Increasing evidence suggests that long noncoding RNAs (lncRNAs) play crucial roles in various biological processes. However, little is known about the effects of lncRNAs on autophagy. Here we report ...that a lncRNA, termed cardiac autophagy inhibitory factor (CAIF), suppresses cardiac autophagy and attenuates myocardial infarction by targeting p53-mediated myocardin transcription. Myocardin expression is upregulated upon H
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and ischemia/reperfusion, and knockdown of myocardin inhibits autophagy and attenuates myocardial infarction. p53 regulates cardiomyocytes autophagy and myocardial ischemia/reperfusion injury by regulating myocardin expression. CAIF directly binds to p53 protein and blocks p53-mediated myocardin transcription, which results in the decrease of myocardin expression. Collectively, our data reveal a novel CAIF-p53-myocardin axis as a critical regulator in cardiomyocyte autophagy, which will be potential therapeutic targets in treatment of defective autophagy-associated cardiovascular diseases.
Characterizing the long-term nanometer-scale interactions between lysosomes and mitochondria in live cells is essential for understanding their functions but remains challenging due to limitations of ...the existing fluorescent probes. Here, we develop cell-permeable organic fluorescent probes for lysosomes with excellent specificity and high photostability. We also use an existing Atto 647N dye with high brightness and excellent photostability to achieve specific labeling of mitochondria in live cells. Using these probes, we obtain dual-color structured illumination microscopy (SIM) images of dynamic physical lysosome-mitochondrion interactions in live cells at an ~90-nm resolution over a long time course of ~13 min. We successfully record the consecutive dynamic processes of lysosomal fusion and fission, as well as four types of physical lysosome-mitochondrion interactions by super-resolution imaging. Our probes provide an avenue for understanding the functions and the dynamic interplay of lysosomes and mitochondria in live cells.
Dysregulated autophagy is associated with many pathological disorders such as cardiovascular diseases. Emerging evidence has suggested that circular RNAs (circRNAs) have important roles in some ...biological processes. However, it remains unclear whether circRNAs participate in the regulation of autophagy. Here we report that a circRNA, termed autophagy-related circular RNA (ACR), represses autophagy and myocardial infarction by targeting Pink1-mediated phosphorylation of FAM65B. ACR attenuates autophagy and cell death in cardiomyocytes. Moreover, ACR protects the heart from ischemia/reperfusion (I/R) injury and reduces myocardial infarct sizes. We identify Pink1 as an ACR target to mediate the function of ACR in cardiomyocyte autophagy. ACR activates Pink1 expression through directly binding to Dnmt3B and blocking Dnmt3B-mediated DNA methylation of Pink1 promoter. Pink1 suppresses autophagy and Pink1 transgenic mice show reduced myocardial infarction sizes. Further, we find that FAM65B is a downstream target of Pink1 and Pink1 phosphorylates FAM65B at serine 46. Phosphorylated FAM65B inhibits autophagy and cell death in the heart. Our findings reveal a novel role for the circRNA in regulating autophagy and ACR-Pink1-FAM65B axis as a regulator of autophagy in the heart will be potential therapeutic targets in treatment of cardiovascular diseases.
Shuxuening injection (SXNI), a popular herbal medicine, is an extract of Ginkgo biloba leaves (GBE), and is used to treat ischemic stroke (IS) in China. However, its specific active ingredients and ...molecular mechanisms in IS remain unclear.
Aim of the study: The purpose of the research is to identify the main active ingredients in GBE and explore its molecular mechanisms in the treatment of IS.
The main active components of GBE were discerned through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis (TCMSP), Traditional Chinese Medicine Integrated Database (TCMID), Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM) database, and absorption, distribution, metabolism and excretion (ADME) analysis. The targets related to IS were obtained using Genecards, Online Mendelian Inheritance in Man (OMIM), Therapeutic Target Database (TTD), and Disgenet. We discovered an intersection of genes. Subsequently, protein-protein interaction (PPI) networks were constructed with Cytoscape 3.7.1 and the String database. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to analyze the intersection of targets via the Database for Annotation, Visualization, and Integrated Discovery (DAVID) 6.8. Built on the above analysis, we made a Compound-Target-Pathway (C-T-P) network. Autodock Vina was used for molecular docking analysis. Maestro 11.9 was used to calculate the root-mean-square deviation (RMSD). Animal experiments were performed to verify the core targets. Triphenyl tetrazolium chloride (TTC) staining was used to calculate the infarct volume in rats. Hematoxylin-eosin (HE) staining was employed to observe the morphology of hippocampal neuron cells. RT-qPCR was applied to detect relative mRNA levels, and protein expression was determined using Western blotting.
