Change of mean concentration of inflammatory cytokines in plasma from baseline to day 60, day 90, and day 120.

<p>A) IL-1β; B) IL-6; C) IL-17; D) TNF-α. Data are presented in the CMS+fluoxetine group (n = 9), CMS+vehicle group (n = 9) and control group (n = 6). Wilcoxon signed-rank test was performed to investigate the time effect. *<i>p</i><0.05 <i>vs</i>. baseline.</p

Change of mean body weight from baseline to day 120.

<p>Data are presented in the CMS+fluoxetine (0.042mg/g once daily dissolved in 0.5ml distilled water per rat) group (n = 9), the CMS+vehicle (0.5ml distilled water per rat once daily) group (n = 9) and the control group (n = 6) respectively. Paired-sample t-test was used for the statistical analysis. ***<i>p</i><0.001, **<i>p</i><0.01 <i>vs</i>. baseline. CMS = chronic mild stress.</p

MiR-339-5p Regulates the Growth, Colony Formation and Metastasis of Colorectal Cancer Cells by Targeting PRL-1

<div><p>MicroRNAs (miRNAs) have been suggested to play a vital role in regulate tumor progression and invasion. However, the expression of miR-339-5p in colorectal cancer and its effects are not known. Here, we report that miR-339-5p is a tumor suppressor by regulating expression of PRL-1. In this study, we showed that downregulated miR-339-5p levels in colorectal cancer tissues and highly invasive CRC cell lines. Furthermore, enhancing the expression of miR-339-5p inhibited CRC cell growth, migration and invasion in vitro and suppressed tumor growth in vivo. We then screened and identified a novel miR-339-5p target, phosphatases of regenerating liver-1 1 (PRL-1), and it was further confirmed by luciferase assay. Overexpression of miR-339-5p would also reduce the expression of PRL-1 mRNA and protein. The reduced PRL-1 expression was associated with low expression of phosphorylated-extracellular signal-regulatedkinase1/2 (p-ERK1/2). Conversely, reduction of miR-339-5p by inhibitors in cells stimulated these phenotypes. In conclusion, our results demonstrate that miR-339-5p functions as a tumor suppressor and plays a role in inhibiting growth and metastasis of CRC cells through targeting PRL-1 and regulating p-ERK1/2 .These findings suggest that miR-339-5p may be useful as a new potential therapeutic target for CRC.</p></div

Comparison of time and group difference of behavioral test scores.

<p>A) Mean percentage of sucrose preference at baseline and on day120 in the CMS+fluoxetine group (n = 9), CMS+vehicle group (n = 9) and control group (n = 6) by paired-sample t-test. *<i>p</i><0.05 <i>vs</i>. baseline. B) The effect of FST for CMS+fluoxetine group, CMS+vehicle group and control in the mean immobility time recorded on day 120 by one-way ANOVA. Bars represent a mean ± standard error. CMS = chronic mild stress; FST = force swim test; ANOVA = analysis of variance.</p

Comparison of mean levels of inflammatory cytokines in the brain on day 120.

<p>Comparison of mean levels of inflammatory cytokines in the brain on day 120.</p

Comparison of mean plasma levels of inflammatory cytokines from day 0 to day 120.

<p>Comparison of mean plasma levels of inflammatory cytokines from day 0 to day 120.</p

The effects of administration of fluoxetine and vehicle on IL-1β levels (day 120) in the brains of rats subjected to an animal model of chronic mild stress.

<p>Bars represent a mean+standard error. Control rats were not subjected to any stress or treatment. *<i>p</i><0.05 <i>vs</i>. CMS+vehicle group according to Kruskal-Wallis test followed by Mann-Whitney U test.</p

HMGB3 promote CRC cells proliferation and migration in vitro.

