h*******o
发帖数: 4884 | 1 For whole brain or hemisphere, what I did is to overnight fix them at 4oC.
Change to PBS with sodium azide for storage afterwards.
Make sure that your brain samples were perfused well.
Residue serum/blood is usually the cause of high background.
Also, block for longer time may help too.
Last take a 3D stack then get sharper images through deconvolution. |
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b****e
发帖数: 52 | 4 因为样品宝贵,取了一半脑做WB,一半固定(没有灌流),结果发现anti-mouse 2nd Ab识别
很多血管的非特异信号.请问有什么办法可以降低background吗?只能选择灌流吗?出生
几天(P10)的小鼠也能灌流吗? 谢谢了 |
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S******9
发帖数: 2837 | 5 脑子如果灌注以后很容易没有背景的
10天的老鼠可以，直接左心室灌注，用30G的针头
现在可以试一下blocking buffer： 5%BSA+5%FBS+0.3%Triton 100 |
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g*****n
发帖数: 250 | 7 在一抗前，用anti-m 抗体封蔽。脑子都抠出来了，切一半做WB, 那还怎么灌？ |
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b****e
发帖数: 52 | 8 谢谢大家.我现在用的是5%NGS,0.1%Tx100 blocking, 有什么区别吗? HRP coupled
anti-mouse Ab 可以用来做封蔽吗?我们lab没有uncoupled Ab. 1:5000 1hr? |
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h*******o
发帖数: 4884 | 9 换个rabbit的抗体是最简单的方法
或者就像楼上说的，用anti－mouse IgG block |
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g*****n
发帖数: 250 | 10 在5%NGS,0.1%Tx100溶液里加点抗体。NGS还得要。HRP coupled
anti-mouse Ab 可以用，只要你不用其底物来检测。1:500－1000 1hr。 |
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l********o
发帖数: 134 | 12 不知道大家知不知道这家公司的这个系统。求各位大牛能够说说，教教我这个系统治疗
癌症靠铺嘛？
Chemosaturation of the liver represents an evolution from a highly invasive
open surgical procedure to a minimally invasive procedure for treating
patients with cancers in the liver.
In hepatic chemosaturation therapy, a percutaneous hepatic perfusion (PHP)
procedure delivers ultra-high doses of intra-arterial chemotherapy directly
into the isolated liver, saturating both the liver and the tumor cells. The
blood from the liver is drained through an i... 阅读全帖 |
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w****o
发帖数: 781 | 13 多谢！请问大家每次杀老鼠取tissue之前都做perfusion么？ |
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s*****i
发帖数: 315 | 14 这个完全取决于你的实验目的啊，如果你发现血液干扰，当然要perfuse，如果无所谓
，当然怎么方便怎么来 |
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h*******o
发帖数: 4884 | 15 做不做perfusion, 要看实验设计，有些必须做，有些不能做 |
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l*******n
发帖数: 554 | 16 我只能帮你大致说下。
一台显微镜是需要的，从普通的到共聚焦，价格不一而且差几十倍，我想国内都查得到。
Micromanipulator $15000
Patch clamp amplifier $20000
Software $2000
Perfusion system $2500
Image chamber $500
一下想不起来polish的仪器要多少钱，应该不是很贵，就算$1000
耗材(假设你要屯很多） $5000
还有细胞培养的一些费用，人工费，杂七杂八的费用得靠你自己算了。
我不知道在国内买的话是否会加价很多，这个你也要考虑到。 |
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p*******r
发帖数: 4048 | 17 不会贵很多，而且你还可以买国产的，便宜大概一半。仪博产的Amplifier其实还不错
。周专的老师开的公司。防震平台的话连胜就可以了，不用进口。
manipulator国产没有好的，进口吧。perfusion国产的够了。
如果你需要具体报价私下联系我，但不能透露你哪弄来的价，因为我当年都花了不少功
夫砍价的。 ：） |
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y******2
发帖数: 326 | 18 之所以存在都是有原因的，要不然真是玩儿的？美国5，6十年代拿猫狗杀着玩儿的你是
没看到。我在迈阿密确实以为IACUC都是在捣乱的，那是因为大家都遵纪守法，IACUC纯
增加负担。但是到了纽约某学校才知道IACUC是有必要的，某AAALAC执照被吊销的学校
动物房那不是玩儿的，我这么邋遢的人都看不下去了，连perfusion室都要我自己现建
，好几百年的学校了。都tmd影响到实验人员的健康问题了。 |
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y******2
