FREE1 regulates phagophore closure in plants.

Adenosine monophosphate-activated protein kinase (AMPK), the central energy sensor in more complex eukaryotes, can activate macroautophagy/autophagy upon cellular energy deficiency. However, the regulatory role of nutrient sensing in mediating phagophore closure to generate an autophagosome remains unknown. The evolutionarily conserved endosomal sorting complexes required for transport (ESCRT) machinery has been postulated to regulate phagophore sealing, yet the signaling pathway modulating the ESCRT complex relocation from multivesicular body (MVB) to phagophore for closure remains unknown. We recently identified a plant unique pleiotropic protein FREE1 (FYVE DOMAIN PROTEIN REQUIRED FOR ENDOSOMAL SORTING 1), which is phosphorylated by the plant energy sensor SnRK1 (SNF1-related kinase 1) and bridges the ATG conjugation system and ESCRT machinery to regulate phagophore sealing upon nutrient starvation. This study elucidated the bona fide roles and underlying mechanism of cellular energy-sensing pathways in regulating compartment sealing.

Comparative peptidomics analysis in the discovery of umami peptides from Chinese Douchi.

Douchi is a kind of traditional Chinese fermented soybean product with outstanding umami taste. Besides the umami amino acids in Douchi, peptides were also considered as an important contributor for the umami taste of Douchi. Peptides with molecular weight below 0.66 kDa accounted for more than 50 % in all samples except for TongChuan Douchi, and a total of 421 peptides were identified from the ten kinds of Douchi samples by using LC-MS/MS. Combined with sensory evaluation results, 19 peptides containing Glu, Asp or known umami peptide sequences were chosen as potential umami peptides via PLS-DA and RDA analysis. Among them, 17 soluble peptides exhibited obvious umami taste and the threshold of 7 peptides were lower than MSG solution. Especially, the VD was detected with a minimum umami taste threshold at 0.16 mg/mL. The results indicated that the umami peptides might be the important components affecting the umami taste of Douchi.

Frequency-domain terahertz optoacoustics for non-contact quantitative detection of gas, liquid, and solid samples.

Terahertz optoacoustics (THz-OA) combines the advantages of abundant molecular characteristic absorptions in a terahertz band and the low attenuation through ultrasonic detection. Frequency-domain THz-OA, benefiting from the compact and the low cost of a continuous-wave THz source, has been used in gas detection and sensing. However, liquid and solid detections are hard to achieve due to the sensitivity limitation of existing technologies. Here we present a high-sensitivity frequency-domain THz-OA system with customized optoacoustic cells to accomplish non-contact quantitative detection of gas, liquid, and solid samples. The relationships between signal amplitudes and sample concentration, volume and temperature are discussed separately, revealing a potential application of this technology.

Circular RNA circTMEM59 inhibits progression of pancreatic ductal adenocarcinoma by targeting miR-147b/SOCS1: An in vitro study.

Purpose: This study aimed to detect the role and mechanism of circTMEM59 in pancreatic ductal adenocarcinoma (PDAC). Methods: 66 paired PDAC tissues and normal samples were harvested from patients diagnosed and undergoing pancreatic cancer surgery in our hospital. The expression of circTMEM59 in PDAC tissues and cell lines was detected. Based on bioinformatics information, the circTMEM59 mimics, miR-147b mimics, miR-147b inhibitor and si-suppressor of cytokine signaling 1 (SOCS1) were transfected into PDAC cells. The expression levels of circTMEM59, miR-147b and SOCS1 were detected by quantitative real-time polymerase chain reaction (qRT-PCR). RNA interaction was confirmed by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Cell invasion and proliferation were evaluated by Transwell and Cell Counting Kit-8 (CCK-8) assays. The protein expression was detected by Western blot. Results: CircTMEM59 was confirmed to be downregulated in PDAC tumor tissues and cells. Low expression of circTMEM59 was closely correlated with the short survival time and poor clinicopathological characteristics. By up-regulating the expression of circTMEM59 in PDAC cells, cell proliferation, invasion and epithelial-mesenchymal transition (EMT) were inhibited. More importantly, miR-147b could be sponged by circTMEM59, and knockdown of miR-147b inhibited progression of PDAC cells. Further study revealed that SOCS1 was targeted by miR-147b. SOCS1 expression was negatively related to miR-147b expression and positively related to circTMEM59 expression in PDAC tissues. Upregulated miR-147b and downregulated SOCS1 could rescue the effects of circTMEM59 on cell proliferation, EMT and invasion. Conclusion: Our data indicated that circTMEM59 inhibited cell proliferation, invasion and EMT of PDAC by regulating miR-147b/SOCS1 axis.

