Synthesis and Self-Assembly of Rod–Rod Block Copolymers with Different Rod Diameters

We synthesized a series of rod–rod block copolymers, poly­(octyl-4′-(octyloxy)-2-vinylbiphenyl-4-carboxylate)-<i>b</i>-poly­(γ-benzyl-_l-glutamate) (PVBP-<i>b</i>-PBLG) with different rod diameters, and we focused on the effect of size disparity on their self-assembling behaviors in bulk. By using differential scanning calorimetry, wide-angle X-ray diffraction (WAXD), small-angle X-ray scattering, and transmission electron microscopy techniques, we found that the columnar nematic (Φ_N) liquid crystalline phase of PVBP and the hexagonal columnar (Φ_H) phase of PBLG were disrupted to different extents depending on the volume fractions of PVBP (<i>f</i>_PVBP’s). When <i>f</i>_PVBP is about 60%, the Φ_H phase of PBLG is retained with the appearance of the two second-order diffractions in WAXD patterns. When <i>f</i>_PVBP is increased to above 73%, the ordered structure of PBLG is destroyed, and when <i>f</i>_PVBP is 91% even the first-order diffraction peak disappears. At the same time, the Φ_N phase of PVBP changes from poorly ordered to well ordered with increasing <i>f</i>_PVBP. Accordingly, with <i>f</i>_PVBP increasing from 60% to 91%, the microphase-separated structure goes through a poorly ordered structure to an interdigitated lamellar morphology and to a bilayer lamellar structure

HGF and Direct Mesenchymal Stem Cells Contact Synergize to Inhibit Hepatic Stellate Cells Activation through TLR4/NF-kB Pathway

<div><h3>Aims</h3><p>Bone marrow-derived mesenchymal stem cells (BMSCs) can reduce liver fibrosis. Apart from the paracrine mechanism by which the antifibrotic effects of BMSCs inhibit activated hepatic stellate cells (HSCs), the effects of direct interplay and juxtacrine signaling between the two cell types are poorly understood. The purpose of this study was to explore the underlying mechanisms by which BMSCs modulate the function of activated HSCs.</p> <h3>Methods</h3><p>We used BMSCs directly and indirectly co-culture system with HSCs to evaluate the anti-fibrosis effect of BMSCs. Cell proliferation and activation were examined in the presence of BMSCs and HGF. c-met was knockdown in HSCs to evaluate the effect of HGF secreted by BMSCs. The TLR4 and Myeloid differentiation primary response gene 88(MyD88) mRNA levels and the NF-kB pathway activation were determined by real-time PCR and western blotting analyses. The effect of BMSCs on HSCs activation was investigated in vitro in either MyD88 silencing or overexpression in HSCs. Liver fibrosis in rats fed CCl₄ with and without BMSCs supplementation was compared. Histopathological examinations and serum biochemical tests were compared between the two groups.</p> <h3>Results</h3><p>BMSCs remarkably inhibited the proliferation and activation of HSCs by interfering with LPS-TLR4 pathway through a cell–cell contact mode that was partially mediated by HGF secretion. The NF-kB pathway is involved in HSCs activation inhibition by BMSCs. MyD88 over expression reduced the BMSC inhibition of NF-kB luciferase activation. BMSCs protected liver fibrosis in vivo.</p> <h3>Conclusion</h3><p>BMSCs modulate HSCs in vitro via TLR4/MyD88/NF-kB signaling pathway through cell–cell contact and secreting HGF. BMSCs have therapeutic effects on cirrhosis rats. Our results provide new insights into the treatment of hepatic fibrosis with BMSCs.</p> </div

Primary Cells morphology and molecular expression.

<p>(A) HSC cell line LX2 (left) and GFP labeled LX2 (right). (B) BMSCs were separated from bone marrow and purified by removing the non-adherent cells at least three times. The 3rd generation BMSCs (medium) were fusiform and fibroblast-like (100×). Primary human HSC (right). (C) FCM analysis showed that the 3rd generation BMSCs expressed CD44 and CD90 but not CD45 or CD34.</p

BMSC inhibited LPS induced activation of LX2 and upregulation of TLR4.

