Gram-positive bacterial membrane vesicles (MVs) are natural biomaterials with great potential in intercellular communication and antipathogen defense, while the thick cell wall leads to extremely low natural yields, severely limiting their large-scale production and practical application. Herein, we developed a green and efficient physical strategy using cold atmospheric plasma (CAP) to induce high-yield MVs from Bacillus subtilis (B. subtilis), a widely used probiotic and biocontrol agent. CAP treatment achieved a 10.19-fold increase in MVs particle yield over the untreated control, outperforming traditional ultrasonic disruption. The synergistic action of CAP-generated acidic stress, RONS, and physical discharge caused cell wall/membrane damage and intracellular oxidative stress, activating the SOS response, endolysin pathway, and iron metabolism regulation as candidate pathways associated with MVs release. Cytotoxicity and plant safety assays confirmed the biosafety of MVs. CAP-induced MVs (CAP MVs) retained a conserved spherical nanostructure with uniform size distribution, and shared 99.96% protein components with control MVs, ensuring biosafety. Meanwhile, CAP MVs were enriched with functional proteins related to oxidative stress response and cell wall degradation. Both control and CAP MVs exhibited antipathogen activity against Gram-positive/negative bacteria and fungi, with CAP MVs showing significantly enhanced efficacy against Pseudomonas syringae and Fusarium graminearum. Notably, the B. subtilis MVs was first identified as a key inducer in antipathogen activity. This study provided a green strategy for high-yield MVs production and uncovered a novel application for B. subtilis MVs, advancing the biogenesis of extracellular vesicles in prokaryotes.

Poly(3-hydroxybutyrate) (P3HB) is a promising sustainable plastic alternative, valued for its inherent biodegradability, biocompatibility, and desirable material properties. While structural modification of P3HB has largely centered on side-chain functionalization, its end-group functionalization remains relatively unexplored. Herein, we demonstrate that modulation with epoxides in the trimetallic-catalyzed ring-opening polymerization of beta-butyrolactone has enabled the synthesis of various end-functionalized P3HBs. In the trimetallic aluminum system, epoxides have switched the initiating group from chloride to alkoxide, thus transforming the propagating species from carboxylate to alkoxide. In contrast, the trimetallic chromium system has exhibited a three-fold enhancement in activity (TOF up to 800 h-1) and five-fold rise in molecular weight (Mn = 47.6 kg mol-1) upon epoxide addition, while maintaining a narrow dispersity (& Dstrok; similar to 1.26). Mechanistic insight reveals that epoxides converted the chloride to an alkoxide initiator, while the propagation still proceeded via a robust carboxylate species with configurational inversion at the methine carbon atom. This strategy offers a new route to prepare high-molecular-weight P3HB products with desired end-functionalization.

The dense melanin–protein nanoparticles (MPNs) of cuttlefish ink constrain conventional hot-water (WE), ultrasound-assisted (UAE), and alkali–acid cycling (ABE) extractions to a trade-off between bioactive release and flavor preservation. Here, Sepia pharaonis ink was processed by coupling a choline chloride–glycerol deep eutectic solvent (ChCl:Gly, 1:2 mol/mol) with Alcalase hydrolysis (DES-assisted enzymatic processing, EE) and benchmarked against WE, UAE, and ABE. EE exhibited a distinctively elevated profile of umami amino acids and flavor nucleotides, yielding the highest equivalent umami concentration. GC-O-MS and GC-IMS collectively resolved 92 and 58 volatiles; aroma extract dilution analysis (AEDA) resolved 30 key odorants with flavor dilution factor (FD) ≥ 27. Docking and 100 ns molecular dynamics gave binding energies of −6.2 to −8.68 kcal mol−1 for five odorants against four olfactory receptors, stabilized by hydrogen bonding and hydrophobic contacts. Olfactory EEG showed the highest power spectral density (20.21 μV2 Hz−1) and strongest α/β-band activation for EE. 16S rRNA sequencing, short-chain fatty acid (SCFA) quantification, and cytokine profiling further showed that EE delivered the highest α-diversity, largest SCFA yield, and lowest pro-inflammatory response, establishing it as the optimal route. In a DSS-induced chronic ulcerative colitis with depression-like behavior (CUC) model, EE was associated with improved tight junction expression, shifted gut microbiota composition, and lower LPS translocation, lowered inflammatory cytokines, attenuated cerebral oxidative stress and glial activation, preserved hippocampal neurons, and ameliorated depressive and cognitive phenotypes. Spearman correlations revealed a microbiota–gut–brain axis cascade of microbiota compositional shifts, barrier repair, attenuated LPS translocation, and reduced neuroinflammation, establishing a transferable chemical-engineering paradigm for valorizing cephalopod MPN by-products.

