目的筛选腰椎间盘突出症两种不同非手术治疗前后患者血清差异表达蛋白,进一步明确LIDP活血化瘀治疗的血清标志蛋白。方法纳入60例确诊为LIDP并中医辨证为血瘀证的患者,随机分为观察组和对照组,每组30例,观察组给予中医活血化瘀法治疗,...目的筛选腰椎间盘突出症两种不同非手术治疗前后患者血清差异表达蛋白,进一步明确LIDP活血化瘀治疗的血清标志蛋白。方法纳入60例确诊为LIDP并中医辨证为血瘀证的患者,随机分为观察组和对照组,每组30例,观察组给予中医活血化瘀法治疗,对照组用西医非手术综合治疗。基于i TRAQ联合LC-MS/MS分析和鉴定两组患者治疗前后的血清差异表达蛋白质,采用Gene Ontology及KEGG对相关生物信息、差异蛋白的功能和代谢通路进行分析,同时记录治疗前后两组患者VAS及JOA评分,并注意观测是否出现不良反应。结果 i TRAQ联合LC-MS/MS质谱共鉴定到蛋白质组为300个,对应209个非冗余蛋白,观察组符合条件的差异蛋白20个,其中在LIDP血瘀证患者中呈上调表达及下调表达的差异蛋白各占10个,9个差异蛋白序列表达涉及27条KEGG生物代谢途径;对照组符合条件的差异蛋白23个,其中呈上调表达的差异蛋白13个,下调10个,14个差异蛋白序列表达涉及25条KEGG生物代谢途径;筛选出两组共同差异蛋白8个,其中呈上调表达的差异蛋白3个,下调5个,两组共同有6个差异蛋白富集到19条信号通路上,有2个差异蛋白为观察组所特有。生物学信息分析结果显示,代表性差异表达蛋白碳酸酐酶1与转铁蛋白变体在观察组LIDP患者血清中表达量下调,差异具有统计学意义(P<0.05)。两组患者治疗后较治疗前的VAS评分均明显降低,JOA评分明显升高(P<0.05),但两组VAS、JOA评分及总体疗效比较无显著差异(P>0.05)。结论采用活血化瘀治疗血瘀证LIDP可取得与西医非手术综合疗法几乎相同的疗效,治疗是对证的;碳酸酐酶1与转铁蛋白变体是活血化瘀法治疗腰椎间盘突出症血瘀证的目标蛋白质。展开更多
橡胶树死皮(Tapping panel dryness,TPD)是限制天然橡胶产量提高的关键因子之一,鉴定健康和死皮橡胶树胶乳差异表达蛋白对阐明橡胶树死皮发生机制具有重要意义。研究首次采用同位素标记相对和绝对定量(Isobaric tags for relative and a...橡胶树死皮(Tapping panel dryness,TPD)是限制天然橡胶产量提高的关键因子之一,鉴定健康和死皮橡胶树胶乳差异表达蛋白对阐明橡胶树死皮发生机制具有重要意义。研究首次采用同位素标记相对和绝对定量(Isobaric tags for relative and absolute quantitation,iTRAQ)结合二维液相色谱串联质谱(Two-dimensional liquid chromatography-tandem mass spectrometry,2D LC-MS/MS)技术对健康和死皮橡胶树胶乳蛋白质组的差异进行研究。结果显示,有16个蛋白差异表达,其中,在死皮树中表达上调的蛋白有2个,下调的蛋白有14个。进一步应用实时荧光定量RT-PCR对其中6个差异蛋白的基因进行表达分析,发现其表达趋势与蛋白质组水平的研究结果一致。功能分析显示,这些差异蛋白主要与细胞程序性死亡(如TCTP、HSP70、CaM、cysteine protease等)和橡胶生物合成(如SRPP和diphosphomevelonate decarboxylase)相关,从蛋白质组学角度进一步验证了橡胶树死皮发生过程中涉及细胞程序性死亡和橡胶生物合成途径。展开更多
首次利用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)技术结合二维液相色谱串联质谱对草菇(Volvariella volvacea)同核和异核菌丝蛋白质组进行研究。采用QExactive质谱鉴定并经MASCOT软...首次利用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)技术结合二维液相色谱串联质谱对草菇(Volvariella volvacea)同核和异核菌丝蛋白质组进行研究。采用QExactive质谱鉴定并经MASCOT软件搜库,对所有鉴定蛋白进行主成分分析(principal component analysis,PCA)、层次聚类(hierarchical clustering)分析和GO(gene ontology)注释,并进行生物信息学分析。结果共获得2335个不同肽段,鉴定到1039个蛋白,其中1030个蛋白具有定量信息。在同核菌丝中显著上调蛋白83个,下调蛋白97个,表明iTRAQ标记技术结合二维液相色谱串联质谱可有效地分离和鉴定草菇菌丝蛋白质组。该研究结果为更深入全面地研究草菇菌丝蛋白质组学奠定基础。展开更多
目的分析家蝇幼虫免疫诱导前后血淋巴中的免疫防御相关靶标,探索其先天性免疫机制。方法采用同位素标记及相对定量技术(isobaric tags for relative and absolute quantification,iTRAQ)结合2D LC-MS/MS对家蝇幼虫诱导前后的血淋巴蛋白...目的分析家蝇幼虫免疫诱导前后血淋巴中的免疫防御相关靶标,探索其先天性免疫机制。方法采用同位素标记及相对定量技术(isobaric tags for relative and absolute quantification,iTRAQ)结合2D LC-MS/MS对家蝇幼虫诱导前后的血淋巴蛋白质组进行研究。结果与对照组比较研究后共获得237个不同肽段,鉴定到13个具有定量信息的差异蛋白。免疫刺激后显著上调的蛋白有9个,下调蛋白4个(P<0.05),对鉴定到的差异蛋白进行理化性质分析和GO(gene ontology)注释分析,发现这些蛋白分别具有抗菌、抗氧化、催化结合等功能,并参与了免疫、代谢、应激、转运等生物学过程,表明对家蝇幼虫经诱导后血淋巴中多种功能蛋白的表达发生了变化。结论 iTRAQ标记技术结合2D LC-MS/MS可以有效地分离鉴定昆虫血淋巴蛋白质组,为深入研究差异蛋白的功能奠定了基础。展开更多
