卢大儒：NGs的科学问题与行政管理
卢大儒，遗传学博士，复旦大学特聘教授，生命科学院副院长，遗传工程国家重点实验室副主任，主要从事包括肿瘤分子遗传学；基因检测与基因操作研究。在血友病B基因治疗、东亚人群起源和肿瘤的遗传易感基因与药物遗传学发方面作出突出贡献。先后在Nature、science、Nature Genetics等杂志上发表sCI论文近200篇，承担各类国家和地方科研项目30余项，获得国家和省部级奖励10余项，其中国家自然科学二等奖1项，国家技术发明二等奖1项，省部级科技奖一等奖5项，获得全国优秀教师、全国师德先进个人、中国青年科技奖、国家百千万人才工程国家级人选、全国优秀博士论文、霍英东优秀青年教师一等奖、教育部优秀青年教师奖、上海市优秀学科带头人、上海市高等学校教学名师和国务院政府特殊津贴等荣誉。
邓杏飞：高通量新一代基因测序技术（NGs）在临床医学研究和诊断中的应用
新一代基因测序（NGs）技术正迅猛发展，检测技术日趋成熟，检测费用也不断降低。NGs在医学研究和临床诊断应用方面正在不断扩大。这些都提示NGs正被广泛应用的黄金时光比人们原先预测的来得快。 除了简要分享NGs的技术发展现状，市场应用前景和NGs在临床医学和基因疾病诊断中的重要应用外， 我将特别探讨无创伤产前基因筛查（包括母血游离DNA和完整胎儿细胞单细胞基因检测）和NGs的应用， Ion Torrent基因定型和肿瘤方面的一些最新研究进展和实验资料。
李于：sIRT1 Regulation of Energy Metabolism: Attenuation of Hepatic steatosis and Obesity
Fibroblast growth factor 21 (FGF21) is the hepatocyte-derived hormone that regulates fatty acid metabolism and has potential to treat obesity and diabetes. We recently indicate that hepatic overexpression of sIRT1 in diabetic mice attenuates hepatic steatosis and insulin resistance. However, the in vivo long-term consequence of hepatic sIRT1 ablation in liver physiology remains unknown.
We showed that hepatocyte-specific sIRT1 knockout (sIRT1 LKO) mice with the albumin Cre-loxP system exhibited a striking phenotype with greater propensity for obesity on a chow diet, characterized by increased whole body mass and fat mass, reduced energy expenditure, and unaltered food intake and physical activity. The obese phenotypes of sIRT1 LKO mice were associated with reduced hepatic and circulating levels of fasting FGF21.
Hepatic impairment of FGF21 repressed expression of key enzymes involving fatty acid oxidation such as CPT1α and MCAD, and inhibited expression of ketogenic enzymes including ACAT1, HMGCs2, HMGCL, and BDH1, thereby reducing plasma β–hydroxybutyrate levels in sIRT1 LKO mice. Moreover, transcriptional activity of a FGF21 promoter-driven luciferase reporter was stimulated by sIRT1 activators, resveratrol and sRT1720, in sIRT1+/+ MEFs, but not in sIRT1-/- MEFs.
The ability of resveratrol and sRT1720 to stimulate FGF21 protein was abolished by sIRT1 H335A inactive mutant or by nicotinamide and splitomicin in HepG2 cells. Induction of FGF21 by sIRT1 activators enhanced expression of key enzymes for fatty acid oxidation and ketogenesis.
These in vivo and in vitro findings characterize 1) hepatic sIRT1 as a master regulator of FGF21; 2) sIRT1-dependent activation of FGF21 in liver as a component for adaptive fasting response; and 3) defective hepatic sIRT1 and FGF21 signaling as a key pathological determinant of energy metabolic abnormality and obesity susceptibility.
Erich Gnaiger：Life style and Mitochondrial Competence – Modern Drugs for T2 Diabetes in Aging and Degenerative Diseases.
D. swarovski Research Laboratory (Mitochondrial Physiology), Dept. General, Visceral and Transplant surgery, Innsbruck Medical University; and OROBOROs INsTRUMENTs, Innsbruck, Austria. - Email: erich.gnaiger@oroboros.at
The contribution of mitochondrial dysfunction to the etiology of T2 diabetes and a range of preventable metabolic diseases is the subject of intensive current research with world-wide health implications.
Recently these investigations gained depth and scope by technological advances for diagnosis of mitochondrial function by comprehensive OXPHOs analysis using high-resolution respirometry [1,2]. Fundamental questions of a causal relationship, however, between compromised mitochondrial function and development of T2 diabetes remain to be resolved [3,4] to optimize prevention and treatment of insulin resistance.
For preventable diseases such as T2 diabetes, the evolutionary background of mitochondrial competence provides a solid basis for improved and broad application of a well established modern drug, mtLsD.
Post-industrial societies are characterized by a high-energy input lifestyle with diminished physical activity and high incidence of non-transmittable diseases, in comparison to human populations where physical work is essentially important for sustaining life and in which degenerative diseases (T2 diabetes, various cancers, Alzheimer's) are essentially absent [5]. The capacity of oxidative phosphorylation (OXPHOs) is increased or maintained high by a life style involving endurance exercise and strength training [6].
Life style changes from the age of 20-30 years to the elderly, but is subject to change and intervention. Depending on group selection in cross-sectional studies, OXPHOs capacity declines from the age of 20-30 years [7,8], or is independent of age up to 80 years [9,10].
Independent of age, there is a strong decline of OXPHOs capacity in human vastus lateralis from BMI of 20 to 30 [1]. At a BMI >30, a threshold OXPHOs capacity is reached in human v. lateralis that may be characteristic of a low-grade inflammatory state (‘mitochondrial fever’).
