Search

Abstract Electromagnetic fields (EMFs) have emerged as an effective treatment for osteoporosis. However, the specific mechanism underlying their therapeutic efficacy remains controversial. Herein, we confirm the pro-osteogenic effects of 15 Hz and 0.4-1 mT low-frequency sinusoidal EMFs (SEMFs) on rat bone marrow mesenchymal stem cells (BMSCs). Subsequent miRNA sequencing reveal that miR-34b-5p is downregulated in both the 0.4 mT and 1 mT SEMFs-stimulated groups. To clarify the role of miR-34b-5p in osteogenesis, BMSCs are transfected separately with miR-34b-5p mimic and inhibitor. The results indicate that miR-34b-5p mimic transfection suppress osteogenic differentiation, whereas inhibition of miR-34b-5p promote osteogenic differentiation of BMSCs. In vivo assessments using microcomputed tomography, H&E staining, and Masson staining show that miR-34b-5p inhibitor injections alleviate bone mass loss and trabecular microstructure deterioration in ovariectomy (OVX) rats. Further validation demonstrates that miR-34b-5p exerts its effects by regulating STAC2 expression. Modulating the miR-34b-5p/STAC2 axis attenuate the pro-osteogenic effects of low-frequency SEMFs on BMSCs. These studies indicate that the pro-osteogenic effect of SEMFs is partly due to the regulation of the miR-34b-5p/STAC2 pathway, which provides a potential therapeutic candidate for osteoporosis.

Abstract Recent advances in cancer research have highlighted the pivotal role of tertiary lymphoid structures (TLSs) in modulating immune responses, particularly in breast cancer (BRCA). Here, we performed an integrated analysis of bulk transcriptome data from over 6000 BRCA samples using biological network-based computational strategies and machine learning (ML) methods, and identified LGALS2 as a key marker within TLSs. Single-cell sequencing and spatial transcriptomics uncover the role of LGALS2 in TLS-associated dendritic cells (DCs) stimulation and reveal the complexity of the tumor microenvironment (TME) at both the macro and micro levels. Elevated LGALS2 expression correlates with prolonged survival, which is associated with a robust immune response marked by diverse immune cell infiltration and active anti-tumor pathways leading to a ‘hot’ tumor microenvironment. The colocalization of LGALS2 with TLS-associated DCs and its role in immune activation in BRCA were confirmed by hematoxylin-eosin (HE), immunohistochemistry (IHC), and in vivo validation analyses. The identification of LGALS2 as a key factor in BRCA not only highlights its therapeutic potential in novel TLS-directed immunotherapy but also opens new avenues in patient stratification and treatment selection, ultimately improving clinical management.

Summary: Nucleus pulposus (NP) resides in hypoxic microenvironment and NP cells (NPCs), primarily reply on glycolysis and producing high levels of lactate. Intracellular lactate drives lysine lactylation (Kla) as a newly epigenetic modification. However, the impact of Kla on NPCs remains unknown. Here, single-cell RNA sequencing (scRNA-seq) data suggested an altered balance between glycolysis and aerobic oxidation in intervertebral disc degeneration (IDD). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis displayed 3510 lactylation sites on 1052 non-histone proteins of NPCs isolated from rat cultured in normoxia and hypoxia. Moreover, there are 18 proteins with 129 Kla sites and 117 Kla sites in 27 proteins exclusively detected in normoxia and hypoxia group, respectively. Bioinformatics analysis displayed that these lactylated proteins are tightly related to ribosome, spliceosome and the VEGFA-VEGFA2 signaling pathway. Together, our study reveals that Kla may play an important role in regulating cellular metabolism of NPCs.

Introduction: In yeast, the cytoplasm-to-vacuole targeting (Cvt) pathway acts as a biosynthetic autophagy-related process, in which vacuolar targeting of hydrolase is mediated by the machineries involved in the selective autophagy. However, the mechanistic insights into vacuolar targeting of hydrolases through the selective autophagy pathway still remain enigmatic in filamentous fungi. Objectives: Our study aims to investigate the mechanisms involved in vacuolar targeting of hydrolases in filamentous fungi. Methods: The filamentous entomopathogenic fungus Beauveria bassiana was used as a representative of filamentous fungi. We identified the homologs of yeast aminopeptidase I (Ape1) in B. bassiana by bioinformatic analyses and characterized their physiological roles by gene function analyses. Pathways for vacuolar targeting of hydrolases were investigated via molecular trafficking analyses. Results: B. bassiana has two homologs of yeast aminopeptidase I (Ape1) which are designated as BbApe1A and BbApe1B. The two homologs of yeast Ape1 contribute to starvation tolerance, development, and virulence in B. bassiana. Significantly, BbNbr1 acts as a selective autophagy receptor to mediate the vacuolar targeting of the two Ape1 proteins, in which BbApe1B interacts with BbNbr1 also directly interacting with BbAtg8, and BbApe1A has an additional requirement of the scaffold protein BbAtg11 that interacts with BbNbr1 and BbAtg8. Protein processing occurs at both terminuses of BbApe1A and only at carboxyl terminus of BbApe1B, which is also dependent on the autophagy-related proteins. Together, the functions and translocation processes of the two Ape1 proteins are associated with autophagy in fungal lifecycle. Conclusion: This study reveals the functions and translocation processes for vacuolar hydrolases in the insect-pathogenic fungi and improves our understandings of the Nbr1-mediated vacuolar targeting pathway in the filamentous fungi.

The conventional operation mode of “power determined by heat” employed by thermal power plants， along with the randomness and volatility of new energy power generation， constrains the peak regulation capacity. The incorporation of thermal storage technology into grid peak regulation can optimize the allocation capacity of the energy system. The fundamental principles， appropriate materials， advantages， disadvantages， and future development trends of three different thermal storage technologies， namely， sensible heat storage （SHS）， latent heat storage （LHS） and chemical energy storage （CES）， are introduced. Furthermore， the strategies of different thermal storage technologies for power system peak regulation at home and abroad as well as the applications of actual peak regulation projects are discussed. Given the special working conditions faced by thermal storage technology in power grid peak regulation， four critical evaluation criteria， including safety， performance matching， reliability and economy that must be considered for thermal storage technology to participate in power grid peak regulation are proposed. The main business models， including energy management contracting （EMC）， two-part tariff （TPT）， and ancillary service market （ASM） for thermal storage to participate in power trading are explored. These models serve as valuable references for enabling the widespread commercial operation of thermal storage technology within the context of grid peak regulation.

Developing the proton exchange membrane (PEM) water electrolysis technology with flexibility in a wider load is an effective pathway to couple renewable energies with water electrolysis for hydrogen production and to achieve renewable energy consumption. This study first reviews scenarios of hydrogen production through the coupling of renewable electricity such as wind and photovoltaic power with fluctuating loads, and analyzes the fluctuation characteristics of renewable energy. Subsequently, it elaborates on the basic characteristics and research progress of water electrolysis for hydrogen production from three aspects: effect of fluctuating power on electrolysis cells, accelerated degradation of electrolysis components, and simulation methods for fluctuating power. Furthermore, the research and development directions of PEM electrolysis cell technology and PEM electrolysis for hydrogen production are explored. The current status and economic feasibility of wind-solar-coupled hydrogen production as well as the industrial application trends of hydrogen production under fluctuating power are clarified. Finally, we propose the following suggestions: (1) deepening the research of fundamental scientific issues and core components of electrolysis cells, (2) further reducing hydrogen production costs, and (3) optimizing the layout of wind-solar-coupled hydrogen production and institutional guarantee.

