a****d
发帖数: 1919 | 1 个人经验， lentivirus （ubiqutin promoter） 感染的细胞，12小时后可以看到eGFP
表达。 CAG-CMV promoter based lenti也能很快看到外源基因表达。
我培养的原代细胞，D0用lenti处理，D3 固定染色，没有问题。 |
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d*****r
发帖数: 45 | 2 不是大牛
pcDNA 完全可以，你也可以试试vectors with CAG promoter，理论上表达更强.
另外，你可能要先make sure HEK293 里有没有内源性的potassium currents ， 会不
会干扰hERG currents. |
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m********2
发帖数: 317 | 3 是个好工具，但是如果想用人的细胞就还需要cas9病毒了
这个小鼠也有个问题就是，不知道CAG promoter是不是universe表达的
knockout |
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L****T
发帖数: 169 | 4 Most people in this field are now using piggybac vector with CAG promoter |
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f*******e
发帖数: 354 | 6 不要用mscv，cmv这些常规retrovirus的promoter，很快会被silence，可以用plenti，
psin，phage等，ef1a, cag, pgk可以坚持一段时间，但是也存在被silence等现象.转
质粒总体比较困难，病毒是最高效的方法。在细胞传代的同时加lentivirus，
polybrene，然后spin半小时，效率会很高. rock inhibitor记得加，不然细胞会死光
光.病毒含血清的话几个小时后就可以换掉，如果要drug selection，过夜也没事，第
二天可以继续加rock inhibitor，浓度可以降到5um。
good luck！
: 非常感谢！我好好读下pMSCV retroviruses的manual。你觉得用pCDH是因为这些
: transgene被silenced吗？还是有其他的原因？非常非常感谢！
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s*****g
发帖数: 3693 | 7 “The trinucleotide contexts characteristic of AA exposure included a
prominent peak at 5′-CTG-3′ (5′-CAG-3′ on the complementary strand)”
马兜铃素引起的T:A >A:T变化对周边序列是有偏向的，所以才叫signature. 你引用的
nature genetics文章没有提及任何的序列偏向。
至于你说的第二篇文章，需要高剂量是因为小鼠在喂食之后24小时就杀了取样 “
Groups of three rats received a single oral dose of 0, 5 or 30 mg/kg body
weight of AA in 1% NaHCO3 solution, respectively. Rats were sacrificed 24 h
after the AA dosing by decapitation. Kidney and liver samples were collected
”，你猜测的肝代谢没有证据。24小时和... 阅读全帖 |
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y******m
发帖数: 143 | 8 这是发表在STM2013年的文章，实际上发现了HBV引起的突变和AA signature类似，但是
他们把它归为可能由AA引起的。最精彩的是最后一段，因为这些病人的AA exposure的
信息没法获取，所以推测是由AA引起的。这个还能没牙齿点嘛！！！仔细看了下，2017
年的样本也许就是2013年的样本，这个HBV样本AA signature分布和2017年的图2是基本
一样的。
We detected 11 cases exhibiting strong AA (or AA-like) mutational signatures
using the following criteria: (i) A > T is the most frequent mutation
category or there are ≥10 exonic A:T to T:A transversions; (ii) ≥33% of
the A:T to T:A transversions are associated with a C|TAG sequence context;
and (iii) a s... 阅读全帖 |
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s*****g
发帖数: 3693 | 9 “The trinucleotide contexts characteristic of AA exposure included a
prominent peak at 5′-CTG-3′ (5′-CAG-3′ on the complementary strand)”
马兜铃素引起的T:A >A:T变化对周边序列是有偏向的，所以才叫signature. 你引用的
nature genetics文章没有提及任何的序列偏向。
至于你说的第二篇文章，需要高剂量是因为小鼠在喂食之后24小时就杀了取样 “
Groups of three rats received a single oral dose of 0, 5 or 30 mg/kg body
weight of AA in 1% NaHCO3 solution, respectively. Rats were sacrificed 24 h
after the AA dosing by decapitation. Kidney and liver samples were collected
”，你猜测的肝代谢没有证据。24小时和... 阅读全帖 |
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y******m
发帖数: 143 | 10 这是发表在STM2013年的文章，实际上发现了HBV引起的突变和AA signature类似，但是
他们把它归为可能由AA引起的。最精彩的是最后一段，因为这些病人的AA exposure的
信息没法获取，所以推测是由AA引起的。这个还能没牙齿点嘛！！！仔细看了下，2017
年的样本也许就是2013年的样本，这个HBV样本AA signature分布和2017年的图2是基本
一样的。
We detected 11 cases exhibiting strong AA (or AA-like) mutational signatures
using the following criteria: (i) A > T is the most frequent mutation
category or there are ≥10 exonic A:T to T:A transversions; (ii) ≥33% of
the A:T to T:A transversions are associated with a C|TAG sequence context;
and (iii) a s... 阅读全帖 |
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s********p
发帖数: 1319 | 12 Usmle大虾说得非常正确.
