This study aimed to establish fabrication methods for two types of functional microcapsules (MCs) and to verify their fundamental performance in cementitious materials. The alkaline-supplying and CO2-absorbing MCs were evaluated independently. It was hypothesized that alkaline-supplying MCs could suppress carbonation while limiting strength loss to ≤20%, whereas CO2-absorbing MCs could increase long-term CO2 uptake by ≥30% relative to plain mortar. The alkaline-supplying MCs reduced the 24-week carbonation depth by approximately 35%, with a ~20% reduction in compressive strength at a 5% addition level. In contrast, CO2-absorbing MCs resulted in a ~30% strength reduction at 5% addition but increased CO2 uptake by 1.4 times that of plain mortar over 140 d. In conclusion, the results demonstrate that MCs can effectively impart carbonation-resistance or CO2-absorption functionality; however, a clear trade-off exists between functional enhancement and mechanical degradation due to capsule stiffness and density limitations. Because the two types of MCs rely on different mechanisms, they were tested in separate mortar systems. As this study focused on MC fabrication and fundamental functional verification, future work should optimize shell design, density control, and amine selection to balance structural performance with durability enhancement and CO2-sequestration capability.

Acquired resistance to 5-fluorouracil (5-FU) is a primary clinical challenge in colorectal cancer (CRC) treatment. Our study aimed to identify key factors predictive of 5-FU resistance and to elucidate their functional mechanisms by combining multi-omics analysis with experimental verification. The prognostic model was constructed based on the gene expression omnibus (GEO, GSE196900, GSE166555) and the cancer genome atlas (TCGA)-Colon Adenocarcinoma (COAD) datasets combined with regression analysis. Kaplan-Meier (K-M), receiver operating characteristic (ROC) curve, and nomogram were used to evaluate the predictive performance of the prognostic model. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) were used for functional enrichment analysis. Single-cell RNA sequencing (scRNA-seq, GSE166555) and qRT-PCR analysis were used to analyze the tumor microenvironment and gene expression. In cell experiments, CCK-8 assay measured IC50 value. Glycolysis metabolism was evaluated by detecting glucose consumption, lactic acid production, extracellular acidification rate (ECAR), and oxygen consumption rate (OCR); cell stemness was evaluated by sphere formation assay. A 5-gene prognostic model was successfully constructed, which could effectively distinguish the high-/low-risk groups of CRC patients and was significantly correlated with overall survival. Ribosome binding protein 1 (RRBP1) is highly expressed in cancer tissues of non-responders to chemotherapy. It is also highly expressed in tumor epithelial cells, and its high expression is closely related to aneuploidy characteristics, up-regulation of oncogenes, and activation of pro-survival pathways. Invitro experiments confirmed that knockdown of RRBP1 significantly enhanced the sensitivity of CRC cells to 5-FU and inhibited cell proliferation. Mechanistically, RRBP1 knockdown effectively reversed the enhanced glycolysis activity and stem cell-like properties of 5-FU-resistant cells. This study established RRBP1 as a key CRC prognostic factor and 5-FU resistance driver, operating through the regulation of cell glycolysis and stemness. RRBP1 emerges as a new biomarker and therapeutic target for predicting the efficacy of 5-FU.

Mining of key genes involved in the sulforaphane biosynthetic pathway of moringa and cloning, expression, and functional verification of MoMYR1.

Exercise-induced irisin attenuates ferroptosis in polycystic ovary syndrome by modulating the NCOA4-FTH pathway.

Tribolium castaneum CO2 controlled atmosphere stress screening and functional verification of CYP genes

