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Selective separation and enrichment of proteins in aqueous two-phase extraction system
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作者 Feng Qu Hao Qin +3 位作者 Min Dong Dong Xu Zhao Xin Ying Zhao Jing Hua Zhang 《Chinese Chemical Letters》 SCIE CAS CSCD 2009年第9期1100-1102,共3页
A simple aqueous two-phase extraction system (ATPS) of PEG/phosphate was proposed for selective separation and enrichment of proteins. The combination of ATPE with HPLC was applied to identify the partition of prote... A simple aqueous two-phase extraction system (ATPS) of PEG/phosphate was proposed for selective separation and enrichment of proteins. The combination of ATPE with HPLC was applied to identify the partition of proteins in two phases. Five proteins (bovine serum albumin, Cytochrome C, lysozyme, myoglobin, and trypsin) were used as model proteins to study the effect of phosphate concentration and pH on proteins partition. The PEG/phosphate system was firstly applied to real human saliva and plasma samples, some proteins showed obviously different partition in two phases. The primary results manifest the selective separation and enrichment of proteins in ATPS provided the potential for high abundance proteins depletion in proteomics. ~ 2009 Feng Qu. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved. 展开更多
关键词 Aqueous two-phase extraction Saliva and plasma proteinS Selective separation and enrichment
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Protein Enrichment of Potato Peels Using Saccharomyces cerevisiae via Solid-State Fermentation Process
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作者 Onoh Ikechukwu Maxwell Udeh Benson Chinwuba Mbah Gordian Onyebuchukwu 《Advances in Chemical Engineering and Science》 2019年第1期99-108,共10页
In order to add value to potato peels and also curb their environmental pollution problems, this study investigated the protein enrichment of potato peels with Saccharomyces cerevisiae via Solid-State Fermentation (SS... In order to add value to potato peels and also curb their environmental pollution problems, this study investigated the protein enrichment of potato peels with Saccharomyces cerevisiae via Solid-State Fermentation (SSF). SSF is a fermentation process which involves solid matrix and is carried out in absence or near absence of free water. SSF of potato peel mashed was carried out with S. cerevisiae at 30°C, pH of 5.5, moisture adjustment between 40 and 90%, addition of ammonium sulphate and urea salts as nitrogen supplements for the microorganisms for 3 days. The results showed that the percentage crude protein content of all the fermented samples increased significantly when compared with the unfermented sample. 40% moisture content adjustment and ammonium sulphate as nitrogen source gave the best result. The crude protein increased from 12.5% to 21.86%, which is 74.88% increment for ammonium sulphate supplementation, and 12.5% to 18.42%, which is 47% increment for urea supplementation. Therefore, the fermented peels could serve as good source of cheap protein enriched feed for livestock. 展开更多
关键词 POTATO Peel SACCHAROMYCES cerevisiae Urea Ammonium Sulphate Crude Pro-tein SOLID-STATE Fermentation protein enrichment AOAC 1990
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An enriched environment increases the expression of fibronectin type Ⅲ domain-containing protein 5 and brain-derived neurotrophic factor in the cerebral cortex of the ischemic mouse brain 被引量:12
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作者 Ke-Wei Yu Chuan-Jie Wang +7 位作者 Yi Wu Yu-Yang Wang Nian-Hong Wang Shen-Yi Kuang Gang Liu Hong-Yu Xie Cong-Yu Jiang Jun-Fa Wu 《Neural Regeneration Research》 SCIE CAS CSCD 2020年第9期1671-1677,共7页
