Transcriptome sequencing-based analysis of primary vein development in Betula pendula 'Dalecarlica'.

Keymessage The study revealed the major biological processes occurred at three developmental stages and identified candidate genes involved in primary vein development of birch plants. Vascular tissues usually mirror the surrounding leaf shape and its development plays a fundamental role in plant performance. However, the information of vascular development in birch trees, especially primary vein development, remains unclear. Therefore, we conducted the anatomical observation on primary veins from leaves at different development stages in Betula pendula 'Dalecarlica'. With the development of primary vein, dynamic changes in mechanical tissue thickness and primary vein diameter were consistent with each other, and the sum of phloem, xylem and cambium thickness was significantly varied. Transcriptome analysis indicated that primary vein development could be divided into three stages, namely Stage I, II and III, which were in aggreement with anatomical observation. Expression of marker genes associated with vascular tissues revealed that pro-vasculature development occurred at Stage I and II, and phloem development occurred at Stage III. GO enrichment analysis of differentially expressed genes (DEGs) showed that shared DEGs at Stage II were mainly engaged in cell division and cell cycle, and shared DEGs at Stage III were mainly engaged in phosphorylation. Decreased cell division and cell cycle as well as activation of lignin biosynthesis might contribute to primary vein development. Combining phenotypic traits, we performed weighted gene co-expression network analysis and identified a cytochrome P450 84A (CYP84A) family gene (BpF5H1). Based on analyses of gene families, expression patterns and yeast-two hybrid assay results, we proposed a potential electron transfer pathway involving BpF5H1 and three cytochrome b5 proteins during primary vein development in B. pendula 'Dalecarlica'. These results could shed some light on which biological processes occurred during primary vein formation and provide some valuable clues for vascular morphogenesis in woody plants.

BpTCP19 targets BpWRKY53 to negatively regulate jasmonic acid- and dark-induced leaf senescence in Betula platyphylla.

TCP (TEOSINTE-LIKE1, CYCLOIDEA, and PROLIFERATING CELL FACTOR1) is a plant-specific transcription factor that has garnered significant attention due to its wide-ranging involvement in the regulation of plant growth or developmental processes. However, the molecular mechanisms through which TCP genes orchestrate leaf senescence have not been extensively elucidated. BpTCP19, a member of the PCF subfamily in Betula platyphylla, and has high homology to AtTCP19. BpTCP19 displayed pronounced downregulation in response to methyl jasmonate (MeJA) and dark treatment. Overexpressing BpTCP19 in Betula platyphylla led to a delay in leaf senescence, resulting in prolonged leaf greenness under both MeJA and dark conditions. Transcriptome analysis revealed that overexpression of BpTCP19 induced alterations in the expression levels of genes linked to cell proliferation, hormone signaling transduction, and leaf senescence, including the early responsive factor BpWRKY53. Furthermore, through Yeast one-hybrid assays and GUS analysis, BpTCP19 was shown to bind to the promoter region of BpWRKY53, suppressing its expression and thereby retarding leaf senescence. This study elucidates the physiological and molecular functions of BpTCP19 as a central transcriptional regulatory module in leaf senescence and provides a potential target gene for delaying leaf senescence by mitigating sensitivity to external aging signals such as Jasmonic acid (JA) and darkness.

Employing antagonistic C-X-C motif chemokine receptor 4 antagonistic peptide functionalized NaGdF4 nanodots for magnetic resonance imaging-guided biotherapy of breast cancer.

