Differentially expressed proteins

差异表达蛋白
  • 文章类型: Journal Article
    多发性硬化症(MS)是中枢神经系统的脱髓鞘疾病,其特征是炎症和免疫反应增加,氧化损伤,线粒体功能障碍,铁代谢异常导致脱髓鞘和轴突损伤。在MS中,不完全的髓鞘再生导致慢性脱髓鞘的轴突和变性,同时伴有残疾。这表明MS的髓鞘再生能力失败,然而,确切的潜在机制尚不清楚.我们旨在鉴定在MS组织中慢性非活动性白质病变和斑块周围白质中表达改变的蛋白质,以揭示潜在的病理生理机制。
    激光捕获显微切割与蛋白质组学相结合,用于从三个慢性MS个体和三个没有明显神经系统并发症的对照中询问福尔马林固定石蜡包埋的脑组织的空间变化。组织病理学图指导捕获不活跃的病变,斑块周围白质,和来自慢性MS个体的皮质以及来自对照组织的相应白质和皮质。通过液相色谱串联质谱法进行的标记自由定量用于发现各个脑区域之间的差异表达的蛋白质。
    除了确认已知在MS中受影响的几种髓磷脂相关蛋白的丢失,慢性非活动性MS病变的蛋白质组学分析揭示了髓鞘组装的改变,新陈代谢,和细胞骨架组织。与对照白质相比,MS无活性病变中的最高改变蛋白由PPP1R14A组成,ERMN,SIRT2、CARNS1和MBLAC2。
    我们的发现强调了慢性非活动性MS白质病变和斑块周围白质的蛋白质组变化,这可能对正常的髓鞘形成至关重要,生物能学,粘着斑,和细胞功能。这项研究强调了空间方法的重要性和可行性,例如基于激光捕获显微切割的MS脑组织病理不同区域的蛋白质组学分析。鉴定MS脑组织蛋白质组的空间分辨变化应有助于理解病理生理机制和开发新疗法。
    UNASSIGNED: Multiple sclerosis (MS) is a demyelinating disease of the central nervous system characterized by increased inflammation and immune responses, oxidative injury, mitochondrial dysfunction, and iron dyshomeostasis leading to demyelination and axonal damage. In MS, incomplete remyelination results in chronically demyelinated axons and degeneration coinciding with disability. This suggests a failure in the ability to remyelinate in MS, however, the precise underlying mechanisms remain unclear. We aimed to identify proteins whose expression was altered in chronic inactive white matter lesions and periplaque white matter in MS tissue to reveal potential pathophysiological mechanisms.
    UNASSIGNED: Laser capture microdissection coupled to proteomics was used to interrogate spatially altered changes in formalin-fixed paraffin-embedded brain tissue from three chronic MS individuals and three controls with no apparent neurological complications. Histopathological maps guided the capture of inactive lesions, periplaque white matter, and cortex from chronic MS individuals along with corresponding white matter and cortex from control tissue. Label free quantitation by liquid chromatography tandem mass spectrometry was used to discover differentially expressed proteins between the various brain regions.
    UNASSIGNED: In addition to confirming loss of several myelin-associated proteins known to be affected in MS, proteomics analysis of chronic inactive MS lesions revealed alterations in myelin assembly, metabolism, and cytoskeletal organization. The top altered proteins in MS inactive lesions compared to control white matter consisted of PPP1R14A, ERMN, SIRT2, CARNS1, and MBLAC2.
    UNASSIGNED: Our findings highlight proteome changes in chronic inactive MS white matter lesions and periplaque white matter, which may be crucial for proper myelinogenesis, bioenergetics, focal adhesions, and cellular function. This study highlights the importance and feasibility of spatial approaches such as laser capture microdissection-based proteomics analysis of pathologically distinct regions of MS brain tissue. Identification of spatially resolved changes in the proteome of MS brain tissue should aid in the understanding of pathophysiological mechanisms and the development of novel therapies.
