Reviewer's report
Title:HMGB1 cytoplasmic translocation in patients with acute liver failure
Version: 1Date:16 December 2010
Reviewer: Hiroki Kuniyasu
Reviewer's report:
Major Compulsory Revisions
1. In Figure 1, viability ratio was indicated to assess the cell damage by LPS and
TNF-alpha. HepG2 cells were growing cells. The cell number should be indicated
in addition to the viability ratio.
2. In Figure 1D and F, extracellular HMGB1 was induced with the optimal dosage
of LPS and TNF-alpha. How the authors translate the phenomenon?
3. In Figure 2C, extracellular HMGB1 should be added to cytoplasmic an nuclear
localization of HMGB1 to assess the HMGB1 distribution.
4. In Figure 2C, the cytoplasmic HMGB1 was increased; however, the nuclear
HMGB1 kept the level. The cytoplasmic HMGB1 is translocated from nucleus or
newly translated? At least, mRNA expression of HMGB1 should be examined.
5. For induce liver damage in HepG2 cells, LPS and TNF-alpha were used.
HMGB1 is well known as an inducer of secretion of TNF-alpha, INF-gamma, and
IL-1beta, which form a vicious cycle. Then the extracellular concentrations of
TNF-alpha, INF-gamma, and IL-1beta should be examined to be compared
between treatments with LPS and TNF-alpha.
6. In Figure 4, the disease status of chronic hepatitis B is completely different
from that of acute liver failure by HBV. The details of clinical data showing how
chronic or how acute should be needed for this type of comparison. The present
form of the data is afraid to be less values.
7. In Figure 4, the histology of acute liver failure showed marked necrotic
reactions. Many hepatocytes showed unintegrated cell structure, which is not
suitable for judging the HMGB1 location. Fig. 4 should be omitted from the
manuscript.
8. The cytoplasmic localization of HMGB1 was clearly indicated by
immunostaining (Fig. 5B); however, the cytoplasmic HMGB1-positive
hepatocytes were scatted in the lobes. The characterization of these cells is
needed.
9. In Fig. 5f, some cytoplasmic HMGB1-positive hepatocytes also showed the
nuclear HMGB1 but some cells did not. What is the different between the two
types of HMGB1 localization?
10. The expression of IL-8 and GRO should be examined in the D-GalN-induced
mouse hepatitis to compare with HMGB1 localization.
Level of interest: An article of importance in its field
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests.
Reviewer's report
Title:HMGB1 cytoplasmic translocation in patients with acute liver failure
Version: 1Date:16 December 2010
Reviewer: Hiroki Kuniyasu
Reviewer's report:
Major Compulsory Revisions
1. In Figure 1, viability ratio was indicated to assess the cell damage by LPS and
TNF-alpha. HepG2 cells were growing cells. The cell number should be indicated
in addition to the viability ratio.
2. In Figure 1D and F, extracellular HMGB1 was induced with the optimal dosage
of LPS and TNF-alpha. How the authors translate the phenomenon?
3. In Figure 2C, extracellular HMGB1 should be added to cytoplasmic an nuclear
localization of HMGB1 to assess the HMGB1 distribution.
4. In Figure 2C, the cytoplasmic HMGB1 was increased; however, the nuclear
HMGB1 kept the level. The cytoplasmic HMGB1 is translocated from nucleus or
newly translated? At least, mRNA expression of HMGB1 should be examined.
5. For induce liver damage in HepG2 cells, LPS and TNF-alpha were used.
HMGB1 is well known as an inducer of secretion of TNF-alpha, INF-gamma, and
IL-1beta, which form a vicious cycle. Then the extracellular concentrations of
TNF-alpha, INF-gamma, and IL-1beta should be examined to be compared
between treatments with LPS and TNF-alpha.
6. In Figure 4, the disease status of chronic hepatitis B is completely different
from that of acute liver failure by HBV. The details of clinical data showing how
chronic or how acute should be needed for this type of comparison. The present
form of the data is afraid to be less values.
7. In Figure 4, the histology of acute liver failure showed marked necrotic
reactions. Many hepatocytes showed unintegrated cell structure, which is not
suitable for judging the HMGB1 location. Fig. 4 should be omitted from the
manuscript.
8. The cytoplasmic localization of HMGB1 was clearly indicated by
immunostaining (Fig. 5B); however, the cytoplasmic HMGB1-positive
hepatocytes were scatted in the lobes. The characterization of these cells is
needed.
9. In Fig. 5f, some cytoplasmic HMGB1-positive hepatocytes also showed the
nuclear HMGB1 but some cells did not. What is the different between the two
types of HMGB1 localization?
10. The expression of IL-8 and GRO should be examined in the D-GalN-induced
mouse hepatitis to compare with HMGB1 localization.
Level of interest: An article of importance in its field
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests.
Please review this paper.
user
Reviewer's report
Title:HMGB1 cytoplasmic translocation in patients with acute liver failure
Version: 1Date:16 December 2010
Reviewer: Hiroki Kuniyasu
Reviewer's report:
Major Compulsory Revisions
1. In Figure 1, viability ratio was indicated to assess the cell damage by LPS and
TNF-alpha. HepG2 cells were growing cells. The cell number should be indicated
in addition to the viability ratio.
2. In Figure 1D and F, extracellular HMGB1 was induced with the optimal dosage
of LPS and TNF-alpha. How the authors translate the phenomenon?
3. In Figure 2C, extracellular HMGB1 should be added to cytoplasmic an nuclear
localization of HMGB1 to assess the HMGB1 distribution.
4. In Figure 2C, the cytoplasmic HMGB1 was increased; however, the nuclear
HMGB1 kept the level. The cytoplasmic HMGB1 is translocated from nucleus or
newly translated? At least, mRNA expression of HMGB1 should be examined.
5. For induce liver damage in HepG2 cells, LPS and TNF-alpha were used.
HMGB1 is well known as an inducer of secretion of TNF-alpha, INF-gamma, and
IL-1beta, which form a vicious cycle. Then the extracellular concentrations of
TNF-alpha, INF-gamma, and IL-1beta should be examined to be compared
between treatments with LPS and TNF-alpha.
6. In Figure 4, the disease status of chronic hepatitis B is completely different
from that of acute liver failure by HBV. The details of clinical data showing how
chronic or how acute should be needed for this type of comparison. The present
form of the data is afraid to be less values.
7. In Figure 4, the histology of acute liver failure showed marked necrotic
reactions. Many hepatocytes showed unintegrated cell structure, which is not
suitable for judging the HMGB1 location. Fig. 4 should be omitted from the
manuscript.
8. The cytoplasmic localization of HMGB1 was clearly indicated by
immunostaining (Fig. 5B); however, the cytoplasmic HMGB1-positive
hepatocytes were scatted in the lobes. The characterization of these cells is
needed.
9. In Fig. 5f, some cytoplasmic HMGB1-positive hepatocytes also showed the
nuclear HMGB1 but some cells did not. What is the different between the two
types of HMGB1 localization?
10. The expression of IL-8 and GRO should be examined in the D-GalN-induced
mouse hepatitis to compare with HMGB1 localization.
Level of interest: An article of importance in its field
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests.
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