miR-29b-1-5p exacerbates myocardial injury induced by sepsis in a mouse model by targeting TERF2.

Myocardial damage is a critical complication and a significant contributor to mortality in sepsis. MicroRNAs (miRNAs) have emerged as key players in sepsis pathogenesis. In this study, we explore the effect and mechanisms of miR-29b-1-5p on sepsis-induced myocardial damage. Sepsis-associated Gene Expression Omnibus datasets (GSE72380 and GSE29914) are examined for differential miRNAs. The mouse sepsis-induced cardiac injury was established by Lipopolysaccharide (LPS) or cecal ligation and puncture (CLP). LPS-treated HL-1 mouse cardiomyocytes simulate myocardial injury in vitro. miR-29b-1-5p is co-upregulated in both datasets and in cardiac tissue from sepsis mouse and HL-1 cell models. miR-29b-1-5p expression downregulation was achieved by antagomir transduction and confirmed by real-time quantitative reverse transcription PCR. Survival analysis and echocardiography examination show that miR-29b-1-5p inhibition improves mice survival cardiac function in LPS- and CLP-induced sepsis mice. Hematoxylin and eosin and Masson's trichrome staining and Immunohistochemistry analysis of mouse myocardial α-smooth muscle actin show that miR-29b-1-5p inhibition reduces myocardial tissue injury and fibrosis. The inflammatory cytokines and cardiac troponin I (cTnI) levels in mouse serum and HL-1 cells are also decreased by miR-29b-1-5p inhibition, as revealed by enzyme-linked immunosorbent assay. The expressions of autophagy-lysosomal pathway-related and apoptosis-related proteins in the mouse cardiac tissues and HL-1 cells are evaluated by western blot analysis. The sepsis-induced activation of the autophagy-lysosomal pathway and apoptosis are also reversed by miR-29b-1-5p antagomir. MTT and flow cytometry measurement further confirm the protective role of miR-29b-1-5p antagomir in HL-1 cells by increasing cell viability and suppressing cell apoptosis. Metascape functionally enriches TargetScan-predicted miR-29b-1-5p target genes. TargetScan prediction and dual luciferase assay validate the targeting relationship between miR-29b-1-5p and telomeric repeat-binding factor 2 (TERF2). The expression and function of TERF2 in HL-1 cells and mice are also evaluated. MiR-29b-1-5p negatively regulates the target gene TERF2. TERF2 knockdown partly restores miR-29b-1-5p antagomir function in LPS-stimulated HL-1 cells. In summary, miR-29b-1-5p targetedly inhibits TERF2, thereby enhancing sepsis-induced myocardial injury.

Enhancement of Hydration Activity and Microstructure Analysis of γ-C2S.

This paper investigated the combined effect of chemical activators and nano-SiO2 on the hydration reaction and the microstructure of γ-C2S. The hydration reaction of γ-C2S slurry activated with chemical activators (NaHCO3, NaOH, K2CO3, and KOH at 1 mol/L) was enhanced by 1% nano-SiO2. The hydrate reaction rate was determined by isothermal calorimetry, and the hydrated samples were characterized by XRD, TGA/DTG, SEM-EDS, and 29Si MAS/NMR. The results revealed a substantial enhancement in the hydration activity of γ-C2S due to the presence of the alkaline activator. Furthermore, nano-SiO2 did not alter the composition of γ-C2S hydration products, instead providing nucleation sites for the growth of hydration products. Incorporating nano-SiO2 promoted the formation of C-(R)-S-H gel with a low calcium-to-silica ratio and increased its polymerization levels, resulting in more favorable structures. Among all the activators used in this study, potassium salts had a better activation effect than sodium salts. After 28 days of curing, the degree of hydration reaction in the KC+Si group was 48% and about 37% for the NHC+Si group. Whereas, the KH+Si and NH+Si groups only reached approximately 20% after the same hydration duration.

Model for predicting prognosis and immunotherapy based on CD+8 T cells infiltration in neuroblastoma.

