro rnas mi rnas)内源小干扰RNA(endo-si rnas) 小RNA高通量测序(small RNA-Seq)
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靛玉红通过miRNA-29b-3p/TET2/5hmC轴促进骨髓增生异常综合征细胞株SKM-1细胞DNA去甲基化
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作者 喻丽 凌志明 +3 位作者 谷晓丽 杨秀鹏 陈朋杰 许勇钢 《中华中医药杂志》 CAS CSCD 北大核心 2024年第10期5448-5452,共5页
目的:探索青黄散有效成分靛玉红对骨髓增生异常综合征(MDS)的去甲基化作用机制。方法:以MDS细胞株SKM-1细胞为研究对象,以不同浓度的靛玉红干预细胞48 h,采用DNA斑点印记方法,检测5-羟甲基胞嘧啶(5hmC)、5-甲基胞嘧啶(5mC)等DNA甲基化... 目的:探索青黄散有效成分靛玉红对骨髓增生异常综合征(MDS)的去甲基化作用机制。方法:以MDS细胞株SKM-1细胞为研究对象,以不同浓度的靛玉红干预细胞48 h,采用DNA斑点印记方法,检测5-羟甲基胞嘧啶(5hmC)、5-甲基胞嘧啶(5mC)等DNA甲基化指标的变化情况;通过qPCR、蛋白免疫印迹检测SKM-1细胞TET2基因表达;通过转录组高通量测序分析靛玉红调控SKM-1细胞miRNA表达的情况,并通过miRNA靶基因预测工具miranda工具筛选参与靛玉红促进MDS的TET2表达的候选miRNAs,随后使用qPCR进行实验验证。结果:DNA斑点印记结果显示靛玉红显著提高了SKM-1细胞DNA中5hmC的水平而降低了5mC的表达水平。qPCR与蛋白免疫印迹结果显示靛玉红没有显著改变TET2 mRNA表达,但是显著提高了TET2蛋白表达。转录组高通量测序结果显示靛玉红干预后有35个miRNA存在表达差异,其中17个miRNA上调,18个miRNA下调。Miranda结果显示差异miRNA中miRNA-29b-3p、miRNA-125a-5p可能作用于TET2基因。qPCR结果显示,靛玉红下调了miRNA-29b-3p的表达而上调了mi RNA-125a-5p的表达。结论:靛玉红可能通过mi RNA-29b-3p/TET2/5hm C轴促进SKM-1细胞DNA去甲基化。 展开更多
关键词 骨髓增生异常综合征 SKM-1细胞 DNA甲基化 靛玉红 TET2 mi RNA
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MicroRNA与多囊卵巢综合征关系的研究进展 被引量:2
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作者 潘乐乐 肖国宏 杨洁 《生殖与避孕》 CAS CSCD 北大核心 2015年第4期265-270,共6页
Micro RNAs(mi RNA)是一类长度为19-25nt的内源性非编码RNA,主要参与调节基因转录后水平的表达。研究发现mi RNAs在卵巢组织中广泛表达,在卵巢功能调节过程中发挥着重要的作用。近年来,越来越多的证据表明,mi RNAs与多囊卵巢综合征(P... Micro RNAs(mi RNA)是一类长度为19-25nt的内源性非编码RNA,主要参与调节基因转录后水平的表达。研究发现mi RNAs在卵巢组织中广泛表达,在卵巢功能调节过程中发挥着重要的作用。近年来,越来越多的证据表明,mi RNAs与多囊卵巢综合征(PCOS)关系密切。mi RNA表达水平的变化与PCOS患者甾体激素调节紊乱、高雄激素血症、胰岛素抵抗及不孕症等主要表现有关。将来,mi RNAs有望成为PCOS疾病诊断和预后评估的潜在生物学标志物,可为PCOS的治疗提供新思路。 展开更多
关键词 micro RNA(mi RNA) 多囊卵巢综合征(PCOS) 高雄激素血症 胰岛素抵抗(IR) 不孕症
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Comparative studies of two methods for miRNA isolation from milk whey 被引量:1
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作者 Xiao-lu JIN Zi-hai WEI +2 位作者 Lan LIU Hong-yun LIU Jian-xin LIU 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2015年第6期533-540,共8页
Micro RNAs(mi RNAs) from milk whey have been considered for their potential as noninvasive biomarkers for milk quality control and disease diagnosis. However, standard protocols for mi RNA isolation and quantificati... Micro RNAs(mi RNAs) from milk whey have been considered for their potential as noninvasive biomarkers for milk quality control and disease diagnosis. However, standard protocols for mi RNA isolation and quantification from milk whey are not well established. The objective of this study was to compare two methods for the isolation of mi RNAs from milk whey. These two methods were modified phenol-based technique(Trizol LS followed by phenol precipitation, the TP method) and combined phenol and column-based approach(Trizol LS followed by cleanup using the mi RNeasy kit, the TM method). Yield and quality of RNA were rigorously measured using a Nano Drop ND-1000 spectrophotometer and then the distribution of RNA was precisely detected in a Bioanalyzer 2100 instrument by microchip gel electrophoresis. Several endogenous mi RNAs(bta-mi R-141, bta-mi R-146 a, bta-mi R-148 a, bta-mi R-200 c, bta-mi R-362, and bta-mi R-375) and an exogenous spike-in synthetic control mi RNA(cel-mi R-39) were quantified by real-time polymerase chain reaction(PCR) to examine the apparent recovery efficiency of milk whey mi RNAs. Both methods could successfully isolate sufficient small RNA(200 nt) from milk whey, and their yields were quite similar. However, the quantification results show that the total mi RNA recovery efficiency by the TM method is superior to that by the TP method. The TM method performed better than the TP for recovery of milk whey mi RNA due to its consistency and good repeatability in endogenous and spike-in mi RNA recovery. Additionally, quantitative recovery analysis of a spike-in mi RNA may be more accurate to reflect the milk whey mi RNA recovery efficiency than using traditional RNA quality analysis instruments(Nano Drop or Bioanalyzer 2100). 展开更多
关键词 Method milk whey mi RNA Spike-in miRNA
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