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As a circadian organ, liver functions are regulated by circadian clock. Here, the authors present a comprehensive proteomics landscape of the mouse liver, including transcription factor binding profiles, phosphorylation and ubiquitylation patterns, nuclear and whole proteome, and the transcriptome.

Eukaryotic cells store neutral lipids in cytoplasmic lipid droplets (LDs) enclosed in a monolayer of phospholipids and associated proteins [LD proteins (LDPs)]. Growing evidence has demonstrated that LDPs play important roles in the pathogenesis of liver diseases. However, the composition of liver LDPs and the role of their alterations in hepatosteatosis are not well-understood. In this study, we performed liver proteome and LD sub-proteome profiling to identify enriched proteins in LDs as LDPs, and quantified their changes in a high-fat diet (HFD)-induced fatty liver model. Among 5,000 quantified liver proteins, 101 were enriched by greater than 10-fold in the LD sub-proteome and were classified as LDPs. Differential profiling of LDPs in HFD-induced fatty liver provided a list of candidate LDPs for functional investigation. We tested the function of an upregulated LDP, S100a10, in vivo with adenovirus-mediated gene silencing and found, unexpectedly, that knockdown of S100a10 accelerated progression of HFD-induced liver steatosis. The S100A10 interactome revealed a connection between S100A10 and lipid transporting proteins, suggesting that S100A10 regulates the development and formation of LDs by transporting and trafficking. This study identified LD-enriched sub-proteome in homeostatic as well as HFD-induced fatty livers, providing a rich resource for the LDP research field.

The invasion-metastasis cascade is one of the most important factors relating to poor survival and prognosis of malignant melanoma (MM) patients. Long non-coding RNA lymph node metastasis associated transcript 1 (LNMAT1) is a key regulator in lymph node metastasis of multiple cancer types, but the roles and underlying mechanisms of LNMAT1 in the invasion-metastasis cascade of MM remain unclear. In the present study, we aimed to investigate the expression and function of LNMAT1 in MM. Here, we found that LNMAT1 was upregulated in MM tissues and cells, and its expression levels were further enhanced in MM patients with lymph node metastasis and metastatic MM cells. Using loss-of-function assays, we found that LNMAT1 promoted cell migration and invasion and lung metastasis in MM in vitro and in vivo. Moreover, we found that cell adhesion molecule 1 (CADM1), the established tumor suppressor in MM, was the downstream target of LNMAT1. Mechanistically, LNMAT1 epigenetically suppressed CADM1 expression by recruiting EZH2, the key regulator of trimethylation of histone H3 at lysine 27 (H3K27me3), to the CADM1 promoter, resulting in transcriptional inhibition of CADM1. Lastly, rescue assays demonstrated that LNMAT1 promoted cell migration and invasion of MM by suppressing CADM1 expression. Our findings elucidate a new mechanism for LNMAT1-mediated invasion-metastasis cascade in MM and suggest that LNMAT1 may be a new therapeutic target and prognostic predictor for MM.

IntroductionGenetic polymorphisms are important in explaining the wide interpatient variability that exists in the development of acute kidney injury (AKI) post cardiac surgery. We hypothesised that polymorphisms in 4 candidate genes, namely angiotensin-converting enzyme (ACE), apolipoprotein-E (ApoE), interleukin-6 (IL-6), and tumour necrosis factor-alpha (TNF-α) are associated with AKI.Methods870 patients who underwent cardiac surgery in Singapore were analysed. All patients who fulfilled stage 1 KDIGO criteria and above were considered to have AKI. This was investigated against various demographic, clinical and genetic factors.ResultsIncreased age, history of hypertension, anaemia and renal impairment remained important preoperative risk factors for AKI. Intraoperatively, longer cardiopulmonary bypass (CPB) time and the use of intra-aortic balloon pump (IABP) were shown to be associated with AKI. Among the genetic factors, ACE-D allele was associated with an increased risk of AKI while IL6-572C allele was associated with a decreased risk of AKI.ConclusionACE-D allele was associated with the development of AKI similar to other studies. On the other hand, IL6-572C was shown to have a protective role against the development of AKI, contradictory to studies done in the Caucasian population. This contradictory effect of IL6-572C is a result of a complex interplay between the gene and population specific modulating factors. Our findings further underscored the necessity of taking into account population specific differences when developing prediction models for AKI.

