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To investigate the effects of epigallocatechin-3-gallate-bovine bone protein (EGCG-BBP) on the protein structure and storage oxidative stability of emulsified meat products, the effects of adding different concentrations of EGCG-BBP on the physicochemical and structural properties of myofibrillar protein (MP) in raw meat, and on the oxidative stability of meatballs were investigated. The results showed that the addition of 0.8% EGCG-BBP resulted in the highest content of sulfhydryl group of MP (4.06 nmol/mg protein) in the minced meat, while the carbonyl group and surface hydrophobicity values were lowest, effectively improving the antioxidant capacity of the emulsified meat products. Fourier-transform infrared spectroscopy analysis showed that the wave number corresponding to the peak of the amide A band of MP was significantly increased in the minced meat supplemented with EGCG-BBP compared with the control group, indicating corresponding changes in the secondary structure of MP. The fluorescence spectra showed that there was a significant red shift in the strongest MP wavelength in the control group with a prolongation of the storage time. However, this shift was significantly reduced as the concentration of EGCC-BBP increased. This suggested that the addition of EGCG-BBP changed the tertiary structure of MP. Furthermore, analysis of oxidation indices during storage of the emulsified meatballs showed that the addition of 0.8% EGCG-BBP significantly reduced the values of both PV values and TBARS in the meatballs, thereby improving their oxidative stability. In summary, EGCG-BBP can significantly alter both the secondary and tertiary structures of MP and has good oxidation resistance. This indicates its potential to enhance the quality of emulsified meat products and provides a new choice for the application of antioxidant emulsifiers in meat products.

Abstract The precise separation of multicomponent signals encounters numerous challenges due to the complexity of signals and widespread interference. Synchrosqueezing Transform (SST) is one of the important technologies for improving the accurate separation of multicomponent signals, but it faces challenges in terms of the difficulty and effectiveness of squeezing. This paper introduces a multicomponent signal separation method based on innovative Fractional Synchrosqueezing Transform (FrSST). FrSST rearranges along the fractional frequency axis, improving the accuracy of time–frequency ridges and, consequently, enhancing the precision of multicomponent signal separation. In the signal reconstruction process, chirp multiplication and energy rearrangement compensate for chirp bases’ effects, boosting energy concentration and reconstruction potential. Utilizing improved ridges from FrSST ensures effective signal reconstruction. Simulation comparisons demonstrate that, with varying SNRs from − 5 to 15 dB, the reconstructed components based on FrSST exhibit favorable approximation to the original signal components. Furthermore, as the sample size increases, the proposed algorithm shows satisfactory computational efficiency.

Abstract Cold stress can impact plant biology at both the molecular and morphological levels. We cultivated two different types of tobacco seedlings using distinct seeding methods, observing significant differences in their cold tolerance at 4 °C. After 12 h cold stress, shallow water seeding cultivation treatment demonstrates a relatively good growth state with slight wilting of the leaves. Tobacco grown using the float system exhibited short, thick roots, while those cultivated through shallow water seeding had elongated roots with more tips and forks. After cold stress, the shallow water seeding cultivation treatment demonstrated higher antioxidant enzyme activity, and lower malondialdehyde (MDA) content. Transcriptome analysis was performed on the leaves of these tobacco seedlings at three stages of cold treatment (before cold stress, after cold stress, and after 3 days of recovery). Upon analyzing the raw data, we found that the shallow water seeding cultivation treatment was associated with significant functional enrichment of nicotinamide adenine dinucleotide (NAD) biosynthesis and NAD metabolism before cold stress, enrichment of functions related to the maintenance of cellular structure after cold stress, and substantial functional enrichment related to photosynthesis during the recovery period. Weighted gene co-expression network analysis (WGCNA) was conducted, identifying several hub genes that may contribute to the differences in cold tolerance between the two tobacco seedlings. Hub genes related to energy conversion were predominantly identified in shallow water seeding cultivation treatment during our analysis, surpassing findings in other areas. These include the AS gene, which controls the synthesis of NAD precursors, the PED1 gene, closely associated with fatty acid β-oxidation, and the RROP1 gene, related to ATP production. Overall, our study provides a valuable theoretical basis for exploring improved methods of cultivating tobacco seedlings. Through transcriptome sequencing technology, we have elucidated the differences in gene expression in different tobacco seedlings at three time points, identifying key genes affecting cold tolerance in tobacco and providing possibilities for future gene editing.

Background/purpose: Developmental defects of enamel (DDE) is a dental disease with a high prevalence and no effective means of prevention. One of the major causes of DDE is infection, but the pathogenesis is still unclear. Melatonin is known for its anti-inflammatory and mineralization-promoting activities. However, the effects of melatonin on inflammation-induced DDE remain unknown. Here, we investigated the pathogenesis and potential therapeutic targets of inflammation-induced DDE. Materials and methods: First, the effect of lipopolysaccharide-induced inflammation in pregnant mice on the enamel mineralization of the offspring was detected by 3D X-ray microscope analysis, immunohistochemical assays, and quantitative real-time polymerase chain reaction (qRT-PCR). Then, the ameloblastic differentiation ability of ameloblast lineage cells (ALCs) in macrophage conditioned medium (CM) was detected. Subsequently, ameloblastic mineralization after melatonin administration was studied both in vivo and in vitro. The underlying mechanism of melatonin was investigated by RNA sequencing and small interfering RNA transfection. Results: Enamel mineralization was decreased in the inflammatory environment both in vivo and in vitro. Furthermore, melatonin treatment ameliorated these defects. RNA sequencing analysis revealed that regulator of G protein signaling 2 (Rgs2) was downregulated in the inflammation group, whereas it was upregulated after the addition of melatonin. Further studies showed that Rgs2 knockdown resulted in decreased ameloblastic mineralization in ALCs. After Rgs2 knockdown of ALCs in M1-CM with melatonin, the effect of melatonin-mediated attenuation of DDE was greatly reduced. Conclusion: Our results demonstrate that melatonin ameliorates inflammation-induced DDE by upregulating RGS2, suggesting that RGS2 is a potential therapeutic target for inflammation-induced DDE.

Underwater concrete structure crack detection and structural health condition assessment based on image processing is a challenging task. The complex underwater environment and severe image degradation seriously affect the accuracy of crack detection. To solve these problems, a monocular vision and image-enhanced fractal-based fractal science based on computer vision and image processing techniques are proposed to carry out a non-contact detection study of underwater concrete cracks. In this study, a four-level structural health condition was established to assist in underwater crack measurement and safety assessment. The box-counting method was used as a practical tool to calculate the fractal dimension. To verify the effective distance of the algorithm, three distances of 0.5 m,0.8 m, and 1.2 m were set. The results show that the method proposed in this study can effectively detect cracks in submerged concrete members within 0.6 m and help managers correctly determine the health of the structure using the fractal dimension.

Abstract Background Leaf morphology plays a crucial role in photosynthetic efficiency and yield potential in crops. Cigar tobacco plants, which are derived from common tobacco (Nicotiana tabacum L.), possess special leaf characteristics including thin and delicate leaves with few visible veins, making it a good system for studying the genetic basis of leaf morphological characters. Results In this study, GWAS and QTL mapping were simultaneously performed using a natural population containing 185 accessions collected worldwide and an F2 population consisting of 240 individuals, respectively. A total of 26 QTLs related to leaf morphological traits were mapped in the F2 population at three different developmental stages, and some QTL intervals were repeatedly detected for different traits and at different developmental stages. Among the 206 significant SNPs identified in the natural population using GWAS, several associated with the leaf thickness phenotype were co-mapped via QTL mapping. By analyzing linkage disequilibrium and transcriptome data from different tissues combined with gene functional annotations, 7 candidate genes from the co-mapped region were identified as the potential causative genes associated with leaf thickness. Conclusions These results presented a valuable cigar tobacco resource showing the genetic diversity regarding its leaf morphological traits at different developmental stages. It also provides valuable information for novel genes and molecular markers that will be useful for further functional verification and for molecular breeding of leaf morphological traits in crops in the future.

