CRISPR Screen Identifies the RNA-Binding Protein Eef1a1 as a Key Regulator of Myogenesis.

Skeletal muscle myogenesis hinges on gene regulation, meticulously orchestrated by molecular mechanisms. While the roles of transcription factors and non-coding RNAs in myogenesis are widely known, the contribution of RNA-binding proteins (RBPs) has remained unclear until now. Therefore, to investigate the functions of post-transcriptional regulators in myogenesis and uncover new functional RBPs regulating myogenesis, we employed CRISPR high-throughput RBP-KO (RBP-wide knockout) library screening. Through this approach, we successfully identified Eef1a1 as a novel regulatory factor in myogenesis. Using CRISPR knockout (CRISPRko) and CRISPR interference (CRISPRi) technologies, we successfully established cellular models for both CRISPRko and CRISPRi. Our findings demonstrated that Eef1a1 plays a crucial role in promoting proliferation in C2C12 myoblasts. Through siRNA inhibition and overexpression methods, we further elucidated the involvement of Eef1a1 in promoting proliferation and suppressing differentiation processes. RIP (RNA immunoprecipitation), miRNA pull-down, and Dual-luciferase reporter assays confirmed that miR-133a-3p targets Eef1a1. Co-transfection experiments indicated that miR-133a-3p can rescue the effect of Eef1a1 on C2C12 myoblasts. In summary, our study utilized CRISPR library high-throughput screening to unveil a novel RBP, Eef1a1, involved in regulating myogenesis. Eef1a1 promotes the proliferation of myoblasts while inhibiting the differentiation process. Additionally, it acts as an antagonist to miR-133a-3p, thus modulating the process of myogenesis.

Exploring Multi-Tissue Alternative Splicing and Skeletal Muscle Metabolism Regulation in Obese- and Lean-Type Pigs.

Alternative splicing (AS) is a crucial mechanism in post-transcriptional regulation, contributing significantly to the diversity of the transcriptome and proteome. In this study, we performed a comprehensive AS profile in nine tissues obtained from Duroc (lean-type) and Luchuan (obese-type) pigs. Notably, 94,990 AS events from 14,393 genes were identified. Among these AS events, it was observed that 80% belonged to the skipped exon (SE) type. Functional enrichment analysis showed that genes with more than ten AS events were closely associated with tissue-specific functions. Additionally, the analysis of overlap between differentially alternative splicing genes (DSGs) and differentially expressed genes (DEGs) revealed the highest number of overlapped genes in the heart and skeletal muscle. The novelty of our study is that it identified and validated three genes (PYGM, MAPK11 and CAMK2B) in the glucagon signaling pathway, and their alternative splicing differences were highly significant across two pig breeds. In conclusion, our study offers novel insights into the molecular regulation of diverse tissue physiologies and the phenotypic differences between obese- and lean-type pigs, which are helpful for pig breeding.

Self-Monitoring Theranostic Nanomaterials: Emerging Visual Agents for Real-Time Monitoring of Tumor Treatment Processes.

Self-monitoring in tumor therapy is a concept that allows for real-time monitoring of the location and state of applied nanomaterials. This monitoring relies on dynamic signals, such as wave or magnetic signals, which vary in response to changes in the location and state of nanomaterials. Dynamic changes in nanomaterials can be monitored using dynamic signals, making it possible to determine and control the treatment process. Theranostic nanomaterials, which possess unique physical and chemical properties, have recently been explored as a viable option for self-monitoring. With the help of self-monitoring, theranostic nanomaterials can guide themselves to achieve region-selective treatment with higher controllability and safety. In this review, self-monitoring theranostic nanomaterials will be introduced in three parts according to their roles during therapy: tumor accumulation, tumor therapy, and metabolism. The limitations and future challenges of current self-monitoring theranostic nanomaterials will also be discussed.

Scalable and Flexible Multi-Layer Prismatic Photonic Metamaterial Film for Efficient Daytime Radiative Cooling.