国内中成药自己长期以来定位就不正确,他们不但要走向治疗药的行列,还冲进急诊行列
.动不动就和民族文化,爱国主义等牵扯在一起.
这些乱七八糟的东西,不过就是加点西药成份而已.这样,实际上就是把一点传统文化的
东西扼杀了.
对危险人群,ST-T动态改变加症状具有确诊价值.但ACS很多情况下都是紧急情况,比较难
得到具有诊价值的两份对比的心电图.在急诊情况下,ACS患者的心电图不可能没有改变,
但有时这些改变都是非特异性的,而且要和PE等鉴别,目前最好的办法:立即查cTnT或
cTnI,症状加生化标记物阳性,立即到导管室.......
对危险人群,症状不典型加非特异性ST-T改变,即使有以往的ECG对比,为了给患者一个比
较明确的答案,还是要做运动负荷试验,如果阳性,收入院择期CAG......,如果阴性,有危
险因素,则药物干预危险因素.
包括血脂分项这些东西,如果是有两次结果对比,价值要大得多.
ST-T动态改变对一种人群几乎没有什么诊断价值,而且一些刚到心内科轮转的医生要上
当:正常的中青年女性_____这么说吧,45-50岁以前的女性吧,她们中有一部分人 |
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l********y
发帖数: 2295 | 13 for step1:
trinucleotide repeat expansion diseases:
My Ex is Hungry on Fri.
My (大家都Call Tall Girl, CTG)=Myotonic dystrophy
Ex (比较肉麻的Call Ge Ge, CGG)=fragile X syndrome
Hungry (点了个鸡蛋Call An Egg, CAG)=Huntington's disease
Fri (大家AA制的，Go AA, GAA)=Friedreich's ataxia
呵呵，编口诀的好处就是记得忒牢，到现在随便什么人问我哪个病是哪个repeat，我随
口就答，人家还以为我学的多好呢，呵呵。 |
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l********y
发帖数: 2295 | 14 【 以下文字转载自 Pre_Resident_Club 俱乐部 】
发信人: lovelypony (lovelypony), 信区: Pre_Resident_Club
标 题: steps学习的口诀
发信站: BBS 未名空间站 (Fri Oct 22 21:56:46 2010, 美东)
响应老刀的号召，冒个小泡。有很多难记的东西，编过一些口诀，先贡献一个，回头有
空了再慢慢添。大家有好的口诀也都来分享，这样大家都省时省力。
先贡献一个pony比较经典的口诀，认识pony的都会的，呵呵。
for step1:
trinucleotide repeat expansion diseases:
My Ex is Hungry on Fri.