Chemical looping ammonia synthesis offers a promising alternative to the Haber-Bosch process, yet the synthesis of effective nitrogen carriers from stable oxides remains a central challenge. A plasma-driven chemical looping strategy is developed, in which MgO nanoparticles are plasma-nitrided to generate Mg-based nitrogen carriers in a gas-powder hybrid rotating gliding arc plasma. Effective plasma-solid coupling enables concurrent activation of N2 and MgO, promoting MgO nitridation under non-equilibrium conditions. In situ optical emission spectroscopy and high-speed imaging reveal a transition from breakdown-gliding to arc-gliding discharge, together with intensified vibrational excitation and characteristic Mg emission, indicating plasma-enabled MgO activation and subsequent nitridation. Introducing 2% Ar further promotes reactive nitrogen formation, likely through enhanced excitation/ionization and energy-transfer processes in the N2/Ar plasma. Under the same applied voltage and a fixed total gas flow rate of 10 L min-1 , the NH3 synthesis rate increased from 334.1 mu mol & sdot;g- 1 & sdot;h- 1 in pure N2 to 401.1 mu mol & sdot;g- 1 & sdot;h- 1 with 2% Ar, corresponding to an approximately 20% improvement. Combined plasma diagnostics, product characterization, and DFT calculations suggest that surface/near-surface MgO nitridation mainly proceeds through a nitrogen-assisted oxygen-abstraction pathway, with vacancy-mediated nitrogen incorporation likely contributing under localized high-energy interfacial conditions. This work establishes a renewable-electricity-driven route for Mg-based nitrogen carrier synthesis and provides mechanistic insight into plasma-enabled chemical looping ammonia synthesis.

Plant bacterial diseases caused by Gram-negative pathogens pose severe threats to global agricultural production and food security, while traditional chemical pesticides suffer from growing antimicrobial resistance and environmental pollution. RNA interference (RNAi) is ineffective in prokaryotes, creating a critical technical gap for green bacterial disease control. Here, we developed a novel precision antimicrobial platform by integrating cetyltrimethylammonium bromide-modified zeolitic imidazolate framework-8 (CTAB@ZIF-8, cZIF-8) with the CRISPR-Cas12a system, enabling electroporation-free, competent-cell-independent delivery of CRISPR genetic drugs into phytopathogenic bacteria. Through comparative genomics across 27 species, we identified ftsZ as a core conserved essential target with exceptional bactericidal potency. The cZIF-8 nanocarrier achieved exceptional plasmid delivery efficiencies of ∼78% in E. coli and ∼ 40% in P. aeruginosa, overcoming the outer membrane barrier of Gram-negative bacteria. Upon activation within the cell, Cas12a mediates sequence-specific cleavage of the ftsZ gene, leading to bacterial cell death. Tomato fruit infection models demonstrated that this cZIF-8/CRISPR system significantly attenuated soft rot symptoms, reduced lesion expansion and fruit weight loss, without impairing fruit quality, thereby demonstrating its potential for field-relevant control of phytopathogenic infections. This work establishes a CRISPR-based pesticide strategy for the sustainable and eco-friendly management of plant pathogens.

