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Abstract Neovaginas are surgically constructed to correct uterovaginal agenesis in women with Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome or as part of gender-affirming surgery for transfeminine individuals. Understanding the assembly of the neovaginal microbiota is crucial for guiding its management. To address this, we conducted a longitudinal study on MRKH patients following laparoscopic peritoneal vaginoplasty. Our findings reveal that the early microbial assemblage exhibited stochastic characteristics, accompanied with a notable bloom of Enterococcus faecalis and genital Mycoplasmas. While both the pre-surgery dimple microbiota and the fecal microbiota constituted the primary species pool, the neovaginal microbiota developed into a microbiota that resembled that of a normal vagina at 6–12 months post-surgery, albeit with a bacterial vaginosis (BV)-like structure. By 2–4 years post-surgery, the neovaginal microbiota had further evolved into a structure closely resembling with the homeostatic pre-surgery dimple microbiota. This concords with the development of the squamous epithelium in the neovagina and highlights the pivotal roles of progressive selective forces imposed by the evolving neovaginal environment and the colonization tropism of vaginal species. Notably, we observed that strains of Lactobacillus crispatus colonizing the neovagina primarily originated from the dimple. Since L. crispatus is generally associated with vaginal health, this finding suggests potential avenues for future research to promote its colonization.

Abstract The incidence of young-onset colorectal cancer (yCRC) has been increasing in recent decades, but little is known about the gut microbiome of these patients. Most studies have focused on old-onset CRC (oCRC), and it remains unclear whether CRC signatures derived from old patients are valid in young patients. To address this, we assembled the largest yCRC gut metagenomes to date from two independent cohorts and found that the CRC microbiome had limited association with age across adulthood. Differential analysis revealed that well-known CRC-associated taxa, such as Clostridium symbiosum, Peptostreptococcus stomatis, Parvimonas micra and Hungatella hathewayi were significantly enriched (false discovery rate

Cadmium (Cd) is an environmental endocrine disruptor. Despite increasing research about the metabolic effects of Cd on HepG2 cells, information about the metabolic effects of Cd on insulin resistance HepG2 (IR-HepG2) cells is limited. Currently, most individuals with diabetes are exposed to Cd due to pollution. Previously, we reported that Cd exposure resulted in decreased blood glucose levels in diabetic mice, the underlying mechanism deserves further study. Therefore, we used palmitic acid (0.25 mM) to treat HepG2 cells to establish IR-HepG2 model. IR-HepG2 cells were exposed to CdCl2 (1 μM and 2 μM). Commercial kits were used to measure glucose production, glucose consumption, ROS and mitochondrial membrane potential. Western blot and qRT-PCR were used to measure the proteins and genes of glucose metabolism. In the current study setting, we found no significant changes in glucose metabolism in Cd-exposed HepG2 cells, but Cd enhanced glucose uptake, inhibited gluconeogenesis and activated the insulin signaling pathway in IR-HepG2 cells. Meanwhile, we observed that Cd caused oxidative stress and increased the intracellular calcium concentration and inhibited mitochondrial membrane potential in IR-HepG2 cells. Cd compensatingly increased glycolysis in IR-HepG2 cells. Collectively, we found Cd ameliorated glucose metabolism disorders in IR-HepG2 cells. Furthermore, Cd exacerbated mitochondrial damage and compensatory increased glycolysis in IR-HepG2 cells. These findings will provide novel insights for Cd exposure in insulin resistant individuals.

ABSTRACT Beneficial interactions between plants and rhizosphere fungi can enhance plant adaptability during drought stress. However, harnessing these interactions will require an in-depth understanding of the response of fungal community assembly to drought. Herein, by using different varieties of wheat plants, we analyzed the drought-induced changes in fungal community assembly in rhizosphere and bulk soil. We demonstrated that drought significantly altered the fungal communities, with the contribution of species richness to community beta diversity increased in both rhizosphere and bulk soil compartments during drought stress. The stochastic processes dominated fungal community assembly, but the relative importance of deterministic processes, mainly homogeneous selection, increased in the drought-stressed rhizosphere. Drought induced an increase in the relative abundance of generalists in the rhizosphere, as opposed to specialists, and the top 10 abundant taxa that enriched under drought conditions were predominantly generalists. Notably, the most abundant drought-enriched taxon in rhizosphere was a generalist, and the corresponding Chaetomium strain was found capable of improving root length and activating ABA signaling in wheat plants through culture-based experiment. Together, these findings provide evidence that host plants exert a strong influence on rhizospheric fungal community assembly during stress and suggest the fungal communities that have experienced drought have the potential to confer fitness advantages to the host plants.IMPORTANCEWe have presented a framework to integrate the shifts in community assembly processes with plant-soil feedback during drought stress. We found that environmental filtering and host plant selection exert influence on the rhizospheric fungal community assembly, and the re-assembled community has great potential to alleviate plant drought stress. Our study proposes that future research should incorporate ecology with plant, microbiome, and molecular approaches to effectively harness the rhizospheric microbiome for enhancing the resilience of crop production to drought.

Abstract Genotyping‐by‐sequencing (GBS) is a widely used strategy for obtaining large numbers of genetic markers in model and non‐model organisms. In crop plants, GBS‐derived marker datasets are frequently used to perform quantitative trait locus (QTL) mapping. In some plant species, however, high heterozygosity and complex genome structure mean that researchers must use care in handling GBS data to conduct QTL mapping most effectively. Such outbred crops include most of the perennial grass and tree species used for bioenergy. To identify strategies for increasing accuracy and precision of QTL mapping using GBS data in outbred crops, we conducted an empirical study of SNP‐calling and genetic map‐building pipeline parameters in a Miscanthus sinensis population, and a complementary simulation study to estimate the relationship between genome‐wide error rate, read depth, and marker number. The bioenergy grass Miscanthus is an obligate outcrossing species with a recent (diploidized) whole‐genome duplication. For the study of empirical M. sinensis data, we compared two SNP‐calling methods (one non‐reference‐based and one reference‐based), a series of depth filters (12×, 20×, 30×, and 40×) and two map‐construction methods (i.e., marker ordering: linkage‐only and order‐corrected based on a reference genome). We found that correcting the order of markers on a linkage map by using a high‐quality reference genome improved QTL precision (shorter confidence intervals). For typical GBS datasets of between 1000 and 5000 markers to build a genetic map for biparental populations, a depth filter set at 30× to 40× applied to outbred populations provided a genome‐wide genotype‐calling error rate of less than 1%, improved accuracy of QTL point estimates and minimized type I errors for identifying QTL. Based on these results, we recommend using a reference genome to correct the marker order of genetic maps and a robust genotype depth filter to improve QTL mapping for outbred crops.

The issue of ocean corrosion in coastal areas, particularly in concrete structures partially buried in the soil has attracted wide attention has garnered attention due to the unique challenges it presents. This study investigates the enhanced chloride ions intrusion in concrete due to bidirectional unsaturated gradients between the parts buried in soil and those exposed to the air. A 90-day chloride salt erosion experiment was conducted on both fully buried and semi-buried concrete across five different soil environments to analyze the transport properties of chloride ions. Drilling powder sampling and the detection of free chloride ion content in the concrete specimens were performed at intervals of 30, 60, and 90 days. The results indicate that the chloride ions in semi-buried concrete exhibit bidirectional unsaturated migration toward the air-exposed end, leading to significantly peak chloride ion concentrations at the air-exposed end compared to those subjected to wet-dry cycles in a pure salt solution. Specifically, after a 90-day cycle, the total chloride contents at the air-exposed end of semi-buried concrete in pure sandy soil, 50 % clay soil, and pure clay soil increased by 68.8 %, 43.1 %, and 16.4 %, respectively. Soils with lower permeability intensified the vertical unsaturation gradient within the concrete, accelerating chloride ion migration towards the exposed end and resulting in higher accumulation at elevated positions. The research work underscores the critical impact of bi-directional unsaturated transport on concrete durability in coastal environments and calls for a deeper understanding of its applications for corrosion prevention.