My (大家都Call Tall Girl, CTG)=Myotonic dystrophy
Ex (比较肉麻的Call Ge Ge, CGG)=fragile X syndrome
Hungry (点了个鸡蛋Call An Egg, CAG)=Huntington's disease
Fri (大家AA制的，Go AA, GAA)=Fried... 阅读全帖 |
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s********p
发帖数: 1319 | 15 各大虾，蒙几位XDJM们看重，码了这帖。SCD是个比较大的问题，我天天表示要学逆风
同学的文风，但不得要领，当然，圣诞节快来了，也想从版主那里，那个那个那个——
到商场时更High些，最后还是码得太长，另起一帖……
心脏性猝死（Sudden Cardiac Death），是指心脏原因所致的、起病后1小时、或2小时
或24小时内的死亡（有争议）。主要原因是VT/VF（主要）、或心脏停博或电机分离，
血液动力学迅速归于零——大脑缺乏有效的灌注，患者意识突然丧失——大脑只能耐受
2-3分钟的缺血、缺氧！！！
患者神智丧失，大动脉无搏动（10秒种内完成判断）：
——立即当胸一拳（扣击复律），无现成的除颤器，立即持续心脏按压，进入初级生命
支持（BLS），同时呼救。不强调口对口人工呼吸，但注意开放气道，面罩给氧等还是
致关重要的；
——有现成的除颤器，除颤——目击者除颤！！
尽管美国的消防队员、警察都要经过心肺复苏培训后才能上岗，尽管各大国际机场都摆
好除颤器强调目击者除颤，但心脏性猝死问题，多年来没有重大改观，仍然是全球面临
的一个重大公共健康问题——因为，SCD是个突发事件；也许就是个首发事... 阅读全帖 |
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d**o
发帖数: 618 | 16 Hi springstep, thank you very much for your prompt and detailed answer! Now
I understand this much much better. Indeed, I ignored that baseline ST
elevation would make EKG useless for angina pectoris. Also, stress test has
the value of being a functional test, while CAG is an anatomic test. For the
old lady patient I described, maybe she indeed does not have CAD, as I
recall, she described chest *pain* rather than *discomfort*, and it lasted
long time (certainly more than 10 minutes). Indeed it ... 阅读全帖 |
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s********p
发帖数: 1319 | 17 刚给J of ECK 审了篇稿：急诊室：患者胸部不适，V1-V3 ST-T改变，在没有TnT和TnI
的前提下，拉到Cath，CAG阴性,药物诱导的Brugada Syndrome样改变，“要吸取教训”
，——这种教训多了！
当即下Turn Down——并在审稿意见中对其严厉训斥。
——不管是美国这面那个心脏中心和医院急诊室来的，告诉Editor:问对方，敢不敢
Defense? |
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u***e
发帖数: 611 | 18 不是跟你抬杠啊，我也不是心内科的，只是觉得有点奇怪。在国内，如果急诊室来了个
70岁男性胸痛，有DM,HTN病史，EKG有多导联的2mm STD，刚好他一个星期前有个EKG是
正常的。serial TNT是阴性的，难道你们会让他先去做stress test?
这个是high risk for unstable angina,在这里的处理肯定是ASA,plavix,beta-
blocker,lovenox,该给的药都给了然后cardiologist会schedule cath. 一般是48小时
内，但也有人说越早越好的。
如果有这里的cardiologist澄清一下就好了。
TnI
取教训”
得有必要，可以Defense.
Rejected了——急诊ST-T改变 + 心前区不适，可以数出不下20种情况，CAG是最潇洒的
确定诊办法，PCI
上，为什么要先强调无创伤检查？为什么要强调cTnT,cTnI这些标记物？
和共同看法，不具备法律效力。 |
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b*******s
发帖数: 954 | 19 第一道题，在elora 以及jbh2009的回答上，再补充一下
是Fragile X。
(1) X-link的疾病，现在的看法是x-dominant或者x-recessive的提法签妥当(请看
uptodate或者其他资源）
(2) 对Fragile X来说，5'-UTR的CGG有expansion.
Normal: CGG repeats < 45
Grey zone: 45 < CGG repeats < 55
Premutation: 55 < CGG repeats < 200
Premutation由于有较长的5'-UTR转录出的RNA, 这些RNA可能会特别稳定，作为sink,
和一些重要的蛋白作用，影响这些蛋白的功能。
Full mutation: 200 < CGG repeats
Full mutation的5'UTR被methylated, FXMR蛋白无法表达
Premutation和full mutation的重要差别是premutation仍然可以表达FXMR蛋白。所以
premutation和full mutation的症状非常不一样。premutation... 阅读全帖 |
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g***e
发帖数: 577 | 20 【 以下文字转载自 Stock 讨论区 】
发信人: geome (老网友), 信区: Stock
标 题: QTS_Research Updates
发信站: BBS 未名空间站 (Tue Mar 7 18:00:08 2017, 美东)
Hi,
Continue on the project, I would like to post the preliminary result of
backtest for all SP500 underlyers using Moving Average Stategy for the last
10 years: (ie, buy/sell signal by crossing of 10day and 25day average ).
