HMGCS2-Induced Autophagic Degradation of Tau Involves Ketone Body and ANKRD24.

BACKGROUND: Accumulation of hyperphosphorylated Tau (pTau) contributes to the formation of neurofibrillary tangles in Alzheimer's disease (AD), and targeting Tau/pTau metabolism has emerged as a therapeutic approach. We have previously reported that mitochondrial 3-hydroxy-3-methylglutaryl-COA synthase 2 (HMGCS2) is involved in AD by promoting autophagic clearance of amyloid-ß protein precursor via ketone body-associated mechanism, whether HMGCS2 may also regulate Tau metabolism remains elusive. OBJECTIVE: The present study was to investigate the role of HMGCS2 in Tau/p degradation. METHODS: The protein levels of Tau and pTau including pT217 and pT181, as well as autophagic markers LAMP1 and LC3-II were assessed by western blotting. The differentially regulated genes by HMGCS2 were analyzed by RNA sequencing. Autophagosomes were assessed by transmission electron microscopy. RESULTS: HMGCS2 significantly decreased Tau/pTau levels, which was paralleled by enhanced formation of autophagic vacuoles and prevented by autophagic regulators chloroquine, bafilomycin A1, 3-methyladenine, and rapamycin. Moreover, HMGCS2-induced alterations of LAMP1/LC3-II and Tau/pTau levels were mimicked by ketone body acetoacetate or ß-hydroxybutyrate. Further RNA-sequencing identified ankyrin repeat domain 24 (ANKRD24) as a target gene of HMGCS2, and silencing of ANKRD24 reduced LAMP1/LC3-II levels, which was accompanied by the altered formation of autophagic vacuoles, and diminished the effect of HMGCS2 on Tau/pTau. CONCLUSION: HMGCS2 promoted autophagic clearance of Tau/pTau, in which ketone body and ANKRD24 played an important role.

miR-3651 Participates in the Growth Cycle of Hepatocellular Carcinoma Cells and Promotes the Malignant Metastasis via the PI3K/AKT/mTOR Signalling Pathway.

Objective: Hepatocellular carcinoma (HCC) shows a growing incidence over the past few years, and clinical efforts are made to search for more effective novel diagnosis and therapy regimen for it to improve its outcome. This study probed into the association of miR-3651 with the PI3K/AKT/mTOR pathway to offer a more detailed reference to the follow-up exploration of novel diagnosis and therapy methods of HCC. Methods: Totally, 83 patients with HCC treated in our hospital between Apr. 2017 and Aug. 2018, 100 patients with simple liver cirrhosis (LC), and 94 normal persons over the same time span were enrolled, and serum miR-3651 in them was quantified to understand the predictive and prognostic significance of miR-3651 for HCC. In addition, with purchased human HCC cell strains (HepG2), the impacts of miR-3651 on the invasion as well as proliferation of HepG2 were determined using the MTT and Transwell assays, and the PI3K/AKT/mTOR pathway and autophagy-associated proteins in HepG2 were quantified via WT. Results: Serum miR-3651 was found to be higher in HCC patients than in LC patients and normal persons, and it presented a sensitivity and specificity of 57.14% and 94.00%, respectively, in forecasting the occurrence of HCC in LC patients. The decrease of miR-3651 in HCC patients after therapy was strongly bound up with patients' prognosis, and its increase implied an increased risk of death. In in vitro assays, HepG2 presented higher miR-3651 expression than HL-7702, and upregulated miR-3651 intensified the invasion and proliferation of HepG2, while silencing miR-3651 gave rise to opposite results. Additionally, the PI3K/Akt/mTOR pathway in HepG2 presented an obvious activation state, and its activation was further intensified after increase of miR-3651, while its activation was suppressed after silence of miR-3651. Moreover, HepG2 presented notably downregulated autophagy-associated proteins, and the increase of miR-3651 further suppressed the autophagy process, but with the intervention of BEZ235, the impacts of miR-3651 were completely reversed. Conclusion: miR-3651 intensifies the growth and invasion of HCC cells through activating the PI3K/AKT/mTOR signalling pathway, which is probably a breakthrough in the future diagnosis and therapy of HCC.

