BACKGROUND As a well-known fact to the public,gestational diabetes mellitus(GDM)could bring serious risks for both pregnant women and infants.During this important investigation into the linkage between GDM patients a...BACKGROUND As a well-known fact to the public,gestational diabetes mellitus(GDM)could bring serious risks for both pregnant women and infants.During this important investigation into the linkage between GDM patients and their altered expression in the serum,proteomics techniques were deployed to detect the differentially expressed proteins(DEPs)of in the serum of GDM patients to further explore its pathogenesis,and find out possible biomarkers to forecast GDM occurrence.METHODS Subjects were divided into GDM and normal control groups according to the IADPSG diagnostic criteria.Serum samples were randomly selected from four cases in each group at 24-28 wk of gestation,and the blood samples were identified by applying iTRAQ technology combined with liquid chromatography-tandem mass spectrometry.Key proteins and signaling pathways associated with GDM were identified by bioinformatics analysis,and the expression of key proteins in serum from 12 wk to 16 wk of gestation was further verified using enzyme-linked immunosorbent assay (ELISA).RESULTS Forty-seven proteins were significantly differentially expressed by analyzing the serum samples between the GDMgravidas as well as the healthy ones. Among them, 31 proteins were found to be upregulated notably and the rest16 proteins were downregulated remarkably. Bioinformatic data report revealed abnormal expression of proteinsassociated with lipid metabolism, coagulation cascade activation, complement system and inflammatory responsein the GDM group. ELISA results showed that the contents of RBP4, as well as ANGPTL8, increased in the serumof GDM gravidas compared with the healthy ones, and this change was found to initiate from 12 wk to 16 wk ofgestation.CONCLUSION GDM symptoms may involve abnormalities in lipid metabolism, coagulation cascade activation, complementsystem and inflammatory response. RBP4 and ANGPTL8 are expected to be early predictors of GDM.展开更多
Kai-Xin-San consists of Ginseng Radix, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria at a ratio of 3:3:2:2. Kai-Xin-San has been widely used for the treatment of emotional disorders in China. However, no studi...Kai-Xin-San consists of Ginseng Radix, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria at a ratio of 3:3:2:2. Kai-Xin-San has been widely used for the treatment of emotional disorders in China. However, no studies have identified the key proteins implicated in response to Kai-Xin-San treatment. In this study, rat models of chronic mild stress were established using different stress methods over 28 days. After 14 days of stress stimulation, rats received daily intragastric administrations of 600 mg/kg Kai-Xin-San. The sucrose preference test was used to determine depression-like behavior in rats, while isobaric tags were used for relative and absolute quantitation-based proteomics to identify altered proteins following Kai-Xin-San treatment. Kai-Xin-San treatment for 2 weeks noticeably improved depression-like behaviors in rats with chronic mild stress. We identified 33 differentially expressed proteins: 7 were upregulated and 26 were downregulated. Functional analysis showed that these differentially expressed proteins participate in synaptic plasticity, neurodevelopment, and neurogenesis. Our results indicate that Kai-Xin-San has an important role in regulating the key node proteins in the synaptic signaling network, and are helpful to better understand the mechanism of the antidepressive effects of Kai-Xin-San and to provide objective theoretical support for its clinical application. The study was approved by the Ethics Committee for Animal Research from the Chinese PLA General Hospital(approval No. X5-2016-07) on March 5, 2016.展开更多
