Mining cholesterol genes from thousands of mouse livers identifies aldolase C as a regulator of cholesterol biosynthesis.

The availability of genome-wide transcriptomic and proteomic datasets is ever-increasing and often not used beyond initial publication. Here, we applied module-based coexpression network analysis to a comprehensive catalog of 35 mouse genome-wide liver expression datasets (encompassing more than 3800 mice) with the goal of identifying and validating unknown genes involved in cholesterol metabolism. From these 35 datasets, we identified a conserved module of genes enriched with cholesterol biosynthetic genes. Using a systematic approach across the 35 datasets, we identified three genes (Rdh11, Echdc1, and Aldoc) with no known role in cholesterol metabolism. We then performed functional validation studies and show that each gene is capable of regulating cholesterol metabolism. For the glycolytic gene, Aldoc, we demonstrate that it contributes to de novo cholesterol biosynthesis and regulates cholesterol and triglyceride levels in mice. As Aldoc is located within a genome-wide significant genome-wide association studies locus for human plasma cholesterol levels, our studies establish Aldoc as a causal gene within this locus. Through our work, we develop a framework for leveraging mouse genome-wide liver datasets for identifying and validating genes involved in cholesterol metabolism.

Control of NAD+ homeostasis by autophagic flux modulates mitochondrial and cardiac function.

Impaired autophagy is known to cause mitochondrial dysfunction and heart failure, in part due to altered mitophagy and protein quality control. However, whether additional mechanisms are involved in the development of mitochondrial dysfunction and heart failure in the setting of deficient autophagic flux remains poorly explored. Here, we show that impaired autophagic flux reduces nicotinamide adenine dinucleotide (NAD+) availability in cardiomyocytes. NAD+ deficiency upon autophagic impairment is attributable to the induction of nicotinamide N-methyltransferase (NNMT), which methylates the NAD+ precursor nicotinamide (NAM) to generate N-methyl-nicotinamide (MeNAM). The administration of nicotinamide mononucleotide (NMN) or inhibition of NNMT activity in autophagy-deficient hearts and cardiomyocytes restores NAD+ levels and ameliorates cardiac and mitochondrial dysfunction. Mechanistically, autophagic inhibition causes the accumulation of SQSTM1, which activates NF-κB signaling and promotes NNMT transcription. In summary, we describe a novel mechanism illustrating how autophagic flux maintains mitochondrial and cardiac function by mediating SQSTM1-NF-κB-NNMT signaling and controlling the cellular levels of NAD+.

Coil Positioning for Wireless Power Transfer System of Automatic Guided Vehicle Based on Magnetic Sensing.

As an auxiliary function of the wireless power transfer (WPT) system, coil positioning can solve the power and efficiency degradation during power transmission caused by misalignment of the magnetic coupler. In this paper, a Hall sensor array is used to measure the change of magnetic flux density. By comparing the multisensor data fusion results with the preset data obtained from the coil alignment, the real-time accurate positioning of the receiving coil can be realized. Firstly, the positioning model of the receiving coil is built and the variation of magnetic flux density with the coil misalignment is analyzed. Secondly, the arrangement of the Planar 8-direction symmetric sensor array and the positioning algorithm based on data fusion of magnetic flux density variations are proposed. In order to avoid coil positioning misalignment caused by the unstable magnetic field distribution which is actually affected by the change of mutual inductance during automatic guided vehicle (AGV) alignment, the constant current strategy of primary and secondary sides is proposed. Finally, the coil positioning experimental platform is built. The experimental results show that the coil positioning method proposed in this paper has high accuracy, and the positioning error is within 4 cm.

MicroRNA-4651 represses hepatocellular carcinoma cell growth and facilitates apoptosis via targeting FOXP4.

MicroRNAs (miRNAs) belong to the subgroup of small noncoding RNAs, which typically serve as important gene regulators to participate in different biological events, such as tumor cell growth and apoptosis. Recent studies indicated microRNA-4651 (miR-4651) was involved in hepatocellular carcinoma (HCC) progression. The certain role of miRNA-4651 during the progression of HCC, however, remains unclear. Herein, we investigated the mRNA expression level of miR-4651 in HCC tissues and HCC cell lines and found miR-4651 was noticeably down-regulated compared with the normal liver tissues and QSG-7701 cell line, respectively. Then, miR-4561 overexpression obviously repressed the proliferation and promoted apoptosis in two HCC cell lines. Interestingly, we further identified that miR-4561 could directly interact with FOXP4 in HCC cells by using bio-informatic method and report assay. Moreover, forced expression of FOXP4 showed an opposite effect compared with miR-4561 in HCC cell lines. Hence, our findings strongly indicated that miR-4561 regulated the HCC cell growth and apoptosis mainly through targeting the FOXP4 genes. Clinically, the miR-4561/FOXP4 axis might be a potential target for therapeutic application of HCC patient treatment.