C-X-C motif chemokine receptor 4 (CXCR4) is a promising therapeutic target of breast cancer because it is overexpressed on cell surface of all molecular subtypes of breast cancer including triplenegative breast cancer (TNBC). Herein, CXCR4 antagonistic peptide-NaGdF4 nanodot conjugates (termed as anti-CXCR4-NaGdF4 NDs) have been constructed for magnetic resonance imaging (MRI)-guided biotherapy of TNBC through conjugation of the C-X-C Motif Chemokine 12 (CXCL12)-derived cyclic peptide with tryptone coated NaGdF4 nanodots (5 ± 0.5 nm in diameter, termed as Try-NaGdF4 NDs). The as-prepared anti-CXCR4-NaGdF4 NDs exhibits high longitudinal relaxivity (r1) value (21.87 mM-1S-1), reasonable biocompatibility and good tumor accumulation ability. The features of anti-CXCR4-NaGdF4 NDs improve the tumor-MRI sensitivity and facilitate tumor biotherapy after injection in mouse-bearing MDA-MB-231 tumor model in vivo. MRI-guided biotherapy using anti-CXCR4-NaGdF4 NDs enables to suppress 46% tumor growth. In addition, about 47% injection dose of anti-CXCR4-NaGdF4 NDs is found in the mouse urine at 24 h post-injection. These findings demonstrate that anti-CXCR4-NaGdF4 NDs enable to be used as renal clearable nanomedicine for biotherapy and MRI of breast cancer.

Coumarins and flavones from Ficus erecta and their anti-inflammatory activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Ficus erecta, a traditional Chinese She Ethnomedicine, has been historically utilized to treat various inflammatory conditions such as arthritis, nephritis, and osteoporosis. However, the underlying mechanisms accounting for its anti-inflammatory activity, as well as its active components, largely remain elusive. AIM OF THE STUDY: The purpose of this research was to investigate the chemical constituents of F. erecta that contribute to its anti-inflammatory effects. MATERIALS AND METHODS: Coumarins and flavones were obtained from the 95% EtOH extract of F. erecta using virous column chromatography and reversed-phase semipreparative HPLC. The structures of the new compounds were elucidated by extensive analysis of spectroscopic methods, including HRESIMS, 1D and 2D NMR spectra, and CD experiments. Cultured macrophage RAW264.7 cells were utilized for the anti-inflammatory experiments. MTT cell viability assay, Griess reagent method, ELISA, and Western blot experiments were employed to evaluate the anti-inflammatory activity and investigate the related mechanism. RESULTS: Four new (1-4) and eleven previously identified (5-16) coumarins, together with one new (17) and six known flavones (18-23) were isolated from the whole plant of F. erecta. Compounds 7 and 17 significantly reduced nitric oxide (NO) and prostaglandin E2 (PGE2) production without cytotoxic effects. Furthermore, compounds 7 and 17 reduced the production of proinflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6 in a concentration-dependent manner. Western blot analysis indicated that compounds 7 and 17 suppressed the expression of iNOS, COX-2, and p-IκBα in LPS-stimulated RAW264.7 macrophage cells. CONCLUSION: The current phytochemical investigations revealed that coumarins and flavones represent the primary chemical constituents of F. erecta. Compounds 7 and 17 exhibit potent anti-inflammatory properties, linked with the inhibition of NF-κB activation by preventing the degradation of IκBα phosphorylation. These compounds may serve as promising candidates for treating or preventing certain inflammatory diseases.

Fabrication of lignosulfonate-derived porous carbon via pH-tunable self-assembly strategy for efficient atrazine removal.

Herein, a straightforward protocol was developed for the one-pot synthesis of N-doped lignosulfonate-derived carbons (NLDCs) with a tunable porous structure using natural amino acids-templated self-assembly strategy. Specifically, histidine was employed as a template reagent, leading to the preparation of 10-NLDC-21 with remarkable characteristics, including the large specific surface area (SBET = 1844.5 m2/g), pore volume (Vmes = 1.22 cm3/g) and efficient adsorption for atrazine (ATZ) removal. The adsorption behavior of ATZ by NLDCs followed the Langmuir and pseudo-second-order models, suggesting a monolayer chemisorption nature of ATZ adsorption with the maximum adsorption capacity reached up to 265.77 mg/g. Furthermore, NLDCs exhibited excellent environmental adaptability and recycling performance. The robust affinity could be attributed to multi-interactions including pore filling, electrostatic attraction, hydrogen bonding and π-π stacking between the adsorbents and ATZ molecules. This approach offers a practical method for exploring innovative bio-carbon materials for sewage treatment.