Sensory and motor nerve fibers of peripheral nerves have different anatomies and regeneration functions after injury. To gain a clear understanding of the biological processes behind these differences, we used a label...Sensory and motor nerve fibers of peripheral nerves have different anatomies and regeneration functions after injury. To gain a clear understanding of the biological processes behind these differences, we used a labeling technique termed isobaric tags for relative and absolute quantitation to investigate the protein profiles of spinal nerve tissues from Sprague-Dawley rats. In response to Wallerian degeneration, a total of 626 proteins were screened in sensory nerves, of which 368 were upregulated and 258 were downregulated. In addition, 637 proteins were screened in motor nerves, of which 372 were upregulated and 265 were downregulated. All identified proteins were analyzed using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis of bioinformatics, and the presence of several key proteins closely related to Wallerian degeneration were tested and verified using quantitative real-time polymerase chain reaction analyses. The differentially expressed proteins only identified in the sensory nerves were mainly relevant to various biological processes that included cell-cell adhesion, carbohydrate metabolic processes and cell adhesion, whereas differentially expressed proteins only identified in the motor nerves were mainly relevant to biological processes associated with the glycolytic process, cell redox homeostasis, and protein folding. In the aspect of the cellular component, the differentially expressed proteins in the sensory and motor nerves were commonly related to extracellular exosomes, the myelin sheath, and focal adhesion. According to the Kyoto Encyclopedia of Genes and Genomes, the differentially expressed proteins identified are primarily related to various types of metabolic pathways. In conclusion, the present study screened differentially expressed proteins to reveal more about the differences and similarities between sensory and motor nerves during Wallerian degeneration. The present findings could provide a reference point for a future investigation into the differences between sensory and motor nerves in Wallerian degeneration and the characteristics of peripheral nerve regeneration. The study was approved by the Ethics Committee of the Chinese PLA General Hospital, China(approval No. 2016-x9-07) in September 2016.展开更多
Objective:The objective of the study is to examine the possible mechanism by which Guasha(scraping therapy)affects the expression profiles of proteins in a lumbar disc herniation(LDH)rat model using isobaric tags for ...Objective:The objective of the study is to examine the possible mechanism by which Guasha(scraping therapy)affects the expression profiles of proteins in a lumbar disc herniation(LDH)rat model using isobaric tags for relative and absolute quantitation(iTRAQ)-based proteomics.Methods:Thirty-six rats were used in this study.LDH rats were subjected to noncompressive LDH surgeries.Rats were randomly divided into the model and Guasha groups.Guasha was applied on alternate days for a total of nine times(three courses).At the end of each course,six rats were randomly selected from each group and their blood samples were collected.iTRAQ labeling was used to examine the mechanism of action of Guasha against LDH.The molecular functions,cellular components,and biological processes were analyzed using gene ontology analysis.The Ingenuity Pathway Analysis database was used to identify canonical pathways involving these proteins.Results:Compared to the model group,198,182,and 170 proteins were identified as differentially expressed at the three respective Guasha treatment courses.Pathways,including focal adhesion kinase signaling,acute-phase response signaling,and the LXR/RXR activation pathway,were closely related to the effects of Guasha in LDH rats.Furthermore,Rac1,Orm1,and Hpx were validated by western blotting,and the results were consistent with the protein expression levels observed using the iTRAQ method.Conclusion:Guasha could not only regulate the pathological changes related to LDH,but also achieve therapeutic effects by stimulating physiological changes.Our results offer a better understanding of the effects of Guasha on LDH.展开更多
