Mutant p53 increases exosome-mediated transfer of mi R-21-3p and mi R-769-3p to promote pulmonary metastasis

Objective: Tumor metastasis is a complex, multistep process that depends on tumor cells and their communication with the tumor microenvironment. A p53 gain-of-function mutant has been shown to enhance the tumorigenesis, invasion, and metastasis abilities of tumor cells. This study aimed to investigate the roles of p53 R273 H mutation in the tumor microenvironment.Methods: The in vitro and in vivo effects of the p53 R273 H mutant on the invasion and metastasis of HCT116 cells were investigated. Exosomes from wild-type and HCT116-TP53(R273 H) cells were cocultured with mouse embryonic fibroblasts(MEFs). The roles of differentially expressed exosomal micro RNAs identified by microarray analysis were investigated. The functions of the p53 R273 H mutant in tumor cells were also investigated via gene expression microarray and quantitative polymerase chain reaction(q PCR) analyses.Results: Introducing p53 R273 H mutant into HCT116 cells significantly potentiated pulmonary metastasis in vivo. In the presence of exosomes derived from HCT116-TP53(R273 H) cells, the exosomes were taken up by MEFs and became activated. Microarray analysis showed that the p53 R273 H mutation increased the exosomal levels of mi R-21-3 p and mi R-769-3 p. Intriguingly, in clinical samples, mi R-21-3 p and mi R-769-3 p levels were significantly higher in patients with a p53 mutation than in those without this mutation. Furthermore, both mi R-21-3 p and mi R-769-3 p activated fibroblasts and exerted a synergistic effect via their target genes on the transforming growth factor-β(TGF-β)/Smad signaling pathway. The activated fibroblasts excreted cytokine TGF-β and may have reciprocally induced cancer cells to undergo epithelial-mesenchymal transition(EMT). Indeed, HCT116-TP53(R273 H) cells showed increased expression of ZEB1 and SNAI2 and decreased transcription of several cell adhesion molecules.Conclusions: The mutant p53-exosomal mi R-21-3 p/mi R-769-3 p-fibroblast-cytokine circuit appears to be responsible for communication between tumor and stromal cells, with exosomal mi RNAs acting as a bridge. mi R-21-3 p and mi R-769-3 p are potential predictive markers of pulmonary metastasis and candidate targets for therapeutic interventions.

Bioinspired engineering ADSC nanovesicles thermosensitive hydrogel enhance autophagy of dermal papilla cells for androgenetic alopecia treatment

Androgenic alopecia(AGA)is a highly prevalent form of non-scarring alopecia but lacks effective treatments.Stem cell exosomes have similar repair effects to stem cells,suffer from the drawbacks of high cost and low yield yet.Cell-derived nanovesicles acquired through mechanical extrusion exhibit favorable biomimetic properties similar to exosomes,enabling them to efficiently encapsulate substantial quantities of therapeutic proteins.In this study,we observed that JAM-A,an adhesion protein,resulted in a significantly increased the adhesion and resilience of dermal papilla cells to form snap structures against damage caused by dihydrotestosterone and macrophages,thereby facilitating the process of hair regrowth in cases of AGA.Consequently,adipose-derived stem cells were modified to overexpress JAM-A to produce engineered JAM-A overexpressing nanovesicles(JAM-A^(OE)@NV).The incorporation of JAM-A^(OE)@NV into a thermosensitive hydrogel matrix(JAM-A^(OE)@NV Gel)to effectively addresses the limitations associated with the short half-life of JAM-A^(OE)@NV,and resulted in the achievement of a sustained-release profile for JAM-A^(OE)@NV.The physicochemical characteristics of the JAM-A^(OE)@NV Gel were analyzed and assessed for its efficacy in promoting hair regrowth in vivo and vitro.The JAM-A^(OE)@NV Gel,thus,presents a novel therapeutic approach and theoretical framework for promoting the treatment of low cell adhesion diseases similar to AGA.