Mechanical Properties and Pore Structure of Multiwalled Carbon Nanotube-Reinforced Reactive Powder Concrete for Three-Dimensional Printing Manufactured by Material Extrusion.

Three-dimensional (3D) concrete printing technology has been considered promising, attracting extensive attention in the engineering field. Multiwalled carbon nanotubes (MWCNTs) have been used as an additive to reinforce the cement-based material. However, the research on the 3D printed MWCNT-reinforced high-strength concrete is rare. This research is to study the mechanical properties and pore structure of MWCNT-reinforced reactive powder concrete (RPC) for 3D printing. In this research, the workability of the printed RPC mixture with MWCNTs was first tested to pass the criteria of 3D printing. Then, the enhancement effect of MWCNTs on the printed RPC was tested by mechanical properties after hardening. Meanwhile, strength-displacement curves were recorded. In addition, the pore structures of printed RPC were observed and analyzed by X-ray computed tomography (CT) images. The results show that 0.05 wt% MWCNTs have no effect on the workability of the printable RPC slurry. MWCNTs could enhance the mechanical properties of the printed RPC by filling the flaws inside the samples, increasing the viscosity of the RPC slurry and forming bridges between cracks. Besides, 0.05 wt% MWCNTs may cause the failure mode of the printed RPC from brittle failure to ductile failure. In addition, MWCNTs significantly reduced the porosity of the printed RPC by decreasing pores with a volume over 0.01 mm3. As CT images show, the interlayer zone (IZ) of the 3D printed RPC sample is prone to pores, and a higher volume fraction is evident. In particular, within the volume of IZs, the minimum volume fraction at the IZ of 3D printed RPC appears on sample with MWCNTs.

The association analysis of DNA methylation and transcriptomics identified BpCYCD3;2 as a participant in influencing cell division in autotetraploid birch (Betula pendula) leaves.

Polyploidization plays a crucial role in plant breeding and genetic improvement. Although the phenomenon of polyploidization affecting the area and number of plant epidermal pavement cells is well described, the underlying mechanism behind this phenomenon is still largely unknown. In this study, we found that the leaves of autotetraploid birch (Betula pendula) stopped cell division earlier and had a larger cell area. In addition, compared to diploids, tetraploids have a smaller stomatal density and fewer stomatal numbers. Genome-wide DNA methylation analysis revealed no significant difference in global DNA methylation levels between diploids and tetraploids. A total of 9154 differential methylation regions (DMRs) were identified between diploids and tetraploids, with CHH-type DMRs accounting for 91.73% of all types of DMRs. Further research has found that there are a total of 2105 differentially methylated genes (DMEGs) with CHH-type DMRs in birch. The GO functional enrichment results of DMEGs showed that differentially methylated genes were mainly involved in terms such as cellular process and metabolic process. The analysis of differentially methylated genes and differentially expressed genes suggests that hyper-methylation in the promoter region may inhibit the gene expression level of BpCYCD3;2 in tetraploids. To investigate the function of BpCYCD3;2 in birch, we obtained overexpression and repressed expression lines of BpCYCD3;2 through genetic transformation. The morphogenesis of both BpCYCD3;2-OE and BpCYCD3;2-RE lines was not affected. However, low expression of BpCYCD3;2 can lead to inhibition of cell division in leaves, and this inhibition of cell proliferation can be compensated for by an increase in cell size. Additionally, we found that the number and density of stomata in the BpCYCD3;2-RE lines were significantly reduced, consistent with the tetraploid. These data indicate that changes in cell division ability and stomatal changes in tetraploid birch can be partially attributed to low expression of the BpCYCD3;2 gene, which may be related to hyper-methylation in its promoter region. These results will provide new insights into the mechanism by which polyploidization affects plant development.

Preparation, characterization, and application of waterborne lignin-based epoxy resin as eco-friendly wood adhesive.