The asymmetric alternating copolymerization of meso-epoxide and cyclic anhydrides provides an efficient access to enantiopure polyesters.Contrary to the extensive investigation of the stereochemistry resulting from epoxide building block,the chirality from anhydride and the configurational match with epoxide remain elusive.Herein,we discover that the bimetallic chromium catalysts have led to an obvious en-hancement in terms of reactivity and enantioselectivity for the asymmetric copolymerization of meso-epoxide with various non-symmetric chiral anhydrides.Up to 97％ee was obtained during the asymmetric copolymerization of cyclohexene oxide(CHO)with(R)-methylsuccinic anhydride(R-MSA),and three-or four-carbon chiral centers were simultaneously installed in the aliphatic polyester backbone.In particular,the different combinations of stereochemistry in epoxide and anhydride building blocks considerably affect the thermal properties and crystalline behaviors of the resulting polyesters.This study uncovers an interesting method for regulating polymer crystallinity via matching the chirality of different monomers.

Ovarian cancer (OC), as a malignant tumor, currently lacks effective screening early diagnosis measures. Clinical biomarkers CA-125 and HE4 are limited by false positives and insufficient sensitivity. Therefore, it's of great significance to search for new biomarker and construct sensitive detection method. We found a novel circRNA biomarker (hsa_circ_0049101) by RNA sequencing, and simultaneously propose a strategy, which integrates reverse transcription rolling circle amplification (RT-RCA) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas12a to amplify and detect novel circRNA biomarker. This strategy use Dual Cas12a protein (FnCas12a and LbCas12a) and Multiplex CrRNA (DCMC-CRISPR) to enhance detection sensitivity. The sensitivity mechanism of CRISPR to detect circRNA was verified in detail. The DCMC-CRISPR assay exhibited a broad detection range of 2000 pM to 0.5 fM and the limit of detection (LOD) as low as 0.5 fM. The DCMC-CRISPR system has 4–11 times higher sensitivity than single-crRNA CRISPR/Cas12a system. Clinical assessment of RNA extracts from patient’s peripheral blood of 22 clinical OC patients and 28 controls demonstrates the DCMC-CRISPR strategy outperformed CA-125, HE4, and the ROMA index. The assay demonstrated comparable performance to RT-qPCR, exhibiting favorable sensitivity and specificity in this pilot cohort. The DCMC-CRISPR platform offers a promising solution for circRNA biomarker screening and circRNA diagnostic. It highlights the possibility of expanding its applicability to address other cancer diseases.

Plant pathogens pose significant threats to agricultural productivity and economic stability; therefore, the development of green broad-spectrum fungicides is critical to the long-term sustainability of agriculture. Here, zeolitic imidazolate framework nanoparticles (ZIF-8 NPs) with different charge properties were synthesized by a one-pot method and modified by different surfactants under the condition of a water phase at room temperature, which had an excellent bactericidal effect on model Gram-negative bacteria (Escherichia coli) and plant pathogenic bacteria (Xanthomonas axonopodis pv. citri and Pseudomonas syringae). Notably, the IC90 values of CTAB@ZIF-8 NPs for these bacterial strains were all below 50 mu g/mL. In addition, ZIF-8 NPs also had antibacterial effects on fungi (Magnaporthe grisea) and oomycetes (Phytophthora infestans). ZIF-8 NPs can destroy the cell wall and cell membrane, eliminate the biofilm, produce ROS, cause DNA damage, and lead to cell death. Together, this work may provide ideas for the development of a promising, sustainable, efficient, broad-spectrum agricultural antimicrobial agent.

Polyhydroxyalkanoates (PHAs) have emerged as compelling and sustainable alternatives to conventional polymers, distinguished by their inherent biocompatibility and enzymatic degradability. Nevertheless, the chemosynthetic PHAs face constraints in terms of functional versatility and compromised property tunability. Here, we report a bimetallic ligand platform enabling the yttrium-catalyzed stereoselective ring-opening polymerization of various beta-lactones, yielding functionalized PHAs with high activity (TOF up to 21,600 h-1) and syndiotacticity (P r up to 0.94). Mechanistic insights reveal that the intramolecular cooperation and exchange between the yttrium centers with heterochirality eliminate the less active species during chain propagation. In particular, the syndio-enriched poly(4-(phenoxymethyl)-2-propiolactone) overcomes the inherent brittleness of the corresponding defect-free PHAs, while imparting competitive permeability, mechanical and robust adhesion properties compared to commodity polymers, and superior optical transparency to the prospective biobased plastics.

