Search

Many per- and poly-fluoroalkyl substances (PFAS) studies in environmental media have been conducted in heavily industrialized and densely populated areas. However, there has been limited research on legacy and emerging PFAS contamination in cold and arid regions. In the present study, we investigated the concentrations of 30 PFAS compounds in the surface water and sediment of Hulun Lake and its inflowing rivers. The main components in water were short chain perfluoroalkyl carboxylic acids (PFCAs) in both Hulun Lake (33.57 %) and its inflowing rivers (30.47 %). However, long chain perfluoroalkane sulfonates (PFSAs) and long chain PFCAs accounted for more than half of the total PFAS content in sediment. Total PFAS concentrations (∑30PFAS) ranged from 3.67 to 8.84 ng/L in water, and 0.97–1.73 μg/kg in sediment. Significant spatial differences were apparent between Hulun Lake and its inflowing rivers in both water and sediment samples. Source apportionment revealed that wastewater, aqueous film forming foams, textiles and paper-based food packaging, and paper products and cosmetics were the primary sources of PFAS. The partitioning coefficients of PFCAs were dependent on the carbon chain length. Temperature, conductivity, pH, salinity, chemical oxygen demand, total phosphorus (TP), total nitrogen (TN), As, and oxidation-reduction potential were the main influencing factors in water. Heavy metals (Co, Pb, Cu, Ni, Hg, Cd, and As), TP, salinity, and pH were positively correlated with the PFAS concentration in sediment. Furthermore, Hailar River contributed the majority (99.74 %, ∼15.05 kg/year) of the PFAS mass flux. This is the first study showing PFAS contamination in Hulun Lake and the results suggest that long term monitoring is needed for the effective control of PFAS pollution in this typical cold and arid region of China.

ABSTRACTIn multi-robot cooperative wire and arc additive manufacturing (MRC-WAAM) systems, allocating and scheduling deposition tasks is necessary. The challenge comes from the factorial computational complexity when optimising robot time, workload, and deposition order. This work presents a novel method, ‘top k%’. The main idea is to iteratively assign k% amount of paths from a warehouse to the robot having the least workload until all paths are set. A collision-free deposition process is scheduled by checking the overlapping conditions of robot swap areas. Both simulative testing and actual depositions were conducted to validate the applicability and effectiveness of the method. The results indicated that the ‘top k%’ method can facilitate the equality of robot workload and enhance the adjacency of paths. The top k% method outperforms the existing solutions concerning a reduction of turns and start-arc points, which improves the forming quality of MRC-WAAM.

ABSTRACTCurved layering has become one of the current research hotspots for its unique advantages. In order to improve the adaptability and accuracy, this paper proposes a new curved layering and path planning algorithm based on voxelization and geodesic distance. High density discrete points are used to approximate the shortest path, voxelization and new optimized search algorithm are adopted to obtain the geodesic distance field within stereolithography (STL) model. Curved layers are formed from points with equal geodesic distance values. The developed algorithm overcomes the limitations of the existing facet offsetting method. Moreover, robot positioner angle planning strategies are exploited to achieve good formation for support-free curved layers of spatial overhang characteristics. The results of comparative analysis and verification experiment indicate that the algorithm accuracy is not affected by the size and intersection angle of the triangles, satisfies the high-precision requirements for parts with curved surface in robotic GMA additive manufacturing.

Most directed energy deposition (DED) implementations employ a rigid end effector to deposit material layer-by-layer, which is insufficient in cases with space constraints. To enhance the flexibility, this paper proposes a novel soft-robotic gun (SRG) based DED technology. The ideology is to replace the rigid weld gun with a tendon-driven omnidirectional-steering continuum manipulator, which a computer can numerically control. To this end, both requirements and concepts of the SRG-DED are introduced. A prototype system is designed and implemented before dedicated kinematic modelling and trajectory planning mechanisms are presented. As demonstrated by the experimental results, the developed trajectory planning algorithm improves the computational efficiency of tendon variables and enhances the consistency of trajectory tracking. The prototype system can deposit a complex path on an inclined substrate and fabricate a thin-wall component within a pipeline. The validation work confirms the feasibility and applicability of the prototype system for metal additive manufacturing applications.

Sensory evaluation, biochemical analysis, ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS) and headspace solid-phase microextraction combined with gas chromatography-time-of-flight mass spectrometry (HS-SPME-GC-TOF-MS) were used to detect and analyze different baking degrees of ‘Chungui’ Minbei oolong tea. Partial least squares discriminant analysis model and trend model were established to explore the differences in sensory quality, conventional biochemical components, catechin components, amino acid components and volatile substances. The results showed that the total score of sensory evaluation of light baking degree of ‘Chungui’ (91.3~93.65) was slightly lower than that of medium baking degree (93.6~94.15). Among the biochemical components, the amino acid content of light baking degree was significantly higher than that of medium baking degree (P

Not available.

Abstract Background The formation and domestication of ornamental traits are influenced by various aspects, such as the recognition of esthetic values and cultural traditions. Camellia japonica is widely appreciated and domesticated around the world mainly due to its rich variations in ornamental traits. Ornamental camellias have a diverse range of resources, including different bud variations from Camellia spp. as well as inter- and intra- specific hybridization. Despite research on the formation of ornamental traits, a basic understanding of their genetics and genomics is still lacking. Results Here, we report the chromosomal-level reference genome of C. japonica through combining multiple DNA-sequencing technologies and obtain a high-density genetic linkage map of 4255 markers by sequencing 98 interspecific F1 hybrids between C. japonica and C. chekiangoleosa. We identify two whole-genome duplication events in C. japonica: one is a shared ancient γ event, and the other is revealed to be specific to genus Camellia. Based on the micro-collinearity analysis, we find large-scale segmental duplication of chromosome 8, resulting to two copies of the AGAMOUS loci, which may play a key role in the domestication of floral shapes. To explore the regulatory mechanisms of seasonal flowering, we have analyzed year-round gene expression patterns of C. japonica and C. azalea—a sister plant of continuous flowering that has been widely used for cross breeding. Through comparative analyses of gene co-expression networks and annual gene expression patterns, we show that annual expression rhythms of some important regulators of seasonal growth and development, including GIGANTEA and CONSTANS of the photoperiod pathway, have been disrupted in C. azalea. Furthermore, we reveal that the distinctive expression patterns of FLOWERING LOCUS T can be correlated with the seasonal activities of flowering and flushing. We demonstrate that the regulatory module involved in GIGANTEA, CONSTANS, and FLOWERING LOCUS T is central to achieve seasonality. Conclusions Through the genomic and comparative genomics characterizations of ornamental Camellia spp., we propose that duplication of chromosomal segments as well as the establishment of gene expression patterns has played a key role in the formation of ornamental traits (e.g., flower shape, flowering time). This work provides a valuable genomic platform for understanding the molecular basis of ornamental traits.