To maintain a comfortable indoor living environment in low latitude or tropical regions, humans consume significant amounts of electrical energy in air conditioning, leading to substantial CO2 emissions. Passive daytime radiative cooling (PDRC) allows objects to cool down during the daytime without any energy consumption by dissipating heat through the atmospheric transparency window (8-13 µm) to outer space, which has garnered significant attention. However, the practical applications of common PDRC materials are hindered by their poor optical selectivity and high-reflective silver backing. Additionally, the availability of artificial photon emitters with complex structures and excellent performance is also limited by their high cost. Herein, a novel multilayer prismatic photonic metamaterial film without any silver reflector, easily scalable and produced by a roll-to-roll method is demonstrated, which exhibits ≈96.4% sunlight reflectance (0.3-2.5 µm) and ≈97.2% emissivity in mid-infrared (IR) (8-13 µm). At an average solar intensity of ≈920 W m-2 , it is on average 6.8 °C below ambient temperature during the day and theoretically yields a radiative cooling power of 88.9 W m-2 . Furthermore, the film exhibits excellent hydrophobicity, superior flexibility, and robust mechanical strength, providing an attractive and viable pathway for practical applications addressing the pressing challenges of climate and energy issues.

Redox Homeostasis Disruptors Based on Metal-Phenolic Network Nanoparticles for Chemo/Chemodynamic Synergistic Tumor Therapy through Activating Apoptosis and Cuproptosis.

The combination of chemo/chemodynamic therapy is a promising strategy for improving antitumor efficacy. Herein, metal-phenolic network nanoparticles (NPs) self-assembled from copper ions and gallic acid (Cu-GA) are developed to evoke apoptosis and cuproptosis for synergistic chemo/chemodynamic therapy. The Cu-GA NPs are biodegraded in response to the highly expressed glutathione (GSH) in tumor cells, resulting in the simultaneous release of Cu+ and GA. The intracellular GSH content is dramatically reduced by the released GA, rendering the tumor cells incapable of scavenging reactive oxygen species (ROS) and more susceptible to cuproptosis. Meanwhile, ROS levels within the tumor cells are significantly increased by the Fenton-like reaction of released Cu+ , which disrupts redox homeostasis and achieves apoptosis-related chemodynamic therapy. Moreover, massive accumulation of Cu+ in the tumor cells further induces aggregation of lipoylated dihydrolipoamide S-acetyltransferase and downregulation of iron-sulfur cluster protein, activating cuproptosis to enhance the antitumor efficacy of Cu-GA NPs. The experiments in vivo further demonstrate that Cu-GA NPs exhibited the excellent biosafety and superior antitumor capacity, which can efficiently inhibit the growth of tumors due to the activation by the tumor specific GSH and hydrogen peroxide. These Cu-based metal-phenolic network NPs provide a potential strategy to build up efficient and safe cancer therapy.

Transcriptomic Analysis of mRNA Expression Profiles in the Microglia of Mouse Brains Infected with Rabies Viruses of Varying Virulence.

Rabies is a lethal encephalitis caused by the rabies virus (RABV) with a fatality rate near 100% after the onset of clinical symptoms in humans and animals. Microglia are resident immune cells in the central nervous system. Few studies have been conducted on the functional role of microglia in RABV infection. Here, we performed a transcriptomic analysis of mRNA expression profiles in the microglia of mouse brains intracerebrally infected with RABV. We successfully isolated single microglial cells from the mouse brains. The survival rate of dissociated microglial cells was 81.91%-96.7%, and the purity was 88.3%. Transcriptomic analysis revealed 22,079 differentially expressed mRNAs identified in the microglia of mouse brains infected with RABV strains (rRC-HL, GX074, and CVS-24) of varying virulence at 4 and 7 days post-infection (dpi) compared to the control group. The numbers of DEGs versus the control at 4 and 7 dpi in mice infected with rRC-HL, GX074, and CVS-24 were 3622 and 4590, 265 and 4901, and 4079 and 6337. The GO enrichment analysis showed that response to stress, response to external stimulus, regulation of response to stimulus, and immune system process were abundant during RABV infection. The KEGG analysis indicated that the Tlr, Tnf, RIG-I, NOD, NF-κB, MAPK, and Jak-STAT signaling pathways were involved in RABV infection at both 4 and 7 dpi. However, some phagocytosis and cell signal transduction processes, such as endocytosis, p53, phospholipase D, and oxidative phosphorylation signaling pathways, were only expressed at 7 dpi. The involvement of the Tnf and Tlr signaling pathways prompted us to construct a protein-protein interaction (PPI) network of these pathways. The PPI revealed 8 DEGs, including Mmp9, Jun, Pik3r1, and Mapk12. Notably, Il-1b interacted with Tnf and Il-6 with combined scores of 0.973 and 0.981, respectively. RABV causes significant changes in mRNA expression profiles in the microglia in mice. 22,079 differentially expressed mRNAs were identified in the microglia of mice infected with RABV strains of varying virulence at 4 and 7 dpi. The DEGs were evaluated using GO, KEGG, and PPI network analysis. Many immune pathways were up-regulated in RABV-infected groups. The findings will help elucidate the microglial molecular mechanisms of cellular metabolism dysregulated by RABV and may provide important information for investigating RABV pathogenesis and therapeutic methods.