发帖数: 326 | 19 之所以存在都是有原因的，要不然真是玩儿的？美国5，6十年代拿猫狗杀着玩儿的你是
没看到。我在迈阿密确实以为IACUC都是在捣乱的，那是因为大家都遵纪守法，IACUC纯
增加负担。但是到了纽约某学校才知道IACUC是有必要的，某AAALAC执照被吊销的学校
动物房那不是玩儿的，我这么邋遢的人都看不下去了，连perfusion室都要我自己现建
，好几百年的学校了。都tmd影响到实验人员的健康问题了。 |
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l*******n
发帖数: 554 | 20 楼主是说抽取brochoalveolar lavage fluid吗？
如果是的话，我们是在气管处减一小口，不能剪断，然后把一钝口针头伸入小口中，用
棉线将气管和针头扎紧，防止漏，接着将PBS注射入小鼠肺中，不用太多，看肺快要完
全扩展开来后停止，然后慢慢将BALF抽回针筒。
我们是用它来测细胞因子，病毒模型没做过，看看文献吧，看看有没其他人做过。 |
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n*********s
发帖数: 552 | 21 谢谢答复，不过不是BAL，就是貌似注射器灌好hbss，然后从心脏打进去，hbss从心脏
进入肺里，肺就鼓起来了。是要去掉red blood cell吗？那为什么不用RBC lysis
buffer呢？ |
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T****u
发帖数: 424 | 22 necroptosis和influenza感染，好topic~
KO mice有difference吗？Nec-1能protect吗？ |
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n*********s
发帖数: 552 | 23 真是牛哥啊，看出俺是做这个的了。目前只做过一次pilot expt，还真有点差异，nec
－1还没试。想看下lung里的immune response，打算rt－pcr测下lung里cytokines，就
不知道harvest lung的时候有什么讲究没有？这年月我做的简直是天马行空，好多东西
都是现学啊～ |
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T****u
发帖数: 424 | 25 你得把里面的血都去掉，否则一堆macrophage inflitration，测出来都是高的。
拿个免疫细胞的基因一起跑做参照。
有个问题RIP1/3在里面表达吗？
nec |
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A******y
发帖数: 2041 | 26 Hypoxia is lack of oxygen, And oxidative stress is about dysfunction of our
OXPHOS pathway. Hypoxia injury is still from oxdative damage during the re-
perfusion phase. |
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s****9
发帖数: 932 | 27 B cell normally do not infiltrate lung after flu infection. B cell localize
in the draining LNs to secrete antibody. LN stromal cells have high level
of CXCL13 and BAFF to support B cell homeostasis. Unlike T cells, B cells
also do not need to reach tissue to exert the function. The secreted
antibody goes through blood and lung is highly vascularized.
Have you perfuse the lung to move leukocytes from blood? B cells are quite
sensitive to apoptosis and growth factors. I would first check ... 阅读全帖 |
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e**o
发帖数: 345 | 28 谢谢。你说的这些信息很有用，都是我不知道的
收集细胞之前，做了perfuse
怎么检测全身的B cell percentage
再次感谢。
localize
level
quite |
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r*******e
发帖数: 123 | 29 Biotechniques. 2003 Sep;35(3):472-4, 476.
Modifications of a commercial perfusion system for use in ultrafast solution
exchange during patch clamp recording.
Hinkle DJ1, Bianchi MT, Macdonald RL.
PMID: 14513550
万分感谢。请e-mail: [email protected]
/* */ |
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k*****e
发帖数: 372 | 31 hypocortisolism 导致 hypoNa 主要因为Ald不够，不能保Na
hypothyroidism导致hypoNa的原因不完全清楚，但还是主要是SIADH，有个理论是甲低
导致CardiacOutput低，renal perfusion不足，ADH就过度分泌，导致低钠
这是reference：
Hanna, FW, Scanlon, MF. Hyponatraemia, hypothyroidism, and role of arginine-
vasopressin. Lancet 1997; 350:755 |
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s*****9
发帖数: 445 | 32 it says"one month later", also it already tells you that both the FEV1 &
stress TEST ARE NORMAL, so the pulmonary function should be OK.
Besides, This patient w/ primary pulmonary hypertension undergoes lung transplant to improve the perfusion, which has nothing to do with her pulmonary function itself.
