EVI-1 is involved in miR-206/CALR in human breast cancer stem cells.

(A&B) breast cancer stem cells were transfected with Ad-EVI-1 or si-EVI-1 for 48 h, the EVI-1 expression was determined by immunoblot analysis; (C&D) Human breast stem cell were treated with miR-206 mimics, miR-206 mimics+Ad-EVI-1 and Ad-EVI-1 for 48 h. EVI-1 and CALR expression were measured by Western blotting analysis; (E), the mRNA level of EVI-1 and CALR were measured by real-time quantitative PCR; (F), Schematic of MiR/206-EVI-1/CALR pathway in human breast cancer. All results are expressed as the mean ± S.D. from three independent experiments, *P .05, **P .01; #P.05, ##P.01 vs. control group.</p

Overexpression of CALR attenuate the cytotoxic effect of miR-206 on growth of breast cancer stem cells.

Human breast cancer stem cells were treated by miR-206 and/or Ad-CALR for 48 h, then the cells were harvested and analyzed. (A&B) CALR protein expression were determined by immunoblot analysis; (C) Human breast cancer stem cells were transfected with miR-206 and/or Ad-CALR for 72 h, the cell growth was determined by MTT assay; (D&E) Effect of CALR overexpression attenuate the apoptosis effect of miR-206 in breast cancer stem cells. Human breast cancer stem cells were treated by miR-206 and/or Ad-CALR for 48 h, the early and late apoptotic cells were measured by PI-Annexin V, *P < 0.05, **P < 0.01 vs. control group.</p

Effect of miR-206 on CALR expression in breast cancer stem cells.

Human breast cancer stem cells were treated with various concentrations of miR-206 mimics (0, 30, 50, 70 nM) for 48 h. CALR protein expression (A&B) and mRNA expression (C) level were measured by Western blot and real-time quantitative PCR, respectively; Human breast cancer stem cells were treated with 50 nM miR-206 mimics for 0, 24, 48 and 72h. CALR protein expression (D&E) and mRNA expression level were determined by Western blot and real-time quantitative PCR, respectively. All results are expressed as the mean ± S.D. from three independent experiments. *P .05, **P .01 vs. control group.</p

Raw images for western blots.

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Effect of miR-206 on CALR-mediated metastasis of human breast cancer stem cells.

Cells were treated with miR-206 mimics, miR-206 mimics+Ad-CALR and Ad-CALR for 48 h, respectively. (A) invasive breast cancer stem cells were counted; (B&D), CALR protein expression was measured by Western blotting analysis; C, CALR gene was measured by real-time quantitative PCR. All results are expressed as the mean ± S.D. from three independent experiments, *P .05, **P .01; #P.05, ##P.01 vs. control group.</p

Sub-10 nm Hexagonal Lanthanide-Doped NaLuF₄ Upconversion Nanocrystals for Sensitive Bioimaging in Vivo

By thermal decomposition in the presence only of oleylamine, sub-10 nm hexagonal NaLuF₄-based nanocrystals codoped with Gd³⁺, Yb³⁺, and Er³⁺ (or Tm³⁺) have been successfully synthesized. Sub-10 nm β-NaLuF₄: 24 mol % Gd³⁺, 20 mol % Yb³⁺, 1 mol % Tm³⁺ nanocrystals display bright upconversion luminescence (UCL) with a quantum yield of 0.47 ± 0.06% under continuous-wave excitation at 980 nm. Furthermore, through the use of β-NaLuF₄:Gd³⁺,Yb³⁺,Tm³⁺ nanocrystals as a luminescent label, the detection limit of <50 nanocrystal-labeled cells was achieved for whole-body photoluminescent imaging of a small animal (mouse), and high-contrast UCL imaging of a whole-body black mouse with a penetration depth of ∼2 cm was achieved

Characterization of circulating CD14⁺CD169⁺ monocytes in CRC patients.

<p>Peripheral blood mononuclear cells (PBMCs) were obtained from individual subjects and stained with anti-CD14 and anti-CD169. The frequency of circulating CD14⁺CD169⁺ monocytes was characterized by flow cytometry. The cells were gated initially on mononuclear cells and then on CD14⁺ cells. Subsequently, the percentages of CD14⁺CD169⁺ monocytes were determined. To characterize different subsets of CD14⁺CD169⁺ monocytes, PBMCs were stained with fluorescent antibodies against CD14, CD169 and CD163, CD206 or MAC387. The cells were gated on CD14⁺CD169⁺ monocytes and the percentages of CD14⁺CD169⁺CD163⁺, CD14⁺CD169⁺CD206⁺ and CD14⁺CD169⁺MAC387⁺ macrophages were determined. Data are representative charts of flow cytometry and expressed as the values of individual subjects. The horizontal lines indicate the median for individual groups. (A) Flow cytometry analysis of CD14⁺CD169⁺ monocytes/macrophages. (B) Flow cytometry analysis of different subsets of circulating CD14⁺CD169⁺ monocytes/macrophages. (C) The percentages of CD14⁺CD169⁺ monocytes. (D) The frequency of CD14⁺CD169⁺CD163⁺ macrophages. (E) The frequency of CD14⁺CD169⁺CD206⁺ macrophages. (F) The frequency of CD14⁺CD169⁺MAC387⁺ macrophages.</p

Stratification analysis of the percentages of CD14⁺CD169⁺ circulating monocytes and tumor infiltrating macrophages in CRC patients.

