Wednesday, December 29, 2010

The current hottest post in China internet: 神马都是浮云,唯天地良知永存

----2010年述评 by 1949到2012


20101225,圣诞节。

浙江省乐清市,钱云会死了,死在额定载重量达10多吨的巨型工程车的巨型轮胎之下。这个曾担任过村长的5旬汉子,曾因多年上访被投入监狱的普通农民,终于为他的执着、 坚持付出了惨重的代价,血淋淋的生命的代价。

一场公开的屠杀,在圣诞节的清晨上演。

这一种血腥的罪恶,连耶酥也要双泪长流。这一种疯狂的恐怖,连上帝也要两股战战。

这不是纳粹德国的奥斯维辛,这不是日寇屠刀下的南京,这是2010年的中国。

这是一个血雨腥风的年代,这是一个罪恶横行的岁月。

纵然是万分无力、彻底绝望,我们依然要忍着撕心的痛,吞着哗哗的泪,怒目回视2010。因为,我们要清晰深刻地记住每一桩罪行、每一种邪恶。每一滴 血,每一滴泪,每一滴恨,我们都会收拾起来,存于心底深处。
                                                                       
因为,我们坚信,没有任何罪恶可以永远猖狂。终有一天,我们会将 这一桩桩罪恶,搬上正义的审判台。
                                                                              
一.“李刚时代”
有一个肩杠将星的女歌手,老是在 反复鼓噪:“我们走进了新时代。”唱些什么:“总想对你表白,我的心情是多么豪迈,总想对你倾诉,我对生活是多么热爱。勤劳勇敢的中国人,意 气风发走进新时代。啊,我们意气风发走进那新时代!”

坦率地说,我相信绝大多数中国人,已经很多年不知道“意气风发” 是什么滋味了。我们时常品味的是“垂头丧气、心如死灰、惶恐战栗”。因此,我们真的不敢想象这个“新时代”,会是什么样子的“时代”。

然而当“我爸是李刚”横空出世后,群情共鸣,我们也才恍然大悟,原来这个“新时代”,就是“李刚们的时代”。

如果回顾车轮下的罪恶,人们应该不会忘记“张金柱案”。张金柱,官居郑州一公安分局局长之职。1997年夏,他酒后驾车在闹市区撞死一人,撞伤一人,然后疯狂逃逸, 在很多市民围堵下,才被抓获。其后,新闻媒体包括央视《焦点访谈》等都对此案进行了广泛报道,以致举国关注。最终,法庭以故意伤害罪判了他死刑。

这个案子放到今天,简直是不敢想象。一个公安分局的局长,竟然会因撞死一个人而被判死刑,在今日中国,这不是天方夜谭吗?撞死群众,打伤百姓, 难道不是他们的重要职责、神圣使命吗?邀功请赏都来不及,哪里还敢判刑,竟还敢判死刑?

当然,那个年代也不是什么光辉岁月。可是,其时纲常还没完全崩溃,礼教还未彻底沉沦。那个时候,最最起码,《焦点访谈》还会谈点“焦点”,还有 人关注。
                                                                                 
应该是在2003年的“苏秀文宝马案”上,事情发生了本质的变 化。当年,哈尔滨的权贵富婆苏秀文,仅因街头口角,就怒不可遏地驾起宝马车,朝与之冲突的当事人及围观人群撞去,造成1人死亡,10余人受伤。本来这是一 起比“张金柱案”性质更为恶劣,后果更为严重的案件。其明显涉嫌故意杀人罪和危害公共安全罪,按情节理应求处死刑。然而,即使也是举国围观,群情汹涌,苏 秀文仍仅仅是被依所谓交通肇事罪“判二缓三”,根本不用坐一天牢。

如果回顾这10来年的路,我们可以发现“苏秀文案”是一个标志、一个拐点,是 一个制度彻底沉沦的标志,是一个集团公开宣示罪恶的拐点。苏秀文,这个一向飞扬跋扈的富婆,竟那么“意气风发”地,在哈尔滨的街头,碾着一地鲜血,撞开了 一个新时代。

那一年,是2003年,据说是某个“新政”的开始。

人类社会所建立的任何制度,即使是野蛮血腥的奴隶制度,维系其生命力的关键就是纠错和自我净化功能。好的制度或制度的健康时期,纠错和自我净化 功能较为强大,坏的制度或制度的垂死时期,这方面的功能就很弱,甚至根本没有。

从中国这60年,我们也可看到这种功能的起伏。前三十年,除了杀人就是放 火,谈不上任何制度的纠错和自我净化。后三十年,则可以明显按每十年为一个阶段,分成三段。80年代,国家劫后余生,百废待兴,这个制度还具有较强的纠错和自 我净化功能。民间谣传,在当年的“严打”中,即使是朱德的孙子,也因为轮奸等罪行被杀了头。可见,在功能相对正常的时候,如果威胁到制度的形象和生命力, 这个集团也不会为他们犯下重大罪行的同党兜底。90年代后,则开始了堕落之路。当然到“张金柱时期”,也还是残存有 一定的纠错和自我净化功能。而2000年以后,“苏秀文”则标志着一个制度公然的彻底的堕落,纠错和自 我净化功能已然完全丧失。“李刚门”的制造,则表明此刻正是群魔乱舞的高潮时期。“钱云会案”,更是达到了高潮中的高潮。

必须指出,同样死于车轮底下,“钱云会案”又与前述诸案有很大分别。“张案”和“李案”中,均存在一个从交通肇事向故意伤害或故意杀人、危害公 共安全等行为转化的过程(“苏案”则另有不同,其直接就是故意杀人与危害公共安全,与交通肇事无任何关系),其间,犯罪者虽都有明显的主观故意,但,均应 是处于一种“激情”状态下的犯罪。这些案件中,汽车具有交通工具和杀人工具的双重性质。

而“钱案”,则是有预谋的故意杀人,其间,汽车完全就成了杀人工具。

有人或会说,近些年来被拉下马来的高官也不少,中低级官员更是如 过江之鲫,怎么会说纠错和自我净化功能完全丧失呢?其实,这些官员,不管是哪一层级,也不管是嫖娼、包二奶还是“写日记”等何种原因,他们的落马,都只能 归咎于集团内部的纷争,与所谓法理公平正义无一丁点关系。但,像“苏秀文案”、“李刚门”等案例,则都以鲜血的名义鲜艳地表明,如果站在权贵们对面的是民 众,而非权贵集团成员。那么不管这个涉事官员的官阶高低,即使是李刚这样的副科级;也不管所犯罪行多么深重,即使是公开杀人放火,这个集团都会举全集团之 力进行保护。正如他们一贯以来的德性,根本没有什么原则,也不会讲什么王法、道理,自然也就无所谓什么脸面了。

当然,就像一池湖水一样,如果完全丧失了净化功能,等待他们的命运,也就只有死亡了。

在“我爸是李刚”的时代主旋律中,众多公子千金,那是尽情飞舞,恣意狂欢。宁夏银川的马晶晶马公子、湖南冷水江的曹博文曹公子、福建屏南的陈晨 陈千金,以及海南三亚社保局的诸位公子千金,等等,把所谓衙门,当成了自家的菜园子,自由出入,自由“偷菜”,那是快活得很。

陕西西安的药家鑫药公子,则充分展示了官家公子的不凡风采。年方二旬的他,一双弹钢琴的好手,杀起人来,竟也是那么优雅自如,那么从容不迫。而 西安当局,竟也对这位新科杀手这样呵护有加,明知其杀人后,竟放任他回家逍遥。若不是媒体披露,指不定药公子要继续挥舞着一把染血的匕首,创作出什么宏大 的交响乐来。当然,基于对这个国家本质的认识,对此案是否能得到公正处理,以及“杀人者偿命”这条基本法则是否还被承认,我等并不抱乐观态度。

二.政改迷雾                                                       
温相在就要退休之际,突然就政改数度呛声,果然语惊四座。

虽说最后也表明,这些都只是他老人家一个人说说而已。可是,我们仍然应该对温相表示体谅,表达敬意。在黑暗的夜空,如果闪烁着一丁点亮光,哪怕 只似荧火虫发出的微弱荧火,我们也感到格外珍贵。在冰冷的世界,如果跳动着一丝丝火苗,哪怕只似苇草燃出遇风即息的火苗,我们也感到分外温暖。我们明白, 这些孱弱的亮光与火苗,映透着稀有的良知与坚毅的勇气。

政改如一团团庞大的看不到边际的迷雾,云山雾罩,遮盖中国数十年矣。以至于这个国家,既不知道现在身在何处,也不知道未来要走向何方。在刻意 制造的这一团一团的迷雾中,有的人不但没有困惑,毫不恐慌,反而自得其乐,颇为得意。以为这浓浓的迷雾,就是可以夸耀于万邦的人间仙境。

何清琏女士曾撰文分析,基于特定的历史背景,相比之前历任主事,民众对这两位真的寄予了很大希望,也抱持着很多耐心。然而,在这一幕就要落幕之际,我们不得不表示,人民很失望、很痛苦、很愤怒!

记得80年代末有一本社会影响很大的书籍,何博传先生的《山坳上的中国》,其初版是在1988年。该书将当时危机重重的中国,形象地比拟为身处山坳之上,面 临着何去何从的艰难抉择。当然,一年之后的事变,则给出了明确的答案。他们根本不想攀登高峰,而是无所畏惧地走向深渊。

20余年过去了,如果要非常简约但精准地观察今日中国,那么,我想借 用何博传先生的想像力说一句,现在是“山洞里的中国”。

这个国家分明是携着朝鲜、缅甸、古巴、苏丹和津巴布韦等5个拜把子兄弟,在地底1万米深处(大约等于地狱18层以下),踩着累累白骨,趟着汩汩血水,兴致盎然地去追寻光明 教最终极的真谛----光明神的真身所在。

置身这个阴森的“山洞”,且还要继续锲而不舍地往下钻探。因而,对所谓的“政改”,除了黑暗,我们看不到任何方向。

三.经济疑云
这一年,和近些年来一样,在经济 领域,最大的疑云,就是发改委安的是什么样的居心、在扮演着什么样的角色?

众所周知,发改委历来以抽疯耍宝而著称。但,这一年,该委抽的疯 未免也太离谱了,耍的宝未免也太邪门了。比如,大约是近半年来,发改委是大会小会、人前人后,都在强调要控制物价,控制通胀。巧言令辞之外,甚至还扬言恫 吓,什么要惩治乱涨价的奸商云云。真真是把共克时艰的架势摆得很饱满。

然而,1222日,发改委猛然发出油价上涨令。着实让广大民众在愤骂之余,也 颇为关切和担忧,这个机构该不是超级变态的自虐狂吧?于悠悠苍天之下,于巍巍朝堂之上,自扇大耳刮子的举动真的是那么爽绝了吗?

如果说发改委不是刻意追求变态自虐的快感,那么这只能说明两个问题,其一,国库显然没有他们所鼓吹的那么充实,而且定然相当空虚。以致即使在经济如此焦灼的时刻,他们宁愿顶着不要 脸不要面的骂名,也要施此挖肉补疮、饮鸩止渴之蠢举。按理说,2010年中国的GDP将达37万亿元,经济总量超过日本,成为世界第二。财政收入也是继续保 持二成左右的年度高增长率,将超过8万亿元。国库应该很丰满、很肉感才对呀?不至于会有什么困窘吧?