As a key component of energy storage technology， solid electric heat storage technology is adopted to convert valley electricity and various fluctuating electricity energy such as wind power and solar power into heat. The technology excels in environmental protection， efficiency， energy saving and safety and exhibits great potentials in regional heating renovation， deep peak regulating of the power grid and heat supply for industrial production. To this end， the principle of solid electric heat storage technology is introduced， and the research status and progress of solid heat storage materials and device structure are summarized. Moreover， bottleneck and key direction of the development of solid electric heat storage technology is discussed. Finally， the application status of the technology in different fields at home and abroad is sorted out and analyzed economically to provide ideas for promoting the development of the technology.

Objective To evaluate the in vitro antimicrobial activities of 16 common agents against Staphylococcus aureus, Enterococcus faecium and Enterococcus faecalis to provide data for the rational clinical use of drugs. Methods From 1st January to 31st December in 2019, Staphylococcus aureus, Enterococcus faecium and Enterococcus faecalis were collected from 24 hospitals in 18 provinces in China. Antimicrobial susceptibility testing was conducted by automated instrumentation method, and WHONET 5.6 software was used to calculate the drug sensitivity results of the strains to 16 antibacterial drugs including vancomycin, norvancomycin, fusidic acid and linezolid. Results A total of 255 strains of Staphylococcus aureus, 117 strains of Enterococcus faecalis and 114 strains of Enterococcus faecium were collected. Blood was the most common specimen type (22.2%, 108/486), followed by urine (18.9%, 82/486). According to the drug sensitivity results, vancomycin, norvancomycin, linezolid and teicoplanin maintained high activity to all strains. All Staphylococcus aureus and Enterococcus faecalis were sensitive to vancomycin and teicoplanin, while the resistance rates of vancomycin and teicoplanin against Enterococcus faecium were 4.4% and 2.6%, respectively. The resistance rates of linezolid against Staphylococcus aureus, Eenterococcus faecium and Eenterococcus fascalis were 0, 0 and 4.3%, respectively. Norvancomycin and vancomycin showed similar antibacterial activity against Staphylococcus aureus, Enterococcus faecium, and Enterococcus faecalis, with 50% minimum inhibitory concentration (MIC50) of 1 mg/L, and MIC90 of 2 mg/L. The isolation rate of methicillin-resistant Staphylococcus aureas(MRSA) was 35.7%, with 13.2% resistance to fusidic acid and 3.0% in methicillin-susceptible Staphylococcus aureas. Conclusions The activity of the 16 antimicro-bial drugs against Staphylococcus aureus, Enterococcus faecium and Enterococcus faecalis varied considerably. The data will provide reference for the rational use of antimicrobial drugs.

When high-speed trains enter tunnels, an initial compression wave is generated. As the compression wave propagates at the local speed of sound to the tunnel exit, it radiates into the surrounding environment, forming micro-pressure waves (MPWs). MPWs create sonic booms, resulting in significant environmental issues. The magnitude of the micro-pressure waves is directly proportional to the pressure gradient of the compression wave at the tunnel exit. The nonlinear effects of the initial compression wave during propagation lead to a significant increase in pressure gradient. Therefore, the propagation characteristics of the initial compression wave during the tunnel are the crucial factor affecting the amplitude of MPWs. Based on the one-dimensional compressible unsteady non-isentropic flow model and the improved generalized Riemann variable characteristic method, this paper researched the propagation and evolution characteristics of an initial compression wave generated when 400 km/h high-speed trains enter tunnels with three portal shapes: (no tunnel entrance hood (no hood), an oblique, enlarged tunnel entrance hood (type A), an enlarged equal-section non-uniform opening hole tunnel entrance hood (type B)). The results show that when the initial compression wave propagates inside very long tunnels, the pressure gradient of the compression wave exhibits a trend of initially increasing and then decreasing with the increase in propagation distance. When the pressure gradient of the compression wave reaches its maximum value, the corresponding propagation distance is the steepening critical distance. For no tunnel entrance hoods, type A tunnel entrance hoods, and type B tunnel entrance hoods, the steepening critical distances are 5 km, 6 km, and 16 km, respectively. The steepening critical distance shortens with increasing train speed. Steady friction and unsteady friction effects mainly affect the pressure amplitude and pressure gradient during compression wave propagation, respectively. At lower ambient temperatures, the nonlinear effects in compression wave propagation are significantly enhanced. The mitigation effects of type A tunnel entrance hoods and type B tunnel entrance hoods on pressure gradient reduction are mainly concentrated within 4 km and 12 km, respectively. It is necessary to determine the optimal matching relationship between the tunnel entrance hood and tunnel length based on the characteristics of compression wave propagation to ensure their mitigating performance is maximized.

Abstract The shape and internal structure of an atomic nucleus can change significantly with increasing excitation energy, angular momentum, or isospin asymmetry. As an example of this structural evolution, linear-chain configurations in carbon or heavier isotopes have been predicted for decades. Recent studies have found non-stability of this structure in 12C while evidenced its appearance in 16C. It is then necessary to investigate the linear-chain molecular structures in 14C to clarify the exact location on the nuclear chart where this structure begins to emerge, and thus to benchmark theoretical models. Here we show a cluster-decay experiment for 14C with all final particles coincidentally detected, allowing a high Q-value resolution, and thus a clear decay-path selection. Unambiguous spin-parity analyses are conducted, strongly evidencing the emergence of the π-bond linear-chain molecular rotational band in 14C. The present results encourage further studies on even longer chain configurations in heavier neutron-rich nuclei.

Osteoarthritis (OA) is a prevalent degenerative disease that impairs limb function, and its pathogenesis is closely related to inflammation. Sakuranetin (SK) is a cherry flavonoid phytoalexin with potent anti-inflammatory, anti-oxidant, and ant-ifungal properties. In recent studies, flavonoid and phytoalexin-related medicines have shown promise in the treatment of OA. However, the effects of SK on chondrocyte inflammation and the chondrogenesis process have remained unexplored, as have its functions in OA treatment. This study sought to confirm the therapeutic effects of SK in the OA rat model and reveal the potential mechanisms for protecting chondrocytes. The relevant mechanisms of SK were analyzed by network pharmacology analysis. Chondrocytes were subjected to IL-1β intervention to simulate an inflammatory environment and received SK treatment. Then, anabolism, catabolism, and inflammatory markers were detected by western blot, qPCR, elisa, and immunofluorescence. Chondrogenic ability was evaluated by micromass and 3D culture assays. The rats were treated with destabilization of the medial meniscus (DMM) surgery to establish an OA model and SK intra-articular injections subsequently. Histological staining, immunohistochemistry, and micro-CT were performed to analyze the structural and morphological changes of cartilage and subchondral bone. In chondrocytes, IL-1β treatment reduced chondrogenic ability, promoted catabolism, and exacerbated inflammation by triggering the PI3K/AKT/NF-κB pathway, whereas SK treatment partially rescued these negative effects. In vivo, SK treatment effectively alleviated the degeneration of cartilage and subchondral bone, thereby delaying the progression of OA. In summary, SK alleviates chondrocyte inflammation and promotes chondrogenesis by inhibiting the PI3K/AKT/NF-κB pathway, thereby improving OA progression.