The superior lubricating performance and unique mechanism of oil-soluble protic ionic liquids with short alkyl chains.

HYPOTHESIS: Ionic liquids (ILs), as lubricant additives, can greatly improve the lubricating behavior of the frictional interfaces. However, it is urgent to explore ILs with good oil solubility in nonpolar oils, and it is necessary to further study and verify the lubrication mechanism of ILs from the perspective of alkyl chain length. EXPERIMENTS: Five protic ILs (PILs) with varying alkyl chain lengths were synthesized by proton transfer method. As additives in PAO oil, their tribological properties were investigated on SRV-V tester. Through molecular dynamics simulation, the adsorption behavior of PILs at the frictional interface was illustrated. FINDINGS: The tribological properties of base oil could be significantly improved by adding PIL additives, but interestingly, PILs with short-chain anions showed better lubricating performance, which contradicted most of the early findings. Further analyses revealed that PILs achieved effective lubrication by the tribochemical interaction between anions and frictional interface, and the formation of cationic protective layer. However, PILs with shorter-chain anions form a denser protective layer that can better support the interfacial anions to participate in tribochemical reactions and thus abnormally exhibit superior lubricating performance than those with longer-chain anions.

The relationship between solid-liquid interface interaction and gelling capacity of h-BN 2D material: a rheological study.

Diurea modified h-BN nanosheet is a novel kind of 2D gelator that could gel the lubricating oils under the stimulus of ultrasound. Morphological analyzations in previous study confirmed that the ultrasound induced layer-by-layer (LBL) structure of BN gelator is critical for the gelation. However, the elastic response in LBL structure, which is crucial for the formation of a stable gel system, has not been explicitly illustrated yet. The challenge is that the LBL gelator structure is based on 2D material and thus lacks vertical linkage between gelator layers, which is significantly different from the traditional gel systems that generally possess highly crosslinked gelator network. In this work, by investigating the viscoelastic behavior of the BN-based gel via rheometer, it is found the solid-liquid interface interaction, which is regulated by the diurea molecular structure in the BN gelator, is the key factor for triggering the stable elastic response in the LBL structure, and the elasticity mainly originates from the interface interaction induced bending deformation of h-BN 2D material. The findings further elucidate the gelling mechanism of BN gelators and enlighten the structure design of ultrasound-responsive gelator based on 2D materials.

Sulfuretin exerts diversified functions in the processing of amyloid precursor protein.

Sulfuretin is a flavonoid that protects cell from damage induced by reactive oxygen species and inflammation. In this study, we investigated the role of sulfuretin in the processing of amyloid precursor protein (APP), in association with the two catalytic enzymes the α-secretase a disintegrin and metalloproteinase (ADAM10), and the beta-site APP cleaving enzyme 1 (BACE1) that play important roles in the generation of ß amyloid protein (Aß) in Alzheimer's disease (AD). We found that sulfuretin increased the levels of the immature but not the mature form of ADAM10 protein. The enhanced ADAM10 transcription by sulfuretin was mediated by the nucleotides -444 to -300 in the promoter region, and was attenuated by silencing or mutation of transcription factor retinoid X receptor (RXR) and by GW6471, a specific inhibitor of peroxisome proliferator-activated receptor α (PPAR-α). We further found that sulfuretin preferentially increased protein levels of the immature form of APP (im-APP) but significantly reduced those of BACE1, sAPPß and ß-CTF, whereas Aß1-42 levels were slightly increased. Finally, the effect of sulfuretin on BACE1 and im-APP was selectively attenuated by the translation inhibitor cycloheximide and by lysosomal inhibitor chloroquine, respectively. Taken together, (1) RXR/PPAR-α signaling was involved in sulfuretin-mediated ADAM10 transcription. (2) Alteration of Aß protein level by sulfuretin was not consistent with that of ADAM10 and BACE1 protein levels, but was consistent with the elevated level of im-APP protein, suggesting that im-APP, an isoform mainly localized to trans-Golgi network, plays an important role in Aß generation.

TUFM is involved in Alzheimer's disease-like pathologies that are associated with ROS.