A series of novel waterborne lignin-based epoxy resin emulsions (WLEPs) were successfully synthesized, and then the WLEPs were cured with polyamide (PA) to give formaldehyde-free wood adhesives with high-performance. The chemical structures and properties of WLEP emulsions were determined. The effects of the emulsifiers on thermal and mechanical properties of the adhesives were investigated, and the potential application of WLEPs in the formulation of plywood were also evaluated. The results demonstrated that the WLEP dispersions presented excellent storage stability (>180 days) with their viscosities range from 110 mPa·s to 470 mPa·s and particle sizes in the range of 321-696 nm, which were beneficial for the fluidity and permeability of the wood adhesives. Furthermore, the thermal and mechanical properties of adhesives could be tuned effectively by controlling the length of PEG chains. The adhesive bearing PEG 6000 exhibited the highest tensile strength of 24.0 MPa and Young's modulus of 1439 MPa. Notably, the plywood prepared with the resulting adhesives displayed good bonding performance, especially water resistance, which were much higher than the national standard requirement for exterior-grade plywood type I. These results indicated that the WLEPs could be used as sustainable alternatives for traditional formaldehyde-based wood adhesives in practical applications.

BpWOX11 promotes adventitious root formation in Betula pendula.

Adventitious root formation is a key step in vegetative propagation via cuttings. It is crucial for establishing birch plantations and preserve birch varieties. Although previous studies have highlighted role of WOX11 in controlling adventitious root formation, no such study has been conducted in birch. Understanding the mechanism of adventitious root formation is essential for improvement of rooting or survival rate using stem cuttings in birch. In this study, we cloned BpWOX11 and produced BpWOX11 overexpression (OE) transgenic lines using the Agrobacterium-mediated plant transformation. OE lines exhibited early initiated adventitious root formation, leading to increase the rooting rate of stem cuttings plants. RNA sequencing analysis revealed that OE lines induced the gene expression related to expansin and cell division pathway, as well as defense and stress response genes. These may be important factors for the BpWOX11 gene to promote adventitious root formation in birch cuttings. The results of this study will help to further understand the molecular mechanisms controlling the formation of adventitious roots in birch.

Utilizing grayscale ultrasound-based radiomics nomogram for preoperative identification of triple negative breast cancer.

PURPOSE: This study aimed to develop a radiomics nomogram based on grayscale ultrasound (US) to distinguish triple-negative breast cancer (TNBC) from non-triple-negative breast cancer (NTNBC) prior to surgery. METHODS: A retrospective analysis of 454 breast carcinoma patients confirmed by pathology was conducted, with 317 patients in the training dataset (59 with TNBC) and 137 patients in the validation dataset (27 with TNBC). Clinical information, conventional US features, and radiomics features were collected, and the Radscore model was constructed after feature selection. Independent risk factors were identified using univariate and multivariate logistic regression analysis. The nomogram model was assessed using the receiver operating characteristic (ROC) curve analysis, calibration curve, decision curve analysis (DCA), net reclassification improvement (NRI) and integrated discrimination improvement (IDI). RESULTS: Tumor shape, margin, and calcification were independent risk factors in the clinical prediction model. Additionally, 16 radiomics features were selected to construct the Radscore model out of a total of 474 extracted features. The radiomics nomogram model, which incorporated tumor shape, margin, calcification, and Radscore, achieved an AUC value of 0.837 in the training dataset and 0.813 in the validation dataset, outperforming both the Radscore and clinical models in terms of predictive performance. The significant improvement of NRI and IDI indicated that the Radscore may be useful biomarkers for TNBC. CONCLUSION: The US-based radiomics nomogram showed satisfactory preoperative prediction of TNBC.

Development of an Intratumoral and Peritumoral Radiomics Nomogram Using Digital Breast Tomosynthesis for Preoperative Assessment of Lymphovascular Invasion in Invasive Breast Cancer.