Photosynthesis plays vital roles in plant growth and development. Stay-green (SGR) proteins are responsible for chlorophyll degradation and photosynthetic metabolism. To identify SGR family members and determine their potential functions in tea plants, we identified and cloned three SGR genes. Phylogenetic analysis revealed that the tea SGR homologs were classified into the SGR subfamily (named CsSGR1 and CsSGR2) and the SGRL subfamily (named CsSGRL). Cis-element analysis indicated that the promoters of CsSGR1, CsSGR2 and CsSGRL contained light-, phytohormone- and stress-related elements. Subcellular localization confirmed that CsSGR1 was localized in the chloroplast, while CsSGR2 and CsSGRL were localized in the chloroplast, membrane and nucleus. The RT-qPCR results showed that the three genes in the matures of albino tea cultivars were expressed higher than in the green tea cultivar. However, only CsSGR2-overexpressing tobacco leaves exhibited a yellowish phenotype and significantly lower Fv/Fm values. CsSGR1 and CsSGR2 exhibited similar expression patterns in different tissues after infection with the pathogen Colletotrichum camelliae, which was opposite to the pattern observed for CsSGRL. In addition, CsSGR1 was significantly induced in response to cold stress, SA, JA and ABA in C. camelliae. These findings identified valuable candidate genes for elucidating the mechanism of leaf albinism, stress response and phytohormone signaling in tea plants.

IntroductionEnterotoxigenic bacteria commonly excrete heat-labile enterotoxins (LT) as virulence factors that consist of one subunit A (LTA) and five B subunits (LTB). In fungi, there are a large number of genes encoding the homologs of LTA, but their biological roles remain largely unknown. MethodsIn this study, we identified 14 enterotoxin_A domain proteins in filamentous fungus B. bassiana in which five proteins were functionally characterized. ResultsFive proteins displayed diverse sub-cellular localizations but perform convergent functions in stress response, development, and virulence. The loss of five LTA genes resulted in significant reduction in conidial production, blastospore formation, and the increased sensitivity to oxidative and cell wall –perturbing stresses. The virulence of five disruptants was notably weakened as indicated by topical and intrahemocoel injection assays. Notably, the loss of these five proteins led to the significant changes in the carbohydrate profiles of cellular surface, which induced the enhanced host immune reactions of encapsulation and melanization. DiscussionThus, LTA proteins contribute to the fungus–host interaction via maintaining the carbohydrate profiles of cellular surface. This study expands our understanding of the enterotoxin_A domain proteins in fungal physiology and deepens mechanisms involved in the lifestyle of fungal insect pathogens.

In comparison to conventional actuated techniques, the utilization of a Flexible Biomimetic Artificial Muscle (FBAM) incorporating calcium alginate hydrogel, a type of responsive polymer, offers numerous benefits including reduced weight, lower actuated voltage requirements, enhanced specific energy density, and minimal acoustic disturbances, among others. This paper presents the fabrication of the FBAM through calcium alginate gelation utilizing six calcium salts with identical Ca2+ molar mass. Subsequently, the electrically actuated characteristics and electrochemical mechanism of the FBAM were investigated, employing four evaluation indexes: peak force density (D, mN/g), operating life (Tt, s), response speed (Vr, mN/g·s), and rise time (Ts, s). The findings of this study hold significant implications for the advancement of novel responsive polymers for artificial muscles. The findings of the study indicate that the electrically actuated characteristics of FBAM were most optimal when calcium alginate gelation with CaH8I2O4 was employed. This was evidenced by the D value of 22.807 mN/g, Tt value of 1066 s, Vr value of 98 s, and Ts value of 0.1046 mN/g·s. The sufficient degree of calcium alginate gelation between sodium alginate molecule and Ca2+ cations resulted in the formation of intermolecular bridges within the calcium alginate hydrogel, which consisted of polymer β-D-mannuronic and α-L-guluronic. Notably, this gelation process was found to be inversely related to the pH level in the aqueous solution of the electrically actuated membrane.

Rubber material is widely used in railway vehicles due to its superior damping performance. The testing methods, fatigue, and aging theories of rubber materials are of great significance to improve the design, manufacture, and application of rubber components for railway vehicles. This work systematically introduces the constitutive theory, mechanical testing standards, and testing methods of rubber materials. Then, the aging mechanism is described and the research progress of rubber fatigue properties is reviewed from the perspectives of fatigue crack initiation and fatigue crack propagation. Moreover, the reinforcement methods of rubber materials are presented. Finally, according to the working conditions of rubber components in railway vehicle, the technical difficulties and future research trends of fatigue characteristics analysis of rubber materials and components are pointed out.

Hydrolyzed tannin 1,2,6-tri-O-galloyl-β-D-glucopyranose (1,2,6-TGGP) possesses significant medicinal properties. However, little is known about its underlying molecular mechanisms. In this study, the levels of 1,2,6-TGGP in tea materials from different cultivars and leaf positions were compared. Additionally, one leaf and one bud sample from six tea cultivars with significant variations in 1,2,6-TGGP levels were analyzed using transcriptome high-throughput sequencing to identify the genes that are responsible for 1,2,6-TGGP accumulation. The sequencing results were mapped to the reference tea genome, revealing a total of 2735 differentially expressed genes (DEGs). This set included four UDP glycosyltransferase (UGTs) and six serine carboxypeptidases-like (SCPLs) genes. Among them, the upregulated SCPLs (CSS0032817) may directly participate in the acylation reaction of 1,2,6-TGGP. In addition, several classes of DEGs, including cytochrome P450, were significantly associated with the 1,2,6-TGGP content, which is potentially involved in their regulation. Overall, these results provide new insights into the molecular mechanism of 1,2,6-TGGP accumulation.

Leaf color variations in tea plants were widely considered due to their attractive phenotypes and characteristic flavors. The molecular mechanism of color formation was extensively investigated. But few studies focused on the transformation process of leaf color change. In this study, four strains of ‘Baijiguan’ F1 half-sib generation with similar genetic backgrounds but different colors were used as materials, including Green (G), Yellow-Green (Y-G), Yellow (Y), and Yellow-Red (Y-R). The results of broadly targeted metabolomics showed that 47 metabolites were differentially accumulated in etiolated leaves (Y-G, Y, and Y-R) as compared with G. Among them, lipids were the main downregulated primary metabolites in etiolated leaves, which were closely linked with the thylakoid membrane and chloroplast structure. Flavones and flavonols were the dominant upregulated secondary metabolites in etiolated leaves, which might be a repair strategy for reducing the negative effects of dysfunctional chloroplasts. Further integrated analysis with the transcriptome indicated different variation mechanisms of leaf phenotype in Y-G, Y, and Y-R. The leaf color formation of Y-G and Y was largely determined by the increased content of eriodictyol-7-O-neohesperidoside and the enhanced activities of its modification process, while the color formation of Y-R depended on the increased contents of apigenin derivates and the vigorous processes of their transportation and transcription factor regulation. The key candidate genes, including UDPG, HCT, CsGSTF1, AN1/CsMYB75, and bHLH62, might play important roles in the flavonoid pathway.