Verticillium wilt,caused by the infamous pathogen Verticillium dahliae,presents a primary constraint on cotton cul-tivation worldwide.The complexity of disease resistance in cotton and the largely unexplored interaction dynamics between the cotton plant host and V.dahliae pathogen pose a crucial predicament for effectively managing cotton Verticillium wilt.Nevertheless,the most cost-effective approach to controlling this disease involves breeding and cul-tivating resistant cotton varieties,demanding a meticulous analysis of the mechanisms underlying cotton's resistance to Verticillium wilt and the identification of pivotal genes.These aspects constitute focal points in disease-resistance breeding programs.In this review,we comprehensively discuss genetic inheritance associated with Verticillium wilt resistance in cotton,the advancements in molecular markers for disease resistance,the functional investiga-tion of resistance genes in cotton,the analysis of pathogenicity genes in V.dahliae,as well as the intricate interplay between cotton and this fungus.Moreover,we delve into the future prospects of cutting-edge research on cotton Verticillium wilt,aiming to proffer valuable insights for the effective management of this devastating fungus.

Polyhydroxyalkanoates (PHAs) have served as promising alternatives to traditional petroleum-based plastics. Chemical synthesis of stereoregular PHAs via stereoselective copolymerization of racemic-epoxides with carbon monoxide (CO) has not yet been achieved. Here, the design of trimetallic ligand platform featuring various electronic nature and steric demand enables CrIII-catalyzed stereoselective copolymerization of racemic-epoxides with CO, yielding 9 types of functionalized PHA products having 0.70 syndiotacticity. Kinetic study has revealed that trimetallic complex favored the intramolecular chain propagation for the preparation of high molecular weight PHAs with high reactivity and moderate syndiotacticity, while monometallic complex promoted the intermolecular chain propagation toward isotactic-enriched PHAs.

This work successfully developed a new green and low-cost preparation method of zeolitic imidazolate framework-8 (ZIF-8) by adding sodium hydroxide as a proton removing agent and surfactant to reduce the amount of 2-methylimidazole and the size of ZIF-8. Using surfactants cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulfate (SDS) to modify ZIF-8. The modified ZIF-8 showed selective adsorption behaviors for Rhodamine B (Rh B), methylene blue (MB), methyl orange (MO), and acid yellow 36 (AC36). ZIF-8 modified by SDS was more likely to adsorb MB and Rh B cationic dyes than ZIF-8 modified by CTAB, with removal rates of 85.3 and 90.1%, respectively. The removal rates of anionic MO and AC36 by CTAB-modified ZIF-8 were 98.7 and 80.4%, respectively. Because of the selectivity of modified ZIF-8, it can separate specific dyes from mixed dyes. Fourier transform infrared spectrometry and zeta potential analysis showed that the adsorption of dyes by modified ZIF-8 was carried out by electrostatic interaction, π–π stacking and hydrogen bonding.