Mitochondrial Tu translation elongation factor (TUFM or EF-Tu) is part of the mitochondrial translation machinery. It is reported that TUFM expression is reduced in the brain of Alzheimer's disease (AD), suggesting that TUFM might play a role in the pathophysiology. In this study, we found that TUFM protein level was decreased in the hippocampus and cortex especially in the aged APP/PS1 mice, an animal model of AD. In HEK cells that stably express full-length human amyloid-ß precursor protein (HEK-APP), TUFM knockdown or overexpression increased or reduced the protein levels of ß-amyloid protein (Aß) and ß-amyloid converting enzyme 1 (BACE1), respectively. TUFM-mediated reduction of BACE1 was attenuated by translation inhibitor cycloheximide (CHX) or α-[2-[4-(3,4-Dichlorophenyl)-2-thiazolyl]hydrazinylidene]-2-nitro-benzenepropanoic acid (4EGI1), and in cells overexpressing BACE1 constructs deleting the 5' untranslated region (5'UTR). TUFM silencing increased the half-life of BACE1 mRNA, suggesting that RNA stability was affected by TUFM. In support, transcription inhibitor Actinomycin D (ActD) and silencing of nuclear factor κB (NFκB) failed to abolish TUFM-mediated regulation of BACE1 protein and mRNA. We further found that the mitochondria-targeted antioxidant TEMPO diminished the effects of TUFM on BACE1, suggesting that reactive oxygen species (ROS) played an important role. Indeed, cellular ROS levels were affected by TUFM knockdown or overexpression, and TUFM-mediated regulation of apoptosis and Tau phosphorylation at selective sites was attenuated by TEMPO. Collectively, TUFM protein levels were decreased in APP/PS1 mice. TUFM is involved in AD pathology by regulating BACE1 translation, apoptosis, and Tau phosphorylation, in which ROS plays an important role.

Mitochondrial TXN2 attenuates amyloidogenesis via selective inhibition of BACE1 expression.

Thioredoxin-2 (TXN2) is a mitochondrial protein and represents one of the intrinsic antioxidant enzymes. It has long been recognized that mitochondrial dysfunction and oxidative stress contribute to the pathogenesis of Alzheimer's disease (AD). We hypothesized that mitochondrial TXN2 might play a role in AD-like pathology. In this study, we found that in SH-SY5Y and HEK cells stably express full-length human amyloid-ß precursor protein (HEK-APP), TXN2 silencing or over-expression selectively increased or decreased the transcription of beta-site amyloid precursor protein cleaving enzyme 1 (BACE1), respectively, without altering the protein levels of others enzymes involved in the catalytic processing of APP. As a result, ß-amyloid protein (Aß) levels were significantly decreased by TXN2. In addition, in cells treated with 3-nitropropionic acid (3-NP) that is known to increase reactive oxygen species (ROS) and promote mitochondrial dysfunction, TXN2 silencing resulted in further enhancement of BACE1 protein levels, suggesting a role of TXN2 in ROS removal. The downstream signaling might involve NFκB, as TXN2 reduced the phosphorylation of p65 and IκBα; and p65 knockdown significantly attenuated TXN2-mediated regulation of BACE1. Concomitantly, the levels of cellular ROS, apoptosis-related proteins and cell viability were altered by TXN2 silencing or over-expression. In APPswe/PS1E9 mice, an animal model of AD, the cortical and hippocampal TXN2 protein levels were decreased at 12 months but not at 6 months, suggesting an age-dependent decline. Collectively, TXN2 regulated BACE1 expression and amyloidogenesis via cellular ROS and NFκB signaling. TXN2 might serve as a potential target especially for early intervention of AD.

Construction of a continuously layered structure of h-BN nanosheets in the liquid phase via sonication-induced gelation to achieve low friction and wear.