Objective To evaluate the feasibility of AIMS65 score combined with neutrophil-to-lymphocyte ratio(NLR) in predicting in-hospital death from esophageal and gastric variceal bleeding(EGVB) in cirrhosis in order to assist clinical patient management. Methods A total of 326 patients with EGVB due to liver cirrhosis who were hospitalized in Taizhou People′s Hospital Affiliated to Nanjing Medical University from January 2018 to October 2022 were selected as the study subjects.According to the outcome during hospitalization, 33 patients were divided into death group and 293 patients into survival group.The clinical data of all patients were collected by electronic medical records, and multivariate logistic regression was used to screen the independent risk factors for in-hospital death in cirrhotic EGVB patients;ROC curve was used to analyze the clinical value of AIMS65 score combined with NLR at admission in predicting in-hospital death in EGVB patients. Results Univariate analysis showed that there were significant differences in albumin, NLR, and AIMS65 scores between the two groups(P

Objective To explore the effect of myotubularin-related protein 6 (MTMR6) on the invasion of hepatocellular carcinoma cell line HepG2 and the potential molecular mechanism. Methods By analyzing the sequencing results of liver cancer tissues and adjacent tissues in Gene Expression Omnibus (GEO) database, MTMR6 gene was screened out, and Spearman analysis was used to analyze the correlation of MTMR6 and pathway in the Cancer Genome Atlas (TCGA) database. Finally, the interaction between MTMR6 and signaling pathway proteins was analyzed with Genemania database. Then the expression of MTMR6 in human normal liver cell line LO-2 and hepatoma cell lines Huh-7 and HepG2 were measured and compared among the cell lines. Then HepG2 cells was selected as the study object. After MTMR6 gene was knocked down or over-expressed in HepG2 cells, Transwell assay was employed to observe invasion ability, and Western blotting was adopted to detect the expression of MTMR6, PI3K, p-PI3K, AKT, p-AKT, mTOR, p-mTOR MMP-2 and MMP-9. Results The expression of MTMR6 was significantly higher in the hepatocellular carcinoma tissues than the paracancer tissues, and it was in a positive linear correlation with PI3K/AKT/mTOR signaling pathway (P < 0.01), showing interaction with PI3K, AKT and mTOR. The expression level of MTMR6 was significantly higher in the HepG2 cells than the LO-2 and Huh-7 cells (P < 0.01). Over-expression of MTMR6 obviously enhanced invasion ability (P < 0.01), while its knockdown decreased the ability (P < 0.01) in HepG2 cells. Knockdown of MTMR6 gene also resulted in decreased phosphorylation of PI3K, AKT and mTOR, and expression levels of MMP-2 and MMP-9 (P < 0.01), while over-expression of MTMR6 promoted the phosphorylation of PI3K, AKT and mTOR, and up-regulated the expression of MMP-2 and MMP-9 (P < 0.01). In addition, LY294002 (a specific PI3K inhibitor) treatment could block the PI3K/AKT/mTOR pathway and down-regulate the expression of MMP-2 and MMP-9 (P < 0.01), but had no effect on MTMR6 expression. Conclusion MTMR6 may promote the invasion of hepatoma cells through activation of PI3K/AKT/mTOR signaling pathway.

Hypersharpening is one of the fusion-based superresolution approaches in remote sensing that improves the spatial and spectral resolution of a hyperspectral image (HSI) with a low spatial resolution. This requirement is achieved by fusing the HSI with a panchromatic image that has high spatial resolution to generate a newly combined variant, which has high spatial quality and high spectral resolution. While several studies in the literature applied neural networks for hypersharpening, there exist unsolved issues such as how to deeply discover the spatial–spectral correlation and inject geometric details without distortion. To address these issues, we propose a hypersharpening technique by applying a multicomponent-based hierarchical fusion network called (HS-McHF), which hierarchically learns the low and high-frequency spatial–spectral features. We then suggest an optimization model to discover the correlation between low-resolution HSI and high-resolution panchromatic images and solve it by stochastic gradient descent through a neural network. Moreover, we decrease the band overlapping in the initial HSI by combining a deconvolution model to prevent spectral distortion and reduce the noise in the panchromatic geometric details injection by deploying an encoder–decoder network. Our extensive experiments demonstrate that the HS-McHF provides superior efficiency compared to state-of-the-art fusion based superresolution approaches.

Abstract The nasal cavity harbors diverse microbiota that contributes to human health and respiratory diseases. However, whether and to what extent the host genome shapes the nasal microbiome remains largely unknown. Here, by dissecting the human genome and nasal metagenome data from 1401 healthy individuals, we demonstrated that the top three host genetic principal components strongly correlated with the nasal microbiota diversity and composition. The genetic association analyses identified 63 genome-wide significant loci affecting the nasal microbial taxa and functions, of which 2 loci reached study-wide significance (p

Abstract The Illumina HiSeq platform has been a commonly used option for bacterial genome sequencing. Now the BGI DNA nanoball (DNB) nanoarrays platform may provide an alternative platform for sequencing of bacterial genomes. To explore the impact of sequencing platforms on bacterial genome assembly, quality assessment, sequence alignment, functional annotation, mutation detection, and metagenome mapping, we compared genome assemblies based on sequencing of cultured bacterial species using the HiSeq 2000 and BGISEQ-500 platforms. In addition, simulated reads were used to evaluate the impact of insert size on genome assembly. Genome assemblies based on BGISEQ-500 sequencing exhibited higher completeness and fewer N bases in high GC genomes, whereas HiSeq 2000 assemblies exhibited higher N50. The majority of assembly assessment parameters, sequences of 16S rRNA genes and genomes, numbers of single nucleotide variants (SNV), and mapping to metagenome data did not differ significantly between platforms. More insertions were detected in HiSeq 2000 genome assemblies, whereas more deletions were detected in BGISEQ-500 genome assemblies. Insert size had no significant impact on genome assembly. Taken together, our results suggest that DNBSEQ platforms would be a valid substitute for HiSeq 2000 for bacterial genome sequencing.

Abstract Background Intervertebral disc degeneration (IDD) is considered an important pathological basis for spinal degenerative diseases. Tissue engineering is a powerful therapeutic strategy that can effectively restore the normal biological properties of disc units. In this study, hydrogels loaded with growth/differentiation factor 5 (GDF5) and stem cells were combined to provide an effective strategy for nucleus pulposus regeneration. Methods Nucleus pulposus stem cells (NPSCs) were obtained by low-density inoculation and culture, and their stem cell characteristics were verified by flow cytometry and a tri-lineage-induced differentiation experiment. A decellularized nucleus pulposus matrix (DNPM) and chitosan hybrid hydrogel was prepared, and GDF5-loaded poly(lactic-co-glycolic acid) (PLGA) microspheres were incorporated into the hydrogels to obtain a composite hydrogels with GDF5-loaded microspheres. Taking bone marrow mesenchymal stem cells (BMSCs) as a reference, the effect of composite hydrogels with GDF5-loaded microspheres on the chondrogenic differentiation of NPSCs was evaluated. A model of intervertebral disc degeneration induced by acupuncture on the tail of rats was constructed, and the repair effect of composite hydrogels with GDF5-loaded microspheres combined with NPSCs on IDD was observed. Results Stem cell phenotype identification, stemness gene expression and tri-lineage-induced differentiation confirmed that NPSCs had characteristics similar to those of BMSCs. The rat DNPM and chitosan hybrid hydrogels had good mechanical properties, and the GDF5-loaded microspheres sustainably released GDF5. NPSCs grew normally in the composite hydrogels and gradually expressed a chondrocyte phenotype. Animal experiments showed that the composite hydrogels with GDF5-loaded microspheres combined with NPSCs effectively promoted nucleus pulposus regeneration and that the effect of the hydrogels on the repair of IDD was significantly better than that of BMSCs. Conclusion GDF5-loaded microspheres combined with DNPM/chitosan composite hydrogels can effectively promote the differentiation of NPSCs into nucleus pulposus-like cells and effectively preventIDD.