然而,老百姓们知道这帮爷们是怎么糟蹋银子的。像什么深圳住宅管理中心人均年薪近30万元;什么乌鲁木齐法院采购5万元的按摩椅,黑龙江省公安厅采购4万元的笔记本电脑,苏州交警采购6000元的IPHONE4,抚顺财政局采购2000多元的“U盘”,等等,都只是很小的费用,而且是偶然被披露。更多的那些 银子是如何被挥霍的,如何被搬运到异国他乡去的,老百姓们却只有想像的空间,而没有围观的福气了。

这样子糟贱法,就是金山银山都会垮的嘛。

基于第一个所述国库的问题,那么,我们可进一步推断,发改委的变态自虐,还揭示了第二个问题。即,如果我们不敢说通胀是当局有意制造的话,那么,起码他们也是乐见和鼓励的。

那么多亏空,不通胀,怎么来弥补啊?至于民生的艰难、老百姓的死活,他们才不管哩!

种种迹象都表明,他们是在实施一套通胀的宏伟战略。比如,发改委宏观经济研究院原副院长刘福垣就公开表示,中国要成为强国,必须 有“三高”(物价高、人价高、钱价高)。他竟认为,中国目前没有通货膨胀,物价上涨是拉动内需的一个反应。他还说,“物价不涨也不行,咱们就算涨40年也赶不上美国的物价”。再比如,厉以宁竟然要打破全球公认的3%的通胀率警戒线,认为中国可以承受4.5%

这二位,以及那位说“提高个税起征点于中低收入者意义不大”的财政部 的贾康,就深刻地代表了中国经济界的年度形象。即是:“道貌岸然,胡说八道;祸心昭彰,良知无存。”

尤其是那个厉以宁,出卖了一辈子的良知,都七老八十了,还是乐此不 疲、甘之如饴。只是恐怕他那张斑驳的皮囊下,早已找不到一丝一毫的良知可卖了。

既然如刘福垣所言,要建设发达国家,就必 须要“三高”。那么,显然房价一直居高不下,也应是他们的重要战略部署。难怪,近些年来一直唱空房价的牛刀先生,会显得如此难堪,惹得历来坚定唱多房价的 任志强们是如此快意,狂笑不已。

只是,我们想冷冷地追问:由一个又一个肥皂泡所勾勒出来的景致,难道会持久吗?难道不会瞬间破灭吗?

Thursday, December 9, 2010

Wednesday, November 24, 2010

理想的分量

前不久有幸与大学同学几人聚会,闲聊工作住房、学校钢琴、后院鸡鸭、回国购货之余,偶有提及大陆今日民生之不堪处,非常惊异地发现几人都没有听说过当今流行语“做俯卧撑”、“躲猫猫死”、甚至新红的上过Washington Post的“我爸是李刚“!虽然很理解这些事跟生活在美国的我们没什么关系,实在也没什么理由以为他们会知道这些,但还是觉得有一些惊异:当年以天下为己任、欲救黎民于倒悬、心中充满着理想和自豪的北大学生,会不再关心这些了。或许只是个人的不同,或许只是现在和以前的不同。

提到强拆、上访之苦和不公,有说还不至于混到那个份上吧。但是在公平和正义被肆意破坏的环境里,被不公平、被没尊严不只是谁的“份”或等级的问题了。心中波澜未平之际,看到龙应台回答记者时对读者说得话,顿感些许释怀:“对那已过中年的,我想说,读者和作者在同一个时代里共老了。不坏。对大陆的80后90后,我想说,追求个人的欢乐很好,最壮烈的革命、最伟大的理想,不就是为了让最普通的人得到最寻常的欢乐吗?但是任何一个欢乐派对结束后,总得有几个不醉的人把朋友一个一个送回家。开车的人,决定方向,总得清醒。社会永远需要清醒的人。”一个普通的人追求最寻常的欢乐可能是他能对社会作出最大贡献的最好方式,特别是在正常社会环境里。自认为不是普通人的人,就要担责任,尽努力,敢牺牲。

所幸在我们这一代,经历了89的一代的身后,仍然有有理想者和清醒的人。80、90后留美学生就河北大学撞死女生案给总理温家宝的公开信,在他们收到的签名信表现出有很多甚至中学生已经能看懂社会的不公平,更有青年对理想的理解和孤独的执着;

鲍军良,维也纳大学:
    “我是在奥地利的留学生,现在在德国实习。在网上看到了你们征集签名的有关报道。希望加入。因为我相信,这世界不只眼前的苟且,还有诗与远方。”

这世界不只工作住房、学校钢琴、后院鸡鸭、回国购货(nothing is wrong about those though, but),还有诗与远方!

Monday, November 15, 2010

Integration of Orbigen Products

AlleleBlog: http://allelebiotech.com/blogs/2010/11/integration-of-orbigen-products/

Allele Biotech has been marketing products previously offered by Orbigen since July of 2008. In the process, Allele Biotech has transferred the majority of the Orbigen products to its own product lines, and left out a few that were no longer in demand, under proper license, or in some cases replaced with newer and better versions.
The biggest class of Orbigen products now under the Allele Biotech brand name are the polyclonal antibodies such as anti-EDG1 EDG2… EDG8 (catalogue numbers: PABs 10626, 10619, 10628, 10630, 10496, 1063), anti-BMRPs (catalogue numbers: PABs 10536, 10537, 10538). Other popular polyclonal antibodies from Orbigen include PAB-10983, PAB-10216, PAB-10683, PAB-11651, PAB-11141, PAB-10241, PAB-10469, PAB-10778, PAB-11248, PAB-11665, PAB-10563, PAB-10774. Orbigen antibodies have been tested in hundreds to thousands of labs over more than ten years.
One key product group developed by Orbigen is the baculovirus system, which includes the viral Sapphire genomic DNA and pOrb transfer vector plasmid for efficient packaging. We have further developed the pOrb vector into a bicistronic vector for dual expression or mammalian infections.
Retrovirus packaging cells, on the other hand, have been obtained from non-Orbigen sources and are now called Phoenix 2 as products under a new product line, the Gryphon retrovirus system. These cells have similar functions but were built by different researchers and further selected at Allele Biotech. Like all packaging cells we have experience with, these cells do not attach well after a freeze/thaw cycle. It is absolutely essential to wash away DMSO and better seed them in high quality, attachment enhancing dishes such as our EcoCulture plates.
Products previously under the Orbigen brand name are now included in Allele Biotech’s brand new shopcart (now beta-testing at shop.allelebiotech.com). We hope the new functionalities at the new site combined with the relevant knowledge and information you used to enjoy at our current allelebiotech.com site will provide you with an improved shopping experience.
    New Product of the Week 110810-111410:
Retroviral vector with drug resistance new version, ABP-PVL-IRES10P.
    Promotion of the Week 110810-111410:
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Saturday, November 6, 2010

《南方周末》: “历史深处的来信—四十四年后,终于有红卫兵公开道歉了”

中国 新青年. 于 2010/11/4 22:51:48 发布在 凯迪社区 > 猫眼看人
     今天下班后,我在街头报摊买了最新一期的《南方周末》,头版的标题是“历史深处的来信—四十四年后,终于有红卫兵公开道歉了”。文章报道了一群当年殴打老 师的红卫兵向当年被打的老师道歉。几位被打的女老师现在都80多岁高龄了,当年的学生都60岁了。再不道歉,当年的被迫害者与做恶者就都要太老了。据官方 数据统计,仅在北京市被红卫兵打死的人数就达1772人,民间的统计数字要高得多。这几位老师在暴力狂潮中侥幸逃得性命,而她们的一些同事则进入了死亡名 单。

郭灿辉向《南方周末》记者提及当年对女教师李煌果全部两次伤害的细节:剃阴阳头,从家里揪出来跪在10厘米宽的木板上殴打。 这些事情他从未对家人和朋友提及,“这是地道的耻辱”,在北京魏公村的一家咖啡馆,他终于敞开心扉。

当年参与打人的红卫兵给老师 写信表达了忏悔之情, 84岁的程壁老师接到自己学生的道歉信后非常感动,她在回信中说“你们也是受害者,那时不懂事的孩子跟着起哄,懂事的孩子也有压力,怕跟不上形势,怕犯错 误”。老师原谅了学生,81岁的关秋兰说,账不能算在孩子们头上,道歉固然好,不道歉也会有所反思。半个多世纪过去了,两位老师仍沿用“孩子”称呼已年过 花甲的学生们。

一位学生写道“向那个年代所有死于迫害的人鞠躬致哀,向所有不放弃追求坚强地熬过政治迫害的前辈鞠躬致敬。我们不 想把责任推给别人,只想在自己心灵净化过程中找到一点民族的良知”,另一位女生在信中说“每个人都要认真反思,这一页历史不能就这样不清不楚地翻过去 了”。

很多恶行是难以置信的,已经怀孕的刘美德被红卫兵剪成了阴阳头,强迫她在操场上爬行,把地上的污物强塞到她嘴里,用包有塑 料皮的金属条打她。另一名正在爬行的女教师被一个红卫兵用军用皮鞋碾踩。卞仲耘、胡志涛、刘志平、梅树民和汪玉冰五名教师被活活打死,跪在地上用带钉子的 棒子打死,用锅炉房里提来的开水烫死。美术教员陈葆坤被丢入喷水池中打死,初中女生吴芳芳不小心弄坏了领袖的纸像,被毒打后与陈葆坤的尸体关在一起,以致 终身精神失常。

从申小珂的致歉信中看得出,目前公开忏悔的都是犯错不大,没有命案的人。但更多的人在过去四十四年里保持了沉默。 一些人仍然坚持当年革命的正当性,他们从暴力的行为中得到很大的欢愉。直到现在,不但极少有人向受害者道歉,而且有些人还衷心缅怀那一段时光。

看到这里,我的心被震撼了。我想到那些无辜死难的同胞,想到网上那些死不悔罪的老左派。在乌有之乡和强国论坛深水区,那些沾满同胞鲜血的家 伙对那个罪恶年代无比怀念,在今天竟然把自己装扮成爱国者和所谓的公平正义的代言人,稍微懂点历史的人还能相信他们的谎言吗?

一 位学者问《南方周末》记者“我们对受难的同胞做了什么?我们建立了怎样的记忆?我们是否为他们讨得正义?”。我要说没有,作为一个网友我很清楚。例如,在 强国论坛上,那些老左派不但不悔罪,对坚持正义的网友百般辱骂。他们不承认历史事实,一律说是造谣,百般抵赖。

在真相的基础上走 向民族内部的和解是《南方周末》的希望,这其实也是我一贯的主张。网友们可以看看《南方周末》的这篇专题报道。对我们了解历史,分辨善恶是大有裨益的。

Sunday, October 31, 2010

Cell Cycle Assays-Part I

From alleleblog: http://allelebiotech.com/blogs/

This is the first part of a series of blogs about using fluorescent proteins in cell based assays with established examples, a common theme here at the AlleleBlog.
FUCCI Cell Cycle Sensor
The FUCCI Cell Cycle Sensor is composed of a red (RFP) and a green (GFP) fluorescent protein fused to different regulators of the cell cycle: cdt1 and geminin.
During the cell cycle, these two proteins are ubiquitinated at different time points by specific ubiquitin E3 ligases, which tag them for degradation in the proteasome. The E3 ligases’ activities are regulated temporally and result in the biphasic cycling of GERMINI and CDT1 levels during the cell cycle. In the G1 phase of the cell cycle, GERMINI is degraded; therefore, only CDT1 tagged with RFP is present and appears as red fluorescence within the nuclei. In the S, G2, and M phases, CDT1 is degraded; only GERMINI tagged with GFP is present, resulting in cells with green fluorescent nuclei.
During the G1/S transition, when CDT1 levels are decreasing and GERMINI levels increasing, both proteins are present, so are the tagged fluorescent proteins. When the green and red images are overlaid, nuclei fluoresce yellow. This dynamic color change, from red-to-yellow-to-green, represents the entire cell cycle. This representation can be used to study the effects of elements that may influence cell cycles.
Sakaue-Sawano A, Kurokawa H, Morimura T, Hanyu A, Hama H, Osawa H, Kashiwagi S, Fukami K, Miyata T, Miyoshi H, Imamura T, Ogawa M, Masai H, Miyawaki A.Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. Cell. 2008 Feb 8;132(3):487-98.
CCNB1-CyclinB(NT)-GFP
In late S phage, CCNB1 promoter will be switched on to drive the expression of Cyclin B N-terminus-GFP expression; thereafter the fluorescent signal will be switched off at the destruction box in Cyclin B N-terminus at the end of Mitosis phase. During the intervening phase the fusion reporter protein will translocate from cytoplasm to nucleus by the cytoplasmic retention signal in the Cyclin B N-terminus.
Thomas N. Lighting the circle of life: fluorescent sensors for covert surveillance of the cell cycle. Cell Cycle. 2003 Nov-Dec;2(6):545-9.
GFP-PCNA/YFP-PCNA
GFP-PCNA, a fusion of GFP and PCNA, has been widely used as a convenient tool to monitor the progress of S phase. At the onset of S phase, GFP-PCNA translocates into the nucleus; at mitosis the nuclear envelope breaks down and the nuclear accumulation of PCNA-GFP dissipates.
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Saturday, October 23, 2010