<p>(A) The effect of different dose of LPS on LX2 cells activation as the IL-8 and TGF-β secretion. All data are shown as mean ± SD from three independent experiments. * P<0.05. (B) Relative mRNA expression of TLR4 in LX2 cells and two HSCs in response to LPS. Data are depicted relative to expression in cells without LPS, which are assigned a value of 1. (C) RT-PCR analysis of the HSC activation marker α-SMA, Col-1 and TLR4 in the indicated treatments. (D) Western blot analysis of the HSC activation marker α-SMA, Col-1 and TLR4 in the indicated treatments. Expression levels were normalized with GAPDH.</p

HGF is the major cytokine involved in BMSC-mediated inhibition of LX2 activation.

<p>(A) Relative mRNA expression of Col-1 and α-SMA in LX2 with different cytokine stimulation. Data are depicted relative to expression in LX2 cells cultured with PBS, which are assigned a value of 1. All data are shown as mean ± SD from three independent experiments. * P<0.05. (B) Western blot analysis for TLR4 expression under different combination of cytokine. GAPDH was used as a loading control for all Western blots (C) Knockdown of c-met in two specific shRNA-transduced LX2 cell lines. (D) Western blot analysis for Col-1 and α-SMA expression in LX2 with c-met knockdown.</p

MACC1 as a Prognostic Biomarker for Early-Stage and AFP-Normal Hepatocellular Carcinoma

<div><p>Background</p><p>The metastasis-associated in colon cancer 1 gene (MACC1) has been found to be associated with cancer development and progression. The aim of this study was to investigate the prognostic value of MACC1 in early-stage and AFP-normal hepatocellular carcinoma (HCC).</p><p>Methods</p><p>mRNA and protein levels of MACC1 expression in one normal liver epithelial cells THLE3 and 15 HCC cell lines were examined using reverse transcription-PCR and Western blot. MACC1 expression was also comparatively studied in 6 paired HCC lesions and the adjacent non-cancerous tissue samples. Immunohistochemistry was employed to analyze MACC1 expression in 308 clinicopathologically characterized HCC cases. Statistical analyses were applied to derive association between MACC1 expression scores and clinical staging as well as patient survival.</p><p>Results</p><p>Levels of MACC1 mRNA and protein were higher in HCC cell lines and HCC lesions than in normal liver epithelial cells and the paired adjacent noncancerous tissues. Significant difference in MACC1 expression was found in patients of different TNM stages (<i>P</i><0.001). Overall survival analysis showed that high MACC1 expression level correlated with lower survival rate (<i>P</i> = 0.001). Importantly, an inverse correlation between MACC1 level and patient survival remained significant in subjects with early-stage HCC or with normal serum AFP level.</p><p>Conclusions</p><p>MACC1 protein may represent a promising biomarker for predicting the prognosis of HCC, including in early-stage and AFP-normal patients.</p></div

MyD88 was necessary for BMSCs to protect LPS induced activation of LX2.

<p>(A) Western blotting analysis of MyD88 expression in indicated cells. GAPDH was used as a loading control. (B) TGF-β, a-SMA, Col-1 and TLR4 was analysis by western blotting on LPS stimulated LX2 cell under BMSC treatment, MyD88 overexpress (MyD88) and MyD88 RNAi#1. (C) Relative NF-kB luciferase activity in the LPS stimulated LX2 cell with BMSC treated and MyD88 overexpress and MyD88 shRNA. (D) Proposed scheme for BMSC inhibition in LPS-stimulated signaling in the activation of LX2.</p

[Vala Nurettin'in babası Nurettin Bey'e ait fotoğraf)

Taha Toros Arşivi, Dosya No: 8-Va-Nu (Vala Nurettin)Unutma İstanbul projesi İstanbul Kalkınma Ajansı'nın 2016 yılı "Yenilikçi ve Yaratıcı İstanbul Mali Destek Programı" kapsamında desteklenmiştir. Proje No: TR10/16/YNY/010

Silencing URGCP/URG4 expression impaired cell proliferation through FOXO3a.

<p>Western blotting analysis of p21^Cip1, p27^Kip1, cyclin D1 and FOXO3a proteins in indicated Hep3B (A) and QGY-7703 (B) cell lines. Silencing FOXO3a increased the proliferation of URGCP/URG4 shRNA(s)-transduced cells as determined by MTT assay in Hep3B (C) and QGY-7703 (D) cells. GAPDH was used as a loading control for all Western blots. Error bars represent SD from three independent experiments. *<i>P</i><0.05.</p