Lignans are a powerful weapon for plants to resist stresses and have diverse bioactive functions to protect human health. Elucidating the mechanisms of stereoselective biosynthesis and response to stresses of lignans is important for the guidance of plant improvement. Here, we identified the complete pathway to stereoselectively synthesize antiviral (−)-lariciresinol glucosides in Isatis indigotica roots, which consists of three-step sequential stereoselective enzymes DIR1/2, PLR, and UGT71B2. DIR1 was further identified as the key gene in respoJanuary 2024nse to stresses and was able to trigger stress defenses by mediating the elevation in lignan content. Mechanistically, the phytohormone-responsive ERF transcription factor LTF1 colocalized with DIR1 in the cell periphery of the vascular regions in mature roots and helped resist biotic and abiotic stresses by directly regulating the expression of DIR1. These systematic results suggest that DIR1 as the first common step of the lignan pathway cooperates with PLR and UGT71B2 to stereoselectively synthesize (−)-lariciresinol derived antiviral lignans in I. indigotica roots and is also a part of the LTF1-mediated regulatory network to resist stresses. In conclusion, the LTF1-DIR1 module is an ideal engineering target to improve plant Defenses while increasing the content of valuable lignans in plants.

Biomass energy, recognized for its potential to supplant traditional energy sources, finds a promising alternative in Miscanthus. Particularly, M. lutarioriparius, a species within the Miscanthus genus, stands out as the most suitable for application. This study focuses on the impact of harvest timing on the biomass and combustion quality of M. lutarioriparius in the coastal wetlands of Dongting Lake, Hunan. A comprehensive field experiment was conducted in this area. The findings reveal several key trends: Firstly, the biomass yield of M. lutarioriparius decreased from 18.42 tons per hectare to 11.25 tons per hectare with the delayed harvest. there is a notable increase in the stem-to-leaf ratio of M. lutarioriparius, primarily attributed to leaf shedding. Secondly, the water content had dropped to 22.33% by the time of harvest in February of the following year. Ash content decreased from 4.55% to 2.17% from October to February of the following year. The calorific value, cellulose and lignin content increased with delayed harvest, while hemicellulose content decreased. Notably, the calorific value exhibits a significant positive correlation with cellulose and lignin content, and a markedly negative correlation with hemicellulose. Thirdly, most mineral elements in the plant demonstrate a significant decrease with delayed harvesting, except for potassium (K) and calcium (Ca), which initially increase and then decrease. The ash content shows a positive correlation with the contents of potassium (K) and silicon (Si). Despite a reduction in biomass yield due to delayed harvesting, combustion quality significantly improves. Therefore, it is concluded that optimal fuel quality is achieved when M. lutarioriparius is harvested in February of the following year.

IntroductionFaecalibacterium is one of the most abundant bacteria in the gut microbiota of healthy adults, highly regarded as a next-generation probiotic. However, the functions of Faecalibacterium genomes from cultured strains and the distribution of different species in populations may differ among different sources.MethodsWe here performed an extensive analysis of pan-genomes, functions, and safety evaluation of 136 Faecalibacterium genomes collected from 10 countries.ResultsThe genomes are clustered into 11 clusters, with only five of them were characterized and validly nomenclated. Over 80% of the accessory genes and unique genes of Faecalibacterium are found with unknown function, which reflects the importance of expanding the collection of Faecalibacterium strains. All the genomes have the potential to produce acetic acid and butyric acid. Nine clusters of Faecalibacterium are found significantly enriched in the healthy individuals compared with patients with type II diabetes..DiscussionThis study provides a comprehensive view of genomic characteristic and functions and of culturable Faecalibacterium bacterium from human gut, and enables clinical advances in the future.

Vaginitis, characterized by pathogenic invasion and a deficiency in beneficial lactobacilli, has recognized lactobacilli supplementation as a novel therapeutic strategy. However, due to individual differences in vaginal microbiota, identifying universally effective Lactobacillus strains is challenging. Traditional methodologies for probiotic selection, which heavily depend on extensive in vitro experiments, are both time-intensive and laborious. The aim of this study was to pinpoint possible vaginal probiotic candidates based on whole-genome screening. We sequenced the genomes of 98 previously isolated Lactobacillus strains, annotating their genes involved in probiotic metabolite biosynthesis, adherence, acid/bile tolerance, and antibiotic resistance. A scoring system was used to assess the strains based on their genomic profiles. The highest-scoring strains underwent further in vitro evaluation. Consequently, two strains, Lactobacillus crispatus LG55-27 and Lactobacillus gasseri TM13-16, displayed an outstanding ability to produce d-lactate and adhere to human vaginal epithelial cells. They also showed higher antimicrobial activity against Gardnerella vaginalis, Escherichia coli, Candida albicans, Staphylococcus aureus, and Pseudomonas aeruginosa compared to reference Lactobacillus strains. Their resilience to acid and bile environments highlights the potential for oral supplementation. Oral and vaginal administration of these two strains were tested in a bacterial vaginosis (BV) rat model at various doses. Results indicated that combined vaginal administration of these strains at 1 × 106 CFU/day significantly mitigated BV in rats. This research offers a probiotic dosage guideline for vaginitis therapy, underscoring an efficient screening process for probiotics using genome sequencing, in vitro testing, and in vivo BV model experimentation.

Background: Rheumatoid arthritis (RA) is a chronic inflammatory disease that leads to progressive joint damage. Circular RNA (circRNA) can regulate the inflammatory response of fibroblast-like synoviocytes (FLSs) in RA, influencing the disease progression. Paeoniflorin (PF) is the main active ingredient extracted from Paeonia lactiflora and is known for its anti-inflammatory effect. This study aims to explore the potential mechanisms by which hsa_circ_009012 and PF regulate the inflammatory response in RA. Methods: RNA expression of hsa_circ_009012, has-microRNA-1286 (miR-1286), toll-like receptor 4 (TLR4), NOD-like receptor thermal protein domain associated protein 3 (NLRP3) was assessed by real-time quantitative polymerase chain reaction (RT-qPCR) or western blotting (WB). Cell inflammation markers (TNF-α, IL-1β, IL-6) were assessed by RT-qPCR and immunofluorescence (IF). Counting Kit-8 (CCK-8) assay, flow cytometry, and transwell assay were utilized to test cell viability, cell cycle distribution, and migration. Results: Hsa_circ_009012 was highly expressed in RA-FLS. Hsa_circ_009012 over-expression facilitated the inflammation in RA-FLS and was closely associated with the miR-1286/TLR4 axis. Paeoniflorin inhibited inflammation and the expression of hsa_circ_009012 and TLR4, while upregulating the expression of miR-1286 in RA-FLS. Moreover, the upregulation of hsa_circ_009012 reversed the repressive effect of paeoniflorin on RA-FLS progression. Conclusion: Paeoniflorin inhibits the inflammation of RA-FLS via mediating the hsa_circ_009012/miR-1286/TLR4/NLRP3 axis.