The chairman of the Norwegian Nobel Committee: Why We Gave Liu Xiaobo a Nobel

New York Times, Oct. 22, 2010

我们为什么颁奖给刘晓波

(Thorbjorn Jagland is the chairman of the Norwegian Nobel Committee)

在中国当局对诺 贝尔委员会选择刘晓波(被监禁的政治活动家)作为2010年和平奖得主的谴责无意中说明了为什么值得捍卫人权。

当局声称,任何人都无权干 涉中国的内部事务。但他们错了:国际人权法和标准高于民族国家,而且国际社会有责任确保它们得到尊重。

现代国家制度是国家主权观念演变而 来的,其又是由1648年的威斯特伐利亚和平协议建立的。当时,主权被认为是在一个专制统治者中体现。

但有关主权的想法已经随时间改变 了。美国独立宣言和法国的人权和公民权宣言取代了独裁者控制下的人民的主权作为国家权力以及合法性的来源。

在上个世纪,主权的概念再次改 变了,随着世界从民族主义转移到国际主义。在两个灾难性的世界大战后成立的联合国,让会员国承诺通过和平手段解决争端,并在世界人权宣言中确定全体人民的 基本权利。宣言中说,民族国家将不再有最终的、无限的权力。

今天,普遍人权对世界各地的任意多数提供了一种限制,无论是民主与否。在议会 中的一个多数并不能决定伤害一个少数群体的权利,也不能投票给损害人权的法律。即使中国不是一个宪政民主政体,它是联合国的会员国,而且它已经修改了宪法 以符合世界人权宣言。

但是,刘先生的监禁是清楚地证明,中国的刑法是不符合其宪法的。他被判定犯有“散布谣言,诽谤或者其他手段,颠覆国 家政权,推翻社会主义制度。”但在普遍人权为基础的国际社会,杜绝意见和谣言不是一个政府的工作。各国政府有义务确保自由表达意见的权利——即使说话者主 张不同的社会制度。

这些权利都是诺贝尔委员会捍卫已久的,通过授予那些挣扎着保护它们的人以和平奖,包括安德烈萨哈罗夫为他坚持反对苏联 的人权侵犯,和马丁路德金牧师博士为他争取在美国的公民权利。

毫不奇怪,中国政府已经严厉批评该奖,声称诺贝尔委员会非法干涉其内部事务 和在国际公众的眼睛中羞辱了它。相反,中国应该感到自豪,它已变得强大到足以成为辩论和批评的主体。

有趣的是,中国政府并不是唯一一个批 评诺贝尔委员会的。有些人说,颁奖给刘先生实际上可能恶化中国人权倡导者的境况。

但是,这种说法是不合逻辑的:它导致的结论是我们最好通 过保持沉默来促进人权。如果我们对于中国保持沉默,谁将会是下一个国家要求它保持沉默和不干涉的权利?这种做法将把我们放在一个走向破坏世界人权宣言和人 权的基本原则的道路上。我们绝不能保持沉默。任何国家都没有权无视其国际义务。

中国有充分的理由为它在过去20年来的成就感到自豪。我们 希望看到这一进步继续下去,这就是为什么我们颁发和平奖给刘先生。如果中国是要推进与其他国家的和谐,成为维护国际社会的价值观的一个重要伙伴,它必须首 先给予其所有公民言论的自由。

一个人仅仅因为他表达了他的意见而正在被监禁11年,这是一个悲剧。如果我们要走向阿尔弗雷德诺贝尔所说的 国家的博爱,那么普遍人权必须成为我们的试金石。

Why We Gave Liu Xiaobo a Nobel
By THORBJORN JAGLAND
Published: October 22, 2010

THE Chinese authorities’ condemnation of the Nobel committee’s selection of Liu Xiaobo, the jailed political activist, as the winner of the 2010 Peace Prize inadvertently illustrates why human rights are worth defending.

The authorities assert that no one has the right to interfere in China’s internal affairs. But they are wrong: international human rights law and standards are above the nation-state, and the world community has a duty to ensure they are respected.

The modern state system evolved from the idea of national sovereignty established by the Peace of Westphalia in 1648. At the time, sovereignty was assumed to be embodied in an autocratic ruler.

But ideas about sovereignty have changed over time. The American Declaration of Independence and the French Declaration of the Rights of Man and of the Citizen replaced the control of the autocrat with the sovereignty of the people as the source of national power and legitimacy.

The idea of sovereignty changed again during the last century, as the world moved from nationalism to internationalism. The United Nations, founded in the wake of two disastrous world wars, committed member states to resolve disputes by peaceful means and defined the fundamental rights of all people in the Universal Declaration of Human Rights. The nation-state, the declaration said, would no longer have ultimate, unlimited power.

Today, universal human rights provide a check on arbitrary majorities around the world, whether they are democracies or not. A majority in a parliament cannot decide to harm the rights of a minority, nor vote for laws that undermine human rights. And even though China is not a constitutional democracy, it is a member of the United Nations, and it has amended its Constitution to comply with the Declaration of Human Rights.

However, Mr. Liu’s imprisonment is clear proof that China’s criminal law is not in line with its Constitution. He was convicted of “spreading rumors or slander or any other means to subvert the state power or overthrow the socialist system.” But in a world community based on universal human rights, it is not a government’s task to stamp out opinions and rumors. Governments are obliged to ensure the right to free expression — even if the speaker advocates a different social system.

These are rights that the Nobel committee has long upheld by honoring those who struggle to protect them with the Peace Prize, including Andrei Sakharov for his struggle against human rights abuses in the Soviet Union, and the Rev. Dr. Martin Luther King Jr. for his fight for civil rights in the United States.

Not surprisingly, the Chinese government has harshly criticized the award, claiming that the Nobel committee unlawfully interfered with its internal affairs and humiliated it in the eyes of the international public. On the contrary, China should be proud that it has become powerful enough to be the subject of debate and criticism.

Interestingly, the Chinese government is not the only one to criticize the Nobel committee. Some people have said that giving the prize to Mr. Liu may actually worsen conditions for human-rights advocates in China.

But this argument is illogical: it leads to the conclusion that we best promote human rights by keeping quiet. If we keep quiet about China, who will be the next country to claim its right to silence and non-interference? This approach would put us on a path toward undermining the Universal Declaration and the basic tenets of human rights. We must not and cannot keep quiet. No country has a right to ignore its international obligations.

China has every reason to be proud of what it has achieved in the last 20 years. We want to see that progress continue, and that is why we awarded the Peace Prize to Mr. Liu. If China is to advance in harmony with other countries and become a key partner in upholding the values of the world community, it must first grant freedom of expression to all its citizens.

It is a tragedy that a man is being imprisoned for 11 years merely because he expressed his opinion. If we are to move toward the fraternity of nations of which Alfred Nobel spoke, then universal human rights must be our touchstone.

Thorbjorn Jagland is the chairman of the Norwegian Nobel Committee.

Sunday, October 17, 2010

DNA Repair Pathway Factors in Cell-Based Screening for Restoring Patients’ Sensitivity to Cancer Therapies

Cancers undergoing therapies may develop resistance to treatment. Many current cancer treatments, such as cisplatin, function by creating DNA damage, particularly to fast-dividing cells, i.e., most cancer cells. These treatments may be rendered ineffective by DNA-damage response pathways. Cancer resistance to therapies may come from increased activity in nonhomologous end joining, decreased functions of mismatch repair, or reactivation of the Fanconi anemia (FA)/BRCA DNA-damage response pathway, etc. Ironically the loss of function of some of these DNA-damage repair factors may have partially caused the cancer formation in the first place. Regaining their functions in cancer cells possibly contribute to drug resistance. Molecules that disrupt FA/BRCA pathway or other DNA-damage responses could be used to help restore therapy sensitivity.
Like many proteins that function in DNA-damage repair complexes, FANCD2, a member of the FA pathway factor group, is targeted towards chromatin following damage to DNA in a process called foci formation. There have been recent studies that monitored the foci formation of GFP-FANCD2 in small molecule library screening and identified inhibitors to FANCD2 as candidates for a cancer therapy sensitizer. The assays can be improved in a number of ways. There are fluorescent proteins (FPs) that are much brighter than EGFP for increased sensitivity. For instance, the monomeric green FP mWasabi is about 2-3 fold brighter than EGFP, with narrower emission peak, and is more stable under acidic environment. The newly developed lancelet YFP (LanYFP, developed/introduced by Allele Biotech) is astonishingly 10 times brighter than EGFP. Since it has a longer excitation and emission wavelength, it should inherently have a better signal to noise/background ratio compared to EGFP because cells autofluoresce less in long wavelengths. The improved brightness would also help in this respect. The fold difference between foci and LanYFP background will be the same as EGFP, but the contrast will still probably be better because of less autofluorescent background and significantly higher fluorescence reading in foci.
Other factors that may be used as a screening target when fused to effective FPs may probably include:
1) Homologous recombination (HR)
a. End Resection
MRN complex (MRE11, RAD50, NBS1)
CtIP, RPA, ATM, ATR, Exo1, BLM, RMI1, TopIIIa, DNA2, BRCA1
b. Synapsis
RAD51, BRCA2, PALB2, RAD51B, RAD51C, RAD51D, RAD51AP1, XRCC2, XRCC3, RAD54, RAD54B
c. DNA synthesis
DNA polymerase delta, PCNA
2) Nonhomologous End Joining (NHEJ)
Ku70/Ku80, DNA-PK, Ligase IV, XRCC4, XLF
3) Fanconi Anemia Pathway
FANCA, FANCB, FANCC, FANCE, FANCF, FANCG, FANCL, FAAP100, FANCM, MHF, FAAP24, FANCD2, FANCI, FAN1, FANCN, FANCJ, FANCM
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Original post: http://allelebiotech.com/blogs/2010/10/dna-repair-pathway-factors-in-cell-based-screening-for-restoring-patients%E2%80%99-sensitivity-to-cancer-therapies/

Friday, October 8, 2010

Dr. Liu Xiaobo Wins 2010 Nobel Peace Prize

For "his long and non-violent struggle for fundamental human rights in China".

"From my personal angle, I feel in a dictatory society if you want to be a person with dignity, if you want to be a honest person, fight for human-rights improvement, fight for free speech, being ... [in prison] is part of what you are undertaking, and there is nothing to complain," Liu Xiaobo told CNN in 2007, while he was between a series of house arrests."

