审稿人让补充实验，but客观条件不允许(附模板)

　　花了很多时间和金钱做出来的实验数据，投稿之后，审稿人说要让补充实验，虽然很难以接受，但是又不能眼看着到手的接收函飞了，如果审稿人要求了补充实验，最好还是去做这个实验，然后补充一些数据。

　　如果客观条件就是没有办法及时补充实验，直接回复审稿人不能补充，那八成会被拒稿。这种情况下也不要捉急，在返修时可以真诚的告诉审稿人，当时没有做这个实验的原因，同时现在因为条件限制做不了实验，或者向审稿人说明白自己认为不需要做这个实验的原因都是可以的。

　　回复模板都给大家准备好啦

　　我常用的回复格式:

　　Dear reviewer:

　　I am very grateful to your comments for the manuscript. According with

　　your advice, we amended the relevant part in manuscript. Some of your

　　questions were answered below. 1)

　　2)

　　....

　　引用审稿人推荐的文献的确是很重要的，要想办法和自己的文章有机地结合起

　　来。

　　至于实验大部分都可以不用补做，关键是你要让审稿人明白你的文章的重点是什

　　么，这个实验对你要强调的重点内容不是很必要，或者你现在所用的方法已经可

　　以达到目的就行了。

　　最后要注意，审稿人也会犯错误，不仅仅是笔误也有专业知识上的错误，因为编

　　辑找的审稿人未必是你这个领域的专家。只要自己是正确的就要坚持。在回复中

　　委婉地表达一下你的意见，不过要注意商讨语气哦!

　　我得回复格式是这样的：

　　Dear Professor xx:

　　Thank you very much for your letter dated xxx xx xxxx, and the referees’

　　reports. Based on your comment and request, we have made extensive

　　modification on the original manuscript. Here, we attached revised

　　manuscript in the formats of both PDF and MS word, for your approval. A

　　document answering every question from the referees was also summarized

　　and enclosed.

　　A revised manuscript with the correction sections red marked was attached

　　as the supplemental material and for easy check/editing purpose.

　　Should you have any questions, please contact us without hesitate.

　　然后再附上 Q/A，基本上嘱条回答，写的越多越好(老师语)。结果修改一次就

　　接收了：)

　　我的回复，请老外帮忙修改了

　　Dear Editor:

　　Thank you for your kind letter of “......” on November **, 2005. We

　　revised the manuscript in accordance with the reviewers’ comments, and

　　carefully proof-read the manuscript to minimize typographical,

　　grammatical, and bibliographical errors.

　　Here below is our description on revision according to the reviewers’

　　comments.

　　Part A (Reviewer 1) 1. The reviewer’s comment: ......

　　The authors’ Answer: .....

　　2. The reviewer’s comment: ......

　　The authors’ Answer: .....

　　...

　　...

　　Part B (Reviewer 2)

　　1. The reviewer’s comment: ......

　　The authors’ Answer: .....

　　2. The reviewer’s comment: ......

　　The authors’ Answer: .....

　　...

　　...

　　Many grammatical or typographical errors have been revised.

　　All the lines and pages indicated above are in the revised manuscript.

　　Thank you and all the reviewers for the kind advice.

　　Sincerely yours,

　　***

　　一个回复的例子(已接收)

　　Major comments:

　　1. The authors need to strengthen their results by including MMP

　　secretion, and tran-matrigel migration by a positive control progenitor

　　cell population i.e. enriched human CD34 cells obtained from mobilized

　　PBL, since this is a more clinically relevant source of CD34 cells which

　　has also been shown to secrete both MMP-9 and MMP-2 (ref. 11). CD34

　　enriched cells from steady state peripheral blood which also secrete MMPs

　　are also of interest.

　　2. In fig 1C please specify which cell line represents

　　MMP-negative cells. This needs to be clarified, as well as a better

　　explanation of the method of the protocol.

　　3. The ELISA results are represented as "fold increase" compared

　　to control. Instead, we suggest that standards should be used and results

　　should be presented as absolute concentrations and only then can these

　　results be compared to those of the zymography.

　　4. When discussing the results, the authors should distinguish

　　clearly between spontaneous migration vs chemotactic migration.

　　Furthermore, the high spontaneous migration obtained with cord blood CD34 cells should be compared to mobilized PBL CD34 enriched cells and

　　discussed.

　　5. The authors claim that the clonogenic assay was performed to

　　determine the optimum concentration for inhibition of MMP activity by

　　phenanthroline and anti MMP-9 mAb, however they should clarify that this

　　assay can only determine the toxicity of the inhibitors and not their

　　optimal inhibitory concentrations.

　　Minor comments:

　　1. There are many spelling and syntax errors, especially in the

　　results and discussion, which need correction.

　　a. Of special importance, is the percent inhibition of migration,

　　which is described as percent of migration. i.e. pg 7:"Migration of CB

　　CD34 was reduced to 73.3%?" Instead should read "Migration of CB CD34 was

　　reduced by 73.3%?"

　　b. The degree symbol needs to be added to the numbers in Materials

　　and methods.