Preparation of Densified Fine-Grain High-Frequency MnZn Ferrite Using the Cold Sintering Process.

The densified MnZn ferrite ceramics were prepared using the cold sintering process under pressure, with an acetate ethanol solution used as the transient solvent. The effects of the transient solvent, the pressure and annealing temperature on the density, and the micromorphology and magnetic properties of the sintered MnZn ferrites were studied. The densified MnZn ferrite was obtained using the cold sintering process and its relative density reached up to 85.4%. The transient solvent and high pressure are essential to the cold sintering process for MnZn ferrite. The annealing treatment is indispensable in obtaining the sample with the higher density. The relative density was further increased to 97.2% for the sample annealed at 950 °C for 6 h. The increase in the annealing temperature reduces the power loss at high frequencies.

Integrative metabolomic and transcriptomic analysis reveals difference in glucose and lipid metabolism in the longissimus muscle of Luchuan and Duroc pigs.

Luchuan pig, an obese indigenous Chinese porcine breed, has a desirable meat quality and reproductive capacity. Duroc, a traditional western breed, shows a faster growth rate, high feed efficiency and high lean meat rate. Given the unique features these two porcine breeds have, it is of interest to investigate the underlying molecular mechanisms behind their distinctive nature. In this study, the metabolic and transcriptomic profiles of longissimus dorsi muscle from Duroc and Luchuan pigs were compared. A total of 609 metabolites were identified, 77 of which were significantly decreased in Luchuan compared to Duroc, and 71 of which were significantly elevated. Most differentially accumulated metabolites (DAMs) upregulated in Luchuan were glycerophospholipids, fatty acids, oxidized lipids, alcohols, and amines, while metabolites downregulated in Luchuan were mostly amino acids, organic acids and nucleic acids, bile acids and hormones. From our RNA-sequencing (RNA-seq) data we identified a total of 3638 differentially expressed genes (DEGs), 1802 upregulated and 1836 downregulated in Luchuan skeletal muscle compared to Duroc. Combined multivariate and pathway enrichment analyses of metabolome and transcriptome results revealed that many of the DEGs and DAMs are associated with critical energy metabolic pathways, especially those related to glucose and lipid metabolism. We examined the expression of important DEGs in two pathways, AMP-activated protein kinase (AMPK) signaling pathway and fructose and mannose metabolism, using Real-Time Quantitative Reverse Transcription PCR (qRT-PCR). Genes related to glucose uptake, glycolysis, glycogen synthesis, fatty acid synthesis (PFKFB1, PFKFB4, MPI, TPI1, GYS1, SLC2A4, FASN, IRS1, ULK1) are more activated in Luchuan, while genes related to fatty acid oxidation, cholesterol synthesis (CPT1A, HMGCR, FOXO3) are more suppressed. Energy utilization can be a decisive factor to the distinctive metabolic, physiological and nutritional characteristics in skeletal muscle of the two breeds we studied. Our research may facilitate future porcine breeding projects and can be used to reveal the potential molecular basis of differences in complex traits between various breeds.

Theoretical exploration of noncovalent interactions in Sc2C2@C2n (n = 40, 41, and 42)⊂[12]CPP, PF[12]CPP.