Sorry for the messy format - you can get a cleaner download of the csv file
in-
http://www.icanbeatmarket.com/research_reports.html
ticker return ... 阅读全帖 |
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G*********6
发帖数: 39 | 21 心脏超声，主要看的是心脏的血流和结构。我想，作为常规，都应该做了。
但你妈妈现在的主要问题，是LCA那个病变。 LCA，是供应左心室侧壁和后壁的，声学
窗口不好！
即使心脏超声发现室壁运动障碍、运动不协调，有助于冠心病的诊断，但你妈妈现在的
问题，不是诊断问题，而是CAG明确下一步该怎么办的问题——需不需要进一步干预？
如何站在你妈妈的角度，选择一个最佳的“费用-效益”比治疗？
即使超声发现室壁运动障碍、运动不协调，目前都还是“冬眠心肌”导致的——如果有
这些情况，根本的手段，还是血运重建，可以恢复的！
你妈妈的情况，如果需要干预，目前的干预，得益是最大的！
记住：你妈妈那医生开的药我看了，没有乱开乱七八糟的保健药，这就是好医生——他
说的是对的，听他的！！——女性更年期后，一旦有冠心病，进展比较快，现在的积极
干预，作为非常大——听经管医生的！！！ |
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s********p
发帖数: 1319 | 22 来自主题: Medicalpractice版 - 心脏支架手术 赞南大！
国内的主要问题，就是医疗卫生资源和水平很不平衡。
好多地方模糊“冠状动脉粥样硬化”和“冠状动脉粥样硬化性心脏病”的诊断。好多医
院都不开展运动负荷试验，“胸部不适”，动不动就是“CTA”，CTA发现70%或者以上
狭窄，就CAG + 支架。如果CTA的狭窄程度不达标，就是压力导丝等东西，总是往支架
方向引。
支架过度问题，在国内比较严重。
用？ |
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w*******y
发帖数: 60932 | 23 Until 7/31 you can get 25% off pre-owned/used games at Gamestop that are 19.
99 and up with the following code:
25USED
This should stack with SAVER to get free shipping when you spend 25 or more.
Enjoy!
Link:
http://www.gamestop.com/
credit: CAG
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w*******y
发帖数: 60932 | 25 Go to Mountain Dew's facebook page:
http://www.facebook.com/mountaindew
, click the Like button, then click the 1 Million tab and enter your info
to get a free wristband. 8-12 weeks for delivery.
to PepsiDriver @ CAG for the heads up
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w*******y
发帖数: 60932 | 26 Link:
http://www.cheapassgamer.com/forums/showthread.php?t=264039
Like the title says, KMart will be giving a $20 game credit with the
purchase of SC2 starting 7/27 and lasting until 7/31. Maybe Amazon or
Walmart will price match.
Credit over to CAG and Sears/KMart's official representative (SHC-Gamer)
over there.
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w*******y
发帖数: 60932 | 30 In the Xbox 360 section look for a sign advertising the $399 Halo Reach
bundle. In small print on the right hand side it says "Also Available Halo
Reach 12 month card $19.99" I was able to get one for this price (after 2
managers). YMMV as stores may have pulled the signs or posted correction
notices. Credit to the pics on CAG.
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w*******y
发帖数: 60932 | 32 From Cag, Sale ends 10/04, as far as I understand all come with free
shipping for nonmembers:
360:
James Cameron's Avatar: The Game $7.99
Dark Void $9.99
Prince of Persia $9.99
Aliens vs. Predator $12.99
Darksiders $12.99
Need for Speed: Shift $12.99
Prince of Persia: The Forgotten Sands $12.99
Assassin's Creed 2 $14.99
BioShock 2 $14.99
Dante's Inferno $14.99
Alpha Protocol $14.99
Mass Effect 2 $17.99
Final Fantasy XIII $17.99
Grand Theft Auto: Episodes from Liberty City $17.99
Left 4 Dead 2 $1 |
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w*******y
发帖数: 60932 | 36 Credit goes to CheapyD on CAG forums
Link:
http://www.play-asia.com/paOS-13-71-dw-49-en-70-3f33.html
According the message posts, no issues playing on xbox live and according to
the item description, voices are in english. You will likely have issues
reselling on amazon or other places, as you can see the cover is not the
standard US cover.