Herein, to endow h-BN nanosheets with gelling ability, a diurea compound was decorated on the h-BN nanosheets via designed adsorption and in situ reaction processes. The prepared h-BN-based gelator, BTO, exhibited excellent dispersibility in non-polar liquid media, and the gelation of BTO dispersions could be readily triggered by ultrasonic treatments. The sol-gel transformation of the system was found to be highly reversible by stirring and sonication. Based on the investigation on the self-assembly behavior of BTO nanosheets in the liquid phase, it was proposed that a continuous and layered structure formed by BTO during sonication was the key factor for the gelling properties of these nanosheets. The viscoelasticity of the sonication-induced gel was studied using a rheometer. Tribological evaluations show that the prepared h-BN nanogel exhibits outstanding lubricating performances, and more importantly, it has been proved that the gel state of the h-BN nanosheets provides superior and more reliable lubricating performances than the corresponding dispersion state under certain conditions; this can be ascribed to the formation of a continuous and uniform structure of modified h-BN nanosheets during gelation. Thus, this study not only clarifies the key role of the assembly structure in the tribological performances of 2D nanomaterials, but also demonstrates the potential of gelation in improving the macroscopic friction reduction and wear resistance of 2D nanomaterials.

Novel Approach to the Fabrication of an Alumina-MoS2 Self-Lubricating Composite via the In Situ Synthesis of Nanosized MoS2.

The fabrication of a self-lubricating ceramic composite with MoS2 as the solid lubricant is extremely difficult given the high temperature sensitivity of MoS2. In this study, a hydrothermal method was utilized for the in situ synthesis of nanosized-MoS2 in Al2O3 ceramic to fabricate an Al2O3-MoS2 self-lubricating composite. The composite exhibited excellent self-lubricating properties with low friction coefficient and wear rate in a high-vacuum environment because of the efficient formation of self-lubricating films from the finely structured MoS2 in the Al2O3 matrix. The results of this study establish a new approach to the fabrication of self-lubricating ceramic composites with temperature-sensitive solid lubricants.

HMGCS2 promotes autophagic degradation of the amyloid-ß precursor protein through ketone body-mediated mechanisms.

HMGCS2 (mitochondrial 3-hydroxy-3-methylglutaryl-COA synthase 2) is a control enzyme in ketogenesis. The mitochondrial localization and interaction with APP (ß-amyloid precursor protein) suggest that HMGCS2 may play a role in the pathophysiology of AD (Alzheimer's disease). Here we report that overexpression of HMGCS2 decreased levels of APP and related CTFs (carboxy-terminal fragments), which was largely prevented by an autophagic inhibitor chloroquine. In addition, HMGCS2 enhancement of autophagic marker LC3II was diminished by rapamycin, an inhibitor of mechanistic target of rapamycin. Moreover, deprivation of EBSS (Earle's Balanced Salt Solution) significantly augmented the effect of HMGCS2 on LC3II, while acetoacetate reversed the reduction of LC3II, APP and CTFs which was induced by HMGCS2 knockdown. In the presence of acetoacetate, rapamycin failed to induce further increase of LC3II, which mimicked the effect of HMGCS2 overexpression. Finally, HMGCS2 enhanced the antioxidant response. Collectively, HMGCS2 shares with ketone bodies common features in autophagic clearance of APP and CTFs, suggesting that ketone bodies play an important role in HMGCS2 regulation of the autophagy.

Improved Tribological Performance of Amorphous Carbon (a-C) Coating by ZrO2 Nanoparticles.

Nanomaterials, such as Graphene, h-BN nanoparticles and MoS2 nanotubes, have shown their ability in improving the tribological performance of amorphous carbon (a-C) coatings. In the current study, the effectiveness of ZrO2 nanoparticles (ZrO2-NPs) in lubricating the self-mated nonhydrogenated a-C contacts was investigated in boundary lubrication regime. The results showed that 13% less friction and 50% less wear compared to the base oil were achieved by employing ZrO2-NPs in the base oil in self-mated a-C contacts. Via analyzing the ZrO2-NPs and the worn a-C surface after tests, it was found that the improved lubrication by ZrO2-NPs was based on "polishing effects", which is a new phenomenon observed between a-C and nanoparticles. Under the "polishing effect", micro-plateaus with extremely smooth surface and uniform height were produced on the analyzed a-C surface. The resulting topography of the a-C coating is suitable for ZrO2-NPs to act as nano-bearings between rubbing surfaces. Especially, the ZrO2-NPs exhibited excellent mechanical and chemical stability, even under the severe service condition, suggesting that the combination of nonhydrogenated a-C coating with ZrO2-NPs is an effective, long lasting and environment-friendly lubrication solution.