Using isobaric tags for relative and absolute quantification (iTRAQ) and associated analytic technologies, we have cataloged and compared 7 069 unique wheat proteins expressed during four substages of the filling st...Using isobaric tags for relative and absolute quantification (iTRAQ) and associated analytic technologies, we have cataloged and compared 7 069 unique wheat proteins expressed during four substages of the filling stage. Among them, 859 are differentially expressed, showing at least a 2-fold difference in concentration across substages. Differentially expressed proteins (DEPs) includind high-molecular weight giutenin subunit (W5AIU1), low-molecular weight glutenin subunit (QSW3V4), gliadin/avenin-like seed protein (D2KFG9), and avenin-like protein (W5DVL2), all of which have previously been identified as important for nutritional quality and bread-making properties, and all of which were found to increase at the latter stages of development. We have applied statistical techniques to group the proteins into hierarchical clusters, and have consulted databases to infer functional and other relationships among the identified proteins.展开更多
In treating patients with obstetric brachial plexus palsy,we noticed that denervated intrinsic muscles of the hand become irreversibly atrophic at a faster than denervated biceps.In a rat model of obstetric brachial p...In treating patients with obstetric brachial plexus palsy,we noticed that denervated intrinsic muscles of the hand become irreversibly atrophic at a faster than denervated biceps.In a rat model of obstetric brachial plexus palsy,denervated intrinsic musculature of the forepaw entered the irreversible atrophy far earlier than denervated biceps.In this study,isobaric tags for relative and absolute quantitation were examined in the intrinsic musculature of forepaw and biceps on denervated and normal sides at 3 and 5 weeks to identify dysregulated proteins.Enrichment of pathways mapped by those proteins was analyzed by Kyoto Encyclopedia of Genes and Genomes analysis.At 3 weeks,119 dysregulated proteins in denervated intrinsic musculature of the forepaw were mapped to nine pathways for muscle regulation,while 67 dysregulated proteins were mapped to three such pathways at 5 weeks.At 3 weeks,27 upregulated proteins were mapped to five pathways involving inflammation and apoptosis,while two upregulated proteins were mapped to one such pathway at 5 weeks.At 3 and 5 weeks,53 proteins from pathways involving regrowth and differentiation were downregulated.At 3 weeks,64 dysregulated proteins in denervated biceps were mapped to five pathways involving muscle regulation,while,five dysregulated proteins were mapped to three such pathways at 5 weeks.One protein mapped to inflammation and apoptotic pathways was upregulated from one pathway at 3 weeks,while three proteins were downregulated from two other pathways at 5 weeks.Four proteins mapped to regrowth and differentiation pathways were upregulated from three pathways at 3 weeks,while two proteins were downregulated in another pathway at 5 weeks.These results implicated inflammation and apoptosis as critical factors aggravating atrophy of denervated intrinsic muscles of the hand during obstetric brachial plexus palsy.All experimental procedures and protocols were approved by the Experimental Animal Ethics Committee of Fudan University,China(approval No.DF-325)in January 2015.展开更多