BACKGROUND: Neuroblastoma (NBL) is an extracranial malignant tumor in children deriving from the neural crest in the sympathetic nervous system. Although various immunotherapy interventions have made significant breakthroughs in many adult cancers, the efficacy of these immunotherapies was still limited in NBL. NBL has low immunogenicity which results in a lack of tumor-infiltrating T lymphocytes in the tumor microenvironment (TME). Moreover, tumor cells can wield many immune evasion strategies both in the TME and systemically to impede lymphocyte infiltration and activation. All these factors hamper the anti-tumor effects of CD8+ T cells during immunotherapy and the levels of infiltrating CD8+ T cells correlate with therapy response. MATERIALS AND METHODS: In this study, we utilized multidimensional bioinformatic methods to establish a risk model based on CD8+ T cells -related genes (CD8+ TRGs). RESULTS: We obtained 33 CD8+ TRGs with well-predictive ability for prognosis in both GSE49711 and E-MTAB-8248 cohorts. Then, 12 CD8+ TRGs including HK2, RP2, HPSE, ELL2, GFI1, SLC22A16, FCGR3A, CTSS, SH2D1A, RBP5, ATF5, and ADAM9 were finally identified for risk model construction and validation. This model revealed a stable performance in prognostic prediction of the overall survival (OS) and event-free survival (EFS) in patients with NBL. Additionally, our research indicated that the immune and stromal scores, immune-related pathways, immune cell infiltration, the expression of major histocompatibility complex (MHC) and immune checkpoint molecules, immunotherapy response, and drug susceptibility revealed significant differences between high and low-risk groups. CONCLUSIONS: According to our analyses, the constructed CD8+ TRGs-based risk model may be promising for the clinical prediction of anti-tumor therapy responses and prognoses in NBL.

Silencing MYOT Expression May Inhibit Autophagy in Human Skeletal Muscle Cells.

Muscle diseases are closely related to autophagy disorders. Studies of autophagy inhibition indicated the importance of autophagy in muscle regeneration, while activation of autophagy can restore muscle function in some myopathies. Previous studies have revealed that mutations in the MYOT gene may lead to several kinds of hereditary myopathies. However, whether the autophagy played a crucial role in hereditary myopathy caused by MYOT mutations was still not clear. In this study, we established the MYOT knockdown human skeletal muscle cell models (HSkMCs) by small interfering RNA. Real-time PCR and Western blot studies found that the expression of p62 and LC3B-II was decreased dramatically, which suggested that silencing MYOT expression may regulate the autophagy in HSkMCs. Further immunofluorescence study on Ad-mCherry-GFP-LC3B adenovirus transfection and monodansylcadaverine (MDC) staining revealed that knocking down the expression of MYOT may inhibit the autophagy. Next, we used the autophagy inducer Earle's balanced salt solution (EBSS) and late-autophagy inhibitor bafilomycin A1 (BAF A1) to treat the HSkMCs, respectively, and found that silencing MYOT expression can inhibit the activation of autophagy by EBSS and aggravate the inhibition of autophagy by BAF A1. Finally, we also found that silencing MYOT expression can downregulate the expression of ATG7 and ATG5, two important autophagy regulatory molecules. Hence, our study may first reveal that knocking down the expression of MYOT may inhibit the autophagy. Hereditary myopathies caused by MYOT mutations may partly result from the inhibition of autophagy in HSkMCs.

The Synergistic Effect of Ester-Ether Copolymerization Thixo-Tropic Superplasticizer and Nano-Clay on the Buildability of 3D Printable Cementitious Materials.

The shape retention ability of materials deposited layer by layer is called buildability, which is an indispensable performance parameter for successful 3D printable cementitious materials (3DPC). This study investigated the synergistic effect of nano-clay (NC) and thixotropic superplasticizer (TP) on the buildability of 3DPC. The rheological parameters and static yield stress are characterized by the rheology testing, the green strength is measured by a self-made pressure tester, and the fluidity is tested by flow table. Results indicate that NC significantly increases the growth rate of static yield stress and green strength and TP can improve the initial rheological parameters and fluidity, which ensures the initial stiffness and workability of printed materials. The mixture with 7‰ (by mass of cementitious materials) NC and 3‰ TP obtains excellent extrudability and buildability, due to the synergistic effect of NC and TP. Based on the rheology testing and specific printing experiments, a printable window with 1.0 Pa/s~2.0 Pa/s of the rate of static yield stress evolution over time (RST) or 170 mm~200 mm of fluidity is established. This work provides theorical support for the control and evaluation of rheological properties in 3DPC.

