Dysregulation of HULC promotes contrast-induced nephropathy (CIN) via regulating signaling pathway of miRNA-512 and prostaglandin E1 (PGE1).

It has been shown that contrast-induced nephropathy (CIN) can be attenuated by the administration of PGE1. As an enzyme responsible for the production of PGE1, PTGS1 was confirmed in this study as a miR-512 target. Meanwhile, HULC has been identified as a competing endogenous RNA of miR-512. Therefore, in this study, we tested the diagnostic value of HULC and miR-512 in subjects with or without CIN. In addition, we evaluated the regulatory relationship among HULC, miR-512, PTGS1 and PGE1 in vitro. We enrolled 320 patients with coronary heart disease and divided them into a CIN group and a non-CIN group. Subsequently, we detected the differential expression of miR-512, HULC and PGE1 in the two groups. We also used a dual luciferase reporter assay to evaluate the regulatory relationship among HULC, miR-512, PTGS1 and PGE1 in THP-1 cells. In patients with CIN, the expression levels of HULC and PGE1 were lower, but the expression level of miR-512 was higher. MiR-512 could directly bind to and negatively regulate the expression of PTGS1 and HULC. The expression of HULC was positively correlated with the expression of PTGS1 and PGE1, while negatively correlated with the expression of miR-512. The findings of this study demonstrated that deregulation of lncRNA-HULC/miR-512/PTGS1/PGE1 might be involved in the pathogenesis of CIN.

High COL4A3 expression correlates with poor prognosis after cisplatin plus gemcitabine chemotherapy in non-small cell lung cancer.

In this study, we investigated whether COL4A3 mRNA expression levels were associated with clinical outcomes after treatment with a combination of gemcitabine (Gem)/cis-diamminedichloroplatinum(II) (CDDP) regimen for patients with advanced stage non-small cell lung cancer (NSCLC). Response and survival were correlated with the level of COL4A3 expression in 58 patients with advanced (stage IIIb or IV) NSCLC treated as part of a multicenter randomized trial with Gem 1,250 mg/m(2) on days 1 and 8 plus CDDP 100 mg/m(2) on day 1 every 3 weeks. mRNA was isolated from paraffin-embedded pretreatment primary tumor specimens, and relative expression levels of COL4A3/ß-actin were measured using a quantitative reverse transcription-PCR (Taqman) system. COL4A3 expression was detectable in all tumors. There were no significant differences in COL4A3 levels by gender, age, performance status, weight loss, or tumor stage. The overall response rate was 45.8 %. There were no significant associations between COL4A3 expression and response. Median overall survival was significantly longer in patients with low COL4A3 expression tumors compared to patients with high expression tumors. COL4A3 expression, Eastern Cooperative Oncology Group performance status, and presence of weight loss were significant prognostic factors for survival in a Cox proportional hazards multivariable analysis. These data suggest that COL4A3 expression is a predictive factor for survival after CDDP/Gem therapy in advanced NSCLC. Although there was a trend toward decreased response with high COL4A3 mRNA levels, this difference failed to reach statistical significance. This result may reflect the impact of Gem and the requirement for COL4A3 expression for CDDP/Gem synergism or may be attributable to the relatively small patient sample size in this study. Prospective studies of COL4A3 as a predictive marker for activity of CDDP-based regimens in NSCLC are warranted.

Atomistic Modeling of the Negative Thermal Expansion in δ- Plutonium Based on the Two-State Description.

The δ phase of plutonium with the fcc structure exhibits an unusual negative thermal expansion (NTE) over its narrow temperature range of stability, 593-736 K. An accurate description of the anomalous high-temperature volume effect of plutonium goes beyond the current capability of electronic-structure calculations. We propose an atomistic scheme to model the thermodynamic properties of δ-Pu based on the two-state model of Weiss for the Invar alloys, inspired by the simple free-energy analysis previously conducted by Lawson et al. The two-state mechanism is incorporated into the atomistic description of a many-body interacting system. Two modified embedded atom method potentials are employed to represent the binding energies of two competing electronic states in δ-Pu. We demonstrate how the NTE takes place in δ-Pu by means of Monte Carlo simulations implemented with the two-state mechanism.

Tumor necrosis factor-alpha downregulates endothelial nitric oxide synthase mRNA stability via translation elongation factor 1-alpha 1.

Endothelium-derived nitric oxide (NO) is an important regulator of vascular function. NO is produced by endothelial NO synthase (eNOS), whose expression is downregulated by tumor necrosis factor (TNF)-alpha at the posttranscriptional level. To elucidate the molecular basis of TNF-alpha-mediated eNOS mRNA instability, eNOS 3' untranslated region (3'-UTR) binding proteins were purified by RNA affinity chromatography from cytosolic fractions of TNF-alpha-stimulated human umbilical vein endothelial cells (HUVECs). The formation of 3'-UTR ribonucleoprotein complexes, with molecular weight of 52 and 57 kDa, was increased by TNF-alpha. Matrix-assisted laser desorption ionization time-of-flight mass spectrometric analysis of the 52-kDa protein identified 3 peptides that comprise the peptide sequence of translation elongation factor 1-alpha 1 (eEF1A1). In HUVECs, TNF-alpha rapidly increased eEF1A1 expression, which is maximal after 1 hour and persists for up to 48 hours. RNA gel mobility-shift and UV cross-linking assays indicated that recombinant glutathione S-transferase-eEF1A1 fusion protein specifically binds to a UC-rich sequence in the 3'-UTR of eNOS mRNA. In addition, the domain III of eEF1A1 mediates the binding of eNOS 3'-UTR in eEF1A1. Overexpression of eEF1A1 markedly attenuated the expression of eNOS and luciferase gene fused with eNOS 3'-UTR in both COS-7 cells and bovine aortic endothelial cells (BAECs). Furthermore, adenovirus-mediated overexpression of eEF1A1 increased eNOS mRNA instability, whereas knockdown of eEF1A1 substantially attenuated TNF-alpha-induced destabilization of eNOS mRNA and downregulation of eNOS expression in HUVECs. These results indicate that eEF1A1 is a novel eNOS 3'-UTR binding protein that plays a critical role in mediating TNF-alpha-induced decrease in eNOS mRNA stability.