BackgroundStomolophus meleagris envenomation causes severe cutaneous symptoms known as jellyfish dermatitis. The potential molecule mechanisms and treatment efficiency of dermatitis remain elusive because of the complicated venom components. The biological activity and molecular regulation mechanism of Troxerutin (TRX) was firstly examined as a potential treatment for jellyfish dermatitis. MethodsWe examined the inhibit effects of the TRX on tentacle extract (TE) obtained from S. meleagris in vivo and in vitro using the mice paw swelling models and corresponding assays for Enzyme-Linked Immunosorbent Assay (ELISA) Analysis, cell counting kit-8 assay, flow cytometry, respectively. The mechanism of TRX on HaCaT cells probed the altered activity of relevant signaling pathways by RNA sequencing and verified by RT-qPCR, Western blot to further confirm protective effects of TRX against the inflammation and oxidative damage caused by TE. ResultsTE significantly induced the mice paw skin toxicity and accumulation of inflammatory cytokines and reactive oxygen species in vivo and vitro. Moreover, a robust increase in the phosphorylation of mitogen-activated protein kinase (MAPKs) and nuclear factor-kappa B (NF-κB) signaling pathways was observed. While, the acute cutaneous inflammation and oxidative stress induced by TE were significantly ameliorated by TRX treatment. Notablly, TRX suppressed the phosphorylation of MAPK and NF-κB by initiating the nuclear factor erythroid 2-related factor 2 signaling pathway, which result in decreasing inflammatory cytokine release. ConclusionTRX inhibits the major signaling pathway responsible for inducing inflammatory and oxidative damage of jellyfish dermatitis, offering a novel therapy in clinical applications.

Abstract The Lachnospiraceae family holds promise as a source of next‐generation probiotics, yet a comprehensive delineation of its diversity is lacking, hampering the identification of suitable strains for future applications. To address this knowledge gap, we conducted an in‐depth genomic and functional analysis of 1868 high‐quality genomes, combining data from public databases with our new isolates. This data set represented 387 colonization‐selective species‐level clusters, of which eight genera represented multilineage clusters. Pan‐genome analysis, single‐nucleotide polymorphism (SNP) identification, and probiotic functional predictions revealed that species taxonomy, habitats, and geography together shape the functional diversity of Lachnospiraceae. Moreover, analyses of associations with atherosclerotic cardiovascular disease (ACVD) and inflammatory bowel disease (IBD) indicated that several strains of potentially novel Lachnospiraceae species possess the capacity to reduce the abundance of opportunistic pathogens, thereby imparting potential health benefits. Our findings shed light on the untapped potential of novel species enabling knowledge‐based selection of strains for the development of next‐generation probiotics holding promise for improving human health and disease management.

In this study, oral supplementation of stachyose significantly improved HFD-induced MetS symptom, including overweight, hyperlipidemia, hepatic steatosis and system-wide inflammation. The subsequent analysis revealed that stachyose supplement significantly enhanced the integrity of the intestinal epithelial barrier and effectively reversed the gut microbiota dysbiosis, as evidenced by improvements in gut microbial gene richness, microbiota composition, and functional characteristics. The abundance of butyrate-producing strains, such as Bacteroides faecis, Butyomonas faecalis, Parabacteroides distasonis and Phocaeicola coprophilus is significantly increased, concurrently with the activation of the PPAR-γ signaling pathway. The findings of our study suggest that stachyose exhibits potential as a prebiotic agent for the prevention of gut microbiota dysbiosis and the intestinal epithelial barrier disruption in obese. The results of our study suggest that stachyose demonstrates potential as a prebiotic agent for mitigating gut microbiota dysbiosis and preserving the integrity of the intestinal epithelial barrier in individuals with MetS.

This study aims to utilize silicon carbide (SiC) powder and phenol-formaldehyde resin (PF) as the primary raw materials to prepare phenol-formaldehyde coated silicon carbide composite (PCSC) using a thermal coating method. Then, single-factor experiment and orthogonal experiment were used to optimize the process parameters of selective laser sintering (SLS). Finally, SiC precursors were formed using optimized process parameters with excellent density and dimensional accuracies. The results show that the best combination of process parameters is: laser power of 25 W, scanning speed of 1.7 m/s, scanning spacing of 0.12 mm and layer thickness of 0.16 mm.

This paper presents a new analytical solution to investigate the mechanism of transient confined-unconfined flow in a confined aquifer induced by pumping with a large rate during mine drainage. The study focuses on understanding the impact of non-Darcian effect on flow towards a fully penetrated pumping well. The nonlinear relationship between specific discharge and the hydraulic gradient is described using Izbash's equation. A novel approximate method is developed to linearize the mathematical model, and the solution is derived using the Boltzmann transform. The proposed solution is validated by comparing it with previous works. The findings indicate that increased non-Darcian index, quasi-hydraulic conductivity, and specific storage have negatively affect the development of the unconfined region and aquifer drawdown, as greater turbulence flow accelerates recharge to the pumping well. Drawdown is found to be sensitive to the non-Darcian index, quasi-hydraulic conductivity, while it is unaffected by specific yield and specific storage. The conclusions provide valuable insights for mine drainage and the application of geological and hydrological conditions.

Maritime communications with sea surface reflections and sea wave occlusions are susceptible to jamming attacks due to the wide geographical area and intensive wireless communication services. Unmanned Aerial Vehicles (UAVs) help relay messages to improve communication performance, but the relay policy that depends on the rapidly changing maritime environments is difficult to optimize. In this paper, a reinforcement learning-based UAV relay policy for maritime communications is proposed to resist jamming attacks. Based on previous transmission performance, the relay location, the received power of the transmitted signal and the received jamming power, this scheme optimizes the UAV trajectory and relay power to save the energy consumption and decrease the Bit-Error-Rate (BER) of the maritime signals. A deep reinforcement learning-based scheme is also proposed, which designs a deep neural network with dueling architecture to further improve the communication performance and computational complexity. The performance bounds regarding the signal to interference plus noise ratio, energy consumption and the communication utility are provided based on the Nash equilibrium of the game against jamming, and the computational complexity of the proposed schemes is analyzed. Simulation results show that the proposed schemes improve the energy efficiency and decrease the BER compared with the benchmark.

Abstract Background While there have been previous studies on the surgical efficacy of patients with redundant nerve roots (RNRs), a persistent issue is that some patients continue to experience redundancy even after surgery. Furthermore, the clinical significance of RNRs remains unclear. Notably, there is a lack of research regarding RNRs within the context of oblique lumbar interbody fusion (OLIF) combined with percutaneous internal fixation. Therefore, the primary objective of this study is to investigate the correlation between RNRs and clinical outcomes following OLIF combined with percutaneous internal fixation. Methods Eighty-seven patients diagnosed with lumbar spinal stenosis (LSS) who underwent single-segment OLIF combined with percutaneous internal fixation were categorized into three groups. Group 1 comprised patients with positive RNRs both before and after the operation. Group 2 included patients with positive RNRs preoperatively but negative RNRs postoperatively. Group 3 consisted of patients with consistently negative RNRs before and after the operation. Comprehensive patient data were collected, including operation time, intraoperative blood loss, and any recorded complications. Radiographic parameters, both pre- and post-operative, were assessed, encompassing the number of stenosis segments, disc height (DH), lumbar lordotic angle, dural sac cross-sectional area, and the placement of the fusion cage. Furthermore, the Visual Analogue Scale was applied to gauge back and leg pain, while the Oswestry Disability Index was employed to appraise daily living activities. A comparative analysis was carried out among the three patient groups. Results In this study, all 87 LSS patients successfully underwent surgery. Among them, 35 patients (40.2%) showed preoperative MRI assessment indicating positive RNRs. In the postoperative MRI assessment, 14 of these patients maintained positive RNRs status, and they were grouped into Group 1. The remaining 21 patients saw a transition to negative RNRs status and were included in Group 2. Among the 52 patients who had preoperative MRI assessments showing negative RNRs, their postoperative RNRs status remained negative, forming Group 3. All patients received follow-up, which ranged from 8 to 18 months, and no complications occurred during this period. In this study, the postoperative efficacy and parameters such as DH and Dural Sac CSA significantly improved compared to preoperative values for all 87 patients. Patients with preoperative RNRs had more stenosis segments, smaller dural sac CSA, and more severe symptoms. In all three groups, postoperative efficacy scores significantly improved compared to preoperative scores. Group 2 patients had their fusion cages placed more in the middle, while Group 1 patients had their fusion cages more anteriorly located. Group 2 patients exhibited greater recovery in dural sac CSA postoperatively compared to Group 1 patients. Additionally, Group 2 patients had better ODI efficacy scores compared to Group 1 patients. Conclusions Irrespective of the presence or absence of RNRs, patients experienced improvement after undergoing OLIF combined with percutaneous internal fixation. Preoperative RNRs appear to be linked to multi-segmental lumbar spinal stenosis, a reduction in dural sac CSA, and symptom severity. Patients with negative postoperative RNRs demonstrated better treatment efficacy. Furthermore, the placement of the fusion cage appears to have a significant impact on postoperative efficacy and RNRs outcomes.