Abstract Background Tea plant breeding or cultivation mainly involves propagation via cuttings, which not only ensures the inheritance of the excellent characteristics of the mother plant but also facilitates mechanized management. The formation of adventitious root (AR) determines the success of cutting-based propagation, and auxin is an essential factor involved in this process. To understand the molecular mechanism underlying AR formation in nodal tea cuttings, transcriptome and endogenous hormone analysis was performed on the stem bases of red (mature)- and green (immature)-stem cuttings of ‘Echa 1 hao’ tea plant as affected by a pulse treatment with naphthalene acetic acid (NAA). Results In this study, NAA significantly promoted AR formation in both red- and green-stem cuttings but slightly reduced callus formation. External application of NAA reduced the levels of endogenous indole-3-acetic acid (IAA) and cytokinin (TZR, trans-zeatin riboside). The number of DEGs (NAA vs. CK) identified in the green-stem cuttings was significantly higher than that in the red-stem cuttings, which corresponded to a higher rooting rate of green-stem cuttings under the NAA treatment. A total of 82 common DEGs were identified as being hormone-related and involved in the auxin, cytokinin, abscisic acid, ethylene, salicylic acid, brassinosteroid, and jasmonic acid pathways. The negative regulation of NAA-induced IAA and GH3 genes may explain the decrease of endogenous IAA. NAA reduced endogenous cytokinin levels and further downregulated the expression of cytokinin signalling-related genes. By the use of weighted gene co-expression network analysis (WGCNA), several hub genes, including three [cellulose synthase (CSLD2), SHAVEN3-like 1 (SVL1), SMALL AUXIN UP RNA (SAUR21)] that are highly related to root development in other crops, were identified that might play important roles in AR formation in tea cuttings. Conclusions NAA promotes the formation of AR of tea cuttings in coordination with endogenous hormones. The most important endogenous AR inductor, IAA, was reduced in response to NAA. DEGs potentially involved in NAA-mediated AR formation of tea plant stem cuttings were identified via comparative transcriptome analysis. Several hub genes, such as CSLD2, SVL1 and SAUR21, were identified that might play important roles in AR formation in tea cuttings.

Abstract Low‐temperature composite insulation is commonly applied in high‐temperature superconducting apparatus while partial discharge (PD) is found to be an important indicator to reveal insulation statues. In order to extract feature parameters of PD signals more effectively, a method combined variational mode decomposition with multi‐scale entropy and image feature is proposed. Based on the simulated test platform, original and noisy signals of three typical PD defects were obtained and decomposed. Accordingly, relative moments and grayscale co‐occurrence matrix were employed for feature extraction by K‐modal component diagram. Afterwards, new PD feature vectors were obtained by dimension reduction. Finally, effectiveness of different feature extraction methods was evaluated by pattern recognition based on support vector machine and K‐nearest neighbour. Result shows that the proposed feature extraction method has a higher recognition rate by comparison and is robust in processing noisy signals.

Abstract Extracellular vesicle (EV)-mediated transfer of long non-coding RNAs (lncRNAs) has been reported to regulate chemoresistance in various cancers. We herein investigate the therapeutic potential of bioinformatically identified HOTAIR transferred by serum-derived EVs (serum-EVs) in temozolomide (TMZ) resistance of glioblastoma (GBM) and the downstream mechanisms. EVs were isolated from the serum of GBM patients. Expression of HOTAIR was examined in the clinical tissue samples and serum-EVs of GBM patients. The downstream miRNAs of HOTAIR and its target genes were predicted in silico. The effects of the HOTAIR transmitted by serum-EVs in malignant phenotypes, tumor growth, and TMZ resistance were assessed in vitro and in vivo. HOTAIR expression was upregulated in clinical tissues, cells, and serum-EVs of GBM. Co-culture data showed that GBM-serum-EVs facilitated GBM cell proliferative and invasive phenotypes and TMZ resistance by elevating HOTAIR. In GBM cells, HOTAIR competitively bound to miR-526b-3p and weakened miR-526b-3p’s binding ability to EVA1, thus increasing the expression of EVA1. Furthermore, HOTAIR carried by serum-EVs promoted tumor growth and TMZ resistance in vivo by suppressing miR-526b-3p-mediated EVA1 inhibition. GBM-serum-EV-enclosed HOTAIR may augment GBM progression and chemoresistance through miR-526b-3p downregulation and EVA1 upregulation. These results provide a strategy to reduce TMZ resistance in GBM treatment.

The research on the activity of selenium (Se)-enriched agricultural products is receiving increasing attention since Se was recognized for its antioxidant activities and for its enhancement of immunity in trace elements. In this study, antioxidant Se-containing peptides, namely, Se-TAPepI-1 and Se-TAPepI-2, were optimally separated and prepared from Se-enriched tea protein hydrolysates by ultrafiltration and Sephadex G-25 purification, and subsequently, their physicochemical properties, oligopeptide sequence, and potential antioxidant mechanism were analyzed. Through the optimization of enzymatic hydrolysis conditions, the Se-enriched tea protein hydrolyzed by papain exhibited a better free radical scavenging activity. After separation and purification of hydrolysates, the two peptide fractions obtained showed significant differences in selenium content, amino acid composition, apparent morphology, peptide sequence, and free radical scavenging activity. Therein, two peptides from Se-TAPepI-1 included LPMFG (563.27 Da) and YPQSFIR (909.47 Da), and three peptides from Se-TAPepI-2 included GVNVPYK (775.42 Da), KGGPGG (552.24 Da), and GDEPPIVK (853.45 Da). Se-TAPepI-1 and Se-TAPepI-2 could ameliorate the cell peroxidation damage and inflammation by regulating NRF2/ARE pathway expression. Comparably, Se-TAPepI-1 showed a better regulatory effect than Se-TAPepI-2 due to their higher Se content, typical amino acid composition and sequence, higher surface roughness, and a looser arrangement in their apparent morphology. These results expanded the functional activities of tea peptide and provided the theoretical basis for the development of Se-containing peptides from Se-enriched tea as a potential natural source of antioxidant dietary supplements.

Caffeine is a characteristic secondary metabolite in tea plants. It confers tea beverage with unique flavor and excitation effect on human body. The pathway of caffeine biosynthesis has been generally established, but the mechanism of caffeine transport remains unclear. Here, eight members of purine permeases (PUPs) were identified in tea plants. They had diverse expression patterns in different tissues, suggesting their broad roles in caffeine metabolism. In this study, F1 strains of "Longjing43" ♂ × "Baihaozao" ♀ and different tea cultivars were used as materials to explore the correlation between caffeine content and gene expression. The heterologous expression systems of yeast and Arabidopsis were applied to explore the function of CsPUPs. Correlation analysis showed that the expressions of CsPUP1, CsPUP3.1, and CsPUP10.1 were significantly negatively correlated with caffeine content in tea leaves of eight strains and six cultivars. Furthermore, subcellular localization revealed that the three CsPUPs were not only located in plasma membrane but also widely distributed as circular organelles in cells. Functional complementation assays in yeast showed that the three CsPUPs could partly or completely rescue the defective function of fcy2 mutant in caffeine transport. Among them, transgenic yeast of CsPUP10.1 exhibited the strongest transport capacity for caffeine. Consistent phenotypes and functions were further identified in the CsPUP10.1-over-expression Arabidopsis lines. Taken together, it suggested that CsPUPs were involved in caffeine transport in tea plants. Potential roles of CsPUPs in the intracellular transport of caffeine among different subcellular organelles were proposed. This study provides a theoretical basis for further research on the PUP genes and new insights for caffeine metabolism in tea plants.