A chemometric-assisted method for automatic, rapid and non-targeted detection of multi-pesticides in plant-derived foods by gas chromatography-mass spectrometry.

An automatic, rapid and non-targeted detection method for multi-pesticides in plant-derived foods was developed by gas chromatography-mass spectrometry and chemometrics. In this method, a novel algorithm named moving window iterative target transformation factor analysis was proposed. Although there are challenges of peak overlapping and background interference, the retention time and corrected mass spectra of unknown pesticides can be automatically obtained through iteration calculation in the 'moving window' with reference to the pesticide mass spectral library. One mixed pesticide standard and nine varieties of plant-derived foods were investigated with the proposed method. By contrast, a fast temperature programme was used to shorten detection time compared to the standard temperature programme. For the mixed standard, the mass spectra and retention times of all 39 pesticides were successfully obtained from the overlapping signal. Furthermore, all spiked pesticides were successfully detected in plant-derived foods within 10 min using a fast temperature programme.

Introducing Specific Iodine Ions in Perovskite-Based Nanocomplex to Cater for Versatile Biomedical Imaging and Tumor Radiotherapy.

Multimodal biomedical imaging and imaging-guided therapy have garnered extensive attention owing to the aid of nanoagents with the aim of further improving the therapeutic efficacy of diseases. The ability to engineer nanocomplexes (NCs) or control how they behave within an organism remains largely elusive. Here, a multifunctional nanoplatform is developed based on stabilized I-doped perovskite, CsPbBr3 -x Ix @SiO2 @Lip-c(RGD)2 (PSL-c(RGD)2 ) NCs. In particular, by regulating the amount of regular I- ions introduced, the fluorescence emission spectrum of perovskite-based NCs can be modulated well to match the requirement for biomedical optical imaging at the scale from molecule, cell to mouse; doping 125 I enables the nanoformulation to be competent for single-photon emission computed tomography (SPECT) imaging; the introduction of 131 I- imparts the NCs with the capability for radiotherapy. Through facile manipulation of specific iodine ions, this nanoplatform exhibits a remarkable ability to match multifunctional biomedical imaging and tumor therapy. In addition, their in vivo behavior can be manipulated by adjusting the thickness of the silica shell and the surface polarity for more practical applications. These experimental explorations offer a novel approach for engineering desirable multimodal NCs to simultaneously image and combat malignant tumors.

An improved multi-scale gradient generative adversarial network for enhancing classification of colorectal cancer histological images.

Introduction: Deep learning-based solutions for histological image classification have gained attention in recent years due to their potential for objective evaluation of histological images. However, these methods often require a large number of expert annotations, which are both time-consuming and labor-intensive to obtain. Several scholars have proposed generative models to augment labeled data, but these often result in label uncertainty due to incomplete learning of the data distribution. Methods: To alleviate these issues, a method called InceptionV3-SMSG-GAN has been proposed to enhance classification performance by generating high-quality images. Specifically, images synthesized by Multi-Scale Gradients Generative Adversarial Network (MSG-GAN) are selectively added to the training set through a selection mechanism utilizing a trained model to choose generated images with higher class probabilities. The selection mechanism filters the synthetic images that contain ambiguous category information, thus alleviating label uncertainty. Results: Experimental results show that compared with the baseline method which uses InceptionV3, the proposed method can significantly improve the performance of pathological image classification from 86.87% to 89.54% for overall accuracy. Additionally, the quality of generated images is evaluated quantitatively using various commonly used evaluation metrics. Discussion: The proposed InceptionV3-SMSG-GAN method exhibited good classification ability, where histological image could be divided into nine categories. Future work could focus on further refining the image generation and selection processes to optimize classification performance.