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  • 文章类型: Journal Article
    本研究旨在利用Olink蛋白质组学分析与绝经后骨质疏松症(PMOP)相关的不同血清蛋白,并通过绝经后骨质疏松症的分子机制研究,确定早期检测PMOP的预后标志物。选取北京积水潭医院收治的绝经后妇女,根据双能X线骨密度仪(DXA)T评分分为3组:骨质疏松组(n=24),骨量减少组(n=20),正常骨量组(n=16)。收集来自所有参与者的血清样品用于临床和骨代谢标志物测量。Olink蛋白质组学用于鉴定与绝经后骨质疏松症高度相关的差异表达蛋白(DEP)。使用基因本体论和Kyto百科全书基因和基因组(KEGG)进行DEP的功能分析。随后分析了这些蛋白质的生物学特性及其与PMOP的相关性。进行ROC曲线分析以鉴定对早期PMOP具有最高诊断准确性的潜在生物标志物。通过Olink蛋白质组学,我们确定了五个与PMOP高度相关的DEP,包括两个上调和三个下调的蛋白质。TWEAK和CDCP1标志物表现出最高的曲线下面积(分别为0.8188和0.8031)。TWEAK和CDCP1有可能作为早期预测绝经后骨质疏松症的生物标志物。
    This investigation aims to employ Olink proteomics in analyzing the distinct serum proteins associated with postmenopausal osteoporosis (PMOP) and identifying prognostic markers for early detection of PMOP via molecular mechanism research on postmenopausal osteoporosis. Postmenopausal women admitted to Beijing Jishuitan Hospital were randomly selected and categorized into three groups based on their dual-energy X-ray absorptiometry (DXA) T-scores: osteoporosis group (n = 24), osteopenia group (n = 20), and normal bone mass group (n = 16). Serum samples from all participants were collected for clinical and bone metabolism marker measurements. Olink proteomics was utilized to identify differentially expressed proteins (DEPs) that are highly associated with postmenopausal osteoporosis. The functional analysis of DEPs was performed using Gene Ontology and Kyto Encyclopedia Genes and Genomes (KEGG). The biological characteristics of these proteins and their correlation with PMOP were subsequently analyzed. ROC curve analysis was performed to identify potential biomarkers with the highest diagnostic accuracy for early stage PMOP. Through Olink proteomics, we identified five DEPs highly associated with PMOP, including two upregulated and three downregulated proteins. TWEAK and CDCP1 markers exhibited the highest area under the curve (0.8188 and 0.8031, respectively). TWEAK and CDCP1 have the potential to serve as biomarkers for early prediction of postmenopausal osteoporosis.
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  • 文章类型: Journal Article
    锌(Zn)缺乏是全球作物产量的显著营养限制,特别是在钙质土壤环境中。牡丹的牡丹因其种子富含α-亚麻酸而被视为油料作物,一种促进健康的有益化合物。然而,低种子产量仍然是从牡丹中获得足够种子油的主要挑战。在这项研究中,在果实发育前的生长期,对虎尾草的土壤或叶子进行了锌施肥。我们的发现表明,叶面喷锌,与土壤施用相反,被证明是增加种子产量的更有效方法,凤丹锌的积累和光合能力。在叶面喷锌下使用iTRAQ分析对“凤丹”的叶片蛋白质组进行比较分析,鉴定出115种差异表达蛋白(DEP),包括36种上调的蛋白质,这可能有助于观察到由叶面喷施锌引起的“凤丹”种子产量的增加。具体来说,磷脂酰肌醇信号传导的Zn2+刺激引发代谢调节的级联。首先,ATP合成促进叶片光合能力,通过上调支链淀粉酶和1,4-α-葡聚糖分支酶改善蔗糖代谢而促进。此外,脂质合成和转运是由上调的脂酰合酶和质体脂质相关蛋白促进的。此外,参与次生代谢的DEP在各种有利于“凤丹”生长的代谢物的产生中被上调。总的来说,我们的结果表明,叶面喷施锌可通过提高叶中锌含量和次生代谢产物的合成来提高紫藤的种子产量。从而增强叶片光合能力和脂质合成。本研究为外源施锌提高牡丹种子产量提供了有效途径。
    Zinc (Zn) deficiency is a significant nutritional limitation to crop yield globally, particularly in calcareous soil environments. Tree peony of Peaonia ostii \'Fengdan\' is regarded as an oil crop due to its seeds rich in alpha-linolenic acid, a beneficial compound for health promotion. However, low seed yield remains a primary challenge in attaining sufficient seed oil from tree peony. In this study, Zn fertilization was applied to soil or foliage of P. ostii \'Fengdan\' in the growth period before fruit development. Our findings reveal that foliar Zn-spraying, as opposed to soil application, proves to be a more effective method for augmenting seed yield, Zn