Incremental randomized neural networks have been widely applied in industrial data modeling. However, incremental randomized neural networks may generate redundant hidden nodes. These nodes may lead to weak performance in real world data modeling tasks. An important factor is that the neural nodes constructed in the early stages may influence the quality of subsequently generated nodes. To resolve this drawback, an incremental regularization kernel randomized neural network (IRKRNN) is proposed in this work. IRKRNN adopts a kernel learning method to enhance the feature expression during the parameter-learning. Meanwhile, the uncertainty of random mapping is reduced. Each randomly generated node is projected into kernel space. When new nodes are generated, the kernel space is constantly updated. Moreover, the number of nodes and the expected tolerance are used as criteria for terminating the kernel-based incremental learning process to achieve a compact network structure. Finally, IRKRNN is compared with state-of-the-art randomized neural networks on six real world regression datasets and the electrical energy output prediction task of the combined cycle power plant. Experimental results indicate that the proposed IRKRNN achieves satisfactory generalization performance, and it has good potential for industrial data modeling tasks.

Chest X-ray is one of the most widely used methods for clinical diagnosis of chest diseases. In recent years, the development of deep learning technologies has driven progress in chest disease detection, but existing methods still face numerous challenges. Current research primarily focuses on detecting specific chest diseases. However, when chest X-ray images indicate multiple diseases, the diverse and complex characteristics of different disease types make it challenging to extract effective information. Additionally, the detection accuracy of small lesions remains low, which lessens the overall lesion detection rate. To address these issues, a novel network, named YOLO-CXR, is proposed in this paper for multiple disease detection, which is able to effectively locate multiple small lesions in chest X-ray images. Firstly, the proposed network enhances the YOLOv8s backbone by replacing the ordinary convolutional layers with RefConv layers to improve its feature extraction capabilities w.r.t. various diseases. Secondly, it utilizes a novel Efficient Channel and Local Attention (ECLA) mechanism to increase its sensitivity to the spatial location information of different lesions. Thirdly, to enhance its detection of small lesions, YOLO-CXR incorporates a dedicated small-lesion detection head and the Selective Feature Fusion (SFF) technique. Due to these improvements, the proposed network significantly enhances its detection of lesions at different scales and multiple small lesions in particular. Experiments conducted on the publicly available VinDr-CXR dataset demonstrate that YOLO-CXR achieves an mAP@0.5 of 0.338, a mAP@[0.5:0.95:0.05] of 0.167, and recall of 0.365, outperforming all state-of-the-art networks considered.

‘Ming guan’ is a new excellent albino tea variety bred from the descendants of Bai jiguan. In order to explore the quality difference of Ming guan multi tea processing, the fresh leaves of Ming guan were used as raw materials to make the corresponding tea types according to the processing methods of green tea, black tea and white tea, and sensory quality evaluation, aroma and taste analysis were conducted. The results showed that the aroma of Ming guan green tea was tender, floral and fruity, with a mellow taste, the aroma of Ming guan black tea was sweet, floral, with a sweet taste, the aroma of Ming guan white tea was millets, floral, with a fresh taste, and the different processes of Ming guan tea had their own unique floral characteristics. Among the aroma components of Ming guan green tea, terpene aroma components with floral aroma were relatively more abundant, followed by ester aroma components with fruit aroma, which played an important role in the formation of the aroma of Ming guan green tea. The representative aroma components of Ming guan green tea were leaf alcohol ester of foliol caproate, 3-hexenyl caproate, 2-hexenyl caproate, nerolidol, leaf alcohol ester of butyric acid, olivetol and α-farnesene, the representative components of which were mainly esters with fruity aroma and alcohols with floral and fruity aroma, creating the characteristic of Ming guan green tea floral and fruity varieties. The representative components of Ming guan black tea were dihydrolinalool, α-cephalene, β-Ionone, γ-cadinene, methyl hexadecanoic acid and benzaldehyde, which were mainly terpenes and alcohols with floral and sweet aromas, contributing to the floral and sweet aromatic characteristics of Ming guan black tea. The representative components of Ming guan white tea were geraniol, myrcene, 3-carene, linalyl acetate and linalool, and the representative components are mainly alcohols and terpenes with floral aroma. The non-volatile components of Ming guan green tea, Ming guan black tea and Ming guan white tea vary greatly overall. The content of catechins, anthocyanins, some flavonols and flavonoid glycosides (quercetin-3-O-galactoside, quercetin-3-O-glucoside, quercetin-3-O-glucoside 7-O-rhamnoside, etc.) in Ming guan green tea was generally higher than that in Ming guan black tea and Ming guan white tea. The contents of theaflavins, phenolic acids, a few flavonol and flavonoid glycoside compounds (vitexin-2-O-galactoside, vitexin-2-O-rhamnoside, apigenin-6,8-di-C-glucoside, apigenin-6-C-glucoside, etc.) and some amino acid compounds (L-phenylalanine, L-tryptophan, L-isoleucine, L-valine, L-aspartic acid) in Ming guan black tea were higher than those in Ming guan green tea and Ming guan white tea.The content of some amino acid compounds (L-arginine, L-glutamine, L-lysine, L-histidine, L-tyrosine) in Ming guan white tea was higher than that in Ming guan green tea and Ming guan black tea, which may be affected by different processing technologies. This study could provide a theoretical basis for a comprehensive understanding of the chemical basis and quality differences of Ming guan green tea, Ming guan black tea and Ming guan white tea.

The complex phylogenetic relationship of polyploid species provides an opportunity for a comprehensive study of gene introgression. Oil camellias refer to a class of important woody oil plant in the camellia genus, including octoploid, hexaploid, tetraploid, and diploid plants, but the phylogeny relationship of these species remains poorly investigated. Here, based on multiple types of evidence, including phylogenetic conflict, gene flow analysis, and representative metabolite, we reconstructed the phylogenetic relationship of oil camellias. Camellia shensiensis and C. grijsii formed a distinct branch. Phylogenetic conflict suggested that hexaploid C. oleifera probably originated from hybridization and clustered with diploid C. kissi and tetraploid C. meiocarpa. Tetraploid C. confusa probably originated from crossing the ancestor of C. kissi and C. brevistyla, and C. brevistyla probably was the maternal progenitor of hexaploid C. sasanqua. Furthermore, the composition of anthocyanin in tender leaves showed a strong correlation with phylogenetic distinctions. This study proves the feasibility of using iconic metabolic components to solve phylogenetic relationships and lays a foundation for analyzing genetic breeding and utilizing oil camellia resources.