Vehicle classification is getting important especially in security systems, surveillance, transportation congestion reduction, and accident prevention. However, it is difficult to classify the traffic objects due to the poor quality of images from videos. Hence, image processing techniques are applied to increase the accuracy of the result. The aim of this study is to propose a vehicle classification scheme where YOLO v5 algorithm and Faster R-CNN algorithm are being implemented separately into vehicle classification, followed by comparison of result between these two algorithms. In this study, vehicles are classified into five classes, namely motorcycle, car, van, bus and lorry. The labeled dataset is being split into training set and validation set and then trained under algorithm YOLO v5 and Faster R-CNN separately. Experimental results show that YOLO v5 performs better with the mean average Precision, Precision, and Recall rate up to 0.91, 0.81, and 0.86, respectively

Severe plastic deformation (SPD) with deformation strain significantly larger than one is effective to strengthen metals and alloys. However, such a method will not work for Mg alloys due to their low deformability. In view of this problem, we propose a new alloy strengthening scheme of low-strain deformation, which is to introduce massive twins and stacking faults in coarse grain interior by rotary swaging (RS) at room temperature. By using this scheme, we successfully prepared an ultra-light bulk Mg-Li alloy with new strength record.

Abstract Total hip arthroplasty and total knee arthroplasty are extensively used for the treatment of the end-stage degenerative joint diseases. Currently, periprosthetic bone loss is still the major cause of aseptic loosening, resulting in implant failures. Previous literature introduced some widely accepted protocols for the prevention and treatment of periprosthetic bone loss, but no guideline has been proposed. Denosumab, a human monoclonal immunoglobulin G2 (IgG2) antibody, can inhibit bone resorption by binding to the receptor activator of nuclear factor kappa-B ligand (RANKL). This article reviews the present findings and evidence concerning the effect of denosumab on the periprosthetic bone loss after total hip arthroplasty and total knee arthroplasty. Overall, the current evidence suggests that denosumab is a promising agent for the treatment of periprosthetic bone loss.

Abstract Background Alanine decarboxylase (AlaDC), specifically present in tea plants, is crucial for theanine biosynthesis. Serine decarboxylase (SDC), found in many plants, is a protein most closely related to AlaDC. To investigate whether the new gene AlaDC originate from gene SDC and to determine the biochemical properties of the two proteins from Camellia sinensis, the sequences of CsAlaDC and CsSDC were analyzed and the two proteins were over-expressed, purified, and characterized. Results The results showed that exon-intron structures of AlaDC and SDC were quite similar and the protein sequences, encoded by the two genes, shared a high similarity of 85.1%, revealing that new gene AlaDC originated from SDC by gene duplication. CsAlaDC and CsSDC catalyzed the decarboxylation of alanine and serine, respectively. CsAlaDC and CsSDC exhibited the optimal activities at 45 °C (pH 8.0) and 40 °C (pH 7.0), respectively. CsAlaDC was stable under 30 °C (pH 7.0) and CsSDC was stable under 40 °C (pH 6.0–8.0). The activities of the two enzymes were greatly enhanced by the presence of pyridoxal-5′-phosphate. The specific activity of CsSDC (30,488 IU/mg) was 8.8-fold higher than that of CsAlaDC (3467 IU/mg). Conclusions Comparing to CsAlaDC, its ancestral enzyme CsSDC exhibited a higher specific activity and a better thermal and pH stability, indicating that CsSDC acquired the optimized function after a longer evolutionary period. The biochemical properties of CsAlaDC might offer reference for theanine industrial production.

As an important research direction in image and video processing, set-based video recognition requires speed and accuracy. However, the existing static modeling methods focus on computational speed but ignore accuracy, whereas the dynamic modeling methods are higher-accuracy but ignore the computational speed. Combining these two types of methods to obtain fast and accurate recognition results remains a challenging problem. Motivated by this, in this study, a novel Manifolds-based Low-Rank Dictionary Pair Learning (MbLRDPL) method was developed for a set-based video recognition/image set classification task. Specifically, each video or image set was first modeled as a covariance matrix or linear subspace, which can be seen as a point on a Riemannian manifold. Second, the proposed MbLRDPL learned discriminative class-specific synthesis and analysis dictionaries by clearly imposing the nuclear norm on the synthesis dictionaries. The experimental results show that our method achieved the best classification accuracy (100%, 72.16%, 95%) on three datasets with the fastest computing time, reducing the errors of state-of-the-art methods (JMLC, DML, CEBSR) by 0.96–75.69%.

Autophagy is a conserved mechanism for the turnover of intracellular components. Among the ‘core’ autophagy-related genes (ATGs), the cysteine protease Atg4 plays an important role in the activation of Atg8 by exposing the glycine residue at its extreme carboxyl terminus. In the insect fungal pathogen Beauveria bassiana, a yeast ortholog of Atg4 was identified and functionally analyzed. Ablation of the BbATG4 gene blocks the autophagic process during fungal growth under aerial and submerged conditions. Gene loss did not affect fungal radial growth on various nutrients, but ΔBbatg4 exhibited an impaired ability to accumulate biomass. The mutant displayed increased sensitivity to stress caused by menadione and hydrogen peroxide. ΔBbatg4 generated abnormal conidiophores with reduced production of conidia. Additionally, fungal dimorphism was significantly attenuated in gene disruption mutants. Disruption of BbATG4 resulted in significantly weakened virulence in topical and intrahemocoel injection assays. Our study indicates that BbAtg4 contributes to the lifecycle of B. bassiana via its autophagic roles.