accumulation and photosynthetic capacity in \'Fengdan\'. Comparative analyses of the leaf proteome of \'Fengdan\' using iTRAQ profiling under foliar Zn-spraying identified 115 differentially expressed proteins (DEPs), including 36 upregulated proteins, which likely contribute to the observed increase in seed yields of \'Fengdan\' caused by foliage Zn-spraying. Specifically, Zn2+ stimulation of phosphatidylinositol signaling initiates a cascade of metabolic regulations. Firstly, ATP synthesis promotes leaf photosynthetic capacity, facilitated by improved sucrose metabolism through upregulated pullulanase and 1,4-alpha-glucan-branching enzyme. Furthermore, lipid synthesis and transport are facilitated by upregulated lipoyl synthase and plastid lipid-associated proteins. Additionally, DEPs involved in secondary metabolism are upregulated in the production of various metabolites conducive to \'Fengdan\' growth. Overall, our results demonstrate that foliage Zn-spraying enhances seed yield in P. ostii \'Fengdan\' by elevating Zn content and secondary metabolite synthesis in leaves, thereby augmenting leaf photosynthetic capacity and lipid synthesis. This study provides an effective way to increase seed yield of tree peony by exogenous Zn application.
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  • 文章类型: Journal Article
    热应激是水产养殖物种的主要问题,导致生理变化,如减少采食量,增长率,繁殖,和内部细胞损伤,从而影响鱼类的健康。在本研究中,在罗胡(Labeorohita)鱼种的肝脏蛋白质组中评估了模拟夏季温度每日上升和下降的急性热应激的影响。将保持在30°C的鱼逐渐暴露于36°C的较高温度下,每1.5小时递增速率为1°C,在该温度下3小时后,逐渐降低到30℃。在凌晨5点收集肝组织样本,下午5点,第二天凌晨5点,从暴露和控制的鱼开始。从肝脏组织制备蛋白质样品,和提取的蛋白质使用2维(2D)凝胶电泳(2DGE)和质谱(MS)使用MALDI-TOF/TOF质谱仪进行比较。在三个时间点,从热应激暴露鱼的2D凝胶分析中观察到总共44个差异表达的蛋白质点,其中包括一种假设蛋白质的21种蛋白质可以被MS鉴定。使用qPCR验证五种选择的差异表达的蛋白质(DEP)的丰度。发现大多数DEP主要与脂质有关,蛋白质和能量代谢,免疫系统调节,细胞骨架稳定性,和ROS管理。这项研究的结果将有助于制定减轻L.rohita热应激的策略。
    Heat stress is a major problem in aquaculture species, causing changes in physiology such as decreased feed intake, growth rate, reproduction, and internal cellular damage, thereby affecting fish\'s health. The effects of an acute heat stress simulating a daily rise and fall in temperature on summer days were evaluated in the liver proteome of rohu (Labeo rohita) fingerlings in the present study. The fish maintained at 30 °C were gradually exposed to a higher temperature of 36 °C at an increment rate of 1 °C per 1.5 h, and after 3 h at that temperature, it was gradually reduced to 30 °C. The liver tissue samples were collected at 5 am, 5 pm, and 5 am the next day from the exposed and control fish. Protein samples were prepared from the liver tissues, and the extracted proteins were compared using 2-dimensional (2D) gel electrophoresis (2DGE) and mass spectrometry (MS) using a MALDI-TOF/TOF mass spectrometer. A total of 44 differentially expressed protein spots were visualized in 2D gel analysis from heat stress exposed fish at three time points, out of which 21 proteins including one hypothetical protein could be identified by MS. The abundance of five selected differentially expressed proteins (DEPs) was validated using qPCR. The majority of DEPs were found to be involved primarily in lipid, protein and energy metabolism, immune system regulation, cytoskeletal stability, and ROS management. The findings of this study would help in the development of strategies to mitigate heat stress in L. rohita.