Integrating Mouse and Human Genetic Data to Move beyond GWAS and Identify Causal Genes in Cholesterol Metabolism.

Identifying the causal gene(s) that connects genetic variation to a phenotype is a challenging problem in genome-wide association studies (GWASs). Here, we develop a systematic approach that integrates mouse liver co-expression networks with human lipid GWAS data to identify regulators of cholesterol and lipid metabolism. Through our approach, we identified 48 genes showing replication in mice and associated with plasma lipid traits in humans and six genes on the X chromosome. Among these 54 genes, 25 have no previously identified role in lipid metabolism. Based on functional studies and integration with additional human lipid GWAS datasets, we pinpoint Sestrin1 as a causal gene associated with plasma cholesterol levels in humans. Our validation studies demonstrate that Sestrin1 influences plasma cholesterol in multiple mouse models and regulates cholesterol biosynthesis. Our results highlight the power of combining mouse and human datasets for prioritization of human lipid GWAS loci and discovery of lipid genes.

Flow Diverter-Assisted Coil Embolization of Blood Blister-Like Aneurysm Using Semi-deploying Technique.

Despite many therapeutic methods were utilized to treat blood blister-like aneurysms (BBAs), the optimal treatment approach has not yet been defined. This study presents the single center experience with BBAs treated with flow diverter-assisted coiling using semi-deploying technique, and discusses the efficacy and safety of the method. The patients with subarachnoid hemorrhages (SAH) due to BBAs and treated with Pipeline Flex Embolization Device (PED) between November 2015 and February 2019 in our hospital were retrospectively reviewed. Patient demographic data, timing of treatment, angiographic details, treatment techniques, clinical outcomes and follow-up results were recorded. Ten cases (6 women and 4 men) were enrolled. The mean age of patients was 50.7 years (range 40-61 years). The aneurysm size ranged from 2 × 1.7 mm to 4.5 × 3.8 mm. Seven patients were treated with PED assisted coil embolization using semi-deploying technique, and all of the aneurysms were totally obliterated at the follow up. One patient treated with PED assisted coil embolization suffered from parenchymal hemorrhage 3 days after the treatment, and another one patient also treated with PED and coil died of severe vasospasm 10 days after the treatment. There was no reruptured cases during the follow-up. Here we showed that PED assisted coil embolization using semi-deploying technique could be a technically safe and effective treatment for BBAs.

SNHG29 regulates miR-223-3p/CTNND1 axis to promote glioblastoma progression via Wnt/ß-catenin signaling pathway.

BACKGROUND: Glioblastoma has been seen as the most common malignancy of brain tumor. Emerging reports has claimed that SNHG29 (LRRC75A-AS1) was involved in several biological processes via modulation of signaling pathway, and served as an malignant facilitatorin osteosarcoma. However, the specific role of SNHG29 in glioblastoma remains unknown. METHODS: RT-qPCR and microarray were operated to measure genes expression. Western blot was performed to examine protein expression. CCK-8 and colony formation assays were used to evaluate cell proliferation. Cell migration was tested by transwell assay. Nuclear-cytoplasmic fractionation was conducted to locate SNHG29. The binding capacity of miR-223-3p to SNHG29 or CTNND1 3'UTR was verified by RIP and luciferase reporter assay. RESULTS: SNHG29 presented high expression in glioblastoma to boost cell proliferation, migration and EMT process. In addition, miR-223-3p was validated to bind with SNHG29 after prediction and screening. Furthermore, miR-223-3p was proved to be a negative regulator for its target CTNND1. Then, the inhibition on cell proliferation, migration and EMT process resulted from SNHG29 knockdown was recovered by CTNND1 overexpression. At last, the inhibitive impacts on cell proliferation, migration and EMT process of CTNND1 deficiency was abrogated by LiCl. CONCLUSIONS: In conclusion, SNHG29 regulates miR-223-3p/CTNND1 axis to promote glioblastoma progression via Wnt/ß-catenin signaling pathway, offering a potential therapeutic point for glioblastoma patients.