Exploring the Release of Elastin Peptides Generated from Enzymatic Hydrolysis of Bovine Elastin via Peptide Mapping.

To enhance the understanding of enzymatic hydrolysis and to accelerate the discovery of key bioactive peptides within enzymatic products, this research focused on elastin as the substrate and investigated the variations in peptide profiles and the production of key bioactive peptides (those exceeding 5% of the total) and their impacts on the biological activity of the hydrolysates. Through the application of advanced analytical techniques, such as stop-flow two-dimensional liquid chromatography and ultra-high-performance liquid chromatography-tandem mass spectrometry, the research tracks the release and profiles of peptides within elastin hydrolysates (EHs). Despite uniform peptide compositions, significant disparities in peptide concentrations were detected across the hydrolysates, hinting at varying levels of bioactive efficacy. A comprehensive identification process pinpointed 403 peptides within the EHs, with 18 peptides surpassing 5% in theoretical maximum content, signaling their crucial role in the hydrolysate's bioactivity. Of particular interest, certain peptides containing sequences of alanine, valine, and glycine were released in higher quantities, suggesting Alcalase® 2.4L's preference for these residues. The analysis not only confirms the peptides' dose-responsive elastase inhibitory potential but also underscores the nuanced interplay between peptide content, biological function, and their collective synergy. The study sets the stage for future research aimed at refining enzymatic treatments to fully exploit the bioactive properties of elastin.

The seven rice vacuolar sorting receptors localize to prevacuolar compartments.

Vacuolar sorting is critically important in plants as it regulates the mobilization of proteins and plays a major role in important agricultural traits like yield and seed protein content. Vacuolar sorting receptors (VSRs) are integral membrane proteins that mediate protein trafficking from the Golgi apparatus to the vacuole via the intermediate membrane-bound prevacuolar compartment (PVC)/multivesicular body (MVB). VSR proteins, such as an 80 kD (BP-80) from pea, also serve as markers for PVC/MVB. Dissecting VSR-mediated protein trafficking pathways may provide ways to enhance agronomic traits and crop yield. Green fluorescence protein (GFP) fusions with the seven Arabidopsis (Arabidopsis thaliana) VSRs were previously shown to localize to PVCs in transgenic tobacco BY-2 cells. The Rice (Oryza sativa) genome contains seven VSRs (OsVSR1-7), but little is known about their subcellular localizations. Here we studied the subcellular localization of OsVSR1-7 b y using a reporter approach, in which GFP-OsVSR1-7 fusions containing the transmembrane domain (TMD) and cytoplasmic tail (CT) of individual OsVSR were expressed in the protoplasts of rice, transgenic tobacco BY-2 cells and transgenic rice plants. Immunofluorescent labelling studies and confocal laser scanning microscope observation demonstrated that the seven OsVSRs are localized to PVCs and form ring-like structures upon wortmannin treatment. Therefore, we have verified the subcellular localization of OsVSR1-7 in this study. The OsVSRs tagged with GFP can serve as PVCs/MVBs markers in rice for the future studies.

Effect of bevacizumab administered prior to transarterial chemoembolization on the therapeutic effects of lenvatinib given post-TACE in primary liver cancer patients.