Many studies have shown that fibronectin type III domain-containing protein 5(FDNC5) and brain-derived neurotrophic factor(BDNF) play vital roles in plasticity after brain injury. An enriched environment refers to an ... Many studies have shown that fibronectin type III domain-containing protein 5(FDNC5) and brain-derived neurotrophic factor(BDNF) play vital roles in plasticity after brain injury. An enriched environment refers to an environment that provides animals with multi-sensory stimulation and movement opportunities. An enriched environment has been shown to promote the regeneration of nerve cells, synapses, and blood vessels in the animal brain after cerebral ischemia;however, the exact mechanisms have not been clarified. This study aimed to determine whether an enriched environment could improve neurobehavioral functions after the experimental inducement of cerebral ischemia and whether neurobehavioral outcomes were associated with the expression of FDNC5 and BDNF. This study established ischemic mouse models using permanent middle cerebral artery occlusion(pMCAO) on the left side. On postoperative day 1, the mice were randomly assigned to either enriched environment or standard housing condition groups. Mice in the standard housing condition group were housed and fed under standard conditions. Mice in the enriched environment group were housed in a large cage, containing various toys, and fed with a standard diet. Sham-operated mice received the same procedure, but without artery occlusion, and were housed and fed under standard conditions. On postoperative days 7 and 14, a beam-walking test was used to assess coordination, balance, and spatial learning. On postoperative days 16–20, a Morris water maze test was used to assess spatial learning and memory. On postoperative day 15, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex were analyzed by western blot assay. The results showed that compared with the standard housing condition group, the motor balance and coordination functions(based on beam-walking test scores 7 and 14 days after operation), spatial learning abilities(based on the spatial learning scores from the Morris water maze test 16–19 days after operation), and memory abilities(based on the memory scores of the Morris water maze test 20 days after operation) of the enriched environment group improved significantly. In addition, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex increased in the enriched environment group compared with those in the standard housing condition group. Furthermore, the Pearson correlation coefficient showed that neurobehavioral functions were positively associated with the expression levels of FDNC5 and BDNF(r = 0.587 and r = 0.840, respectively). These findings suggest that an enriched environment upregulates FDNC5 protein expression in the ipsilateral cerebral cortex after cerebral ischemia, which then activates BDNF protein expression, improving neurological function. BDNF protein expression was positively correlated with improved neurological function. The experimental protocols were approved by the Institutional Animal Care and Use Committee of Fudan University, China(approval Nos. 20160858 A232, 20160860 A234) on February 24, 2016. 展开更多
关键词 beam-walking test brain-derived neurotrophic factor cerebral ischemia correlation analysis enriched environment fibronectin typeⅢdomain-containing protein 5 Morris water maze task neural plasticity NEUROPROTECTION permanent middle cerebral artery occlusion
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Environmental enrichment combined with fasudil promotes motor function recovery and axonal regeneration after stroke 被引量:8