Abstract Background With the surge in genomic data driven by advancements in sequencing technologies, the demand for efficient bioinformatics tools for sequence analysis has become paramount. BLAST-like alignment tool (BLAT), a sequence alignment tool, faces limitations in performance efficiency and integration with modern programming environments, particularly Python. This study introduces PxBLAT, a Python-based framework designed to enhance the capabilities of BLAT, focusing on usability, computational efficiency, and seamless integration within the Python ecosystem. Results PxBLAT demonstrates significant improvements over BLAT in execution speed and data handling, as evidenced by comprehensive benchmarks conducted across various sample groups ranging from 50 to 600 samples. These experiments highlight a notable speedup, reducing execution time compared to BLAT. The framework also introduces user-friendly features such as improved server management, data conversion utilities, and shell completion, enhancing the overall user experience. Additionally, the provision of extensive documentation and comprehensive testing supports community engagement and facilitates the adoption of PxBLAT. Conclusions PxBLAT stands out as a robust alternative to BLAT, offering performance and user interaction enhancements. Its development underscores the potential for modern programming languages to improve bioinformatics tools, aligning with the needs of contemporary genomic research. By providing a more efficient, user-friendly tool, PxBLAT has the potential to impact genomic data analysis workflows, supporting faster and more accurate sequence analysis in a Python environment.

Abstract Layer Hall effect (LHE), initially discovered in the magnetic topological insulator MnBi2Te4 film, expands the Hall effect family and opens a promising avenue for layertronics applications. In this study, we present an innovative ferroelectric bilayer model to attain a tunable quantum anomalous layer Hall effect (QALHE). This model comprises two ferromagnetic orbital-active Dirac monolayers stacked antiferromagnetically, accompanied by out-of-plane electric polarization. The interplay between the layer-locked Berry curvature monopoles and the intrinsic out-of-plane electric polarization leads to layer-polarized near-quantized anomalous Hall conductance. Using first-principles calculations, we have identified a promising material for this model, namely FeS bilayer. Our calculations demonstrate that the intrinsic out-of-plane electric polarization in the Bernal-stacked FeS bilayer can induce QALHE by regulating the layer-locked Berry curvature of FeS monolayers. Importantly, the intrinsic electric field can be reversed through interlayer sliding. The discovery of ferroelectrically modulated QALHE paves the way for the integrability and non-volatility of layertronics, offering exciting prospects for future applications.

Aspergillus niger is a highly versatile fungal strain utilized in industrial production. The expression levels of recombinant genes in A. niger can be enhanced by increasing the copy number. Nevertheless, given the prolonged gene editing cycle of A. niger, a “one-step” strategy facilitating the simultaneous integration of recombinant genes into multiple genomic loci would provide a definitive advantage. In our previous study, a visual multigene editing system (VMS) was designed to knock out five genes, employing a tRNA-sgRNA array that includes the pigment gene albA and the target genes. Building upon this system, hybrid donor DNAs (dDNAs) were introduced to establish a clustered regularly interspaced short palindromic repeats (CRISPR)-based multiplex integration toolkit. Firstly, a CRISPR-Cas9 homology-directed repair (CRISPR-HDR) system was constructed in A. niger by co-transforming the CRISPR-Cas9 plasmid (with a highly efficient sgRNA) and the dDNA, resulting in precise integration of recombinant xylanase gene xynA into the target loci (the β-glucosidase gene bgl, the amylase gene amyA, and the acid amylase gene ammA). Subsequently, the length of homology arms in the dDNA was optimized to achieve 100% editing efficiency at each of the three gene loci. To achieve efficient multiplex integration in A. niger, the CRISPR plasmid pLM2 carrying a sgRNA-tRNA array was employed for concurrent double-strand breaks at multiple loci (bgl, amyA, ammA, and albA). Hybrid dDNAs were then employed for repair, including dDNA1-3 (containing xynA expression cassettes without selection markers) and dDNAalbA (for albA knockout). Among the obtained white colonies (RLM2′), 23.5% exhibited concurrent replacement of the bgl, amyA, and ammA genes with xynA (three copies). Notably, the xynA activity obtained by simultaneous insertion into three loci was 48.6% higher compared to that obtained by insertion into only the bgl locus. Furthermore, this multiple integration toolkit successfully enhanced the expression of endogenous pectinase pelA and Candida antarctica lipase CALB. Hence, the combined application of VMS and the CRISPR-HDR system enabled the simultaneous application of multiple selection markers, facilitating the rapid generation in the A. niger cell factories.

Tillering is an important agronomic trait of rice (Oryza sativa) that affects the number of effective panicles, thereby affecting yields. The phytohormone auxin plays a key role in tillering. Here we identified the high tillering and semi-dwarf 1 (htsd1) mutant with auxin-deficiency root characteristics, such as shortened lateral roots, reduced lateral root density, and enlarged root angles. htsd1 showed reduced sensitivity to auxin, but the external application of indole-3-acetic acid (IAA) inhibited its tillering. We identified the mutated gene in htsd1 as AUXIN1 (OsAUX1, LOC_Os01g63770), which encodes an auxin influx transporter. The promoter sequence of OsAUX1 contains many SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) binding sites, and we demonstrated that SPL7 binds to the OsAUX1 promoter. TEOSINTE BRANCHED1 (OsTB1), a key gene that negatively regulates tillering, was significantly downregulated in htsd1. Tillering was enhanced in the OsTB1 knockout mutant, and the external application of IAA inhibited tiller elongation in this mutant. Overexpressing OsTB1 restored the multi-tiller phenotype of htsd1. These results suggest that SPL7 directly binds to the OsAUX1 promoter and regulates tillering in rice by altering OsTB1 expression to modulate auxin signaling.

ABSTRACTFor kinematic relative positioning users between two moving platforms, limited communication bandwidth and computation ability usually cannot support real-time transmission of high-rate (≥10 Hz) BeiDou Navigation Satellite System (BDS) data. The performance of Ambiguity Resolution (AR) is also a major challenge in signal blocked and loss of lock environments. Based on BDS and Inertial Navigation System (INS) data, we develop a novel mode of INS-aided BDS kinematic relative positioning between two moving platforms, aiming to realize real-time, high-rate and precise positioning with low-cost communication modules in challenge environments. To achieve this goal, the INS-aided high-rate relative kinematic positioning and INS-aided BDS AR re-initialization methods are proposed. In this mode, the baseline bias caused by the different position datum is first defined and resolved. Then, high-rate INS data are assisted to obtain kinematic relative position when real-time kinematic positioning results are unavailable within 1 second. Once the BDS data are available, the predicted relative position is used as additional constraint to facilitate AR re-initialization and improve the kinematic positioning performance. The performance of these methods is discussed in a set of experiments with 1 Hz BDS data and 100 Hz INS data, and compared with the conventional method by sending raw BDS/INS measurements. The results show that the proposed methods can achieve an accuracy of about 5 cm for the INS-aided 100 Hz relative positioning in baseline components and lengths, which is equal to the conventional method, but the transmitted data have been sharply reduced by an average of nearly 80%. With the assistance of INS-predicted baseline constraint, the relative positioning performance has been further improved. The accuracy of baseline length errors is less than 4 cm, and the AR fixing rates keep larger than 95% during the experiment, while the wrongly fixed rates are reduced to less than 0.5%.

Vibrio parahaemolyticus is a gram-negative halophilic bacterium widespread in temperate and tropical coastal waters; it is considered to be the most frequent cause of Vibrio-associated gastroenteritis in many countries. BolA-like proteins, which reportedly affect various growth and metabolic processes including flagellar synthesis in bacteria, are widely conserved from prokaryotes to eukaryotes. However, the effects exerted by BolA-like proteins on V. parahaemolyticus remain unclear, and thus require further investigation. In this study, our purpose was to investigate the role played by BolA-like protein (IbaG) in the pathogenicity of V. parahaemolyticus. We used homologous recombination to obtain the deletion strain ΔibaG and investigated the biological role of BolA family protein IbaG in V. parahaemolyticus. Our results showed that IbaG is a bacterial transcription factor that negatively modulates swimming capacity. Furthermore, overexpressing IbaG enhanced the capabilities of V. parahaemolyticus for swarming and biofilm formation. In addition, inactivation of ibaG in V. parahaemolyticus SH112 impaired its capacity for colonizing the heart, liver, spleen, and kidneys, and reduced visceral tissue damage, thereby leading to diminished virulence, compared with the wild-type strain. Finally, RNA-sequencing revealed 53 upregulated and 71 downregulated genes in the deletion strain ΔibaG. KEGG enrichment analysis showed that the two-component system, quorum sensing, bacterial secretion system, and numerous amino acid metabolism pathways had been altered due to the inactivation of ibaG. The results of this study indicated that IbaG exerts a considerable effect on gene regulation, motility, biofilm formation, and pathogenicity of V. parahaemolyticus. To the best of our knowledge, this is the first systematic study on the role played by IbaG in V. parahaemolyticus infections. Thus, our findings may lead to a better understanding of the metabolic processes involved in bacterial infections and provide a basis for the prevention and control of such infections.