‘Ming guan’ is a new chlorotic tea cultivar bred from the descendants of Bai jiguan, in order to explore the characteristic quality of aroma and taste of Ming guan green tea, this study conducted sensory quality evaluation and metabomic analysis on green tea made from Ming guan and Fuding dabai, to find the characteristic metabolic components of Ming guan green tea. The results showed that the aroma of Ming guan green tea was tender, with a long-lasting floral aroma and a mellow taste; the aroma of Fuding dabai green tea was clear and the taste was mellow. Hexanoate was the aroma group with the highest relative content in Ming guan green tea, accounted for 6.57% of the total content of aroma components. Other volatile components such as geraniol, (Z)-3-hexenyl hexanoate, hotrienol, nerolidol, (Z)-2-hexenyl hexanoate, α-cedrene, α-farnesene, hexenyl butyrate and laurene were significantly higher than those of Fuding dabai green tea (P

Abstract Dexterous locomotion, such as immediate direction change during fast movement or shape reconfiguration to perform diverse tasks, are essential animal survival strategies which have not been achieved in existing soft robots. Here, we present a kind of small-scale dexterous soft robot, consisting of an active dielectric elastomer artificial muscle and reconfigurable chiral-lattice foot, that enables immediate and reversible forward, backward and circular direction changes during fast movement under single voltage input. Our electric-driven soft robot with the structural design can be combined with smart materials to realize multimodal functions via shape reconfigurations under the external stimulus. We experimentally demonstrate that our dexterous soft robots can reach arbitrary points in a plane, form complex trajectories, or lower the height to pass through a narrow tunnel. The proposed structural design and shape reconfigurability may pave the way for next-generation autonomous soft robots with dexterous locomotion.

Abstract Cancers are a group of heterogeneous diseases characterized by the acquisition of functional capabilities during the transition from a normal to a neoplastic state. Powerful experimental and computational tools can be applied to elucidate the mechanisms of occurrence, progression, metastasis, and drug resistance; however, challenges remain. Bulk RNA sequencing techniques only reflect the average gene expression in a sample, making it difficult to understand tumor heterogeneity and the tumor microenvironment. The emergence and development of single-cell RNA sequencing (scRNA-seq) technologies have provided opportunities to understand subtle changes in tumor biology by identifying distinct cell subpopulations, dissecting the tumor microenvironment, and characterizing cellular genomic mutations. Recently, scRNA-seq technology has been increasingly used in cancer studies to explore tumor heterogeneity and the tumor microenvironment, which has increased the understanding of tumorigenesis and evolution. This review summarizes the basic processes and development of scRNA-seq technologies and their increasing applications in cancer research and clinical practice.

Abstract. Background:. Exploring the underlying mechanism of rituximab resistance is critical to improve the outcomes of patients with diffuse large B-cell lymphoma (DLBCL). Here, we tried to identify the effects of the axon guidance factor semaphorin-3F (SEMA3F) on rituximab resistance as well as its therapeutic value in DLBCL. Methods:. The effects of SEMA3F on the treatment response to rituximab were investigated by gain- or loss-of-function experiments. The role of the Hippo pathway in SEMA3F-mediated activity was explored. A xenograft mouse model generated by SEMA3F knockdown in cells was used to evaluate rituximab sensitivity and combined therapeutic effects. The prognostic value of SEMA3F and TAZ (WW domain-containing transcription regulator protein 1) was examined in the Gene Expression Omnibus (GEO) database and human DLBCL specimens. Results:. We found that loss of SEMA3F was related to a poor prognosis in patients who received rituximab-based immunochemotherapy instead of chemotherapy regimen. Knockdown of SEMA3F significantly repressed the expression of CD20 and reduced the proapoptotic activity and complement-dependent cytotoxicity (CDC) activity induced by rituximab. We further demonstrated that the Hippo pathway was involved in the SEMA3F-mediated regulation of CD20. Knockdown of SEMA3F expression induced the nuclear accumulation of TAZ and inhibited CD20 transcriptional levels via direct binding of the transcription factor TEAD2 and the CD20 promoter. Moreover, in patients with DLBCL, SEMA3F expression was negatively correlated with TAZ, and patients with SEMA3F lowTAZ high had a limited benefit from a rituximab-based strategy. Specifically, treatment of DLBCL cells with rituximab and a YAP/TAZ inhibitor showed promising therapeutic effects in vitro and in vivo. Conclusion:. Our study thus defined a previously unknown mechanism of SEMA3F-mediated rituximab resistance through TAZ activation in DLBCL and identified potential therapeutic targets in patients.

In order to explore the processing technology characteristics of northern Fujian oolong tea processed by Chungui variety, sensory evaluation, biochemical analysis, ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS) and headspace solid-phase microextraction combined with gas chromatography-time of flight mass spectrometry (HS-SPME-GC-TOF-MS) were used to investigate the differences in sensory quality, physicochemical quality, catechin fraction, amino acid fraction and volatile substances of northern Fujian oolong tea at different turning over intensities. The results showed that the light turning over intensity treatment had tight shape, with a distinctive floral aroma and fresh taste, and the total score of sensory quality was 90.6, and heavy turning over intensity treatment had slightly loose shape, floral fragrance and a strong mellow taste, sensory score 88.0. The catechin components were all significantly higher in the light turning over intensity treatment of Chungui than heavy turning over intensity treatment (P

Camellia nitidissima is famous for its golden flowers. Its flowers are rich in secondary metabolites, and they have ornamental, medicinal, and edible value. Pigment composition and regulation has been studied in the golden petals, but there has been little research on pigment composition or the molecular mechanism underlying yellow stamens in C. nitidissima. To explore the molecular mechanism of yellow stamen formation, three developmental stages (S0, S1, and S2) were used for transcriptome and pigment analyses. Pigment analysis showed that the flavonoid content increased sharply from the S0 to S1 stage and decreased from the S1 to S2 stage, and the carotenoid content increased sharply during yellow stamen formation (from the S1 to S2 stage). RNA-seq analysis showed that a total of 20,483 differentially expressed genes (DEGs) were identified. KEGG and heatmap analyses showed that flavonoid and carotenoid biosynthesis pathways were enriched, and we identified 14 structural genes involved in flavonoid biosynthesis and 13 genes involved in carotenoid biosynthesis and degradation. In addition, the expression of carotenoid- and flavonoid-related genes was consistent with carotenoid and flavonoid content. In addition, correlation network analysis indicated that the WARYK, MYB, bHLH, and AP2/ERF transcription factor families were screened for involvement in the biosynthesis of flavonoids and carotenoids. In this study, we describe the pathway associated with color formation in the stamens of C. nitidissima.