　　2. It would be preferable to combine figure 1A and B, in order

　　to confirm the reliability of fig. 1B by a positive control (HT1080).

　　Answer to referee 1 comment:

　　1. Mobilized peripheral blood is a more clinical source of CD34+

　　cells, so it is necessary to compare the MMP-9 secretion and

　　trans-migration ability of CB CD34+ cells with that of mobilized PB CD34+

　　cells. However, we couldn't obtain enough mobilized PB to separate PB

　　CD34+ cells and determine the MMP-9 secretion and migration ability, so

　　we couldn’t complement the study on PB CD34+ cells in this paper. Results

　　obtained by Janowska-Wieczorek et al found that mobilized CD34+ cells in

　　peripheral blood express MMP-9. Furthermore, Domenech’s study showed that

　　MMP-9 secretion is involved in G-CSF induced HPC mobilization. Their

　　conclusions have been added in the discussion. In our present study, our

　　central conclusion from our data is that freshly isolated CD34+

　　stem/progenitor cells obtained from CB produce MMP-9.

　　2. MMP-9 negative cell used in fig 1C was Jurkat cell. In

　　zymographic analysis, MMP-9 was not detected in the medium conditioned

　　by Jurkat cell. To exclude that the contaminating cells may play a role

　　in the observed MMP-9 production, we screened the media conditioned by

　　different proportion of CB mononuclear cells with MMP-9 negative cells

　　by zymography. This result may be confusion. Actually, only by detecting

　　the medium conditioned by 2X105 CB mononuclear cells (MNC)/ml (since the

　　purities of CD34+ cell are more than 90%), it could exclude the MNC role. In the revised manuscript, we only detected MMP-9 activity and antigen

　　level in the medium conditioned by 2X105 CB mononuclear cells (MNC)/ml.

　　There is no MMP-9 secretion be detected in the medium conditioned by 2X105

　　CB MNC/ml. It excluded the possibility that the MMP-9 activity in CB CD34+

　　cells conditioned medium is due to the contamination by MNC.

　　3.In this revised paper, we have detected the MMP-9 antigen

　　levels by using commercial specific ELISA kits (R&D System, sensitivity,

　　0.156ng/ml). Recombinant MMP-9 from R&D System was used as a standard.

　　The results are expressed in the absolute concentration. The absolute

　　concentration result has been added in the paper. As shown in Fig2, MMP-9

　　levels were detectable in both CB CD34+ cell conditioned medium and BM

　　CD34+ cell conditioned medium. However, MMP-9 level was significantly

　　higher in CB CD34+ cell conditioned medium than in BM CD34+ cell

　　conditioned medium (0.406±0.133ng/ml versus 0.195±0.023ng/ml).

　　Although gelatinolytic activity was not detected in media conditioned by

　　CD34+ cells from BM, sensitivity of ELISA favors the detection of MMP-9

　　antigen in the BM CD34+.

　　4. In our study, to establish the direct link between MMP-9 and

　　CB CD34+ cells migration, we only determined the role of MMP-9 in

　　spontaneous migration of CB CD34+ cells, but not in chemotactic migration.

　　Actually, regulation of hematopoietic stem cell migration, homing and

　　anchorage of repopulation cells to the bone marrow involves a complex

　　interplay between adhesion molecules, chemokines, cytokines and

　　proteolytic enzymes. Results obtained by the groups of Voermans reveal

　　that not only the spontaneous migration but also the SDF-1 induced

　　migration of CB CD34+ cells is greatly increased in comparison to CD34+

　　cells from BM and peripheral blood.

　　5. CD34+ cells we obtained in each cord blood sample were very

　　limited. It is not enough to screen the inhibitors concentrations to

　　select the optimal inhibitory concentrations. In the blocking experiments,

　　based on the concentrations used by others and the manufacturer's

　　recommendation, we then determined the inhibitors concentrations by

　　excluding the toxicity of the inhibitors in that concentration, which was

　　determined by clonogenic assay.

　　Minor comments:

　　1.The spelling and syntax errors have been checked and corrected.

　　2.Since the results in figure 1A and B were obtained from two separated

　　and parallel experiments, it is not fitness to combine two figures.

　　回复审稿人的时候还要注意2个原则：

　　1、就算审稿人意见有点没道理，也要礼貌的回复，态度尽量不要强硬;

　　2、即便真的有些审稿意见不合理，可以提出自己的想法，找一些证据来证明自己的合理性。

　　回复审稿人一定要真诚、有礼貌，审稿人的每个意见都要一一回复，实在没办法补充实验，就真诚的说出自己的难处，也要表示对审稿意见的肯定，然后表明自己今后会继续研究的决心，不过如果是高分期刊，这一招可能不太好使，让补充还是要乖乖补充实验的~

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