The effect of ghrelin on HF.

<p>(A) Representative images of hearts. Four weeks after ghrelin administration, the heart was removed, and the tissue sections were stained with H&E (×200 magnification). (B) Hemodynamic index, including ±dp/dt_max, LVEDP, and LVSP. +dp/dt_max, maximal rate of the rise in blood pressure in the ventricular chamber; -dp/dt_max, maximal rate of the decline in blood pressure in the ventricular chamber; LVEDP, left ventricular end diastolic pressure; LVSP, left ventricular systolic pressure. (C) The levels of plasma BNP. The data are presented as the means ± SD. <b>**</b>P<0.01 vs. SO group; ^#P<0.05, ^##P<0.01 vs. MI group.</p

Ghrelin inhibited cardiomyocyte apoptosis both in vivo and in vitro.

<p>(A) TUNEL analysis was performed after the end of the ghrelin treatment. The TUNEL-positive cells (apoptotic cells) are indicated by arrows. The data are presented as the means ± SD. <b>**</b>P<0.01 vs. SO group, ^##P<0.01 vs. MI group. (B) Ang II-induced DNA fragmentation in cardiomyocytes with or without ghrelin. Cultured cardiomyocytes from neonatal rats were stimulated with or without Ang II and ghrelin for 24 hours. The cardiomyocyte lysate was first incubated with RNase and then with proteinase K. By this method, only fragmented DNA was extracted. The DNA was separated by electrophoresis on a 1.5% agarose gel and stained with ethidium bromide. Lane A, vehicle-treated cardiomyocytes; lane B, cardiomyocytes incubated with 0.1 µmol/L Ang II; lane C, cardiomyocytes incubated with 1 µmol/L Ang II; lane D, cardiomyocytes incubated with 0.1 µmol/L ghrelin and 0.1 µmol/L Ang II; lane E, cardiomyocytes incubated with 0.1 µmol/L ghrelin. (C) Apoptosis of cardiomyocytes treated with Ang II and ghrelin. Cardiomyocytes were incubated in culture medium (control), 0.1 µmol/L Ang II, 0.1 µmol/L ghrelin, or 0.1 µmol/L ghrelin + 0.1 µmol/L Ang II for 24 hours. Magnification: ×200. The graph presents the percentage of TUNEL-positive cells determined from 100 cells in 3 independent experiments. The data are presented as the means ± SD. **P<0.01 vs. the control group;^##P<0.01 vs. the Ang II group.</p

Real-time quantitative PCR was performed to determine the AT2 receptor mRNA levels.

<p>After the addition of 0.1 µmol/L ghrelin, 0.1 µmol/L Ang II, 0.1 µmol/L ghrelin + 0.1 µmol/L Ang II, or culture medium (control), the cardiomyocytes were cultured for 24 hours. Real-time quantitative PCR was performed to determine the mRNA levels. The data are presented as the means ± SD. *P<0.05 vs. the control group.</p

Primer sequences used in real-time PCR.

<p>Primer sequences used in real-time PCR.</p

Ghrelin mediated myocardium protection by down-regulating caspase-3.

<p>(A) The expression of caspase-3 mRNA by real-time quantitative PCR. The total RNA of noninfarcted left ventricular tissue was extracted using TRIzol Reagent, and real-time quantitative PCR was performed to determine the caspase-3 mRNA levels. The data are presented as the means ± SD. **P<0.01 vs. SO group, ^##P<0.01 vs. MI group. (B) The expression of caspase-3 in rat cardiac tissues examined by immunohistochemical staining (×200). (C) Analysis of caspase-3 mRNA levels in cardiomyocytes by quantitative real-time RT-PCR. After the addition of 0.1 µmol/L ghrelin, 0.1 µmol/L Ang II, 0.1 µmol/L ghrelin + 0.1 µmol/L Ang II, or culture medium (control), the cardiomyocytes were cultured for 24 hours. Real-time quantitative PCR was performed to determine the mRNA levels. The data are presented as the means ± SD. **P<0.01 vs. control group; ^##P<0.01 vs. Ang II group. (D) Detection of caspase-3 protein expression in rat cardiomyocytes by immunocytochemical staining. After the addition of 0.1 µmol/L ghrelin, 0.1 µmol/L Ang II, 0.1 µmol/L ghrelin + 0.1 µmol/L Ang II, or culture medium (control), the cardiomyocytes were cultured for 24 hours. Magnification: ×200.</p

Ghrelin inhibits Ang II-induced cell apoptosis by down-regulating AT1R and thereby preventing HF.