目的:筛选胃癌血清差异表达蛋白,寻找胃癌血清标志物.方法:采集胃癌患者血清和正常对照人群血清各45例,所有血清样本去除高丰度血清蛋白质后,采用相对和绝对定量同位素标志(isobaric tags for relative and absolute q u a n t i t a t ...目的:筛选胃癌血清差异表达蛋白,寻找胃癌血清标志物.方法:采集胃癌患者血清和正常对照人群血清各45例,所有血清样本去除高丰度血清蛋白质后,采用相对和绝对定量同位素标志(isobaric tags for relative and absolute q u a n t i t a t i o n,i T R A Q)技术联合二维液相色谱-串联质谱(two-dimensional liquid chromatography/tandem mass spectrometry,2D-LC-MS/MS)分析和鉴定两组间的差异表达蛋白;采用生物信息学对差异蛋白进行分析;用Western blot对蛋白组中的代表性血清差异表达蛋白进行表达验证.结果:iTRAQ标记联合2D-LC-MS/MS分析一共鉴定出10540条独特肽段,对应199个非冗余蛋白,其中符合条件的差异蛋白17个,其中12个蛋白在胃癌患者中表达上调,5个蛋白在胃癌患者中表达下调.17种差异表达蛋白参与10种生物学过程、9种分子作用和2种细胞组分,涉及的5个生物代谢途径.Western blot结果显示,与正常对照人群相比,代表性差异表达蛋白ITIH4在胃癌患者血清中表达量上调,差异具有统计学意义(1.7517±0.247 vs 1±0,P<0.05).结论:ITIH4可能是诊断胃癌新的潜在生物标志物.展开更多
The spread of tuberculosis(TB),especially multidrug-resistant TB and extensively drug-resistant TB,has strongly motivated the research and development of new anti-TB drugs.New strategies to facilitate drug combination...The spread of tuberculosis(TB),especially multidrug-resistant TB and extensively drug-resistant TB,has strongly motivated the research and development of new anti-TB drugs.New strategies to facilitate drug combinations,including pharmacokinetics-guided dose optimization and toxicology studies of first-and second-line anti-TB drugs have also been introduced and recommended.Liquid chromatography-mass spectrometry(LC-MS)has arguably become the gold standard in the analysis of both endo-and exo-genous compounds.This technique has been applied successfully not only for therapeutic drug monitoring(TDM)but also for pharmacometabolomics analysis.TDM improves the effectiveness of treatment,reduces adverse drug reactions,and the likelihood of drug resistance development in TB patients by determining dosage regimens that produce concentrations within the therapeutic target window.Based on TDM,the dose would be optimized individually to achieve favorable outcomes.Pharmacometabolomics is essential in generating and validating hypotheses regarding the metabolism of anti-TB drugs,aiding in the discovery of potential