Assessments of contamination and human health risks of heavy metals in the road dust from a mining county in Guangxi, China

<p>A total of 32 road dust samples were collected from mining areas and a control area in Nandan County, Guangxi, in order to investigate the contamination characteristics of heavy metals and associated health risks to local residents. The results indicated that elevated concentrations of As, Cd, Cu, Pb, Sb, and Zn were found in road dust in mining areas compared with control areas and background values. Pearson's correlation analysis and principal component analysis indicated that As, Cd, Cu, Pb, Sb, and Zn in road dust mainly originated from anthropogenic sources (e.g., vehicles emission, mining activities, and smelting activities), whereas Cr, Co, and Ni were associated with natural sources (e.g., soil weathering). Furthermore, noncarcinogenic hazards to both adults and children were found in mining areas, while noncarcinogenic health risks in the control area were negligible. The health hazard was mostly ascribed to the ingestion, followed by the dermal contact, and the inhalation. The cancer risks from As, Cd, Co, Cr, and Ni in all studied areas were within safe levels as the <i>R</i> values were below the threshold of 1 × 10⁻⁶.</p

Effect of AMPK siRNA on the expression of differentiation transcriptional factors and lipogenic protein.

<p>Post-confluent 3T3-L1 cells were differentiated and treated with 10 µM UA for 6 days after the silencing of AMPK. PPAR_γ, C/EBP_α and FAS, pACC and ACC expressions were assessed by Western blotting as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070135#s2" target="_blank">Materials and Methods</a>. Data are expressed as means ± SD (n = 3). *P<0.05 and **P<0.001 vs. the corresponding controls.</p

Effect of UA on the protein expression of lipogenic proteins,

<p>Cpt1, AMPK and Sirt1. 3T3-L1 preadipocytes were incubated in differentiation medium without or with different concentrations of UA (added on day 0 of differentiation) for 6 days. The expression of pACC, ACC, FAS, FABP4, Cpt1, pAMPK, AMPK and Sirt1 were assessed by Western blotting as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070135#s2" target="_blank">Materials and Methods</a>. Data are expressed as means ± SD (n = 3). *P<0.05 and **P<0.001 vs. the corresponding controls.</p

Effect of UA on lipid accumulation in 3T3-L1 adipocytes.

<p>(A) Post-confluent 3T3-L1 preadipocytes were induced to differentiate in the absence or presence of UA (added on day 0 of differentiation) for 6 days. The morphological changes associated with cell differentiation were photographed after Oil Red O staining. (B) Stained lipids were extracted and quantified by measuring absorbance at 520 nm. Data are expressed as means ± SD (n = 3). * P<0.05 and ** P<0.001 vs. the control.</p

Effects of UA on the viability, proliferation, cell cycle and apotosis of 3T3-L1 preadipocytes.

<p>(A) 3T3-L1 preadipocytes were incubated in differentiation medium with or without UA. After 6 days, MTT reagent was added to the medium. After 4 hours of incubation, the medium was aspirated and 150 µL DMSO was added to each well. The absorbance was read at 570 nm. (B) 3T3-L1 preadipocytes were seeded onto 96-well plates at a density of 3×10³ cell/cm² and treated with indicated concentrations of UA for 24, 48 and 72 hours, respectively. MTT assay was performed as described above to reflect the proliferation. (C) 3T3-L1 preadipocytes were seeded onto 24-well plates at a density of 3×10³ cell/cm² and treated with different concentrations of UA for 24, 48 and 72 hours, respectively. The number of adherent cells was determined using an automatic cell counter. (D, E) Three days post-confluence, 3T3-L1 preadipocytes were differentiated in the presence or absence of 2.5, 5, 10 µM ursolic acid for 24 hours. The cells were collected and fixed overnight with 70% ethanol at 4°C, and stained with propidium iodide solution. Cell cycle and apoptosis rate were analyzed on a flow cytometry. Data are expressed as means ± SD (n = 3). * P<0.05 and ** P<0.001 vs. the control.</p

Effect of UA on the protein expression of differentiation-related transcriptional factors.

<p>(A)–(B) 3T3-L1 preadipocytes were incubated in differentiation medium without or with different concentrations of UA (added on day 0 of differentiation) for 6 days. The expression of PPAR_γ and C/EBP_α were assessed by Western blotting as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070135#s2" target="_blank">Materials and Methods</a>. (C) The C/EBP_β expression was determined after preadipocytes were incubated in differentiation medium in the presence or absence of different concentrations of UA for 2 days. (D) The SREBP-1c expression was determined after preadipocytes were incubated in differentiation medium in the presence or absence of 10 µM UA for 6 days. Data are expressed as means ± SD (n = 3). *P<0.05 and **P<0.001 vs. the corresponding controls.</p

Effect of AMPK siRNA or Sirt1 inhibitor on lipid accumulation in 3T3-L1 adipocytes.

<p>(A–C) Post-confluent 3T3-L1 cells were induced for differentiation and treated with 10 µM UA (added on day 0 of differentiation) in the absence or presence of AMPK siRNA or 10 mM nicotinamide for 6 days. Images were captured using an inverted microscope after Oil Red O staining. (D) Stained lipids were extracted and quantified by measuring absorbance at 520 nm. Data are expressed as means ± SD (n = 3). *P<0.05 and **P<0.001 vs. the corresponding controls; ^#P<0.05 and ^## P<0.001 vs. nicotinamide treated cells.</p

Additional file 4 of Genome-wide prediction and functional analysis of WOX genes in blueberry

Supplementary Material

Additional file 3 of Genome-wide prediction and functional analysis of WOX genes in blueberry

Supplementary Material

Additional file 5 of Genome-wide prediction and functional analysis of WOX genes in blueberry

Supplementary Material