PurposeThe learning subjective well-being of high school students has significant value for their academic achievement and future life development. A growth mindset is one of the key factors affecting the learning subjective well-being of high school students. However, research on the mechanism by which a growth mindset affects learning subjective well-being is still relatively limited. Therefore, the study aims to investigate the impact of a growth mindset on the learning subjective well-being of high school students, as well as the role that achievement motivation and grit play as serial mediators in this relationship.MethodsThis study employed a convenience sampling method to select 708 high school students from Chinese public high schools as participants. The research utilized the Growth Mindset Scale, Achievement Motivation Scale, Grit Scale, and the Learning Subjective Well-being Questionnaire for High School Students to collect data. All data were analyzed using SPSS 26.0, employing Model 6 from Hayes’ SPSS PROCESS macro to test the serial mediation model.ResultsOur results found that (1) high school students’ growth mindset positively predicted their learning subjective well-being. (2) Achievement motivation played a mediating role between a growth mindset and learning subjective well-being among high school students. (3) Grit acted as a mediator between learning subjective well-being and growth mindset among high school students. (4) Achievement motivation and grit served as serial mediators between a growth mindset and learning subjective well-being among high school students.ConclusionA growth mindset can influence the learning subjective well-being of high school students through achievement motivation and grit. Educators can enhance the learning subjective well-being of high school students by implementing intervention strategies that foster a growth mindset, achievement motivation, and grit.

Tobacco, a crop of significant economic importance, was greatly influenced in leaf quality by protein content. However, current processing parameters fail to adequately meet the requirements for protein degradation. Microorganisms possess potential advantages for degrading proteins and enhancing the quality of tobacco leaves, and hold substantial potential in the process of curing. To effectively reduce the protein content in tobacco leaves, thereby improving the quality and safety of the tobacco leaves. In this study, tobacco leaf were used as experimental material. From these, the BSP1 strain capable of effectively degrading proteins was isolated and identified as Bacillus subtilis by 16S rDNA analysis. Furthermore, the mechanisms were analyzed by integrating microbiome, transcriptome, and metabolome. Before curing, BSP1 was applied to the surface of tobacco leaves. The results indicated that BSP1 effectively improves the activity of key enzymes and the content of related substances, thereby enhancing protein degradation. Additionally, protein degradation was achieved by regulating the diversity of the microbial community on the surface of the tobacco leaves and the ubiquitin-proteasome pathway. This study provided new strategies for extracting and utilizing functional strains from tobacco leaves, opening new avenues for enhancing the quality of tobacco leaves.

Background: Male breast cancer (MBC) is a rare malignancy with a worse prognosis than female breast cancer (FBC). Current MBC treatment strategies are based on those for FBC. However, molecular differences between MBC and FBC with respect to prognosis and drug responses remain unclear. Methods: After controlling for confounding factors with propensity score matching (PSM), differences between MBC and FBC were comprehensively analyzed using many types of data: survival, immune microenvironments, sex hormone responses, drug sensitivity, transcriptomes, genomes, epigenomes, and proteomes. Results: Overall survival (OS) and cancer-specific survival (CSS) were both worse for MBC than for FBC. Differentially expressed mRNAs were enriched in numerous cancer-related functions and pathways, with SPAG16 and STOX1 being as the most important prognosis-related mRNAs for MBC. Competing endogenous RNA (ceRNA) and transcription factor (TF)-mRNA regulatory networks contain potential prognostic genes. Nine genes had higher mutation frequencies in MBC than in FBC. MBC shows a comparatively poor response to immunotherapy, with five proteins that promote breast cancer progression being highly expressed in MBC. MBC may be more responsive than FBC to estrogen. We detected six United States Food and Drug Administration (FDA)-approved therapeutic target genes as being differentially expressed between MBC and FBC. Conclusion: The poor prognosis of MBC compared to FBC is due to numerous molecular differences and resulting drug responses.

Background: Diabetic foot is a major complication of diabetes. The bone transverse transport method could be applied in clinics for treatment, which could improve the metabolism of the tissues via lasting distraction forces. However, the process’ specific regulating mechanism is still unknown. Methods: Based on the notion that the healing of bones involves the recruitment of calcium ions, in this study, we established the model of tibial cortex transverse transport (TTT) on rats and then used tissue immunologic detection, such as the double fluorescent staining to explore the expression of the calcium channels’ calcium release-activated calcium modulator 1 (Orai1)/stromal interaction molecule 1 (STIM1), which belong to the store-operated calcium entry (SOCE) signaling pathways on the tissues around the bone transport area. By using the laser capture microdissection (LCM) tool, we acquired samples of tissues around the bone and endeavored to identify pivotal protein molecules. Subsequently, we validated the functions of key protein molecules through in vitro and in vivo experiments. Results: After protein profile analysis, we found the differentially expressed key protein osteopontin (OPN). The in vitro experiments verified that, being stimulated by OPN, the migration, proliferation, and angiogenesis of human umbilical vein endothelial cells (HUVEC) were observed to be enhanced. The activation of Orai1/STIM1 might increase the activity of endothelial nitric oxide synthase (eNOS) and its effect on releasing nitric oxide (NO). Subsequently, the migration and proliferation of the HUVECs are improved, which ultimately accelerates wound healing. These signaling pathway was also observed in the OPN-stimulated healing process of the skin wound surface of diabetic mice. Conclusion: This study identifies the molecular biological mechanism of OPN-benefited the migration and proliferation of the HUVECs and provides ideas for searching for new therapeutic targets for drugs that repair diabetes-induced wounds to replace invasive treatment methods. The translational potential of this article: The OPN is highly expressed in the tissues surrounding the TTT bone transfer area, which may possibly stimulate the activation of eNOS to increase NO release through the SOCE pathway mediated by Orai1/STIM1. This mechanism may play a significant role in the angiogenesis of diabetic foot's wounds promoted by TTT, providing new therapeutic strategies for the non-surgical treatment for this disease.

Abstract Rapeseed (Brassica napus) is one of the important oil crops worldwide. Its production is often threatened by drought stress. Here, we identify a transcription factor (BnaA9.NF-YA7) that negatively regulates drought tolerance through genome-wide association study in B. napus. The presence of two SNPs within a CCAAT cis element leads to downregulation of BnaA9.NF-YA7 expression. In addition, the M63I (G-to-C) substitution in the transactivation domain can activate low level expression of BnaA4.DOR, which is an inhibitory factor of ABA-induced stomatal closure. Furthermore, we determine that Bna.ABF3/4 s directly regulate the expression of BnaA9.NF-YA7, and BnaA9.NF-YA7 indirectly suppresses the expression of Bna.ABF3/4 s by regulation of Bna.ASHH4s. Our findings uncover that BnaA9.NF-YA7 serves as a supplementary role for ABA signal balance under drought stress conditions, and provide a potential molecular target to breed drought-tolerant B. napus cultivars.

Abstract Background Observational studies have suggested that sedentary behaviors and sleep status are associated with frailty. However, it remains unclear whether these associations are causal. Methods Using summary statistics from genome-wide association studies, we evaluated the causal effect of modifiable risk factors, including leisure sedentary behaviors and sleep status on the frailty index (FI) using two-sample univariable and multivariable Mendelian randomization (MR) analyses. Genetic correlations were tested between the correlated traits. Results We identified potential causal associations between the time spent watching television (β = 0.26, 95% confidence interval [CI]: 0.21–0.31, P = 3.98e-25), sleep duration (β = -0.18, 95%CI: -0.26, -0.10; P = 6.04e-06), and daytime napping (β = 0.29, 95%CI: 0.18–0.41, P = 2.68e-07) and the FI based on the inverse-variance-weighted method. The estimates were consistent across robust and multivariate MR analyses. Linkage disequilibrium score regression detected a genetic correlation between the time spent watching television (Rg = 0.43, P = 6.46e-48), sleep duration (Rg = -0.20, P = 5.29e-10), and daytime napping (Rg = 0.25, P = 3.34e-21) and the FI. Conclusions Genetic predispositions to time spent watching television and daytime napping were positively associated with the FI, while sleep duration was negatively associated with the FI. Our findings offer key insights into factors influencing biological aging and suggest areas for interventions to promote healthy aging and slow down the aging process.