Onset of degenerative diseases (T2 diabetes, neuromuscular degeneration, various cancers) and mitochondrial dysfunction interact in an amplification loop progressing slowly with age, such that cause and effect of mitochondrial dysfunction cannot be distinguished. Diminished antioxidant capacity at low mitochondrial density is an important mechanistic candidate in the state of mitochondrial fever.
For implementing a life style supporting mitochondrial competence and preventing degenerative diseases in modern societies, we need (1) extended research programmes focused on the causative link between mitochondrial competence and effective prevention of degenerative diseases, (2) educational programmes on mitochondrial physiology targeted at general practitioners, teachers and the society at large, (3) cooperation of health care and insurance organizations to support preventive life style activities, and (4) do not miss any opportunity in taking the lead in living the mtLife style Drug (mtLsD).
王煜非：从新版IsO标准修订看血糖监测技术发展趋势
王煜非：上海交通大学附属第一人民医院研究员，中国医院协会临床检验管理专业委员会POCT分委会委员、中华糖尿病学会血糖监测学组委员、上海市糖尿病学会委员、上海市糖尿病学会血糖监测学组副组长、上海市内分泌学会基础学组副组长。
随着人民生活水平的提高和生活方式的改变，糖尿病患病率（尤其是2型糖尿病）在不断攀升。良好的血糖控制有助于预防糖尿病性并发症的发生和发展。准确的血糖检测值可正确指导糖尿病患者的临床用药，饮食和运动，稳定血糖监控，改善生活质量，减少致死致残，降低社会医疗负担。
血糖仪的准确性，精确性和抗干扰能力是准确血糖监测数据的保障。IsO15197（2003版）是国际上针对血糖监测系统评估和验证的标准，近年来，国际组织对此标准进行了大规模的修订并于2013年5月15日正式颁布了新标准，过渡期为三年。
新标准大大提高了血糖仪准确性和精确性的要求，对评估方法亦进行了进一步调整。修订内容主要包括：验证项目定义更清楚，验证标准更严苛，统计方法标准更明确，受试人群分布限制更严格，加大关注清洁消毒过程及临床验证步骤的正确性。不少第三方检测机构研究表明，有一部分血糖仪现有标准也无法全数通过，更难以达到严苛的新标准就。
未来血糖仪的发展趋势是：血糖仪本身技术的突破，具有更好的精准性和更强的抗干扰能力。其他如采血部位的改变，采血与测定一体化，可重复测试的感应技术，用组织间质或体液作为葡萄糖检测的样本，埋入式针头或感应器，无创伤、无样本、红外测定。另一方面血糖数据的管理，联网及远程数据传输等等。
如何提高NGs样品制备实验成功率
新一代测序技术过程中样品制备的质量是测序结果可靠性的根本保障，从核酸的提取、DNA 片段化、末端修复、标记添加，以及产物磁珠纯化、文库构建的每一个步骤，都对最终的实验结果有着巨大的影响。
除了拥有一台高大上的NGs测序仪以及配套的仪器和试剂是远远不够的。很多用户在实验操作中都被繁琐冗长的样品制备过程所困扰，从而严重影响了NGs实验的成功率。其实，从样本核酸的获得到文库构建，每一个步骤都对最终的实验结果产生失之毫厘谬以千里的影响。
在整个繁琐和复杂的NGs样本制备过程中，很多用户都有着诸如此类的疑问： 怎样减少可能出现的手工失误，如何选择合适实验工具进行，怎样避免样本的交叉污染，如何提高珍贵样品的文库构建效果，如何保证RNA样品的测序成功率？
在此次系列NGs网络讲座中，拥有丰富分子生物学产品的Eppendorf 公司将会为各位老师带来涉及NGs样本制备过程中的一系列技巧：
1. 影响文库构建成功率的操作因素；
2. 怎样避免样本制备过程中的交叉污染；
3. 如何保证微量样品文库构建的成功率；
Nanostring--陈巍学基因（19）
本节课程向大家谈一下Nanostring的分析方法。
从以下方面进行阐述：
Nanostring的优点
Nanostring的化学原理
康九红：Pwp1对小鼠Es细胞分化潜能的影响及机制研究
康九红，现任同济大学学术委员会委员、生物学学科专业委员会主任、生命科学与技术学院党总支书记、特聘教授、博士生导师。
Lif/stat3 依赖的信号通路在小鼠胚胎干细胞的干性维持和分化潜能中发挥着重要的作用，然而，它上游的调控机制目前仍不十分明确。我们发现，WD-40 蛋白家族成员Pwp1，对小鼠胚胎干细胞由多能性状态转变成分化状态是必须的。
虽然Pwp1 的下调并不影响小鼠胚胎干细胞的细胞增殖以及凋亡。但是，胚胎干细胞的分化受到了显著的影响，体内和体外实验证实Pwp1 下调强烈抑制了小鼠胚胎干细胞的分化。进一步我们发现，Pwp1 的功能与H4K20me3 密切相关， Pwp1 和H4K20me3 能结合在干性基因stat3 的上游。
下调Pwp1 降低了H4K20me3 在stat3 上游的结合，从而促进了stat3 的表达，进而抑制了小鼠胚胎干细胞的分化。同时，通过ChIP-sEQ 实验，我们也发现Pwp1 和H4K20me3 的确在基因组中具有共同的结合位点。总之，我们发现Pwp1 在小鼠胚胎干细胞的分化潜能维持中具有重要的作用。
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。