祝美国国庆日——自由民主的伟大斗争日, （1944年7月4日《新华日报》社论）

今天是美国国庆日。1776年7月4日，美国人民在华盛顿，杰斐逊等民主主义伟大先驱的领导下，宣布了民族的独立。美国的独立是处于这样的历史条件：它不 但代表美国的民族利益，而且代表美国的民主利益，代表美国要求自由的多数人民而与美国当时的保皇党——大地主、大商人、职业宗教家的集团相对立，这样，领 导独立战争的华盛顿就不但完成了民族任务，同时也完成了政治上经济上的民主任务，而被选举为人类历史上破天荒第一次的民主共和国的第一个大总统了，同样， 杰斐逊总统，不但是独立宣言的起草者，同时也是消灭大地主法案、思想自由法案、全民教育法案、禁止输入奴隶和限制奴隶法案的起草者。这个事实，就说明了美 国独立运动的丰富历史内容，也就说明了美国为什么成为资本主义世界最典型的民主国，而且直到今天，也与社会主义的苏联成为民主世界的双璧。

马克思、恩格斯、列宁、斯大林，这些社会主义的伟大思想家和行动家，对于美国的战斗民主主义及其在世界史上的进步作用，从来都是给予最高度评价的。美国的 战斗民主主义有其光荣的历史传统，美国的独立不是唾手可得，是在八年的对外战争与更长期的对内斗争中完成的。列宁说，这是历史上“最伟大的，真正解放和真 正革命的战争”之一；正因为如此，美国的民主派领袖杰斐逊和杰克逊，美国民主党的这两个创造者，在他们的斗争中甚至被他们的政敌指为“共产主义者”和“赤 化分子”。美国穷木工的儿子林肯，他所领导的黑奴解放战争被马克思称为“开始了劳动阶级兴起的新时期”，而在实际上，他与马克思所领导的美国共产主义者和 欧洲共产主义者也是合作的，他曾委任美国的共产主义者担任他的军官，这样，他就更有理由被当时的反动派指为“共产主义者”和“赤化分子”了，以至最后这些 顽固分子竟然暗杀了他……

民主的美国已经有了它的同伴，孙中山的事业已经有了它的继承者，这就是中国共产党和其他民主的势力。我们共产党人现在所进行的工作乃是华盛顿、杰斐逊、林肯等早已在美国进行过了的工作，它一定会得到而且已经得到民主的美国的同情……

7月4日万岁！

民主的美国万岁！

中国的独立战争和民主运动万岁！

打倒日本帝国主义！

Single Molecule Pulldown can be optimized with Allele’s Fluorescent Proteins

Originally published at AlleleBlog:

In the recent Nature paper, “Probing cellular protein complexes using single-molecule pull-down”, published May 26, 2011, researchers at the University of Illinois at Urbana-Champaign outline their findings for a new method for visualizing protein complexes through a single-molecule immunoassay which combines an antigen capturing chip and TIRF Microscopy. The SiMPull method captures protein complexes and the captured complexes are then visualized with fluorescent dyes or fluorescent protein tags. This is accomplished by using a microscope slide covered with biotinylated polyethylene glycol (PEG) and streptavidin bound to biotinylated antibodies. For single molecule visualization, multicolor labeling provides differentiation of subcomplexes and configurations.

The study includes two validation experiments, where study team members tagged their chosen complexes with YFP, in order to estimate YFP concentration after pulldown and subsequent imaging. They were then able to determine stoichiometric information in human kidney cells, from the isolated monomeric or dimeric YFPs, which exhibited the one and two-step decay responses.

Additionally, in another validation experiment, team members chose to use protein kinase A (PKA) because it’s two catalytic and two regulatory subunits separate in the presence of cAMP. This was accomplished by labeling the catalytic subunits of protein kinase A with YFP and the regulatory subunits with mCherry fluorescent protein. They then used a two-color SiMPull to pull down PKA. After pull-down they imaged the PKA in the presence of cAMP and without cAMP present. The YFP and mCherry signals fluoresced together, demonstrating that the catalytic and regulatory subunits were still attached to eachother. The YFP and mCherry signals did not correlate in the presense of cAMP, reaffirming the fact that the two subunits disassociate in the presence of cAMP. 

Unlike other single-molecule pulldown techniques, the SiMPull does not require purified proteins. It also only requires about 10 cells of sample for protein pull-down and analysis, while traditional Western Blots require about 5000 cells. Moreover, This two-color SiMPull method could be further optimized yielding higher resolution overlay when used in combination with Allele’s mTFP1 and LanYFP, the brightest fluorescent proteins on the market.

The full article can be found at http://www.nature.com/nature/journal/v473/n7348/full/nature10016.html

New Product of the Week: High quality Anti-FLAG Monoclonal Antibody for detection or pulldown, ABP-MAB-DT006, $219/100ug.

Promotion of the Week: save 5% on all of our pre-packaged viruses. Our pre-packaged viruses are all titered at 10^8th or higher, and packaged in 5 tubes for your convenience. To redeem this offer email the code PREPACK to abbashussain@allelebiotech.com.

