g*****n
发帖数: 250 | 1 I speculated, yesterday, how they picked NTCP as a candidate receptor for
HBV. There is nothing wrong to select a candidate based on knowledge. The
key is to determine whether that is the one you are looking for with
evidence. Let’s see how they tested whether NTCP is the receptor for HBV.
(1) They transfected NTCP into cells, cross-linked the viral proteins,
and ran Western. Again, overexpression, forced linking, all are too
artificial. They should look at the native NTCP protein, but they never did.
Funny thing is: this is an almost a repeat of the first experiment. How did
they think that this could add something meaningful on top of the first
experiment?
(2) They transfected NTCP-GFP into cells, cross-linked red-fluorescence-
labeled c viral proteins, and observed their co-localization in the cells.
Who, on earth, thinks that this is way to show protein-protein interaction??
!! You transfect the cells green and force-link red to the cells. They
surely are present in the same cells. This is no brainer. Does that mean
that they interact with zero distance? Give me a break.
Also, they seemed to have no clue what this method does. They show images of
low magnification. Why? This is not a quantitative assay. How many cells
are lit up does not matter. You should look at the cells as close as
possible to see subcellular localization of the staining. Even you use high
power objectives, the regular fluorescent microscopy is the wrong way to go.
You need a confocal microscope to scan the cell to make sure that the
signals are in the membrane. Even you see them in the membrane, it does not
indicate that the proteins interact. The bottom line is that this is a wrong
method to answer the question. How come those PhDs did not understand the
methodology!!
When I look at those pictures, I feel like an audience watching a lousy
magic show. You know it's fake and you know the tricks, but they go on
performing anyway.
(3) They knocked down NTCP in cells using siRNA and assessed viral
infection and propagation. They show that the level of NTCP mRMA is
decreased by siRNA. Who cares about the mRNA? The level of NTCP protein is
what matters in this case. There is no guarantee that lower mRNA level
results in lower protein level. However, they never show what happened to
the protein of NTCP. This just takes a couple of runs of Western blotting.
Why not did it?
(4) Even NTCP siRNA affected HBV infection and propagation, as shown in
the paper, it does not mean that NTCP is the receptor. It just simply means
that the siRNA changes cellular conditions, which, in turn, influence viral
propagation. It does even not suggest that NTCP is directly associated with
the viruses.
Do I have to complete the rest of the paper? I rather go to watch Youtube to
kill time.
If this a showcase of China’s science from the showcase Institute, I feel
sorry for the taxpayers. | w***r
发帖数: 709 | 2 这都什么乱七八糟的，兄弟咱改行干别的行不
did.
did
【在 g*****n 的大作中提到】
: I speculated, yesterday, how they picked NTCP as a candidate receptor for
: HBV. There is nothing wrong to select a candidate based on knowledge. The
: key is to determine whether that is the one you are looking for with
: evidence. Let’s see how they tested whether NTCP is the receptor for HBV.
: (1) They transfected NTCP into cells, cross-linked the viral proteins,
: and ran Western. Again, overexpression, forced linking, all are too
: artificial. They should look at the native NTCP protein, but they never did.
: Funny thing is: this is an almost a repeat of the first experiment. How did
: they think that this could add something meaningful on top of the first
: experiment?
| n********y
发帖数: 187 | 3 我觉得LZ的再读是非常丰富详细系统的技术细节讨论---对很多读者,内行和外行都是科
普读物.另外maggieklean 的有关病毒传染机制的讨论也从大的机制方向上给了很好的
科普.
LZ和maggieklean (还有本WSN)的用词很尖刻,但是绝对不是上来就否定WNEHUI的一切,
而是从具体科学上讨论.
这里是中国人自己的BBS,我不认为这样的讨论是诽谤和恶毒攻击.
铁牛的IFs则是从参与WENHUI文章的研究人员的TRACK RECORD上来反驳具体详细的质疑-
--好象我们是在做人身攻击.
我对RY的评论还算是人身攻击,但是对于这WENHUI文章科学的讨论仅仅是学习. | g*****n
发帖数: 250 | 4 没有证明NTCP是HVB的受体，那基本上就全盘否定了。 |
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