Reduced ATP production by Mfn2 knockdown or inhibition of autophagic degradation.

<p>Intracellular ATP production of HeLa cells (<b>A</b>) infected with scrambled RNA, Mfn2 shRNA, or Mfn2 shRNA co-infected with Mfn2 cDNA, (<b>B</b>) transfected with scrambled RNA, Rab7 siRNA1, or Rab7 siRNA2, or (<b>C</b>) treated with autophagic degradation inhibitors Bafilomycin A1, 3-Methyladenine (3-MA), or NH4Cl. n = 4 independent experiments for each group. *, p<0.05 versus control cells; #, p<0.05 versus Mfn2 shRNA group.</p

Ferroelectric Mesocrystalline BaTiO₃/SrTiO₃ Nanocomposites with Enhanced Dielectric and Piezoelectric Responses

A platelike mesocrystalline BaTiO₃/SrTiO₃ (BT/ST) nanocomposite is prepared via a clever two-step solvothermal soft chemical process. First a protonated titanate H_1.07Ti_1.73O₄·<i>n</i>H₂O (HT) crystal with a layered structure and platelike morphology is solvothermally treated in a Ba­(OH)₂ solution to generate a homogeneous platelike BaTiO₃/HT (BT/HT) nanocomposite. Second the generated BT/HT nanocomposite is solvothermally treated in a Sr­(OH)₂ solution to generate the mesocrystalline BT/ST nanocomposite with platelike particle morphology. The transformation reactions from the HT precursor to the mesocrystalline BT/ST nanocomposite are topochemical conversion reactions, and the formed BT/ST nanocomposite is constructed from well-aligned BT and ST nanocrystals in the same crystal-axis orientation. The BT/ST nanocomposite annealed at 900 °C shows a ferroelectric behavior and drastically enhanced piezoelectric and dielectric responses owing to the introduction of a lattice strain at a three-dimensional heteroepitaxial interface between the BT and ST nanocrystals in the mesocrystal. The nanostructure of the BT/ST mesocrystal is suitable for simultaneous application of the strain engineering and the orientation engineering to develop high performance piezoelectric and dielectric materials

Mfn2 knockdown impairs autophagic degradation.

<p>(<b>A</b>) Increased LC3-II level in Mfn2 knockdown HeLa cells by Western blot. n = 4 independent experiments for each group. (<b>B</b>) LC3-II:GAPDH ratios in HeLa cells infected with scrambled RNA, Mfn2 shRNA, or Mfn2 shRNA with Mfn2 cDNA. n = 3 independent experiments for each group. *, p<0.05 versus scramble control; ^#, p<0.05 versus Mfn2 shRNA group. (<b>C</b>) Western blot showing LC3-II levels in scramble and Mfn2 shRNA cells with or without starvation treatment in the presence or absence of lysosomal inhibitor Bafilomycin A1 (BA). (<b>D</b>) Confocal images of HeLa cells transfected with GFP-mRFP-LC3 plasmid, and co-infected with adenovirus containing scrambled RNA or Mfn2 shRNA, with or without starvation for 36 hours (left), and percentages of merged green and red LC3 dots to total LC3 dots. Scale bar: 10 μm. n = 20 cells for each group from 3 independent experiments. *, p<0.05 & **, p<0.01 versus scramble control; ^#, p<0.05 versus scramble starvation group. (<b>E</b>) mRNA level of Tom1 and Lamp2a by real-time PCR in HeLa cells transfected with Pcmv-Tom1 or Pcmv-Lamp2a plasmids. n = 3 independent experiments. (<b>F</b>) Fold changes of cell counting by CCK8 in HeLa cells infected with adenovirus containing scrambled RNA co-transfected with Pcmv vector or Pcmv-Tom1, Pcmv-Lamp2a, and Mfn2 shRNA, Mfn2 shRNA co-transfected with Pcmv-Tom1 or Pcmv-Lamp2a. n = 3 independent experiments. *, p<0.05 versus control; ^#, p<0.05 versus Mfn2 shRNA only group.</p

Mfn2 knockdown inhibits cell proliferation.