Making a difference for those in need

From AlleleBlog:

This month 4 major pharmaceutical companies, GlaxoSmithKline, Merck, Johnson & Johnson and Sanofi-Aventis; all agreed to lower the cost of critical vaccines for developing countries.  They have all done this as a part of the international vaccine alliance GAVI.  The companies have severely slashed the prices of vaccines for diseases like rotavirus, a disease that isn’t prevalent in developed countries but causes more than half a million deaths per year.  The model GAVI uses is one where vaccine costs are drastically lowered in developing countries, and this cost is offset by raised vaccine prices in developed countries.
 

GlaxoSmithKline has also reported that they are very close to developing an anti-malaria vaccine, which would be the first of its kind.  This clearly shows a dissent from common pharmaceutical business practice, since Malaria is virtually wiped out in most developed countries.  GSK has no hope of recouping costs for this vaccine by having patients in developed countries pay a premium for vaccination; but this has not deterred their efforts.  Rather they have pledged to make on a 5% profit of the sale of the vaccine which will go toward future anti-malaria drug research.
 

Pharmaceutical companies are often viewed in a negative light for their practice of charging a premium for new drugs.  However, the research, development, trials, and further clinical trials required to bring a drug or vaccine to market are all very costly, somewhat justifying a new drug’s high cost.  Unfortunately this means there is no market for new drugs to combat diseases in developing countries as they cannot afford to compensate drug companies accordingly for their development costs.  This is the key flaw in GAVI’s model, so it is great to see GSK is unhindered by this fact.

Everyday people who work in the biotech field strive to make a difference and help humanity through their research.  Through the work of organizations like GAVI this research can ideally be utilized by all, and not just by those who can afford it.
 

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张洁平: 中国90后先锋改变中国 (节选）

看了下文，感觉北大清华还有未死的精神，甚慰，加惭愧--想起我同代北大人中仍认为“政权是国家机器，维护少数利益集团的利益是应该的”、以及没有正义只有利益、麻烦都是互联网惹出来的（translate: 自焚、钱云会、邓玉娇等等不知道就没事了）的相当普遍的认知水平，不得不感叹时代已经甩开我们前行了。为90后喊一声好！

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中国九零后一代大学毕业正走向社会，其中的精英如清华蒋方舟和北大孙宇晨脱颖而出，自信、独立、忠于自己欲望；网络是他们与生俱 来的工具，在虚拟社会中单打独斗、抱团抗敌，在社交网络中点对点漂移。他们被视为「不是另一代人，而是另一种人」；在武汉向「防火墙之父」扔鞋子的「寒君 依」，在广州反对地铁统一颜色的陈逸华……他们敢于挑战权威和现有秩序，以不同方式塑造中国未来。

一九八九年十月出生的清华女生蒋方舟，言谈之间，不动声色地在身后划了一条线，线的这一边是自己，那一边是比她晚了两个月出生的孩子：「九零后」。

这 个七岁开始写作，九岁出版第一本书，十一岁写长篇小说，在前辈的打量目光中长大的少年作家，偷笑着说，自己终于等来了「九零后成长为一个词」！而她，这时 就可以「忙不迭跳到对岸，站在双手环胸的那一排老朽里，对彼岸的九零后表示出好奇、不解、不满，其中也有一些怵然的敬畏和深切的怜悯……」

可是，用数字来分隔世代看起来有些傻——等等，你没有看出来吗？蒋方舟同学显然是在开一个玩笑啊。这玩笑很符合九零后的水平：有对彼岸「老朽」们的轻微嘲讽，也有与顶着标签的同龄人们的切磋琢磨、惺惺相惜。

怎么看八零后的形象代言人韩寒？蒋方舟说，韩寒很不错，但「没文化」了一点。「他距离真正的精英阶层还很远。」她很直白地讲。

北 京大学大四学生孙宇晨，网络上错写他生于一九八八年，他很介意：「显然不能啊！我是九零年生的，如假包换的九零后啊！」他有接近四点零的本科成绩GPA， 组织过学生社团，竞选过学生会主席，是人人网（类Facebook的大学生中文社交网络）的知名写手，他刚刚揭批了「北大会商制度」，笔下鲜明自由主义气 息的时事评论广受同龄人追捧——无论从哪个方面来看，孙宇晨都称得上足够优秀的北大精英。刚刚拿到美国宾州大学的录取通知书，他已在自己的网络签名档上添 上：美国宾州大学硕士。他从不掩饰自己的优秀。「不选我，他们选谁啊？」聊到竞选学生干部时他会冒出这样的话，带着点调侃，但底气十足。

孙宇晨对韩寒的评价是「精神领袖」。「我小时候还是受韩寒很大影响的。他是我们这一代人的精神领袖，绝对的。」他说，他心目中还有一个精神领袖，是在他出生前一年已成名的北大师兄——王丹。