"Since you chose to do this, you must have a preparation for being in prison," he said. "Entering the prison you must face these things peacefully, not complain [about] others. I even don't complain [about those] ... who arrested me, because this is their inevitable action. I can also not let them arrest me if I chose other way." said Liu before the current prison sentence.  He served one previously for the 89 student movement.

He lost his freedom, willingly, twice, for the freedom of many others.  He is a living example of honor, dignity, bravery, and idealism.  Proud to have shared battle field with the hero 21 years ago.

Wednesday, September 29, 2010

Congress may let SBIR authority lapse this week

From AlleleBlog: http://allelebiotech.com/blogs/2010/09/congress-may-let-sbir-authority-lapse-this-week/

The SBIR/STTR/CPP now appears likely to expire on Thursday night, September 30.
Some will deny it but here’s what’s happening.

Allegedly the Senate and House were close to a compromise complete with an 8 year
reauthorization of SBIR/STTR/CPP but each time it goes back to the House (Nydia &
Day), they change the VC language to masquerade 100% VC involvement as a compromise.

Because time is so short, the Senate passed a bill (S.3839) to simply extend
SBIR/STTR/CPP through January 31, 2010. The House was going to pass it on Wednesday
with the President signing Thursday. However, the word on the street is that Nydia
Velazquez, chair of the House Small Business Committee, and her illustrious second,
Michael Day, are rejecting the bill and are poised to let SBIR expire if necessary,
at least in the short term.

It seems that Velazquez’s hope is to move the SBIR reauthorization into the lame
duck session and incorporate all her Wall Street investors’ 100% non-compromise VC
ownership and jumbo award support into a must pass, end of the year omnibus bill
that can’t be touched by her detractors.

This sounds like a script for TV, but several years ago we had a similar year end
omnibus situation involving Nydia (as ranking member) and Sam Graves (subcommittee
chair) and BIO/NVCA, but the main difference was that the small business committee
chair was Donald Manzullo who nipped it in the bud. In our scenario today we have
to look to the House leadership to do it, but it will take your involvement.

Many senior people in the democratic party called for the House to support the
Senate compromise bill H.R. 2965, but Nydia ignored those calls, as did Jason
Altmire, the creator of this infamous Altmire Quagmire. Now Nydia’s really “miffed”
because last week she tried to “scrub” H.R.5297, the Small Business Jobs Act of
2010, but the Obama administration and Speaker Pelosi rolled her over and passed it.

CALL TO ACTION

If SBIR is important to you and your company, it’s time to get serious and realize
that this program can, and will go away unless you make a big noise to let your
politician’s know how you feel. All of us are sick of this, and we’re now facing a
lapse. Eight times this program has been deemed important enough to keep going (via
a CR) but will Nydia be successful in blocking this ninth attempt?

Voting will occur in the House on Wednesday and this may be the last time until
after the election that the SBIR extension bill could voted on. That means we must
act on Tuesday, September 28.

Here are some suggestions and rationale behind them.

CALL CALL CALL the House Tuesday September 21! Call Nancy Pelosi’s office at (202)
225-4965, Steny Hoyer (majority leader) at (202) 225-4131, Nydia Velazquez (202)
225-2361, also the House Small Business Committee line (202) 225-4038

Those of you who are good democrats, call the remaining House Democratic caucus
leaders: John Larson 202- 225-2265, Xavier Becerra 202-225-6235, Jim Clyburn
(202)225-3315

Those of you who are good republicans, call John Boehner (202) 225-6205, Eric Cantor
202-225-2815

Tell them in your own words that SBIR is about to expire and is being held hostage
by Nydia Velazquez. Let them know how important continuation of SBIR is to your
business and the country. Ask them to please support S.3839 (additional temporary
extension of programs under the Small Business Act and the Small Business Investment
Act of 1958) to keep the program from lapsing this week.

I realize that I’m asking you to do something that requires a good chunk of your
time. However, at the risk of losing you as a reader I must tell you that I donate
a large share of my time to try and keep you informed about this program, and I’m
not asking you to do anything for me, only for you and others like you. We do have
some good representatives from both parties BUT they need to hear from you and
quickly.

If you’re bold ask, “I would like to know how a party can let itself get hijacked by
a few people (like Nydia) on a vitally important, highly regarded and accepted
program. This action is to the detriment of your constituents, the country, and
yes, even your own party!”

Here’s what’s going on in the back rooms (formerly smoke filled) The Senate agreed
on a 4 month extension for SBIR because they (Senate) largely (including many on the
Republican side) did not feel a reasonable bill could be passed in the lame duck
session. The Senate has offered up some huge compromises that some believe even
James Greenwood from BIO could live with. The very long shot is that with enough
pressure we might get a compromise bill passed by Thursday.

WHAT HAPPENS IF SBIR LAPSES, EVEN FOR A SHORT TIME

This is an interesting question. Theoretically those projects (grants and
contracts) that are already in place should be okay, but some not. All new unsigned
agreements would stop. Agency comptrollers may start adjusting their budgets to put
the overall 2.8% SBIR/STTR back into their own research pools. Administrative
funding for SBIR could be severely cut back. Remember, all of your grants and
contracts are “subject to the availability of funding.”

On the other hand, SBIR can be voluntary, so some agencies may choose to keep their
SBIR doors open, hoping for, or expecting the reinstatement of the program.

In any event, this is bad for you and the agencies.

The Insider will be on the Hill Wednesday and Thursday, so we’ll do a follow up
report to you asap.

Rick Shindell
SBIR Gateway
Zyn Systems
40 Alderwood Dr.
Sequim, WA 98382
360-681-4123
rick@zyn.com
www.zyn.com/sbir

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Sunday, September 26, 2010

Dealing with Interferon Response When Doing RNAi

From AlleleBlog: http://allelebiotech.com/blogs/2010/09/dealing-with-interferon-response-when-doing-rnai/

Off-target effects are a major problem when using RNA interference (RNAi) to silence genes in mammalian systems. One potential source of off-target effects, by either transfected siRNA duplexes or transcriptionally expressed shRNAs, is the inadvertent activation of the interferon response. There are several steps that can be taken to deal with this problem.

Delivery
Interferon response is more likely when high levels of siRNA are used; it is important to transfect the minimum amount of the siRNA duplex that gives rise to a specific RNAi response, as assessed by the level of expression of the target mRNA and/or protein. The level of stable shRNA expression achieved by using lentiviral or retroviral vectors is comparatively modest. Unless very high levels of shRNA expression are achieved, for example, by using highly transfectable cells and a very efficient shRNA expression plasmid, nonspecific activation of the innate immune response are less likely to be induced.

Design
Previous work has shown that the interferon response is induced by dsRNAs of ?30 bp in length and that perfect dsRNAs of as little as 11 bp in length can produce a weak induction. One possible approach to solving the problem of nonspecific activation of the cellular interferon response is to design the siRNA duplex or shRNA precursor so that it does not contain any stretches of perfect dsRNA of ?11 bp.

Detection
If activation of the interferon response remains a concern, it is possible to routinely check for this effect during the course of an RNAi experiment. Analyzing the level of expression of an interferon-response gene, such as oligoadenylate synthase-1 (OAS1), interferon-stimulated gene-54 (ISG54), and guanylate-binding protein (GBP), in the transfected or transduced cells by northern blot or RT- PCR assays are commonly used.

Can there be any more convenient alternative method for checking interferon response? One potentially useful product could be HiTiter™ pre-packaged lentiviruses that would have a fluorescent protein (mTFP1, mWasabi, or the brightest FP in lanYFP) under the control of an ISRE (IFN-stimulated response element) or GAS (IFN gamma-activating sequence)*. This could be another group of Product-on-Demand type of reagents, meaning that we will have the design ready, but only to produce them upon ordering. This way the cost to us and the price to customers can be kept at minimum.

To read the wholr blog, click here.

Sunday, September 19, 2010

韩寒: 游行的意义

在jiu月18日这个敏感的时刻,我有的朋友开始研究要不要you行。当然,游的主体可以是反ri保diao救船长。终于,在一个很多论坛里连“you 行”两个字都打不出来的国家里,我们有行可以游了。那么,要不要参加这次命题一日游呢?

首先,我认为在现代中国社会中,分为三个阶级,那就是主子,奴才和狗,而我们往往一人饰两角,至于饰演哪两个角色,我想不会有人觉得他在演主子吧。前一阵子,主子需要奴才去附和和伺候,但是现如今,主子需要狗去吼两声,因为在狗的逻辑里,无论主子怎么对待它,只要有外人来犯,狗总是该看家护院的。

当弄明白了这个以后,回头想想就容易多了。但是,在这三个阶级以内,好在我还有选择做花花草草的权力。我的选择依据是,对于相关部门,小事和大事他们的区别就是抗议一次和抗议十一次,有特权有能力的地方尚未出力,除了把人家日本大使变成了应召男郎以外,我们相关部门情绪稳定,并不见什么实际决心,别说武力上,连经济上都不敢有所动作。他们韬光养晦,所以我也韬光养晦。毕竟,我等做狗也罢,但要做一条戏狗,情以何堪。

纵观事态发展,领dao的内心似乎并不愤怒,领dao只是觉得窝囊,那自然,我们也只能跟着觉得窝囊,你哪有上街去表达窝囊的,那岂不是更窝囊。领dao没面子的时候,我们给他们长脸,但领dao有面子的时候,我们被他们掌嘴。我被欺负,我不能游,你被欺负,你让我游,我又情以何堪。你也别说这种民族国土大事应该是我们一起被欺负了,就算政fu不作为,你活的一塌糊涂,也应该挺身而出。我自然可以挺身而出,但我的第一主题就是要求政fu去作为,第二主题才是控诉来犯者,因为领土问题从来都不是老百姓能解决的和该去解决的,尤其是在我国,老百姓自己都没有一寸土地,,所有的一切,都是问政fu租的,所以,理论上,这事对我来说,就是我的房东在和别人就一块在地上的瓦而争执,这块瓦的确是风大的时候从房东的房顶上掉下来的,但房东也不敢去捡,因为可能要和隔壁人家打架。那我等租客在里面搅和什么呢。无土地者要去为他人争取土地,无尊严者要去为他人捍卫尊严,这样的人多少钱一斤?一斤多少个?