<p>(<b>A</b>) Western blotting showing Mfn2 and Mfn1 protein levels in HeLa cells infected with adenovirus containing scrambled RNA or two sets of Mfn2 shRNAs. β-actin was used as protein loading control. (<b>B</b>) Confocal imagings of HeLa cells stained with mitoTrackor showing mitochondrial morphology. Scale bar: 10 μm. (<b>C</b>) Cell counting kit-8 (CCK8) assay of HeLa cells without infection or infected with adenovirus containing scrambled RNA or Mfn2 shRNA. (<b>D</b>) CCK8 assay of T/G HA-VSMC cells infected with adenovirus containing scrambled RNA or Mfn2 shRNA at time point as indicated after infection. (<b>E</b>) 3-(4,5–dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay of HeLa cells infected with scrambled RNA, Mfn2 shRNA, or Mfn2 shRNA co-infected with Mfn2 cDNA. (<b>F</b>) Cell apoptosis assay by flow cytometry in HeLa cells infected with scrambled RNA or Mfn2 shRNA. (<b>G</b>) Fluorescence activated cell sorting analysis by flow cytometry to measure cell cycle distribution in HeLa cells infected with scrambled RNA, Mfn2 shRNA, or Mfn2 shRNA co-infected with Mfn2 cDNA, and (<b>H</b>) the histogram plot. n = 3–6 independent experiments for each group. *, p<0.05 versus scramble control.</p

Inhibited glycolysis by Mfn2 knockdown.

<p>(<b>A</b>) Traces of extracellular acidification rates (ECAR) of HeLa cells in response to mitochondrial inhibitors. (<b>B</b>) Average data of basal ECAR and that in the presence of mitochondrial inhibitors as in <b>A</b>. (<b>C</b>) ECAR of HeLa in response to glucose, oligomycin, and 2-deoxy-D-glucose (2-DG). (<b>D</b>) ECAR showing glucose metabolism and glycolytic capacity. n = 3 independent experiments for each group. *, p<0.05 versus scramble control.</p

BMI z-score in relation to urinary phthalate metabolite concentrations, by median (ppb) quartiles for ∑All in 8-10yrs boys (A), MBP and ∑LMP in 11-13yrs boys (B), and ∑MEHP in 8-10yrs girls (C).

<p>Obesity and normal weight children were stratified by age- and gender-specific weight distribution based on the national survey in Chinese school children, normal weight children (<80th %) and obesity children (≥90th%), National Puberty Timing and Health Effects in Chinese Children (PTHEC) study, Shanghai 2010-2012.</p

Associations between exposure quartiles for log-transformed concentrations of MBP and ∑LMP and body fat distribution indices with significant continuous regression coefficients in 11-13yrs boys of China, corrected for puberty onset, socio-economic level, physical activity and dietary nutriment intake.

<p>(Blue rhombus: regression coefficient; Error bar: 95% confidence interval; log-transformed indices including subscapular skinfold thickness, triceps skinfold thickness and BF ratio; BMI, BMI z-score, waist circumference, hip circumference, and BSA were normally distributed data)</p

Associations between exposure quartiles for log-transformed concentrations of MEHHP and ∑All monoesters and body fat distribution indices with significant continuous regression coefficients in 8-10yrs boys of China, corrected for puberty onset, socio-economic level, physical activity and dietary nutriment intake.

<p>(Blue rhombus: regression coefficient; Error bar: 95% confidence interval; log-transformed indices including subscapular skinfold thickness, triceps skinfold thickness and BF ratio; BMI, BMI z-score, waist circumference, hip circumference, and BSA were normally distributed data)</p

Associations between log-transformed urinary phthalate metabolite concentrations and overweight/obesity in 493 school children of China.

a<p>OB means obesity, OV means overweight, NO means normal weight.</p>b<p>Logistic regression analysis adjusted for socio-economic level, physical activity, dietary nutriment intake and puberty onset, phthalate metabolite concentrations as continuous variables, p<0.05.</p>c<p>Logistic regression analysis of variance adjusted for socio-economic level, physical activity, dietary nutriment intake and puberty onset, phthalate metabolite concentrations as ordinal variables, p<0.05.</p

Age and sex-specific phthalate metabolite concentrations (µg/L) in urine samples of school children.

a<p>Detection frequency.</p>b<p>Analysis of variance adjusted for sex, p<0.05.</p>c<p>Analysis of variance adjusted for age, p<0.05.</p