The encapsulation of fullerenes by carbon nanorings has gained increasing attention because of the unique molecular structure and special properties of the formed complexes. The host-guest interactions between the fullerenes and the carbon nanorings can influence the metal ion orientation and the molecular electronic structure. In this study, we hooped a series of carbide cluster metallofullerenes, namely Sc2C2@C2v(5)-C80, Sc2C2@C3v(8)-C82, and Sc2C2@D2d(23)-C84, with molecular carbon nanorings of [12]cycloparaphenylene ([12]CPP) and perfluoro[12]cycloparaphenylene (PF[12]CPP). The formed complexes were computationally studied via dispersion-corrected density functional theory calculations. The results showed that the deformation rate of PF[12]CPP after the formation of the fullerene-containing complexes was significantly smaller than that of [12]CPP. The binding energy and thermodynamic information showed that PF[12]CPP was more suitable for fullerene encapsulation. Moreover, charge population analysis showed that PF[12]CPP transferred more electrons to Sc2C2@C2n (n = 40, 41, and 42) compared with [12]CPP. Energy decomposition and real-space function analyses of host-guest interactions revealed the characteristics and nature of the noncovalent interactions in the supramolecules. These results provide theoretical support for the study of host-guest systems based on metallofullerenes.

Low Core Losses of Fe-Based Soft Magnetic Composites with an Zn-O-Si Insulating Layer Obtained by Coupling Synergistic Photodecomposition.

The major method used to reduce the magnetic loss of soft magnetic composites (SMCs) is to coat the magnetic powder with an insulating layer, but the permeability is usually sacrificed in the process. In order to achieve a better balance between low losses and high permeability, a novel photodecomposition method was used in this study to create a ZnO insulating layer. The effect of the concentration of diethyl zinc on the formation of a ZnO insulating film by photodecomposition was studied. The ZnO film was best formed with a diethyl zinc n-hexane solution at a concentration of around 0.40 mol/L. Combined with conventional coupling treatment processes, a thin and dense insulating layer was coated on the surface of iron powder in situ. Treating the iron powder before coating by photodecomposition led to a synergistic effect, significantly reduced core loss, and the effective permeability only decreased slightly. An iron-based soft magnetic composite with a loss value of 124 kW/m3 and an effective permeability of 107 was obtained at the frequency of 100 kHz and a magnetic field intensity of 20 mT.

Injection Molding and Sintering of ZrO2 Ceramic Powder Modified by a Zirconate Coupling Agent.

Ceramic injection molding is a near-net shape-processing technique, producing ceramic components with low tooling costs and complex shapes. In this paper, ZrO2 ceramics with high loading content in the green part were prepared by powder modification using zirconate coupling agent, injection molding and sintering, which benefited decreasing the usage of binders and deformation of ceramics. The rheological characteristics of feedstocks, densities, microstructures and mechanical properties of green and sintered parts with the different coupling media and sintering temperatures were studied. The results showed that the addition of a zirconate coupling agent with ethanol medium obviously increased the flowability of feedstocks and benefited achieving the green parts with high powder loading (86.5 wt.%) and bending strength (12.9 MPa) and the final unbroken ceramics. In addition, the sintering temperatures from 1500-1575 °C had no significant effects on the density, hardness, and surface morphology of the ceramic samples. However, the bending strength increased and some large grains with transgranular fracture occurred on the fractural surface at the sintering temperature of 1575 °C.

Profiling and Functional Analysis of long non-coding RNAs in yak healthy and atretic follicles.

Yak is the livestock on which people live in plateau areas, but its fecundity is low. Follicular development plays a decisive role in yak reproductive performance. As an important regulatory factor, the expression of long non-coding RNA (lncRNAs) in yak follicular development and its regulatory mechanism remains unclear. To explore the differentially expressed lncRNAs between healthy and atretic follicular in yaks. We used RNA-seq to construct lncRNA, miRNA, and mRNA expression profiles in yak atretic and healthy follicles, and the RNA sequence results were identified by qPCR. In addition, the correlation of lncRNA and targeted mRNA was also analyzed by Starbase software. Moreover, lncRNA/miRNA/mRNA networks were constructed by Cytoscape software, and the network was verified by dual-luciferase analysis. A total of 682 novel lncRNAs, 259 bta-miRNAs, and 1704 mRNAs were identified as differentially expressed between healthy and atretic follicles. Among them, 135 mRNAs were positively correlated with lncRNA expression and 97 were negatively correlated, which may be involved in the yak follicular development. In addition, pathway enrichment analysis of differentially expressed lncRNA host genes by Kyoto Genome Encyclopedia (KEGG) showed that host genes were mainly involved in hormone secretion, granulosa cell apoptosis, and follicular development. In conclusion, we identified a series of novel lncRNAs, constructed the lncRNA ceRNA regulatory network, and provided comprehensive resources for exploring the role of lncRNAs in yak ovarian follicular development.