FLOWERING LOCUS T (FT), belonging to the FT/TFL1 gene family, is an important gene regulating the flowering transition and inflorescence architecture during plant development. Given its importance to plant adaptation and crop improvement, FT has been extensively studied in related plant research; however, the specific role and underlying molecular mechanisms of FT in the continuous flowering of perennial plants remains elusive. Here, we isolated and characterized homologous FT genes from two Camellia species with different flowering-period phenotypes: CaFT was isolated from Camellia azalea, a precious species blooming in summer and flowering throughout the year, and CjFT was isolated from C. japonica, which blooms in winter and spring. The major difference in the genes between the two species was an additional five-amino acid repeat sequence in C. japonica. FT showed high expression levels in the leaves in both species from January to August, especially in April for C. japonica and in May for C. azalea. CaFT was expressed throughout the year in C. azalea, whereas CjFT was not expressed from September to December in C. japonica. The expression levels of FT in the floral buds were generally higher than those in the leaves. Overexpression of CaFT and CjFT in Arabidopsis indicated that both genes can activate downstream genes to promote flowering. Transgenic callus tissue was obtained by introducing the two genes into C. azalea through Agrobacterium-mediated transformation. Transcriptome and quantitative real-time polymerase chain reaction analyses indicated that both florigen FT genes promoted the expression of downstream genes such as AP1, FUL, and SEP3, and slightly up-regulated the expression of upstream genes such as CO and GI. The above results indicated that CaFT and CjFT played a role in promoting flowering in both camellia species. The expression pattern of CaFT in leaves suggested that, compared to CjFT, CaFT may be related to the annual flowering of C. azalea.

Abstract Animal‐derived anti‐IgG secondary antibodies are currently employed to stain and screen of human monoclonal antibody(mAb)‐producing cells, but using animal‐derived antibodies may raise the concerns of high cost, complicated operations and biological safety issues in biopharmaceutical manufacturing. Nanobodies(VHHs) are attractive forms of antibodies for their straightforward engineering and expression in both eukaryotic and prokaryotic systems. Using phage‐displayed immune llama VHH library, we identified new anti‐Fc VHHs that could bind to human Fc with high affinity. In GFP fusion format, the anti‐Fc VHH‐GFP generated dramatically stronger FACS signals than AF488 conjugated anti‐IgG antibodies when used for staining mAb‐producing CHO cells. Furthermore, preparative sorting of CHO cells based on anti‐Fc VHH‐GFP staining resulted in the enrichment of cell lines capable of synthesizing mAb at high productivity. This safe and cost‐efficient anti‐Fc VHH‐GFP may optimize the process of generating highly productive cell lines for therapeutic mAb production compared to conventional animal‐derived fluorescent antibodies.

Abstract Background The cultivated Camellia sasanqua forms a divergent double flower pattern, and the stamen petaloid is a vital factor in the phenomenon. However, the regulation mechanism remains largely unclear. Results Here, a comprehensive comparative transcriptome analysis of the wild-type, “semi-double”, “peony double”, and “rose double” was performed. The cluster analysis of global gene expression level showed petal and stamen difficulty separable in double flower. The crucial pathway and genes related to double flower patterns regulation were identified by pairwise comparisons and weighted gene coexpression network (WGCNA). Divergent genes expression, such as AUX1 and AHP, are involved in plant hormone signaling and photosynthesis, and secondary metabolites play an important role. Notably, the diversity of a petal-specific model exhibits a similar molecular signature to the stamen, containing extensin protein and PSBO1, supporting the stamen petaloid point. Moreover, the expansion of class A gene activity influenced the double flower formation, showing that the key function of gene expression was probably demolished. Conclusions Overall, this work confirmed the ABCE model and provided new insights for elucidating the molecular signature of double formation.

The use of ‘Electrostatic tweezers’ is a promising tool for droplet manipulation, but it faces many limitations in manipulating droplets on superhydrophobic surfaces. Here, we achieve noncontact and multifunctional droplet manipulation on Nepenthes-inspired lubricated slippery surfaces via triboelectric electrostatic tweezers (TETs). The TET manipulation of droplets on a slippery surface has many advantages over electrostatic droplet manipulation on a superhydrophobic surface. The electrostatic field induces the redistribution of the charges inside the neutral droplet, which causes the triboelectric charged rod to drive the droplet to move forward under the electrostatic force. Positively or negatively charged droplets can also be driven by TET based on electrostatic attraction and repulsion. TET enables us to manipulate droplets under diverse conditions, including anti-gravity climb, suspended droplets, corrosive liquids, low-surface-tension liquids (e.g. ethanol with a surface tension of 22.3 mN·m ^−1 ), different droplet volumes (from 100 nl to 0.5 ml), passing through narrow slits, sliding over damaged areas, on various solid substrates, and even droplets in an enclosed system. Various droplet-related applications, such as motion guidance, motion switching, droplet-based microreactions, surface cleaning, surface defogging, liquid sorting, and cell labeling, can be easily achieved with TETs.

Abstract Background As a perennial crop, oil-Camellia possesses a long domestication history and produces high-quality seed oil that is beneficial to human health. Camellia oleifera Abel. is a sister species to the tea plant, which is extensively cultivated for edible oil production. However, the molecular mechanism of the domestication of oil-Camellia is still limited due to the lack of sufficient genomic information. Results To elucidate the genetic and genomic basis of evolution and domestication, here we report a chromosome-scale reference genome of wild oil-Camellia (2.95 Gb), together with transcriptome sequencing data of 221 cultivars. The oil-Camellia genome, assembled by an integrative approach of multiple sequencing technologies, consists of a large proportion of repetitive elements (76.1%) and high heterozygosity (2.52%). We construct a genetic map of high-density corrected markers by sequencing the controlled-pollination hybrids. Genome-wide association studies reveal a subset of artificially selected genes that are involved in the oil biosynthesis and phytohormone pathways. Particularly, we identify the elite alleles of genes encoding sugar-dependent triacylglycerol lipase 1, β-ketoacyl-acyl carrier protein synthase III, and stearoyl-acyl carrier protein desaturases; these alleles play important roles in enhancing the yield and quality of seed oil during oil-Camellia domestication. Conclusions We generate a chromosome-scale reference genome for oil-Camellia plants and demonstrate that the artificial selection of elite alleles of genes involved in oil biosynthesis contributes to oil-Camellia domestication.

Extranodal NK/T-cell lymphoma (ENKTL) is a highly aggressive and heterogeneous disease with poor clinical outcome. Our previous work had demonstrated that circulating tumor DNA (ctDNA) analyses were feasible in ENKTL, and dynamic tracing of ctDNA could be used to monitor the disease status. However, the prognostic value of ctDNA in ENKTL has not been fully investigated. Patients with newly diagnosed ENKTL from February 2017 to December 2021 (n = 70) were enrolled. The pretreatment ctDNA concentration (hGE/mL) was measured. The prognostic value of ctDNA, international prognostic index (IPI), Korean prognostic index (KPI), PINK-E, and the combination of PINK-E and ctDNA (PINK-EC) were investigated in our cohort. The IPI and PINK-E risk categories had a significant difference in progression-free survival (PFS) and overall survival (OS) between the low-risk and intermediate-risk groups. The KPI risk category had a difference in PFS and OS between the intermediate-risk and high-risk groups. Furthermore, integrating ctDNA into the PINK-E model could overcome the shortcomings of other prognostic models, which could significantly distinguish the different-risk groups. Overall, our results demonstrated that PINK-EC showed a superior prognostic prediction value and stability compared with IPI, KPI, and PINK-E. The integration of molecular features of the tumor into classic risk categories might better characterize a high-risk group where novel treatment approaches are most needed.