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作者 Yi-Tong Zhu Qun Zhang +4 位作者 Hong-Yu Xie Ke-Wei Yu Gao-Jing Xu Si-Yue Li Yi Wu 《Neural Regeneration Research》 SCIE CAS CSCD 2021年第12期2512-2520,共9页
Fasudil,a Rho-associated protein kinase(ROCK)inhibitor,has a protective effect on the central nervous system.In addition,environmental enrichment is a promising technique for inducing the recovery of motor impairments... Fasudil,a Rho-associated protein kinase(ROCK)inhibitor,has a protective effect on the central nervous system.In addition,environmental enrichment is a promising technique for inducing the recovery of motor impairments in ischemic stroke models.The present study aimed to explore whether environmental enrichment combined with fasudil can facilitate motor function recovery and induce cortical axonal regeneration after stroke.First,a mouse model of ischemic cerebral stroke was established by photochemical embolization of the left sensorimotor cortex.Fasudil solution(10 mg/kg per day)was injected intraperitoneally for 21 days after the photothrombotic stroke.An environmental enrichment intervention was performed on days 7-21 after the photothrombotic stroke.The results revealed that environmental enrichment combined with fasudil improved motor function,increased growth-associated protein 43 expression in the infarcted cerebral cortex,promoted axonal regeneration on the contralateral side,and downregulated ROCK,p-LIM domain kinase(LIMK)1,and p-cofilin expression.The combined intervention was superior to monotherapy.These findings suggest that environmental enrichment combined with fasudil treatment promotes motor recovery after stroke,at least partly by stimulating axonal regeneration.The underlying mechanism might involve ROCK/LIMK1/cofilin pathway regulation.This study was approved by the Institutional Animal Care and Use Committee of Fudan University,China(approval No.20160858A232)on February 24,2016. 展开更多
关键词 axon regeneration biotinylated dextran amines environmental enrichment FASUDIL growth-associated protein 43 ischemic stroke motor recovery Nissl bodies Rho/ROCK pathway
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Axon regeneration induced by environmental enrichment-epigenetic mechanisms 被引量:1
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作者 Bor Luen Tang 《Neural Regeneration Research》 SCIE CAS CSCD 2020年第1期10-15,共6页
Environmental enrichment is known to be beneficial for cognitive improvement.In many animal models of neurological disorders and brain injury,EE has also demonstrated neuroprotective benefits in neurodegenerative dise... Environmental enrichment is known to be beneficial for cognitive improvement.In many animal models of neurological disorders and brain injury,EE has also demonstrated neuroprotective benefits in neurodegenerative diseases and in improving recovery after stroke or traumatic brain injury.The exact underlying mechanism for these phenomena has been unclear.Recent findings have now indicated that neuronal activity elicited by environmental enrichment induces Ca2+influx in dorsal root ganglion neurons results in lasting enhancement of CREB-binding protein-mediated histone acetylation.This,in turn,increases the expression of pro-regeneration genes and promotes axonal regeneration.This mechanism associated with neuronal activity elicited by environmental enrichment-mediated pathway is one of several epigenetic mechanisms which modulate axon regeneration upon injury that has recently come to light.The other prominent mechanisms,albeit not yet directly associated with environmental enrichment,include DNA methylation/demethylation and N6-methyladenosine modification of transcripts.In this brief review,I highlight recent work that has shed light on the epigenetic basis of environmental enrichment-based axon regeneration,and discuss the mechanism and pathways involved.I further speculate on the implications of the findings,in conjunction with the other epigenetic mechanisms,that could be harness to promote axon regeneration upon injury. 展开更多