OBJECTIVE: To determine the therapeutic effects of lenvatinib combined with bevacizumab following transarterial chemoembolization (TACE) in patients with primary liver cancer. MATERIALS AND METHODS: 100 patients with primary liver cancer were recruited in the period from January 2020 to January 2021 and allocated with randomization into a control group (n = 50) and a test (bevacizumab) group (n = 50). The patients in the control group received lenvatinib for 4 weeks following TACE, whereas those in the test group received bevacizumab for 6 weeks prior to TACE and subsequent therapy with lenvatinib for 4 weeks. The serum concentration of interferon-γ (INF-γ), interleukin-10 (IL-10), soluble interleukin-2 receptor (sIL-2R), interleukin-12 (IL-12), superoxide dismutase (SOD), total antioxidant capacity (TAOC), glutathione (GSH), malondialdehyde (MDA), total bilirubin (TBil), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), carbohydrate antigen 242 (CA242), CA724, α-fetoprotein (AFP) and carcinoembryonic antigen (CEA) were determined in both groups at the commencement of treatment in January 2020 and 12 months later and compared with the observed therapeutic effects. RESULTS: The concentrations of CA242, CEA, CA724, and AFP in the bevacizumab group were lower than those in the control group (p < 0.05). The concentration of IL-12 and INF-γ in the bevacizumab group were higher, but the levels of IL-10 and sIL-2R lower than in the control group (p < 0.05). In the bevacizumab group, the level of MDA was lower, whereas the levels of TAOC, SOD, and GSH were higher than those in the control group (p < 0.05). The bevacizumab group also had lower levels of ALT, TBil, and AST and a higher level of ALP than control group (p < 0.05). The response rate based on tumor status (size, progression) in the bevacizumab group was higher than in the control group (p < 0.05). CONCLUSION: The therapeutic effects of lenvatinib following TACE in primary liver cancer are significantly greater when combined with bevacizumab administered for 6 weeks prior to TACE.

An autophagy-inducing stapled peptide induces mitochondria dysfunction and triggers autotic cell death in triple-negative breast cancer.

Autophagy is a lysosome-dependent bulk degradation process essential for cell viability but excessive autophagy leads to a unique form of cell death termed autosis. Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer with notable defect in its autophagy process. In previous studies, we developed stapled peptides that specifically targeted the essential autophagy protein Beclin 1 to induce autophagy and promote endolysosomal trafficking. Here we show that one lead peptide Tat-SP4 induced mild increase of autophagy in TNBC cells but showed potent anti-proliferative effect that could not be rescued by inhibitors of programmed cell death pathways. The cell death induced by Tat-SP4 showed typical features of autosis including sustained adherence to the substrate surface, rupture of plasma membrane and effective rescue by digoxin, a cardioglycoside that blocks the Na+/K+ ATPase. Tat-SP4 also induced prominent mitochondria dysfunction including loss of mitochondria membrane potential, elevated mitochondria reactive oxygen species and reduced oxidative phosphorylation. The anti-proliferative effect of Tat-SP4 was confirmed in a TNBC xenograft model. Our study uncovers three notable aspects of autosis. Firstly, autosis can be triggered by moderate increase in autophagy if such increase exceeds the endogenous capacity of the host cells. Secondly, mitochondria may play an essential role in autosis with dysregulated autophagy leading to mitochondria dysfunction to trigger autosis. Lastly, intrinsic autophagy deficiency and quiescent mitochondria bioenergetic profile likely render TNBC cells particularly susceptible to autosis. Our designed peptides like Tat-SP4 may serve as potential therapeutic candidates against TNBC by targeting this vulnerability.

Electronic Structure Regulation and Surface Reconstruction of Iron Diselenide for Enhanced Oxygen Evolution Activity.

Regulating the electronic structure of active sites and monitoring the evolution of the active component is essential to improve the intrinsic activity of catalysts for electrochemical reactions. Herein, a highly efficient pre-electrocatalyst of iron diselenide with rich Se vacancies achieved by phosphorus doping (denoted as P-FeSe2 ) for oxygen evolution reaction (OER) is reported. Systematically experimental and theoretical results show that the formed Se vacancies with phosphorus doping can synergistically modulate the electronic structure of FeSe2 and facilitate OER kinetics with the resulting enhanced electrical conductivity and electrochemical surface area. Importantly, the in situ formed FeOOH species on the surface of the P-FeSe2 nanorods (denoted as P-FeOOH(Se)) during the OER process acts as an active component to efficiently catalyze OER and exhibits a low overpotential of 217 mV to reach 10 mA cm-2 with good durability. Promisingly, an alkaline electrolyzer assembled with P-FeOOH(Se) and Pt/C electrodes requires an ultra-low cell voltage of 1.50 V at 10 mA cm-2 for overall water splitting, which is superior to the RuO2 || Pt/C counterpart and most of the state-of-the-art electrolyzers, demonstrating the high potential of the fabricated electrocatalyst by P doping strategy to explore more highly efficient selenide-based catalysts for various reactions.