Abstract Background Tobacco mosaic virus (TMV) is a widely distributed viral disease that threatens many vegetables and horticultural species. Using the resistance gene N which induces a hypersensitivity reaction, is a common strategy for controlling this disease in tobacco (Nicotiana tabacum L.). However, N gene-mediated resistance has its limitations, consequently, identifying resistance genes from resistant germplasms and developing resistant cultivars is an ideal strategy for controlling the damage caused by TMV. Results Here, we identified highly TMV-resistant tobacco germplasm, JT88, with markedly reduced viral accumulation following TMV infection. We mapped and cloned two tobamovirus multiplication protein 2A (TOM2A) homeologs responsible for TMV replication using an F2 population derived from a cross between the TMV-susceptible cultivar K326 and the TMV-resistant cultivar JT88. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9)-mediated loss-of-function mutations of two NtTOM2A homeologs almost completely suppressed TMV replication; however, the single gene mutants showed symptoms similar to those of the wild type. Moreover, NtTOM2A natural mutations were rarely detected in 577 tobacco germplasms, and CRISPR/Cas9-mediated variation of NtTOM2A led to shortened plant height, these results indicating that the natural variations in NtTOM2A were rarely applied in tobacco breeding and the NtTOM2A maybe has an impact on growth and development. Conclusions The two NtTOM2A homeologs are functionally redundant and negatively regulate TMV resistance. These results deepen our understanding of the molecular mechanisms underlying TMV resistance in tobacco and provide important information for the potential application of NtTOM2A in TMV resistance breeding.

Osteopontin (OPN), a bioactive milk protein in breast milk, has diverse applications in food and medicine. OPN is predominantly extracted from fresh bovine milk; however, the concentration of OPN in bovine milk is exceedingly low. To address these issues, in the present study, we used Komagataella phaffii for the recombinant expression of human OPN (hOPN). The results of host cell screening revealed that X33 cells were more suitable than other K. phaffii hosts for hOPN expression, yielding a titer of 340.5 μg/L. Subsequently, it was observed that the extracellular production of hOPN significantly increased to 8.02 mg/L from strain XPSA01 using the PAOX1 promoter and the hybrid signal peptide PROSCW10-α. Meanwhile, the co-expression of transcription factors, translation factors, and molecular chaperones resulted in a 2.94-fold enhancement in extracellular hOPN production compared with that in XPSA01, resulting in a titer of 23.6 mg/L. Finally, the above strategies were combined to obtain the recombinant strain XPSA01-CP, which achieved titers of 35.6 and 128.5 mg/L hOPN in shake-flask fermentation and 3-L bioreactor, respectively. To the best of our knowledge, this is the highest extracellular yield of hOPN ever reported.

BackgroundIncomplete immune recovery in people living with HIV/AIDS (PLWHA) remains an important clinical challenge with the lack of an effective strategy currently available to restore their T-cell immune response. This study aimed to evaluate the effect of Albuvirtide (ABT) on immune recovery in immunological non-responders (INRs) and attempted to explore potential mechanisms of ABT on the functionality of immune cells.MethodsIn this prospective, open-label, controlled clinical study, participants with incomplete immune reconstitution (continuous ART over 5 years and CD4+T lymphocyte absolute count of

Protein phosphorylation is one of the major molecular mechanisms regulating protein activity and function throughout the cell. Pannexin 1 (PANX1) is a large-pore channel permeable to ATP and other cellular metabolites. Its tyrosine phosphorylation and subsequent activation have been found to play critical roles in diverse cellular conditions, including neuronal cell death, acute inflammation, and smooth muscle contraction. Specifically, the non-receptor kinase Src has been reported to phosphorylate Tyr198 and Tyr308 of mouse PANX1 (equivalent to Tyr199 and Tyr309 of human PANX1), resulting in channel opening and ATP release. Although the Src-dependent PANX1 activation mechanism has been widely discussed in the literature, independent validation of the tyrosine phosphorylation of PANX1 has been lacking. Here, we show that commercially available antibodies against the two phosphorylation sites mentioned above—which were used to identify endogenous PANX1 phosphorylation at these two sites—are nonspecific and should not be used to interpret results related to PANX1 phosphorylation. We further provide evidence that neither tyrosine residue is a major phosphorylation site for Src kinase in heterologous expression systems. We call on the field to re-examine the existing paradigm of tyrosine phosphorylation-dependent activation of the PANX1 channel.

Abstract Background Neuroinflammation and mitochondrial dysfunction play crucial roles in retinal ischemia and reperfusion (IR) injury. Recent studies have identified mitochondrial function as a promising target for immunomodulation. Empagliflozin (EMPA), an anti-diabetic drug, has exhibited great potential as both an anti-inflammatory agent and a protector of mitochondrial health. This study aimed to assess the therapeutic efficacy of EMPA in retinal IR injury. Methods To evaluate the protective effects of EMPA, the drug was injected into the vitreous body of mice post-retinal IR. Single-cell RNA sequencing (scRNA-seq) analysis was conducted to uncover the underlying mechanisms, and the results were further validated through in vivo and in vitro experiments. Results EMPA effectively protected retinal ganglion cells (RGCs) from IR injury by attenuating local retinal inflammation. The scRNA-seq analysis revealed that EMPA downregulated the nucleotide-binding domain and leucine-rich repeat containing protein 3 (NLRP3) signaling pathway and restored mitochondrial dynamics by upregulating the expression of mitochondrial fusion-related genes, Mitofusin 1 (Mfn1) and optic atrophy 1 (Opa1). These findings were further corroborated by Western blotting. In vitro experiments provided additional insights, demonstrating that EMPA suppressed lipopolysaccharide (LPS)-induced cell inflammation and NLRP3 inflammasome activation. Moreover, EMPA enhanced mitochondrial fusion, neutralized mitochondrial reactive oxygen species (mtROS), and restored mitochondrial membrane potential (MMP) in BV2 microglia. Notably, genetic ablation of Mfn1 or Opa1 abolished the anti-inflammatory effects of EMPA. Conclusions Our findings highlight the positive contribution of Mfn1 and Opa1 to the anti-inflammatory therapeutic effect of EMPA. By restoring mitochondrial dynamics, EMPA effectively mitigates microglia-mediated neuroinflammation and prevents RGC loss in retinal IR injury.

Abstract Recent advancements in developing metahyperbolic surfaces through substrate patterning have enabled the realization of highly-directional hyperbolic surface plasmons, but the feasibility of reproducing the same properties in natural hyperbolic two-dimensional (2D) materials is still unexplored. In this study, we expand the possibility of natural 2D materials in achieving electromagnetic scenarios akin to those observed in metahyperbolic surfaces. Natural hyperbolic 2D materials provide inherent advantages for simplicity, predictability, and lower losses compared to meta-surfaces. By employing first-principles calculations, we find that realistic 2D material, specifically the RuOCl2 monolayer, are suitable alternatives to metahyperbolic surfaces. Indeed, RuOCl2 monolayer sustains carrier-density-independent and broadband low-loss hyperbolic responses across the terahertz to ultraviolet spectral range, owning to the highly-anisotropic electronic band structures characterized by quasi-one-dimensional electron gas. These findings shed light on the integration of hyperbolicity in natural 2D materials, opening new avenues for the design and development of optoelectronic devices and nanoscale imaging systems.

Grain filling influences grain size and quality in cereal crops. The molecular mechanisms that regulate grain endosperm development remain elusive. In this study, we characterized a filling-defective and grain width mutant, fgw1, whose mutation increased rice seed width mainly via cell division and expansion in grains. Sucrose contents were higher but starch contents lower in the fgw1 mutant during the grain-filling stage, resulting in inferior endosperm of opaque, white appearance with loosely packed starch granules. Map-based cloning revealed that FGW1 encoded a protein containing DUF630/DUF632 domains, localized in the plasma membrane with preferential expression in the panicle. RNA interference in FGW1 resulted in increased grain width and weight, whereas overexpression of FGW1 led to slightly narrower kernels and better grain filling. In a yeast two-hybrid assay, FGW1 interacted directly with the 14–3–3 protein GF14f, bimolecular fluorescence complementation verified that the site of interaction was the membrane, and the mutated FGW1 protein failed to interact with GF14f. The expression of GF14f was down-regulated in fgw1, and the activities of AGPase, StSase, and SuSase in the endosperm of fgw1 increased similarly to those of a reported GF14f-RNAi. Transcriptome analysis indicated that FGW1 also regulates cellular processes and carbohydrate metabolism. Thus, FGW1 regulated grain formation via the GF14f pathway.