Protective Effects of Nicorandil on Cardiac Function and Left Ventricular Remodeling in a Rat Model of Ischemic Heart Failure.

Nicorandil, the first clinically applied ATP-sensitive K+ channel (K+ATP) opener with nitrate property, has demonstrated cardioprotective effects in patients with multiples of heart diseases. However, it is unknown whether nicorandil has effects on left ventricular (LV) remodeling in rats with ischemic heart failure and the potential mechanisms remain unclear. In this study, we investigated the effects of nicorandil on cardiac function, LV remodeling, and Bax expression in myocardium of LV in rats with ischemic heart failure. We found that nicorandil could improve not only the general condition, but also the cardiac function in rats with ischemic heart failure. The data also demonstrated that nicorandil reduced the hypertrophy and fibrosis of LV in rats with ischemic heart failure. Furthermore, nicorandil suppressed the protein level of Bax expression in LV myocardium. Taken together, these results suggest that nicorandil exerts its cardioprotective effect and improves LV remodeling in rats with ischemic heart failure. The mechanism might be relative to the inhibitory effect of nicorandil on the protein level of Bax expression in LV myocardium.

Nicotinamide riboside is uniquely and orally bioavailable in mice and humans.

Nicotinamide riboside (NR) is in wide use as an NAD+ precursor vitamin. Here we determine the time and dose-dependent effects of NR on blood NAD+ metabolism in humans. We report that human blood NAD+ can rise as much as 2.7-fold with a single oral dose of NR in a pilot study of one individual, and that oral NR elevates mouse hepatic NAD+ with distinct and superior pharmacokinetics to those of nicotinic acid and nicotinamide. We further show that single doses of 100, 300 and 1,000 mg of NR produce dose-dependent increases in the blood NAD+ metabolome in the first clinical trial of NR pharmacokinetics in humans. We also report that nicotinic acid adenine dinucleotide (NAAD), which was not thought to be en route for the conversion of NR to NAD+, is formed from NR and discover that the rise in NAAD is a highly sensitive biomarker of effective NAD+ repletion.

Adipocyte iron regulates leptin and food intake.

Dietary iron supplementation is associated with increased appetite. Here, we investigated the effect of iron on the hormone leptin, which regulates food intake and energy homeostasis. Serum ferritin was negatively associated with serum leptin in a cohort of patients with metabolic syndrome. Moreover, the same inverse correlation was observed in mice fed a high-iron diet. Adipocyte-specific loss of the iron exporter ferroportin resulted in iron loading and decreased leptin, while decreased levels of hepcidin in a murine hereditary hemochromatosis (HH) model increased adipocyte ferroportin expression, decreased adipocyte iron, and increased leptin. Treatment of 3T3-L1 adipocytes with iron decreased leptin mRNA in a dose-dependent manner. We found that iron negatively regulates leptin transcription via cAMP-responsive element binding protein activation (CREB activation) and identified 2 potential CREB-binding sites in the mouse leptin promoter region. Mutation of both sites completely blocked the effect of iron on promoter activity. ChIP analysis revealed that binding of phosphorylated CREB is enriched at these two sites in iron-treated 3T3-L1 adipocytes compared with untreated cells. Consistent with the changes in leptin, dietary iron content was also directly related to food intake, independently of weight. These findings indicate that levels of dietary iron play an important role in regulation of appetite and metabolism through CREB-dependent modulation of leptin expression.

Preparation and characterization of intravenously injectable curcumin nanosuspension.

The interest in nanosuspensions by the pharmaceutical industry is increasing given several nanosuspension products currently on the market for poorly soluble drugs. In this study, a novel dosage form for curcumin (CUR), CUR nanosuspension (CUR-NS), was successfully prepared by high pressure homogenization to improve CUR's cytotoxicity, as well as improve its application via intravenous injection. Characterization of the CUR-NS was evaluated by morphology, size, zeta potential, solubility, dissolution rate, and crystal state of drug. The nanoparticles for CUR-NS presented a sphere-like shape under transmission electron microscopy with an average diameter of 250.6 nm and the zeta potential of CUR-NS was -27.92 mV. Solubility and dissolution rate of CUR in the form of CUR-NS were significantly increased due to the small particle size and the crystalline state of CUR was preserved to increase its stability against degradation. Superior cytotoxicity in Hela and MCF-7 cells was obtained for CUR-NS compared with CUR solution. The safety evaluation showed that, compared with the CUR solution, CUR-NS provided less local irritation and phlebitis risks, lower rate of erythrocyte hemolysis. These findings suggest that CUR-NS may represent a promising new drug formulation for intravenous administration in the treatment of certain cancers.