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  • 文章类型: Journal Article
    弥漫性肺泡出血(DAH)是一种灾难性的临床综合征,是系统性红斑狼疮(SLE)肺部受累的表现之一,以咯血为特征,弥漫性肺浸润,和呼吸衰竭。然而,DAH的治疗选择仍然有限,与DAH相关的研究需要探索更有效的治疗方向,以改善疾病管理和预后.
    本研究利用了前列腺素诱导的DAH小鼠模型来模拟SLE患者DAH的病理过程。进行蛋白质组学分析以检测存活和非存活小鼠血浆中的差异表达蛋白(DEP),其次是生物学功能和途径的分析。然后在有或没有DAH和有或没有致命结局的DAH鼠模型的SLE患者的血浆中证实了最显著的DEP。最后,结合珠蛋白(Hp)替代的治疗价值在DAH小鼠模型中通过肺组织病理学,RT-qPCR,和生存分析。
    这项研究确定了178个DEP,在非存活组中有118个上调和60个下调的DEP。在一组著名的京都基因和基因组百科全书(KEGG)途径中,补体和凝血级联是与DAH过程相关的最突出的途径。稍后,最重要的DEP,触珠蛋白(Hp),通过ELISA测试证实,SLE-DAH和DAH小鼠模型的血浆显着降低,结果较差。最后,与对照组相比,Hp治疗组DAH的严重程度明显减轻,表现为促炎细胞因子(IL-6和TNF-α)水平降低,抗炎细胞因子(IL-10和TGF-β)水平升高,降低死亡率。
    在SLE-DAH中观察到血浆Hp水平降低,Hp替代疗法可以减轻DAH小鼠的肺出血并降低死亡率。这项研究确定了Hp作为其临床诊断和治疗方向的潜在生物标志物。
    UNASSIGNED: Diffuse alveolar hemorrhage (DAH) is a catastrophic clinical syndrome and one of the manifestations of pulmonary involvement in systemic lupus erythematosus (SLE), which is characterized by hemoptysis, diffuse pulmonary infiltrates, and respiratory failure. However, the treatment options for DAH remain limited, and DAH-related studies are needed to explore more effective therapeutic directions for better disease management and improved prognosis.
    UNASSIGNED: This study utilized the pristane-induced DAH murine model to mimic the pathological process of DAH in patients with SLE. Proteomic analysis was conducted to detect differentially expressed proteins (DEPs) in the plasma of surviving and non-surviving mice, followed by an analysis of biological functions and pathways. The most significant DEP was then confirmed in the plasma of SLE patients with or without DAH and DAH murine model with or without fatal outcomes. Finally, the therapeutic value of haptoglobin (Hp) replacement was validated in a DAH murine model through lung histopathology, RT-qPCR, and survival analysis.