2-Mercaptoethanol promotes porcine oocyte maturation in vitro by maintaining autophagy homeostasis.

2-Mercaptoethanol (2-ME) is often used as an antioxidant to optimize culture systems for in vitro oocyte maturation in livestock. However, the relationship between 2-ME and autophagy has not yet been elucidated. In this study, we hypothesized that 2-ME can promote porcine oocyte maturation in vitro by maintaining autophagy homeostasis. To test this hypothesis, we explored the effects of 2-ME on the maturation of porcine oocytes exposed to an autophagy activator (rapamycin) or an autophagy inhibitor (3-methyladenine, i.e., 3-MA) in vitro. Rapamycin-induced autophagy over-activation significantly increased autophagy- and apoptosis-related gene expression, oxidative stress, apoptosis rates, abnormal mitochondrial redistribution, and significantly decreased oocyte first polar body extrusion (PBE) rates, spindle/chromosome integrity and developmental competence. 3-MA-mediated autophagy inhibition exerted similar effects on all these parameters except the expression of genes that promote autophagy and inhibit apoptosis. Importantly, 2-ME supplementation significantly attenuated the detrimental effects of rapamycin and 3-MA. Interestingly, we observed that 44 h of coincubation with rapamycin/3-MA and 2-ME restored autophagy homeostasis in vitro. In conclusion, our study confirmed that 2-ME promotes porcine oocyte maturation and embryo development in vitro by maintaining autophagy homeostasis and lays a foundation for further research on the underlying mechanism.

Allicin protects porcine oocytes against LPS-induced defects during maturation in vitro.

Lipopolysaccharide (LPS), the main virulence factor of gram-negative bacteria, severely impairs the function of the female reproductive system. It has especially harmful effects on oocytes and subsequent embryonic development. The use of active plant substances to ameliorate the damage caused by LPS exposure is a strategy worthy of attention. In this study, porcine oocytes were used to investigate the protective effects and underlying mechanisms of allicin, an extract derived from garlic, on LPS-exposed oocytes in vitro. Our data indicated that supplementation with 1 µM allicin significantly attenuated the LPS-mediated reductions in the first polar body extrusion rate and the subsequent blastocyst formation rate. Allicin also mitigated the abnormalities in spindle assembly, chromosome alignment, actin polymerization, and cortical granule distribution caused by LPS exposure. Furthermore, allicin restored reactive oxygen species (ROS), early apoptosis and autophagy to normal physiological levels in LPS-exposed oocytes. In conclusion, our findings confirm that allicin can protect oocytes against LPS-induced damage. The results of this study will help promote the application of plant-derived bioactive substances to ameliorate oocyte maturation defects.

Carbothermal Reduction Synthesis of Aluminum Nitride from Al(OH)3/C/PVB Slurries Prepared by Three-Roll Mixing.

Polyvinyl butyral (PVB) was used in the Al(OH)3/carbon black/ethanol slurries by the three-roll mixing to prepare AlN powder using the carbothermal reduction-nitridation (CRN) process in the experiments. The effects of PVB addition on the synthesis of AlN powder were studied by viscosity, tap density, XRD, SEM and TG measurements. The results showed that the PVB layer covering on the surface of Al(OH)3 particles reduced the viscosity of Al(OH)3/carbon/ethanol slurry and increased the dispersion homogeneity of Al(OH)3/carbon raw powder. The tap densities of the Al(OH)3/carbon mixtures after three-roll milling could be increased with the increase in PVB addition. In the CRN process, most of the PVB covering Al(OH)3 particles evaporated and supplied the passage for nitrogen removal to the particles. Based on the experimental data, the role of PVB on the mixing and CRN process was discussed.

H2O2-independent chemodynamic therapy initiated from magnetic iron carbide nanoparticle-assisted artemisinin synergy.