Foodborne mycotoxins (FBMTs) are toxins produced by food itself or during processing and transportation that pose an enormous threat to public health security. However, traditional instrumental and chemical methods for detecting toxins have shortcomings, such as high operational difficulty, time consumption, and high cost, that limit their large-scale applications. In recent years, aptamer-based biosensors have become a new tool for food safety risk assessment and monitoring due to their high affinity, good specificity, and fast response. In this review, we focus on the progress of single-mode and dual-mode aptasensors in basic research and device applications over recent years. Furthermore, we also point out some problems in the current detection strategies, with the aim of stimulating future toxin detection systems for a transition toward ease of operation and rapid detection.

The working environment of permanent magnets in high-speed motors is harsh, because they are seriously affected by multi-physical fields such as temperature, stress, weak magnetic control field, etc., and irreversible demagnetization is easy to occur under the extreme working conditions. In this work, a model of high-speed Interior Permanent magnet Motor is established, the easy demagnetization area of the permanent magnet is simulated by electromagnetic and thermal simulations to determine, and its local anti-demagnetization optimization is performed. Dy/Tb co-infiltration is carried out in the easy demagnetization region of the permanent magnets, and the process of selective zone diffusion is analyzed based on the principle of selective zone grain boundary diffusion. Through microscopic observation and macroscopic magnetic performance test, the effect of local anti-demagnetization performance enhancement of the optimized permanent magnets is verified. The results show that the magnetic properties of the optimized permanent magnets are obviously improved after the optimization of the selected zones. The present approach is important for the follow-up less heavy-rare-earth research of permanent magnet and its application in high-speed motors.

Background:The rapid spread of bacteria with plasmid-mediated resistance to antibiotics poses a serious threat to public health. The search for potential compounds that can increase the antibacterial activity of existing antibiotics is a promising strategy for addressing this issue.Methods:Synergistic activity of the FDA-approved agent oxethazine combined with colistin was investigated in vitro using checkerboard assays and time-kill curves. The synergistic mechanisms of their combination of oxethazine and colistin was explored by fluorescent dye, scanning electron microscopy (SEM) and LC-MS/MS. The synergistic efficacy was evaluated in vivo by the Galleria mellonella and mouse sepsis models.Results:In this study, we found that oxethazine could effectively enhance the antibacterial activity of colistin against both mcr-positive and -negative pathogens, and mechanistic assays revealed that oxethazine could improve the ability of colistin to destruct bacterial outer membrane and cytoplasmic membrane permeability. In addition, their combination triggered the accumulation of reactive oxygen species causing additional damage to the membrane structure resulting in cell death. Furthermore, oxethazine significantly enhanced the therapeutic efficacy of colistin in two animal models.Conclusion:These results suggested that oxethazine, as a promising antibiotic adjuvant, can effectively enhance colistin activity, providing a potential strategy for treating multidrug-resistant bacteria.

Tobacco, a vital economic crop, had its quality post-curing significantly influenced by starch content. Nonetheless, the existing process parameters during curing were inadequate to satisfy the starch degradation requirements. Microorganisms exhibit inherent advantages in starch degradation, offering significant potential in the tobacco curing process. Our study concentrated on the microbial populations on the surface of tobacco leaves and in the rhizosphere soil. A strain capable of starch degradation, designated as BS3, was successfully isolated and identified as Bacillus subtilis by phylogenetic tree analysis based on 16SrDNA sequence. The application of BS3 on tobacco significantly enhanced enzyme activity and accelerated starch degradation during the curing process. Furthermore, analyses of the metagenome, transcriptome, and metabolome indicated that the BS3 strain facilitated starch degradation by regulating surface microbiota composition and affecting genes related to starch hydrolyzed protein and key metabolites in tobacco leaves. This study offered new strategies for efficiently improving the quality of tobacco leaves.

Abstract Background Dysbiosis of gut microbiota is causally linked to impaired host glucose metabolism. We aimed to study effects of the new method of fecal microbiota transplantation, washed microbiota transplantation (WMT), on reducing glycemic variability (GV) in unstable diabetes. Methods Fourteen eligible patients received three allogenic WMTs and were followed up at 1 week, 1 month, and 3 months. Primary outcomes were daily insulin dose, glucose excursions during meal tests, and GV indices calculated from continuous monitoring or self‐monitoring glucose values. Secondary outcomes were multiomics data, including 16S rRNA gene sequencing, metagenomics, and metabolomics to explore underlying mechanisms. Results Daily insulin dose and glucose excursions markedly dropped, whereas GV indices significantly improved up to 1 month. WMT increased gut microbial alpha diversity, beta diversity, and network complexity. Taxonomic changes featured lower abundance of genera Bacteroides and Escherichia‐Shigella, and higher abundance of genus Prevotella. Metagenomics functional annotations revealed enrichment of distinct microbial metabolic pathways, including methane biosynthesis, citrate cycle, amino acid degradation, and butyrate production. Derived metabolites correlated significantly with improved GV indices. WMT did not change circulating inflammatory cytokines, enteroendocrine hormones, or C‐peptide. Conclusions WMT showed strong ameliorating effect on GV, raising the possibility of targeting gut microbiota as an effective regimen to reduce GV in diabetes.

Nature, with its numerous surprising rules, serves as a rich source of creativity for the development of artificial intelligence, inspiring researchers to create several nature-inspired intelligent computing paradigms based on natural mechanisms. Over the past decades, these paradigms have revealed effective and flexible solutions to practical and complex problems. This paper summarizes the natural mechanisms of diverse advanced nature-inspired intelligent computing paradigms, which provide valuable lessons for building general-purpose machines capable of adapting to the environment autonomously. According to the natural mechanisms, we classify nature-inspired intelligent computing paradigms into 4 types: evolutionary-based, biological-based, social-cultural-based, and science-based. Moreover, this paper also illustrates the interrelationship between these paradigms and natural mechanisms, as well as their real-world applications, offering a comprehensive algorithmic foundation for mitigating unreasonable metaphors. Finally, based on the detailed analysis of natural mechanisms, the challenges of current nature-inspired paradigms and promising future research directions are presented.

BackgroundPost-stroke depression is a common complication of stroke, with a high incidence rate and low recognition rate. Many patients do not receive effective intervention at the onset, which affects subsequent treatment outcomes. Post-stroke depression not only impacts the patient's mental well-being but also increases the risk of stroke recurrence and poor prognosis. Therefore, it has become a significant public health concern. Acupuncture has gained significant popularity in the treatment of post-stroke depression. However, there are inconsistent clinical research results regarding its efficacy and safety. This systematic review aims to gather and critically assess all available evidence regarding the effectiveness and safety of acupuncture in the treatment of post-stroke depression in patients.MethodsWe will conduct thorough searches for relevant studies in multiple electronic databases (PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, VIP Database, Wan-fang Data and China Biomedical Database). Our search scope will encompass studies published from the inception of each database until September 2023. To evaluate the potential bias in all the included studies, we will adhere to the guidelines offered in the Cochrane Handbook. The total effective rate will be the primary outcome. To conduct a systematic review, we will employ RevMan 5.4 software.ResultsThis study will obtain efficacy and safety of acupuncture for the treatment of post-stroke depression.ConclusionsThe conclusions of this study will provide evidence-based perspectives that can guide clinical decision-making regarding the practicality and recommended timing of using acupuncture to treat post-stroke depression. Furthermore, this study will help advance the clinical application of acupuncture treatment for post-stroke depression and enhance its efficacy while ensuring patient safety.