关键词 AXON regeneration CREB-binding protein DNA methylation/demethylation dorsal root GANGLION DRG neurons environmental enrichment epigenetics histone acetylation mechanistic target of rapamycin mTOR PHOSPHATASE and TENSIN HOMOLOGUE PTEN
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Combining Phytate/Ca^(2+) Fractionation with Trichloroacetic Acid/Acetone Precipitation Improved Separation of Low-Abundant Proteins of Wheat (Triticum aestivum L.) Leaf for Proteomic Analysis 被引量:1
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作者 Muhammad A R F Sultan LIU Hui +2 位作者 CHENG Yu-Feng ZHANG Pei-pei ZHAO Hui-xian 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2013年第7期1123-1129,共7页
Proteomic assessment of low-abundance leaf proteins is hindered by the large quantity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) present within plant leaf tissues. In the present study, total prote... Proteomic assessment of low-abundance leaf proteins is hindered by the large quantity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) present within plant leaf tissues. In the present study, total proteins were extracted from wheat (Triticum aestivum L.) leaves by a conventional trichloroacetic acid (TCA)/acetone method and a protocol first developed in this work. Phytate/Ca2+ fractionation and TCA/acetone precipitation were combined to design an improved TCA/acetone method. The extracted proteins were analysed by two-dimensional gel electrophoresis (2-DE). The resulting 2-DE images were compared to reveal major differences. The results showed that large quantities of Rubisco were deleted from wheat leaf proteins prepared by the improved method. As many as (758±4) protein spots were detected from 2-DE images of protein extracts obtained by the improved method, 130 more than those detected by the TCA/acetone method. Further analysis indicated that more protein spots could be detected at regions of pI 4.00-4.99 and 6.50-7.00 in the improved method-based 2-DE images. Our findings indicated that the improved method is an efficient protein preparation protocol for separating low-abundance proteins in wheat leaf tissues by 2-DE analysis. The proposed protocol is simple, fast, inexpensive and also applicable to protein preparations of other plants. 展开更多
关键词 Triticum aestivum L. RUBISCO low-abundance protein phytate/Ca2+ two-dimensional gel electrophoresis plant leaf proteomics
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Optimization and Quality Characters of Protein Enriched Pasta
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作者 A. Kaur S. Sharma H. P. S. Nagi G. Kaur 《Journal of Food Science and Engineering》 2011年第4期271-281,共11页
The response surface methodology (Box-behnken design) was acknowledged for the production of optimized protein enriched pasta. Studied responses (protein, overall acceptability, hue angle and hardness) resulted to... The response surface methodology (Box-behnken design) was acknowledged for the production of optimized protein enriched pasta. Studied responses (protein, overall acceptability, hue angle and hardness) resulted to high protein pasta. The optima were at 12.5%,5 mung bean, 10% whey protein concentrate and 6% egg albumen in combination. Best combination of protein source and their respective level for high protein pasta on the quality evaluation basis were mung bean (15%) + whey protein concentrate (10%); mung bean (15%) + egg albumen (6%); mung bean (15%) + whey protein concentrate (10%) + egg albumen (6%). Hue angle and Hardness remained within acceptable range. Model adequacy was supported with lack of fit test. Enriched protein pasta had excellent cooking and sensory quality during storage period of 4 months. ERH value ranged from 34-36 per cent had supported the results. Rheological properties and good storage behaviour (4 months) had also enhanced the quality of optimized protein enriched pasta. Enriched pasta was rated excellent by 70 per cent of the consumers in mass acceptability trials. 展开更多