The effect of Mn content on the microstructure evolution and mechanical properties of Al–Cu–Mg–x Mn alloys at ambient temperature was investigated. The findings show that in the Mn-containing alloys at the as-cast state, the blocky primary T(Al20Cu2Mn3) phase coexisting with the Al2Cu phase appeared. With the increase in Mn content, the majority of the Al2Cu phase dissolved, nd a minor amount of the T phase remained at the grain boundary after solution treatment. The rod-like TMn (Al20Cu2Mn3) nanoprecipitate was simultaneously distributed at grain boundaries and the interiors, while a high density of needle-like θ″ (Al3Cu) nanoprecipitate was also observed in the T6 state. Further increases in Mn content promoted the dispersion of the TMn phase and inhibited the growth and transformation of the θ″ phase. Tensile test results show that 0.7 wt.% Mn alloy had excellent mechanical properties at ambient temperature with ultimate tensile strength, yield strength, and fracture elongation of 498.7 MPa, 346.2 MPa, and 19.2%, respectively. The subsequent calculation of strengthening mechanisms elucidates that precipitation strengthening is the main reason for the increase in yield strength of Mn-containing alloys.

The strength-ductility trade-off severely limits the extensive application of particle-reinforced aluminum matrix composites mainly owing to the critical issues of local particle agglomeration and poor interfacial characteristics. The present work concentrates on these intractable puzzles and proposes Mg-Ag microalloying TiB2/Al-4.5Cu composites fabricated by an optimized salt-metal reaction method and multistage heat treatment. The resulting microstructure evidence that this new tactic synergistically ameliorates the detrimental agglomeration of TiB2 particles by Mg and Ag doped at the TiB2/α-Al interface， meanwhile promoting the formation of nanosized Ω-Al2Cu and σ-Al5Cu6Mg2 phases. The mechanical property experimental results indicate that the Mg-Ag microalloyed TiB2/Al-4.5Cu composites exhibit an accredited combination of the tensile strength (ultimate tensile strength is 496.3 ± 15.0 MPa) and ductility (uniform elongation is 12.8 ± 1.4 %). We emphasize the reciprocal effect between TiB2 particles and the precipitated phase associated with unrelaxed misfit strains. Close attention is also paid to the interface binding characteristics and semi-coherent interphase boundary to evaluate their contribution to the ductility. The microalloying strategy can potentially be applied to many other metal matrix composites and boost analogous approaches for designing ultrahigh yet ductile materials for technological applications.

Given the shortcomings of conventional machine vision-based surface defect detection methods, including their low accuracy, long development cycle, and poor generalization ability, this paper proposes a surface defect detection model based on the convolutional block attention module and atrous convolution. This model combines the surface defect segmentation task of the product with the classification task, obtains contextual information of the image at multiple scales using atrous spatial pyramid pooling, and then uses the convolutional block attention module to reallocate the weighting of the network to enhance focus on the defect area and improve the discrimination of extracted features. In addition, atrous convolution was introduced in the deep network to simplify the model when used in defect segmentation tasks and enhances the real-time performance of the model defect detection method. Experiments show the superior accuracy and real-time performance of the proposed model when compared with current mainstream surface defect detection methods and indicate its wide applicability in the detection of surface defects in industrial products.

Abstract Background The sodium taurocholate cotransporting polypeptide (NTCP) is a functional receptor for hepatitis B virus (HBV). NTCP-reconstituted human hepatoma cells support HBV infection, but the infection is suboptimal and no apparent HBV spread has been observed in this system. Results We found that NTCP-reconstituted HepG2 cells were highly susceptible to HBV infection after cells were cultured in a commercial human inducible pluripotent stem cell (iPSC)-derived hepatocyte maintenance medium (HMM). The enhanced HBV infection coincided with increased NTCP expression, and was observed in six different clones of HepG2-NTCP cells. Promoter assays indicated that HMM activated the cytomegalovirus immediate-early (IE) promoter that drives the NTCP expression in the HepG2-NTCP cells. RNA-Seq analysis revealed that HMM upregulated multiple metabolic pathways. Despite highly upregulated NTCP expression by HMM, no obvious HBV spread was observed even in the presence of PEG 8000. Conclusions Our data suggest that this particular medium could be used to enhance HBV infection in NTCP-reconstituted hepatocytes in vitro.

High-temperature thermal stimulation (HTS) can effectively remove the damage caused by water phase trapping during the drilling and completion of tight oil and hydraulic fracturing of gas reservoirs. Since HTS induces new rock fracture, the permeability of a reservoir will be increased to a certain extent. However, there are no standardized methodologies for pretreatment, experimental procedures, or evaluation indicators for HTS rock samples. Taking the Longmaxi Formation shale from the Sichuan Basin as the research object, an experimental process for evaluating the impact of HTS on permeability was established. The temperature increased at a rate of 5 °C/min, considering the conditions of dry and water-bearing rock samples. Three variables are included in the evaluation index for HTS of shale reservoirs: threshold temperature, raise multiples of permeability in 4 MPa confining pressure, and in-situ conditions. The results show that the threshold temperature of the dry shale is 650–700 °C, while the water-bearing shale has two threshold temperatures. Furthermore, the low temperature is 100–150 °C, while the high temperature is 450–500 °C. The permeability increase ratio of drying shale samples is 1.5–10.0 after HTS under in-situ effective stress, while that of water-bearing shale samples reach 20–50. This research has shown that using the HTS method to reduce water phase trapping damage and drastically improves well production are being conducted in a shale gas reservoir with horizontal wells and staged hydraulic fracturing. Our recommended experimental methods and evaluation indicators are effective in establishing the HTS potential of a shale reservoir.

Several methods have been used to assess heavy metal contamination in sediments. However, an assessment that considers both composite heavy metal speciation and concentration is necessary to accurately study ecological risks. This study improved the potential ecological risk index method and calculated the toxicity coefficients of seven heavy metals: Arsenic (As), Cadmium (Cd), Chromium (Cr), Copper (Cu), Nickel (Ni), Lead (Pb), and Zinc (Zn). The newly calculated toxicity coefficients were validated by using previously published heavy metal distribution data of the Henan section of the Yellow River. The calculation procedure is based on the principle that the abundance of heavy metals in the environment and their bioavailable forms affect the toxicity of heavy metals. The toxicity coefficients for the seven heavy metals were calculated as follows: As = 10, Cd = 20, Cr = 5, Cu = 2, Ni = 5, Pb = 5, Zn = 1. Ecological risk assessment of the Henan section of the Yellow River using the improved toxicity coefficients revealed that the ecological risk of Cd and total heavy metals is higher than previous calculations, reaching the strength and moderate risk levels, respectively. The improved potential ecological risk index method is more sensitive to heavy metal pollution and thus provides a better indication of ecological risk. This is a necessary improvement to provide more accurate pollution assessments.

