Cell and chemical biology of mitosis

Cell and chemical biology of mitosis

纳米生物效应与安全性实验室在2004年首次发现内含Gd原子的金属富勒烯三明治纳米结构颗粒可以直接作为肿瘤的高效低毒化疗药物以来，已经从分子免疫、神经调控、干细胞分化、血管生成等诸多方面对纳米颗粒直接作为高效低毒化疗药物的药效和机制，进行了长达6年多的研究，在国际学术刊物上连续发表了一系列的研究成果，逐渐形成较大的国际影响力。

Simultaneous quantification of 47 gene expression in FFPE samples by a novel PCR-free approach

基因表达(gene expression)是指细胞在生命过程中，把储存在DNA顺序中遗传信息经过转录和翻译，转变成具有生物活性的蛋白质分子。生物体内的各种功能蛋白质和酶都是同相应的结构基因编码的。差别基因表达(differential gene expression)指细胞分化过程中，奢侈基因按一定顺序表达，表达的基因数约占基因总数的5%～10%。

Karen Dell: iBiology:Meet the world's best biologists through the Internet

Karen Dell来自美国细胞生物学学会，她将简述通过iBiology来获取生物学学习和交流的资源。

金颖：Fox3 suppresses NFAT-mediated differentiation to maintain self-renewal of embryonic stem cells

金颖教授为分子发育生物学研究室主任，健康科学中心研究员。金教授介绍了Fox3通过抑制NFAT介导的分化维持了胚胎干细胞的自我更新的机制等前沿发现。

Pluripotency-associated transcription factor Foxd3 is required for maintaining pluripotent cells. However, molecular mechanisms underlying its function are largely unknown.

Here, we report that Foxd3 suppresses differentiation induced by Calcineurin-NFAT signaling to maintain the ESC identity. Mechanistically, Foxd3 interacts with NFAT proteins and recruits co-repressor Tle4, a member of the Tle suppressor family highly expressed in undifferentiated ESCs, to repress NFATc3’s transcriptional activities.

Furthermore, global transcriptome analysis shows that Foxd3 and NFATc3 co-regulate a set of differentiation-associated genes in ESCs. Collectively, our study establishes a molecular and functional link between a pluripotency-associated factor and an important ESC differentiation-inducing pathway.

SureSelect 定制靶向测序--陈巍学基因（23）

Agilent 公司出品的 SureSelect 定制捕获测序 Panel，是一个很好用的目标基因测序试盒。是在肿瘤靶向用药的伴随诊断上的一个先进技术手段。

许多专业的测序公司提供基于 SureSelect Panel 的定制测序服务。

本视频介绍了客户自行定制 SureSelect Panel 的方法。

秦正红：DRAM1 regulates autophagy flux and Bid-mediated cell death via lysosomes

秦正红，博士，教授，神经药理专业博士生导师。1994年在美国宾州医学院研究生院获博士学位，先后在美国国家卫生研究院(NIH)及麻省总医院和哈佛大学医学院从事研究工作。2003年从哈佛大学引进，现为苏州大学医学部基础医学与生物科学学院科研中心实验室主任，中国药理学会生化药理学专业委员会委员，中国药理学会神经药理学专业委员会委员，美国神经科学学会会员。

Damage-regulated autophagy modulator1 (DRAM1), a novel TP53 target gene, is an evolutionarily conserved lysosomal protein and plays an essential role in TP53-dependent autophagy activation and apoptosis (Crighton et al, 2006). However, the mechanisms by which DRAM1 promotes autophagy and apoptosis are not clear. 3-Nitropropionic acid (3-NP) is an inhibitor of mitochondrial respiratory complex II. Intrastriatal administration of 3-NP produces neuropathology resemble to Huntington disease. 3-NP-induced neuronal death was involved in autophagy and apoptosis. In vitro studies with 3-NP in TP53 wt and null cells, 3-NP or CCCP increased the protein levels of DRAM1 in a TP53-dependent or independent manner. DRAM1 induction contributed to 3-NP-induced autophagy activation. Knock-down of DRAM1 with siRNA inhibited the activity of V-ATPase, acidification of lysosomes and activation of lysosomal proteases. Knock-down of DRAM1 reduced the clearance of autophagososmes.

3-NP also induced a transcription independent upregulation of BAX protein levels. Knock-down of DRAM1 suppressed the increase in BAX levels. Co-immunoprecipitation and pull-down studies revealed an interaction of DRAM1 and BAX protein. Stably expression of exogenous DRAM1 increased the half-life of BAX. Upregulation of DRAM1 recruited BAX to lysosomes and induced cathepsin B-dependent cleavage of Bid and cytochrome c release. Knockdown of DRAM1, BAX or inhibition of lysosomal enzymes reduced 3-NP-induced cytochrome c release and cell death.

These data suggest that DRAM1 plays important roles in regulating autophagy flux and apoptosis. DRAM1 promotes autophagy flux through a mechanism involves activation of V-ATPase and enhances the acidification of lysosomes. DRAM1 promotes apoptosis via a mechanism involving recruitment of BAX to lysosomes to trigger cathepsin B-mediated Bid cleavage.

Generating B-lymphoblastoid cell lines using Epstein Barr virus transformation.

Generating immortalized B-lymphoblastoid cell lines via Epstein Barr virus transformation using the B95-8 EBV-infected and producing marmoset cell line.

Western Blot Using The invitrogen NuPAGE Novex Bis-Tris MiniGel System(Aubin Penna.Ph.D)

Western Blot Using The invitrogen NuPAGE Novex Bis-Tris MiniGel System(Aubin Penna.Ph.D)

CellSearch检测CTC--陈巍学基因（30）

欢迎来到【陈巍学基因】,我们这个节目，主要是为大家介绍基因组学，和临床分子诊断的最新技术进展。

今天，会和大家谈一下Jassen公司（强生公司）出品的CellSearch系统。它的主要应用是：检测循环肿瘤细胞，并对癌症给出预后信息。以下是课程内容概括：

1.什么是循环肿瘤细胞(CTC)及其“液体活检”的难点。

2. CellSearch系统的检测原理：（1）用微磁珠对CTC细胞进行富集；（2）用针对DNA的荧光染色剂“DAPI”进行染色，以排除红细胞；（3）区分白细胞和CTC细胞。

3.CellSearch系统实际操作的演示。

4.CTC检测，在癌症诊疗方面所起到的作用。

综上所述：CellSearch系统，是第一个标准化的、半自动化的，循环肿瘤细胞检测系统。它通过快速、精确地确定血液样本中的CTC细胞数量。可以帮助医生在整个治疗过程当中，提供准确的预后评估手段。