关键词 PASTA protein enrichment RSM Box-behnken mung bean flour whey protein concentrate egg albumen.
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Comprehensive and deep profiling of the plasma proteome with protein corona on zeolite NaY
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作者 Congcong Ma Yanwei Li +10 位作者 Jie Li Lei Song Liangyu Chen Na Zhao Xueping Li Ning Chen Lixia Long Jin Zhao Xin Hou Li Ren Xubo Yuan 《Journal of Pharmaceutical Analysis》 SCIE CAS CSCD 2023年第5期503-513,共11页
Proteomic characterization of plasma is critical for the development of novel pharmacodynamic biomarkers.However,the vast dynamic range renders the profiling of proteomes extremely challenging.Here,we synthesized zeol... Proteomic characterization of plasma is critical for the development of novel pharmacodynamic biomarkers.However,the vast dynamic range renders the profiling of proteomes extremely challenging.Here,we synthesized zeolite NaY and developed a simple and rapid method to achieve comprehensive and deep profiling of the plasma proteome using the plasma protein corona formed on zeolite NaY.Specifically,zeolite NaY and plasma were co-incubated to form plasma protein corona on zeolite NaY(NaY-PPC),followed by conventional protein identification using liquid chromatography-tandem mass spectrometry.NaY was able to significantly enhance the detection of low-abundance plasma proteins,minimizing the“masking”effect caused by high-abundance proteins.The relative abundance of middleand low-abundance proteins increased substantially from 2.54%to 54.41%,and the top 20 highabundance proteins decreased from 83.63%to 25.77%.Notably,our method can quantify approximately 4000 plasma proteins with sensitivity up to pg/mL,compared to only about 600 proteins identified from untreated plasma samples.A pilot study based on plasma samples from 30 lung adenocarcinoma patients and 15 healthy subjects demonstrated that our method could successfully distinguish between healthy and disease states.In summary,this work provides an advantageous tool for the exploration of plasma proteomics and its translational applications. 展开更多
关键词 NAY Plasma proteomics protein corona low-abundance proteins
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不同富硒方式对甘薯叶蛋白抗氧化活性的影响
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作者 高琦 段文瀚 +4 位作者 彭雪 王宁 卢雅婷 张倩 薛友林 《食品科学》 EI CAS CSCD 北大核心 2024年第9期29-35,共7页
以甘薯叶为研究对象,分别对叶面与土壤进行富硒处理,通过分析甘薯叶中硒含量及甘薯叶蛋白的抗氧化活性的变化,研究不同富硒方式对甘薯叶蛋白抗氧化活性的影响。结果显示叶面富硒与土壤富硒都能提高甘薯叶蛋白的硒含量,在1.6 mg/mL的富... 以甘薯叶为研究对象,分别对叶面与土壤进行富硒处理,通过分析甘薯叶中硒含量及甘薯叶蛋白的抗氧化活性的变化,研究不同富硒方式对甘薯叶蛋白抗氧化活性的影响。结果显示叶面富硒与土壤富硒都能提高甘薯叶蛋白的硒含量,在1.6 mg/mL的富硒质量浓度条件下两种富硒处理组蛋白硒含量分别达到了20.10 mg/g与1.68 mg/g。富硒对甘薯叶蛋白的抗氧化活性有一定的提升作用,其中,叶面富硒可使甘薯叶蛋白超氧阴离子自由基清除率提升19.17%。根据相关性分析可知,除硒含量对抗氧化活性有较大影响外,样品中总酚与总黄酮含量也在一定程度上影响着样品的抗氧化活性。经对比可知,叶面富硒能更好地提高甘薯叶蛋白中的硒含量,是较佳的富硒方式。 展开更多
关键词 富硒 甘薯叶蛋白 抗氧化活性
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蛋白质N-磷酸化修饰富集方法进展 被引量:1
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作者 江波 高博 +3 位作者 魏淑娴 梁振 张丽华 张玉奎 《色谱》 CAS CSCD 北大核心 2024年第7期623-631,共9页