<p>(A) qRT-PCR analysis of HMGB3 expression in six CRC cell lines. The differences between independent experimental groups were tested by using Independent-Samples t-test. Error bars indicate mean ± SD of 3 independent experiments. ***, <i>p</i> < 0.001. (B) qRT-PCR and western blot analyses of HMGB3 mRNA and protein levels after the transfection of the sh-HMGB3 or HMGB3 plasmid. The differences between independent experimental groups were tested by using Independent-Samples t-test. ***, <i>p</i> < 0.001. (C) Effects of sh-HMGB3 and oe-HMGB3 on cell proliferation were determined by CCK8 cell proliferation assay. Error bars represent the mean ± SD of 5 independent experiments. The differences between independent experimental groups were tested by using Independent-Samples t-test. ***, <i>p</i> < 0.001. (D) The proliferation ability was determined by colony formation assay of the cell SW620 and HT29 after inhibiting the expression of HMGB3. The bar chart represents the colony number. Error bars represent the mean ± SD of 3 independent experiments. The differences between independent experimental groups were tested by using Independent-Samples t-test. **, <i>p</i> < 0.01; ***, <i>p</i> < 0.001. (E) Cell cycle G1 arrest after knocking down HMGB3 measured by flow cytometry. (F) The migration capacity was determind by transwell assays. The bar chart represents the migration cell numbers. Error bars represent the mean ± SD of 5 different field. The differences between independent experimental groups were tested by using Independent-Samples t-test. ***, <i>p</i> < 0.001. (G) Effects of sh-HMGB3 or oe-HMGB3 on cell migration ability were determined by Wound healing assay.</p

Expression levels of HMGB3 mRNA and protein are increased in CRC tissues.

<p>(A) qRT-PCR analysis of HMGB3 expression in 34 paired CRC tissues and adjacent normal tissues. HMGB3 expression level was normalized to GAPDH and the results were presented as the fold-change in tumor tissues relative to the matched adjacent normal tissues. Error bars indicate mean ± standard deviation of 3 independent experiments. (B) The value of ΔCt was used to show the expression level of HMGB3 (ΔCt = Ct_HMGB3 –Ct_GAPDH) in the 34 paired human CRC tissues (T) and adjacent normal tissues (N) (<i>p</i> < 0.05). The differences between tumor group and normal group were tested by using a Paired-samples <i>t</i>-test. (C) Western blot analysis of HMGB3 protein expression in seven pair CRC tissues compared with the normal tissues. The differences between tumor group and normal group were tested by using a Independent-Samples t-test. (D) Representative HMGB3 staining in CRC specimens. Left, normal tissue, Scale bars <i>=</i> 50 μm. Middle, CRC tissues, Scale bars = 50μm. Right, CRC tissues, Scale bars <i>=</i> 20 μm. The HMGB3 index was calculated as that the number of HMGB3 positive cells divided by the number of total cells ×100% (magnification, ×200). The differences between tumor group and normal group were tested by using a Independent-Samples t-test. Error bars represent the mean ± SD of 5 different fields. ***, <i>p</i> < 0.001.</p

Table_1_Lower insulin level is associated with sarcopenia in community-dwelling frail and non-frail older adults.pdf

BackgroundSarcopenia is common among older individuals with and without type 2 diabetes mellitus (T2DM). There are conflicting evidence in support of the role of insulin in the development of age-related and T2DM-related sarcopenia. We investigated the relationships between the levels of fasting insulin and other blood biomarkers related to insulin or lipid metabolism with the presence of sarcopenia in two independent studies.Materials and methodsIn 246 pre-frail frail older individuals with (n = 41) and without T2DM (n = 205) in the Singapore Frailty Interventional Trial, sarcopenia was defined by low appendicular lean mass (ALM) relative to total body mass (skeletal muscle index, SMI = ALM/height2) and low lower limb strength or gait speed according to the Asian Working Group for Sarcopenia (AWGS) criteria released in 2019, and related to levels of fasting insulin and glucose, C-peptide, IGF-1, leptin, and active ghrelin. This investigation was validated in another independent study sample of 189 robust and pre-frail frail elderly in the Singapore Longitudinal Aging Study Wave 2 (SLAS-2).ResultsCompared to non-sarcopenic individuals, those with sarcopenia and possible sarcopenia showed significantly lower fasting insulin (p ConclusionDysregulated insulin secretion in diabetic and non-diabetic older individuals may play an important role in age-related and diabetes-related sarcopenia.</p