RATIONALE AND OBJECTIVES: This study aimed to create a nomogram model that combines clinical factors with radiomics analysis of both intra- and peritumoral regions extracted from preoperative digital breast tomosynthesis (DBT) images, in order to develop a reliable method for predicting the lymphovascular invasion (LVI) status in invasive breast cancer (IBC) patients. MATERIALS AND METHODS: A total of 178 patients were randomly split into a training dataset (N = 124) and a validation dataset (N = 54). Comprehensive clinical data, encompassing DBT features, were gathered for all cases. Radiomics features were extracted and selected from intra- and peritumoral region to establish radiomics signature (Radscore). To construct the clinical model and nomogram model, univariate and multivariate logistic regression analyses were utilized to identify independent risk factors. To assess and validate these models, various analytical methods were employed, including receiver operating characteristic (ROC) curve analysis, calibration curve analysis, decision curve analysis (DCA), net reclassification improvement (NRI), and integrated discriminatory improvement (IDI). RESULTS: The clinical model is constructed based on two independent risk factors: tumor margin and the DBT-reported lymph node metastasis (DBT_reported_LNM). Incorporating Radscore_Combine (utilizing both intra- and peritumoral radiomics features), tumor margin, and DBT_reported_LNM into the nomogram achieved a reliable predictive performance, with area under the curve (AUC) values of 0.906 and 0.905 in both datasets, respectively. The significant improvement demonstrated by the NRI and IDI indicates that the Radscore_Combine could be a valuable biomarker for effectively predicting the status of LVI. CONCLUSION: The nomogram demonstrated a reliable ability to predict LVI in IBC patients.

Characterization of the F-Box Gene Family and Its Expression under Osmotic Stress in Birch.

The F-box gene family is abundant in plants and crucial for plant growth and development. However, two questions prevail: Which F-box genes are involved in regulating plant biological processes? How do these genes regulate such biological processes? In this study, we characterized the F-box family and identified 240 F-box genes in birch (Betula platyphylla Suk.) via HMMER analysis. According to the C-terminal conserved domains, the F-box members were divided into 10 subfamilies. Through phylogenetic analysis, the F-box proteins were clustered into eight evolutionary branches. Synteny analyses suggested that the birch F-box gene family exhibits tandem and segmental duplication events. GO annotation analysis revealed that BpF-box proteins respond to stimuli, and regulate the defense response. According to RNA-Seq analysis, we found that 11 differentially expressed genes (DEGs) are responsive to osmotic stress. We performed co-expression analysis on the representative genes, and GO enrichment analysis further revealed that representative plant genes participate in the regulation of hormones, growth, and development. Through qRT-PCR, we found that the representative BpF-box genes are mainly involved in hormone response signaling pathways. It appears that the F-box gene family plays a significant role in the regulation of birch osmotic stress responses through the regulation of different hormones. Our results provided novel insights into the biological function of BpF-box proteins.

Nifuratel reduces Salmonella survival in macrophages by extracellular and intracellular antibacterial activity.

Salmonella are intracellular bacterial pathogens for which, as with many of the other Enterobacteriaceae, antibiotic resistance is becoming an increasing problem. New antibiotics are being sought as recommended by the World Health Organization and other international institutions. These must be able to penetrate macrophages, and infect the major host cells and the Salmonella-containing vacuole. This study reports screening a small library of Food and Drug Administration (FDA)-approved drugs for their antibacterial effect in macrophages infected with a rapid-multiplying mutant of Salmonella Enteritidis. The most effective drug that was least toxic for macrophages was Nifuratel, a nitrofuran antibiotic already in use for parasitic infections. In mice, it provided 60% protection after oral infection with a lethal S. Enteritidis dose with reduced bacterial numbers in the tissues. It was effective against different serovars, including multidrug-resistant strains of Salmonella Typhimurium, and in macrophages from different host species and against Listeria monocytogenes and Shigella flexneri. It reduced IL-10 and STAT3 production in infected macrophages which should increase the inflammatory response against Salmonella. IMPORTANCE Salmonella can keep long-term persistence in host's macrophages to evade cellular immune defense and antibiotic attack and exit in some condition and reinfect to cause salmonellosis again. In addition to multidrug resistance, this infection circle causes Salmonella clearance difficult in the host, and so there is a great need for new antibacterial agents that reduce intramacrophage Salmonella survival to block endogenous Salmonella reinfection.