Flexible mechanical sensors based on nanomaterials operate on a deformation-response mechanism, making it challenging to discern different types of mechanical stimuli such as pressure and strain. Therefore, these sensors are susceptible to significant mechanical interference. Here, we introduce a multifunctional flexible sensor capable of discriminating coupled pressure and strain without cross-interference. Our design involves an elastic cantilever fixed on the pillar of the flexible main substrate, creating a three-dimensional (3D) substrate, and two percolative nanoparticle (NP) arrays are deposited on the cantilever and main substrate, respectively, as the sensing materials. The 3D flexible substrate could confine pressure/strain loading exclusively on the cantilever or main substrate, resulting in independent responses of the two nanoparticle arrays with no cross-interference. Benefitting from the quantum transport in nanoparticle arrays, our sensors demonstrate an exceptional sensitivity, enabling discrimination of subtle strains down to 1.34 × 10−4. Furthermore, the suspended cantilever with one movable end can enhance the pressure perception of the NP array, exhibiting a high sensitivity of −0.223 kPa−1 and an ultrahigh resolution of 4.24 Pa. This flexible sensor with multifunctional design will provide inspiration for the development of flexible mechanical sensors and the advancement of decoupling strategies.

Previously, we showed that mouse immunity-related guanosine triphosphatase (GTPase) family M protein 1 (Irgm1) promotes malignant melanoma progression by inducing cellular autophagy flux and metastasis. Human IRGM, a truncated protein functionally distinct from its mouse counterpart, has several splice isoforms. In this study, we analyzed the association of IRGM and human melanoma clinical prognosis and investigated the function of IRGM in human melanoma cells. Data from the training cohort (n = 144) showed that overexpression of IRGM is proportional to melanoma genesis and clinical stages in human tissue chips. A validation cohort (n = 78) further confirmed that IRGM is an independent risk factor promoting melanoma progression and is associated with poor survival of patients. Among IRGM isoforms, we found that IRGMb is responsible for such correlation. In addition, IRGM promoted melanoma cell survival through autophagy, both in vitro and in vivo. We further showed that the blockade of translocation of high-mobility group box 1 (HMGB1) from the nucleus to cytoplasm inhibits IRGM1-mediated cellular autophagy and reduces cell survival. IRGM functions as a positive regulator of melanoma progression through autophagy and may serve as a promising prognostic marker and therapeutic target.

Sevoflurane anesthesia induces cognitive impairment, which may lead to perioperative neurocognitive disorders (PND). However, the factors and molecular mechanism underlying this impairment remains unclear. Adult hippocampal neurogenesis (AHN) in the subgranular zone of the hippocampus has been implicated in cognitive processes. Nonetheless, the direct role of AHN in sevoflurane-induced cognitive impairment has never been demonstrated. In this study, we explored the age and the concentration factors and the role of AHN inhibition in sevoflurane-induced cognitive impairment in sevoflurane inhalation model mice. We found that 3% sevoflurane exposure induced significant cognitive impairment and inhibition of AHN in aged mice but not adult mice. Expression of BDNF/TrkB and NT-3/TrkC was also decreased by 3% sevoflurane exposure in aged mice. Hippocampal brain-derived neurotrophic factor (BDNF) or Neurotrophin-3 (NT-3) microinjection could partially improve the sevoflurane-induced cognitive impairment and AHN inhibition, respectively. These results demonstrate that the cognitive impairment caused by sevoflurane inhalation is related to patient age and sevoflurane concentration. In conclusion, the molecular mechanism of cognitive impairment in the elderly is related to the inhibition of AHN through the BDNF/TrkB and NT-3/TrkC pathways. Thus, sevoflurane inhalation anesthesia may be safe for adult patients, but caution should be exercised when administering it to the elderly.

Understanding the molecular mechanism of the cold response is critical to improve horticultural plant cold tolerance. Here, we documented the physiological, transcriptome, proteome, and hormonal dynamics to cold stress in temperate genotype (Tg) and subtropical genotype (Sg) populations of Camellia japonica. Tg C. japonica suffered minimal osmotic and oxidative damage compared to Sg C. japonica under the same cold treatment. Transcriptional and translational differences increased under the cold treatment, indicating that Tg C. japonica was affected by the environment and displayed both conserved and divergent mechanisms. About 60% of the genes responding to cold had similar dynamics in the two populations, but 1,896 transcripts and 455 proteins differentially accumulated in response to the cold between Tg and Sg C. japonica. Co-expression analysis showed that the ribosomal protein and genes related to photosynthesis were upregulated in Tg C. japonica, and tryptophan, phenylpropanoid, and flavonoid metabolism were regulated differently between the two populations under cold stress. The divergence of these genes reflected a difference in cold responsiveness. In addition, the decrease in the abscisic acid (ABA)/gibberellic acid (GA) ratio regulated by biosynthetic signal transduction pathway enhanced cold resistance in Tg C. japonica, suggesting that hormones may regulate the difference in cold responsiveness. These results provide a new understanding of the molecular mechanism of cold stress and will improve cold tolerance in horticultural plants.

Camellia nitidissima is a woody plant with high ornamental value, and its golden-yellow flowers are rich in a variety of bioactive substances, especially flavonoids, that are beneficial to human health. Chalcone isomerases (CHIs) are key enzymes in the flavonoid biosynthesis pathway; however, there is a scarcity of information regarding the CHI family genes of C. nitidissima. In this study, seven CHI genes of C. nitidissima were identified and divided into three subfamilies by phylogenetic analysis. The results of multiple sequence alignment revealed that, unlike CnCHI1/5/6/7, CnCHI2/3/4 are bona fide CHIs that contain all the active site and critical catalytic residues. Analysis of the expression patterns of CnCHIs and the total flavonoid content of the flowers at different developmental stages revealed that CnCHI4 might play an essential role in the flavonoid biosynthesis pathway of C. nitidissima. CnCHI4 overexpression significantly increased flavonoid production in Nicotiana tabacum and C. nitidissima. The results of the dual-luciferase reporter assay and yeast one-hybrid system revealed that CnMYB7 was the key transcription factor that governed the transcription of CnCHI4. The study provides a comprehensive understanding of the CHI family genes of C. nitidissima and performed a preliminary analysis of their functions and regulatory mechanisms.