背景:非创伤性股骨头坏死是一类以髋关节疼痛、功能障碍进行性加重为主要症状的疾病,严重危害人体健康。它的发病机制、早期诊断和治疗仍是临床骨科医生所面临的重大挑战。目的:采用同位素标记相对和绝对定量(isobaric tags for relativ...背景:非创伤性股骨头坏死是一类以髋关节疼痛、功能障碍进行性加重为主要症状的疾病,严重危害人体健康。它的发病机制、早期诊断和治疗仍是临床骨科医生所面临的重大挑战。目的:采用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantitation,iTRAQ)技术,用蛋白质组学筛选可能与非创伤性股骨头坏死相关的关键蛋白。通过分析这些蛋白的表达特性及潜在的生物学功能,为进一步研究非创伤性股骨头坏死病因及分子机制奠定基础。方法:提取非创伤性股骨头坏死患者和健康人血清蛋白,应用iTRAQ技术标记蛋白,联合液相色谱和串联质谱分离和分析肽段。采用Proteome Discoverer 2.1软件对两组人群蛋白进行鉴定和定量分析,获得差异蛋白,再对这些表达差异蛋白进行GO注释、KEGG通路注释和功能富集聚类分析。结果与结论:①质谱鉴定共得到1006个蛋白,通过分析得到137个差异表达蛋白,其中54个蛋白表达上调,83个蛋白表达下调;②与非创伤性股骨头坏死发生相关的表达差异显著的蛋白质包括载脂蛋白B(ApoB)、载脂蛋白A2(ApoA2)、载脂蛋白E(ApoE)、补体C4和C7等;③KEGG信号通路富集分析结果显示,关键靶点的主要信号通路为p53信号通路、转录生长因子β信号通路、扩张型心肌病信号通路;④该结果提示,载脂蛋白与非创伤性股骨头坏死发生密切相关,并发现了与非创伤性股骨头坏死相关的活跃信号通路,为非创伤性股骨头坏死发生及分子机制奠定一定的实验基础。展开更多
A growing body of evidence has linked the gut microbiota to liver metabolism.The manipulation of intestinal microflora has been considered as a promising avenue to promote liver health.However,the effects of Lactobaci...A growing body of evidence has linked the gut microbiota to liver metabolism.The manipulation of intestinal microflora has been considered as a promising avenue to promote liver health.However,the effects of Lactobacillus gasseri LA39,a potential probiotic,on liver metabolism remain unclear.Accumulating studies have investigated the proteomic profile for mining the host biological events affected by microbes,and used the germ-free(GF)mouse model to evaluate host-microbe interaction.Here,we explored the effects of L.gasseri LA39 gavage on the protein expression profiles of the liver of GF mice.Our results showed that a total of 128 proteins were upregulated,whereas a total of 123 proteins were downregulated by treatment with L.gasseri LA39.Further bioinformatics analyses suggested that the primary bile acid(BA)biosynthesis pathway in the liver was activated by L.gasseri LA39.Three differentially expressed proteins(cytochrome P450 family 27 subfamily A member 1(CYP27A1),cytochrome P450 family 7 subfamily B member 1(CYP7B1),and cytochrome P450 family 8 subfamily B member 1(CYP8B1))involved in the primary BA biosynthesis pathway were further validated by western blot assay.In addition,targeted metabolomic analyses demonstrated that serum and fecalβ-muricholic acid(a primary BA),dehydrolithocholic acid(a secondary BA),and glycolithocholic acid-3-sulfate(a secondary BA)were significantly increased by L.gasseri LA39.Thus,our data revealed that L.gasseri LA39 activates the hepatic primary BA biosynthesis and promotes the intestinal secondary BA biotransformation.Based on these findings,we suggest that L.gasseri LA39 confers an important function in the gut‒liver axis through regulating BA metabolism.展开更多
目的探讨百草枯中毒后机体蛋白质表达变化及受影响的信号通路。方法采用同位素标记相对和绝对定量技术检测百草枯中毒大鼠肺组织中差异表达蛋白,对差异表达蛋白进行基因本体论(gene ontology,GO)功能富集分析和京都基因与基因组百科全书...目的探讨百草枯中毒后机体蛋白质表达变化及受影响的信号通路。方法采用同位素标记相对和绝对定量技术检测百草枯中毒大鼠肺组织中差异表达蛋白,对差异表达蛋白进行基因本体论(gene ontology,GO)功能富集分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析。结果百草枯中毒大鼠肺组织中发现84种差异蛋白,其中48种上调,36种下调。在百草枯中毒早期,差异蛋白主要存在于细胞外区域,主要涉及代谢和应激,补体和凝血级联通路、2-氧代羧酸代谢途径、神经活性配体-受体相互作用途径和金黄色葡萄球菌感染途径4条信号通路受到明显影响。结论差异蛋白和信号转导通路在百草枯致肺损伤早期即可产生变化,可能与炎症损伤和肺纤维化有关。展开更多
目的用同量异位素标记的相对和绝对定量(isobaric tags for relative and absolute quantitation,iTRAQ)技术结合液相色谱质谱技术构建3周龄小鼠与成年小鼠睾丸蛋白差异表达谱系。方法分别抽提3周龄小鼠与成年小鼠睾丸总蛋白质,iTRAQ标...目的用同量异位素标记的相对和绝对定量(isobaric tags for relative and absolute quantitation,iTRAQ)技术结合液相色谱质谱技术构建3周龄小鼠与成年小鼠睾丸蛋白差异表达谱系。方法分别抽提3周龄小鼠与成年小鼠睾丸总蛋白质,iTRAQ标记,液相色谱(LC)分离蛋白质,LTQ OrbitrapTM质谱仪进行蛋白质鉴定。结果鉴定出20个差异表达蛋白质,其中18个为3周龄小鼠睾丸高表达蛋白质,2个为成年睾丸高表达蛋白质。差异表达蛋白质主要参与细胞的有丝分裂、减数分裂、细胞增殖、分化、凋亡及精子发生等重要细胞事件。结论用目前最新的定量蛋白质组技术,构建的3周龄小鼠睾丸与成年睾丸差异表达蛋白谱系对研究睾丸功能、精子发生有一定理论价值,为定量蛋白组学技术在雄性生殖领域的应用奠定了基础。展开更多
基金This study was reviewed and approved by the Maternal and child health hospital of Hubei Province(Approval No.20201025).