Abstract Purpose To investigate the clinical efficacy and safety of combination therapy of hepatic arterial infusion chemotherapy (HAIC) and anti‐programmed cell death protein‐1 (PD‐1) therapy in the treatment of advanced hepatocellular carcinoma (HCC). Methods In this retrospective clinical research, from March 2018 to December 2019, 1158 HCC patients categorized as BCLC stage C were reviewed for eligibility. We utilized propensity score matching (PSM) to mitigate initial disparities between the groups. The evaluation of the best tumor response was conducted in accordance with mRECIST 1.1 criteria. The difference in survival outcomes including overall survival (OS), progression‐free survival (PFS), and objective response rate (ORR) between groups were compared. Results Following the eligibility review, 453 patients underwent a combined treatment of HAIC with PD1 inhibitors (HAIC‐PD1 group), while 221 patients received HAIC monotherapy (HAIC group) met the inclusion criteria and were finally enrolled in this study. In the entire cohort, the HAIC‐PD1 group exhibited significantly prolonged overall survival (median overall survival: 40.4 months vs. 9.7 months, p

Cadmium (Cd) is a dangerous heavy metal with high toxicity that is known to impair development. Astilbin (ASB) is a protective flavonoid compound. We aimed to explore whether ASB can antagonize the myocardial developmental toxicity of Cd exposure. Cd (2 µg) and/or ASB (0.002 µg) were injected into embryonized eggs that were 1 day old. Histological examinations revealed Cd-induced ventricular dilation, reduced wall thickness, and disrupted myocardial fiber connections, while co-administration of ASB mitigated these effects. Electron microscopy confirmed ASB's ability to counteract Cd-induced myocardial cell myofibril damage. Real-time quantitative PCR (QRT-PCR) and western blot (WB) molecular investigations revealed that Cd increased endoplasmic reticulum stress in myocardial tissue and primary cardiomyocytes, as shown by raised expression of stress-related genes (GRP78, XBP1, GRP94, ATF4, ATF6, IRE1, and CHOP). Moreover, Cd disrupted calcium homeostasis, affecting important genes linked to Ca2+ channels and causing an excess of Ca2+ in the cytoplasm. In addition, we detected genes related to development and differentiation-related genes in myocardial tissue and primary cardiomyocytes. The results showed that the downregulation of transcription factors in the IrxA cluster, Mefs, and Tbxs families after Cd exposure indicated that cardiac transcription was hindered and cardiac markers (TnnT2, TnnC1, Gata4, Gata6, and Nkx2–5) were abnormally expressed. ASB successfully mitigated these disturbances. During the cell cycle, primary cardiomyocytes undergo growth arrest in flow cytometry. These results suggest that the maturation and differentiation of cardiomyocytes are inhibited after Cd exposure, and ASB has an antagonistic effect on Cd. The present study indicated that Cd could trigger developmental cardiotoxicity in chicken embryos and primary cardiomyocytes by endoplasmic reticulum stress and Ca2+ overload, respectively, while ASB has an antagonistic effect.

Abstract Purpose To develop a deep learning radiomics of multiparametric magnetic resonance imaging (DLRMM)‐based model that incorporates preoperative and postoperative signatures for prediction of local tumor progression (LTP) after thermal ablation (TA) in hepatocellular carcinoma (HCC). Methods From May 2017 to October 2021, 417 eligible patients with HCC were retrospectively enrolled from three hospitals (one primary cohort [PC, n = 189] and two external test cohorts [ETCs][n = 135, 93]). DLRMM features were extracted from T1WI + C, T2WI, and DWI using ResNet18 model. An integrative model incorporating the DLRMM signature with clinicopathologic variables were further built to LTP risk stratification. The performance of these models were compared by areas under receiver operating characteristic curve (AUC) using DeLong test. Results A total of 1668 subsequences and 31,536 multiparametric MRI slice including T1WI, T2WI, and DWI were collected simultaneously. The DLRMM signatures were extracted from tumor and ablation zone, respectively. Ablative margin, multiple tumors, and tumor abutting major vessels were regarded as risk factors for LTP in clinical model. The AUC of DLRMM model were 0.864 in PC, 0.843 in ETC1, and 0.858 in ETC2, which was higher significantly than those in clinical model (p

Abstract Background Cervical cancer is a common gynecological malignancy. Gene microarray found that TCP11 gene was highly expressed in cervical cancer. However, the effect of TCP11 gene on the proliferation, apoptosis and migration of cervical cancer cells and its underlying molecular mechanisms are unclear. Methods GEPIA database, tissue microarray, western blot and qRT-PCR were used to analyze the expression of TCP11 gene in cervical cancer tissues and cells and its relationship with patients’ survival rate. The cell cycle and apoptosis were detected by flow cytometry, and the expressions of cell cycle and apoptosis related molecules and EMT-related molecules were detected by Western blot and qRT-PCR. Results The results showed that TCP11 gene was highly expressed in cervical cancer tissues and cells compared with normal cervical tissues and cells, and its expression was positively correlated with patients’ survival rate. The results of proliferation and migration assays showed that TCP11 overexpression inhibited the proliferation and migration of HeLa and SiHa cells. The results showed that TCP11 overexpression blocked the cell cycle of HeLa and SiHa cells, decreased the expression of CDK1 and Cyclin B1, and increased the apoptosis and the expression of caspase-3, cleaved-caspase-3 and cleaved-PARP. TCP11 overexpression increased the protein and mRNA expression of EMT-related molecules ZO-1 and E-cadherin. Conversely, TCP11 knockdown promoted the proliferation of HeLa and SiHa cells and the migration of HeLa cells. Conclusions TCP11 overexpression significantly inhibited the occurrence and development of cervical cancer cells, it may be a potentially beneficial biomarker for cervical cancer.

The intermediate temperature deformation behavior and microstructural evolution of a spray formed 7075 Al alloy were performed at temperatures 60–120 °C with strain rates 0.00001–0.1 s−1. The results reveal that the softening mechanism of the intermediate temperature deformation is dynamic recovery (DRV), which can be quantitated by Zener-Hollomon (Z) parameter. The constitutive equation can reasonably predict the stress-strain behavior of the alloy at intermediate temperatures. The activation energy is related to the deformation process and the interactions of the substructure, the average value is 140.1 kJ/mol. The higher efficiency of power dissipation in the processing map is related to the DRV at lower deformation temperatures. The optimum processing domain is located at the temperature of 90–120 °C and strain rate of 0.006–0.1 s−1 with a power dissipation efficiency range of 20%–40%. The processing parameters of the intermediate temperature deformation should be strictly controlled according to the strain. The uniform microstructure of 7075 alloys obtained by spray forming is crucial to overcome the disadvantage of poor deformability. The inferior coordinated deformation facilitates crack propagation at low strain rates. The high-density dislocation provides adequate nucleation sites for dynamic precipitation. The level of dynamic restoration facilitates the transformation from low angle grain boundaries (LAGBs) to high angle grain boundaries (HAGBs). The dislocations move to the grain/sub-grain boundary to form dislocation walls at low temperatures. The contribution to the stress from grain boundary (13.2–17.7 MPa), nanoscale precipitates (65.7–191.5 MPa), and dislocation (5.9–23.4 MPa) increases the difficulty of deformation. The deformation process at higher temperatures (105–120 °C) can be facilitated by the dissolution and coarsening of the nanoscale precipitates, and dislocation motion at the grain boundary.

The super-thick, high-pressure, medium-strong water-rich Luohe Formation aquifer is overlying in the Binchang mining area of Shanxi Province, and the fractures in the overlying rock are developed, it makes the water channel easier to communicate with the aquifer and stope of Luohe Formation, resulting in the increase of water inflow and area in the stope. In order to study the morphological characteristics of water inrush induced by the network of water-conducting channels formed by primary fractures communicating with the aquifer of the thick Luohe Formation under the influence of mining, the solid-flow coupling similar material simulation test was carried out based on the similar simulation physical experiment system of water-sand inrush in overburden rock. The results show that when the working face is advanced to 140 m, the lower strata of the bed separation are broken in advance due to the influence of the primary fractures. The left incomplete bed separation space and the triangular space formed by the right cantilever beam support form the “Z” bed separation space. When the working face is advanced to 160 m, two “Z-type” bed separation spaces are developed in the overlying strata, which are interconnected with the primary fractures and mining-induced fractures to form a water channel network. The form of gushing (leaching) water in the stope changed from ‘ drip-drip and flow-flow-multi-state ’, and the overall gushing (leaching) water volume increased first and then decreased. The water pressure of overlying strata and the advancing distance of the working face show a segmented evolution characteristic of decreasing first and then increasing. The minimum interval and the position of the inflection point of the segmentation increase with the increase of the distance between the monitoring point and the open-off cut. The final water pressure values near the central area of the goaf are greater than the two boundary monitoring points. The analysis results show that the existence of primary fractures promotes the development of water-conducting fracture channel network, accelerates the process of water transport, and induces the formation and development of water gushing (leaching) in the stope. The research results clarify the influence of primary fractures on the distribution characteristics of water conduction channel network and the evolution law of water gushing (leaching) form morphology, and explain the conduction mechanism of thick and high confined aquifer water to stope water inrush.