Photo-Responsive Phase-Separating Fluorescent Molecules for Intracellular Protein Delivery.

Cellular membranes, including the plasma and endosome membranes, are barriers to outside proteins. Various vehicles have been devised to deliver proteins across the plasma membrane, but in many cases, the payload gets trapped in the endosome. Here we designed a photo-responsive phase-separating fluorescent molecule (PPFM) with a molecular weight of 666.8 daltons. The PPFM compound condensates as fluorescent droplets in the aqueous solution by liquid-liquid phase separation (LLPS), which disintegrate upon photoirradiation with a 405 nm light-emitting diode (LED) lamp within 20 min or a 405 nm laser within 3 min. The PPFM coacervates recruit a wide range of peptides and proteins and deliver them into mammalian cells. Photolysis disperses the payload from condensates into the cytosolic space. Altogether, a type of small molecules that are photo-responsive and phase separating are discovered; their coacervates can serve as transmembrane vehicles for intracellular delivery of proteins, whereas photo illumination triggers the cytosolic distribution of the payload.

Spatiotemporal organization and correlation of tip-focused exocytosis and endocytosis in regulating pollen tube tip growth.

Pollen tube polar growth is a key cellular process during plant fertilization and is regulated by tip-focused exocytosis and endocytosis. However, the spatiotemporal dynamics and localizations of apical exocytosis and endocytosis in the tip region are still a matter of debate. Here, we use a refined spinning-disk confocal microscope coupled with fluorescence recovery after photobleaching for sustained live imaging and quantitative analysis of rapid vesicular activities in growing pollen tube tips. We traced and analyzed the occurrence site of exocytic plasma membrane-targeting of Arabidopsis secretory carrier membrane protein 4 and its subsequent endocytosis in tobacco pollen tube tips. We demonstrated that the pollen tube apex is the site for both vesicle polar exocytic fusion and endocytosis to take place. In addition, we disrupted either tip-focused exocytosis or endocytosis and found that their dynamic activities are closely correlated with one another basing on the spatial organization of actin fringe. Collectively, our findings attempt to propose a new exocytosis and endocytosis-coordinated yin-yang working model underlying the apical membrane organization and dynamics during pollen tube tip growth.

Effect of High Efficiency Digestion and Utilization of Organic Iron Made by Saccharomyces cerevisiae on Antioxidation and Caecum Microflora in Weaned Piglets.

Organic iron is expected to replace inorganic iron used in diets as an iron source. Organic iron possesses high absorption efficiency and low fecal iron excretion. This study aims to study the effect of organic iron produced by Saccharomyces cerevisiae (yeast iron) on digestion, utilization, antioxidation and caecum microflora in weaned piglets. In total, 20 piglets that had been weaned after 28 days were divided into 4 groups, each of which followed a different basal diet. The basal diet of each of these 4 groups contained, respectively, 104 mg/kg iron (ferrous sulfate, CON), 84 mg/kg iron (yeast iron, LSC), 104 mg/kg iron (yeast iron, MSC) or 124 mg/kg iron (yeast iron, HSC). This experiment lasted 35 d. The apparent digestibility of iron in LSC, MSC and HMS was higher than that in CON (p < 0.01) and the fecal iron content in LSC, MSC and HMS was lower than that in CON (p < 0.01). Serum iron contents in LSC, MSC and HMS were higher than that in CON (p < 0.01). The iron contents of the heart, lungs, liver, kidney and left gluteus muscle in the MSC and HMS groups were higher than that in CON and LSC (p < 0.05). Serum catalase, glutathione peroxidase, superoxide dismutase activity, superoxide anion, glutathione, hydroxyl free radical scavenging rate, total antioxidant capacity, and liver superoxide anion clearance rate and peroxidase in MSC and HMS were higher than that in CON and LSC (p < 0.05). The contents of nitric oxide and peroxide of the weaned piglets in MSC and HMS were lower than that in CON and LSC (p < 0.05). The abundance of Firmicutes, Blautia and Peptococcus in LSC, HSC and MSC was higher than that in CON (p < 0.01). The abundance of Lactobacillus in CON and LSC was higher than that in MSC and HSC (p < 0.01). The abundance of Acinetobacter, Streptococcus and Prevotella in LSC, MSC and HSC was lower than that in CON (p < 0.01). The results suggested that a diet containing 84 mg/kg iron of yeast iron has the same effect as a diet containing 104 mg/kg iron of ferric sulfate, and that a diet containing 104 or 124 mg/kg iron of yeast iron is superior to a diet containing 104 mg/kg iron of ferric sulfate.