但毕竟,这样的you行安全,好玩,显得很酷,关键是游完以后还能正常工作学习,甚至还有助于未来发展,毕竟也算不容易,所以大学生和老百姓抱着尝鲜唱黑脸的角度去游一游无妨。到时候政fu唱一个白脸,说不定能有所见效。况且现在去you行玩的人相比起以前you行玩的人也有着些许不同,以前是彻底的国政不分,被卖数钱,现如今很多青年终于能够将所谓爱国这件事情想的更明白,他们虽然依然愤怒,但开始反思自己为何每次都是那么窝囊和被动,回头也能更客观的看待国家和政fu的关系,这也算是一个进步。对于任何国家来说,国家就是一个女人,zhi zheng者就是占有她的男人,有幸福美满的,有相处和睦的,有家庭暴力的,有关系紧张的,有离婚再嫁的,有不能改嫁的,但无论如何,你爱一个女人总不能连她的男人也一起爱了去。

最后,这些都不重要,最重要的是,我,如果今天能为唐fu珍,谢chao平而you行,那么明天我就一定会为钓 yu岛和奥运火炬而you行。但这又是一个悖论,往往你能够为唐fu珍,谢chao平you行的时候,你往往就不会有钓yu岛奥运火炬之类的事,而且更不会有唐fu珍谢chao平之类的事出现。一个对内不能和平you行的民族,他的对外任何you行是完全没有价值的,那只是一场集体舞。

Sunday, September 12, 2010

韩寒:保住非法字符

有朋友问我,钓鱼岛事件你怎么不发表一点意见,谴责一下日本。我说,虽然脚下一片自己的土地也没有,但对于领土的问题,我也是很在意的。最早的时候,我在一个论坛上看见这事,我义正言辞的写了一句,“保住钓鱼岛”,结果该论坛告诉我,我试图发表非法的内容,请修改。我百思不得其解,直到把帖子改成了“保住尖阁列岛”,这才顺利合法的发表了。

这的确是一件大事,外交部都周末加班破例追加谴责。如果大家一切都好,生活如意,老婆孩子房子车子工作休息健康医疗一切都能保住,闲情雅致之下,民族情操之下,又不愿韬光养晦,当然可以追保钓鱼岛。但是如果你自己还有什么保不住的,先把自己的保住再说,不要操那么前卫的心。也许你会说,在大是大非之下,你个人的小失小患算的了什么,是的,不过每个人都有其自定义大是大非的权力。比如这种事情,我认为先要看政府的态度,你怎么能冲在领导的前面呢?领导表示谴责,意思就是让你表示谴责,领导表示遗憾,意思就是你可以谴责完毕了。领导要谴责,你要动手,这是领导能容忍的极限,你如果真的动手,领导就要惩罚你了,因为领导在下一盘很大的棋子,你身为一个棋子,怎么能跳出棋盘了呢?而在这盘棋子里,你是一颗黑棋,领导就是一颗白棋,一来因为劳动人民总是黑点,而且也容易变成黑户,黑,是你最贴切的颜色,但最关键的是,已经洗白领导要求你在冲锋的时候出来唱黑脸,而领导在关键的时刻唱白脸。事后弄不好你还能发现领导和来犯者还在欢快的谈一笔大生意。

钓鱼岛的问题,我相信我们官方更看重的是自己对内是否稳固,底下石油倒不是那么有所谓,那些都是日本人要的,这也是他们在70年代重新对钓鱼岛起邪念的原因,而中国政府只要稳定,不要在外交和军事上有任何未知的风险,所以这导致了这个本来不复杂的问题一定将被拖延成一个非常复杂的问题。
在我国的版图里,类似有争议有可能引起国家间任何摩擦和局部战乱的地方,只要别太大块,改变了中国是一只鸡的形象,而公众和舆论也不是太了解的犄角旮旯,官方可能觉得让一点也就让一点了,就当卖给地产商了,公鸡母鸡实在是没那么所谓的。钓鱼岛因为一直一来比较有名,公众关注度高,尤其是看了多年新闻联播,领导人都是在钓鱼台国宾馆接见外宾,搞半天钓鱼台都归了别人,这太没面子了,所以,钓鱼岛是政府事关领土问题的形象工程,是底限,我相信这个应该不会让给人家。而对中国政府来说,最好的解决方法就是一直拖着,拖到地壳板块再次发生移动,钓鱼岛直接镶到了福建省,这下就省事了,什么海域石油的事再说。所以我也不担心钓鱼岛被日本人占去,虽然事实上他们的确快占去了。而这次的事件,最好的结果就是船长要被关押十天,在我们强烈谴责严重抗议了九天以后,日本把人放了,我们也算终于抗议有了成果。至于唱黑脸的人们,当然闲来无事唱唱也无妨,只是不要入戏太深,不要影响到自己的生活,不要忘了家人和自己所应该拥有的一切更应该保,不要为发现你已经干了而领导连酒瓶子都没开而伤心,也不要以为自己真的在急民族所最急,这个民族,总有更急的。
http://blog.sina.com.cn/s/blog_4701280b0100lcum.html

3 Ways of Making DNA Libraries through Oligo Synthesis

http://allelebiotech.com/blogs/2010/09/3-ways-of-making-dna-libraries-through-oligo-synthesis/

Pools of DNA molecules of related but non-identical sequences are often used for selecting cDNAs that encode polypeptides with desired functions (such as in antibody screening), or DNA segments as protein binding sites (through SELEX), or DNA molecules that can catalyze reactions (DNA enzymes or deoxyribozymes), etc. The most direct way of creating DNA libraries is to introduce mixed bases during the synthesis of the oligos that will be used in creating the libraries.

1) The most commonly used method of generating degenerate oligos is to use mixed phosphoramidites (aka amidites, the building blocks of oligo synthesis) at desired positions in an oligo, e.g. using “N” to incorporate dA, dC, dG, and dT nucleotides, or “Y” for pyrimidines, “R” for purines. Mixed base oligos from most oligo suppliers are simple to order (and at no extra charge from Allele and a few other sources). During automated chemical synthesis of oligos, the synthesizer consecutively adds dT, dA, dC, or dG in the case of “N” at a pre-set ratio (e.g.25% each). This procedure does not always result in expected usage of each amidite because different amidites have different coupling efficiency, and the order of addition may also bias against amidites that are added later.

2) Using mixed bases like in method 1) leaves little control to achieve ratios of codons for specific amino acids. On the other hand, by using trimer amidites, which can be used for adding 3 nucleotides in each synthesis cycle, one can create oligos encoding selected amino acids at pre-determined percentages. However, this procedure is difficult to perform because trimer amidites are bulky and hard to couple to the elongating oligo; any moisture present during synthesis would have even more severe adverse effects than with regular amidites. Trimer oligo synthesis projects cost several thousand dollars per oligo on materials alone, and the risk is quite high that the oligos would not turn out of desired properties and qualities. For commercial users, this process has another problem—it is patented.

3) Another method for making library oligos is the so called “split-and-pool”, which is particularly suitable for having diversified amino acids embedded in otherwise common sequences like the CDRs within antibody variable regions. The latest oligo we made last month was a ~72 nt oligo with 8 locations that have pre-determined composition of amino acids, i.e. 20% Ala, 10% Gly, 12% His, etc. The procedure took us about 8 hours and we estimated the cost to be about $1,000. The subsequent sequencing results confirmed that ~70% of the clones using this oligo have desired degeneracy, compared to a similar oligo made by a bigger oligo company, at only 40%. In addition, we did not see any stop codon interruptions or major abnormalities.

DNA pools can also be generated by error-prone PCR, or more specifically with overlapping PCR using degenerate primers. The bottleneck for a library screening is how to handle big enough a number of colonies to accommodate the population, e.g. 10e10, or at least 10e8 clones are needed for finding high affinity antibodies. The second critical point is to have a robust and consistent selection readout such as fluorescence in cell sorting.

New Product of the Week 090710-091310; loxP-mWasabi reporter T cells, email vivec@allelebiotech.com for details.

Promotion of the Week 090710-091310: 15% off our NEW purified fluorescent proteins (not plasmids); All Expressed from E.coli PROMOCODE: 090910FP

Thursday, September 2, 2010

[转贴]韩寒:草莓

It's been a while since HH published new post, still sharp, still deep, still alive.

http://blog.sina.com.cn/s/blog_4701280b0100l4sf.html

这几天应该是独唱团第二辑上市的时间,由于最近台风登陆,所以还是有所延误,我们在为更好的质量和空间而努力,也为使他变成合法的月刊,而不是居无定所的绝唱团。在这个期间里,GQ杂志为我颁发了一个传媒人奖,这两年,无论是以往的《时尚先生》,还是创刊一年的GQ,都让时尚杂志的男刊不再难堪,很多其他媒体禁忌的名字,甚至出现在他们的名册里,GQ还曾被半路召回,这些都不是业内笑话,而是各种尝试,其实这些都应该是一个传媒人应该去做的。什么是做一个传媒人,我深深的思考过,在我们国家,其实就是做一个传达一下领导意思的媒人,做的不好就送你一个传票,然后就挖煤,这就是传媒人。在很长的一段时间里,我其实是一个被传媒的人,今年,我成了一个所谓的传媒人。好在这个奖项是年度传媒人,它并没有说年度最成功传媒人,我自认为是年度最失败传媒人。失败的原因,以后我会和你们讲来。我们知道,其实很多人宣称的“办一本杂志”“做一份报纸”,都是理想化的称呼,从程序上,这些都是非法的,合法的说法是——某个拥有党委的国有出版社或者杂志社聘请你和你的团队来打理一下他们的杂志。当然,唯一值得欣慰的是,没有我们,他们通常自己的打理的不大好。传媒是一个很大的名词,无论是传统的图书,出版,杂志,报纸,电视,电影,广播,互联网,电子阅读,甚至我们的耳语,打一个电话,贴一张海报,都是传媒,传媒的影响如此之大,谁都想控制它。但传媒其实不该控制在任何人手里,他应该是一片开放的天地,只要善待,谁都可以使用和拥有它,它的上司只有一个,法院,它的罪名只有一种,诽谤。这便是我理想中的传媒。可是理想是每一个人都会说,大家都爱听的,就好比你我都愿面向大海,春暖花开,说一次心里爽一次,却始终无法走出脚下的泥泽。但有念想的人总能走得更远,求生欲强的人总能活的更长,所以,我始终是高兴和乐观的。不过,作为所谓的传媒人,我们总需要不断尝试,我本该很高兴的去北京讲这些话,但因为实在不能过来,所以由老朋友黑狗达代我领一下奖,另外向现场的主编王锋先生,左小诅咒先生,姜文先生,刘北宪先生,白岩松先生和柴静女士问好。向程益中先生问好。

也向刚刚被跨省刑拘的谢朝平先生问好。

这些天一直没有更新这里,我发现在中国写杂文其实是一件很痛苦的事情,虽然这个国家给我们提供了大量的杂文素材,但是你突然发现,写了一阵子以后,当再有新闻出现,你恨不得直接写一句,观点请参考我某年某月写的某一篇。我们的领导群从这一批换成了那一批,治国口号从这一堆换成了那一堆,丰功伟绩从这个会变成了那个会,社会悲剧只是从这个人变成了那个人,换人不换事会让写作者觉得很痛苦,因为大部分的作家都讨厌反复阐述,结果事儿又是反复发生,对我们的遣词造句提出了很大的要求。因为我真的想一直继续写下去,不想让自己厌烦和麻痹。

昨天去看了《盗梦空间》,向大家推荐。

Friday, August 27, 2010

[转贴]李承鹏:成龙为什么不憎恨

成龙很久没拍过好的电影了,无需拍,因为现在他本身就是一部好的电影。从“中国人确实需要管一管了”到向菲律宾伸手“友谊之手”,他完成人生演技最重大的一次提升,演得太好了,下一步只有出演特首才承受得了。



我并不同意因为自己人被欺负,就要派解放军去轰炸菲国把菲佣弄到大巴上一一击毙最好把东南亚全部解放了,一是因为那儿不归我们解放,我们只有更加文明才不会被野蛮欺负,用野蛮对付野蛮只有更加野蛮(比如非洲有些地方现在还杀来杀去),再就是,如果中国人被欺负就要轰炸,就得轰炸强拆过中国商人店铺的某某国还要轰炸让中国民工淹死在海滩上的某某国更要轰炸把两个中国商人活活打死的某某国,这要花费太多的炸弹……当然,这样的报道经常是被屏幕了的,因为我们必须是大国,小民,而大国。



