Title: Expression deregulation of matrix metalloproteinases and vasoconstriction related genes
in Pakistani females with abnormal uterine bleeding
Version: 2 Date: 04 Oct 2022
Reviewer's report:
The manuscript (78d90311-9d7d-4afa-bb5f-b1cfb4d34e66) entitled “Expression analysis of
different genes in Pakistani females with abnormal uterine bleeding” by Akram et al. aimed
to investigate the expression profile of targeted genes to explore the pathogenesis of abnormal
uterine bleeding (AUB) in blood samples collected from women with AUB or aged matched
control.
AUB describes irregularities in the menstrual cycle altering in the frequency, regularity,
duration, and volume. Up to one-third of women will experience abnormal uterine bleeding in
their life, with irregularities most commonly occurring at menarche and perimenopause. The
researches explain the molecular and cellular causes of AUB pathogenesis are important to
evaluate and treat abnormal uterine bleeding.
The manuscript is poorly written, needs to improve and revised many sentences by correcting
many grammatical errors. The sentences are confusing, should be clearly presented. Please see
my comments in below:
1. The authors should clarify: a) why they do choice qPCR instead of ELISA that measures serum
levels of these targets in collected blood samples; b) why they did not get endometrial biopsies
to measure the expression levels of targeted gene; and c) why these five genes are selected??
2. The authors should revise the Figure 1 and 2 because control group is represented by the bars
whereas AUB group is represented by line.
3. Also, Figure 1A is totally unclear, it should be revised.
4. In Figures 1 and 2, the authors should be explained how they get lower standard variations or
errors in both control and AUB groups in a working a larger group /N=112.
5. The authors found increased MMP9 and VEGF mRNA levels in married women with AUB
versus single women with AUB. The authors should explain how marriage status affects the
expression of these genes. Are these married women using any contraceptives? if yes, what kind
of contraceptives such as pills, IUD, etc., they are using??
6. In Figure 2C, the authors compared the expression levels of target genes in AUB group
with miscarriage history to AUB without miscarriage history, found that expression of MMP2
and TGF# were significantly increased, whereas levels of MMP9 and VEGF were significantly
decreased in AUB females with miscarriage history compared to AUB females without
miscarriage history. The authors explain why the number in the bars are lower. If there is a
significant difference in these group, standard derivation or errors should be higher in Figure 1
that displays a cumulative group.
7. Page 9, Line 2, the sentence should be edited, it would be “compared to AUB females without
miscarriage history”
8. The mean ±SD or SEM values should be given in the Results section.
9. PTGFR is a receptor for PGF2a, the regulation of this gene is important for endometrial and
myometrial as well as ovarian function. The author should explain why they include this gene
their list in blood samples.
10. What are the basal endogenous levels of targeted genes in peripheral blood mononuclear
cells (PBMCs)?
11. The sequences of primers, product size of each target gene should be provided.
12. The author clearly explain how they analyzed qPCR data.
13. All abbreviations in the manuscript should be used in first cited sentence and be consistent
through manuscript.
14. All gene names should be revised based on HUGO nomenclators.
15. Discussion was very poor written. The authors should discuss their results, clearly citing the
importance of their findings by comparing the previous studies.
16. The number in each group should be added in each figure.