Primers and annealing temperatures (temp) for Notch and VEGF pathway molecules.

<p>Primers and annealing temperatures (temp) for Notch and VEGF pathway molecules.</p

Correlation analysis in missed abortion.

<p>A. the expression of VEGFR1 positively correlated with Notch1 in missed abortion; B. VEGFR2 expression positively correlated with Notch1 in missed abortion; C. Dll4 expression positively correlated with Notch1 in missed abortion; D. HIF-1a positively correlated with Notch1 in missed abortion; E. HIF-1a was closely negatively correlated with VEGF in missed abortion.</p

Novel Agent Nitidine Chloride Induces Erythroid Differentiation and Apoptosis in CML Cells through c-Myc-miRNAs Axis

<div><p>The proto-oncogene c-Myc plays critical roles in human malignancies including chronic myeloid leukemia (CML), suggesting that the discovery of specific agents targeting c-Myc would be extremely valuable for CML treatment. Nitidine Chloride (NC), a natural bioactive alkaloid, is suggested to possess anti-tumor effects. However, the function of NC in leukemia and the underlying molecular mechanisms have not been established. In this study, we found that NC induced erythroid differentiation, accompanied by increased expression of erythroid differentiation markers, e. g. α-, ε-, γ-globin, CD235a, CD71 and α-hemoglobin stabilizing protein (AHSP) in CML cells. We also observed that NC induced apoptosis and upregulated cleaved caspase-3 and Parp-1 in K562 cells. These effects were associated with concomitant attenuation of c-Myc. Our study showed that NC treatment in CML cells enhanced phosphorylation of Thr58 residue and subsequently accelerated degradation of c-Myc. A specific group of miRNAs, which had been reported to be activated by c-Myc, mediated biological functions of c-Myc. We found that most of these miRNAs, especially miR-17 and miR-20a showed strong decrement after NC treatment or c-Myc interference. Furthermore, overexpression of c-Myc or miR-17/20a alleviated NC induced differentiation and apoptosis in K562 cells. More importantly, NC enhanced the effects of imatinib in K562 and primary CML cells. We further found that even imatinib resistant CML cell line (K562/G01) and CML primary cells exhibited high sensitivity to NC, which showed potential possibility to overcome imatinib resistance. Taken together, our results clearly suggested that NC promoted erythroid differentiation and apoptosis through c-Myc-miRNAs regulatory axis, providing potential possibility to overcome imatinib resistance.</p></div

Correlation analysis in induced abortion.

<p>A.VEGF positively correlated withVEGFR1. B.VEGF expression close positively correlated with VEGFR2. C.VEGFR1 expression close positively correlated with VEGFR2. D-,G.Dll4 positively correlated with VEGF,VEGFR1, VEGFR2, Notch1. H,I. HIF-1a positively correlated with VEGFR2 or Dll4.</p

VEGF and Notch pathway molecules RNA levels in missed abortion and induced abortion.

<p>VEGF and Notch pathway molecules RNA levels in missed abortion and induced abortion.</p

The levels of circulation Tc17 cells in representative controls, CIN patients and UCC patients.

<p>Upper right quadrants are the domains of Tc17 (CD8⁺ IL-17⁺) cells and the percentages of them were shown in each panel.</p

Clinical Characteristics of UCC Patients.

<p>Abbreviation: FIGO, International Federation of Gynecologists and Obstetricians; SCC, squamous cell carcinoma; ADC, adenocarcinoma; ADSC, adenosquamous carcinoma.</p

The correlations between Tc17 cells and Th17 cells.

<p>(a, b) Linear regression analysis between frequencies of Tc17 cell and Th17 cells in the blood (r_CIN = 0.435, <i>P</i> = 0.042, n = 22; r_UCC = 0.403, <i>P</i> = 0.016, n = 36). (c, d) Linear regression analysis between the levels of Tc17 cells and Th17 cells in the cervical tissues (r_CIN = 0.441, <i>P</i> = 0.039, n = 17; r_UCC = 0.693, <i>P</i> = 0.026, n = 30).</p

The correlations between Tc17 cells and microvessel density (MVD).

<p>(A) Linear regression analysis between the levels of Tc17 cells and MVD (Total: r = 0.987, <i>P</i><0.0001, n = 65; control: r = 0.814, <i>P</i><0.001, n = 18, CIN: r = 0.923, <i>P</i><0.001, n = 17; UCC: r = 0.938, <i>P</i><0.0001, n = 30). MVD from each sample was plotted against Tc17 cells level from the same person. (B) Representative immunohistochemical staining of MVD in cervical tissues of three groups. Representative sites with low (100×, upper panels) and high (400×, lower panels) magnification were shown.</p

Expression of Tc17 cells in cervical tissues of the three groups.

<p>(A) Immunohistochemical double staining for Tc17 cells in the control group (a and d), CIN group (b and e) and UCC group (c and f). Representative sites with low (200×, upper panels) and high (400×, lower panels) magnification were shown. (B) IL-17-producing cells were stained red (in the cytoplasm) and CD8⁺ cells were stained black (in the membrane). The co-expression of CD8 and IL-17 confirmed that a proportion of Tc17 cells.</p