In the context of social transformation, the needs of society and individuals for information technology are changing, and discussions on the value and function of information technology are getting more and more attention. The purpose of this research is to clarify the value problems of the information technology curriculum and its function for nurturing students in primary and secondary schools. This research adopts content analysis and comparative research methods, which focus on the process, content selection, and implementation status of information and communication technology (ICT) in the Computing curriculum of British primary and secondary schools. This research content is based on an analysis perspective of curriculum value theory from levels of social and human development, and finally formed the analysis framework of value in information technology curriculum standards. The research results analyze reasons for changes in the value orientation and practical performance of the information technology curriculum standards in Britain to help understand trends and challenges of information technology reform in different contexts. Finally, this research finds the effects of content and value changes of curriculum standards on social and student development and puts forward conclusions for the position and value of information technology in the future of learning and teaching in the context of educational informatization.

Computational thinking (CT) is valued as a thinking process that is required to adapt to the development of curriculum in primary education. In the context of modern information technology, English as a language subject emphasizes the necessity for changes in both learning and teaching modes. However, there is a lack of up-to-date synthesis research and a comprehensive overview surrounding CT integrated into English language curriculum learning and teaching in primary education. To address this research gap, this study conducted a systematic literature review on CT in the primary English curriculum, based on papers published from 2011 to 2021. The purpose of this review is to systematically examine and present empirical evidence on how CT can be integrated into the teaching and learning of the primary English language curriculum in educational contexts. The review was conducted based on the PRISMA 2020 statement and presents a synthesis of 32 articles. The CT-TPACK model was adopted as a lens and framework to analyze these articles. The results indicate that the relationship among CT, content knowledge of English language curriculum, pedagogy and learning knowledge, technology and resources is highlighted. Research on the integration of CT into English courses using unplugged activities is still insufficient. The research about how teachers and students use CT to support content knowledge of the English language curriculum in various educational contexts is still in its infancy.

In the context of digital technologies, computational thinking (CT) is considered a key competence of primary school teachers. However, assessment tools to measure in-service teachers' CT competence are rare. The current study explains the design, development, and validation of a scale to determine the in-service teachers' mastery level of their Computational Thinking Competence (CTC) in the Chinese context. The validity and reliability of the scale have been studied by conducting exploratory factor analysis (EFA), confirmatory factor analysis (CFA) and calculating internal consistency coefficients. The sample of this study consisted of 631 in-service teachers from different Chinese primary schools (EFA N = 189; CFA N = 442). The 31-item scale mirrors five factors: cognitive knowledge of CT, practices and skills of CT, attitudes and perspectives of CT, CT teaching design, and CT teaching implementation. The discussion of the results looks at the future use of the CT scale in Chinese educational settings that mirror a specific national education policy, talent demand, and educational culture.

Facial medical analysis, including the inspection of the face and inner facial components, has always been a primary part of the diagnostic method in Traditional Chinese Medicine (TCM). The existing literature merely focus on detecting or segmenting single face organs such as tongue, eyes, or lips. In this paper, we make the first attempt to deal with multiple organs simultaneously and develop an end-to-end hybrid network with context aggregation (named TCMINet) to achieve face parsing for Traditional Chinese Medicine Inspection (TCMI). Additionally, we construct a new dataset named TCMID to overcome the lackness of accurate annotated data. In order to verify the generalization ability of TCMINet, we manually relabel images in two popular face parsing datasets referred to as LFW-PL* and HELEN* for test. The extensive ablation evaluations and experimental comparisons demonstrate that the proposed TCMINet outperforms state-of-the-art methods under various evaluation metrics. It runs at 267ms per face (512×512 image) on Nvidia Titan Xp GPU, being possible to be integrated into engineering solutions.

The small-screen era has created a new life scene for China’s ‘Internet generation’. With the increase in mobile phone penetration among rural children, entertainment has become the biggest goal of rural children using mobile phones. The mobile phone usage of rural children is mostly concentrated on short videos, WeChat and other information apps. The method of direct prohibition adopted by parents and school is not effective in correcting the child’s attitude towards using the phone. The simple living environment of rural children and the left-behind phenomenon leads to the lack of parental supervision. Systematic analysis of rural children’s attitude towards the use of mobile phones and finding factors affecting their use of mobile phones can effectively guide them to obtain knowledge and achieve anti-sinking purpose. Based on the ABC attitude model combined with face-to-face interviews, this paper firstly determines the influencing variables and designs the questionnaire for collecting the data in the rural schools. With fitting of structural equation model, the attitude model affecting rural children’s use of mobile phones is established. The factors influencing rural children’s mobile phone use are analyzed to improve the essential understanding of the problem.

Deletion and mutation of the Smad4 gene are favorable events for the progression of colon cancer, which is related to the negative regulation of vascular endothelial growth factor C (VEGF-C). However, the regulatory mechanism between Smad4 and VEGF-C remains unclear. We reported first that Smad4 can increase the transcription of miR-128-3p, a microRNA targeting VEGF-C mRNA, resulting in a negative correlation between Smad4 and VEGF-C. Moreover, we found that Smad4 combined with Smad3 can positively regulate VEGF-C during colon cancer metastasis through binding to VEGF-C gene promoter. Further, results revealed a mechanism that long noncoding RNA (lncRNA) ASLNC07322 increased specifically in metastatic colon cancer and decreased miR-128-3p as a sponge, leading to a subsequent elevation of VEGF-C. In a word, there are two pathways in the progression of colon cancer, including Smad4/miR-128-3p/VEGF-C and Smad4/VEGF-C pathways in non-metastatic and metastatic colon cancer, respectively. ASLNC07322 crucially controlled this negative and positive regulatory transformation between them. Additionally, ASLNC07322 knockdown combined with Smad4 overexpression could efficiently inhibit lymphatic endothelial cells (LECs) proliferation and tube formation in vitro, as well as tumor growth and lymphangiogenesis in vivo. These data explained the underlying mechanism of Smad4 contribution on VEGF-C expression during metastasis where ASLNC07322 functions vitally as a switch in colon cancer. Keywords: colon cancer, VEGF-C, lncRNA ASLNC07322, miR-128-3p, Smad4

Collectively migrating tumor cells have been recently implicated in enhanced metastasis of epithelial malignancies. In oral squamous cell carcinoma (OSCC), αv integrin is a crucial mediator of multicellular clustering and collective movement in vitro; however, its contribution to metastatic spread remains to be addressed. According to the emerging therapeutic concept, dissociation of tumor clusters into single cells could significantly suppress metastasis-seeding ability of carcinomas. This study aimed to investigate the anti-OSCC potential of novel endostatin-derived polypeptide PEP06 as a cluster-dissociating therapeutic agent in vitro. Firstly, we found marked enrichment of αv integrin in collectively invading multicellular clusters in human OSCCs. Our study revealed that metastatic progression of OSCC was associated with augmented immunostaining of αv integrin in cancerous lesions. Following PEP06 treatment, cell clustering on fibronectin, migration, multicellular aggregation, anchorage-independent survival and colony formation of OSCC were significantly inhibited. Moreover, PEP06 suppressed αv integrin/FAK/Src signaling in OSCC cells. PEP06-induced loss of active Src and E-cadherin from cell–cell contacts contributed to diminished collective migration of OSCC in vitro. Overall, these results suggest that PEP06 polypeptide 30 inhibiting αv integrin/FAK/Src signaling and disrupting E-cadherin-based intercellular junctions possesses anti-metastatic potential in OSCC by acting as a cluster-dissociating therapeutic agent. KEY WORDS: Oral squamous cell carcinoma, Tumor cell clusters, Collective migration, Metastasis, αv integrin/FAK/RC signaling, RGD

