The effect of EPO on the levels of insulin signaling events in the liver.

<p>The levels of total Akt, IR and IRS1 and phosphorylated Akt, IR and IRS1 in the livers of individual mice were determined by Western blot assays and the relative levels of Akt, IR and IRS1 phosphorylation were analyzed by densimetric analysis using ImageJ software. Data shown are representative images and expressed as the mean ± SEM of individual groups (n = 6 per group) of mice from three separate experiments. (<b>A</b>) Western blot and quantitative analyses of the levels of Akt activation. (<b>B</b>) Western blot and quantitative analyses of the levels of IR activation. (<b>C</b>) Western blot and quantitative analyses of the levels of IRS1 activation. *P<0.05 vs. the HFD-Con group.</p

The EPOR mRNA transcription in the livers of mice.

<p>Total RNA was extracted from the liver and kidney samples of individual mice and reversely transcribed into cDNA. The EPOR mRNA transcripts were detected by semi-quantitative RT-PCR. Data shown are representative images of agarose gel electrophoresis from three separate experiments.</p

The effect of EPO on the expression of PEPCK and G6Pase in the liver.

<p>The relative levels of PEPCK and G6Pase in the livers of individual mice were determined by quantitative RT-PCR and Western blot assays. Data are representative images or expressed as the mean ± SEM of individual group (n = 8 per group) of mice from three separate experiments. (<b>A</b>) RT-PCR analysis of the relative levels of PEPCK and G6Pase mRNA transcripts to control GAPDH in the liver. (B) Western blot and quantitative analysis of the relative levels of PEPCK in the liver. (<b>C</b>) Western blot and quantitative analysis of the relative levels of G6Pase in the liver. ^#P<0.05 vs. the NC group, *P<0.05 vs. the HFD-Con group.</p

The effect of EPO on the expression of inflammatory signaling events in the liver.

<p>The levels of activated NF-<i>κ</i>B p65 in the liver were determined by DNA binding-based ELISA (<b>A</b>). The relative levels of TLR4 expression were characterized by RT-PCR (<b>B</b>) and Western blot assays (<b>C</b>). The relative levels of JNK (<b>D</b>), ERK (<b>E</b>) and p38 MAPK (<b>F</b>) phosphorylation in the liver were characterized by Western blot assay and densimetric analysis. Data are representative images or expressed as the mean ± SEM of each group (n = 6) of mice from three separate experiments. ^#P<0.05 vs. the NC group, *P<0.05 vs. the HFD-Con group.</p

The effect of EPO on HFD-fed mice.

<p>C57BL/6 mice were fed with HFD for 12 weeks and treated with EPO (HFD-EPO) or injected with saline (HFD-Con) for two weeks. A control group (NC) of mice was fed with normal chow and injected with saline. The body weights and amounts of food consumed in individual mice were measured at the indicated time points. At the end of treatment, the mice were fasted and subjected to measurements of the levels of fasting blood glucose, fasting serum insulin and IPGTT. The AUC for blood glucose in individual mice was calculated. Data are present as the mean ± SEM of each group (n = 8) of mice from two-three separate experiments. (<b>A</b>) The body weights. (<b>B</b>) The levels of fasting blood glucose. (<b>C</b>) The levels of fasting serum insulin. (<b>D</b>) The glucose tolerance. (<b>E</b>) The values of AUC. There was no significant difference in the amounts of food intake among these groups of mice (data not shown). ^#P<0.05 or ^##P<0.01 vs. the NC group, *P<0.05 or **P<0.01 vs. the HFD-Con group.</p

The sequences of the primers.

<p>The sequences of the primers.</p

The effect of EPO on inflammatory cytokine expression in mice.

<p>At the end of treatment, blood samples were obtained from individual mice and the levels of serum TNF-α and IL-6 were measured by ELISA. In addition, the relative levels of TNF-α and IL-6 mRNA transcripts to control GAPDH were analyzed by quantitative RT-PCR. Data are expressed as the mean ± SEM of the concentrations of serum cytokines or the relative levels of cytokine mRNA transcripts to control GAPDH in each group (n = 8) from three separate experiments. (A) ELISA for the levels of serum TNF-α. (B) ELISA for the levels of serum IL-6. (C) RT-PCR analysis of the relative levels of cytokine mRNA transcripts. ^#P<0.05 vs. the NC group, *P<0.05 vs. the HFD-Con group.</p

DataSheet_1_Placenta-derived exosomes exacerbate beta cell dysfunction in gestational diabetes mellitus through delivery of miR-320b.docx

