“证候-基因组”的方法学及家系虚寒证的代谢基因表达谱研究
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摘要
背景
人类基因组计划的完成和后基因组时代的到来为中医证候的研究带来了新的机遇,如何实现“证候-基因组学”阐释是目前中医药研究的热点,但“证候-基因组”研究面临着诸多的困惑,故“证候-基因组”的方法学研究对于解决目前研究中面临的困惑具有积极的意义。墨守成规与妄自菲薄都不是科学的态度,本着实事求是的治学精神,为“证候-基因组”研究探索新路。
目的
① 建立“证候-基因组”研究的方法学平台;
② 研究家系虚寒证的差异基因表达谱。
方法
① 理论:系统整理国外基因芯片文献,从理论的角度对基因芯片的实验流程和数据分析进行了系统的分析,为正确应用基因芯片技术提供理论平台。以OMIM数据库为基础,以中医整体观和天人相应观为指导思想,以现代医学模式为参照,建立中西医结合基因模糊分析系统,为“证候-基因表达谱”研究提供临床应用的信息挖掘平台。即从理论的角度对“证候-基因表达谱”进行方法学探讨。
② 实验:从医学的异病同证、遗传学的家系、分子生物学的基因表达、数学上的生物统计、生物信息学的信息挖掘等多学科结合,精选一个家系虚寒证为研究对象,采用直接、间接比较和以药测证3种实验方案,研究虚寒证的差异基因表达谱。以间接比较的差异表达基因为基础建立虚寒证的支持向量机判别模型,同时应用RT-PCR和qPCR的方法对差异表达基因进行临床验证。即从实证的角度对“证候-基因表达谱”进行方法学探讨。
结果
① 系统地分析了实验设计和芯片实验的影响因素,以及21种数据分析方法的优缺点,在此基础上提出选择“证候-基因表达谱”实验方案的建设性意见。
② 建立了中西医结合基因模糊分析系统,该系统是以主题词检索OMIM数据库而建立的本地数据库,其包括现代医学疾病410种,中医证候等词条80条,遗传相关50条,基因性质相关150条,环境相关200条,心理相关150条,社会相关30条,其它130条,共1200条。
BackgroundAfter the Human Genome Project to be completed, the arrival of post genome era provide an opportunity to study the Zheng of Traditional Chinese Medicine (TCM). At present, the important focuses of a research is to explore "Zheng-genomics", in which "Zheng-gene expression profile" confronts a great deal of puzzlements. Therefore, the methodology study on "Zheng-gene expression profile" has positive meanings to clarify therse problemes. It is not scientific attitudes that neither stick to established practices, nor underestimate oneself. We should explore a new way for the study of "Zheng-gene expression profile" in line with the spirit of seeking truth from fact.Objective① To establish methodology platform on "Zheng-genomics".② To researtch gene expression profile on predigree in deficiency-cold Zheng.Methods① Through the reviewing the up-dated references in the world on microarray technology systematically and data analysis, A theoretical fundamental for correct usage of microarray technology may be provided. A fuzzy analysis system on gene functions is set up by combining between traditional Chinese and western medicine from OMIM database. This system adopts the concept of integrity and regards the interaction between man and nature as guidance and refer to modern medicine pattern also. It provides an information platform for clinical application in gene expression related.② According to the conception of different diseases may being same Zheng in medicine, the pedigree in genetics, the gene expression in molecular biology, the biologic statistics on mathematics and information mining of bioinformatics, we adopt 3 kind of experiment schemes to research one pedigree of deficiency-cold Zheng. A justment model of support vector machines is applied successfully for the Zheng by the result of indirect compareing. At the same time, we verificated the difference expression gene by the methods of RT-PCR and qPCR.Results① Through the analyzing the influence agents of experiment systematically, the advantages and shortcomings of the design and 21 kind of mathmatics methods were check out. A constructive suggestion is put forward for choosing an experiment scheme on "Zheng-gene expression profile".② A fuzzy analysis system of combination between traditional Chinese and