蛋白质磷酸化作为一种最普遍和最重要的翻译后修饰调控着几乎所有的生命过程。随着高效富集方法和生物质谱技术的快速发展,低丰度的蛋白质O-磷酸化修饰获得了规模化鉴定,从而使其生物学功能得到较为透彻的研究。而发生在组氨酸、赖氨酸... 蛋白质磷酸化作为一种最普遍和最重要的翻译后修饰调控着几乎所有的生命过程。随着高效富集方法和生物质谱技术的快速发展,低丰度的蛋白质O-磷酸化修饰获得了规模化鉴定,从而使其生物学功能得到较为透彻的研究。而发生在组氨酸、赖氨酸和精氨酸侧链氨基的N-磷酸化修饰,由于P-N键化学稳定性差,导致其在酸和热条件下不稳定。而目前依赖酸性条件的O-磷酸化富集方法难以适用N-磷酸化富集,导致蛋白质N-磷酸化生物功能研究严重滞后。因此,迫切需要发展针对蛋白质N-磷酸化的高效富集方法。本文首先介绍了蛋白质N-磷酸化的结构特征和已报道的生物学功能,重点综述并分析了近20年来蛋白质N-磷酸化修饰富集方法,并对每一种富集方法的优缺点进行了评述,最后对潜在的富集方法进行了展望。 展开更多
关键词 蛋白质N-磷酸化 生物功能 富集方法 综述
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富硒小米蛋白的理化性质、功能特性及结构研究 被引量:1
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作者 李琳 金丽娜 +2 位作者 郭佩佩 刘敦华 傅婧 《食品与发酵工业》 CAS CSCD 北大核心 2024年第3期259-267,共9页
该文以普通小米蛋白(common millet protein,MP)为对照,采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)、傅里叶红外光谱(Fourier infrared spectroscopy,FT-IR)、扫描电... 该文以普通小米蛋白(common millet protein,MP)为对照,采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)、傅里叶红外光谱(Fourier infrared spectroscopy,FT-IR)、扫描电镜(scanning electron microscopy,SEM)等方法并测定溶解度(solubility,PS)、持水性(water holding capacity,WA)、乳化性(emulsification capability,EC)等指标对富硒小米蛋白(selenium-rich millet protein,SMP)的理化性质、功能特性及结构进行分析。结果表明,MP与SMP的硒含量分别为0.08、0.63 mg/kg;MP与SMP的必需氨基酸(essential amino acid,EAA)含量分别占总氨基酸含量的51.101%、53.529%;EAA与非必需氨基酸(non-essential amino acids,NEAA)的比值为1.045、1.152;2种小米蛋白分子质量均分布在10~70.2 kDa;随着pH升高,小米蛋白的PS、WA、起泡性(froth capability,FC)、起泡稳定性(froth stability,FS)、EC均呈现先下降后上升趋势,而乳化稳定性(emulsion stability,ES)正好相反;随着温度升高,小米蛋白的PS、WA、持油性(oil-holding property,FA)、FC、FS、EC、ES均呈现先上升后下降趋势(P<0.05);SEM结果显示MP表面有不规则凸起,而SMP表面光滑平整;FT-IR显示2种小米蛋白峰型一致,SMP位置略有后移,同时SMP的α-螺旋、β-折叠、β-转角含量较MP分别高出5.532、2.885、5.506个百分点,无规则卷曲含量则低于MP,说明SMP的结构有序性优于MP。该研究结果为SMP产品的开发利用提供重要理论依据。 展开更多
关键词 富硒小米蛋白 功能特性 理化特性 结构表征
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海马G蛋白偶联雌激素受体1参与癫痫调控的转录组学研究 被引量:1
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作者 左娣 郝世杰 +8 位作者 杨盼 李苗 任晓璠 丁娜 马文倩 王盼盼 王诗雨 戎伟芳 刘昆梅 《中国药理学通报》 CAS CSCD 北大核心 2024年第4期709-715,共7页
目的本研究分别将野生型(WT)、Gper1敲低(Gper1-KD)大鼠海马组织进行转录组测序,探讨G蛋白偶联雌激素受体1(G-protein coupled estrogen receptor 1,GPER1)影响癫痫发病可能的信号通路和分子机制。方法海马组织进行RNA提取和cDNA文库构... 目的本研究分别将野生型(WT)、Gper1敲低(Gper1-KD)大鼠海马组织进行转录组测序,探讨G蛋白偶联雌激素受体1(G-protein coupled estrogen receptor 1,GPER1)影响癫痫发病可能的信号通路和分子机制。方法海马组织进行RNA提取和cDNA文库构建,与NCBI数据库大鼠基因组及基因注释比对,通过FPKM值,筛选组别间差异表达基因(DEGs)。将DEGs进行GO富集、KEGG富集分析,构建蛋白互作网络,利用RT-qPCR法对关键DEGs进行验证。结果Gper1-KD组与WT组相比,筛选(Fold change>2且FDR<0.01)后,检测到DEGs 2253个,其中上调基因1380个,下调基因873个;GO结果显示,DEGs主要分布在淋巴细胞趋化性、细胞分泌、巨噬细胞趋化性、中性粒细胞趋化性、血管生成正向调控等生物过程;KEGG结果显示,DEGs相关分子信号通路主要有癌症信号通路、PI3K-Akt通路、癌症蛋白聚糖相关信号通路、细胞黏附相关通路、MAPK信号通路等;RT-qPCR结果表明,Mapk12、Pdpk1、Foxo3、Camk2d、Pik3cg等基因表达水平差异具有显著性。结论MAPK、TNF信号通路在GPER1影响癫痫发生中可能起关键作用,GABA能神经元和Pik3cg在GPER1影响癫痫易感性方面可能发挥重要作用。 展开更多
关键词 GPER1 癫痫 DEGs GO富集 KEGG富集 蛋白互作网络
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芥菜SRO基因家族全基因组鉴定与表达分析
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作者 杨巍 赵丽芬 +7 位作者 唐兵 周麟笔 杨娟 莫传园 张宝会 李飞 阮松林 邓英 《生物技术通报》 CAS CSCD 北大核心 2024年第8期129-141,共13页
【目的】SRO基因家族是植物特有的一类转录因子,在植物生长发育与胁迫响应中具有重要作用,研究芥菜SRO基因家族,为解析芥菜SRO基因功能与遗传改良提供理论依据。【方法】利用生物信息学鉴定油用芥菜与菜用芥菜基因组中的SRO基因家族成员... 【目的】SRO基因家族是植物特有的一类转录因子,在植物生长发育与胁迫响应中具有重要作用,研究芥菜SRO基因家族,为解析芥菜SRO基因功能与遗传改良提供理论依据。【方法】利用生物信息学鉴定油用芥菜与菜用芥菜基因组中的SRO基因家族成员,运用TBtools、MEGA、Cytoscape、NCBI、STRING、EggNOG等软件与数据库进行理化性质、序列特征、进化关系、调控网络等分析以及RT-qPCR分析盐胁迫下的表达模式。【结果】两种类型芥菜SRO基因家族成员数量存在差异,并分属A与B两大类,其中A类SRO蛋白含有WWE、PARP及RST结构域,而B类蛋白则缺少WWE结构域,SRO基因启动子区域含有多种非生物胁迫响应与激素响应的顺式作用元件,microRNA-BjuSRO-靶基因构建复杂调控网络,参与了细胞凋亡、根系形态建成、免疫应答、异源刺激的细胞反应、对活性氧的反应等生物学过程,油用芥菜BjuOSRO基因与甘蓝SRO基因具有较近的进化关系,RT-qPCR结果显示,在盐胁迫下BjuVA1a、BjuVA1e、BjuVA2a、BjuVA3a、BjuVB1b、BjuVA2a显著上调表达。【结论】芥菜SRO基因家族具有功能多样性,BjuVA1a、BjuVA1e、BjuVA2a、BjuVA3a、BjuVB1b、BjuVA2a与盐胁迫响应密切相关,可作为培育耐盐型芥菜新品种的候选基因。 展开更多
关键词 芥菜 SRO转录因子 生物信息学 MICRORNA调控 蛋白互作 GO富集分析 进化分析 基因表达分析
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静脉注射利多卡因后差异蛋白质的蛋白质组学分析
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作者 李翔 张永海 +6 位作者 杨凡 张玉 杨万吉 高冉 文岳 马少玲 马汉祥 《宁夏医科大学学报》 2024年第6期571-576,共6页