biomarkers for TB diagnostics,treatment monitoring,and outcome evaluation.This article highlighted the current progresses in TDM of anti-TB drugs based on LC-MS bioassay in the last two decades.Besides,we discussed the advantages and disadvantages of this technique in practical use.The pressing need for non-invasive sampling approaches and stability studies of anti-TB drugs was highlighted.Lastly,we provided perspectives on the prospects of combining LC-MS-based TDM and pharmacometabolomics with other advanced strategies(pharmacometrics,drug and vaccine developments,machine learning/artificial intelligence,among others)to encapsulate in an all-inclusive approach to improve treatment outcomes of TB patients.展开更多
文摘目的筛选腰椎间盘突出症两种不同非手术治疗前后患者血清差异表达蛋白,进一步明确LIDP活血化瘀治疗的血清标志蛋白。方法纳入60例确诊为LIDP并中医辨证为血瘀证的患者,随机分为观察组和对照组,每组30例,观察组给予中医活血化瘀法治疗,对照组用西医非手术综合治疗。基于i TRAQ联合LC-MS/MS分析和鉴定两组患者治疗前后的血清差异表达蛋白质,采用Gene Ontology及KEGG对相关生物信息、差异蛋白的功能和代谢通路进行分析,同时记录治疗前后两组患者VAS及JOA评分,并注意观测是否出现不良反应。结果 i TRAQ联合LC-MS/MS质谱共鉴定到蛋白质组为300个,对应209个非冗余蛋白,观察组符合条件的差异蛋白20个,其中在LIDP血瘀证患者中呈上调表达及下调表达的差异蛋白各占10个,9个差异蛋白序列表达涉及27条KEGG生物代谢途径;对照组符合条件的差异蛋白23个,其中呈上调表达的差异蛋白13个,下调10个,14个差异蛋白序列表达涉及25条KEGG生物代谢途径;筛选出两组共同差异蛋白8个,其中呈上调表达的差异蛋白3个,下调5个,两组共同有6个差异蛋白富集到19条信号通路上,有2个差异蛋白为观察组所特有。生物学信息分析结果显示,代表性差异表达蛋白碳酸酐酶1与转铁蛋白变体在观察组LIDP患者血清中表达量下调,差异具有统计学意义(P<0.05)。两组患者治疗后较治疗前的VAS评分均明显降低,JOA评分明显升高(P<0.05),但两组VAS、JOA评分及总体疗效比较无显著差异(P>0.05)。结论采用活血化瘀治疗血瘀证LIDP可取得与西医非手术综合疗法几乎相同的疗效,治疗是对证的;碳酸酐酶1与转铁蛋白变体是活血化瘀法治疗腰椎间盘突出症血瘀证的目标蛋白质。
基金国家现代农业产业技术体系建设专项资金资助(编号:CARS 24)supported by the Engineering Research Center of EdibleFungi of Fujian Province,National ImprovementCenter of Edible Fungi Varieties(Fujian Subcenter)and the Institute of Biochemistry and Cell Biology,Shanghai Institute for Biological Science,Chinese Academy of Sciences~~
文摘首次利用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)技术结合二维液相色谱串联质谱对草菇(Volvariella volvacea)同核和异核菌丝蛋白质组进行研究。采用QExactive质谱鉴定并经MASCOT软件搜库,对所有鉴定蛋白进行主成分分析(principal component analysis,PCA)、层次聚类(hierarchical clustering)分析和GO(gene ontology)注释,并进行生物信息学分析。结果共获得2335个不同肽段,鉴定到1039个蛋白,其中1030个蛋白具有定量信息。在同核菌丝中显著上调蛋白83个,下调蛋白97个,表明iTRAQ标记技术结合二维液相色谱串联质谱可有效地分离和鉴定草菇菌丝蛋白质组。该研究结果为更深入全面地研究草菇菌丝蛋白质组学奠定基础。
