Characterization and application of aggregated porous copper oxide flakes for cupric source of copper electrodeposition

Copper oxide was prepared with thermal decomposition of basic copper carbonate to complement the concentration of cupric ions for copper electrodeposition in a plating system with insoluble anode. Copper oxide particles with a structure of aggregated porous flakes had a wide size distribution ranging from 100 nm to 100 μm. Copper oxide exhibited a dissolution rate of about 15 s in 12.5 vol% H2SO4 solution. During copper electrodeposition, copper deposits with fine growth formed in the electrolyte with stable cupric concentration provided by rapid dissolution of copper oxide

Aldose Reductase Is Involved in the Development of Murine Diet-Induced Nonalcoholic Steatohepatitis

<div><p>Hepatic aldose reductase (AR) expression is known to be induced in liver diseases, including hepatitis and hepatocellular carcinoma. However, the role of AR in the development of these diseases remains unclear. We performed this current study to determine whether and how AR might be involved in the development of diet-induced nonalcoholic steatohepatitis. Our results showed that the level of AR protein expression was significantly higher in db/db mice fed the methionine-choline-deficient (MCD) diet than in mice fed the control diet. In parallel with the elevation in AR, steatohepatitis was observed in MCD diet-fed mice, and this diet-induced steatohepatitis was significantly attenuated by lentiviral-mediated knock-down of the AR gene. This suppressive effect of AR knock-down was associated with repressed levels of serum alanine aminotransferase and hepatic lipoperoxides, reduced mRNA and protein expression of hepatic cytochrome P450 2E1 (CYP2E1), and decreased mRNA expression of pro-inflammatory tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Moreover, AR-induced elevations on the level of CYP2E1 expression, reactive oxygen species, mRNA expression of TNF-α and IL-6 were confirmed in AML12 hepatocytes. Further, lentiviral-mediated knock-down of AR ameliorated MCD diet-induced collagen deposition in the livers of db/db mice. With the improvement in liver fibrosis, the mRNA levels of tissue inhibitor of metalloproteinase-1 (TIMP-1) and matrix metalloproteinase-2 (MMP-2), two genes involved in hepatic fibrogenesis, were found to be significantly suppressed, while TIMP-2 and MMP-13 were unaffected. Together these data indicate that inhibition of AR alleviates the MCD diet-induced liver inflammation and fibrosis in db/db mice, probably through dampening CYP2E1 mediated-oxidative stress and ameliorating the expression of pro-inflammatory cytokines.</p></div

Effect of AR knock-down on scores for hepatic necroinflammation and fibrosis.

<p>The severity of hepatic necroinflammation and fibrosis were scored as described in the Materials and Methods. Values are means ± (<i>n</i> = 4/group).</p>*<p>P<0.05 compared with MCD+pLV-shNC.</p

Effect of lentiviral-mediated knock-down of AR on hepatic lipoperoxide content, CYP2E1 expression in db/db mice and over-expression of AR on CYP2E1 expression and ROS levels in AML12 cells.

<p>Hepatic lipoperoxide content (A), CYP2E1 mRNA (B) and protein (C) expression were determined in mice fed the control diet, MCD diet, or MCD diet treated with lentiviruses carrying shRNA against AR for 8 weeks (<i>n = </i>4). CYP2E1 protein expression (D) and ROS levels (E) were assayed in AML12 cells transfected with pFLAG-mAR for 36 hours (<i>n = </i>3). Values are expressed as the mean ± SEM. ***, P<0.001; **, P<0.01; *, P<0.05.</p

Effect of lentiviral-mediated knock-down of AR on MCD diet–induced collagen deposition in db/db mice.

<p>A. Masson’s trichrome–stained liver sections from mice fed (a) the control diet+pLV-shNC, (b) the MCD diet+pLV-shNC, and (c) the MCD diet+pLV-shAR. Long arrows point to perivenular and small arrows to pericellular fibrosis. Slides are representative of four separate experiments (original magnification, ×200). B. Total RNA was isolated and analyzed for collagen type I mRNA expression. Values are expressed as the mean ± SEM. *, P<0.05.</p

AR is involved in the development of MCD diet-induced steatohepatitis in db/db mice.

<p>A. Representative Western blot shows the induction of hepatic AR protein expression in MCD diet-fed mice. Average densitometric analyses of AR were calculated as fold increases over the control diet (<i>n = </i>4); values are expressed as the mean ± SEM. **, P<0.01. B. Hematoxylin and eosin–stained liver sections from mice fed: (a) Control diet+pLV-shNC. (b) MCD diet+pLV-shNC. (c) MCD diet+pLV-shAR. Arrows point to foci of necroinflammation. Slides are representative of four separate experiments (original magnification, ×100). C. Effect of MCD diet and lentiviral-mediated knock-down of AR on serum ALT levels in db/db mice. Data are means ± SEM of six mice in each group. ***, P<0.001; **, P<0.01.</p

Effect of AR on hepatic mRNA expression of proinflammatory cytokines.

<p>Total RNA was isolated and analyzed for TNF-α, IL-6 and TGF-β1 in db/db mice (A) fed the control or MCD diet and transduced with pLV-shNC or pLV-shAR for 8 weeks (<i>n = </i>4) and in AML12 cells (B) transfected with pFLAG-CMV2 or pFLAG-mAR for 36 hours (<i>n = </i>3). Values are expressed as the mean ± SEM. **, P<0.01; *, P<0.05.</p

Primer sequences used for amplification of mRNA by real-time PCR.

<p>Primer sequences used for amplification of mRNA by real-time PCR.</p