Molecular docking showed that PTGS2, NOS3 and CASP3 docked with small molecule compounds. According to RT-qPCR and Western blotting, mRNA and protein expression of PTGS2 and CASP3 were up-regulated (P < 0.05), and mRNA and protein levels of NOS3 were down-regulated (P < 0.05).
SXNI can treat IS through multiple targets and routes, and reduce the apoptosis of neuron cells in brain tissue by inhibiting inflammation and regulating the level of oxidative stress, thereby protecting rats brain tissue.
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Particle swarm optimization (PSO) relies on its learning strategy to guide its search direction. Traditionally, each particle utilizes its historical best experience and its neighborhood's best ...experience through linear summation. Such a learning strategy is easy to use, but is inefficient when searching in complex problem spaces. Hence, designing learning strategies that can utilize previous search information (experience) more efficiently has become one of the most salient and active PSO research topics. In this paper, we proposes an orthogonal learning (OL) strategy for PSO to discover more useful information that lies in the above two experiences via orthogonal experimental design. We name this PSO as orthogonal learning particle swarm optimization (OLPSO). The OL strategy can guide particles to fly in better directions by constructing a much promising and efficient exemplar. The OL strategy can be applied to PSO with any topological structure. In this paper, it is applied to both global and local versions of PSO, yielding the OLPSO-G and OLPSO-L algorithms, respectively. This new learning strategy and the new algorithms are tested on a set of 16 benchmark functions, and are compared with other PSO algorithms and some state of the art evolutionary algorithms. The experimental results illustrate the effectiveness and efficiency of the proposed learning strategy and algorithms. The comparisons show that OLPSO significantly improves the performance of PSO, offering faster global convergence, higher solution quality, and stronger robustness.
Efficient charge separation and utilization are critical factors in photocatalysis. Herein, it is demonstrated that the complete spatial separation of oxidation and reduction cocatalysts enhances the ...efficacy of charge separation and surface reaction. Specifically, a Pt@NH2‐UiO‐66@MnOx (PUM) heterostructured photocatalyst with Pt and MnOx as cocatalysts is designed for the optimization of the NH2‐UiO‐66 photocatalyst. Compared with the pristine NH2‐UiO‐66, Pt@NH2‐UiO‐66 (PU), and NH2‐UiO‐66@MnOx (UM) samples, the PUM sample exhibits the highest hydrogen production activity. As cocatalysts, Pt favors trapping of electrons, while MnOx tends to collect holes. Upon generation from NH2‐UiO‐66, electrons and holes flow inward and outward of the metal–organic framework photocatalyst, accumulating on the corresponding cocatalysts, and then take part in the redox reactions. The PUM photocatalyst greatly prolongs the lifetime of the photogenerated electrons and holes, which favors the electron–hole separation. Furthermore, the PUM sample facilitates overall water splitting in the absence of sacrificial agents, thereby demonstrating its potential as a modification method of MOF‐type semiconductors for the overall water‐splitting reaction.
A heterostructured photocatalyst, Pt@NH2‐UiO‐66@MnOx (PUM), is constructed based on the rational optimization of NH2‐UiO‐66 with spatially separated Pt and MnOx nanoparticles as cocatalysts. The optimization results in a long lifetime of the photogenerated electrons and holes in the composite, which can drive overall water splitting in the absence of sacrificial agents.
Long-term visualization of the dynamic interactions between intracellular structures throughout the three-dimensional space of whole live cells is essential to better understand their functions, but ...this task remains challenging due to the limitations of existing three-dimensional fluorescence microscopy techniques, such as an insufficient axial resolution, low volumetric imaging rate and photobleaching. Here, we present the combination of a progressive deep-learning super-resolution strategy with a double-ring-modulated selective plane illumination microscopy design capable of visualizing the dynamics of intracellular structures in live cells for hours at an isotropic spatial resolution of roughly 100 nm in three dimensions at speeds up to roughly 17 Hz. Using this approach, we reveal the complex spatial relationships and interactions between endoplasmic reticulum (ER) and mitochondria throughout live cells, providing new insights into ER-mediated mitochondrial division. We also examined the motion of Drp1 oligomers involved in mitochondrial fission and revealed the dynamic interactions between Drp1 and mitochondria in three dimensions.
The adult mammalian cardiomyocytes lose their proliferative capacity, which is responsible for cardiac dysfunction and heart failure following injury. The molecular mechanisms underlying the ...attenuation of adult cardiomyocyte proliferation remain largely unknown. Because long noncoding RNAs (lncRNAs) have a critical role in the development of cardiovascular problems, we investigated whether lncRNAs have any role in the regulation of cardiomyocyte proliferation and cardiac repair.