Pinus is an economically and ecologically important genus whose members are dominant components globally in low-latitude mountainous and mid-latitude temperate forests. Pinus species richness is currently concentrated in subtropical mid-low latitudes of the Northern Hemisphere, differing from the latitudinal diversity gradient mostly recognized in woody angiosperms. How the present pattern was developing in Earth's past is still poorly studied, particularly in eastern Asia. Here, a new fossil species, Pinus shengxianica sp. nov. is described based on a fossil seed cone from the Late Miocene Shengxian Formation in Zhejiang, southeast China. A co-occurring cone is recognized as a known fossil species, Pinus speciosa Li. Extensive comparison of extant and fossil members of Pinus suggests P. shengxianica shares a striking cone similarity to Pinus merkusii and Pinus latteri (subsection Pinus) from tropical Southeast Asia in having annular bulges around the umbo on the apophysis. The morphological resemblance indicates these two extant low-latitude pines probably possess a close affinity with the present newly-discovered P. shengxianica and originated from East Asian mid-low latitude ancestors during this generic re-diversification in the Miocene. This scenario is consistent with the evolutionary trajectory reflected by the pine fossil history and molecular data, marking the Miocene as a key period for the origin and evolution of most extant pines globally. The co-occurrences of diverse conifers and broadleaved angiosperms preferring diverse niches demonstrate Late Miocene eastern Zhejiang was one of the hot spots for coniferophyte diversity and hosted a needled-broadleaved mixed forest with complex vegetation structure and an altitudinal zonation.

The giant jellyfish Nemopilema nomurai sting can cause local and systemic reactions; however, comparative analysis of the tentacle extract (TE) and nematocyst venom extract (NV), and its toxicity, mechanism, and potential intervention are still limited. This study compared venom from TE and NV for their composition, toxicity, and efficacy in vitro and in vivo used RAW264.7 cells and ICR mice. A total of 239 and 225 toxin proteins were identified in TE and NV by proteomics, respectively. Pathological analysis revealed that TE and NV caused heart and liver damage through apoptosis, necrosis, and inflammation, while TE exhibited higher toxicity ex vivo and in vivo. Biochemical markers indicated TE and NV elevated creatine kinase, lactatedehydrogenase, and aspartate aminotransferase, with the TE group showing a more significant increase. Transcriptomics and Western blotting indicated both venoms increased cytokines expression and MAPK signaling pathways. Additionally, 1 mg/kg PACOCF3 (the phospholipase A2 inhibitor) improved survival from 16.7% to 75% in mice. Our results indicate that different extraction methods impact venom activities, tentacle autolysis preserves toxin proteins and their toxicity, and PACOCF3 is a potential antidote, which establishes a good extraction method of jellyfish venom, expands our understanding of jellyfish toxicity, mechanism, and provides a promising intervention.

Hemolytic activity assessment is a widely used method to evaluate the toxicity of marine organisms, including jellyfish. However, there are some challenges associated with testing hemolytic activity. In this study, four methods were employed to compare the hemolytic activity of jellyfish tentacle extract (TE). Firstly, a microplate reader was used to compare the mouse hemoglobin solution at three different wavelengths (415 nm, 541 nm, and 576 nm), and the most sensitive wavelength was selected for further experiments. Secondly, photomicrograph counting was used to determine the number of complete red blood cells in the field of view. Thirdly, a microplate reader was used to test hemolytic activity in a 96-well plate at 415 nm. Fourthly, a Bicinchoninic Acid (BCA) kit was used to test the concentration of hemoglobin in the solution. Finally, a UV-Vis Spectrophotometer was used to test hemolytic activity at 415 nm. Among the three wavelengths tested, the absorption value was most sensitive at 415 nm. The photomicrograph counting method was able to reflect changes in the shape of Red Blood Cells (RBCs). The microplate reader method may exhibit deviations when the solution concentration is high, while external factors could influence the BCA kit when the toxin concentration in the experimental group is low. The spectrophotometer method was found to be relatively accurate and sensitive to changes. When optimizing the method, it is important to consider the applicability of the Beer-Lambert law and the concentration of solutions.

The genus Bifidobacterium widely exists in human gut and has been increasingly used as the adjuvant probiotics for the prevention and treatment of diseases. However, the functional differences of Bifidobacterium genomes from different regions of the world remain unclear. We here describe an extensive study on the genomic characteristics and function annotations of 1512 genomes (clustered to 849 non-redundant genomes) of Bifidobacterium cultured from human gut. The distribution of some carbohydrate-active enzymes varied among different Bifidobacterium species and continents. More than 36% of the genomes of B. pseudocatenulatum harbored biosynthetic gene clusters of lanthipeptide-class-iv. 99.76% of the cultivated genomes of Bifidobacterium harbored genes of bile salt hydrolase. Most genomes of B. adolescentis, and all genomes of B. dentium harbored genes involved in gamma-aminobutyric acid synthesis. B. longum subsp. infantis were characterized harboring most genes related to human milk oligosaccharide utilization. Significant differences between the distribution of antibiotic resistance genes among different species and continents revealed the importance to use antibiotics precisely in the clinical treatment. Phages infecting Bifidobacterium and horizontal gene transfers occurring in genomes of Bifidobacterium were dependent on species and region sources, and might help Bifidobacterium adapt to the environment. In addition, the distribution of Bifidobacterium in human gut was found varied from different regions of the world. This study represents a comprehensive view of characteristics and functions of genomes of cultivated Bifidobacterium from human gut, and enables clinical advances in the future.

Marginal lands, such as those with saline soils, have potential as alternative resources for cultivating dedicated biomass crops used in the production of renewable energy and chemicals. Optimum utilization of marginal lands can not only alleviate the competition for arable land use with primary food crops, but also contribute to bioenergy products and soil improvement. Miscanthus sacchariflorus and M. lutarioriparius are prominent perennial plants suitable for sustainable bioenergy production in saline soils. However, their responses to salt stress remain largely unexplored. In this study, we utilized 318 genotypes of M. sacchariflorus and M. lutarioriparius to assess their salt tolerance levels under 150 mM NaCl using 14 traits, and subsequently established a mini-core elite collection for salt tolerance. Our results revealed substantial variation in salt tolerance among the evaluated genotypes. Salt-tolerant genotypes exhibited significantly lower Na+ content, and K+ content was positively correlated with Na+ content. Interestingly, a few genotypes with higher Na+ levels in shoots showed improved shoot growth characteristics. This observation suggests that M. sacchariflorus and M. lutarioriparius adapt to salt stress by regulating ion homeostasis, primarily through enhanced K+ uptake, shoot Na+ exclusion, and Na+ sequestration in shoot vacuoles. To evaluate salt tolerance comprehensively, we developed an assessment value (D value) based on the membership function values of the 14 traits. We identified three highly salt-tolerant, 50 salt-tolerant, 127 moderately salt-tolerant, 117 salt-sensitive, and 21 highly salt-sensitive genotypes at the seedling stage by employing the D value. A mathematical evaluation model for salt tolerance was established for M. sacchariflorus and M. lutarioriparius at the seedling stage. Notably, the mini-core collection containing 64 genotypes developed using the Core Hunter algorithm effectively represented the overall variability of the entire collection. This mini-core collection serves as a valuable gene pool for future in-depth investigations of salt tolerance mechanisms in Miscanthus.