目的利用蛋白质组学技术定量分析静脉注射利多卡因(IVL)前后血清中差异蛋白质的表达。方法选取健康志愿者5例,IVL 1.5 mg·kg^(-1)后再以3 mg·(kg·h)^(-1)持续泵注30 min。IVL前和IVL后1 h各采集静脉血5 mL,通过串联质谱... 目的利用蛋白质组学技术定量分析静脉注射利多卡因(IVL)前后血清中差异蛋白质的表达。方法选取健康志愿者5例,IVL 1.5 mg·kg^(-1)后再以3 mg·(kg·h)^(-1)持续泵注30 min。IVL前和IVL后1 h各采集静脉血5 mL,通过串联质谱标记及高效液相色谱分离技术鉴定IVL后差异表达蛋白,并采用京都基因和基因组百科全书(KEGG)和基因本体论(GO)数据库分析鉴定差异蛋白的生物学信息。结果检测到IVL前后的血清中差异蛋白共15种,其中上调蛋白6种、下调蛋白9种。GO分析发现,大部分差异蛋白参与了细胞进程;亚细胞结构定位发现多数差异蛋白定位于细胞外区域及细胞质;功能富集分析发现多数蛋白参与炎性反应调节、B细胞的活化调控和蛋白质加工。上调蛋白组KEGG通路富集到p53信号通路。通过对上调组和下调组差异蛋白分析,发现差异蛋白依托泊苷诱导的2.4号蛋白(EI24)参与p53信号通路且调控钙离子浓度。结论IVL可能通过调控依托泊苷诱导的EI24和上池蛋白(SBSN)抑制肿瘤细胞的发生与发展;EI24可能通过调控细胞内钙离子浓度参与IVL产生的镇静作用。 展开更多
关键词 差异蛋白 基因本体论 富集分析 串联质谱标签 利多卡因
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着丝粒蛋白U在结直肠癌患者肠组织中的表达情况及临床意义
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作者 王若淳 黄伟 +7 位作者 葛思佳 陈婧 宣晗 颜杨 姜佳炜 肖明兵 陆翠华 刘肇修 《中国现代医生》 2024年第11期1-6,共6页
目的旨在探讨着丝粒蛋白U(centromere protein U,CENPU)在结直肠癌患者肠组织中的表达情况,并结合生物信息学分析其表达水平对结直肠癌患者预后的影响。方法通过实时荧光定量聚合酶链反应(quantitative real time polymerase chain reac... 目的旨在探讨着丝粒蛋白U(centromere protein U,CENPU)在结直肠癌患者肠组织中的表达情况,并结合生物信息学分析其表达水平对结直肠癌患者预后的影响。方法通过实时荧光定量聚合酶链反应(quantitative real time polymerase chain reaction,qRT-PCR)、蛋白质免疫印迹(Western blot,WB)法以及免疫组织化学染色(immunohistochemistry,IHC)实验验证CENPU在组织中的表达情况。结合患者临床病例资料,通过单因素和多因素Cox回归分析CENPU的表达与结直肠癌患者临床病例参数的相关性;然后通过绘制受试操作者操作特征(receiver operating characteristic,ROC)曲线和Kaplan-Meier生存曲线,探究CENPU的表达对结直肠癌患者预后的预测作用。最后,通过生物信息学分析CENPU的表达对结直肠癌疾病进展影响的可能分子机制。结果通过qRT-PCR、WB法以及IHC实验均发现,与正常组织比较,CENPU在结直肠癌患者癌组织中表达显著升高。Cox回归分析表明CENPU的表达与患者的年龄和TNM分期显著相关,是影响患者预后的危险因素。Kaplan-Meier生存曲线分析表明:CENPU高表达的结直肠癌患者的生存率显著降低。ROC曲线结果表明:基于CENPU的表达建立的模型具有较高的预测结直肠癌患者预后的能力。生物信息学分析结果表明:CENPI、CENPN、CENPD、CENPK、CENPP、CENPM、CENPQ、CENPH、NDC80以及ITGB3BP这10个基因与CENPU基因具有相互作用关系;CENPU参与DNA修复、MYC/TARGETS/V1以及PI3K/AKT/MTOR等信号通路。结论结直肠癌患者癌组织中高表达的CENPU与患者的不良预后显著相关,提示CENPU有望成为结直肠癌患者早期诊断及预测预后的潜在靶点。 展开更多
关键词 结直肠癌 着丝粒蛋白U 富集分析
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O⁃连接⁃N⁃乙酰葡糖胺糖基化蛋白质的富集方法
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作者 孟领航 张威 王佳佳 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2024年第4期463-473,共11页
O-连接-N-乙酰葡糖胺(O-GlcNAc)糖基化修饰是将N-乙酰葡萄糖胺添加到细胞核和细胞质蛋白质的丝氨酸和苏氨酸残基上的一种丰富的、独特的翻译后修饰,与癌症、神经退行性病变和糖尿病等疾病密切相关。明确O-GlcNAc修饰位点是探究其在相关... O-连接-N-乙酰葡糖胺(O-GlcNAc)糖基化修饰是将N-乙酰葡萄糖胺添加到细胞核和细胞质蛋白质的丝氨酸和苏氨酸残基上的一种丰富的、独特的翻译后修饰,与癌症、神经退行性病变和糖尿病等疾病密切相关。明确O-GlcNAc修饰位点是探究其在相关疾病中调控机制的前提,同时也是临床诊断和靶向干预的关键。本文介绍了近年来O-GlcNAc糖基化修饰与疾病之间的关系以及相关修饰位点的富集方法,包括抗体和凝集素、化学酶法、亲水作用液相色谱法和非天然糖代谢标记等;此外,“凹凸理论”、“定点活化”等靶向标记特定组织内蛋白质的糖基化修饰策略已成为当前研究热点,同时基于“一石多鸟”的多功能酶法标记,以及“一锅法”分析多种糖链结构等方法也将极大促进糖蛋白组学的快速发展。总之,开发更加高效、特异的糖蛋白富集策略,将对阐明包括O-GlcNAc在内的异常糖基化修饰在相关疾病中的调控机制具有重要的研究意义。 展开更多
关键词 糖蛋白 O-连接-N-乙酰葡糖胺修饰 蛋白富集 代谢标记
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添加富含乳脂肪球膜乳清蛋白粉对发酵乳代谢组学的影响
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作者 刘菲帆 罗梦帆 +8 位作者 赵佳园 赵轩 孔维梅 王孝治 谭东虎 杨靖鹏 苏雅 罗海波 郭宇星 《中国食品学报》 EI CAS CSCD 北大核心 2024年第5期428-440,共13页
目的:研究添加富含乳脂肪球膜(MFGM)乳清蛋白粉(M-WPI)前、后的保加利亚乳杆菌发酵乳中保加利亚乳杆菌存活率,发酵乳风味及抗氧化能力。采用代谢组学分析添加M-WPI前、后保加利亚乳杆菌发酵乳的显著差异代谢标志物及其作用通路。方法:... 目的:研究添加富含乳脂肪球膜(MFGM)乳清蛋白粉(M-WPI)前、后的保加利亚乳杆菌发酵乳中保加利亚乳杆菌存活率,发酵乳风味及抗氧化能力。采用代谢组学分析添加M-WPI前、后保加利亚乳杆菌发酵乳的显著差异代谢标志物及其作用通路。方法:通过样本主成分分析、偏最小二乘判别分析、热图分析和KEGG富集分析等方法确定加入M-WPI前、后发酵乳代谢过程中的差异性代谢产物,并以此分析M-WPI对发酵乳产生的影响。结果表明:在脱脂乳中加入M-WPI,可以提高保加利亚乳杆菌存活率,增加发酵乳的风味及抗氧化活性。采用主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)共确定77种显著差异代谢物,涉及的主要代谢通路为19条。加入M-WPI后葡萄糖、半乳糖等,代谢物上调,提高了保加利亚乳杆菌存活率;癸酸、苹果酸等有机酸上调,能提高发酵乳的口感及改善发酵乳风味。保加利亚乳杆菌发酵乳中上调的代谢物以氨基酸类较多,其中丙氨酸、天冬氨酸和泛酸等参与β-丙氨酸代谢,与发酵乳抗氧化活性提高有关。本研究为加入M-WPI发酵乳营养评价以及相关加工技术和产品开发提供参考。 展开更多
关键词 富含乳脂肪球膜乳清蛋白粉 保加利亚乳杆菌 代谢组学 抗氧化 发酵乳
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基于系统生物学方法筛选癫痫潜在风险基因
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作者 郭盼 张殊悦 +1 位作者 王学韬 蔡春泉 《航天医学与医学工程》 CAS 2024年第2期91-98,共8页