文摘BACKGROUND As a well-known fact to the public,gestational diabetes mellitus(GDM)could bring serious risks for both pregnant women and infants.During this important investigation into the linkage between GDM patients and their altered expression in the serum,proteomics techniques were deployed to detect the differentially expressed proteins(DEPs)of in the serum of GDM patients to further explore its pathogenesis,and find out possible biomarkers to forecast GDM occurrence.METHODS Subjects were divided into GDM and normal control groups according to the IADPSG diagnostic criteria.Serum samples were randomly selected from four cases in each group at 24-28 wk of gestation,and the blood samples were identified by applying iTRAQ technology combined with liquid chromatography-tandem mass spectrometry.Key proteins and signaling pathways associated with GDM were identified by bioinformatics analysis,and the expression of key proteins in serum from 12 wk to 16 wk of gestation was further verified using enzyme-linked immunosorbent assay (ELISA).RESULTS Forty-seven proteins were significantly differentially expressed by analyzing the serum samples between the GDMgravidas as well as the healthy ones. Among them, 31 proteins were found to be upregulated notably and the rest16 proteins were downregulated remarkably. Bioinformatic data report revealed abnormal expression of proteinsassociated with lipid metabolism, coagulation cascade activation, complement system and inflammatory responsein the GDM group. ELISA results showed that the contents of RBP4, as well as ANGPTL8, increased in the serumof GDM gravidas compared with the healthy ones, and this change was found to initiate from 12 wk to 16 wk ofgestation.CONCLUSION GDM symptoms may involve abnormalities in lipid metabolism, coagulation cascade activation, complementsystem and inflammatory response. RBP4 and ANGPTL8 are expected to be early predictors of GDM.
基金supported by the National Natural Science Foundation of China,No.81573876(to YH)
文摘Kai-Xin-San consists of Ginseng Radix, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria at a ratio of 3:3:2:2. Kai-Xin-San has been widely used for the treatment of emotional disorders in China. However, no studies have identified the key proteins implicated in response to Kai-Xin-San treatment. In this study, rat models of chronic mild stress were established using different stress methods over 28 days. After 14 days of stress stimulation, rats received daily intragastric administrations of 600 mg/kg Kai-Xin-San. The sucrose preference test was used to determine depression-like behavior in rats, while isobaric tags were used for relative and absolute quantitation-based proteomics to identify altered proteins following Kai-Xin-San treatment. Kai-Xin-San treatment for 2 weeks noticeably improved depression-like behaviors in rats with chronic mild stress. We identified 33 differentially expressed proteins: 7 were upregulated and 26 were downregulated. Functional analysis showed that these differentially expressed proteins participate in synaptic plasticity, neurodevelopment, and neurogenesis. Our results indicate that Kai-Xin-San has an important role in regulating the key node proteins in the synaptic signaling network, and are helpful to better understand the mechanism of the antidepressive effects of Kai-Xin-San and to provide objective theoretical support for its clinical application. The study was approved by the Ethics Committee for Animal Research from the Chinese PLA General Hospital(approval No. X5-2016-07) on March 5, 2016.