I 'm with Answer B.
procedure,
new
to |
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u***e
发帖数: 611 | 33 为啥降压，书上不是说不超220不用降吗，会减少perfusion影响康复。 |
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W******g
发帖数: 143 | 34 Summary
In summary, cardioversion is performed on perfusing arrhythmias. The goal is
not to cause VF. Therefore, during cardioversion, the shock is administered
at the peak of the R-wave, during the absolute refractory period. Delivery
of the shock during the relative refractory period can cause the development
of a nonperfusing arrhythmia such as VF.
Cardioversion is performed to treat unstable SVT due to re-entry, unstable
atrial fibrillation, and unstable atrial flutter. These arrhythmias are |
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k**********n
发帖数: 438 | 35 老柳细节讲得很好了，俺胡了八图昏了一年了，早听受益更多。
我个人体会是要抓住主要矛盾以及看住不良倾向。主治大夫基本也就关注主要矛盾，你
把主要矛盾cover了，他们对其他小节就纠结的少一些。例如，嗯嗯，例如...
感染你得说血象，band, left shift, trend.你还得说fever, 细菌培养。你还得说病
人从哪里来的，从家里来的那就是community aquired，从nursing home来的那就是
hospital aquired,抗生素的用法就不一样了。说sepsis你得说血压的趋势，脉搏(80那
可能不是septic shock，130你得考虑把心律降下来了，ventricular filling要不灵光
了)。尿量(ml/hr)，反映peripheral perfusion，line，CVP,液体出入量. Bicarb,gap
, lactic acid，还有用药。
CHF那你得说病因，diastolic还是systolic，EF多少。体重，尿量，lasix用量，肺湿
不湿，腿肿不肿，O2 sat行不行，dig level， K, Mg.
COPD你对说吸 |
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u***e
发帖数: 611 | 36 关于PE和V/Q scan我上次说V/Q很不准现在发现也不对，有必要更新一下，免得又误人
子弟，毁人不倦了。。。。。
前几天收了一个病人，syncope进来的，ER做了V/Q, intermediate probability，我一
拿到又傻眼了，这个病人没有SOB,chest pain, 做V/Q只是因为发现有点desaturation
，而Cr是1.8. 临床上属于那种low probability的那种。想忽略V/Q吧，又有点怕。只
好去找了radiologist，到底你什么意思，给个这么个报告。后来放射科医生给我们上
了一课，觉得很有收获，什么PIOPED study，什么样的算作是intermediate
probability，并不是他们拿不准。好的内科医生还是得会看片子，结合病人的胸片，
以前的CT，才能对V/Q的结果作出正确的判断。那个病人有明显的perfusion defect但
是还是没有达到high probability的标准，也没有肺部本身的病变。还是高度怀疑PE的
。于是我们就上了抗凝，然后超声就发现了腿部的DVT。
最近的NEJM上有一篇关于PE的综述，有 |
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S******9
发帖数: 2837 | 37 nbme4 b3-34
the graph shows coronary blood flow as a function of perfusion pressure.
the solid line indicates the relationship between blood flow and
pressure when the coronary arteries are maximally dilated.which of the
following pairs of lettered points best illustrates the change that
occurs when a healthy person at rest begins to run to catch a bus?
1. w-->Y
2.X--->Y
3.X--->Z
4.Y--->W
5.Z--->Y
从rest到run，HR increased, contranction increased----> SV increased--->CO
increased---> coronary flow |
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z****o
发帖数: 368 | 38 Thanks Springstep. you are the expert! Another Q in follow up. Is the
coronary perfusion pressure equal to the aortic diastolic pressure? |
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l********y
发帖数: 2295 | 39 B1Q14. An 80-year-old man has had poor balance for 6 months. He has a
history of hypertension treated with hydrochlorothiazide. His blood pressure
is 136/86 mm Hg. Neurologic examination shows mild tremor of the hands when
his arms are outstretched and decreased vibratory sensation at the knees.
Deep tendon reflexes of the quadriceps and gastrocnemius-soleus muscles are
hyperactive. Babinski's sign is present bilaterally. He is unable to stand
with his eyes closed. Which of the following is most... 阅读全帖 |
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n***h
发帖数: 364 | 40 Answer: A
Diagnosis: secondary HTN.
Clues: Stenosis of right renal artery--- low perfusion in kidney---renin
level increased---RAA system activated---HTN.