但成龙说“如果警方一开始就击毙绑匪,人们会说怎么不谈判?如果警方先谈判,人们又会说怎么不一枪击毙”,我估计好多人读到这里咯登一下,因为确实,前段时间幼儿园事件我们曾批评过警方不谈判就崩了没什么实质威胁的劫匪,不人性,这次菲国的例子正好反过来,让你有些无从反驳。从这一点看来,说成龙是霸王洗发水用多了,不仅伤了头皮还伤了脑子,是低估了成龙,成龙是演员中的战斗机,战斗机中的隐形机,他敦厚的外表下面隐藏着很高的智商。这句话居然用上了二元极致的玩法。所以那三条推特儿不是助手随意发上去的,成龙可能随意发推特吗,一个推油都不会随意的巨星,是不会随意推特的。



大家应该觉得成龙这话的句式很眼熟,想一想,就会发现其实就是前两天国资委一位领导说的:现在的老百姓是怎么啦,国有企业不赚钱你们骂,国有企业赚了钱你们还骂,要我们怎么办。明白吧,成龙确实很适合当领导了,都很凛然的句式,但其实是,拿着人民的钱的国有企业不赚钱当然骂,赚了钱却不回报人民或者干脆就不是挣钱而是抢钱,更要骂。回到菲律宾这件事,警方当然不可以不谈判就击毙绑匪,更不可以谈判十来个小时才击毙绑匪。反正我看到营救人质那一队人马时,还以为菲律宾也有民兵编制,或者是协警,后来才知道,他奶奶个腿的居然是特警。



那些悲伤的细节和那些诡异的笑容不想多谈了,有时候回忆也有罪恶感。我只是奇怪当年那个演员怎样变成一个领导的,我想他肯定想当电影局局长甚至想当文化部部长,我承认这样的揣测有些恶意,但现实告诉我们,在中国什么东西做大了,就容易叛变,不要以为内地人才会叛变,港台人也一样,成龙不再是一个演员,而是世博肉身的海宝,是奥运第六个福娃,是一切堂会剪彩的金剪刀,是维持会会长,到最后他觉得中国人确实应该管一管,而政府又管不到位,就该拿给绑匪去管一管吧。



这证明我们这个地方的某种力量足够大,大到足以把一个演员培养成为领导,其实也不一定演员才叛变,知识份子也一样,比如陈文茜前段时间说韩寒不该骂世博会,陈文茜在台湾以犀利著称,现在却当起世博唱诗班BB,可是在上海生活近三十年的韩寒还不比刚来上海三天的陈文茜更了解上海和上海的变化?这就不对,但陈文茜就是要这样说,这样说她有利益——而且这个好处必不是帮李敖炒作李戡……其实李敖也是我们那一代愤青曾经很喜欢的,因为他敢一人独抗国民党反动派和阿扁很多年,但现在他在写《阳痿美国》,在大赞内地日新月异的变化,中国内地当然有很多变化,但你独看到可喜的看不到可忧的,你不再像知识份子那般深独立去反思,而只是随便的起兴,这就不是李敖,而是李咏,不是反骨的李敖,而是看院的藏獒。



利益知识份子,利益演员,利益科学家,在这个强大的气场下,你以为他们唱的是“我们都有一个家”,其实心里是“台币人民币两手都要抓”,你以为他要做龙的传人,其实他要弄翻一船人,你看,凡是成龙代言的产品,最后都死光光,无一例外:代言小霸王,小霸王倒闭了;代言爱多VCD,爱多倒闭、老总坐牢;代言汾湟可乐,汾湟可乐没了;代言开迪汽车,全国才卖九百多辆;代言霸王洗发水,被查出霸王致癌,很快也将消失了。而成龙最新代言的另一品牌格力空调,近期也频传空调爆炸事件,成龙就是一广告毒药,代言谁谁死……这个不是我统计的,是别人,如有出入,请查。



想一想,成龙比郭德纲还要低俗的,郭胖子代言的伪劣广告没成龙多,也没一个小龙女摆在大街上,更没说过不憎恨菲律宾,可郭胖子是要下架的,成龙不会下架,成龙的电影照样放,成龙的堂会照样开,成龙的传人还要薪火相传,可见有关部门比我们都懂成龙,因为成龙比郭德纲还懂中国内地。再想一想,中国人没人管你憎恨,中国人太多自由你憎恨,中国网民乱说话你憎恨,中国记者调查霸王你憎恨,独独中国人被杀死在异国你不憎恨,这多少有点让人憎恨。



我不欣赏民族主义,也不见得就接受自由主义,我只选择人性主义,因为只是一个俗人,只搞得懂最基本的主义。上上篇“中国人为什么没安全感”被战胜了,我并不介意,因为我知道并不因为"安全感" 被战胜,中国人就有了安全感。大家知道,我们没可能改变世界,只有世界改变我们,所以我们需要偶像,因为偶像可以改变心情,带来点安全感,但是,这次人死了,成龙没有声哀鸣,只有声援菲国,这有些怪怪的,老吾老以及人之老,幼吾幼以及人之幼,如果成龙只做到妻吾妻以及人之妻,这就很让人寒心,很没安全感。

Sunday, August 22, 2010

From iPSC to induced beta-cells, iN and iCM: dedifferentiation vs direct reprogramming

The success of inducing pluripotency in primary fibroblasts and other cells with a combination of only a small number of transcription factors suggested that fully differentiated cells might change fate following similar treatments. Since the demonstration of induced pluripotent stem cells (iPSCs), at least three examples have been published where 3 cell type-specific factors were selected from a pool of 10-20 candidates that, when expressed from viral vectors, could induce beta-cells, neurons, or cardiomyocytes.

Induced beta-cells [1]: Ngn3, Pdx1, and Mafa, adenovirus injected to in vivo targets

Induced neurons (iN) [2]: Ascl1, Brn2, and Myt1l, lentivirus infecting mouse embryonic fibroblasts (MEF) or tail tip fibroblasts (TTF)

Induced cardiomyocytes (iCM) [3]: Gata4, Mef2c, and Tbx5, lentivirus infecting cardiac fibroblasts or TTF

In all 3 cases, the change of fate seemed to be via direct conversion, without passing through a progenitor cell fate before further differentiation. Like iPSC reprogramming, direct reprogramming also requires a transient supply of inducing factors. Unlike generating iPSCs, the percentage of cells getting reprogrammed is much higher in direct reprogramming, ~20% in the cases of iN and iCM vs 0.1-1% in iPSC. It is likely that a transient, inductive expression of essential factors jump-starts endogenous factors to establish cell fate specific programs; it has also been illustrated that chromatin remodeling through DNA methylation, histone modifications, etc. accompanies the direct reprogramming events.

Read the complete story and this week's official post and new product of the week, weekly promotion, etc, go here.

Saturday, August 14, 2010

Do You Know How Well Your Sunscreen Works?

Skin diseases caused by sun exposure include melanoma, basal cell carcinoma, squamous cell carcinoma, photoaging, as well as sunburn and many other conditions. According to the Skin Cancer Foundation, skin cancer is the most common type of cancer in the US. The vast majority of mutations found in melanoma, according to a 2009 study published in Nature [1], are caused by UV radiation.

Currently, commercial sunscreens are composed of physical sunblocks including zinc oxide and titanium dioxide, and chemical UV (ultraviolet lights) absorbers/filters such as octinoxate for UVB and benzophenone for UVA. The compositions of commercial sunscreen products are disclosed by the manufacturer and regulated by the health product regulatory authorities such the FDA in the US. The UV absorbers/filters are organic chemicals that absorb UV lights within a very limited range of wavelength. Consequently, a combination of different chemicals is needed to achieve "broad-spectrum" protection.

Currently the FDA required test of effectiveness of UV protection measures only UVB, which means there is no way of knowing how effective a sunscreen product is against cancer-causing UVA and damaging visible lights [2]. Even though the life style changes in recent time result in more damaging light exposure such as extended sun bathing on beach or tanning in beauty saloons, etc., only 3 new sunscreen active components (and none of new chemical class) have been introduced to the US market in more than 3 decades. There seems to be a gap between the need and the effort for developing substantially improved skin protection products.

For references, see original blog: http://allelebiotech.com/blogs/2010/08/do-you-know-how-well-your-sunscreen-works/

Wednesday, August 11, 2010

反三俗趣贴,世道不一样了,人也不好忽悠了

A couple of good ones out of thousands of posts related to the current campaing against "3 low tasts/classes" by the gov in China:

枫林晚照:

孤舟蓑笠翁,独钓寒江雪。
======================
一个老翁在小舟中独吊一个叫寒江雪的少女,证据确凿!以为将吊换成钓俺就看不出来?

hzg7:

花褪残红青杏小
=============
该禁,这鸟人在抱怨人家飞机场

mstzl518:

衣带渐宽终不悔,为伊消得人憔悴
=============
这个,又黄又凄美

Saturday, August 7, 2010

Looking back at another year

It’s that time again to thank mom, and sit down with a bottle of wine and look back at life, particularly the last year, to see whether it has been good.  Let’s see, still have a couple of true friends, even though our views of world started to differ recently, but we are still true friends as what true friends mean.  Got two boys who so far have been growing up fine, all looking good for them to become good and honorable men.  Two papers in peer-reviewed journals, more than a couple of patents granted and few more filed in the last year alone. Been taking care of dozens of employees and their families, trying to be fair and helpful with everyone involved.  

Yet nothing too dramatic has happened in recent years. Experiencing an 8.9 quake in Chengdu did not feel like anything felt back in 76 during the Tangshan earthquake.  Nothing came close to surpass the experience 11 years ago as major event in life.  It hardened me, like battle field tough, whatever bad happens now could not be as oppressive and unjust as what I had experience then, so it is actually natural to never give in, never give up.  Sometimes I can’t stop wondering what if it all ended then?  Would it be worth it?  Would there be regrets that all the things that happened in the last 11 years would not have had the chances to play out?  Would the world miss my discoveries and contributions?  Probably not, but what about the kids?  I guess hypothetical questions are pointless most of the times, because all optional assumptions are of no real consequences.  Even so, I would say yes to the “worth it” questions, seeing what has occurred to other countries since 89 and their peoples, and what is happening to the tired and the poor of my old country.  Had I died back then, I would have wished that if one day someone would put something like this to the memory of me:

“You did not bear the shame
You resisted
Sacrificing your life
For freedom, justice and honor”

A memorial to von Stauffenberg  and the entire German Resistance  

Since I didn’t become a hero, this should go to those who did sacrifice their lives.  For me, at least I can say, I did not bear the shame.  

Wednesday, August 4, 2010

MicroRNA-132 is identified as a stimulator of neovascularization

http://www.allelebiotech.com/News//index.php?mod=article&cat=RNAi&article=774

By looking for miRNAs that are upregulated by VEGF or bFGF in umbilical vein endothelial cells (HUVECs) AND an ES neovascularization model, researchers in the Cheresh group at UCSD and Moores Cancer Center and the King lab at U Michigan identified miRNA-132 as a critical switch for activating endothelial cells during angiogenesis. The result, published in Aug 1st’s Nature Medicine, is most significant in that miRNA-132 could become a target for tumor prevention given the relevance of pathological blood vessel formation to tumors.


Profiling of miRNA during pathological development has been fruitful in many cases, including one conducted by Allele Biotech’s researchers and colleagues from Nanjing Medical University where a number of miRNAs were identified with dramatic changes that are specific to several stages of adipogenesis. It is somewhat surprising that miRNA profiling has not been done in more development models or in more systematic ways. In fact, other miRNAs have been identified as stimulators of angiogenesis previously, such as miR-296, which, different from miRNA-132, is “upregulated only in growth factor-treated endothelial cells but not in the developmental angiogenesis screen using human embryonic stem cells.”

The mechanism by which miRNA-132 enhances blood vessel formation has been pinpointed to a RAS regulator. The method used for predicting miRNA targets was by using 3 algorithms for finding multiple sites in the 3’-UTR. The technique for analyzing miRNA-12 effects was mostly based on using an anti-miRNA-132 oligo. Both cell and animal models were used in this study. A type of tumor-specific integrin-targeted nanoparticle was used to deliver anti-miRNA-132 to tumor sites in mice.