目的利用生物信息学方法综合分析癫痫相关基因,从网络和通路的角度探讨其主要功能,并提取疾病关键基因。方法通过OMIM、DisGeNET、GeneCards数据库以及文献检索收集癫痫相关基因,利用“ClusterProfiler”R包进行基因本体论(GO)和通路富... 目的利用生物信息学方法综合分析癫痫相关基因,从网络和通路的角度探讨其主要功能,并提取疾病关键基因。方法通过OMIM、DisGeNET、GeneCards数据库以及文献检索收集癫痫相关基因,利用“ClusterProfiler”R包进行基因本体论(GO)和通路富集分析,通过通路串话分析识别通路之间的关联;将癫痫相关基因映射到人类蛋白质互作网络(PPIN),获得癫痫特异性蛋白互作网络,基于网络拓扑学提取疾病关键基因和潜在基因。结果收集获得572个癫痫相关基因,富集分析获得642条显著GO生物学过程条目和80条京都基因和基因组数据库(KEGG)通路,涉及学习、记忆和认知等过程;通过PPIN,提取获得10个疾病关键基因,大部分为γ–氨基丁酸(GABA)受体基因;整合PPIN和加权基因共表达网络分析(WGCNA),提取获得17个疾病潜在基因,涉及热休克蛋白、生长因子受体结合蛋白等。结论癫痫的发生是一个复杂的过程,涉及多个功能基因的异常,基因间通过紧密联系调控多个生物学过程及通路,通路间彼此交叉,共同导致疾病的发生。 展开更多
关键词 癫痫 功能富集 生化通路 蛋白质互作网络
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高尿酸血症状态下低度炎症的病理特点研究
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作者 褚梦真 王雨 +4 位作者 林志健 黄晶 刘亚迪 殷晓阳 张冰 《世界中医药》 CAS 北大核心 2024年第13期1881-1888,共8页
目的:探讨高尿酸血症(HUA)状态下低度炎症的病理特点。方法:根据体质量随机将迪法克鹌鹑分为正常组、模型组,每组10只。以普通饲料:酵母浸膏粉=4∶1制备食饵,并以该食饵喂养模型组鹌鹑,正常组鹌鹑则自由饮食饮水。分别于造模第10、20、3... 目的:探讨高尿酸血症(HUA)状态下低度炎症的病理特点。方法:根据体质量随机将迪法克鹌鹑分为正常组、模型组,每组10只。以普通饲料:酵母浸膏粉=4∶1制备食饵,并以该食饵喂养模型组鹌鹑,正常组鹌鹑则自由饮食饮水。分别于造模第10、20、30天检测血清尿酸,血清炎症介质白细胞介素-1β(IL-1β)、IL-33、IL-2、IL-13、IL-8、IL-17、IL-6、IL-10、IL-12/P40、IL-16、IL-21、C反应蛋白(CRP)、粒细胞巨噬细胞集落刺激因子(GM-CSF)、肿瘤坏死因子-α(TNF-α)、趋化因子CC配体2(CCL2)及γ干扰素(IFN-γ)、神经突起生长导向因子2(Netrin-2)、五聚蛋白3(Pentraxin 3),观察各炎症介质强度变化;造模第30天,取鹌鹑肝、回肠、肾各脏器组织,进行HE染色后观察组织病理形态变化;造模第20天,用基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析差异炎症介质功能及相关信号通路;用Pearson相关性分析方法分析差异炎症介质与血清尿酸水平的相关性。结果:与正常组比较,模型组鹌鹑血清尿酸水平高(P<0.05),以血清IL-17、IL-6、IL-33等为主的白细胞介素类,以IL-8、CCL2为主的趋化因子类,IFN-γ、TNF-α、CRP及GM-CSF水平均升高(P<0.05),而IL-13、IL-10水平降低(P<0.05)。造模第20天,GO/KEGG富集分析结果显示,HUA状态下的低度炎症可能是尿酸代谢靶点群,通过IL-17、Janus激酶信号转导和转录激活因子(JAK-STAT)等信号通路激活、细胞因子-细胞因子间相互作用,从而诱导IL-6、TNF-α等炎症介质产生。2组组织病理变化结果显示,与正常组相比,模型组回肠组织黏膜下层可见炎性细胞浸润,肝、肾组织未见明显差异。差异炎症介质与血清尿酸水平的相关性分析结果显示,鹌鹑血清中IL-6、TNF-α、CRP、IL-33、IL-17、IL-8、IFN-γ、CCL2、GM-CSF、IL-1β、IL-2、IL-6水平均与血清尿酸水平正相关,IL-10、IL-13水平与血清尿酸水平负相关。结论:HUA鹌鹑模型存在低度炎症,该低度炎症可能与尿酸代谢靶点群通过IL-17、JAK-STAT等信号通路的激活以及细胞因子间的相互作用,从而调控IL-6、TNF-α等炎症介质的产生有关。 展开更多
关键词 高尿酸血症 低度炎症 病理特点 鹌鹑 炎症介质 蛋白芯片技术 酶联免疫吸附试验法 基因本体/京都基因与基因组百科全书富集分析
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血浆蛋白与骨质疏松症的关系及潜在治疗靶点:基于国际UK Biobank数据库信息
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作者 朱凯 刘宛欣 +2 位作者 罗昊冰 冯圣一 王秋根 《中国组织工程研究》 CAS 北大核心 2025年第18期3948-3960,共13页
背景:骨质疏松症是全球范围内增加疾病负担和致残率的重要因素。血浆蛋白参与体内复杂的生物过程,在揭示疾病机制和发现潜在治疗靶点方面起着关键作用。尽管已有研究显示血浆蛋白与骨质疏松症之间存在关联,但这些关联的因果性质尚未得... 背景:骨质疏松症是全球范围内增加疾病负担和致残率的重要因素。血浆蛋白参与体内复杂的生物过程,在揭示疾病机制和发现潜在治疗靶点方面起着关键作用。尽管已有研究显示血浆蛋白与骨质疏松症之间存在关联,但这些关联的因果性质尚未得到充分阐明。因此,利用大规模的血浆蛋白数据探究与骨质疏松症相关的因果蛋白并识别潜在的药物靶点,对于骨质疏松症的防治至关重要。目的:运用两样本孟德尔随机化分析,以国际UK Biobank数据库为来源信息,评估血浆蛋白与骨质疏松症之间的因果关联。方法:从UK Biobank数据库获取1001种血浆蛋白相关的全基因组显著性水平(P<5×10-8)的蛋白质数量性状位点作为工具变量,并排除连锁不平衡。骨质疏松症的汇总数据来自FinnGen数据库,共涉及438872名欧洲血统个体。研究采用逆方差加权、MR-Egger回归、加权中位数等方法进行分析,并进行多项敏感性分析以确保结果的稳健性。进一步构建蛋白-蛋白互作网络,结合基因本体富集分析和京都基因与基因组百科全书通路分析,探索血浆蛋白的功能相关性及潜在作用机制。结果与结论:①孟德尔随机化逆方差加权法结果显示有50种血浆蛋白与骨质疏松症存在因果关系(P<0.05),包括染色体19开放阅读框12(chromosome 19 open reading frame 12,C19orf12;OR=0.610,95%CI:0.483-0.769,P=2.967×10-5)、表皮生长因子(OR=0.877,95%CI:0.770-0.999,P=0.049)等20种血浆蛋白可能与骨质疏松症的风险降低相关,有CCL18(OR=1.091,95%CI:1.037-1.147,P=0.001)、CD209(OR=1.036,95%CI:1.003-1.070,P=0.034)等30种血浆蛋白可能增加骨质疏松症的风险,经Bonferroni校正后,只有C19orf12与骨质疏松症存在显著的因果关联。②多项敏感性分析显示研究不存在多效性和异质性,说明结果具有稳健性。③通过构建蛋白-蛋白互作网络明确了表皮生长因子、CCL5、CXCL13、CXCL5、血管内皮生长因子C、CCL18、CCL17、TEK受体酪氨酸激酶、含免疫球蛋白样和表皮生长因子样结构域的酪氨酸激酶1(TIE1)和CCL23为核心蛋白。④基因本体富集分析和京都基因与基因组百科全书通路分析表明这些血浆蛋白在免疫系统中具有重要作用,通过参与信号传导、细胞迁移和趋化等过程影响骨质疏松症。⑤文章结果揭示了1001种血浆蛋白与骨质疏松症的潜在因果关联,这种基于大规模数据驱动的分析方法有助于在中国人群中识别新的生物标志物和药物靶点;其次,文章结果表明,免疫系统信号传导、细胞迁移和趋化等过程在骨质疏松症发病机制中发挥重要作用,这为特定遗传背景和环境因素下的骨质疏松症研究提供了新的方向;最后,研究中识别的核心蛋白(如表皮生长因子、CCL5及CXCL13等)有望成为新的生物标志物或药物靶点,为骨质疏松症的精准防治提供新的依据。 展开更多
关键词 血浆蛋白 蛋白质数量性状位点 骨质疏松症 孟德尔随机化 因果关系 富集分析 免疫 趋化 生物标记物 药物靶点
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