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w*******y
发帖数: 60932 | 37 Wbshop.com has a sale on a few box sets. The most noteworthy (to me) is The
Ultimate Matrix collection on Blu-ray for $39.99.
Use the code ANNMES to take off another $10 bringing it to $29.99. Shipping
is $5, OR better yet if you find a cheap filler and use the code FREESHIP,
shipping is free.
Ultimate Matrix Collection:
http://www.wbshop.com/Ultimate-Matrix-Collection-The/1000025732,default,pd.html?cgid=holidaysets
Here are some other deals worth mentioning. Note: All of these prices are
after ... 阅读全帖 |
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w*******y
发帖数: 60932 | 38 According to the Kmart Ad leaked on CAG, NBA Jam for the Xbox 360 and
Playstation 3 will be on sale for $29.99, starting Sunday (12/5). This is
perfect for me since I got several $15 coupons from this week's Kmart deals
so I'll be picking it up for $15 + tax.
No link at this point, only the Wii version is showing up on their site.
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w*******y
发帖数: 60932 | 39 Thanks chelosera91 from CAG
Tank Battle for $0.99 on Playstation Network
Great Strategy Game!
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w*******y
发帖数: 60932 | 42 This deal is for UK, but you can get a free copy of Burnout Ultimate through
a UK proxy.
Use the UK proxy: Link:
http://www.docoja.com/blue
Insert this link into the proxy: Link:
http://www.hotukdeals.com/freebies/b...e-box-p/834553
Then click on the burnout paradise picture on the right that says "Go to
deal". And that should bring you to the checkout in EA store!
Got it from CAG: THEBIGHAM
Be patient, you will get it for free. Also, download EADM separately, and
use your EA account and this ga... 阅读全帖 |
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w*******y
发帖数: 60932 | 43 Supprised this is not posted here, or maybe the search engine isn't working.
Either way, here it is...
This deal is for UK, but you can get a free copy of Burnout Ultimate through
a UK proxy.
Use the UK proxy: Link:
http://www.docoja.com/blue
Insert this link into the proxy: Link:
http://www.hotukdeals.com/freebies/b...e-box-p/834553
Then click on the burnout paradise picture on the right that says "Go to
deal". And that should bring you to the checkout in EA store!
Borrowed from CAG.
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w*******y
发帖数: 60932 | 48 This is ripped directly from CAG and I thought I'd share and do my part to
increase the foot traffic into Kmart. Deals may be live on Kmart.com but I
would NOT count on anything working unless it is reflected on their website
but that'll most likely only be for games that are reduced in price, not for
promotional offers.
Here is the list:
Deals valid 01/30 - 02/05
PS3 160GB $50 Kmart Gift Card
PSP 3000 $129.99 (Save $40)
Two Worlds II $39.99 (Reg. $59.99)
God of War 3 and God of War Collection $... 阅读全帖 |
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w*******y
发帖数: 60932 | 49 From Cag: Link:
http://www.cheapassgamer.com/forums/showthread.php?t=286158
quote
Hello All, Here again with next weeks ad.
All Kinect Games 2 For $70
including: EA Sports Active 2 - $50
Buy One Video Game, Get The Second For 50% Off 19.99 or less.
Music Bluray
Rush: Beyond The Lighted Stage - 16.99
Metallica: The Big Four - Live From Sofia, Bulgaria - 16.99
U2: 360At The Rose Bowl - 16.99
Eric Clapton: Crossroads Guitar Festival 2010 - 16.99
The Rolling Stones: Ladies & Gentlemen - 14.99
Michae... 阅读全帖 |
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w*******y
发帖数: 60932 | 50 (Stolen from CAG:
http://www.cheapassgamer.com/forums/showthread.php?p=8126856
)
PS3 Sale
Droplitz Sale (PS3) (now $4.99, original price $9.99)
PSP Sale
Class of Heroes Legacy Sale (PSP) (now $19.99, original price $39.99)
Crimson Gem Saga Sale (PSP) (now $14.99, original price $29.99)
Hammerin Hero Sale (PSP) (now $9.99, original price $19.99)
Hexyz Force Sale (PSP) (now $14.99, original price $29.99)
Kenka Bancho: Badass Rumble Sale (PSP) (now $19.99, original price $39.99)
Knights in th... 阅读全帖 |
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