Anand et al. Nature Medicine, 08/01/2010
http://www.nature.com/nm/journal/vaop/ncurrent/full/nm.2186.html

Friday, July 30, 2010

Allele’s pallet of the super star fluorescent proteins

From AlleleBlogs
http://allelebiotech.com/blogs/2010/07/alleles-pallet-of-the-super-star-fluorescent-proteins/


“Photoblog”–just some fun pictures from our notebooks.
    The brightest cyan, green fluorescent proteins, and the brightest ever FP in LanYFP!
The brightest cyan, green fluorescent proteins, and the brightest 
ever FP in LanYFP!
Ain't they pretty?
These fluorescent proteins are representatives of the growing family or high quality, new generation FPs engineered to enable experiment previously deemed impossible.
    Cells infected with lentivirus carrying mWasabi. Lentivirus carrying LanYFP will make most cells much more brighter than this.
2-3 times brighter than EGFP, no cytotoxicity detected
The mWasabi is stimulating
The brightest green fluorescent protein with excellent photostability, carried on 10e8 TU/ml high titer lentivirus.
    The LanFPs express well in bacteria.
Reminding you of icecream
The LanFPs express well in bacteria
Project planning is under way to test the cytotoxicity of lanFPs in different mammalian cell lines and in vivo with a focus on neurons.
    The FPs fold so strongly that they fluorescence even in SDS-PAGE.
Fluorescence while running in denaturing gel
Can you see the FP bands in the SDS PAGE?
    FPs in SDS PAGE–a closer look
while the gel is still running
Can you see them now?
    FPs in gel cassette over UV lights
Easier to see now than during gel running
Invincible FPs
    FPs in gel cassette under blue LED
The red FP is harder to see because of the black background
Fluorescence in SDS page under blue LED
The purified FPs can be used as “real time” protein markers.
New Product of the Week 07/26/10-08/01/10: pCHAC-mWasabi-C for expressing mWasabi fusion through retroviral vectors.
Promotion of the Week 07/26/10-08/01/10: Get 3′ TAMRA & BHQ oligo mods for $45 ea & 3′ Dabcyl mod for $20 50 nmol syn scale only/while supplies last- use dbtkrm0726

Saturday, July 24, 2010

老袁如此无耻,小爷懒得姓袁

袁世凯的侄子(侄孙?)袁瑛放炸弹谋刺老袁未果被俘。过堂时他指着坐堂的老袁大骂∶“你袁世凯胆敢称帝,小爷我就懒得姓袁”

今偶见一贴:

北佬 加帖在 猫眼看人 【凯迪网络】 http://www.kdnet.net
本人怀着沉痛的心情向禹晋永兼职客座教授的学校 - 北京大学、清华大学、人民大学、中央财经大学、中国科学院、北京化工学院 - 致哀!在广大P民的心中,你们正在死去!

遂追索禹教授新诗一首:

“心系玉树睡不着,半夜起来看微博,以为只有我一人,结果博友都没睡,都在关心玉树人,国家行动很神速,救援工作没停步,玉树幸运比汶川,我们大家都心安,祈祷难友能安魂,活人定要信乾坤,国家民众齐出手,幸福生活样样有!”

赏析(转):首先,此诗总长十三句,对转韵的运用放浪果敢,颇有唐乐府之风;“心系玉树睡不着,半夜起来看微博”一句、押的是方言古音,朴拙大气;“以为只有我一人,结果博友都没睡”,构想新意,不落窠臼;“玉树幸运比汶川,我们大家都心安”、“国家民众齐出手,幸福生活样样有”句,颇有讽世辛辣况味;单从 “活人定要信乾坤”句,足可说明诗人知敬畏,安天命,绝不妄自尊大说“胜乾坤”等;最后,诗人的悲愤在“祈祷难友能还魂”句达到了高潮,难友一词,无居高临下态,指你我无论生死,不过是难兄难弟,只争来早与来迟,祈祷“能还魂”,则更从态度转向了行动:“能还魂”意指“来索命”,如何索命,又索谁之命?故此诗似歌功颂德而内有褒贬,皮里阳秋,微言大义也,实为近年来少见的虎狼之作


北大有如此人物,我懒得说是北大人。

Saturday, July 10, 2010

华夏快递 : 格丘山:重读爱因斯坦的“我的信仰”

没有一篇文章像爱因斯坦的"我的信仰"令我这样感动,在"我的信仰"这篇短文中,爱因斯坦以一个科学家的诚实和简洁说明了他的人生观,宗教观,政治观。

对于人生,爱因斯坦说:

"要追究一个人自己或一切生物生存的意义或目的,从客观的观点看来,我总觉得是愚蠢可笑的。可是每个人都有一定的理想,这种理想决定着他的努力和判断的方向。就在这个意义上,我从来不把安逸和享乐看作是生活目的本身""这种伦理基础,我叫它猪栏的理想。照亮我的道路,并且不断地给我新的勇气去愉快地正视生活的理想,是善、美和真。要是没有志同道合者之间的亲切感情,要不是全神贯注于客观世界""那个在艺术和科学工作领域里永远达不到的对象,那末在我看来,生活就会是空虚的。人们所努力追求的庸俗的目标""财产、虚荣、奢侈的生活""我总觉得都是可鄙的。"

对于政治,爱因斯坦说:

"我的政治理想是民主主义。让每一个人都作为个人而受到尊重,而不让任何人成为崇拜的偶像。我自己受到了人们过分的赞扬和尊敬,这不是由于我自己的过错,也不是由于我自己的功劳,而实在是一种命运的嘲弄。其原因大概在于人们有一种愿望,想理解我以自己的微薄绵力通过不断的斗争所获得的少数几个观念,而这种愿望有很多人却未能实现。我完全明白,一个组织要实现它的目的,就必须有一个人去思考,去指挥,并且全面担负起责任来。但是被领导的人不应当受到强迫,他们必须有可能来选择自己的领袖。在我看来,强迫的专制制度很快就会腐化堕落。因为暴力所招引来的总是一些品德低劣的人,而且我相信,天才的暴君总是由无赖来继承,这是一条千古不易的规律。"",在人生的丰富多彩的表演中,我觉得真正可贵的,不是政治上的国家,而是有创造性的,有感情的个人,是人格;只有个人才能创造出高尚的和卓越的东西,而群众本身在思想上总是迟钝的,在感觉上也总是迟钝的。"

对于战争,爱因斯坦说:

"我想起了群众生活中最坏的一种表现,那就是使我厌恶的军事制度。一个人能够洋洋得意地随着军乐队在四列纵队里行进,单凭这一点就足以使我对他轻视。他所以长了一个大脑,只是出于误会;单单一根脊髓就可满足他的全部需要了。文明国家的这种罪恶的渊薮,应当尽快加以消灭。由命令而产生的勇敢行为,毫无意义的暴行,以及在爱国主义名义下一切可恶的胡闹,所有这些都使我深恶痛绝,在我看来,战争是多么卑鄙、下流!我宁愿被千刀万剐,也不愿参预这种可憎的勾当。尽管如此,我对人类的评价还是十分高的,我相信,要是人民的健康感情没有被那些通过学校和报纸而起作用的商业利益和政治利益蓄意进行败坏,那末战争这个妖魔早就该绝迹了。"

对于宗教,爱因斯坦说:

"我们所能有的最美好的经验是奥秘的经验。它是坚守在真正艺术和真正科学发源地上的基本感情。谁要是体验不到它,谁要是不再有好奇心也不再有惊讶的感觉,他就无异于行尸走肉,他的眼睛是迷糊不清的。就是这样奥秘的经验虽然掺杂着恐怖产生了宗教。我们认识到有某种为我们所不能洞察的东西存在,感觉到那种只能以其最原始的形式为我们感受到的最深奥的理性和最灿烂的美正是这种认识和这种情感构成了真正的宗教感情;在这个意义上,而且也只是在这个意义上,我才是一个具有深挚的宗教感情的人。我无法想象一个会对自己的创造物加以赏罚的上帝,也无法想象它会有像在我们自己身上所体验到的那样一种意志。我不能也不愿去想象一个人在肉体死亡以后还会继续活着;让那些脆弱的灵魂,由于恐惧或者由于可笑的唯我论,去拿这种思想当宝贝吧!我自己只求满足于生命永恒的奥秘,满足于觉察现存世界的神奇的结构,窥见它的一鳞半爪,并且以诚挚的努力去领悟在自然界中显示出来的那个理性的一部分,即使只是其极小的一部分,我也就心满意足了。"

我想诸位也许能够比我从这些话中吸收到更多的真谛,最后让我抄录爱因斯坦在悼念玛丽·居里夫人时说的话与大家共勉,"居里夫人的品德和热忱,哪怕只要有一小部分存在于欧洲的知识分子中间,欧洲就会面临一个比较光明的未来"。

我想,对中国也一样。

Thursday, July 8, 2010

Entrepreneurs Roundtable in San Diego: How to Get Seed Funding

Wednesday July 28th at 5:00 p.m.



Johnson and Johnson Auditorium

3210 Merryfield Row, San Diego, CA

Hear from successful entrepreneurs
Make connections
Take your idea or company to the next stage

Program

5:00 - 6:00 p.m. Networking & Refreshments

6:00 -7:00 p.m. Roundtable & Discussion: How to Get Seed Funding

Confirmed Speakers

Court Turner (Avalon Ventures)

Raj Krishnan (Founder, Biological Dynamics)

Malcolm Bohm (Founder, Trialytics)

Moderator: Doug Lappi

7:00 -7:15 p.m. 30 Second Pitch Segment

(pitch your company/service in 30 sec.)

Please send a short pitch proposal to pitch@sdentrepreneurs.org by July 26th for consideration

Please register via

http://seedfundingjuly28th.eventbrite.com

This event is free to the public. Refreshments provided. Donations and sponsorship are welcome.

The event is sponsored by the San Diego Entrepreneurs Exchange (www.sdentrepreneurs.org). The SDEE was founded by local entrepreneurs in order to provide a voice for the early stage technology startup, to encourage new entrepreneurs, and to sponsor networking and educational events to help develop the skills necessary to bring funding and business to the San Diego area.

Allele News Release, editor JW http://www.allelebiotech.com/News//index.php?mod=article&cat=OtherNewsRelease&article=692

Saturday, July 3, 2010

Updates on dealing with banks

In earlier blogs I wrote how banks behaved in relevance to both of my business and personal accounts. I paid much attention as part of a learning experience because banks represent the type of business that my company deals with at the highest frequency among all commercial services. Previously I complained about CapitalOne for not making efforts to understand a situation that unfairly resulted in increase of interest rate. They later changed that and now gave my company the lowest rates among all accounts. I thought for a well that Discover was friendly and reasonable but it turned out that they increased our rate without notice to above 25% after agreeing on phone to reverse a rate increase, just the opposite to CapitalOne, and of course they are history, I closed the Discovery account never again will I have them for business funding needs.

Advanta's underwriting bank got into some trouble during last year's turmoil and shut down our account, and it was gone from our list of credit services for good as well after I paid it off quickly. What's funny is that for some reason Citi bank kept sending monthly statements to my home address, maybe thinking that linking my business account closer to myself would make it safer? After repeated problems of miss-delivered (to my old address) and late payment issues, we seemed to have cleared it all up by a bunch of phone calls and finally using our business address. However, just last week they decided to close our account based on these payment issues. It struck me that severe miscalculations that these large banks have through their staff that make judgment calls on customers' credit worthiness still exist after they run themselves to the ground, had to use your and my tax money for a bail out that almost tore apart our nation, cost me big losses on their stocks, now they tell us good customers that we do not deserve their services. The lesson learned, a company has to have a system for operations, but that system must have a common sense check point in place.