Computer-aided detection and prognosis of colorectal cancer on whole slide images using dual resolution deep learning.

PURPOSE: Rapid diagnosis and risk stratification can provide timely treatment for colorectal cancer (CRC) patients. Deep learning (DL) is not only used to identify tumor regions in histopathological images, but also applied to predict survival and achieve risk stratification. Whereas, most of methods dependent on regions of interest annotated by pathologist and ignore the global information in the image. METHODS: A dual resolution DL network based on weakly supervised learning (WDRNet) was proposed for CRC identification and prognosis. The proposed method was trained and validated on the dataset from The Cancer Genome Atlas (TCGA) and tested on the external dataset from Affiliated Cancer Hospital and Institute of Guangzhou Medical University (ACHIGMU). RESULTS: In identification task, WDRNet accurately identified tumor images with an accuracy of 0.977 in slide level and 0.953 in patch level. Besides, in prognosis task, WDRNet showed an excellent prediction performance in both datasets with the concordance index (C-index) of 0.716 ± 0.037 and 0.598 ± 0.024 respectively. Moreover, the results of risk stratification were statistically significant in univariate analysis (p < 0.001, HR = 7.892 in TCGA-CRC, and p = 0.009, HR = 1.718 in ACHIGMU) and multivariate analysis (p < 0.001, HR = 5.914 in TCGA-CRC, and p = 0.025, HR = 1.674 in ACHIGMU). CONCLUSIONS: We developed a weakly supervised resolution DL network to achieve precise identification and prognosis of CRC patients, which will assist doctors in diagnosis on histopathological images and stratify patients to select appropriate therapeutic schedule.

Dual resolution deep learning network with self-attention mechanism for classification and localisation of colorectal cancer in histopathological images.

AIMS: Microscopic examination is a basic diagnostic technology for colorectal cancer (CRC), but it is very laborious. We developed a dual resolution deep learning network with self-attention mechanism (DRSANet) which combines context and details for CRC binary classification and localisation in whole slide images (WSIs), and as a computer-aided diagnosis (CAD) to improve the sensitivity and specificity of doctors' diagnosis. METHODS: Representative regions of interest (ROI) of each tissue type were manually delineated in WSIs by pathologists. Based on the same coordinates of centre position, patches were extracted at different magnification levels from the ROI. Specifically, patches from low magnification level contain contextual information, while from high magnification level provide important details. A dual-inputs network was designed to learn context and details simultaneously, and self-attention mechanism was used to selectively learn different positions in the images to enhance the performance. RESULTS: In classification task, DRSANet outperformed the benchmark networks which only depended on the high magnification patches on two test set. Furthermore, in localisation task, DRSANet demonstrated a better localisation capability of tumour area in WSI with less areas of misidentification. CONCLUSIONS: We compared DRSANet with benchmark networks which only use the patches from high magnification level. Experimental results reveal that the performance of DRSANet is better than the benchmark networks. Both context and details should be considered in deep learning method.