<p>(A) The expression of AT1R mRNA by real-time quantitative PCR. The total RNA of noninfarcted left ventricular tissue was extracted using TRIzol Reagent, and real-time quantitative PCR was performed to determine the AT1R mRNA levels. The data are presented as the means ± SD. **P<0.01 vs. SO group, ^##P<0.01 vs. MI group. (B) The expression of cardiac AT1R in rat cardiac tissues by immunohistochemical staining (×200). (C) Real-time quantitative PCR was performed to determine the AT1 receptor mRNA levels in cardiomyocytes. After the addition of 0.1 µmol/L ghrelin, 0.1 µmol/L Ang II, 0.1 µmol/L ghrelin + 0.1 µmol/L Ang II, or culture medium (control), the cardiomyocytes were cultured for 24 hours. Real-time quantitative PCR was performed to determine the mRNA levels. The data are presented as the means ± SD. **P<0.01 vs. the control group; ^##P<0.01 vs. the Ang II group. (D) Western blotting was performed to determine AT1 receptor expression. Proteins were extracted from cardiomyocytes, separated by SDS-PAGE, and immunoblotted sequentially with anti-AT1 receptor antibody. The graph shows the result of densitometric quantification of the AT1 receptor protein relative to GAPDH as an internal control. The data are presented as the means ± SD. *P<0.05, **P<0.01 vs. the control group; ^##P<0.01 vs. the Ang II group.</p

Characteristics of patients with CHF and the control group.

<p>Characteristics of patients with CHF and the control group.</p

A novel phenolic acid from the fruits of <i>Rosa soulieana</i>

<div><p>From the <i>n</i>-BuOH-soluble fraction of a MeOH extract of the fruits of <i>Rosa soulieana</i>, one new phenolic glucoside (<b>1</b>) was isolated along with five known compounds, comprising two lignin glycosides, two flavonoid glycosides and a phenolic glycoside. The chemical structure of the new compound was elucidated by extensive spectroscopic analyses, including ESI-MS, UV, IR, ¹H and ¹³C NMR, DEPT and 2D NMR (HSQC and HMBC). All the isolated compounds were evaluated for their antioxidant activity by using ABTS (2,2′-azino-bis(3-ethylbenzoline-6-sulfonic acid)) assay. Among these compounds, <b>1</b>, <b>3</b> and <b>6</b> exhibited strong scavenging activity in ABTS^√+(SC₅₀ = 102.10, 193.85, 65.38 μmol/L, respectively) compared with the positive control l-ascorbic acid (<i>Vc</i>) (SC₅₀ = 117.16 μmol/L).</p></div

Sequences after initial processing in QIIME (seqs.fna, library 1)

Raw sequence data from the Roche 454 GS FLX sequencer, region 1 (split_library_output_1). These data are the output of the command: split_libraries.py -m 454_Map.txt -f 1.TCA.454Reads.fna -q 1.TCA.454Reads.qual -o split_library_output_1/ -l 100 -L 700 -H 9 -M 2 -b 1

MiR-541-5p regulates lung fibrosis by targeting cyclic nucleotide phosphodiesterase 1A

<p><b>Aim of the Study:</b> Idiopathic pulmonary fibrosis (IPF) is a lethal human disease with short survival time and few treatment options. In this study, we aim to demonstrate that cyclic nucleotide phosphodiesterase 1A (PDE1A), a Ca2+/calmodulin-stimulating PDE family member, plays a critical role in the induction of fibrosis and angiogenesis in the lung. <b>Materials and Methods:</b> To induce pulmonary damage, adult male SD rats were treated with bleomycin in a dose of 6 mg/kg body weight by a single intratracheal instillation. For in vivo silencing of PDE1A in rat lung, a nonspecific control siRNA or PDE1A-specific siRNA was used to treat rat through nasal instillation. Human normal pulmonary fibroblasts MRC-5 and hFL1 and rat lung fibroblasts were used as in vitro model. Immunohistochemistry and immunoflurescence staining were performed to detect PDE1A and α-SMA expression. Reverse transcription-qPCR was performed to detect microRNA and mRNA expression. In vitro wound healing assay was performed to detect pulmonary fibroblasts'mortality ability. <b>Results:</b> In vitro studies showed that PDE1A can stimulate lung fibroblasts to undergo myofibroblastic changes. This led to the identification of miR-541-5p as one of the miRNA candidates associated with bleomycin response. We found that miR-541-5p expression is downregulated in TGF-β-treated lung fibroblasts and the rat pulmonary fibrosis model. Overexpression of miR-541-5p in lung fibroblasts inhibited mortality of human lung fibroblasts. <b>Conclusions:</b> MiR-541-5p is a key effector in lung fibroblastsby by regulating PDE1A expression at protein translation level and its overexpression is protective against bleomycin-induced lung fibrosis.</p