Using bioinformatics and initial analysis, we identified an lncRNA, named CPR (cardiomyocyte proliferation regulator), that has a potential regulatory role in cardiomyocyte proliferation. For in vivo experiments, we generated CPR knockout and cardiac-specific CPR-overexpressing mice. In isolated cardiomyocytes, we used adenovirus for silencing (CPR-small interfering RNA) or overexpressing CPR. To investigate the mechanisms of CPR function in cardiomyocyte proliferation, we performed various analyses including quantitative reverse transcription-polymerase chain reaction, Western blot, histology, cardiac function (by echocardiography), transcriptome analyses (microarray assay), RNA pull-down assay, and chromatin immunoprecipitation assay.
CPR level is comparatively higher in the adult heart than in the fetal stage. The silencing of CPR significantly increased cardiomyocyte proliferation in postnatal and adult hearts. Moreover, CPR deletion restored the heart function after myocardial injury, which was evident from increased cardiomyocyte proliferation, improvement of myocardial function, and reduced scar formation. In contrast, the neonatal cardiomyocyte proliferation and cardiac regeneration were remarkably suppressed in CPR-overexpressing mice or adeno-associated virus serotype 9-CPR-overexpressing heart. These results indicate that CPR acts as a negative regulator of cardiomyocyte proliferation and regeneration. Next, we found that CPR targets minichromosome maintenance 3, an initiator of DNA replication and cell cycle progression, to suppress cardiomyocyte proliferation. CPR silenced minichromosome maintenance 3 expression through directly interacting and recruiting DNMT3A to its promoter cysteine-phosphate-guanine sites, as evident from decreased minichromosome maintenance 3 promoter methylation and increased minichromosome maintenance 3 expression in CPR knocked-down cardiomyocytes and CPR knockout mouse heart. These results were confirmed in CPR-overexpressing cardiomyocytes and CPR-overexpressing mouse heart.
Together, our findings identified that CPR is a suppressor of cardiomyocyte proliferation and indicated that lncRNAs take part in the regulation of cardiomyocyte proliferation and cardiac repair. Our study provides an lncRNA-based therapeutic strategy for effective cardiac repair and regeneration.
A novel enzyme-responsive supramolecular polysaccharide assembly composed of disulfide linked adamantane-naphthalimide fluorescent camptothecin prodrug (AdaCPT) and β-CD modified hyaluronic acid ...(HACD) was constructed, possessing low cellular cytotoxicity and exhibiting targeted cellular imaging and controlled drug release at specific sites while providing a concurrent means for the real-time tracking of drug delivery.
Summary
The plant family 1 UDP‐glycosyltransferases (UGTs) are the biggest GT family in plants, which are responsible for transferring sugar moieties onto a variety of small molecules, and control ...many metabolic processes; however, their physiological significance in planta is largely unknown. Here, we revealed that two Arabidopsis glycosyltransferase genes, UGT79B2 and UGT79B3, could be strongly induced by various abiotic stresses, including cold, salt and drought stresses. Overexpression of UGT79B2/B3 significantly enhanced plant tolerance to low temperatures as well as drought and salt stresses, whereas the ugt79b2/b3 double mutants generated by RNAi (RNA interference) and CRISPR‐Cas9 strategies were more susceptible to adverse conditions. Interestingly, the expression of UGT79B2 and UGT79B3 is directly controlled by CBF1 (CRT/DRE‐binding factor 1, also named DREB1B) in response to low temperatures. Furthermore, we identified the enzyme activities of UGT79B2/B3 in adding UDP‐rhamnose to cyanidin and cyanidin 3‐O‐glucoside. Ectopic expression of UGT79B2/B3 significantly increased the anthocyanin accumulation, and enhanced the antioxidant activity in coping with abiotic stresses, whereas the ugt79b2/b3 double mutants showed reduced anthocyanin levels. When overexpressing UGT79B2/B3 in tt18 (transparent testa 18), a mutant that cannot synthesize anthocyanins, both genes fail to improve plant adaptation to stress. Taken together, we demonstrate that UGT79B2 and UGT79B3, identified as anthocyanin rhamnosyltransferases, are regulated by CBF1 and confer abiotic stress tolerance via modulating anthocyanin accumulation.
Significance Statement
The physiological roles of most UDP‐glycosyltransferases (UGTs) are unknown. Here, we used overexpression and RNA interference or CRISPR‐Cas9 strategies to show that two anthocyanin rhamnosyltransferases play crucial roles in enhancing tolerance to cold, salt and drought.
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