Vascular endothelial cells produce reactive oxygen species (ROS) during the process of energy metabolism in aerobic respiration. A growing body of evidence indicates that excessive ROS is implicated in the pathogenesis of cardiovascular diseases including atherosclerosis. The newly synthesized halophenol, 2,4′,5′-trihydroxyl-5,2′-dibromo diphenylmethanone (TDD), exhibits antioxidative and cytoprotective activities in vitro. In this study, the protective effect of TDD against hydrogen peroxide (H2O2)-induced oxidative injury of EA.hy926 cells was investigated. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-dephenyltetrazolium bromide (MTT) assay, while the effect of TDD on the transcription profile of EA.hy926 cells subjected to H2O2-induced oxidative injury was evaluated by microarray analysis. Several signaling pathways, including apoptosis, were significantly associated with TDD. Flow cytometric analysis was used to evaluate anti-apoptotic effect of TDD. Subsequently, RT-PCR and Western blot were used to detect the expressions of the apoptosis-associated protein, Bcl-2 and Bax. Meanwhile the expression of cleaved caspase-3, an executioner of apoptosis, was also detected by Western blot. The results showed that pretreatment of EA.hy926 cells with TDD prevented the decrease of cell viability induced by H2O2, and attenuated H2O2-induced elevation of Bax and cleaved caspase-3 while increased Bcl-2 expressions. In summary, TDD inhibited H2O2-induced oxidative injury of EA.hy926 cells through negative regulation of apoptosis. These findings suggest that TDD is a potential candidate for therapeutic intervention in oxidative stress-associated cardiovascular diseases.

It has been shown that melatonin may affect bone metabolism. However, it is controversial whether melatonin could promote osteoblast proliferation, and the precise molecular mechanism of melatonin on osteoblast proliferation is still obscure. In this study, the results of the CCK-8 assay showed that melatonin significantly promoted human osteoblastic cell line hFOB 1.19 cell proliferation at 1, 2.5, 5, 10, 25, 50 and 100 µM concentrations for 24 h, but there were no significant differences among the groups. Western blot demonstrated that 10 µM melatonin significantly promoted ERK1/2 phosphorylation. Furthermore, we also detected the phosphorylation of c-Raf, MEK1/2, p90RSK and MSK1, and all of them increased with 10 µM melatonin. U0126 (a selective inhibitor of MEK that disrupts downstream activation of ERK1/2) downregulated the phosphorylation of ERK1/2, p90RSK and MSK1. U0126 also attenuated the proliferation of osteoblasts stimulated by melatonin. In conclusion, this study for the first time indicates that melatonin (10 nM–100 µM) promotes the proliferation of a human osteoblastic cell line hFOB 1.19 through activation of c-Raf, MEK1/2, ERK1/2, p90RSK and MSK1.

When the fatigue cycle is extended from high cycle (105~107) to very high cycle (107~109), the fatigue testing results will be more sensitive to the influential factors such as specimen size, specimen surface roughness and the inclusion size. The influential factors on the very high cycle fatigue testing results are investigated in the present paper. Firstly, the design and control method for ultrasonic fatigue test were introduced for several specimen shapes. The effect of the shape, size and the surface roughness of specimen on the ultrasonic fatigue test results are investigated. Meanwhile, the effect of test frequency and the size of the inclusion on the fatigue test results are also investigated. It is concluded that: 1. the test results of specimen with different shape and size differ with each other, due to the risk volume is different. 2. There is a critical roughness for the specimen, depending on the hardness of tested metallic material. A larger roughness than the critical one will lead to a premature fracture. 3. The frequency effect is obvious for the low strength steel, however, is prone to vanish for the very high strength steel. 4. The very high cycle fatigue will be more sensitive to the inclusion size as the strength increases and the S/N curve character is strongly related to the size of the inclusion.