The explosion of digital technology and the Internet has elevated big data as a critical driver of progress in various fields, including sports education in higher education institutions. This article explores the application of structured data mining to refine sports education, beginning with a decision tree algorithm for student sports data analysis. It then employs the Apriori algorithm to explore gender-based sports information correlations with teaching levels and the K-means algorithm to measure the enhancement in sports teaching quality pre and post-technology adoption. Findings reveal a strong association between improved teaching quality and student physical well-being, highlighted by the College of Physical Education’s top teaching quality score of 10.0. Initially, most teaching quality evaluations were in the “poor” to “good” range (81.2%), shifting significantly to “excellent” and “good” (78.4%) after the intervention. This study evidences the importance of data mining in revolutionizing physical education, significantly boosting educational quality.

Abstract Since sequential recommendation can effectively capture the user’s dynamic interaction pattern according to the user’s historical interaction, it plays one of the important roles in the field of recommendation systems. Nowadays, the sequential recommendation models based on deep learning approaches have achieved good results in reality due to their excellent performance in information retrieval and filtering. However, there are still some challenges that still should be overcome of them. A major disadvantage of them is that they are only considering users’ short-time interests while ignoring their long-term preferences. Moreover, they are incapable of considering the influence of time information in the original interaction sequence, which could be helpful to fully extract various patterns via the different temporal embedding forms. Therefore, this paper proposes a novel model named multi-temporal sequential recommendation model based on the fused learning preference named MTSR-FLP in short. In the proposed framework, this paper adopts an MLP-based state generation module to consider the user’s long-term preferences and the short-time interests simultaneously. In particular, the proposed MTSR-FLP model designs a global representation learning approach to obtain the user’s global preference, and a local representation learning approach to capture the users’ local preference via its historical information. Moreover, the proposed model develops a multiple temporal embedding scheme to encode the positions of user–item interactions of a sequence, in which multiple kernels are utilized for the absolute or relative timestamps to establish unique embedding matrices. Finally, compared with other advanced sequence recommendation models on five public real-world datasets, the experimental results show that the proposed MTSR-FLP model has improved the performance of HR@10 from the 6.68% through 31.10% and NDCG@10 from the 8.60% through 42.54%.

Abstract The oral cavity harbors highly diverse communities of microorganisms. However, the number of isolated species and high-quality genomes is limited. Here we present a Cultivated Oral Bacteria Genome Reference (COGR), comprising 1089 high-quality genomes based on large-scale aerobic and anaerobic cultivation of human oral bacteria isolated from dental plaques, tongue, and saliva. COGR covers five phyla and contains 195 species-level clusters of which 95 include 315 genomes representing species with no taxonomic annotation. The oral microbiota differs markedly between individuals, with 111 clusters being person-specific. Genes encoding CAZymes are abundant in the genomes of COGR. Members of the Streptococcus genus make up the largest proportion of COGR and many of these harbor entire pathways for quorum sensing important for biofilm formation. Several clusters containing unknown bacteria are enriched in individuals with rheumatoid arthritis, emphasizing the importance of culture-based isolation for characterizing and exploiting oral bacteria.

In today’s society, there is an urgent need to help users better access information that they are interested in, as there is an increasing amount of news and messages available with the development of the Internet. Many existing methods involve directly inputting text into a pre-trained model, which limits the effectiveness of text feature extraction. The personalized news recommendation model discussed in this article is a model that can enhance feature extraction from news articles. It consists of a candidate news module, a historically accessed news module, and an access prediction module. Using news titles that accurately summarize news content, a model with double multi-head attention mechanisms and double residual structures (DDM) is utilized to better capture the features of news articles historically accessed by users, thereby achieving an improved recommendation functionality. The candidate news module aims to help the model learn representations of news that users are likely to select from the news titles. The user historical click news module primarily serves to enable the model to learn personalized representations of users from news they have previously browsed. The model has been tested on MIND-small. The AUC reached 0.6665, the MRR reached 0.3205, the nDCG@5 reached 0.3532, and the nDCG@10 reached 0.4158. The results indicate this model has achieved good results in the downstream tasks of preprocessing news-title texts.

The 21st century has marked the dawn of an aging population. China’s aging process ranks first worldwide. The country has recognized the gravity of this demographic shift and implemented strategies to address it at the national level. A vast majority of elderly Chinese individuals (approximately 90%) aspire to age in their own homes. Smart homes, endowed with cutting-edge digital technologies, such as AI, the Internet of Things, and big data, hold vast potential for enabling this vision. However, acceptance of smart home products and services among elderly individuals in China remains low. The main reason is that the related products and services fail to effectively alleviate the perceived risk of this population in the R&D process of related products and services, and there is a lack of effective measurement methods. To holistically assess the potential obstacles faced by elderly individuals using smart home products and services, this study targeted individuals aged 45–60 years in China. This study aimed to develop a comprehensive perceived risk scale specific to smart homes for this demographic. Initially, this study identified key risk dimensions and corresponding measurement items through a rigorous literature review, user interviews, and expert consultations. Subsequently, it ensured the reliability and validity of each dimension and its corresponding observation variables through preliminary research, exploratory factor analysis, and confirmatory factor analysis. This approach allowed for a comprehensive understanding of the challenges faced by future elderly individuals when utilizing smart home products and services, thus enabling the development of more effective solutions. The scale encompassed ten factors and seventy measurement items, including Privacy and Security Risk (seven items), Physical Risk (seven items), Technological Risk (nine items), Performance Risk (seven items), Service Risk (nine items), Financial Risk (five items), Psychological Risk (seven items), Industry and Market Risk (six items), Social Support Risk (six items), and Policy and Legal risk (seven items). The measurement scale developed in this study represents a groundbreaking first attempt to create a systematic scale for assessing the perceived risks associated with smart homes for the elderly in China. It not only enables professionals, businesses, and manufacturers to avoid or reduce barriers in the R&D process of related products and services, facilitating smart home industry growth and enhancing user adoption, but also serves as a universal reference for the potential obstacles that digital technology may encounter in addressing aging-related issues, which has significant theoretical value and practical importance.

The porous architectures of oxygen cathodes are highly desired for high-capacity lithium–oxygen batteries (LOBs) to support cathodic catalysts and provide accommodation for discharge products. However, controllable porosity is still a challenge for laminated cathodes with cathode materials and binders, since polymer binders usually shield the active sites of catalysts and block the pores of cathodes. In addition, polymer binders such as poly(vinylidene fluoride) (PVDF) are not stable under the nucleophilic attack of intermediate product superoxide radicals in the oxygen electrochemical environment. The parasitic reactions and blocking effect of binders deteriorate and then quickly shut down the operation of LOBs. Herein, the present work proposes a binder-free three-dimensional (3D) porous graphene (PG) cathode for LOBs, which is prepared by the self-assembly and the chemical reduction of GO with triblock copolymer soft templates (Pluronic F127). The interconnected mesoporous architecture of resultant 3D PG cathodes achieved an ultrahigh capacity of 10,300 mAh g−1 for LOBs. Further, the cathodic catalysts ruthenium (Ru) and manganese dioxide (MnO2) were, respectively, loaded onto the inner surface of PG cathodes to lower the polarization and enhance the cycling performance of LOBs. This work provides an effective way to fabricate free-standing 3D porous oxygen cathodes for high-performance LOBs.