The global prevalence of metabolic dysfunction-associated fatty liver disease (MAFLD) is increasing annually, significantly impairing patients' quality of life. Given the limitations of existing treatments, this study aims to investigate the effects of intermittent fasting (IF) on MAFLD and its underlying mechanisms. The liver tissues of four groups of mice were analyzed by bulk RNA sequencing: normal ad libitum diet (CD group), normal IF (iCD group), high-fat ad libitum diet (HFD group) and high-fat IF group (iHFD group). Differentially expressed genes (DEGs) were identified, followed by enrichment analyses including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA). Weighted gene co-expression network analysis (WGCNA) was used to identify related modules. The most highly correlated module genes were intersected with DEGs and analyzed by protein-protein interaction(PPI) network to identify key genes. The key genes were preliminarily verified by RT-qPCR. The function of the key gene was further verified by in vitro and in vivo experiments. IF significantly improved metabolic abnormalities and hepatic lipid deposition in MAFLD mice. A total of 331 DEGs were identified between the HFD and CD group, 379 DEGs between the iHFD and HFD group, and 142 DEGs were found to be common to both comparisons. Enrichment analysis showed that DEGs were mainly enriched in pathways related to fatty acid metabolism and inflammatory responses. WGCNA identified red and blue modules are most strongly correlated with MAFLD traits. After intersecting with DEGs, 32 genes were obtained. Based on PPI network analysis, we identified five key genes. After knocking down one of the key genes, Lrg1, in vitro and in vivo, we confirmed that Lrg1 may promote the expression of lipogenic genes such as Srebf1, Scd1, and Fasn via the PI3K-AKT pathway, thereby accelerating MAFLD progression. Transcriptome analysis elucidated the potential mechanism by which intermittent fasting improves MAFLD, highlighting the important role of fatty acid metabolism and inflammatory responses. Several key genes regulating MAFLD through IF were identified. Knocking down the key gene Lrg1 inhibited the expression of lipogenic genes and effectively slowed MAFLD progression.

Phytochelatin synthase (PCS) is crucial for synthesizing phytochelatins, cysteine-rich peptides vital for heavy metal detoxification in plants. Potato, a key staple crop in China, faces risks from soil heavy metal contamination, yet the genes involved in its detoxification, particularly PCS genes, remain underexplored. This study systematically identified and characterized the StPCS gene family in potato using genomic databases, uncovering five StPCS members distributed across three of the 12 potato chromosomes. Phylogenetic analysis classified StPCS proteins into three clades, while gene structure and motif analyses revealed high conservation in domain organization. Promoter region investigations identified stress-responsive elements in nearly all StPCS genes. Under cadmium (Cd) stress conditions, qPCR analysis indicated a significant upregulation of StPCS1 (5.73-fold) and StPCS2 (1.61-fold) transcript levels after 21 days compared to the control, whereas no obvious differences were observed at 7 days post-stress. Subsequent functional verification in yeast revealed that StPCS1 overexpression markedly improved Cd tolerance in transgenic yeast. In addition, analysis of cis-acting elements in the StPCS gene promoter combined with qPCR verification under MeJA and ABA stress conditions suggested that StPCS1 might be involved in Cd stress responses in potato through certain hormone signaling pathways. This study represents the first comprehensive analysis of the StPCS gene family in potato, clarifying its structural characteristics and characterizing the function of StPCS1 as a long-term Cd stress-responsive gene, which lays a solid foundation for investigating its role in heavy metal detoxification.

DENV virus (DENV) infection can cause various symptoms and organ damage, even severe dengue fever. However, the underlying host response products and interfering metabolic pathways and mechanisms of DENV infection remain unclear. In this study, we characterized the metabolites and metabolic pathway changes during DENV infection using liquid chromatography- (LC-MS) and gas chromatography-mass spectrometry (GC-MS). And identify the hub differentially expressed targets associated with major metabolism pathways combining transcriptomics. Plasma from adult patients infected with DENV infection was characterized by untargeted metabolomics using LC-MS and GC-MS. Potential diagnostic biomarkers for dengue fever were indicated using ROC curve analysis. KEGG and GSEA functional enrichment analysis was the strategy to determine the mechanisms of key metabolic pathways in dengue fever. Potential targets were identified by combining transcriptomic in Gene Expression Omnibus (GEO) datasets, and gene databases from GeneCards and the Comparative Toxicogenomics Database (CTD). A total of 41 dengue patients and 23 healthy volunteers were recruited for the study. 61 up-regulated and 136 down-regulated metabolites were identified via untargeted metabolomics. The top10 up-regulated metabolites with high AUC values included trans-cinnamic acid, L-Acetylcarnitine, SM(d17:1/17:0), 1,2,4,5-cyclohexanetetrol, 5-(hydroxymethyl) pyrrolidin-2-one, 1,2,3,4-tetrahydro-6-propanoylpyridine, 2-C-methyl-D-erythritol-4-phosphate, Physalolactone, S-Japonin, and 9-tridecynoic acid, and they were supposed to be the potential diagnostic biomarkers for dengue fever. The disturbance of ATP-binding cassette (ABC) transporters, protein digestion and absorption, aminoacyl-tRNA biosynthesis, mineral absorption, and D-amino acid metabolism were enriched in the metabolic pathways. ABCC5, ABCB1, and ABCG5 were identified as hub differentially expressed targets through transcriptome profiling and protein-protein interaction networks. The current study revealed a shift in metabolite profiles and disturbance in ABC transporters in dengue fever, which can be used for further functional verification.