基金supported by National Key Research&Development Program of China,No.2016YFC11011601,2017YFA0104701the Youth Cultivation Project of Military Medical Science,China,No.15QNP091(to YW)People’s Liberation Army Youth Training Project for Medical Science of China,No.16QNP144(to YW)
文摘Sensory and motor nerve fibers of peripheral nerves have different anatomies and regeneration functions after injury. To gain a clear understanding of the biological processes behind these differences, we used a labeling technique termed isobaric tags for relative and absolute quantitation to investigate the protein profiles of spinal nerve tissues from Sprague-Dawley rats. In response to Wallerian degeneration, a total of 626 proteins were screened in sensory nerves, of which 368 were upregulated and 258 were downregulated. In addition, 637 proteins were screened in motor nerves, of which 372 were upregulated and 265 were downregulated. All identified proteins were analyzed using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis of bioinformatics, and the presence of several key proteins closely related to Wallerian degeneration were tested and verified using quantitative real-time polymerase chain reaction analyses. The differentially expressed proteins only identified in the sensory nerves were mainly relevant to various biological processes that included cell-cell adhesion, carbohydrate metabolic processes and cell adhesion, whereas differentially expressed proteins only identified in the motor nerves were mainly relevant to biological processes associated with the glycolytic process, cell redox homeostasis, and protein folding. In the aspect of the cellular component, the differentially expressed proteins in the sensory and motor nerves were commonly related to extracellular exosomes, the myelin sheath, and focal adhesion. According to the Kyoto Encyclopedia of Genes and Genomes, the differentially expressed proteins identified are primarily related to various types of metabolic pathways. In conclusion, the present study screened differentially expressed proteins to reveal more about the differences and similarities between sensory and motor nerves during Wallerian degeneration. The present findings could provide a reference point for a future investigation into the differences between sensory and motor nerves in Wallerian degeneration and the characteristics of peripheral nerve regeneration. The study was approved by the Ethics Committee of the Chinese PLA General Hospital, China(approval No. 2016-x9-07) in September 2016.
基金the National Science Foundation of China(NSFC,No.81473791)The Third Open Subject of Nursing,Nanjing University of Chinese Medicine(No.2019YSHL028).
文摘Objective:The objective of the study is to examine the possible mechanism by which Guasha(scraping therapy)affects the expression profiles of proteins in a lumbar disc herniation(LDH)rat model using isobaric tags for relative and absolute quantitation(iTRAQ)-based proteomics.Methods:Thirty-six rats were used in this study.LDH rats were subjected to noncompressive LDH surgeries.Rats were randomly divided into the model and Guasha groups.Guasha was applied on alternate days for a total of nine times(three courses).At the end of each course,six rats were randomly selected from each group and their blood samples were collected.iTRAQ labeling was used to examine the mechanism of action of Guasha against LDH.The molecular functions,cellular components,and biological processes were analyzed using gene ontology analysis.The Ingenuity Pathway Analysis database was used to identify canonical pathways involving these proteins.Results:Compared to the model group,198,182,and 170 proteins were identified as differentially expressed at the three respective Guasha treatment courses.Pathways,including focal adhesion kinase signaling,acute-phase response signaling,and the LXR/RXR activation pathway,were closely related to the effects of Guasha in LDH rats.Furthermore,Rac1,Orm1,and Hpx were validated by western blotting,and the results were consistent with the protein expression levels observed using the iTRAQ method.Conclusion:Guasha could not only regulate the pathological changes related to LDH,but also achieve therapeutic effects by stimulating physiological changes.Our results offer a better understanding of the effects of Guasha on LDH.
基金supported by the National High-Tech R&D Program of China (863 Program,2011AA100501)the China Agricultural Research System (CARS-3-2-47)
文摘Using isobaric tags for relative and absolute quantification (iTRAQ) and associated analytic technologies, we have cataloged and compared 7 069 unique wheat proteins expressed during four substages of the filling stage. Among them, 859 are differentially expressed, showing at least a 2-fold difference in concentration across substages. Differentially expressed proteins (DEPs) includind high-molecular weight giutenin subunit (W5AIU1), low-molecular weight glutenin subunit (QSW3V4), gliadin/avenin-like seed protein (D2KFG9), and avenin-like protein (W5DVL2), all of which have previously been identified as important for nutritional quality and bread-making properties, and all of which were found to increase at the latter stages of development. We have applied statistical techniques to group the proteins into hierarchical clusters, and have consulted databases to infer functional and other relationships among the identified proteins.