<p>The CRC patients were stratified as early (I/II, n = 21) or advanced stage (III/IV, n = 25) and the percentages of CD14⁺CD169⁺ circulating monocytes and TIMs in individual patients were analyzed. Furthermore, the potential association between the percentages of CD14⁺CD169⁺ circulating monocytes or TIMs and the levels of plasma CEA in individual groups of patients were analyzed. Data are the mean of individual subjects. (A) Percentages of CD14⁺CD169⁺ monocytes in PBMCs (n = 21 for early stage, n = 25 for advanced stage of CRC patients) and the percentages of CD14⁺CD169⁺ macrophages in TIMs from early (n = 14) or advanced (n = 16) stage of CRC patients. The horizontal lines indicate the median for individual groups. (B) The correlation between the percentages of CD14⁺CD169⁺ circulating monocytes and TIMs in early stage of CRC patients. (C) The correlation between the percentages of CD14⁺CD169⁺ circulating monocytes and TIMs in advanced stage of CRC patients. (D) The correlation between the levels of plasma CEA and the percentages of CD14⁺CD169⁺ circulating monocytes in the CRC patients. (E) The correlation between the levels of plasma CEA and the percentages of CD14⁺CD169⁺ TIMs in the CRC patients.</p

Data_Sheet_1_Phthalates and phthalate metabolites in urine from Tianjin and implications for platelet mitochondrial DNA methylation.docx

BackgroundPhthalates (PAEs) are important synthetic substances in plastics, attracting much attention due to their potential effects on the cardiovascular system.MethodsIn this study, urine and blood samples from 39 individuals were collected in Tianjin, China. Phthalates and phthalate metabolites (mPAEs) were analyzed using gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography-mass spectrometry (HPLC-MS), respectively. The polymerase chain reaction (PCR) products from bisulfite-treated mitochondrial DNA (mtDNA) samples were analyzed using pyrosequencing technology.ResultsThe detection frequencies for 9 PAEs varied from 2.56 to 92.31%, and those for 10 mPAEs varied from 30.77 to 100%. The estimated daily intakes (EDIs) and cumulative risk of PAEs were calculated based on the experimental statistics of urinary PAEs and mPAEs. For PAEs, the HIRfD (hazard index corresponding to reference doses) values of 10.26% of participants and the HITDI (hazard index corresponding to tolerable daily intake) values of 30.77% of participants were estimated to exceed 1, suggesting a relatively high exposure risk. The mtDNA methylation levels in the MT-ATP8 and MT-ND5 were observed to be lower than in the MT-ATP6. Mono-ethyl phthalate (MEP) and MT-ATP8 were positively correlated with triglyceride levels (p ConclusionThe effects of PAE exposure on cardiovascular diseases (CVDs) should be investigated further.</p

Characterization of CD14⁺CD169⁺IL-10⁺ M2 cells in CRC patients.

<p>PBMCs and TIMs were isolated from individual subjects and stimulated with LPS and PMA/ionomycin. The cells were stained with anti-CD169 and anti-CD14 for 30 min, fixed, and permeabilized, followed by intracellular staining with anti-IL-10 or anti-IL-12. The cells were first gated on CD14⁺CD169⁺ cells and the percentages of CD14⁺CD169⁺IL-12⁺ M1 and CD14⁺CD169⁺IL-10⁺ M2 cells in PBMCs or TIMs from individual subjects were determined by flow cytometry. The levels of plasma IL-10 and IL-12 in individual subjects were determined by ELISA. Data are representative charts or expressed as the values of individual patients. The horizontal lines indicate the median for individual groups. (A) Flow cytometry analysis. (B) The percentages of CD14⁺CD169⁺IL-10⁺ M2 cells. (C) The percentages of CD14⁺CD169⁺IL-12⁺ M1 cells. The horizontal lines indicate the median values. (D) The potential association between the percentages of circulating CD14⁺CD169⁺IL-10⁺ cells and CD14⁺CD169⁺ cells in CRC patients. (E) The potential association between the percentages of TIMs CD14⁺CD169⁺IL-10⁺ cells and CD14⁺CD169⁺cells in the CRC tissues. (F) The levels of plasma IL-10. (G) The levels of plasma IL-12. (H) The correlation of plasma IL-10 levels with the percentages of CD14⁺CD169⁺ IL-10⁺ monocytes in PBMCs from CRC patients. (I) The correlation between the levels of plasma IL-10 and CEA in CRC patients.</p