Redundant CAMTA Transcription Factors Negatively Regulate the Biosynthesis of Salicylic Acid and N-Hydroxypipecolic Acid by Modulating the Expression of SARD1 and CBP60g

Two signal molecules, salicylic acid (SA) and N-hydroxypipecolic acid (NHP), play critical roles in plant immunity. The biosynthetic genes of both compounds are positively regulated by master immune-regulating transcription factors SARD1 and CBP60g. However, the relationship between the SA and NHP pathways is unclear. CALMODULIN-BINDING TRANSCRIPTION FACTOR 1 (CAMTA1), CAMTA2, and CAMTA3 are known redundant negative regulators of plant immunity, but the underlying mechanism also remains largely unknown. In this study, through chromatin immunoprecipitation and electrophoretic mobility shift assays, we uncovered that CBP60g is a direct target of CAMTA3, which also negatively regulates the expression of SARD1, presumably via an indirect effect. The autoimmunity of camta3-1 is suppressed by sard1 cbp60g double mutant as well as ald1 and fmo1, two single mutants defective in NHP biosynthesis. Interestingly, a suppressor screen conducted in the camta1/ 2/ 3 triple mutant background yielded various mutants blocking biosynthesis or signaling of either SA or NHP, leading to nearly complete suppression of the extreme autoimmunity of camta1/ 2/ 3, suggesting that the SA and NHP pathways can mutually amplify each other. Together, these results reveal that CAMTAs repress the biosynthesis of SA and NHP by modulating the expression of SARD1 and CBP60g, and that the SA and NHP pathways are coordinated to optimize plant immune response.

Transformer-based ensemble deep learning model for EEG-based emotion recognition

Emotion recognition is one of the most important research directions in the field of brain–computer interface(BCI).However,to conduct electroencephalogram(EEG)-based emotion recognition,there exist difficulties regarding EEG signal processing;moreover,the performance of classification models in this regard is restricted.To counter these issues,the 2022 World Robot Contest successfully held an affective BCI competition,thus promoting the innovation of EEG-based emotion recognition.In this paper,we propose the Transformer-based ensemble(TBEM)deep learning model.TBEM comprises two models:a pure convolutional neural network(CNN)model and a cascaded CNN-Transformer hybrid model.The proposed model won the abovementioned affective BCI competition’s final championship in the 2022 World Robot Contest,demonstrating the effectiveness of the proposed TBEM deep learning model for EEG-based emotion recognition.

Ammonia volatilization and availability of Cu,Zn induced by applications of urea with and without coating in soils

Ammonia volatilization and the distribution of Cu and Zn were investigated in two types of soil treated with coated and uncoated urea.The rate of ammonia volatilization in two weeks after fertilizing with coated urea was 8% in soil 1 （soil derived from river alluvial deposits in Dongting Lake Plain） and 5.15% in soil 2 （red soil derived from quaternary red clay）,about half the rates observed when fertilizing with common urea,implying that the hydrolysis speed of the coated urea was lower than for common urea,and that the coated urea can increase nitrogen use efficacy.As for the availability of Cu and Zn,their concentrations decreased in the first week after fertilization,and then increased,which was contrary to the effect of treatment on soil pH.For example,when the pH was 7.99,there was 0.79mg/kg exchangeable Cu and 0.85mg/kg exchangeable Zn in the soil derived from river alluvial deposits in Dongting Lake Plain.However,the concentrations of exchangeable Cu and Zn were generally lower for the common urea treatments than those with the coated urea because the peak pH for the common urea treatment was greater.The concentrations of these elements correlated well with pH in the range 4–8 in second order polynomial fits.

Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A-driven angiogenesis

Background:Non-healing wounds are an intractable problem of major clinical relevance.Evidence has shown that dermal papilla cells(DPCs)may regulate the wound-healing process by secreting extracellular vesicles(EVs).However,low isolation efficiency and restricted cell viability hinder the applications of DPC-EVs in wound healing.In this study,we aimed to develop novel 3D-DPC spheroids(tdDPCs)based on self-feeder 3D culture and to evaluate the roles of tdDPC-EVs in stimulating angiogenesis and skin wound healing.Methods:To address the current limitations of DPC-EVs,we previously developed a self-feeder 3D culture method to construct tdDPCs.DPCs and tdDPCs were identified using immunofluorescence staining and flow cytometry.Subsequently,we extracted EVs from the cells and compared the effects of DPC-EVs and tdDPC-EVs on human umbilical vein endothelial cells(HUVECs)in vitro using immunofluorescence staining,a scratch-wound assay and a Transwell assay.We simultaneously established a murine model of full-thickness skin injury and evaluated the effects of DPC-EVs and tdDPC-EVs on wound-healing efficiency in vivo using laser Doppler,as well as hematoxylin and eosin,Masson,CD31 andα-SMA staining.To elucidate the underlying mechanism,we conducted RNA sequencing(RNA-seq)of tdDPC-EV-and phosphate-buffered saline-treated HUVECs.To validate the RNA-seq data,we constructed knockdown and overexpression vectors of Krüppel-like factor 4(KLF4).Western blotting,a scratch-wound assay,a Transwell assay and a tubule-formation test were performed to detect the protein expression,cell migration and lumen-formation ability of KLF4 and vascular endothelial growth factor A(VEGFA)in HUVECs incubated with tdDPC-EVs after KLF4 knockdown or overexpression.Dual-luciferase reporter gene assays were conducted to verify the activation effect of KLF4 on VEGFA.Results:We successfully cultured tdDPCs and extracted EVs from DPCs and tdDPCs.The tdDPC-EVs significantly promoted the proliferation,lumen formation and migration of HUVECs.Unlike DPC-EVs,tdDPC-EVs exhibited significant advantages in terms of promoting angiogenesis,accelerating wound healing and enhancing wound-healing efficiency both in vitro and in vivo.Bioinformatics analysis and further functional experiments verified that the tdDPC-EV-regulated KLF4/VEGFA axis is pivotal in accelerating wound healing.Conclusions:3D cultivation can be utilized as an innovative optimization strategy to effectively develop DPC-derived EVs for the treatment of skin wounds.tdDPC-EVs significantly enhance wound healing via KLF4/VEGFA-driven angiogenesis.

A 3D Homogenized Model for Nonlinear Wave Interaction with Randomly Distributed Microcracks

This paper presents a 3D homogenized model able to simulate the nonlinear effects generated by the interaction of ultrasonic waves with microcracks.A hexahedral element with one horizontal elliptical crack is constructed as a reference model.The reference model is smeared to be orthotropic but with different moduli in tension and compression to account for stiffness asymmetry due to crack opening and closure.Different from the existing homogenized models that usually simplify the representative volume element as a homogeneous part and require only one constitutive model for the equivalent material of the whole structure,we assign the constitutive relationship of the same reference model to all the finite elements but with random principal material orientations to take randomly oriented microcracks into consideration.In this way,the randomness of distributed microcracks can be considered in the framework of continuum mechanics.In this manner,the experimentally observed nonlinear effects,such as the generation of both even and odd harmonics,can be reproduced.The developed model has been verified for a 3D bar and a 3D plate.Particularly,the influence of crack density on the amplitude of higher harmonics is analyzed.

Theoretical simulation of performances in CIGS thin-film solar cells with cadmiumfree buffer layer

Copper indium gallium selenium （CIGS） thin film solar cells have become one of the hottest topics in solar energy due to their high photoelectric transformation efficiency. To real applications, CIGS thin film is covered by the buffer layer and absorption layer. Traditionally, cadmium sulfide （CdS） is inserted into the middle of the window layer （ZnO） and absorption layer （CIGS） as a buffer layer. However, the application of the GIGS/CdS thin film solar cells has been limited because of the environmental pollution resulting from the toxic cadmium atom. Although zinc sulfide （ZnS） has been proposed to be one of the candidates, the performance of such battery cells has not been investigated. Here, in this paper, we systematically study the possibility of using zinc sulfide （ZnS） as a buffer layer. By including the effects of thickness, concentration of a buffer layer, intrinsic layer and the absorbing layer, we find that photoelectric transformation efficiency of ZnO/ZnS（n）/CIGS（i）/CIGS（p） solar cell is about 17.22%, which is qualified as a commercial solar cell. Moreover, we also find that the open-circuit voltage is -0.60 V, the short-circuit current is -36.99 mA/cm2 and the filled factor is -77.44%. Therefore, our results suggest that zinc sulfide may be the potential candidate of CdS as a buffer layer.

Correction to:A 3D Homogenized Model for Nonlinear Wave Interaction with Randomly Distributed Microcracks

Correction to:Acta Mechanica Solida Sinica https://doi.org/10.1007/s10338-022-00347-y In this article the list of references has been wrongly formatted.It should have shown as below.The original article has been corrected.