I am not against just citi bank, of which I am a share holder and I hope they would do well, for being quite stupid while probably feeling kinda smart on making such moves through their customer accounts; or BOA who completely messed up my personal account for more than 20 years and lost me as a customer for ever; or Union Bank who on one hand tried to lure my company's business, on the other hand had a bunch of really bad business people working on my issues. I closed BOA, then Union bank checking, loan, and cutting down the last dealing with them.

Just a blog for other peeps or biz to learn something from my first-hand dealing with these banks who contributed handily and heavily to troubles many of us are experiencing now. As a businessman, I will continue to find the best service, and take risks of getting funding to continue hire new people and helping my employees through the good time and the tough times. As an executive, I learn from other business and try to avoid doing things that are unreasonable, unethical, or plain stupid. As a scientist, I try to put more reasoning and logic in figuring out human factors in all these events.

Thursday, July 1, 2010

Allele Custom Services for Drug Screening Companies

http://allelebiotech.com/blogs/2010/06/allele-custom-services-for-drug-screening-companies/
Many target discovery and validation programs can benefit from RNA interference, fluorescent proteins, stem cells, and viral delivery systems. However, applications of these technologies require special reagents and laboratory know-how. Even when available, many generic reagent kits are not tailored for your particular needs in screening or validation.

At Allele, we accelerate your discovery efforts with custom RNAi screening, fluorescence based assays, and cell model development services.

1) Our RNAi platform, based on our patented shRNA/miRNA technologies, use DNA linear template, plasmid, lentivirus, retrovirus, or baculovirus vectors that prompt cells to endogenously express RNAi. As a result, our screens offer advantages over synthetic siRNAs:
• Higher levels of consistency
• Greater delivery and gene silencing efficiencies
• Accessibility to difficult-to-transfect cells, including primary cells
• Potential for inducible RNAi expression
• More persistent silencing with shRNA under Allele’s own IP–you may not need to license siRNA patents!

2) Fluorescent proteins (FPs), which can span the entire visual spectrum, have become some of the most widely used genetically encoded tags. Genes encoding FPs alone or as fusions to a protein of interest may be introduced to cells by a number of different methods, including simple plasmid transfection or viral transduction. Allele Biotech is one of a few companies that develop and improve FPs through fundamental research. We have so far achieved:
• The brightest cyan and green FPs, true monomers for minimum artifact or cytotoxicity
• The brightest yellow and red FPs from lancelet, only FPs from vertebrate
• mTFP1 as the best FRET donor by 3 independent reports
• Photoconvertible FPs for super imaging or kinetic labeling
• Delivery on plasmid, retrovirus, lentivirus, baculovirus vectors

3) As a major advancement in the stem cell field, it has recently been shown that mouse and human differentiated cells may be reprogrammed into stem-like, pluripotent cells by the introduction of defined transcription factors. These induced stem cells (iPSCs) provide unprecedented resources of cells of different differentiation stages for functional testing and drug screening. Allele Biotech develops and provides state-of-the-art reagents in convenient forms for iPSC production
• iPS factors carried on lentivirus, retrovirus, baculovirus for different cell types
• Availability in combination with fluorescent proteins under own IP, and drug resistant genes
• 4-in-1 or 2-in-1 effective use of iPS factors on one viral vector
• Feeder cells of human origin expressing factors essential for stem cell culturing

4) Introduction of protein factors, miRNA, promoter-reporter, and virtually any other genetic element of interest via the most efficient viral packaging systems.
• Introducing protein-FP fusion, promoter-FP reporter, photoactivatable factors for cell-based assays
• Introducing critical factors for cell immortalization
• Episomal or integrated expression using baculoviral vectors
• High throughput, systematic expression of whole class of molecules in any type of cell
• High titer viral packaging at low cost for delivery to animal tissues

In addition, the Allele team can provide custom-designed assays that can be used for assaying enzyme activities in almost any pathway, such as the EGF pathway, TNF response/apoptosis pathway, nuclear receptors, etc. We utilize technically advanced methods to provide our partners with advantages over alternative methods or other services.

New Product of the Week 06-28-10 to 07-03-10: Eco-friendly mammalian tissue culture plates, 40% less plastic to the environment, 40% less cost to your budget, contact our sales rep today for quotes and details.

Promotion of the Week 06-28-10 to 07-03-10: Oct3/4 iPS lentivirus with RFP as marker, new to the market, this week only all kits containing Oct3/4-RFP same price as the original, non-RFP versions, save ~$50!

Thursday, June 24, 2010

Brightest Ever Fluorescent Protein

http://allelebiotech.com/blogs/2010/06/brightest-ever-fluorescent-protein-2/

LanYFP, identified from lancelet (also known as amphioxus, e.g. Branchiostoma floridae), has been found to have the following properties:

Excitation 513nm
Emission 524nm
Quantum yield 0.95
Extinction coefficient 150,000
pKa ~3.5
Salt insensitive 0-500mM NaCl

LanYFP has a brightness of 143! For comparison, the brightness of the previously known brightest FPs is 95 for tdTomato, and 34 for commonly used EGFP.

Allele already has been exclusively providing the brightest cyan FP in mTFP1 (brightness of 54); and the brightest green FP in mWasabi (brightness of 56). The confirmation of LanYFP as the brightest ever FP is a major milestone of Allele’s research and development efforts in the fluorescent protein field. We are currently monomerizing LanYFP and another lancelet protein, LanRFP. Once completed, the new proteins should definitely be the FPs of choice for in vivo imaging and FRET with unprecedented utilities.

Saturday, June 19, 2010

From Nature Biotechnology: SBIR grants wax

Awards under the Small Business Innovation Research (SBIR) program have just been given a boost. As of March 30, the cap for SBIR phase I awards has risen from $100,000 to $150,000, and for phase II awards from $750,000 to $1,000,000. The increases are intended to take account of inflation since 1992 when the threshold amounts were last set by Congress. "This will have an important positive impact at a critical [juncture] in the aftermath of the nation's great recession," says Simcha Jong, university lecturer in management science and innovation at University College London. Jong says that, historically, the SBIR program helped forge links between university science and industry and, at this pivotal time, could help kick-start the US job engine. The Senate has passed a bill to extend the SBIR and related Small Business Technology Transfer through July 31 (Nat. Biotechnol. 27, 1065-1066, 2009). Even more generous than SBIR grants are the new Small Business Helping Investigators to Fuel the Translation of Scientific Discoveries (SHIFT) awards launched on March 5 by the US Department of Health and Human Services. These awards, aimed at fostering translational research, offer companies up to $2.65 million over five years. "The main point is to encourage current academic researchers to apply, and use it to move to biotech," says Jiwu Wang, president and CEO of Allele, a San Diego-based company that has taken products to market with SBIR support. "It is a great idea."

Emma Dorey, Nature Biotechnology

Sunday, June 13, 2010

Oct3/4 Promoter Blocks Cancer Stem Cells from Differentiation—Potential for Stably Supplying CSCs

Cancer stem cells (CSCs) are thought to be a group of cells within tumor tissues that are resistant to standard chemotherapy and the main cause for relapse. Typically identified by cell surface markers (e.g. CD44, CD24), CSC cells belong to a minority population among cancer cells, which are difficult to obtain or maintain. There have been several NIH programs soliciting novel methods of isolating CSCs for functional studies and screen for therapies.



Sajithlal et al. reported in Stem Cells that by introducing an Oct3/4 promoter-GFP construct in an attempt to follow CSCs among MCF7 breast cancer cells, they unexpectedly found out that this construct blocked differentiation of CSCs and maintained their signature cell surface marker expression profile. It is not easy to explain how a promoter (assuming the expressed GFP has nothing to do with the effects of this construct) would block differentiation, however, the authors did speculate that it could be some ORFs or small RNAs encoded within the 4 kb promoter region, or its competition with endogenous promoter. They went on to test the same construct in several other breast cancer cell lines and found similar results. It remains unclear whether it would be true in other cancer types, isolated cancer cells instead of cell lines, or if other stem cell specific promoters, such as Nanog promoter, would have similar effects.



For those who are interested in CSCs, this new finding should be of great interest for theoretical as well as practical reasons.


STEM CELLS 2010;28:1008–1018 www.StemCells.com, photo courtesy


AlleleNews JW



Related product from Allele Biotech iPS product line; Pluripotency reporter lentivirus, ready-to-use , e.g. Validated Oct-4 Promoter Reporter Lentiviral Particles, 5 vials ABP-SC-ROCT4G5 $450.00.

Monday, June 7, 2010

Introducing Product-on-Demand Biological Research Reagents

http://allelebiotech.com/blogs/2010/06/introducing-product-on-demand-not-going-to-be-called-ipod-biological-research-reagents/

The general order of operations in the bioreagent industry begins with a developer observing or forecasting a need and developing a product. The supplier then supplies that product to customers by showing that the product will suit their existing needs. An alternative order in our industry is after a new discovery in the form an enzyme reaction mechanism, affinity binding, or biological system is made in lab, someone realizes that discovery could be made into a product. If the idea is picked up by a commercial R&D team, the underlining mechanisms of the discovery are then exploited for particular use and reagents or kits will be built around it. The new products are introduced to the market by convincing potential users that they will make their research better, cheaper, or faster.

From a supplier’s point of view, if the current processes for developing new products have been working, what’s the incentive to change? From a researcher’s point of view, well, do they have any other choices? If something is not commercially available, someone will just make it in the lab if they need it. Some of us still remember the days when a graduate student needed to make his own restriction enzyme because NEB didn’t sell it. However, there is a disconnect between how much new knowledge is being gained every single day in tens of thousands of labs and how small a portion of that knowledge pool is being turned into more powerful tools to make the next round of research easier and more cost-effective. For instance, when an important gene’s promoter is recently defined by a functional study in 293T cells, how soon do you expect to test the signals that influence transcription from that promoter in the primary cells you are working on? Wouldn’t it be nice if you could simply buy a vector that will express a promoter-driven reporter ready to be introduced into the primary cells in your lab instead of having a graduate student design, construct, learn and try to make a lentiviral vector in the next few months?

And yes, there is the route called custom projects provided by a few bioreagent companies. The prices are often inhibiting for the reasons that the price needs to cover for labor on industry pay scale, materials, indirect, and profit. Additionally, since the service provider does not take ownership of the product, the work of researching the relevant pathways and making construct designs is left to the user.

There is a better way. A company can plan product groups, lines, and packages based solely on the demonstrated importance of a system such as signal pathway or a family of molecules like miRNA. The plan can project to use the most advanced technologies, even accompanied with full product descriptions and vector maps. However, it would be a great waste of money and material if nobody would ever need it, right? One way of dealing with the initial cost is that we make the first kit upon the first order. The customer that places the first order of a new product will get a deep discount off the shelf-product price on what used to be a custom project. They might even have the opportunity to provide input on the product design prior to production. From a supplier side, we will benefit by having an opportunity to initiate a new product without major investment, which in turn would keep our overall prices low for such innovative and advanced products.

This model should help speed up the commercial application of any new biological findings, lower the cost and price of bioreagent products, and encourage interaction between researchers who normally do not work with each other to produce better products for increasing the efficiency of research.

Discount of the week 060110-060710: Any virus packaging project initiated this week gets additional 10% discount that can be used with first time discount and other pricing advantages. http://www.allelebiotech.com/allele3/Services_Lentiviral_Retroviral_Packaging.php

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