背景:转化医学实现需要临床医师与基础研究者的密切合作。疾病相关的人体生物样本是转化医学研究的重要资源,是实现转化的核心环节。目的:分析临床医师与基础研究者合作开展转化医学研究的必要性,并就临床与基础研究实验室生物样本对接...背景:转化医学实现需要临床医师与基础研究者的密切合作。疾病相关的人体生物样本是转化医学研究的重要资源,是实现转化的核心环节。目的:分析临床医师与基础研究者合作开展转化医学研究的必要性,并就临床与基础研究实验室生物样本对接管理中的主要问题及策略进行了探讨。方法:英文检索词设定为"Translational medical research,biosample,biobank";中文检索词设定为"转化医学,生物样本,生物样本库"。检索中国知网、维普资讯、万方数据、Pub Med数据库2010年1月至2016年12月相关的文献124篇,选取其中部分文献及作者所在单位与其他单位的科研合作经验进行分析探讨。结果与结论:临床与基础研究实验室生物样本对接管理中可能遇到的问题,包括生物样本采集的伦理问题、标准操作流程的设定、生物样本的采集、运输、保存、登记、管理以及有关生物样本的知识产权分割等,医学标准化,高质量的疾病相关的人体生物样本是实现转化医学的核心环节。建立完善的利益共享机制,对于做好临床与基础研究实验室生物样本对接管理,后续转化医学研究工作的顺利进行将起到至关重要的作用。展开更多
微卫星是基因组上的特殊短重复序列,微卫星不稳定的程度与一些肿瘤的分型、治疗和预后相关。目前,微卫星长度变化的检测往往是基于大量细胞,检测灵敏度低。本文整合激光显微切割技术,基于多次退火环状循环扩增(multiple annealing and l...微卫星是基因组上的特殊短重复序列,微卫星不稳定的程度与一些肿瘤的分型、治疗和预后相关。目前,微卫星长度变化的检测往往是基于大量细胞,检测灵敏度低。本文整合激光显微切割技术,基于多次退火环状循环扩增(multiple annealing and looping-based amplification cycles,MALBAC)的单细胞全基因组放大技术和毛细管电泳长度测量方法,经过关键技术点的改进,建立了一套针对组织中少量细胞的多微卫星位点检测方法。研究结果表明,基于技术改进,HE染色的组织细胞经激光显微切割分选后,可成功用MALBAC技术进行全基因组放大,并在多微卫星位点的检测上,获得了高度的准确性和重复性。利用所建立的方法,发现肠型胃癌早期病变组织-肠上皮化生(intestinal metaplasia,IM)的单个腺体内部出现多种长度变化的微卫星改变,说明该癌前病变组织中DNA错配修复系统已经出现问题。本方法所获得的全基因组放大产物,也可以用于外显子组测序和全基因组测序。另外,该方法适用于任何组织的少量细胞,甚至单细胞的多微卫星位点检测,以及全基因组的研究,为精细研究少量病变细胞的基因组特征以及组织异质性提供了有效的方法。展开更多
With the accomplishment of the genome draft sequences, identification of functional elements in genome has become an urgent task. Full-length cDNAs provide an important resource for gene identification and their preci...With the accomplishment of the genome draft sequences, identification of functional elements in genome has become an urgent task. Full-length cDNAs provide an important resource for gene identification and their precise structural feature determination. It also provides a basis for genomic element definition. As many regulatory elements are around transcription start sites(TSSs), precise localization of TSSs in the genome becomes a critical step for identifying the associated core promoters. Massive parallel snapshot of TSSs at a particular time under a specific experimental condition makes it possible to globally analyze important regulatory elements around TSSs and further construct transcriptional regulatory networks. In this paper, we first reviewed two important full-length cDNA cloning techniques: cap-trapper technique and oligo-capping technique. Then,we introduced deepCAGE, a cap-trapper and deep sequencing-based TSS profiling technique, and its applications in the research of transcriptional regulation.展开更多
Fungal infection remains a major problem worldwide, yet treatment options are limited owing to the lack of effective drugs, the significant toxicity of available compounds, and the emergence of drug resistance. The lo...Fungal infection remains a major problem worldwide, yet treatment options are limited owing to the lack of effective drugs, the significant toxicity of available compounds, and the emergence of drug resistance. The low toxicity of calcofluor white(CFW) is an attractive antifungal compound for its known inhibitive effects on trichophyton rubrum and candida albicans growth. However, the efficacy of CFW is limited in most cases. In order to search for effective means to improve its efficacy, using saccharomyces cerevisiae as a model, we have used microarrays to examine the cell's response when treated with CFW on the genome scale. We found that both the PKC-SLT2(i.e, protein kinase C-mitogen activated protein kinase) and the glycogen metabolic pathways are activated upon CFW treatment. These results suggest that the key components in these pathways could be targeted by other drugs to counter the cell's compensative response, thus to further substantiate the inhibitive effect of CFW on fungal growth, which may lead to treatment regimens with improved efficacy of this compound in clinical applications.展开更多