Objective: This study aimed to investigate the antioxidation and synergistic effect of flavonoid extract of Zanthoxylum armatum DC. Methods: DPPH radical and ABTS radical scavenging methods were used to evaluate the antioxidant activity of flavonoids extract from Z. armatum, and the synergies of flavonoids extract with vitamin C and rutin complex were studied by adding method. Results: The semi-inhibitory concentrations of DPPH free radical and ABTS free radical IC50 were 0.03 and 0.062 mg/mL, respectively, showing strong antioxidant activity. The antioxidant activity of the complex composed of flavonoid extract and vitamin C or rutin was high, and the antioxidant activity of the complex composed of vitamin C and flavonoid extract was higher than that of rutin and flavonoid extract when the ratio was 1∶1. Conclusion: The flavonoids of Z. armatum have strong antioxidant activity, and their complex composed of vitamin C and rutin can improve the scavenging capacity of DPPH free radical and ABTS free radicals. The extracted flavonoids have antioxidant synergistic effect, and can be used as a potential source of natural antioxidants.

Abstract In recent years, nuclear magnetic resonance (NMR) logging has become increasingly prevalent in the characterization of rock properties such as porosity, permeability, saturation, and pore size distribution. However, interpreting such properties accurately from NMR logging data is challenging for some reservoirs. In particular, the impact of salinity, viscosity, and saturation on NMR measurements is not always clear, which can lead to inaccuracies in the resulting data interpretation. Properly accounting for these factors is essential in order to obtain accurate and reliable measurements for effective characterization of subsurface formations. This study utilized a random walk technique to simulate the NMR response of homogeneous sandstones using 3D CT images and conducted a sensitivity analysis under various salinity, crude oil viscosity, and water saturation conditions. The results indicate that the T2 relaxation time slightly shifts toward the short relaxation direction as the salinity of the formation water increases. In addition, longer echo intervals result in a more significant forward shift in the T2 value than shorter intervals. Whereas, for crude oil, the T2 relaxation time becomes shorter as its viscosity increases. Furthermore, the effect of echo interval on the forward T2 shift is less pronounced for crude oil than it is for formation water. Under water wet conditions, the T2 spectrum of crude oil exhibits a peak at the volume relaxation position. As the water saturation decreases, the left two peaks in the T2 spectrum shift toward shorter relaxation times. Under oil wet conditions, the T2 spectrum exhibits a complex three-peak structure. The method provides a physical basis for interpreting NMR macroscopic responses, and the simulated NMR responses can help identify fluids in reservoirs.

Abstract Hepatocellular carcinoma (HCC) is the most common type of primary hepatic carcinoma, which is a growing public health problem worldwide. One of the main genetic alterations in HCC is the deregulated Wnt/β-catenin signaling, activation of β-catenin is associated with the progression of HCC. In the present study, we aimed to identify novel modulators in controlling β-catenin ubiquitination and stability. USP8 was overexpressed in HCC tissues and correlated with β-catenin protein level. High expression of USP8 indicated poor prognosis of HCC patients. USP8 depletion significantly decreased β-catenin protein level, β-catenin target genes expression and TOP-luciferase activity in HCC cells. Further mechanistic study revealed that the USP domain of USP8 interacted with the ARM domain of β-catenin. USP8 stabilized β-catenin protein via inhibiting K48-specific poly-ubiquitination process on β-catenin protein. In addition, USP8 depletion inhibited the proliferation, invasion and stemness of HCC cells and conferred ferroptosis resistance, which effects could be further rescued by β-catenin overexpression. In addition, the USP8 inhibitor DUB-IN-3 inhibited the aggressive phenotype and promoted ferroptosis of HCC cells through degradation of β-catenin. Thus, our study demonstrated that USP8 activated the Wnt/beta-catenin signaling through a post-translational mechanism of β-catenin. High expression of USP8 promoted the progression and inhibited ferroptosis of HCC. Targeting the USP8 may serve as a promising strategy for patients with HCC.

Abstract Background Pueraria montana var. lobata (kudzu) is an important food and medicinal crop in Asia. However, the phylogenetic relationships between Pueraria montana var. lobata and the other two varieties (P. montana var. thomsonii and P. montana var. montana) remain debated. Although there is increasing evidence showing that P. montana var. lobata adapts to various environments and is an invasive species in America, few studies have systematically investigated the role of the phylogenetic relationships and evolutionary patterns of plastomes between P. montana var. lobata and its closely related taxa. Results 26 newly sequenced chloroplast genomes of Pueraria accessions resulted in assembled plastomes with sizes ranging from 153,360 bp to 153,551 bp. Each chloroplast genome contained 130 genes, including eight rRNA genes, 37 tRNA genes, and 85 protein-coding genes. For 24 newly sequenced accessions of these three varieties of P. montana, we detected three genes and ten noncoding regions with higher nucleotide diversity (π). After incorporated publically available chloroplast genomes of Pueraria and other legumes, 47 chloroplast genomes were used to construct phylogenetic trees, including seven P. montana var. lobata, 14 P. montana var. thomsonii and six P. montana var. montana. Phylogenetic analysis revealed that P. montana var. lobata and P. montana var. thomsonii formed a clade, while all sampled P. montana var. montana formed another cluster based on cp genomes, LSC, SSC and protein-coding genes. Twenty-six amino acid residues were identified under positive selection with the site model. We also detected six genes (accD, ndhB, ndhC, rpl2, rpoC2, and rps2) that account for among-site variation in selective constraint under the clade model between accessions of the Pueraria montana var. lobata clade and the Pueraria montana var. montana clade. Conclusion Our data provide novel comparative plastid genomic insights into conservative gene content and structure of cp genomes pertaining to P. montana var. lobata and the other two varieties, and reveal an important phylogenetic clue and plastid divergence among related taxa of P. montana come from loci that own moderate variation and underwent modest selection.

Abstract Virus‐induced gene silencing (VIGS) is a powerful tool for transient gene functional analysis in plants, especially for monocot species (e.g., grasses) that are recalcitrant to transformation. Despite various VIGS systems that have been developed in different plant species, none was previously available for the bioenergy crop Miscanthus. Here, we report the establishment of an efficient and robust VIGS system mediated by Tobacco Rattle Virus (TRV) in Miscanthus. We first investigated the impact of various factors that may affect gene silencing efficiency using the Miscanthus sinensis Phytoene Desaturase (MsPDS) gene as a visual indicator of photobleaching. Then, we optimized the TRV‐elicited VIGS procedure using an orthogonal experimental design with four factors (sprout size, Agrobacterium concentration, vacuum infiltration time, and co‐incubation time) each at three levels. The following led to the highest silencing efficiency (~76%): inoculation of germinating seedlings (1.0–2.0 mm), Agrobacterium tumefaciens culture grown to optical density at 600 nm (OD600) of 0.4, vacuum infiltration for 90 min, and co‐incubation for 5 h. The VIGS system established was applicable for both M. sinensis and M. lutarioriparius, with comparable gene silencing efficiency. We verified the efficacy of the VIGS system via the functional characterization of the role of a MYB transcription factor, MsMYB112, in salt stress tolerance. Expression of MsMYB112 was successfully knocked down using the VIGS system, and this led to compromised salt tolerance in the silenced Miscanthus plants. The TRV‐based VIGS system established may, therefore, substantially facilitate functional genomic studies in Miscanthus.

In this research, V-type porous starch (VPS) with high oil absorption capacity was obtained by enzymatic hydrolysis of V-type granular starch (VGS) in ethanol aqueous solution. Firstly, the reaction condition of V-type granular starch (VGS) was optimized with oil absorption capacity and V-type crystal crystallinity as indexes. In 50% ethanol, aqueous solution the oil absorption capacity of VGS obtained by reaction 100 ℃ was the highest (176.23%). Then, the influence of enzymatic hydrolysis time, temperature and speed on the degree of hydrolysis, yield and oil absorption capacity of VPS were investigated. The optimal preparation conditions of VPS were obtained through single factor and orthogonal experiments: enzymatic hydrolysis time 4 h, temperature 25 ℃, and enzymatic hydrolysis speed 300 r/min. The oil absorption capacity of the optimal VPS was the highest (550.28%). Finally, the difference in the crystalline structure, granular morphology, specific surface area and bulk density were compared among normal maize starch (NMS), traditional A-type porous starch, VGS and VPS. VPS exhibited a unique sponge shaped irregular dense honeycomb structure, the highest specific surface area (39.94 m2/g) and the lowest bulk density (0.19 g/cm3), which were the reasons for its higher oil absorption capacity.