人类基因组计划的完成和后基因组时代的到来为中医证候的研究带来了新的机遇,如何实现“证候-基因组学”阐释是目前中医药研究的热点,但“证候-基因组”研究面临着诸多的困惑,故“证候-基因组”的方法学研究对于解决目前研究中面临的困惑具有积极的意义。墨守成规与妄自菲薄都不是科学的态度,本着实事求是的治学精神,为“证候-基因组”研究探索新路。
目的
① 建立“证候-基因组”研究的方法学平台;
② 研究家系虚寒证的差异基因表达谱。
方法
① 理论:系统整理国外基因芯片文献,从理论的角度对基因芯片的实验流程和数据分析进行了系统的分析,为正确应用基因芯片技术提供理论平台。以OMIM数据库为基础,以中医整体观和天人相应观为指导思想,以现代医学模式为参照,建立中西医结合基因模糊分析系统,为“证候-基因表达谱”研究提供临床应用的信息挖掘平台。即从理论的角度对“证候-基因表达谱”进行方法学探讨。
② 实验:从医学的异病同证、遗传学的家系、分子生物学的基因表达、数学上的生物统计、生物信息学的信息挖掘等多学科结合,精选一个家系虚寒证为研究对象,采用直接、间接比较和以药测证3种实验方案,研究虚寒证的差异基因表达谱。以间接比较的差异表达基因为基础建立虚寒证的支持向量机判别模型,同时应用RT-PCR和qPCR的方法对差异表达基因进行临床验证。即从实证的角度对“证候-基因表达谱”进行方法学探讨。
结果
① 系统地分析了实验设计和芯片实验的影响因素,以及21种数据分析方法的优缺点,在此基础上提出选择“证候-基因表达谱”实验方案的建设性意见。
② 建立了中西医结合基因模糊分析系统,该系统是以主题词检索OMIM数据库而建立的本地数据库,其包括现代医学疾病410种,中医证候等词条80条,遗传相关50条,基因性质相关150条,环境相关200条,心理相关150条,社会相关30条,其它130条,共1200条。
BackgroundAfter the Human Genome Project to be completed, the arrival of post genome era provide an opportunity to study the Zheng of Traditional Chinese Medicine (TCM). At present, the important focuses of a research is to explore "Zheng-genomics", in which "Zheng-gene expression profile" confronts a great deal of puzzlements. Therefore, the methodology study on "Zheng-gene expression profile" has positive meanings to clarify therse problemes. It is not scientific attitudes that neither stick to established practices, nor underestimate oneself. We should explore a new way for the study of "Zheng-gene expression profile" in line with the spirit of seeking truth from fact.Objective① To establish methodology platform on "Zheng-genomics".② To researtch gene expression profile on predigree in deficiency-cold Zheng.Methods① Through the reviewing the up-dated references in the world on microarray technology systematically and data analysis, A theoretical fundamental for correct usage of microarray technology may be provided. A fuzzy analysis system on gene functions is set up by combining between traditional Chinese and western medicine from OMIM database. This system adopts the concept of integrity and regards the interaction between man and nature as guidance and refer to modern medicine pattern also. It provides an information platform for clinical application in gene expression related.② According to the conception of different diseases may being same Zheng in medicine, the pedigree in genetics, the gene expression in molecular biology, the biologic statistics on mathematics and information mining of bioinformatics, we adopt 3 kind of experiment schemes to research one pedigree of deficiency-cold Zheng. A justment model of support vector machines is applied successfully for the Zheng by the result of indirect compareing. At the same time, we verificated the difference expression gene by the methods of RT-PCR and qPCR.Results① Through the analyzing the influence agents of experiment systematically, the advantages and shortcomings of the design and 21 kind of mathmatics methods were check out. A constructive suggestion is put forward for choosing an experiment scheme on "Zheng-gene expression profile".② A fuzzy analysis system of combination between traditional Chinese and
引文
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