Oxidative stress plays a critical role in endothelial injury and the pathogenesis of diverse cardiovascular diseases, including atherosclerosis. Isoquercitrin (quercetin-3-glucoside), a flavonoid distributed widely in plants, exhibits many biological activities, including anti-allergic, anti-viral, anti-inflammatory, and anti-oxidative effects. In the present study, the inhibitory effect of isoquercitrin on H2O2-induced apoptosis of EA.hy926 cells was evaluated. MTT assays showed that isoquercitrin significantly inhibited H2O2-induced loss of viability in EA.hy926 cells. Hoechst33342/PI and Annexin V-FITC/PI fluorescent double staining indicated that isoquercitrin inhibited H2O2-induced apoptosis of EA.hy926 cells. Western blotting demonstrated that isoquercitrin prevented H2O2-induced increases in cleaved caspase-9 and cleaved caspase-3 expression, while increasing expression of anti-apoptotic protein Mcl-1. Additionally, isoquercitrin significantly increased the expression of p-Akt and p-GSK3β in a dose-dependent manner in EA.hy926 cells. LY294002, a PI3K/Akt inhibitor, inhibited isoquercitrin-induced GSK3β phosphorylation and increase of Mcl-1 expression, which indicated that regulation of isoquercitrin on Mcl-1 expression was likely related to the modulation of Akt activation. These results demonstrated that the anti-apoptotic effect of isoquercitrin on H2O2-induced EA.hy926 cells was likely associated with the regulation of isoquercitrin on Akt/GSK3β signaling pathway and that isoquercitrin could be used clinically to interfere with the progression of endothelial injury-associated cardiovascular disease.

The 10 MeV accelerator-driven subcritical system (ADS) Injector I test stand at Institute of High Energy Physics (IHEP) is a testing facility dedicated to demonstrate one of the two injector design schemes [Injector Scheme-I, which works at 325 MHz], for the ADS project in China. The injector is composed of two parts, the linac part and the beam dump line. The former is designed on the basis of 325 MHz four-vane type copper structure radio frequency quadrupole and superconducting (SC) spoke cavities with β=0.12. The latter is designed to transport the beam coming out of the SC section of the linac to the beam dump, where the beam transverse profile is fairly enlarged and unformed to simplify the beam target design. The SC section consists of two cryomodules with 14 β=0.12 Spoke cavities, 14 solenoid and 14 BPMs in total. The first challenge in the physics design comes from the necessary space required for the cryomodule separation where the periodical lattice is destroyed at a relatively lower energy of ∼5 MeV. Another challenge is the beam dump line design, as it will be the first beam dump line being built by using a step field magnet for the transverse beam expansion and uniformity in the world. This paper gives an overview of the physics design study together with the design principles and machine construction considerations. The results of an optimized design, fabrication status and end to end simulations including machine errors are presented.

The new synthetic anti-inflammatory agent, 4-(5'-dimethylamino)-naphthalenesulfonyl-2(3H)-benzoxazolone (W3D), has demonstrated definite biological activity both in vitro and in vivo. In this study, we investigated the antinociceptive effects of W3D on various animal pain models. Our results showed that W3D, at doses of 12.5 and 25 mg/kg, significantly reduced acetic acidinduced mice writhing by 65.9% and inhibited the pain response in the second phase of formalin-induced pain from 167.75 ± 6.76 s to 97.25 ±9.63 s. Moreover, W3D prolonged the latent period on the tail-immersion test, which was stronger than the positive drug, aspirin (50 mg/kg). However, W3D did not show analgesic activity on the hot plate test, indicating no central pain response. Furthermore, the antinociceptive effects of W3D were not antagonized by naloxone, EGTA, CaCl2, and reserpine. However, LArginine was able to reverse the antinociceptive effect, indicating that W3D's antinociceptive effect does not depend on opioid receptors, Ca2+ concentration, or monoamine content. Instead, it is closely associated with nitric oxide (NO) content. In addition, W3D was found to alleviate nocifensive behavior and improve brain histopathology by inhibiting NO, inducible nitric oxide synthase (iNOS), and calcitonin gene-related peptide in nitroglycerin-induced migraine mice, with more than 50% inhibition ratio at only 12.5 mg/kg orally. However, noradrenaline, dopamine, and 5-hydroxytryptamine levels were not significantly altered after treatment with W3D. In conclusion, the presented data suggest that W3D can be considered a new non-steroidal anti-inflammatory and antinociceptive agent for the treatment of peripheral pain and migraine mediated by NO and iNOS. [ABSTRACT FROM AUTHOR]