    UNASSIGNED: This study identified 178 DEPs, with 118 upregulated and 60 downregulated DEPs in the non-survival group. Within a set of notable Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, complement and coagulation cascades emerged as the most prominent pathway associated with the process of DAH. Later, the most significant DEP, haptoglobin (Hp), was confirmed to exhibit a significant decrease in the plasma of individuals with SLE-DAH and DAH murine model with poor outcomes by the ELISA test. Finally, compared with the control group, the severity of DAH in the Hp treatment group was alleviated significantly, as manifested by the decreased levels of pro-inflammatory cytokines (IL-6 and TNF-α), increased levels of anti-inflammatory cytokines (IL-10 and TGF-β), and decreased mortality.
    UNASSIGNED: A reduction in plasma Hp levels was observed in SLE-DAH, and the replacement therapy with Hp could alleviate pulmonary hemorrhage and reduce mortality in DAH mice. This study identified Hp as a potential biomarker for its clinical diagnosis and a direction for treatment.
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  • 文章类型: Journal Article
    神经突形成的分子机制包括途径之间的多种串扰,例如膜运输,细胞内信号,和肌动蛋白细胞骨架重排。为了研究参与这种复杂途径的蛋白质,我们介绍了基于质谱的蛋白质组学和数据分析的样品制备的详细工作流程.我们还包括对蛋白质进行无标记定量的步骤,这将有助于研究人员在全球范围内量化神经元形态发生关键调节因子的表达水平的变化。
    The molecular mechanisms underlying neurite formation include multiple crosstalk between pathways such as membrane trafficking, intracellular signaling, and actin cytoskeletal rearrangement. To study the proteins involved in such complex pathways, we present a detailed workflow of the sample preparation for mass spectrometry-based proteomics and data analysis. We have also included steps to perform label-free quantification of proteins that will help researchers quantify changes in the expression levels of key regulators of neuronal morphogenesis on a global scale.
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  • 文章类型: Journal Article
    背景:差异表达基因/蛋白质(DEG/DEP)在肺结核(PTB)的诊断和治疗中起着至关重要的作用。然而,缺乏关于PTB患者肺组织和血液样本中DEG/DEP的报道。
    目的:我们旨在鉴定PTB患者肺组织和血液样本中的DEG/DEP,并研究其在PTB中的作用。
    方法:从PTB患者收集肺肉芽肿和正常组织用于蛋白质组学和转录组学分析。基因本体论(GO)和京都基因和基因组百科全书(KEGG)分析注释了DEG/DEP的功能。下载GSE107994数据集以鉴定外周血中的DEG/DEP。确定了常见的DEG和DEP。建立了一个列线图。进行Pearson相关分析。
    结果:确定了83个DEG/DEP。这些DEG/DEP主要富集在细胞或亚细胞组分的运动中,细胞成分生物发生的调节,和基于肌动蛋白丝的过程以及肌醇磷酸代谢的途径,附着者接合处,磷脂酰肌醇信号系统,白细胞跨内皮迁移,肌动蛋白细胞骨架的调节,和紧密连接。共有八个DEG/DEP(TYMP,LAP3,ADGRL2,SIL1,LMO7,SULF1,ANXA3和PACSIN3)在肺组织和血液样品之间。它们在预测结核病方面是有效的。此外,激活的树突状细胞,巨噬细胞,单核细胞,中性粒细胞,调节性T细胞与TYMP呈显著正相关(r>.50),LAP3(r>.50),SIL1(r>.50),ANXA3(r>.5),和PACSIN3(r<.50),而与LMO7呈负相关(r<-0.50)(p<0.05)。ADGRL2与SULF1无显著相关性(p>.05)。
    结论:样本量小。
    结论:确定了8种常见的肺组织和血液样本的DEG/DEP。它们与免疫细胞相关,并证明了对PTB的预测价值。我们的数据可能有助于PTB的诊断和治疗。
    BACKGROUND: Differentially expressed genes/proteins (DEGs/DEPs) play critical roles in pulmonary tuberculosis (PTB) diagnosis and treatment. However, there is a scarcity of reports on DEGs/DEPs in lung tissues and blood samples in PTB patients.