Effect of Structural Build-Up on Interlayer Bond Strength of 3D Printed Cement Mortars.

Understanding the relationship between the intrinsic characteristics of materials (such as rheological properties and structural build-up) and printability and controlling intrinsic characteristics of materials through additives to achieve excellent printability is vital in digital concrete additive manufacturing. This paper aims at studying the effects of material's structural build-up on the interlayer bond strength of 3DPC with different time gaps. Structural build-up can indirectly affect the interlayer bond strength by affecting the surface moisture of concrete. Based on the structural build-up of 3DPC, a new parameter, maximum operational time (MOT), is proposed, which can be considered as the limit of time gap to ensure high interlayer bond strength. Slump-retaining polycarboxylate superplasticizer (TS) slightly slows down the physical flocculation rate, but increases the maximum operational time of the cement paste. Nano clay significantly increases the sort-term structural build-up rate and has the function of internal curing and water retaining. Composite with nano-clay and TS can reduce the loss of surface moisture of 3D printed layers, prevent the formation of interface weak layer, and increase the interlayer bond strength between printed layers. This contribution can provide new insight into the design of 3D-printed ink with good extrudability, outstanding buildability, and excellent interlayer bond strength.

NBD-based fluorescent probes for separate detection of cysteine and biothiols via different reactivities.

NBD-based fluorescent probes that can separately detect cysteine and biothiols via different reactivities have been rationally designed and synthesized. The probes can be applied to kinetically distinguish cysteine from other biothiols at 30 min, and to detect all biothiols at 3 h in living cells.

Dual-quenching NBD-based fluorescent probes for separate detection of H2S and Cys/Hcy in living cells.

Biothiols are implicated in different physiological and pathological processes, while abnormal levels of biothiols are related to numerous diseases. It is essential to discriminatively detect these molecules and explore their inherent transformation in living biological samples. Herein, dual-reactive and dual-quenching fluorescent probes based on thiolysis of NBD (7-nitro-1,2,3-benzoxadiazole) thioether/ether/amine for separate detection of H2S and Cys/Hcy were rationally designed. Probe 2 was able to sense H2S (λex/em = 405/455 nm) and Cys/Hcy (λex/em = 470/550 nm) in the presence of GSH in aqueous buffer. Moreover, the probe not only could be applied for visualization of exogenous H2S and Cys in living cells but also successfully showcased the potential to verify the biosynthesis of endogenous H2S from Cys.

Continuum percolation of congruent overlapping polyhedral particles: Finite-size-scaling analysis and renormalization-group method.

The continuum percolation of randomly orientated overlapping polyhedral particles, including tetrahedron, cube, octahedron, dodecahedron, and icosahedron, was analyzed by Monte Carlo simulations. Two numerical strategies, (1) a Monte Carlo finite-size-scaling analysis and (2) a real-space Monte Carlo renormalization-group method, were, respectively, presented in order to determine the percolation threshold (e.g., the critical volume fraction ϕ_{c} or the critical reduced number density η_{c}), percolation transition width Δ, and correlation-length exponent ν of the polyhedral particles. The results showed that ϕ_{c} (or η_{c}) and Δ increase in the following order: tetrahedron < cube < octahedron < dodecahedron < icosahedron. In other words, both the percolation threshold and percolation transition width increase with the number of faces of the polyhedral particles as the shape becomes more "spherical." We obtained the statistical values of ν for the five polyhedral shapes and analyzed possible errors resulting in the present numerical values ν deviated from the universal value of ν=0.88 reported in literature. To validate the simulations, the corresponding excluded-volume bounds on the percolation threshold were obtained and compared with the numerical results. This paper has practical applications in predicting effective transport and mechanical properties of porous media and composites.

Chemical bonding and Cu diffusion at the Cu/Ta2N interface: a DFT study.