The biological functions of circular RNA (circRNAs) in cancers have garnered significant attention, particularly for their potential as biomarkers. However, the roles of circRNAs in ovarian cancer (OC) and their applicability for early detection of this malignancy remain underexplored. We performed RNA sequencing on ovarian cancer cell lines to identify circRNAs associated with OC. The functional mechanisms of the identified circRNAs were elucidated through bioinformatics analysis. The discriminating ability of biomarkers was assessed using receiver operating characteristic (ROC) analysis. RNA sequencing analysis revealed that 170 known circRNAs were correlated with ovarian cancer. Through the circRNA-miRNA-mRNA regulatory network, we identified 9 circRNAs that interact with 8 miRNAs, subsequently regulating the expression of 324 mRNAs. Functional enrichment analysis, protein-protein interaction (PPI) network analysis, and hub gene analysis indicated that these circRNAs and miRNAs may play a role in regulating MAPK, Wnt, and ErbB signaling pathways. We validated these circRNAs and miRNAs expression profiles in cell, tissue, and plasma samples, identifying four candidates-hsa_circ_0049101, hsa_circ_0007440, hsa_circ_0006935, and hsa-miR-338-3p-that expression level positively correlate with ovarian cancer development. These markers were then combined into a circRNA and miRNA detection (CMD) panel for ovarian cancer detection. The area under the curve (AUC) values obtained from ROC analysis demonstrated that these individual candidates, as well as the CMD panel, exhibited superior discriminatory ability for OC compared to traditional biomarkers such as CA125, HE4, and the ROMA index in our sample set, which included 28 healthy controls and 22 ovarian cancer patients. Notably, the CMD panel showed exceptional potential for distinguishing early-stage OC samples from healthy controls, achieving an AUC of 1. In this study, we elucidated the functional mechanisms of a set of circRNAs associated with OC through multi-omics analysis and demonstrated that the combination of circRNAs and miRNAs into a biomarker panel holds significant potential for early detection of ovarian cancer.

The aberrant expression of RNAs in ovarian cancer (OC) progression highlights their potential as clinical biomarkers. However, rapid and accurate quantification of these RNAs in biosamples remains a significant challenge. In this study, we develop a modular isothermal rolling circle amplification (RCA)-activated Cas12a loop-enhanced (MIRACLE) amplification method for circRNA and miRNA quantification without the need of reverse transcription. In this design, isothermal amplification of modular DNA can be initiated by target-specific RCA primers or miRNAs, with the amplification products subsequently recognized by the Cas12a system to generate measurable signals. When integrated with a multi-volume sliding chip (SlipChip) platform, this MIRACLE method enables portable, rapid and ultra-sensitive quantification of these two types of RNA. Under optimized conditions, this platform exhibits detection limits of 0.125 copies per μL for circRNA and 0.326 copies per μL for miRNA, covering a 5-log dynamic range from 10-1 to 103 copies per μL within 35 min. The platform was validated using OC cell lines and clinical blood samples. It successfully profiled OC RNA biomarkers (hsa_circ_0049101 and hsa-miR-338-3p) and effectively distinguished between early and advanced stages of OC. These results show a strong correlation with RT-qPCR (R2 = 0.953 for circRNA and R2 = 0.947 for miRNA). This work establishes a versatile CRISPR-microfluidic platform for cancer diagnosis. Its modular design allows for adaptation to detect other cancer-related RNA biomarkers, thereby addressing critical needs in precision oncology.

The plasma-activated water (PAW) containing numerous reactive species can facilitate chitin degradation. Given the intricate interplay between PAW treatment and the diverse activities of chitinolytic enzymes, further investigation is imperative for enhancing the chitin bioconversion efficiency. This study revealed that PAW-treated chitin exhibited improved degradability toward BtLPMO10A, endochitinases OfChtI, OfChtII-B4C1, and exochitinase OfChi-h. Furthermore, H2O2 in PAW boosted BtLPMO10A, whereas the soluble constituents in PAW generated during chitin pretreatment inhibited OfChi-h. Notably, this inhibition effect toward OfChi-h can be mitigated by the addition of β-N-acetylhexosaminidase. In the end, the synergy among the chitinolytic enzyme was also promoted by PAW pretreatment. On this as a basis, a chitin degradation strategy using a combination of PAW treatment and an enzyme cocktail was applied to degrade chitin, achieving a chitin conversion yield of 97% within 2 h. This strategy could also be applied to the degradation of other polysaccharides, such as cellulose.

