DIM prevents neointima formation induced by guidewire injury.

<p>A–B. Representative sections of the injured carotid artery of either an animal from the control group or the DIM-treated group are shown. C–D. Quantification of the intimal area and I/M ratios of carotid arteries of mice from either the control group or the DIM-treated group (n = 6, *<i>P</i><0.01 versus injured control). E. Quantification of PCNA-positive cells of carotid arteries of mice from either the control group or the DIM-treated group (n = 9; *<i>P</i><0.01 versus injured control).</p

Effect of DIM on reendothelialization, inflammation, apoptosis, and extracellular matrix deposition in vivo.

<p>A. Representative immunohistochemical staining for CD31 at day 7 and day 28 after injury. B, Quantitative analysis showed no difference in the extent of reendothelialization between the 2 groups (n = 6, <i>P</i> = NS versus injured control, *<i>P</i><0.05 versus day 7). C. Infiltration of inflammatory cells at 7days after vascular injury. Inflammatory cells were immunostained with anti-CD45 antibody. Arrows indicate positive cells (n = 6, *<i>P</i><0.05 versus injured control). D. Apoptotic cells were assessed by TUNEL method at 7 days after injury. Arrows indicate TUNEL-positive cells. E. Sirius red staining of injured vessels (28 days). Collagen fibers were stained in red.</p

Damage-Free Back Channel Wet-Etch Process in Amorphous Indium–Zinc-Oxide Thin-Film Transistors Using a Carbon-Nanofilm Barrier Layer

Amorphous indium–zinc-oxide thin film transistors (IZO-TFTs) with damage-free back channel wet-etch (BCE) process were investigated. A carbon (C) nanofilm was inserted into the interface between IZO layer and source/drain (S/D) electrodes as a barrier layer. Transmittance electron microscope images revealed that the 3 nm-thick C nanofilm exhibited a good corrosion resistance to a commonly used H₃PO₄-based etchant and could be easily eliminated. The TFT device with a 3 nm-thick C barrier layer showed a saturated field effect mobility of 14.4 cm² V^–1 s^–1, a subthreshold swing of 0.21 V/decade, an on-to-off current ratio of 8.3 × 10¹⁰, and a threshold voltage of 2.0 V. The favorable electrical performance of this kind of IZO-TFTs was due to the protection of the inserted C to IZO layer in the back-channel-etch process. Moreover, the low contact resistance of the devices was proved to be due to the graphitization of the C nanofilms after annealing. In addition, the hysteresis and thermal stress testing confirmed that the usage of C barrier nanofilms is an effective method to fabricate the damage-free BCE-type devices with high reliability

DIM prevents VSMC proliferation and DNA synthesis induced by PDGF-BB.

<p>VSMCs were serum starved for 24 h and then treated with the indicated concentrations of DIM (5 to 50 µM) for 48 h in the absence or presence of PDGF-BB (20 ng/ml). A. Cell viability was examined with the WST-1 test. The data are expressed as the mean OD450±SEM (#<i>P</i><0.01 versus control group; *<i>P</i><0.01 versus PDGF alone; n = 6). B. BrdU incorporation was determined with an ELISA-based assay. DNA synthesis is expressed as the mean OD370±SEM (#<i>P</i><0.01 versus control group; *<i>P</i><0.01 versus PDGF alone; n = 6).</p

DIM prevents cell cycle progression in VSMCs.

<p>VSMCs were grown with DIM (25 µM) in the absence or presence of PDGF-BB (20 ng/ml) for 24 h, and cell cycle distribution was evaluated with flow cytometric analysis. A. Representative cell cycle profiles are shown. B. Quantification of VSMCs in the G0/G1, S, and G2/M phases, as determined by flow cytometric evaluation, is shown (#<i>P</i><0.01 versus control group; *<i>P</i><0.01 versus PDGF alone; n = 3). C. Cell cycle protein expression was measured with western blot analysis. GAPDH detection served as a loading control.</p

Inhibitory effects of DIM on PDGF-Rβ, Akt, GSK-3β, ERK1/2, and STAT3 activation in the PDGF-BB-stimulated VSMCs.

<p>Serum-starved VSMCs were stimulated with PDGF-BB for an indicated time in the absence or presence of DIM (25 µM). The protein levels of phospho-PDGF-Rβ, PDGF-Rβ, phospho-Akt, Akt, phospho-GSK-3β, GSK-3β, phospho-ERK1/2, ERK1/2, phospho-STAT3, and STAT3 were determined with western blot analysis. One representative image out of 3 independently performed experiments is shown.</p

Effect of DIM on the regulation of smooth muscle gene expression.

<p>A. VSMCs preincubated for 2 h with DIM and then stimulated with PDGF-BB (20 ng/ml) for 48 h. The protein levels of SM α-actin, SM22α, and desmin were determined with western blot analysis and quantified by densitometry. B. Bar graphs showing the quantification of western blots. The results are expressed as a percentage of control (#<i>P</i><0.05 versus control group; *<i>P</i><0.05 versus PDGF alone; n = 3).</p

Effect of DIM on viability of VSMC and HUVEC.

<p>A. VSMCs were incubated in growth medium in the absence or presence of different concentrations of DIM for 24 hours, and cell viability was evaluated by counting the number of cells that excluded the trypan blue dye (<i>P</i> = NS versus control group; n = 4). B, HUVECs were incubated in growth medium in the absence or presence of different concentrations of DIM for 24 hours, and cell viability was evaluated by trypan blue exclusion (<i>P</i> = NS versus control group; n = 4). C, VSMCs were incubated in the absence or presence of DIM (25 µM) or H₂0₂ (400 µM, serving as positive control) for 24 hours. After 3 washing steps, cells were then stimulated with PDGF-BB for 24 hours, and VSMC proliferation was quantified by BrdU incorporation (<i>P</i> = NS versus pretreated with control buffer; n = 6).</p

DIM inhibits PDGF-BB-induced cell migration.

<p>A. VSMCs were cultured in a cell migration filter insert and stimulated with PDGF-BB for 6 h with or without DIM treatment (25 µM). B. Cellular migration was determined by counting the cells that migrated through the pores. The results are expressed as means±SEM (#<i>P</i><0.01 versus control group; *<i>P</i><0.01 versus PDGF alone).</p

Optimization and Synthesis of Pyridazinone Derivatives as Novel Inhibitors of Hepatitis B Virus by Inducing Genome-free Capsid Formation

The capsid of hepatitis B virus (HBV) plays a vital role in virus DNA replication. Targeting nucleocapsid function has been demonstrated as an effective approach for anti-HBV drug development. A high-throughput screening and mechanism study revealed the hit compound <b>4a</b> as an HBV assembly effector (AEf), which could inhibit HBV replication by inducing the formation of HBV DNA-free capsids. The subsequent SAR study and drug-like optimization resulted in the discovery of the lead candidate <b>4r</b>, with potent antiviral activity (IC₅₀ = 0.087 ± 0.002 μM), low cytotoxicity (CC₅₀ = 90.6 ± 2.06 μM), sensitivity to nucleoside analogue-resistant HBV mutants, and synergistic effect with nucleoside analogues in HepG2.2.15 cells