    OBJECTIVE: We aim to identify the DEGs/DEPs in lung tissues and blood samples of PTB patients and investigate their roles in PTB.
    METHODS: The lung granulomas and normal tissues were collected from PTB patients for proteomic and transcriptomic analyses. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses annotated the functions of DEGs/DEPs. The GSE107994 data set was downloaded to identify the DEGs/DEPs in peripheral blood. The common DEGs and DEPs were identified. A nomogram was established. Pearson correlation analysis was conducted.
    RESULTS: Eighty-three DEGs/DEPs were identified. These DEGs/DEPs were mainly enriched in the movement of cell or subcellular components, regulation of cellular component biogenesis, and actin filament-based process as well as in the pathways of inositol phosphate metabolism, adherens junction, phosphatidylinositol signaling system, leukocyte transendothelial migration, regulation of actin cytoskeleton, and tight junction. There were eight common DEGs/DEPs (TYMP, LAP3, ADGRL2, SIL1, LMO7, SULF 1, ANXA3, and PACSIN3) between the lung tissues and blood samples. They were effective in predicting tuberculosis. Moreover, the activated dendritic cells, macrophages, monocytes, neutrophils, and regulatory T cells were significantly positively correlated with TYMP (r > .50), LAP3 (r > .50), SIL1 (r > .50), ANXA3 (r > .5), and PACSIN3 (r < .50), while negatively correlated with LMO7 (r < -0.50) (p < .05). ADGRL2 and SULF1 did not have a significant correlation (p > .05).
    CONCLUSIONS: The sample size was small.
    CONCLUSIONS: Eight common DEGs/DEPs of lung tissues and blood samples were identified. They were correlated with immune cells and demonstrated predictive value for PTB. Our data may facilitate the diagnosis and treatment of PTB.
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  • 文章类型: Journal Article
    骨骼肌生长是肉类生产的重要经济性状,藏猪之间存在显著差异(TIBPs,生长缓慢的品种)和大型白猪(LWP,快速增长的品种)。然而,这种差异的遗传基础仍不清楚.
    在当前的研究中,我们整合了来自60日龄胚胎肌肉组织的差异表达基因(DEG)和蛋白质(DEP),随着全基因组单核苷酸多态性(SNP)显示绝对等位基因频率差异(ΔAF)在TIBP和LWP品种之间0.5或更多,解开影响骨骼肌生长的遗传因素。
    我们的分析揭示了3499DEG和628DEP,其中SNP的ΔAF等于或大于0.5。进一步的功能分析确定了145个DEGs和23个DEP参与与骨骼肌发育相关的生物过程,22个DEGs和3个DEP参与mTOR信号通路,以积极调节蛋白质合成而闻名。在这些基因中,几个DEG和DEP,在调控区或/和编码区富含TIPB特异性SNP,在TIBP和LWP品种之间显示出明显的ΔAF,包括MYF5,MYOF,ASB2,PDE9A,SDC1,PDGFRA,MYOM2,ACVR1,ZIC3,COL11A1,TGFBR1,EDNRA,TGFB2,PDE4D,PGAM2,GRK2,SCN4B,CACNA1S,MYL4、IGF1和FOXO1。此外,CAPN3,MYOM2和PGAM2等基因被鉴定为与骨骼肌发育相关的DEP和DEG,在调控区和/或编码区包含多个TIBP特异性和LWP优势SNP,强调了两个品种之间显著的ΔAF差异。
    s:这项对DEG和DEP中SNP的全面调查发现了大量与产前阶段骨骼肌发育相关的SNP和基因。这些发现不仅揭示了TIBP和LWP品种之间肌肉差异的潜在因果基因,而且为旨在提高肉类产量的猪育种策略提供了有价值的见解。
    UNASSIGNED: Skeletal muscle growth is an important economic trait for meat production, with notable differences between Tibetan pigs (TIBPs, a slow-growing breed) and Large White pigs (LWPs, a fast-growing breed). However, the genetic underpinnings of this disparity remain unclear.