文摘目的:筛选胃癌血清差异表达蛋白,寻找胃癌血清标志物.方法:采集胃癌患者血清和正常对照人群血清各45例,所有血清样本去除高丰度血清蛋白质后,采用相对和绝对定量同位素标志(isobaric tags for relative and absolute q u a n t i t a t i o n,i T R A Q)技术联合二维液相色谱-串联质谱(two-dimensional liquid chromatography/tandem mass spectrometry,2D-LC-MS/MS)分析和鉴定两组间的差异表达蛋白;采用生物信息学对差异蛋白进行分析;用Western blot对蛋白组中的代表性血清差异表达蛋白进行表达验证.结果:iTRAQ标记联合2D-LC-MS/MS分析一共鉴定出10540条独特肽段,对应199个非冗余蛋白,其中符合条件的差异蛋白17个,其中12个蛋白在胃癌患者中表达上调,5个蛋白在胃癌患者中表达下调.17种差异表达蛋白参与10种生物学过程、9种分子作用和2种细胞组分,涉及的5个生物代谢途径.Western blot结果显示,与正常对照人群相比,代表性差异表达蛋白ITIH4在胃癌患者血清中表达量上调,差异具有统计学意义(1.7517±0.247 vs 1±0,P<0.05).结论:ITIH4可能是诊断胃癌新的潜在生物标志物.
基金sponsored by the National Research Foundation of Korea(NRF)Grant funded by the Korean government(MSIT)(Grant No.:2018R1A5A2021242).
文摘The spread of tuberculosis(TB),especially multidrug-resistant TB and extensively drug-resistant TB,has strongly motivated the research and development of new anti-TB drugs.New strategies to facilitate drug combinations,including pharmacokinetics-guided dose optimization and toxicology studies of first-and second-line anti-TB drugs have also been introduced and recommended.Liquid chromatography-mass spectrometry(LC-MS)has arguably become the gold standard in the analysis of both endo-and exo-genous compounds.This technique has been applied successfully not only for therapeutic drug monitoring(TDM)but also for pharmacometabolomics analysis.TDM improves the effectiveness of treatment,reduces adverse drug reactions,and the likelihood of drug resistance development in TB patients by determining dosage regimens that produce concentrations within the therapeutic target window.Based on TDM,the dose would be optimized individually to achieve favorable outcomes.Pharmacometabolomics is essential in generating and validating hypotheses regarding the metabolism of anti-TB drugs,aiding in the discovery of potential biomarkers for TB diagnostics,treatment monitoring,and outcome evaluation.This article highlighted the current progresses in TDM of anti-TB drugs based on LC-MS bioassay in the last two decades.Besides,we discussed the advantages and disadvantages of this technique in practical use.The pressing need for non-invasive sampling approaches and stability studies of anti-TB drugs was highlighted.Lastly,we provided perspectives on the prospects of combining LC-MS-based TDM and pharmacometabolomics with other advanced strategies(pharmacometrics,drug and vaccine developments,machine learning/artificial intelligence,among others)to encapsulate in an all-inclusive approach to improve treatment outcomes of TB patients.