Abstract Background Nano-Zinc oxide (Nano-ZnO) has been increasingly applied in agriculture, industry and biomedicine. However, the genotoxic effects of Nano-ZnO and the underlying mechanisms remain incompletely clear. Methods Human bronchial epithelial cell line (HBE) was used to observe the effects of Nano-ZnO on DNA damage repair-related proteins and epithelial mesenchymal transition (EMT) by Western blotting. Then, CRISPR/cas9-based technique was used to create p53 knockout (p53-KO) cell line. RNA-seq analysis was performed to uncover the circular RNA (circRNA) profile after Nano-ZnO treatment in p53-KO cells compared with p53 wild-type (p53-wt) cells. LC–MS/MS was used to discover the potential binding proteins of circRNA_0085439 in the p53 deficiency background after Nano-ZnO treatment. Nano-ZnO-induced DNA damage and EMT were also investigated in vivo by instillation of Nano-ZnO (50 µg/mouse). Results Nano-ZnO exposure caused DNA damage and EMT at both in vitro and in vivo background, which was reflected by increased DNA damage associated proteins such as ATM and ATR and γ H2AX. p53 expression increased at the early stage post Nano-ZnO treatment decreased later. RNA-seq assay showed a highest increase of circRNA_0085439 expression in p53-KO cells compared with the p53-wt cells after Nano-ZnO exposure. Silencing of p53 expression promoted its translocation of circRNA_0085439 from cytoplasm to nucleus leading to the formation of circRNA_0085439/Ku70 complex resulting in the decreased expression of Ku70 protein. In addition, increased EMT markers, N-cadherin and Vimentin, was observed in lung epithelial cells and in mouse lungs at day 7 after Nano-ZnO exposure. Conclusions This study unraveled the epigenetic mechanisms underlying Nano-ZnO-induced DNA damage and EMT. The effect of Nano-ZnO-induced DNA damage through p53/circRNA_0085439/Ku70 pathway likely contribute to Nano-ZnO-induced cell cytotoxicity and apoptosis. Our findings will provide information to further elucidate the molecular mechanisms of Nano-ZnO-induced cytotoxicity and genotoxicity.

Abstract Intermittent fasting (IF) is a promising paradigm for weight loss which has been shown to modulate the gut microbiota based on 16S rRNA gene amplicon sequencing. Here, 72 Chinese volunteers with a wide range of body mass index (BMI) participated in a three-week IF program during which an average loss of 3.67 kg body weight accompanied with improved clinical parameters was observed irrespective of initial anthropometric and gut microbiota status. Fecal samples were collected before and after the intervention and subjected to shotgun metagenomic sequencing. De novo assembly yielded 2934 metagenome-assembled genomes (MAGs). Profiling revealed significant enrichment of Parabacteroides distasonis and Bacteroides thetaiotaomicron after the intervention, with inverse correlations between their relative abundances and parameters related to obesity and atherosclerotic cardiovascular diseases (ASCVD). MAGs enriched after the intervention showed high richness and diversity of carbohydrate-active enzymes, with an increased relative abundances of genes related to succinate production and glutamate fermentation.

Background: Reducing Ca2+ content in the sarcoplasmic reticulum (SR) through ryanodine receptors (RyRs) by calcin is a potential intervention strategy for the SR Ca2+ overload triggered by β-adrenergic stress in acute heart diseases. Methods: OpiCal-PEG-PLGA nanomicelles were prepared by thin film dispersion, of which the antagonistic effects were observed using an acute heart failure model induced by epinephrine and caffeine in mice. In addition, cardiac targeting, self-stability as well as biotoxicity were determined. Results: The synthesized OpiCa1-PEG-PLGA nanomicelles were elliptical with a particle size of 72.26 nm, a PDI value of 0.3, and a molecular weight of 10.39 kDa. The nanomicelles showed a significant antagonistic effect with 100 % survival rate to the death induced by epinephrine and caffeine, which was supported by echocardiography with significantly recovered heart rate, ejection fraction and left ventricular fractional shortening rate. The FITC labeled nanomicelles had a strong membrance penetrating capacity within 2 h and cardiac targeting within 12 h that was further confirmed by immunohistochemistry with a self-prepared OpiCa1 polyclonal antibody. Meanwhile, the nanomicelles can keep better stability and dispersibility in vitro at 4 °C rather than 20 °C or 37 °C, while maintain a low but stable plasma OpiCa1 concentration in vivo within 72 h. Finally, no obvious biotoxicities were observed by CCK-8, flow cytometry, H&E staining and blood biochemical examinations. Conclusion: Our study also provide a novel nanodelivery pathway for targeting RyRs and antagonizing the SR Ca2+ disordered heart diseases by actively releasing SR Ca2+ through RyRs with calcin.

Abstract Oxaliplatin (OXA) resistance is a major clinic challenge in hepatocellular carcinoma (HCC). Ferroptosis is a kind of iron‐dependent cell death. Triggering ferroptosis is considered to restore sensitivity to chemotherapy. In the present study, we found that USP20 was overexpressed in OXA‐resistant HCC cells. High expression of USP20 in HCC was associated with poor prognosis. USP20 contributes OXA resistance and suppress ferroptosis in HCC. Pharmacological inhibition or knockdown of USP20 triggered ferroptosis and increased the sensitivity of HCC cells to OXA both in vitro and in vivo. Coimmunoprecipitation results revealed that the UCH domain of USP20 interacted with the N terminal of SLC7A11. USP20 stabilized SLC7A11 via removing K48‐linked polyubiquitination of SLC7A11 protein at K30 and K37. Most importantly, DNA damage‐induced ATR activation was required for Ser132 and Ser368 phosphorylation of USP20. USP20 phosphorylation at Ser132 and Ser368 enhanced its stability and thus conferred OXA and ferroptosis resistance of HCC cells. Our study reveals a previously undiscovered association between OXA and ferroptosis and provides new insight into mechanisms regarding how DNA damage therapies always lead to therapeutic resistance. Therefore, targeting USP20 may mitigate the development of drug resistance and promote ferroptosis of HCC in patients receiving chemotherapy.

The pore solution composition (OH−, Na+, K+, Ca2+ and SO42-, S2O32-, S2- concentrations) of hardened concrete materials, including binary systems of PC mixed with a single SCM and with two SCMs, was investigated. Based on database comprising more than 400 entries with more than 80 parameters, machine learning (ML) is applied to predict ion concentrations. Catboost model is the optimal model. The concentrations of OH−, Na+, K+ were predicted with high accuracy (R2 of 0.92–0.95). The prediction accuracy of S is low, could also reaches a R2 of 0.79. But the prediction accuracy of linear regression model is very low, with R2 of 0.18–0.7. PC_MgO, PC_Na2O, PC_K2O and SCM_SiO2 rank high in the characteristic importance analysis for predicting the concentration of OH−, Na+, K+. Compared with the classical pore solution prediction methods (Taylor's and NIST algorithm), the ML model is more accurate. Due to the use of more data and kinds of methods, the ML prediction results in this paper are also better than Cristhiana's ML model. These ML models can be used to predict pore solution of more than 28 d old normal PC concrete with silica fume, fly ash, slag, limestone or quartz powder, but PC or SCM with high phosphorus oxide is not suitable.