We identified ABA- and drought-induced CtSnRK2.6 from Cynanchum thesioides; its protein localizes to the nucleus and plasma membrane. It boosts drought resistance of C. thesioides and Arabidopsis by regulating antioxidant and stress-responsive genes, while its silencing impairs C. thesioides' drought tolerance. CtSnRK2.6 interacts with CtPP2C1/3, and CtPP2C1 binds CtPYL1/5/6. This study clarifies its core role and interaction network in the ABA pathway, laying a theoretical foundation for elucidating C. thesioides' stress resistance mechanisms. Sucrose non-fermenting-1-related protein kinase 2 (SnRK2) is a core regulator in the plant abiotic stress response pathway. In this study, we identified a Cynanchum thesioides gene CtSnRK2.6 that is homologous to Arabidopsis AtSnRK2.6, and this gene is significantly induced by drought stress and abscisic acid (ABA) treatment. Subcellular localization analysis revealed that the CtSnRK2.6 protein is localized in the nucleus and plasma membrane. Functional verification results demonstrated that overexpression of CtSnRK2.6 significantly enhances osmotic stress tolerance in yeast and drought resistance in Arabidopsis; under drought conditions, the overexpressing plants not only maintain a superior phenotype, but also exhibit significantly increased fresh weight and chlorophyll content, markedly enhanced antioxidant enzyme activity, significantly reduced accumulation of reactive oxygen species (O₂⁻, H₂O₂) and malondialdehyde (MDA), as well as significantly upregulated expression levels of stress-responsive genes. Conversely, inhibition of CtSnRK2.6 expression via Tobacco Rattle Virus (TRV)-mediated gene silencing technology significantly impairs the drought resistance of C. thesioides. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays further confirmed that CtSnRK2.6 can directly interact with CtPP2C1 and CtPP2C3; CtPP2C1 can interact with CtPYL1, CtPYL5, and CtPYL6, respectively. Taken together, this study clarifies the key role of CtSnRK2.6 in enhancing drought resistance by regulating the plant antioxidant system, and reveals its interaction network with PYL and PP2C proteins, which are core components of the ABA signaling pathway. These findings provide important theoretical support for in-depth elucidation of the stress adaptation mechanism of C. thesioides.

The U6 promoter plays a pivotal role in the CRISPR/Cas9 system by driving the transcription of single guide RNA (sgRNA), which directs Cas9 to achieve precise genome editing. Endogenous U6 promoters typically exhibit superior transcriptional activation efficiency compared to exogenous counterparts, thereby enhancing the efficacy of genome editing. However, the endogenous U6 promoter in kenaf (Hibiscus cannabinusL.) remains uncharacterized. In this study, we conducted a homologous search of the kenaf genome using theArabidopsisU6 (AtU6-26) RNA sequence as a reference, identifying two candidate promoters, HcU6-1 and HcU6-14. Promoter fragments were amplified from the genomic DNA of kenaf cultivar 'Fuhong 952' and subsequently cloned into a GUS fusion expression vector. Histochemical staining revealed transcriptional activity for both promoters, with HcU6-14 demonstrating significantly stronger activity. To evaluate editing efficiency, we constructed a CRISPR/Cas9 vector containingHcALSsgRNA, driven by either the kenaf U6-14P promoter or the cotton U6-9P (GbU6-9P) promoter. Kenaf hairy roots were regenerated viaAgrobacteriumrhizogenes K599-mediated transformation. Sequencing analysis ofALSgene fragments from these hairy roots confirmed successful targeted editing when using the kenaf U6-14P promoter, whereas no base mutations were detected with the cotton U6 promoter. These findings highlight the superior editing efficiency of the kenaf U6 promoter and provide a critical foundation for advancing functional genomics research in kenaf.