在说了许多对这两人的印象及他们对他的影响后，孙宇晨补充了一句：「当然，领袖不需要很优秀，领袖只要很像领袖就可以了。」余韵悠长。。。

Generate mouse and human iPS cells with transfected mature miRNAs

http://allelebiotech.com/blogs/2011/06/generate-mouse-and-human-ips-cells-with-transfected-mature-mirnas/

In last week’s blog we discussed generation of induced pluripotent stem cells (iPSCs) with miRNAs expressed from lentivirus. To take it a step further, synthetic, mature miRNAs can be used to avoid the use of viral vectors. Sure enough, Miyoshi et al. published a paper online a few days ago showing that by transfecting 6 miRNAs at 48 hour intervals, they were able to create iPSCs from mouse and human somatic cells. The efficiency is comparable to retrovirus-mediated OSKM factor over-expression (Yoshida et al.), and therefore lower than lentivirus-mediated miR302/369 expression (Anokye-Danso et al.).

In the study of using mature miRNA for obtaining iPSCs, the researchers transfected miRNAs mir200c, mir302s and mir-369 into tissue cultured cells and achieved reprogramming results. Interestingly, only mir302s are common between this study and that with lentivirus-mediated miRNAs by Anokye-Danso et al. There is no current explanation as to why mir-367, which was shown to be required by Anokye-Danso et al., did not seem to be needed in the mature miRNA transfection experiments. Perhaps a level of redundancy among miRNAs, combined with their broad target range and relatively low specificity, allow some of the miRNAs to be interchangeable when used for reprogramming.

Finally, neither of these two recent miRNA-iPSCs works was the first to demonstrate that miRNAs can initiate or facilitate reprogramming. As early as 2008, Lin et al. showed that mir302s could induce pluripotency in a dose-dependent manner by using tet-induced lentivirus expression. They further illustrated that the underlying mechanism is likely through mir302s’ regulation of epigenetic regulators AOFs and other similar factors.

Promotion of the week: Promotion of the week: 10% off on all fluorescent proteins. To redeem, email oligo@allelebiotech.com along with PROMO code: JELLYFISH. See weekly promotions on Facebook.

杨振宁称 结束一党专政不利科研

2011年06月08日

有东方诺贝尔奖之称的「邵逸夫奖」第八届得奖名单昨日公布，七名获奖学者均来自民主的美国和欧洲国家，连续两届未有华人或中国学者得奖。邵逸夫奖评审会主席杨振宁不认同民主是科学发展的元素，又指若中国共产党结束一党专政，将会出现十年整顿期，拖慢中国科学发展。

 

http://www.youtube.com/watch?v=ELk5-mIVDiY&feature=player_embedded 

Wow, now I don't feel so bad about my Beida scientist classmates, after spending most of their adult lives in the US, singing praises to the dictatorship and blaming everything freedom and democracy have to offer. Well, there is still some difference between Yang and my generation's dictatorship-lovers--Yang gets paid now by the dictatorship, whereas the young do not.

又逢六四，写在八九六四二十二周年

又逢六四，自己年龄比当年在广场上的时候已整整翻了一倍了。恰好今天签下购买了靠近陆战一师基地山里面一个四十多 Acre 牧场的合同。暮然回首，六四之后这22年来的日日夜夜，风风雨雨，大大小小，琐琐碎碎，比如可算作在科学上的贡献的论文和专利，还有在自己公司里负担起的十几名员工的livelihood的责任，对社区尽的义务，比如给社区球队的孩子们当足球教练，在“面朝大海，春暖花开”的地方的温暖的家，如今又拥有了70年也不能收回的、不怕推土机强拆的、可以在上面自由地用自己的枪打猎或打靶的260亩山林、溪流地，看着儿子们一天天长大、懂事、又童稚不泯，看着妻子对他们慈爱的目光和偶尔虎妈般的呵斥，听到电话里妈妈的循环往复的旧话题，突然收到老朋友email时的惊喜。。。这一切的一切，都可能在22年前瞬间被一颗子弹变成虚幻，就象发生在那些倒在柏油马路上的学生和市民身上的一样。在一个战场上，倒下的可能是你也可能是我。抛开内心那永远的在枪林弹雨后活下来的人对倒下的人的歉疚感，活下来的有责任不忘记那些倒下的。我们的生命里混合着他们的。记得Saving Private Ryan里Captain Miller 对Ryan的最后遗言“Earn it, earn this…”. 让他自今以后的生活对得起那些在他身边阵亡的战友。

自怜自叹、独自唏嘘之余，忽见一条新闻：

“湖南邵阳男子李旺阳六四时因组织工人运动，六四后被捕判监，他硬骨铮铮拒绝认罪，在狱中度过 21年，上月初刑满出狱时，已被折磨不成人形，抬出监狱时骨瘦如柴、耳聋目盲，要靠亲人喂食维生。他的亲友在网上呼吁外界救助这位「六四硬汉」。朱承志指，李旺阳虽残废了，但脑袋仍清醒，对民主的想念一点不变。”

与他比起来，我的这一生可有显得多么的容易和平淡。
（8964 22周年纪念）