Gas Chromatography-Mass Spectrometry Metabolite Analysis Combined with Transcriptomic and Proteomic Provide New Insights into Revealing Cuticle Formation during Pepper Development.

The cuticle plays an important role for the quality of pepper fruit. However, the molecular mechanism of cuticle formation in pepper fruit remains unclear. Our results showed that the wax was continuously accumulated during pepper development, while the cutin monomer first increased and then decreased. Hexadecanoic acid and 10,16-hydroxyhexadecanoic acid were the main components of wax and cutin, respectively. Combined with transcriptome and proteome, the formation patterns of wax and cutin polyester network for pepper cuticle was proposed. The 18 pairs of consistent expression genes and proteins involved in cuticle formation were revealed. Meanwhile, 12 key genes were screened from fatty acid biosynthesis, biosynthesis of unsaturated fatty acids, fatty acid elongation, cutin, suberine, and wax biosynthesis, glycerolipid metabolism, and transport pathway. This study would provide important candidate genes and theoretical basis for the molecular mechanism of cuticle formation, which is essential for the breeding of peppers.

TM9SF4 is an F-actin disassembly factor that promotes tumor progression and metastasis.

F-actin dynamics is crucial for many fundamental properties of cancer cells, from cell-substrate adhesion to migration, invasion and metastasis. However, the regulatory mechanisms of actin dynamics are still incompletely understood. In this study, we demonstrate the function of a protein named TM9SF4 in regulating actin dynamics and controlling cancer cell motility and metastasis. We show that an N-terminal fragment (NTF) cleaved from TM9SF4 can directly bind to F-actin to induce actin oxidation at Cys374, consequently enhancing cofilin-mediated F-actin disassembly. Knockdown of TM9SF4 reduces cell migration and invasion in ovarian cancer cells A2780, SKOV3 and several high grade serous ovarian cancer lines (HGSOCs). In vivo, knockdown of TM9SF4 completely abolishes the tumor growth and metastasis in athymic nude mice. These data provide mechanistic insights into TM9SF4-mediated regulation of actin dynamics in ovarian cancer cells.

TRPM2 Promotes Atherosclerotic Progression in a Mouse Model of Atherosclerosis.

Atherosclerosis is a chronic inflammatory arterial disease characterized by build-up of atheromatous plaque, which narrows the lumen of arteries. Hypercholesterolemia and excessive oxidative stress in arterial walls are among the main causative factors of atherosclerosis. Transient receptor potential channel M2 (TRPM2) is a Ca2+-permeable cation channel activated by oxidative stress. However, the role of TRPM2 in atherosclerosis in animal models is not well studied. In the present study, with the use of adeno-associated virus (AAV)-PCSK9 and TRPM2 knockout (TRPM2-/-) mice, we determined the role of TRPM2 in hypercholesterolemia-induced atherosclerosis. Our results demonstrated that TRPM2 knockout reduced atherosclerotic plaque area in analysis of En face Oil Red O staining of both whole aortas and aortic-root thin sections. Furthermore, TRPM2 knockout reduced the expression of CD68, α-SMA, and PCNA in the plaque region, suggesting a role of TRPM2 in promoting macrophage infiltration and smooth-muscle cell migration into the lesion area. Moreover, TRPM2 knockout reduced the expression of ICAM-1, MCP-1, and TNFα and decreased the ROS level in the plaque region, suggesting a role of TRPM2 in enhancing monocyte adhesion and promoting vascular inflammation. In bone-marrow-derived macrophages and primary cultured arterial endothelial cells, TRPM2 knockout reduced the production of inflammatory cytokines/factors and decreased ROS production. In addition, a TRPM2 antagonist N-(p-amylcinnamoyl) anthranilic acid (ACA) was able to inhibit atherosclerotic development in an ApoE-/- mouse model of atherosclerosis. Taken together, the findings of our study demonstrated that TRPM2 contributes to the progression of hypercholesterolemia-induced atherosclerosis. Mechanistically, TRPM2 channels may provide an essential link that can connect ROS to Ca2+ and inflammation, consequently promoting atherosclerotic progression.