This paper proposes a conceptual design of an innovative deep-sea aquaculture platform that integrates a steel structural framework and high-density polyethylene (HDPE) floats. It aims to overcome the limitations of prevailing aquaculture equipment, including inadequate resistance to strong wind and waves, complex technologies, and prohibitively high costs. The design scheme and key parameters of the main platform, the netting system, and the mooring systems are presented. Based on the stochastic design wave method, the characteristic load response scenarios and design wave parameters are determined and analyzed. Strength analysis is conducted to assess the structural performance, vulnerabilities, and overall safety of the platform under various characteristic load conditions. The results indicate that the Von Mises stress levels across different sections of the platform conform to the allowable stress thresholds under various characteristic load conditions. However, the stress levels of the platform are notably higher when subjected to characteristic loads associated with vertical shear, vertical bending moments, and torsion about the horizontal axis, which requires further efforts in the design process to enhance the structural safety of the platform. The proposed design methodology and the presented research results can provide a wide range of references for the design and analysis of deep-sea fisheries aquaculture equipment.

As nearshore aquaculture spaces become saturated, the development of fisheries aquaculture for deep sea has become an inevitable trend. This paper proposes an innovative deep-sea aquaculture platform that incorporates a vessel-shaped main structure and a single-point mooring system. The potential flow theory and the Morison equation are utilized to calculate the hydrodynamic loads on the main structure and the netting and mooring systems, respectively. The deformation and force of the netting in current are simulated, and the accuracy of the analytical methods used is validated based on experimental results. The influences of the netting system on the hydrodynamic characteristics of the platform are analyzed. Optimization on the single-point mooring system is conducted under static and dynamic conditions, considering the influences of various mooring parameters, including mooring line length, buoyancy of buoys, and mass of sinkers. The patterns of changes in motion response, mooring line tension, and minimum touchdown length under different mooring parameters are calculated and analyzed. The results indicate that changes in mooring line length have minimal impact on the dynamic response of the platform and mooring system. The addition of appropriate buoys or sinkers can reduce the motion response of the platform and the tension in the mooring lines. Moreover, compared to adding buoys, incorporating sinkers more effectively enhances the overall safety and stability of the platform system.

Geopolymers exhibit broad application prospects, including construction and radiation shielding, which require excellent mechanical performances. However, investigations on the nature of geopolymerization reactions and their consequential impact on mechanical performance are still vague. In this study, the effect of the major factors of Si/Al ratio and curing time on the geopolymerization reaction and flexural strength were studied based on the microstructure evolution and chemical bonding formation analyzed using the SEM, FTIR, peak deconvolution, and XRD methods. The microstructure of geopolymers was transferred from initially layered smooth particles of kaolinite to a 3D network porous structure, corresponding to sodalite. A spectrum exclusive to the geopolymer structure occurred at 973 cm−1, corresponding to the sodium aluminum silicate hydrate (N-A-S-H) links, the integral area of which represents the degree of geopolymerization reaction. Furthermore, a controllable reaction degree was achieved by adjusting the Si/Al ratio and curing time, where the maximum reaction degree of 55% was achieved at a Si/Al ratio of 1.94 when cured for 7 d. The correlation between the flexural strength and reaction degree was found to follow a proportional relationship, achieving a flexural strength of 21.11 MPa with a degree of 45%. This study provides insight into the development of mechanical strength through controlling the reaction process.

In order to find more excellent structural materials resistant to radiation damage, high-entropy alloys (HEAs) have been developed due to their characteristics of limited point defect diffusion such as lattice distortion and slow diffusion. Specially, refractory high-entropy alloys (RHEAs) that can adapt to a high-temperature environment are badly needed. In this study, TiZrHfNbMo0.1 RHEAs are selected for irradiation and nanoindentation experiments. We combined the mechanistic model for the depth-dependent hardness of ion-irradiated metals and the introduction of the scale factor f to modify the irradiation-hardening model in order to better describe the nanoindentation indentation process in the irradiated layer. Finally, it can be found that, with the increase in irradiation dose, a more serious lattice distortion caused by a higher defect density limits the expansion of the plastic zone.

Abstract Background Nuclear Yes1‐associated transcriptional regulator (YAP1) promotes tumor progression. However, the function of cytoplasmic YAP1 in breast cancer cells and its impact on the survival of breast cancer patients remain unclear. Our research aimed to explore the biological function of cytoplasmic YAP1 in breast cancer cells and the possibility of cytoplasmic YAP1 as a predictive marker of breast cancer survival. Methods We constructed cell mutant models, including NLS‐YAP15SA (nuclear localized), YAP1S94A (incapable of binding to the TEA domain transcription factor family) and YAP1S127D (cytoplasmic localized), and used Cell Counting Kit‐8 (CCK‐8) assays, 5‐ethynyl‐2’‐deoxyuridine (EdU) incorporation assays, and Western blotting (WB) analysis to detect cell proliferation and apoptosis. The specific mechanism of cytoplasmic YAP1‐mediated endosomal sorting complexes required for transport III (ESCRT‐III) assembly was studied by co‐immunoprecipitation, immunofluorescence staining, and WB analysis. Epigallocatechin gallate (EGCG) was used to simulate YAP1 retention in the cytoplasm in in vitro and in vivo experiments to study the function of cytoplasmic YAP1. YAP1 binding to NEDD4‐like E3 ubiquitin protein ligase (NEDD4L) was identified using mass spectrometry and was verified in vitro. Breast tissue microarrays were used to analyze the relationship between cytoplasmic YAP1 expression and the survival of breast cancer patients. Results YAP1 was mainly expressed in the cytoplasm in breast cancer cells. Cytoplasmic YAP1 promoted autophagic death of breast cancer cells. Cytoplasmic YAP1 bound to the ESCRT‐III complex subunits charged multivesicular body protein 2B (CHMP2B) and vacuolar protein sorting 4 homolog B (VPS4B), promoting assembly of CHMP2B‐VPS4B and activating autophagosome formation. EGCG retained YAP1 in the cytoplasm, promoting the assembly of CHMP2B‐VPS4B to promote autophagic death of breast cancer cells. YAP1 bound to NEDD4L, and NEDD4L mediated ubiquitination and degradation of YAP1. Breast tissue microarrays revealed that high levels of cytoplasmic YAP1 were beneficial to the survival of breast cancer patients. Conclusions Cytoplasmic YAP1 mediated autophagic death of breast cancer cells by promoting assembly of the ESCRT‐III complex; furthermore, we established a new breast cancer survival prediction model based on cytoplasmic YAP1 expression.

Abstract Background Natural killer (NK) cell-based immunotherapies have demonstrated substantial potential for the treatment of hematologic malignancies. However, its application is limited due to the difficulty in the production of a large number of NK cells in vitro and the insufficient therapeutic efficacy against solid tumors in vivo. Engineered antibodies or fusion proteins targeting activating receptors and costimulatory molecules of NK cells have been developed to encounter these problems. They are mostly produced in mammalian cells with high cost and long processing times. Yeast systems, such as Komagataella phaffii, present a convenient manipulation of microbial systems with the key advantages of improved folding machinery and low cost. Results In this study, we designed an antibody fusion protein scFvCD16A-sc4-1BBL, composed of the single chain variant fragment (scFv) of anti-CD16A antibody and the three extracellular domains (ECDs) of human 4-1BBL in a single-chain format (sc) with the GS linker, aiming to boost NK cell proliferation and activation. This protein complex was produced in the K. phaffii X33 system and purified by affinity chromatography and size exclusion chromatography. The scFvCD16A-sc4-1BBL complex showed comparable binding abilities to its two targets human CD16A and 4-1BB as its two parental moieties (scFvCD16A and monomer ECD (mn)4-1BBL). scFvCD16A-sc4-1BBL specifically stimulated the expansion of peripheral blood mononuclear cell (PBMC)-derived NK cells in vitro. Furthermore, in the ovarian cancer xenograft mouse model, adoptive NK cell infusion combined with intraperitoneal (i.p) injection of scFvCD16A-sc4-1BBL further reduced the tumor burden and prolonged the survival time of mice. Conclusion Our studies demonstrate the feasibility of the expression of the antibody fusion protein scFvCD16A-sc4-1BBL in K. phaffii with favourable properties. scFvCD16A-sc4-1BBL stimulates PBMC-derived NK cell expansion in vitro and improves the antitumor activity of adoptively transferred NK cells in a murine model of ovarian cancer and may serve as a synergistic drug for NK immunotherapy in future research and applications.