Drought stress severely inhibits plant growth and yield, and plants have evolved various strategies to mitigate its effects. However, the genetic basis of photosynthetic traits and their responses during drought stress in Brassica napus (B. napus) remains poorly understood. In this study, we assessed photosynthetic traits in a natural population of 167 B. napus accessions under well-watered and mild drought stress conditions. Genome-wide association studies (GWAS) identified 106 quantitative trait locus (QTLs) associated with photosynthetic traits. Among these QTLs, a major QTL, qSC.A10.1, which associated with stomatal conductance under mild drought, was located. Within this region, a candidate gene, BnaA10.LEA4-5, which encodes a late embryogenesis abundant (LEA) protein, was identified. Functional verification revealed that BnaLEA4-5 promotes jasmonic acid (JA) biosynthesis, thereby reducing stomatal density and conductance and enhancing water use efficiency and drought resistance in B. napus. Further investigation showed that BnaLEA4-5 induces JA biosynthesis by upregulating AOS1 through the transcription factors EDT1 and RAP2.4, leading to MYC2-regulated reduction of stomatal density. These findings elucidate the genetic basis and molecular mechanism underlying photosynthetic adaptation to drought stress in B. napus and provide a genetic resource for genetic improvement of drought resistance in B. napus breeding.

Saussurea involucrata REVEILLE8, a circadian clock protein, confers salt tolerance by ROS signaling.

Multi-omics assessment of synthetic microbiome-mediated remediation of cyclotetramethylene tetranitroamine (HMX) contaminated water.

Identification of PI3K gene family in large yellow croaker (Larimichthys crocea): Expression patterns under Pseudomonas plecoglossicida infection and hypoxia stress, and functional verification of pik3r3b.

Identification of the DEAD-box gene family in apple (Malus domestica) and functional verification of MdRH28 under low-temperature stress.

Rhizospheric bacterial communities and growth-promoting bacteria functional verification in the reclaimed vineyards of Ningxia in Northwest China

Screening and identification of a novel PGAM5-specific inhibitor for attenuating multi-organ injury.

Efficient expression and functional verification of recombinant human type Ⅲ collagen in CHO cells.

Automotive System-on-Chips (SoCs) must meet stringent functional safety standards, such as ISO 26262 and IEC 61508, to ensure reliable operation under hardware faults. FPGA-based fault injection has emerged as a practical and cost-effective technique for functional safety verification. However, instrumentation-based methods face scalability challenges when applied to the high fault densities typical of automotive SoCs. To address these challenges, we propose a hybrid cascaded fault-injection controller architecture (HCCA-SAFE) that simultaneously reduces high-fanout global nets and eliminates long serial propagation paths. The architecture constrains enable-signal cluster width and distributes control across cascaded stages, improving timing results and routability under limited FPGA resources. The proposed architecture is evaluated on multiple open-source RISC-V processor cores. On openE902, HCCA-SAFE reduces net delay from 27.276 ns to 22.535 ns and achieves 32.2% and 63.8% lower net delay compared with the representative centralized and shift-chain approaches, respectively. On openE906, the proposed HCCA-SAFE limits the net delay to 12.959 ns and reduces the maximum control-signal fanout to 1763, respectively, compared with 25.825 ns and 40.442 ns in the conventional method. On openC906, the proposed design lowers the maximum control-signal fanout from 7725 to 570 and reduces the net delay to 7.506 ns. Furthermore, HCCA-SAFE produces results fully consistent with software-based RTL simulation, while delivering substantial performance gains. Speed-up factors of 127×, 206×, and 2123× are achieved on openE902, openE906, and openC906, respectively, with efficiency improvements scaling with processor complexity These results confirm that HCCA-SAFE delivers scalable, timing-robust fault-injection control suitable for large automotive SoCs.

High-Level Synthesis (HLS)-based VLSI design flows allow designers to start from high-level specifications in a general-purpose programming language like C/C++ and automatically generate an optimized hardware design in a hardware description language (HDL) like Verilog or VHDL. Supply of compromised computer-aided design (CAD) tools by an electronic design automation (EDA) vendor to the chip designers is a great threat that adversely affects the horizontal semiconductor business model. Recent works have examined the potential for security issues induced by a compromised HLS CAD tool and demonstrated how HLS is a prime candidate for hardware Trojan (HT) insertion into any underlying design since it is hard to correlate the high-level description to the generated register-transfer level (RTL) code. Further, it has been shown that the use of compiler-generated intermediate representation (IR) as a likely attack vector for inserting HT in the RTL during an HLS-based IC design flow, taking advantage of the lack of automated methods to analyse logic inside a complex LLVM IR. In this work, we propose, implement, and evaluate a novel HLS security verification framework by leveraging the modern large language models (LLMs). Specifically, we focus on detecting the HTs introduced using Black-Hat HLS and HLS-IRT toolchain by performing functional verification using LLM. The experimental results show that LLMs have an impressive ability to analyse and automatically identify these hardware security anomalies.