    UNASSIGNED: In the current study, we integrated differentially expressed genes (DEGs) and proteins (DEPs) from 60-day-old embryonic muscle tissue, along with whole-genome single nucleotide polymorphisms (SNPs) displaying absolute allele frequency differences (ΔAF) of 0.5 or more between the TIBP and LWP breeds, to unravel the genetic factors influencing skeletal muscle growth.
    UNASSIGNED: Our analysis revealed 3499 DEGs and 628 DEPs with SNPs having a ΔAF equal to or greater than 0.5. Further functional analysis identified 145 DEGs and 23 DEPs involved in biological processes related to skeletal muscle development, and 22 DEGs and 3 DEPs implicated in the mTOR signaling pathway, which is known for positively regulating protein synthesis. Among these genes, several DEGs and DEPs, enriched with TIPB-specific SNPs in regulatory or/and coding regions, showed marked ΔAF between the TIBP and LWP breeds, including MYF5, MYOF, ASB2, PDE9A, SDC1, PDGFRA, MYOM2, ACVR1, ZIC3, COL11A1, TGFBR1, EDNRA, TGFB2, PDE4D, PGAM2, GRK2, SCN4B, CACNA1S, MYL4, IGF1, and FOXO1. Additionally, genes such as CAPN3, MYOM2, and PGAM2, identified as both DEPs and DEGs related to skeletal muscle development, contained multiple TIBP-specific and LWP-predominant SNPs in regulatory and/or coding regions, underscoring significant ΔAF differences between the two breeds.
    UNASSIGNED: s: This comprehensive investigation of SNPs in DEGs and DEPs identified a significant number of SNPs and genes related to skeletal muscle development during the prenatal stage. These findings not only shed light on potential causal genes for muscle divergence between the TIBP and LWP breeds but also offer valuable insights for pig breeding strategies aimed at enhancing meat production.
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  • 文章类型: Journal Article
    WT9-12是常用于常染色体显性多囊肾病(ADPKD)研究的细胞系之一。先前的研究已经描述了WT9-12中的PKD基因突变和多囊素表达。尽管如此,其他ADPKD相关基因中发生的突变尚未被研究.这项研究旨在重新审视WT9-12的这些突变和蛋白质谱。全基因组测序证实在WT9-12的PKD1基因中在氨基酸2556(Q2556X)处存在截短突变。此外,影响较大的变异包括PKD1L2中的单核苷酸多态性(rs8054182,rs117006360和rs12925771)和插入和缺失(InDels)(rs145602984和rs55980345);PKD1L3中的InDel(rs1296698195);以及GANAB中的拷贝数变异.使用等量异位标签比较从WT9-12和HK-2细胞的总蛋白产生的蛋白谱,用于相对和绝对定量(iTRAQ)分析。多囊藻毒素-1在WT9-12中不存在。基因本体论富集和反应小体途径分析显示,WT9-12相对于HK-2细胞系的上调和下调蛋白导致与免疫应答和氨基酸代谢相关的信号通路,分别。与WT9-12中差异表达蛋白相关的ADPKD相关突变和信号通路可能有助于研究人员选择细胞系进行研究。
    WT 9-12 is one of the cell lines commonly used for autosomal dominant polycystic kidney disease (ADPKD) studies. Previous studies had described the PKD gene mutations and polycystin expression in WT 9-12. Nonetheless, the mutations occurring in other ADPKD-associated genes have not been investigated. This study aims to revisit these mutations and protein profile of WT 9-12. Whole genome sequencing verified the presence of truncation mutation at amino acid 2556 (Q2556X) in PKD1 gene of WT 9-12. Besides, those variations with high impacts included single nucleotide polymorphisms (rs8054182, rs117006360, and rs12925771) and insertions and deletions (InDels) (rs145602984 and rs55980345) in PKD1L2; InDel (rs1296698195) in PKD1L3; and copy number variations in GANAB. Protein profiles generated from the total proteins of WT 9-12 and HK-2 cells were compared using isobaric tags for relative and absolute quantitation (iTRAQ) analysis. Polycystin-1 was absent in WT 9-12. The gene ontology enrichment and reactome pathway analyses revealed that the upregulated and downregulated proteins of WT 9-12 relative to HK-2 cell line leaded to signaling pathways related to immune response and amino acid metabolism, respectively. The ADPKD-related mutations and signaling pathways associated with differentially expressed proteins in WT 9-12 may help researchers in cell line selection for their studies.