Ta2N is an effective diffusion barrier material to prevent undesired Cu diffusion in ultra-large scale integration circuits. Previous theoretical work has reported the interesting result that at the Cu/Ta2N interface the Cu layer preferentially bonded with the Ta layer but not the N layer of Ta2N. However, this result was calculated from largely lattice mismatched interface models. To confirm this theoretical result and unravel the cause of strong Cu-Ta bonding at the Cu/Ta2N interface, in this study density functional theory calculations, on the basis of super-cell models, were performed to investigate the Cu(111)/Ta2N(001) interface. We firstly calculated interface cohesive energies and confirmed that the Cu layer preferentially bonded with the Ta layer of Ta2N. Then, electronic structure calculations revealed that the chemical bonding of the Cu-Ta bond at the Cu(111)/Ta2N(001) interface was primarily covalent in character, providing a proper explanation for the close integration of the Cu layer and Ta layer. Lastly, Cu diffusion investigations revealed that Ta2N was able to effectively prevent Cu diffusion. Furthermore, we found that the N layer of Ta2N played the critical role in preventing Cu diffusion.

The interaction of NOS1AP, DISC1, DAOA, and GSK3B confers susceptibility of early-onset schizophrenia in Chinese Han population.

Although many major breakthrough had identificated potential susceptibility genes for schizophrenia, the aetiology of schizophrenia is still unknown. In the present study, we focused on the N-methyl-Daspartate receptors related genes nitric oxide synthase 1 adaptor gene (NOS1AP), disrupted in schizophrenia 1 gene (DISC1), d-amino acid oxidase activator gene (DAOA), and glycogen synthase kinase 3-beta gene (GSK3B). A family-based genetic association study (459 Han Chinese subjects in 153 nuclear families) using 3 single nucleotide polymorphisms in NOS1AP, 2 in DISC1, 1 in DAOA and 1 in GSK3B was conducted. We found rs12742393 have just positive trend with schizophrenia (SCZ) (p=0.07) after FDR correction. NOS1AP mRNA and serum levels were significantly elevated in SCZ patients (p<0.001; p<0.001) compared with healthy control. However, expression Quantitative Trait Loci (eQTL) analysis have demonstrated that rs12742393 genotype were not significantly associated with the NOS1AP mRNA expression. GMDR identified a significant seven-locus interaction model involving (NOS1AP-rs348624, rs12742393, rs1415263, DISC1-rs821633, rs1000731, DAOA-rs2391191and GSK3B- rs6438552) with a good testing accuracy (0.72). Our finding suggested statistically significant role of interaction of NOS1AP, DISC1, DAOA, and GSK3B polymorphisms (NOS1AP-rs348624, rs12742393, rs1415263, DISC1-rs821633, rs1000731, DAOA-rs2391191and GSK3B-rs6438552) in EOS susceptibility.

Comparative proteomic analysis of eggplant (Solanum melongena L.) heterostylous pistil development.

Heterostyly is a common floral polymorphism, but the proteomic basis of this trait is still largely unexplored. In this study, self- and cross-pollination of L-morph and S-morph flowers and comparison of embryo sac development in eggplant (Solanum melongena L.) suggested that lower fruit set from S-morph flowers results from stigma-pollen incompatibility. To explore the molecular mechanism underlying heterostyly development, we conducted isobaric tags for relative and absolute quantification (iTRAQ) proteomic analysis of eggplant pistils for L- and S-morph flowers. A total of 5,259 distinct proteins were identified during heterostyly development. Compared S-morph flowers with L-morph, we discovered 57 and 184 differentially expressed proteins (DEPs) during flower development and maturity, respectively. Quantitative real time polymerase chain reactions were used for nine genes to verify DEPs from the iTRAQ approach. During flower development, DEPs were mainly involved in morphogenesis, biosynthetic processes, and metabolic pathways. At flower maturity, DEPs primarily participated in biosynthetic processes, metabolic pathways, and the formation of ribosomes and proteasomes. Additionally, some proteins associated with senescence and programmed cell death were found to be upregulated in S-morph pistils, which may lead to the lower fruit set in S-morph flowers. Although the exact roles of these related proteins are not yet known, this was the first attempt to use an iTRAQ approach to analyze proteomes of heterostylous eggplant flowers, and these results will provide insights into biochemical events taking place during the development of heterostyly.