文摘背景:转化医学实现需要临床医师与基础研究者的密切合作。疾病相关的人体生物样本是转化医学研究的重要资源,是实现转化的核心环节。目的:分析临床医师与基础研究者合作开展转化医学研究的必要性,并就临床与基础研究实验室生物样本对接管理中的主要问题及策略进行了探讨。方法:英文检索词设定为"Translational medical research,biosample,biobank";中文检索词设定为"转化医学,生物样本,生物样本库"。检索中国知网、维普资讯、万方数据、Pub Med数据库2010年1月至2016年12月相关的文献124篇,选取其中部分文献及作者所在单位与其他单位的科研合作经验进行分析探讨。结果与结论:临床与基础研究实验室生物样本对接管理中可能遇到的问题,包括生物样本采集的伦理问题、标准操作流程的设定、生物样本的采集、运输、保存、登记、管理以及有关生物样本的知识产权分割等,医学标准化,高质量的疾病相关的人体生物样本是实现转化医学的核心环节。建立完善的利益共享机制,对于做好临床与基础研究实验室生物样本对接管理,后续转化医学研究工作的顺利进行将起到至关重要的作用。
基金the National Natural Science Foundation of China(Nos.1137420,91129000,21273148,91229108,31370750 and 21303104)the National Basic Research Program(973) of China(No.2010CB529205)
文摘With the accomplishment of the genome draft sequences, identification of functional elements in genome has become an urgent task. Full-length cDNAs provide an important resource for gene identification and their precise structural feature determination. It also provides a basis for genomic element definition. As many regulatory elements are around transcription start sites(TSSs), precise localization of TSSs in the genome becomes a critical step for identifying the associated core promoters. Massive parallel snapshot of TSSs at a particular time under a specific experimental condition makes it possible to globally analyze important regulatory elements around TSSs and further construct transcriptional regulatory networks. In this paper, we first reviewed two important full-length cDNA cloning techniques: cap-trapper technique and oligo-capping technique. Then,we introduced deepCAGE, a cap-trapper and deep sequencing-based TSS profiling technique, and its applications in the research of transcriptional regulation.
基金the National Natural Science Foundation of China(No.91229108)the K.C.Wong Education Foundation(Hong Kong,China)
文摘Fungal infection remains a major problem worldwide, yet treatment options are limited owing to the lack of effective drugs, the significant toxicity of available compounds, and the emergence of drug resistance. The low toxicity of calcofluor white(CFW) is an attractive antifungal compound for its known inhibitive effects on trichophyton rubrum and candida albicans growth. However, the efficacy of CFW is limited in most cases. In order to search for effective means to improve its efficacy, using saccharomyces cerevisiae as a model, we have used microarrays to examine the cell's response when treated with CFW on the genome scale. We found that both the PKC-SLT2(i.e, protein kinase C-mitogen activated protein kinase) and the glycogen metabolic pathways are activated upon CFW treatment. These results suggest that the key components in these pathways could be targeted by other drugs to counter the cell's compensative response, thus to further substantiate the inhibitive effect of CFW on fungal growth, which may lead to treatment regimens with improved efficacy of this compound in clinical applications.