Recent studies have shown placenta-derived exosome (pdE) acts as an important mediator of organ-to-organ interplay regulating maternal metabolic alterations, however, the function and mechanisms of placental exosomes on pancreatic β-cell maladaptation in gestational diabetes mellitus (GDM) remain unclear. The purpose of this investigation was to ascertain how placental exosomes affected the β-cell dysfunction associated with the onset of GDM. Exosomes were isolated from chorionic villi explants of pregnant mice and humans with normal glucose tolerance (NGT) and GDM. The effects of pdE from GDM on glucose tolerance in vivo and islets function in vitro were determined. Isolated islets from mice fed on the chow diet displayed an increase in apoptosis and observed their glucose-stimulated insulin secretion (GSIS) greatly diminished by PdE from GDM mice. Mice that accepted PdE from mice with GDM possessed glucose intolerance.Based on miRNA microarray assay and bioinformatics analysis from human placental exosomes, we identified miR-320b selectively enriched in PdE secreted in GDM compared with NGT. Importantly, the level of placental miR-320b was positively correlated with the 1h-glucose and 2-h glucose of a 75 g oral glucose tolerance test (OGTT) during human pregnancies. Furthermore, miR-320 overexpression attributed to impaired insulin secretion and increased apoptosis in MIN6 cells and islets obtained from mice with normal insulin sensitivity. This study firstly proposed that altered miRNAs in pdE contribute to defective adaptation of β cells during pregnancy, which expands the knowledge of GDM pathogenesis. Exosomes from the placenta may be an emerging therapeutic target for GDM.</p

Global epidemiology of diabetic foot ulceration: a systematic review and meta-analysis^†

<p>Diabetic foot is a severe public health issue, yet rare studies investigated its global epidemiology. Here we performed a systematic review and meta-analysis through searching PubMed, EMBASE, ISI Web of science, and Cochrane database. We found that that global diabetic foot ulcer prevalence was 6.3% (95%CI: 5.4–7.3%), which was higher in males (4.5%, 95%CI: 3.7–5.2%) than in females (3.5%, 95%CI: 2.8–4.2%), and higher in type 2 diabetic patients (6.4%, 95%CI: 4.6–8.1%) than in type 1 diabetics (5.5%, 95%CI: 3.2–7.7%). North America had the highest prevalence (13.0%, 95%CI: 10.0–15.9%), Oceania had the lowest (3.0%, 95% CI: 0.9–5.0%), and the prevalence in Asia, Europe, and Africa were 5.5% (95%CI: 4.6–6.4%), 5.1% (95%CI: 4.1–6.0%), and 7.2% (95%CI: 5.1–9.3%), respectively. Australia has the lowest (1.5%, 95%CI: 0.7–2.4%) and Belgium has the highest prevalence (16.6%, 95%CI: 10.7–22.4%), followed by Canada (14.8%, 95%CI: 9.4–20.1%) and USA (13.0%, 95%CI: 8.3–17.7%). The patients with diabetic foot ulcer were older, had a lower body mass index, longer diabetic duration, and had more hypertension, diabetic retinopathy, and smoking history than patients without diabetic foot ulceration. Our results provide suggestions for policy makers in deciding preventing strategy of diabetic foot ulceration in the future.Key messages</p><p>Global prevalence of diabetic foot is 6.3% (95%CI: 5.4–7.3%), and the prevalence in North America, Asia, Europe, Africa and Oceania was 13.0% (95%CI: 10.0–15.9%), 5.5% (95%CI: 4.6–6.4%), 5.1% (95%CI: 4.1–6.0%), 7.2% (95%CI: 5.1–9.3%), and 3.0% (95% CI: 0.9–5.0%).</p><p>Diabetic foot was more prevalent in males than in females, and more prevalent in type 2 diabetic foot patients than in type 1 diabetic foot patients.</p><p>The patients with diabetic foot were older, had a lower body mass index, longer diabetic duration, and had more hypertension, diabetic retinopathy, and smoking history than patients without diabetic foot.</p><p></p> <p>Global prevalence of diabetic foot is 6.3% (95%CI: 5.4–7.3%), and the prevalence in North America, Asia, Europe, Africa and Oceania was 13.0% (95%CI: 10.0–15.9%), 5.5% (95%CI: 4.6–6.4%), 5.1% (95%CI: 4.1–6.0%), 7.2% (95%CI: 5.1–9.3%), and 3.0% (95% CI: 0.9–5.0%).</p> <p>Diabetic foot was more prevalent in males than in females, and more prevalent in type 2 diabetic foot patients than in type 1 diabetic foot patients.</p> <p>The patients with diabetic foot were older, had a lower body mass index, longer diabetic duration, and had more hypertension, diabetic retinopathy, and smoking history than patients without diabetic foot.</p

Clinical and biochemical characteristics of 18350 patients with Type 2 diabetes and with diagnosed hypertension.

<p>*, Median (25^th percentile to 75^th percentile) and their P values were derived from Two Sample Wilcoxon test.</p