Long noncoding RNA GAS5 suppresses the migration and invasion of hepatocellular carcinoma cells via miR-21.

Long noncoding RNAs (lncRNAs) are aberrantly expressed in various cancers. Although lncRNA GAS5 (growth arrest-specific transcript 5) has been characterized as a tumor suppressor in some kinds of cancer, its role and function in hepatocellular carcinoma (HCC) remain unknown. The present report demonstrates that there are lower levels of GAS5, PDCD4, and PTEN and higher levels of microRNA-21 (miR-21) in HCC tissues than in adjacent normal tissues. Moreover, the levels of GAS5 and miR-21 were correlated with the clinicopathological characteristics of HCC. HCC patients with higher levels of GAS5 or with the lower levels of miR-21 have longer survival times. There are lower levels of GAS5 and higher levels of miR-21 in HCC cell lines (Be7402, SMMC-7721, and HCCLM3) than in normal liver L-02 cells, and the levels correlate with the aggression of the HCC cell lines. Knockdown of GAS5 upregulates miR-21 levels in Bel-7402 cells (weakly aggressive); in contrast, there are opposite changes in HCCLM3 cells (highly aggressive). Moreover, GAS5 that upregulated or downregulated the expression of PDCD4 and PTEN was reversed by inhibiting or overexpressing miR-21 level in Bel-7402 and HCCLM3 cells. Then, overexpression of GAS5 suppresses the migration and invasion of HCC cells and high expression of miR-21 largely eliminates GAS5-mediated suppression of HCC cell migration and invasion. Thus, GAS5 acts as a tumor suppressor in HCCs through negative regulation of miR-21 and its targets and proteins about migration and invasion in cancer cells, which may be a target for treating HCC.

Low-frequency repetitive transcranial magnetic simulation prevents chronic epileptic seizure.

Although low-frequency repetitive transcranial magnetic simulation can potentially treat epilepsy, its underlying mechanism remains unclear. This study investigated the influence of low-frequency re-petitive transcranial magnetic simulation on changes in several nonlinear dynamic electroence-phalographic parameters in rats with chronic epilepsy and explored the mechanism underlying petitive transcranial magnetic simulation-induced antiepileptic effects. An epilepsy model was es-tablished using lithium-pilocarpine intraperitoneal injection into adult Sprague-Dawley rats, which were then treated with repetitive transcranial magnetic simulation for 7 consecutive days. Nonlinear electroencephalographic parameters were obtained from the rats at 7, 14, and 28 days post-stimulation. Results showed significantly lower mean correlation-dimension and Kolmogo-rov-entropy values for stimulated rats than for non-stimulated rats. At 28 days, the complexity and point-wise correlation dimensional values were lower in stimulated rats. Low-frequency repetitive transcranial magnetic simulation has suppressive effects on electrical activity in epileptic rats, thus explaining its effectiveness in treating epilepsy.

Fabrication of a nanocrystalline Ni-Co/CoO functionally graded layer with excellent electrochemical corrosion and tribological performance.

Nanocrystalline (NC) Ni-Co/CoO functionally graded materials with excellent lubricating, high anti-corrosion and anti-wear performance were fabricated by electrodeposition and subsequent cyclic thermal oxidation and quenching. Transmission electron microscopy and energy dispersive x-ray spectroscopy investigations show that bulk Ni-Co gradient deposits with an average grain size in the range of 13-40 nm demonstrated a graded structure transition from face-centred cubic to hexagonal close packed and graded composition changes from Ni-rich to Co-rich regions with the increase in deposit thickness. X-ray diffraction and x-ray photoelectron spectroscopy analysis indicated the surface layer of NC Ni-Co graded materials to be mainly composed of dense and ultrafine CoO with a (111) preferred orientation. The NC Ni-Co/CoO functionally graded materials exhibited significantly enhanced corrosion resistance in both NaOH and NaCl solutions and remarkably improved wear resistance and dry self-lubricating performance when compared with the NC Ni and Ni-Co graded deposits under dry sliding wear conditions. The higher corrosion and tribological performance of NC Ni-Co/CoO graded materials can be attributed to the graded microstructure within the deposits, the anti-corrosion barrier of a dense oxide layer and the solid lubrication effect of CoO-rich tribo-surface films.