基金supported by the National Natural Science Foundation of China,No.816019591003263(to JXW)the National Basic Research Program of China(973 Program),No.2014CB542203(to LC)
文摘In treating patients with obstetric brachial plexus palsy,we noticed that denervated intrinsic muscles of the hand become irreversibly atrophic at a faster than denervated biceps.In a rat model of obstetric brachial plexus palsy,denervated intrinsic musculature of the forepaw entered the irreversible atrophy far earlier than denervated biceps.In this study,isobaric tags for relative and absolute quantitation were examined in the intrinsic musculature of forepaw and biceps on denervated and normal sides at 3 and 5 weeks to identify dysregulated proteins.Enrichment of pathways mapped by those proteins was analyzed by Kyoto Encyclopedia of Genes and Genomes analysis.At 3 weeks,119 dysregulated proteins in denervated intrinsic musculature of the forepaw were mapped to nine pathways for muscle regulation,while 67 dysregulated proteins were mapped to three such pathways at 5 weeks.At 3 weeks,27 upregulated proteins were mapped to five pathways involving inflammation and apoptosis,while two upregulated proteins were mapped to one such pathway at 5 weeks.At 3 and 5 weeks,53 proteins from pathways involving regrowth and differentiation were downregulated.At 3 weeks,64 dysregulated proteins in denervated biceps were mapped to five pathways involving muscle regulation,while,five dysregulated proteins were mapped to three such pathways at 5 weeks.One protein mapped to inflammation and apoptotic pathways was upregulated from one pathway at 3 weeks,while three proteins were downregulated from two other pathways at 5 weeks.Four proteins mapped to regrowth and differentiation pathways were upregulated from three pathways at 3 weeks,while two proteins were downregulated in another pathway at 5 weeks.These results implicated inflammation and apoptosis as critical factors aggravating atrophy of denervated intrinsic muscles of the hand during obstetric brachial plexus palsy.All experimental procedures and protocols were approved by the Experimental Animal Ethics Committee of Fudan University,China(approval No.DF-325)in January 2015.
文摘背景:非创伤性股骨头坏死是一类以髋关节疼痛、功能障碍进行性加重为主要症状的疾病,严重危害人体健康。它的发病机制、早期诊断和治疗仍是临床骨科医生所面临的重大挑战。目的:采用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantitation,iTRAQ)技术,用蛋白质组学筛选可能与非创伤性股骨头坏死相关的关键蛋白。通过分析这些蛋白的表达特性及潜在的生物学功能,为进一步研究非创伤性股骨头坏死病因及分子机制奠定基础。方法:提取非创伤性股骨头坏死患者和健康人血清蛋白,应用iTRAQ技术标记蛋白,联合液相色谱和串联质谱分离和分析肽段。采用Proteome Discoverer 2.1软件对两组人群蛋白进行鉴定和定量分析,获得差异蛋白,再对这些表达差异蛋白进行GO注释、KEGG通路注释和功能富集聚类分析。结果与结论:①质谱鉴定共得到1006个蛋白,通过分析得到137个差异表达蛋白,其中54个蛋白表达上调,83个蛋白表达下调;②与非创伤性股骨头坏死发生相关的表达差异显著的蛋白质包括载脂蛋白B(ApoB)、载脂蛋白A2(ApoA2)、载脂蛋白E(ApoE)、补体C4和C7等;③KEGG信号通路富集分析结果显示,关键靶点的主要信号通路为p53信号通路、转录生长因子β信号通路、扩张型心肌病信号通路;④该结果提示,载脂蛋白与非创伤性股骨头坏死发生密切相关,并发现了与非创伤性股骨头坏死相关的活跃信号通路,为非创伤性股骨头坏死发生及分子机制奠定一定的实验基础。
基金supported by the National Natural Science Foundation of China(Nos.31925037,31730090,and 32102499)the National Postdoctoral Program for Innovative Talents of China(No.BX20190133)+1 种基金the Postdoctoral Science Foundation of China(No.2019M662671)the Natural Science Foundation of Hubei Province(Nos.2022CFB358 and 2021CFA018).