Functional study of BpCOI1 reveals its role in affecting disease resistance in birch.

Plants interact with biotic and abiotic environments. Some of these interactions are detrimental including herbivory consumption and infections by microbial pathogens. The COI1 (coronatine insensitive 1) protein is the master controller of JA-regulated plant responses and plays a regulatory role in the plant defense response. However, there is little information on COI1 function in birch (Betula platyphylla × Betula pendula). Herein, we studied the F-box protein BpCOI1 which is located in the nucleus. To validate the function of this protein, we developed transgenic birch plants with overexpression or repression of BpCOI1 gene. Growth traits, such as tree height, ground diameter, number of lateral branches, did not change significantly among transgenic lines. Alternaria alternata treatment experiments indicated that low expression of BpCOI1 reduced disease resistance in birch. Furthermore, our results showed that low expression of BpCOI1 significantly reduced the sensitivity of plants to exogenous MeJA. Co-expression analysis showed gene expression patterns with similar characteristics. These genes may be closely related in function, or members involved in the same signaling pathway or physiological process with BpCOI 1. The results of transcriptome sequencing and co-expression analysis showed that BpCOI1 affects plant defense against Alternaria alternata by regulating jasmonates. This study reveals the role of BpCOI1 in disease resistance and proposes the possibility of controlling diseases through molecular breeding in birch.

Heritable epigenetic modification of BpPIN1 is associated with leaf shapes in Betula pendula.

The new variety Betula pendula 'Dalecarlica', selected from Betula pendula, shows high ornamental value owing to its lobed leaf shape. In this study, to identify the genetic components of leaf shape formation, we performed bulked segregant analysis and molecular marker-based fine mapping to identify the causal gene responsible for lobed leaves in B. pendula 'Dalecarlica'. The most significant variations associated with leaf shape were identified within the gene BpPIN1 encoding a member of the PIN-FORMED family, responsible for the auxin efflux carrier. We further confirmed the hypomethylation at the promoter region promoting the expression level of BpPIN1, which causes stronger and longer veins and lobed leaf shape in B. pendula 'Dalecarlica'. These results indicated that DNA methylation at the BpPIN1 promoter region is associated with leaf shapes in B. pendula. Our findings revealed an epigenetic mechanism of BpPIN1 in the regulation of leaf shape in Betula  Linn. (birch), which could help in the molecular breeding of ornamental traits.

Radiomics nomogram based on digital breast tomosynthesis: preoperative evaluation of axillary lymph node metastasis in breast carcinoma.

PURPOSE: This study aimed to establish a radiomics nomogram model based on digital breast tomosynthesis (DBT) images, to predict the status of axillary lymph nodes (ALN) in patients with breast carcinoma. METHODS: The data of 120 patients with confirmed breast carcinoma, including 49 cases with axillary lymph node metastasis (ALNM), were retrospectively analyzed in this study. The dataset was randomly divided into a training group consisting of 84 patients (37 with ALNM) and a validation group comprising 36 patients (12 with ALNM). Clinical information was collected for all cases, and radiomics features were extracted from DBT images. Feature selection was performed to develop the Radscore model. Univariate and multivariate logistic regression analysis were employed to identify independent risk factors for constructing both the clinical model and nomogram model. To evaluate the performance of these models, receiver operating characteristic (ROC) curve analysis, calibration curve, decision curve analysis (DCA), net reclassification improvement (NRI), and integrated discriminatory improvement (IDI) were conducted. RESULTS: The clinical model identified tumor margin and DBT_reported_LNM as independent risk factors, while the Radscore model was constructed using 9 selected radiomics features. Incorporating tumor margin, DBT_reported_LNM, and Radscore, the radiomics nomogram model exhibited superior performance with AUC values of 0.933 and 0.920 in both datasets, respectively. The NRI and IDI showed a significant improvement, suggesting that the Radscore may serve as a useful biomarker for predicting ALN status. CONCLUSION: The radiomics nomogram based on DBT demonstrated effective preoperative prediction performance for ALNM in patients with breast cancer.