Background: Although recent research suggests that alterations in gut microbiota and metabolites play a critical role in the pathophysiology of immunoglobulin A nephropathy (IgAN), the causal relationship between specific intestinal flora and metabolites and the risk of IgAN remains unclear. Method: This study employed Mendelian randomization (MR) to investigate the causal association between gut microbiota and IgAN. To explore potential associations between gut microbiota and various outcomes, four MR methods were applied: inverse variance weighted (IVW), MR-Egger, weighted median, and weighted mode. If the results of the four methods are inconclusive, we prefer the IVW as the primary outcome. Additionally, MR-Egger, MR-PRESSO-Global, and Cochrane’s Q tests were used to detect heterogeneity and pleiotropy. The stability of MR findings was assessed using the leave-one-out approach, and the strength of the causal relationship between exposure and outcome was tested using Bonferroni correction. Additional clinical samples were utilized to validate the results of Mendelian randomization, and the outcomes were visualized through an ROC curve, confusion matrix, and correlation analysis. Result: This study examined a total of 15 metabolites and 211 microorganisms. Among them, eight bacteria and one metabolite were found to be associated with the risk of IgAN (p < 0.05). The Bonferroni-corrected test reveals that only Class. Actinobacteria (OR: 1.20, 95% CI: 1.07–1.36, p = 0.0029) have a significant causal relationship with IgAN. According to Cochrane’s Q test, there is no substantial heterogeneity across different single-nucleotide polymorphisms (p > 0.05). Furthermore, MR-Egger and MR-PRESSO-Global tests (p > 0.05) showed no evidence of pleiotropy. No reverse causal association was found between the risk of IgAN and microbiota or metabolites (p > 0.05). Clinical specimens demonstrated the effectiveness and accuracy of Actinobacteria in distinguishing IgAN patients from those with other glomerular diseases (AUC = 0.9, 95% CI: 0.78–1.00). Additionally, our correlation analysis revealed a potential association between Actinobacteria abundance and increased albuminuria (r = 0.85) and poorer prognosis in IgAN patients (p = 0.01). Conclusion: Through MR analysis, we established a causal link between Actinobacteria and the incidence of IgAN. Moreover, clinical validation using fecal samples indicated that Actinobacteria might be associated with the onset and poorer prognosis of IgAN. This finding could provide valuable biomarkers for early, noninvasive detection of the disease and potential therapeutic targets in IgAN.

Fenvalerate has the advantages of a broad insecticidal spectrum, high efficiency, low toxicity and low cost, and it is widely used in agriculture, especially in tea, resulting in the accumulation of fenvalerate residues in tea and the environment, posing a serious threat to human health. Therefore, the timely monitoring of fenvalerate residue dynamics is vital for ensuring the health of humans and the ecological environment, and it is necessary for establishing a fast, reliable, accurate and on-site method for detecting fenvalerate residues. Based on the methods of immunology, biochemistry and molecular biology, mammalian spleen cells, myeloma cells and mice were used as experimental materials to establish a rapid detection method of an enzyme-linked immunosorbent assay to detect the residues of fenvalerate in dark tea. Three cell lines—1B6, 2A11 and 5G2—that can stably secrete fenvalerate antibodies were obtained by McAb technology, and their sensitivities (IC50) were 36.6 ng/mL, 24.3 ng/mL and 21.7 ng/mL, respectively. The cross-reaction rates of the pyrethroid structural analogs were all below 0.6%. Six dark teas were used to detect the practical application of fenvalerate monoclonal antibodies. The sensitivity IC50 of the anti-fenvalerate McAb in PBS with 30% methanol is 29.12 ng/mL. Furthermore, a latex microsphere immunochromatographic test strip with an LOD of 10.0 ng/mL and an LDR of 18.9–357 ng/mL was preliminarily developed. A specific and sensitive monoclonal antibody for fenvalerate was successfully prepared and applied to detect fenvalerate in dark teas (Pu‘er tea, Liupao tea, Fu Brick tea, Qingzhuan tea, Enshi dark tea and selenium-enriched Enshi dark tea). A latex microsphere immunochromatographic test strip was developed for the preparation of rapid detection test strips of fenvalerate.

Osteoarthritis (OA), the most common chronic inflammatory joint disease, is characterized by progressive cartilage degeneration, subchondral bone sclerosis, synovitis, and osteophyte formation. Metformin, a hypoglycemic agent used in the treatment of type 2 diabetes, has been evidenced to have anti-inflammatory properties to treat OA. It hampers the M1 polarization of synovial sublining macrophages, which promotes synovitis and exacerbates OA, thus lessening cartilage loss. In this study, metformin prevented the pro-inflammatory cytokines secreted by M1 macrophages, suppressed the inflammatory response of chondrocytes cultured with conditional medium (CM) from M1 macrophages, and mitigated the migration of M1 macrophages induced by interleukin-1ß (IL-1ß)-treated chondrocytes in vitro. In the meantime, metformin reduced the invasion of M1 macrophages in synovial regions brought about by the destabilization of medial meniscus (DMM) surgery in mice, and alleviated cartilage degeneration. Mechanistically, metformin regulated PI3K/AKT and downstream pathways in M1 macrophages. Overall, we demonstrated the therapeutic potential of metformin targeting synovial M1 macrophages in OA.

A new expansion-continuous shear deformation technique was used to prepare the 6061 aluminum alloy sheet. The extrusion deformation behavior of 6061 aluminum alloy was discussed by the finite element simulation. The microstructure and mechanical properties of 6061 aluminum alloy were studied via the testing and analysis tools of electron back-scatter diffraction,tensile test and scanning electron microscope. The results show that the expansion-continuous shear deformation can coordinate the stability of metal flow velocity during the deformation process of the aluminum alloy. The severe plastic deformation can produce a large and uniform equivalent strain in the extrusion billet, which can effectively trigger continuous dynamic recrystallization, moreover, the orientation distribution of fine grains is scattered, which can effectively refine the grain and weaken the texture. After extrusion, 6061 aluminum alloy presents excellent mechanical properties, and the tensile strength and the elongation can reach up to 380 MPa and 21%, respectively. A large number of dimples emerge in the tensile fracture, which can be attributed to ductile fracture.

Due to the insufficient slip systems, Mg and its alloys exhibit poor ductility during plastic deformation at room temperature. To solve this problem, alloying is considered as a most effective method to improve the ductility of Mg alloys, which attracts wide attentions of industries. However, it is still a challenge to understand the ductilization mechanism, because of the complicated alloying elements and their interactions with Mg matrix. In this work, pure Mg and Mg-Y alloys were comparatively studied to investigate the effect of Y addition on microstructure evolution and mechanical properties. A huge increase of uniform elongation, from 5.3% to 20.7%, was achieved via only 3 wt% addition of yttrium. TEM results revealed that the only activated slip system in pure Mg was basal slip, led to its poor ductility at room temperature. In contrast, a large number of stacking faults and non-basal dislocations with component were observed in the deformed Mg-Y alloy, which was proposed as the main reason for significant improvement of strain hardening and ductility. High resolution TEM indicated that most of the stacking faults were I1 and I2 intrinsic faults, which played a critical role in improving the ductility of Mg-Y alloy. Addition of Y into Mg alloy decreased the stacking fault energy, which induced high density stacking faults in the grain interior.

Selenium-enriched mung bean (Se-MB) is a combination of mung bean (MB) and selenium (Se), which have a variety of potential biological activities. However, little is known about the skincare activity of Se-MB. The chemical composition of Se-MB fermentation broth (Se-MBFB) was analyzed to investigate the whitening, moisturizing, and anti-aging activities of Se-MBFB. The tyrosinase inhibition, anti-melanogenic in melanocytes (B16F10 cells), and moisturizing effect in human dermal fibroblasts (HDFs) were analyzed. Besides, the free radical scavenging activity of Se-MBFB was assessed in vitro. To verify the in vivo effects and the potential of practical applications of Se-MBFB, a clinical trial was conducted on the participants: 31 Chinese women aged 25–60 years, with no pigmentation disorder, no illness, no history of hypersensitivity reaction, and no use of skincare product on the face. The participants used an Se-MBFB masque for 15-20 min after cleaning the face. The measurement points were Week 0, 2, and 4 (W0, W2, and W4) after using the masque, and target sites were cheek and canthus. The following parameters were recorded on the target sites at each visit: melanin index, skin color, cuticle moisture content, transepidermal water loss, and crow's feet. The results demonstrated that Se-MBFB was rich in polyphenols, peptides, and γ-aminobutyric acid (GABA), displayed significant free radical scavenging and tyrosinase inhibiting activities, decreased the synthesis of melanin, and upregulated the aquaporin-3 (AQP3) expression. The test of the Se-MBFB mask showed that after 4 weeks of using the Se-MBFB facemask, the faces of the participants became whiter with reduced wrinkles and increased moisture content. Se-MB possessed the excellent whitening, moisturizing, and antioxidant efficacy, which could lay a scientific foundation for utilization and development of skincare products of Se-MB and its related industrial cosmetics products.