A rat model of Shuang Huang Lian injection-induced anaphylaxis.

BACKGROUND: ShuangHuangLian Injection (SHLI) has induced many serious anaphylactic diseases, becoming a threat to the public health. However, study of the mechanism of the reaction and therapeutic approaches to it have been hindered by the lack of suitable animal models. OBJECTIVE: We sought to develop a rat model that could mimic the characteristics of SHLI-induced anaphylaxis and the immunological changes that occur in clinical use. METHODS: Brown Norway (BN) rats were sensitized twice at an interval of seven days with three doses of SHLI and challenged 14 days after the last administration. Different parameters, including the symptoms, the histamine levels and the pathological changes were analyzed. Antibody and cytokine levels were determined to explore the mechanisms involved. RESULTS: Total and SHLI-specific IgE levels were significantly increased after SHLI sensitization. Systemic symptoms, local skin reactions, elevated histamine levels and decreased blood pressures were observed after challenge. Histological examination revealed that slight pathological changes occurred in lungs while no obvious alteration was found in intestine. IL-4 but not IFN-gamma was significantly increased in spleen cells from SHLI-sensitized rats, indicating SHLI-induced anaphylaxis may be Th2-mediated. CONCLUSION: This rat model may provide a useful tool to study the mechanisms in SHLI-induced anaphylaxis and suggest some novel therapeutic approaches for this anaphylaxis.

Effect of chlorogenic acid on mast cell-dependent anaphylactic reaction.

Chlorogenic acid (CGA), a naturally occurring polyphenol compound, has a number of biological activities. However, roles of CGA in the mast cell-dependent anaphylactic reaction have not been fully examined. In the present study, the effect and mechanism of CGA on mast cell-dependent anaphylactic reaction were investigated using in vivo and in vitro models. CGA inhibited compound 48/80-induced systemic anaphylactic shock in mice and skin vascular permeability in rats. CGA also inhibited anti-dinitrophenyl (DNP) IgE-mediated passive cutaneous anaphylaxis (PCA). Moreover, CGA dose-dependently reduced histamine and TNF-alpha release from RBL-2H3 cells activated by anti-DNP IgE. Pretreatment with CGA suppressed IgE-antigen complex induced calcium uptake into RBL-2H3 cells. When CGA was added, the level of intracellular cyclic adenosine monophosphate (cAMP) in RBL-2H3 cells was significantly elevated compared with the untreated cells. Decreased calcium uptake and increased cAMP level might be involved in the inhibitory effect of CGA on mast cell activation. These results suggest a possible therapeutic application of CGA in allergic diseases.

Preparation, characterization, pharmacokinetics, and tissue distribution of curcumin nanosuspension with TPGS as stabilizer.

BACKGROUND: CUR is a promising drug candidate based on its good bioactivity, but use of CUR is potentially restricted because of its poor solubility and bioavailability. AIM: The aim of this study was to prepare an aqueous formulation of curcumin nanosuspension (CUR-NS) to improve its solubility and change its in vivo behavior. METHODS: CUR-NS was prepared by high-pressure homogenization method. Drug state in CUR-NS was evaluated by powder X-ray diffraction. Pharmacokinetics and biodistribution of CUR-NS after intravenous administration in rabbits and mice were studied. RESULTS: The solubility and dissolution of CUR in the form of CUR-NS were significantly higher than those of crude CUR. X-ray crystallography diffraction indicated that the crystalline state of CUR in nanosuspension was preserved. Pharmacokinetics and biodistribution results of CUR-NS after intravenous administration in rabbits and mice showed that CUR-NS presented a markedly different pharmacokinetic property as compared to the CUR solution. AUC(0-infinity) of CUR-NS (700.43 +/- 281.53 microg/mL, min) in plasma was approximately 3.8-fold greater than CUR solution (145.42 +/- 9.29 microg/mL min), and the mean residence time (194.57 +/- 32.18 versus 15.88 +/- 3.56 minutes) was 11.2-fold longer. CONCLUSION: Nanosuspension could serve as a promising intravenous drug-delivery system for curcumin.