Abstract Background Retinal ischemia–reperfusion (RIR) injury refers to an obstruction in the retinal blood supply followed by reperfusion. Although the molecular mechanism underlying the ischemic pathological cascade is not fully understood, neuroinflammation plays a crucial part in the mortality of retinal ganglion cells. Methods Single-cell RNA sequencing (scRNA-seq), molecular docking, and transfection assay were used to explore the effectiveness and pathogenesis of N,N-dimethyl-3β-hydroxycholenamide (DMHCA)-treated mice with RIR injury and DMHCA-treated microglia after oxygen and glucose deprivation/reoxygenation (OGD/R). Results DMHCA could suppress inflammatory gene expression and attenuate neuronal lesions, restoring the retinal structure in vivo. Using scRNA-seq on the retina of DMHCA-treated mice, we provided novel insights into RIR immunity and demonstrated nerve injury-induced protein 1 (Ninjurin1/Ninj 1) as a promising treatment target for RIR. Moreover, the expression of Ninj1, which was increased in RIR injury and OGD/R-treated microglia, was downregulated in the DMHCA-treated group. DMHCA suppressed the activation of the nuclear factor kappa B (NF-κB) pathways induced by OGD/R, which was undermined by the NF-κB pathway agonist betulinic acid. Overexpressed Ninj1 reversed the anti-inflammatory and anti-apoptotic function of DMHCA. Molecular docking indicated that for Ninj1, DMHCA had a low binding energy of − 6.6 kcal/mol, suggesting highly stable binding. Conclusion Ninj1 may play a pivotal role in microglia-mediated inflammation, while DMHCA could be a potential treatment strategy against RIR injury.

Deep‐sea exploration remains a challenging task as the extreme hydrostatic pressure environment, darkness, and suspended sediment launch severely hinder the capability of deep‐sea vehicles. As a complement to underwater camera, tactile perception becomes especially important in situations where machine vision is limited. However, tactile sensors utilized in deep sea, which should be able to detect pressure changes of only hundreds of pascals under high hydrostatic pressure, are still lacking. To tackle the challenge imposed by hydrostatic pressure, a simulated deep‐sea environment flexible sensor (SDEFS) is proposed, consisting of a force sensor array and a bending sensor based on hydrogels for tactile sensing in 50 MPa hydrostatic pressure environments. The force sensor is unaffected by the hydrostatic pressure and achieves high sensitivity of 82.62 N−1 under 100 MPa hydrostatic pressure. The SDEFS is utilized to classify objects based on the difference in hardness. It can accurately classify seven objects on the ground, and three objects in an underwater environment with hydrostatic pressure of 50 MPa, with total recognition accuracies of 98.3% and 96%, respectively. With high force measurement sensitivity and accurate recognition ability under water, the SDEFS is expected to provide very valuable haptic sensing and feedback in deep‐sea exploration.

Penis et testis cervi is one of the precious traditional Chinese medicines (TCMs) in China, which, when authentic, is sourced from species of Cervi nippon Temmink (C. Nippon) and Cervi elaphus Linnaeus (C. elaphus L). Authentication of species of Penis et testis cervi remains technically challenging. This study attempted to modify the species-specific polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) method using restriction enzyme Hpy188III for the identification of Penis et testis cervi species. First, the cytochrome b (Cyt b) and cytochrome c oxidase subunit I (COI) genes in mitochondrial DNA from two authentic species and their counterfeits or adulterants were downloaded and analyzed with bioinformatics tools to establish the phylogenetic tree, then a series of primers were designed, and the restriction enzyme sites were confirmed. Genomic DNA was extracted from all samples by the improved SDS method. Species-specific PCR reactions were optimized to differentiate authentic Sika deer and Red deer Penis et testis cervi from their counterfeits or adulterants. PCR-RFLP using single restriction enzyme Hpy188III was performed in Cyt b regions to identify the species of Sika deer and Red deer. Simultaneously, recombination plasmid carrying the cyt b fragment could be cloned, sequenced, and blasted. The assay specificity and sensitivity were evaluated, and 30 batches of commercial samples were validated. The phylogenetic tree showed that the physico-chemical properties of Sika deer and Red deer variants were similar. Sufficient DNA extracts were extracted from all samples used in the improved SDS method. Establishment and optimization of a species-specific PCR reactions and PCR-RFLP system were undertaken to identify deer species in Penis et testis cervi samples. A specific PCR-RFLP pattern with three distinct products of sizes 155, 113, and 204 bp was exhibited in Sika deer species, and two distinct products of 204 and 268 bp in size were evident in Red deer species, which were significantly different from the pattern of the counterfeits or adulterants. The distinctive PCR-RFLP fingerprint pattern using the restriction enzyme Hpy188III is a rapid and reliable assay for identification of the Penis et testis cervi species.

Summary: As the major channel of aqueous humor outflow, dysfunction of trabecular meshwork (TM) can lead to intraocular pressure elevating, which can trigger primary open-angle glaucoma (POAG). In this study, we use single-cell RNA sequencing (scRNA-seq) technique to build an atlas and further explore the spontaneous POAG and healthy macaques cellular heterogeneity associated with the dysfunction of TM contraction. We built the TM atlas, which identified 14 different cell types. In Beam A, Beam B, Beam C, and smooth muscle cell (SMC) cell types, we first found multiple genes associated with TM contraction (e.g., TPM1, ACTC1, TNNT1), determining their differential expression in the POAG and healthy groups. In addition, the microstructural alterations in TM of POAG non-human primates were observed, which was compact and collapsed. Thus, our study indicated that TPM1 may be a key target for regulating TM structure, contraction function, and resistance of aqueous humor outflow.

Securing maize grain yield is crucial to meet food and energy needs for the future growing population, especially under frequent drought events and elevated CO2 (eCO2) due to climate change. To maximize the kernel setting rate under drought stress is a key strategy in battling against the negative impacts. Firstly, we summarize the major limitations to leaf source and kernel sink in maize under drought stress, and identified that loss in grain yield is mainly attributed to reduced kernel set. Reproductive drought tolerance can be realized by collective contribution with a greater assimilate import into ear, more available sugars for ovary and silk use, and higher capacity to remobilize assimilate reserve. As such, utilization of CO2 fertilization by improved photosynthesis and greater reserve remobilization is a key strategy for coping with drought stress under climate change condition. We propose that optimizing planting methods and mining natural genetic variation still need to be done continuously, meanwhile, by virtue of advanced genetic engineering and plant phenomics tools, the breeding program of higher photosynthetic efficiency maize varieties adapted to eCO2 can be accelerated. Consequently, stabilizing maize production under drought stress can be achieved by securing reproductive success by harnessing CO2 fertilization.

Abstract Background Papillary thyroid microcarcinoma (PTMC) incidence has significantly increased, and some cases still exhibit invasive traits. The entire molecular landscape of PTMC, which can offer hints for the etiology of cancer, is currently absent. Methods We compared our findings with those for PTMC in the TCGA by analyzing the largest study at the current stage of whole exome sequencing and RNA-sequencing data from 64 patients with PTMC. Then, we systematically demonstrated the differences between the two PTMC subtypes based on multi-omics analyses. Additionally, we created a molecular prediction model for the PTMC subtypes and validated them among TCGA patients for individualized integrative assessment. Results In addition to the presence of BRAF mutations and RET fusions in the TCGA cohort, we also discovered a new molecular signature named PTMC-inflammatory that implies a potential response to immune intervention, which is enriched with AFP mutations, IGH@-ext fusions, elevated immune-related genes, positive peroxidase antibody, and positive thyroglobulin antibody. Additionally, a molecular prediction model for the PTMC-inflammatory patients was created and validated among TCGA patients, while the prognosis for these patients is poor. Conclusions Our findings comprehensively define the clinical and molecular features of PTMC and may inspire new therapeutic hypotheses.