Tea is an important beverage crop with a large and heterozygous genome. Here, the authors assemble the genome of the cultivar Longjing 43 and conduct a population genetics study to reveal divergent selection for disease resistance and flavor between the two variety groups.

Glioma is emerging as an aggressive type of glioma characterized by invasive growth pattern and dismal oncologic outcomes. microRNAs (miRNAs) have been attracting research attention in tumorigenesis. Herein, the aim of the current investigation was to explore the functional role of mesenchymal stem cells (MSCs)-derived extracellular vesicles (EVs) containing miR-503 in glioma. The glioma tissues and corresponding normal brain tissues were collected from patients with glioma, followed by quantification of miR-503, kinesin family member 5A (KIF5A) and interleukin-7 (IL-7). EVs were isolated from bone marrow MSCs and identified by transmission electron microscope and nanoparticle tracking analysis. EVs from miR-503 mimic-transfected MSCs, miR-503 agomir,, oe-KIF5A, or sh-IL-7 was delivered into glioma cells to determine their effects on biological behaviors of glioma and T cells as well as the release of immunosuppressive factors. Lastly, a mouse model of glioma was developed to validate the function in vivo. miR-503 was expressed at a high level in glioma tissues while KIF5A was poorly expressed and targeted by miR-503. Furthermore, miR-503 loaded in MSC-EVs or upregulated miR-503 was demonstrated to facilitate glioma cell proliferation, migration and invasion accompanied by promoted release of immunosuppressive factors. Effects of overexpressed KIF5A on T cell behavior modulation were dependent on the IL-7 signaling pathway. Such results were reproduced in mice with glioma. Collectively, the discovery of miR-503 incorporated in MSC-EVs being a regulator that controls immune escape in glioma provides a novel molecular insight that holds promises to develop therapeutic strategies against glioma.

Atmospheric pollution has always been an intractable regional problem that requires the cooperation of various cities and departments within a city cluster to achieve good results. As one of the key urban agglomerations in China, the Ha-Chang urban agglomeration is also plagued by the problem of air pollution. With the help of institutional collective action theory and social network analysis, we analyze the current situation of air pollution cooperation in the Ha-Chang urban agglomeration from two perspectives: inter-city cooperation network and inter-governmental cooperation network, and put forward suggestions for improving the strategic height and precision of treatment by calculating indicators such as network density and network centrality.

Tea plants are widely grown all over the world because they are an important economic crop. The purity and authenticity of tea varieties are frequent problems in the conservation and promotion of germplasm resources in recent years, which has brought considerable inconvenience and uncertainty to the selection of parental lines for breeding and the research and cultivation of superior varieties. However, the development of core SNP markers can quickly and accurately identify the germplasm, which plays an important role in germplasm identification and the genetic relationship analysis of tea plants. In this study, based on 179,970 SNP loci from the whole genome of the tea plant, all of 142 cultivars were clearly divided into three groups: Assam type (CSA), Chinese type (CSS), and transitional type. Most CSA cultivars are from Yunnan Province, which confirms that Yunnan Province is the primary center of CSA origin and domestication. Most CSS cultivars are distributed in east China; therefore, we deduced that east China (mainly Zhejiang and Fujian provinces) is most likely the area of origin and domestication of CSS. Moreover, 45 core markers were screened using strict criteria to 179,970 SNP loci, and we analyzed 117 well-Known tea cultivars in China with 45 core SNP markers. The results were as follows: (1) In total, 117 tea cultivars were distinguished by eight markers, which were selected to construct the DNA fingerprint, and the remaining markers were used as standby markers for germplasm identification. (2) Ten pairs of parent and offspring relationships were confirmed or identified, and among them, seven pairs were well-established pedigree relationships; the other three pairs were newly identified. In this study, the east of China (mainly Zhejiang and Fujian provinces) is most likely the area of origin and domestication of CSS. The 45 core SNP markers were developed, which provide a scientific basis at the molecular level to identify the superior tea germplasm, undertake genetic relationship analysis, and benefit subsequent breeding work.

The incidence of degenerative spinal diseases, such as cervical spondylosis and thoracic and lumbar disc herniation, is increasing. These health problems have adversely affected human life and work. Surgical intervention is effective when intervertebral disc degeneration (IDD) causes nerve compression and/or severely limits daily activity. Early IDD patients generally do not require surgery. However, there is no effective method of impeding IDD progression. Thus, novel approaches to alleviating IDD deterioration are urgently required. Cystathionine-γ-lyase (CSE) and E-selectin (CD62E) are vital factors regulating vascular function and inflammation. However, their effects on IDD and vascular invasion in intervertebral discs (IVDs) are pending further exploration. Here, bioinformatics and human nucleus pulposus (NP) tissues analyses revealed that CSE was significantly downregulated and CD62E was upregulated in the NP tissues of IDD patients. We demonstrated that CSE overexpression, CD62E downregulation, and NF-κB (P65) inhibition mitigate inflammation and recover metabolic function in NP cells. Similarly, CSE attenuated vascular invasion induced by inflammatory irritation. Using a rat IDD model, we showed that CSE improved degeneration, inflammation, and microvascular invasion in NP tissue, whereas CD62E had the opposite effect. Taken together, our results indicated that the CSE/CD62E pathway could effectively improve the inflammatory environment and vascular invasion in IVD. Hence, the findings of this study propose a promising and valuable strategy for the treatment of patients with early IDD as well as postoperative adjuvant therapy in patients with severe IDD.