文摘A growing body of evidence has linked the gut microbiota to liver metabolism.The manipulation of intestinal microflora has been considered as a promising avenue to promote liver health.However,the effects of Lactobacillus gasseri LA39,a potential probiotic,on liver metabolism remain unclear.Accumulating studies have investigated the proteomic profile for mining the host biological events affected by microbes,and used the germ-free(GF)mouse model to evaluate host-microbe interaction.Here,we explored the effects of L.gasseri LA39 gavage on the protein expression profiles of the liver of GF mice.Our results showed that a total of 128 proteins were upregulated,whereas a total of 123 proteins were downregulated by treatment with L.gasseri LA39.Further bioinformatics analyses suggested that the primary bile acid(BA)biosynthesis pathway in the liver was activated by L.gasseri LA39.Three differentially expressed proteins(cytochrome P450 family 27 subfamily A member 1(CYP27A1),cytochrome P450 family 7 subfamily B member 1(CYP7B1),and cytochrome P450 family 8 subfamily B member 1(CYP8B1))involved in the primary BA biosynthesis pathway were further validated by western blot assay.In addition,targeted metabolomic analyses demonstrated that serum and fecalβ-muricholic acid(a primary BA),dehydrolithocholic acid(a secondary BA),and glycolithocholic acid-3-sulfate(a secondary BA)were significantly increased by L.gasseri LA39.Thus,our data revealed that L.gasseri LA39 activates the hepatic primary BA biosynthesis and promotes the intestinal secondary BA biotransformation.Based on these findings,we suggest that L.gasseri LA39 confers an important function in the gut‒liver axis through regulating BA metabolism.
文摘目的探讨百草枯中毒后机体蛋白质表达变化及受影响的信号通路。方法采用同位素标记相对和绝对定量技术检测百草枯中毒大鼠肺组织中差异表达蛋白,对差异表达蛋白进行基因本体论(gene ontology,GO)功能富集分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析。结果百草枯中毒大鼠肺组织中发现84种差异蛋白,其中48种上调,36种下调。在百草枯中毒早期,差异蛋白主要存在于细胞外区域,主要涉及代谢和应激,补体和凝血级联通路、2-氧代羧酸代谢途径、神经活性配体-受体相互作用途径和金黄色葡萄球菌感染途径4条信号通路受到明显影响。结论差异蛋白和信号转导通路在百草枯致肺损伤早期即可产生变化,可能与炎症损伤和肺纤维化有关。
文摘目的用同量异位素标记的相对和绝对定量(isobaric tags for relative and absolute quantitation,iTRAQ)技术结合液相色谱质谱技术构建3周龄小鼠与成年小鼠睾丸蛋白差异表达谱系。方法分别抽提3周龄小鼠与成年小鼠睾丸总蛋白质,iTRAQ标记,液相色谱(LC)分离蛋白质,LTQ OrbitrapTM质谱仪进行蛋白质鉴定。结果鉴定出20个差异表达蛋白质,其中18个为3周龄小鼠睾丸高表达蛋白质,2个为成年睾丸高表达蛋白质。差异表达蛋白质主要参与细胞的有丝分裂、减数分裂、细胞增殖、分化、凋亡及精子发生等重要细胞事件。结论用目前最新的定量蛋白质组技术,构建的3周龄小鼠睾丸与成年睾丸差异表达蛋白谱系对研究睾丸功能、精子发生有一定理论价值,为定量蛋白组学技术在雄性生殖领域的应用奠定了基础。