Circulating preprandial ghrelin to obestatin ratio is increased in human obesity.

CONTEXT: Obestatin, a sibling of ghrelin derived from preproghrelin, opposes ghrelin's effects on food intake. Plasma obestatin profiles in relation to ghrelin have not been fully investigated in human obesity. OBJECTIVE: We hypothesize that obesity might present with imbalance of circulating ghrelin and obestatin levels. PARTICIPANTS AND SETTING: Sixteen obese (eight men, aged 58.8 +/- 4.9 yr; eight women, aged 59.9 +/- 9.6 yr) and 14 normal-weight individuals (seven men, aged 52.7 +/- 5.9 yr; seven women, aged 56.1 +/- 4.9 yr) were evaluated at the in-patient department of Changhai Hospital, Shanghai, China. MAIN OUTCOME MEASURES: Total plasma ghrelin and obestatin levels, 1 h before and 2 h after breakfast, were measured by RIA. RESULTS: Both preprandial plasma ghrelin levels (P < 0.01) and obestatin levels (P < 0.01) were lower in the obese compared with normal-weight controls. However, unexpectedly, the ratio of preprandial ghrelin to obestatin was higher in obese compared with normal-weight controls (P < 0.01) even after adjustment for gender and age (P < 0.01). The ratio of postprandial ghrelin to obestatin was decreased both in obese (P < 0.05) and controls (P < 0.01) compared with their preprandial levels. There were no significant differences in the ratio of postprandial ghrelin to obestatin between obese and normal-weight controls. Body mass index was positively correlated with and was a significantly independent determinant of the preprandial ghrelin to obestatin ratio. CONCLUSION: Circulating preprandial ghrelin to obestatin ratio is elevated in human obesity. We suggest that high preprandial ghrelin to obestatin ratio may be involved in the etiology and pathophysiology of obesity.

[Effects of fusion gene encoding the hVEGF165 and fused hirudin on restenosis of injured carotid artery induced by angioplasty].

OBJECTIVE: To study the effects of fusion gene encoding the hVEGF(165) and fused hirudin on restenosis of injured carotid artery. METHODS: A fusion gene encoding hVEGF(165) and fused hirudin (hVEGF(165)-FH) was constructed and clone into the eukaryotic expression vector pcDNA3.0, thus constructing the plasmid VEGF(165)-FH/pcDNA3.0. Its activities to stimulate endothelial cell proliferation and to inhibit thrombosis were identified. Sixteen New Zealand rabbits underwent ligation of external carotid artery and a balloon was inserted into the common carotid artery for 30 minutes so as to construct model of restenosis of injured carotid artery. Then the rabbits were randomly divided into 4 equal groups. In the one-week and 3-week control groups, 400 microg of DNA of the plasmid pcDNA3.0 were transfused into the arterial lumen at the injured part immediately after the angioplasty, and 400 microg of DNA of the plasmid VEGF(165)-FH/pcDNA3.0 were transfused in the 1-week and 3-week experimental groups. One week and 3 weeks after the treatment peripheral blood samples were collected to detect the activated partial thromboplastin time (APTT), thrombin time (TT), and platelet aggregation rate, and then the rabbits underwent angiography to observe the situation of restenosis. Then the rabbits were killed to take out the injured part of artery to undergo pathological examination and Western blotting. RESULTS: The values of APTT, TT, and platelet aggregation rate were not significantly different among the 4 groups. Angiography conducted 1 and 3 weeks later showed that restenosis was significantly mild in the 2 experimental groups in comparison with the 2 control groups, and severe restenosis was seen in the 3-week control group. Western blotting showed that expression of specific fused protein could be found in the 1-week and 3-week experimental group, the amount of the latter group being less than that of the former group; however, no expression of specific fused protein was found in the 2 control groups. Pathological examination showed that the narrowing of lumen 1-week and 3-week experimental groups were 11.50% and 19.75%, both significantly milder than those of the 1-week and 3-week control groups (33.25% and 52.25% respectively, both f P < 0.05). VB staining showed that the (intima/media (I/M) ratio of the 1-week and 3-week experimental groups were 0.12 and 0.35 respectively, both significantly lower than those of the 2 control groups (0.50 and 1.07 respectively, both P < 0.05). CONCLUSION: Accelerating re-endothelialization and inhibiting thrombosis, the fused gene hVEGF(165)-FH effectively prevents restenosis after angioplasty, On the basis of endothelial repair, construction of fused genes with double even multiple targets may be a novel and potential therapeutic approach for restenosis after percutaneous coronary intervention.