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  • 文章类型: Journal Article
    COVID-19是一种高度传染性的呼吸道疾病,其进展与多种因素有关。从SARS-CoV-2感染到死亡,需要能够预测不同疾病过程的生物标志物来帮助我们进一步了解COVID-19疾病的分子进展。目的是寻找与COVID-19疾病进展相关或可能是潜在生物标志物的差异表达蛋白,为进一步理解COVID-19发生的分子机制提供参考,programming,和治疗。数据独立采集(DIA)蛋白质组学获得样本蛋白质表达数据,使用R语言筛选差异表达的蛋白质。对差异蛋白质进行基因本体论和京都百科全书的基因和基因组分析,并构建蛋白质-蛋白质相互作用(PPI)网络以筛选关键蛋白质。共从COVID-19孵育患者和健康人群中获得47种差异表达蛋白(L/H),主要富含血小板相关功能,补体和凝血级联反应途径,如血小板脱颗粒和血小板聚集。在临床和潜伏期患者(C/L)中共获得42种差异蛋白,还主要富含血小板相关功能以及补体和凝血级联反应,血小板活化途径。在恢复期和临床期患者(R/C)共筛选了10种差异蛋白,主要是免疫相关的蛋白质。COVID-19不同阶段的差异表达蛋白大多与凝血密切相关,和关键的差异蛋白质,比如FGA,FGB,FGG,ACTB,PFN1,VCL,SERPZNCL,APOC3,LTF,和DEFA1,有潜力作为早期诊断标志物。
    COVID-19 is a highly infectious respiratory disease whose progression has been associated with multiple factors. From SARS-CoV-2 infection to death, biomarkers capable of predicting different disease processes are needed to help us further understand the molecular progression of COVID-19 disease. The aim is to find differentially expressed proteins that are associated with the progression of COVID-19 disease or can be potential biomarkers, and to provide a reference for further understanding of the molecular mechanisms of COVID-19 occurrence, progression, and treatment. Data-independent Acquisition (DIA) proteomics to obtain sample protein expression data, using R language screening differentially expressed proteins. Gene Ontology and Kyoto Encyclopedia for Genes and Genomes analysis was performed on differential proteins and protein-protein interaction (PPI) network was constructed to screen key proteins. A total of 47 differentially expressed proteins were obtained from COVID-19 incubation patients and healthy population (L/H), mainly enriched in platelet-related functions, and complement and coagulation cascade reaction pathways, such as platelet degranulation and platelet aggregation. A total of 42 differential proteins were obtained in clinical and latent phase patients (C/L), also mainly enriched in platelet-related functions and in complement and coagulation cascade reactions, platelet activation pathways. A total of 10 differential proteins were screened in recovery and clinical phase patients (R/C), mostly immune-related proteins. The differentially expressed proteins in different stages of COVID-19 are mostly closely associated with coagulation, and key differential proteins, such as FGA, FGB, FGG, ACTB, PFN1, VCL, SERPZNCL, APOC3, LTF, and DEFA1, have the potential to be used as early diagnostic markers.
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