Single-modality cancer therapies face limitations from tumor heterogeneity, drug resistance, and immune evasion. Combination therapies, particularly those that merge nucleic acid-based and immune-based therapeutics, offer promise by leveraging synergy; however, both face delivery challenges. This study reports a liquid-liquid phase separation (LLPS)-driven therapeutic coacervate system designed to simultaneously deliver nucleic acid drugs and immunostimulators for synergistic antitumor treatment. Short elastin-like polypeptide-antisense oligonucleotide (E-ASO) conjugates are synthesized to form membraneless coacervates via LLPS. These E-ASO coacervates directly internalize into cells and potently silence anti-apoptotic gene, Bcl-2, across six distinct cancer cell lines. By incorporating the stimulator of interferon gene (STING) agonist cyclic GMP-AMP (cGAMP), E-ASO-cGAMP coacervates are generated, which simultaneously downregulate Bcl-2 and activate the STING pathway, triggering cytokine production and immune activation. In melanoma and hypopharyngeal carcinoma mouse models, E-ASO-cGAMP coacervates demonstrate significantly enhanced antitumor efficacy compared to monotherapies, with potent and consistent tumor suppression across both immunologically distinct tumor types. The platform shows no observable systemic toxicity in vital organs. This LLPS-driven coacervate system establishes a new model for combined anticancer therapy, overcoming delivery barriers to synergistically integrate gene silencing and immune activation and provide a translatable strategy for solid tumor treatment.

Tumor heterogeneity is associated with poor prognosis and drug resistance, leading to therapeutic failure. Here, we used tumor evolution analysis to determine the intra- and intertumoral heterogeneity of high-grade serous ovarian cancer (HGSOC) and analyze the correlation between tumor heterogeneity and prognosis, as well as chemotherapy response, through single-cell and spatial transcriptomic analysis. We collected and curated 28 HGSOC patients’ single-cell transcriptomic data from five datasets. Then, we developed a novel text-mining-based machine-learning approach to deconstruct the evolutionary patterns of tumor cell functions. We then identified key tumor-related genes within different evolutionary branches, characterized the microenvironmental cell compositions that various functional tumor cells depend on, and analyzed the intra- and intertumoral heterogeneity as well as the tumor microenvironments. These analyses were conducted in relation to the prognosis and chemotherapy response in HGSOC patients. We validated our findings in two spatial and seven bulk transcriptomic datasets (total: 1,030 patients). Using transcriptomic clusters as proxies for functional clonality, we identified a significant increase in tumor cell state heterogeneity that was strongly correlated with patient prognosis and treatment response. Furthermore, increased intra- and intertumoral functional clonality was associated with the characteristics of cancer-associated fibroblasts (CAFs). The spatial proximity between CXCL12-positive CAFs and tumor cells, mediated through the CXCL12/CXCR4 interaction, was highly positively correlated with poor prognosis and chemotherapy resistance in HGSOC. Finally, we constructed a panel of 24 genes through statistical modeling that correlate with CXCL12-positive fibroblasts and can predict both prognosis and the response to chemotherapy in HGSOC patients. Our study offers insights into the collective behavior of tumor cell communities in HGSOC, as well as potential drivers of tumor evolution in response to therapy. There was a strong association between CXCL12-positive fibroblasts and tumor progression, as well as treatment outcomes.

Fiber quality is a major breeding goal in cotton, but phenotypically direct selection is often hindered. In this study, we identified fiber quality and yield related loci using GWAS based on 2.97 million SNPs obtained from 10.65× resequencing data of 1,081 accessions. The results showed that 585 novel fiber loci, including two novel stable SNP peaks associated with fiber length on chromosomes At12 and Dt05 and one novel genome regions linked with fiber strength on chromosome Dt12 were identified. Furthermore, by means of gene expression analysis, GhM_A12G0090 and GhM_D05G1692, involved in fiber length and one gene, GhM_D12G3135, associated with fiber strength were identified. Additionally, 14 consistent and stable superior haplotypes were identified, and 25 accessions were detected as possessing these 14 superior haplotype in breeding. This study provides fundamental insight relevant to identification of genes associated with fiber quality and yield will enhance future efforts toward improvement of upland cotton.