Chemodynamic therapy (CDT) is a booming technology that utilizes Fenton reagents to kill tumor cells by transforming intracellular H2O2 into reactive oxygen species (ROS), but insufficient endogenous H2O2 makes it difficult to attain satisfactory antitumor results. In this article, a H2O2-free CDT technique with tumor-specificity is developed by using pH-sensitive magnetic iron carbide nanoparticles (PEG/Fe2C@Fe3O4 NPs) to trigger artemisinin (ART) to in situ form ROS. ART-loaded PEG/Fe2C@Fe3O4 NPs are fabricated for the enormous release of Fe2+ ions induced by the acidic conditions of the tumor microenvironment after magnetic-assisted tumor enrichment, which results in the rapid degradation of the PEG/Fe2C@Fe3O4 NPs and release of ART once endocytosed into tumor cells. In situ catalysis reaction between the co-released Fe2+ ions and ART generates toxic ROS and then induces apoptosis of tumor cells. Both in vitro and in vivo experiments demonstrate that the efficient Fe-enhanced and tumor-specific CDT efficacy for effective tumor inhibition based on ROS generation. This work provides a new direction to improve CDT efficacy based on H2O2-independent ROS generation.

Magnetic Reactive Oxygen Species Nanoreactor for Switchable Magnetic Resonance Imaging Guided Cancer Therapy Based on pH-Sensitive Fe5C2@Fe3O4 Nanoparticles.

Reactive oxygen species (ROS) are crucial molecules in cancer therapy. Unfortunately, the therapeutic efficiency of ROS is unsatisfactory in clinic, primarily due to their rigorous production conditions. By taking advantage of the intrinsic acidity and overproduction of H2O2 in the tumor environment, we have reported an ROS nanoreactor based on core-shell-structured iron carbide (Fe5C2@Fe3O4) nanoparticles (NPs) through the catalysis of the Fenton reaction. These NPs are able to release ferrous ions in acidic environments to disproportionate H2O2 into •OH radicals, which effectively inhibits the proliferation of tumor cells both in vitro and in vivo. The high magnetization of Fe5C2@Fe3O4 NPs is favorable for both magnetic targeting and T2-weighted magnetic resonance imaging (MRI). Ionization of these NPs simultaneously decreases the T2 signal and enhances the T1 signal in MRI, and this T2/T1 switching process provides the visualization of ferrous ions release and ROS generation for the supervision of tumor curing. These Fe5C2@Fe3O4 NPs show great potential in endogenous environment-excited cancer therapy with high efficiency and tumor specificity and can be guided further by MRI.

Self-Assemblies of Acicular Hollow Fe/C Nanostructures.

Self-assemblies of acicular hollow Fe/C structures were synthesized using D-glucose monohydrate and ferric chloride as precursors by a simple hydrothermal process followed by carbonization at 800 °C. The self-assembled structures with an overall diameter of 15~20 µm composed of radially formed hollow needles from a central core with an average diameter of ca. 1 µm and a length up to 10 µm. The end of the needles was revealed to be a awl shape with a hollow structure formed during the self-assembly process and the subsequent heat treatment. The hollow structure was probably caused by the Kirkendall effect at 800 °C. The materials exhibit ferromagnetic characteristic with saturation magnetization (Ms), remanent magnetization (Mr), and coercivity (Hc) of 22.2 emu/g, 3 emu/g, and 151.22 Oe, respectively, with Ms much lower than that of Fe3O4.

Preparation and microwave absorbing properties of carbon/cobalt ferromagnetic composites.

Carbon/cobalt ferromagnetic light composites with high performance of microwave absorbing properties were prepared by hydrothermal method using starch and hollow cobalt ferrites. It was concluded that after carbonization the spinel structure ferrites changed to Co3Fe7 alloys and the temperature of graphitization was significantly decreased for the catalytic of CoFe2O4/Co3Fe7. The increase of carbon content, and exist of CoFe2O4/Co3Fe7 heightened the microwave absorbing properties. Electromagnetic parameters were tested with 40% of the titled materials and 60% of paraffin wax composites by using HP8722ES vector network analyzer. The reflection was also simulated through transmission line theory. The microwave absorbers exhibited a maximum reflection loss -43 dB and the electromagnetic wave absorption less than -10 dB was found to exceed 3.0 GHz between 11.6 GHz and 15 GHz for an absorber thickness of 2 mm.