Camellia fluviatilis is an important shrub producing edible seed oil, which is widely cultivated in South China. In this study, the complete chloroplast genome was sequenced and analyzed based on the Illumina HiSeq platform. The results showed that the complete chloroplast genome is 157,041 bp with 37.29% GC content, including a large single copy (LSC) region of 86,718 bp, a small single copy (SSC) region of 18,293 bp, and a pair of inverted repeats (IRs) regions of 26,015 bp. There are 128 genes in the chloroplast genome of C. fluviatilis, including 83 protein-coding genes, 8 ribosomal RNAs, and 37 transfer RNAs. Phylogenetic analysis revealed that C. fluviatilis is closely related to C. lanceoleosa, indicating that both belong to the Sect. Paracamellia Sealy.

Emerging or re-emerging viruses are still major threats to public health. Prophylactic vaccines represent the most effective way to prevent virus infection; however, antivirals are more promising for those viruses against which vaccines are not effective enough or contemporarily unavailable. Because of the slow pace of novel antiviral discovery, the high disuse rates, and the substantial cost, repurposing of the well-characterized therapeutics, either approved or under investigation, is becoming an attractive strategy to identify the new directions to treat virus infections. In this review, we described recent progress in identifying broad-spectrum antivirals through drug repurposing. We defined the two major categories of the repurposed antivirals, direct-acting repurposed antivirals (DARA) and host-targeting repurposed antivirals (HTRA). Under each category, we summarized repurposed antivirals with potential broad-spectrum activity against a variety of viruses and discussed the possible mechanisms of action. Finally, we proposed the potential investigative directions of drug repurposing.

Extracellular vesicles (EVs) are membrane-limited nanoparticles that are liberated by cells and contain a complex molecular payload comprising proteins, microRNA, RNAs, and lipids. EVs may be taken up by other cells resulting in their phenotypic or functional reprogramming. In the liver, EVs produced by non-injured hepatocytes are involved in the maintenance of hepatic homeostasis or therapeutic outcomes following injury while EVs produced by damaged hepatocytes may drive or exacerbate liver injury. In this study, we examined the contribution of EV fibronectin (FN1) to the biogenesis, release, uptake, and action of hepatocyte-derived EVs. While FN1 is classically viewed as a component of the extracellular matrix that regulates processes such as cell adhesion, differentiation, and wound healing and can exist in cell-associated or soluble plasma forms, we report that FN1 is also a constituent of hepatocyte EVs that functions in EV uptake by target cells such as hepatocytes and hepatic stellate cells (HSC). FN1 co-purified with EVs when EVs were enriched from conditioned medium of human or mouse hepatocytes and a direct association between FN1 and hepatocyte EVs was established by immunoprecipitation and proteinase protection. FN1 ablation in mouse hepatocytes using CRISPR-Cas9 did not alter EV biogenesis but EV uptake by HSC was significantly reduced for FN1 knockout EVs (EVΔFN1) as compared to EVs from wild type hepatocytes (EVWT). The uptake by hepatocytes or HSC of either EVWT or EVΔFN1 required clathrin- and caveolin-mediated endocytosis, cholesterol, lysosomal acidic lipase activity, and low pH, while macropinocytosis was also involved in EVΔFN1 uptake in HSC. Despite their differences in rate and mechanisms of uptake, EVΔFN1 functioned comparably to EVWT in ameliorating CCl4-induced hepatic fibrosis in mice. In conclusion, FN1 is a constituent of hepatocyte EVs that facilitates EV uptake by target cells but is dispensable for EV-mediated anti-fibrotic activity in vivo.

Mitochondrial dysfunction contributes to excessive reactive oxygen species (ROS) generation, which is a dramatic cause to promote endothelial dysfunction in diabetes. It was previously demonstrated that crocin protected the endothelium based on its diverse medicinal properties, but its effect on the mitochondrion and the potential mechanism are not fully understood. In this study, mitochondrial function was analyzed during the process of excessive ROS generation in high glucose (HG)-cultured human umbilical vein endothelial cells (HUVECs). The role played by KCa3.1 was further investigated by the inhibition and/or gene silence of KCa3.1 in this process. In addition, nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase 2 (NOX2), superoxide dismutase 1 (SOD1), and glutathione peroxidase 1 (GPx1) were also detected in this study. Our data showed that crocin improved mitochondrial dysfunction and maintained normal mitochondrial morphology by enhancing the mitochondrial membrane potential (MMP), mitochondrial mass, and mitochondrial fusion. Furthermore, KCa3.1 was confirmed to be located in the mitochondrion, and the blockade and/or silencing of KCa3.1 improved mitochondrial dysfunction and reduced excessive ROS generation but did not affect NOX2 and/or the SOD1 system. Intriguingly, it was confirmed that KCa3.1 expression was elevated by ROS overproduction in the endothelium under HG and/or diabetes conditions, while crocin significantly suppressed this elevation by promoting GPx1 and subsequently eliminating ROS generation. In addition, crocin enhanced CD31, thrombomodulin (TM), and p-/t-endothelial nitric oxide synthase (eNOS) expressions as well as NO generation and decreased vascular tone. Hence, crocin improved mitochondrial dysfunction through inhibiting ROS-induced KCa3.1 overexpression in the endothelium, which in turn reduced more ROS generation and final endothelial dysfunction in diabetes.

Camellia chrysanthoides H.T.Chang 1979 is an economic species for its high ornamental and medicinal values. In the present work, the complete chloroplast (cp) genome sequence of C. chrysanthoides was assembled and characterized using Illumina paired-end sequencing data. The total cp genome of C. chrysanthoides is 157,439 bp in size, consisting of a small single copy (SSC) and a large single copy (LSC) separated by a pair of inverted repeats (IRs) with 18,265 bp, 88,162 bp, and 25,506 bp, respectively. Further annotation revealed the cp genome encoded 124 genes, including 82 protein-coding genes, 34 tRNA genes, and eight rRNA genes, and the overall GC content of the cp genome is 37.31%. Phylogenetic analysis based on 80 protein-coding genes shows that C. chrysanthoidesis closely related to C. azalea in the genus Camellia.