Plant acyl-CoA dominated acyltransferases (named BAHD) comprise a large appointed protein superfamily and play varied roles in plant secondary metabolism like synthesis of modified anthocyanins, flavonoids, volatile esters, etc. Tea (Camellia sinensis) is an important non-alcoholic medicinal and fragrancy plant synthesizing different secondary metabolites, including flavonoids. In the tea (C.A sinensis) cultivar Longjing 43 (LJ43), eight samples were performed into three groups for transcriptome analysis under three biological replications. Among the BAHD acyltransferase genes in tea cultivars, the expression of TEA031065 was highest in buds and young leaves following the RNA sequencing data, which was coincident with the tissue rich in catechins and other flavonoids. We then transformed this gene into wild-type Arabidopsis as an over-expression (OX) line 1 and line 2 in ½ MS media to verify its function. In the wild types (WT), the primary root length, number of secondary roots, and total root weight were significantly higher at 24%, 15%, and 53.92%, respectively, compared to the transgenic lines (OX1 and OX2). By contrast, the leaves displayed larger rosettes (21.58%), with higher total leaf weight (32.64%) in the transgenic lines than in the wild type (WT). This result is consistent with DCR mutant At5g23940 gene in Arabidopsis thaliana. Here, anthocyanin content in transgenic lines was also increased (21.65%) as compared to WT. According to the RNA sequencing data, a total of 22 growth regulatory genes and 31 structural genes with TFs (transcription factors) that are correlative with plant growth and anthocyanin accumulation were identified to be differentially expressed in the transgenic lines. It was found that some key genes involved in IAA (Auxin) and GA (Gibberellin) biosynthesis were downregulated in the transgenic lines, which might be correlated with the phenotype changes in roots. Moreover, the upregulation of plant growth regulation genes, such as UGT73C4 (zeatin), ARR15, GH3.5, ETR2, ERS2, APH4, and SAG113 might be responsible for massive leaf growth. In addition, transgenic lines shown high anthocyanin accumulation due to the upregulation of the (1) 3AT1 and (3) GSTF, particularly, GSTF12 genes in the flavonoid biosynthesis pathway. However, the TFs such as, CCoAMT, bHLH, WRKY, CYP, and other MYBs were also significantly upregulated in transgenic lines, which increased the content of anthocyanins in A. thaliana seedlings. In conclusion, a BAHD acyltransferase (TEA031065) was identified, which might play a vital role in tea growth and secondary metabolites regulation. This study increases our knowledge concerning the combined functionality of the tea BAHD acyltransferase gene (TEA031065).

The accumulation of macrophages in degenerated discs is a common phenomenon. However, the roles and mechanisms of M2a macrophages in intervertebral disc degeneration (IDD) have not been illuminated. This study investigated the expression of the M2a macrophage marker (CD206) in human and rat intervertebral disc tissues by immunohistochemistry. To explore the roles of M2a macrophages in IDD, nucleus pulposus (NP) cells were co-cultured with M2a macrophages in vitro. To clarify whether the CHI3L1 protein mediates the effect of M2a macrophages on NP cells, siRNA was used to knock down CHI3L1 transcription. To elucidate the underlying mechanisms, NP cells were incubated with recombinant CHI3L1 proteins, then subjected to western blotting analysis of the IL-13Rα2 receptor and MAPK pathway. CD206-positive cells were detected in degenerated human and rat intervertebral disc tissues. Notably, M2a macrophages promoted the expression of catabolism genes (MMP-3 and MMP-9) and suppressed the expression of anabolism genes (aggrecan and collagen II) in NP cells. These effects were abrogated by CHI3L1 knockdown in M2a macrophages. Exposure to recombinant CHI3L1 promoted an extracellular matrix metabolic imbalance in NP cells via the IL-13Rα2 receptor, along with activation of the ERK and JNK MAPK signaling pathways. This study elucidated the roles of M2a macrophages in IDD and identified potential mechanisms for these effects.

NRT1/PTR FAMILY (NPF) genes are characterized as nitrate and peptide transporters that played important roles in various substrates transport in plants. However, little is known about the NPF gene in tea plants. Here, a total of 109 CsNPF members were identified from the tea plant genome, and divided into 8 groups according to their sequence characteristics and phylogenetic relationship. Gene structure and conserved motif analysis supported the evolutionary conservation of CsNPFs. Many hormone and stress response cis-acting elements and transcription factor binding sites were found in CsNPF promoters. Syntenic analysis suggested that multiple duplication types contributed to the expansion of NPF gene family in tea plants. Selection pressure analysis showed that CsNPF genes experienced strong purifying selective during the evolution process. The distribution of NPF family genes revealed that 8 NPF subfamilies were formed before the divergence of eudicots and monocots. Transcriptome analysis showed that CsNPFs were expressed differently in different tissues of the tea plant. The expression of 20 CsNPF genes at different nitrate concentrations was analyzed, and most of those genes responded to nitrate resupply. Subcellular localization showed that both CsNPF2.3 and CsNPF6.1 were localized in the plasma membrane, which was consistent with the characteristics of transmembrane proteins involved in NO3- transport. This study provides a theoretical basis for further investigating the evolution and function of NPF genes.

BackgroundMaternal solvent exposure has been suspected to increase offspring cancer risk. The study aimed to evaluate the associations between maternal residential exposure to solvents from industrial pollution during pregnancy and childhood cancer.MethodsThe present study included 15,744 cancer cases (aged 0-19 years at diagnosis) identified from California Cancer Registry and 283,141 controls randomly selected from California Birth Registry (20:1 frequency-matched by birth year: 1998-2016). We examined industrial releases of tetrachloroethylene and 1,1,1-trichloroethane within 3 km of the birth address, while we used a 5 km buffer for carbon disulfide. We calculated the total exposure from all linked Toxic Release Inventory sites during each index pregnancy and assigned "ever/never" and "high/low exposed/unexposed" exposure, using median values. We performed quadratic decay models to estimate cancer risks associated with maternal solvent exposure in pregnancy.Results1,1,1-Trichloroethane was associated with rhabdomyosarcoma (adjusted Odds Ratio (aOR): 1.96; 95% Confidence Interval (CI): 1.16, 3.32) in the "ever exposed" group. Ever exposure to carbon disulfide was associated with increased risks of medulloblastoma (OR = 1.85, 95% CI 1.01, 3.40) and ependymoma (OR = 1.63, 95% CI 0.97, 2.74).ConclusionsOverall, our findings suggested maternal residential exposure to solvents from industrial sources might be associated with elevated childhood cancer risks.

Abstract Background Flavonoids are important components that confer upon tea plants a unique flavour and health functions. However, the traditional breeding method for selecting a cultivar with a high or unique flavonoid content is time consuming and labour intensive. High-density genetic map construction associated with quantitative trait locus (QTL) mapping provides an effective way to facilitate trait improvement in plant breeding. In this study, an F1 population (LJ43×BHZ) was genotyped using 2b-restriction site-associated DNA (2b-RAD) sequencing to obtain massive single nucleotide polymorphism (SNP) markers to construct a high-density genetic map for a tea plant. Furthermore, QTLs related to flavonoids were identified using our new genetic map. Results A total of 13,446 polymorphic SNP markers were developed using 2b-RAD sequencing, and 4,463 of these markers were available for constructing the genetic linkage map. A 1,678.52-cM high-density map at an average interval of 0.40 cM with 4,217 markers, including 427 frameset simple sequence repeats (SSRs) and 3,800 novel SNPs, mapped into 15 linkage groups was successfully constructed. After QTL analysis, a total of 27 QTLs related to flavonoids or caffeine content (CAF) were mapped to 8 different linkage groups, LG01, LG03, LG06, LG08, LG10, LG11, LG12, and LG13, with an LOD from 3.14 to 39.54, constituting 7.5% to 42.8% of the phenotypic variation. Conclusions To our knowledge, the highest density genetic map ever reported was constructed since the largest mapping population of tea plants was adopted in present study. Moreover, novel QTLs related to flavonoids and CAF were identified based on the new high-density genetic map. In addition, two markers were located in candidate genes that may be involved in flavonoid metabolism. The present study provides valuable information for gene discovery, marker-assisted selection breeding and map-based cloning for functional genes that are related to flavonoid content in tea plants.