Key: RAA level is diff. in primary and secondary HTN. |
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l*******8
发帖数: 290 | 41 Sorry, there's a figure in the question stem. The question is about coronary
blood flow and perfusion pressure. What's the change when a person at rest
begins to run to catch a bus?
Any input will be appreciated. |
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w******n
发帖数: 61 | 42 Fluid balance is studied in a blood-perfused muscle preparation with an
intact lymphatic system from an experimental animal. After a control period
during which all
parameters are normal, increasing which of the following is most likely to
increase the rate of lymph flow without affecting the protein content of the
lymph?
A) Capillary colloid osmotic pressure
B) Capillary hydrostatic pressure
C) Constriction of precapillary sphincters
D) Endothelial transcytosis
E) Width of junctions between end... 阅读全帖 |
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L********r
发帖数: 37 | 43 45. A 32-year-old woman comes to the physician because of a 4-month history
of fatigue, cough, and shortness of breath with exertion. She has had two
episodes of pneumonia and one episode of severe sinusitis over the
past 2 years. She has never smoked. She takes no medications. Crackles are
heard at the left lung base. An x-ray film of the chest shows a left lower
lobe infiltrate and scarring of the right base.
A) Measurement of serum a1-antitrypsin level
B) Methacholine challenge test
C) Quanti... 阅读全帖 |
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d****y
发帖数: 2180 | 44 选A还是E？
Paracentesis 虽说是比较general 的抽液，在这题可以理解为心包抽液吗？
18. A 50-year-old woman has had progressive dyspnea over the past 2
weeks
and constant, sharp chest pain for 4 days. The pain is localized to the
center of the chest and is worse while supine. She underwent a right,
modified radical mastectomy and adjuvant chemotherapy for breast cancer
3
years ago. She has a history of hypothyroidism treated with thyroid
replacement therapy. She has smoked one pack of cigarettes daily for 30
years and drinks t... 阅读全帖 |
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J**b
发帖数: 182 | 45 some more:
7.“reciproca” change in lead I with inferior wall STeMI:
A. Elevated ST segment
B. Low-voltage QRS
C. Wide QRS
D. Depressed ST segment
E. Irregular narrow QRS
8. Ventricular ectopy:
A. Elevated ST segment
B. Low-voltage QRS
C. Wide QRS
D. Depressed ST segment
E. Irregular narrow QRS
9. All of the following are true EXCEPT
A. The AV node is usually perfused by the posterior descending coronary
artery
B. The right ventricular bundle has an ante... 阅读全帖 |
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t******a
发帖数: 408 | 46 Intraosseous infusion (IO)
病人 has MCA stroke, 同时还有junctional arrhythmia, HR42, low urine output,
dehydration.
在tibia上打一个孔，接上infusion fluids, and meds.
works well for adult patients.
顿时想到UW（CK？）有一题考的就是IO。只不过是儿童病人。 现在adult patients 也
用。 让我感觉到UW出的题目还是比较紧扣临床实践，考查实际中重要的知识。
我的感受UW不光考试有用，对临床也很有帮助。
*************************
下面摘自medscape
http://emedicine.medscape.com/article/908610-overview
IO techniques have fewer serious complications than central lines and can be
performed much faster than central or ... 阅读全帖 |
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a********n
发帖数: 438 | 49 有一点小小疑问啊，这个病人不是came in with severe stroke with hx of CHF,
ESRD,
HTN, DM and so on. 怎么后来又表示自己血压低？就这个病人的情况，她是STROKE进
来的，需要保持血压高点，才能保持brain perfusion. 还有，她是ESRD/CHF, 也是要
很小心的给fluid bolus.最好是医生能够review一下她的药，找出她血压持续低的原因。
vital
pressure |
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s*******y
发帖数: 1739 | 50 你可以不把RISK的东西放第一DIAGNOSIS,但Ineffective tissue perfusion 肯定是有
了,肯定比你那个FATIGUE是PRIORITY,我感觉你对NURSING DIAGNOSIS的POINT还是没抓
住,不是病人现在稳定,她的PRIORITY就放在那些她现在现有的问题上,Preeclampsia还
是很凶险的,头十分钟稳定,后十分钟可能胎儿就宫内窘迫了,胎儿的缺氧也不是单纯
late deceleration能看到的,PREECLAMPSIA对胎盘造成的影响是逐步的,不断的.你休息
一下,再好好想想是不是,GOOD LUCK. |
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