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Epithelial cells function as the primary line of defense against invading pathogens. However, bacterial pathogens possess the ability to compromise this barrier and facilitate the transmigration of bacteria. Nonetheless, the specific molecular mechanism employed by Mycobacterium tuberculosis (M.tb) in this process is not fully understood. Here, we investigated the role of Rv2569c in M.tb translocation by assessing its ability to cleave E-cadherin, a crucial component of cell-cell adhesion junctions that are disrupted during bacterial invasion. By utilizing recombinant Rv2569c expressed in Escherichia coli and subsequently purified through affinity chromatography, we demonstrated that Rv2569c exhibited cell wall-associated serine protease activity. Furthermore, Rv2569c was capable of degrading a range of protein substrates, including casein, fibrinogen, fibronectin, and E-cadherin. We also determined that the optimal conditions for the protease activity of Rv2569c occurred at a temperature of 37°C and a pH of 9.0, in the presence of MgCl2. To investigate the function of Rv2569c in M.tb, a deletion mutant of Rv2569c and its complemented strains were generated and used to infect A549 cells and mice. The results of the A549-cell infection experiments revealed that Rv2569c had the ability to cleave E-cadherin and facilitate the transmigration of M.tb through polarized A549 epithelial cell layers. Furthermore, in vivo infection assays demonstrated that Rv2569c could disrupt E-cadherin, enhance the colonization of M.tb, and induce pathological damage in the lungs of C57BL/6 mice. Collectively, these results strongly suggest that M.tb employs the serine protease Rv2569c to disrupt epithelial defenses and facilitate its systemic dissemination by crossing the epithelial barrier.
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Proteínas de Bactérias , Caderinas , Células Epiteliais , Mycobacterium tuberculosis , Serina Proteases , Caderinas/metabolismo , Mycobacterium tuberculosis/patogenicidade , Mycobacterium tuberculosis/metabolismo , Animais , Humanos , Camundongos , Serina Proteases/metabolismo , Serina Proteases/genética , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Células A549 , Tuberculose/microbiologia , Tuberculose/metabolismo , FemininoRESUMO
AIMS/HYPOTHESIS: Mutations in Isl1, encoding the insulin enhancer-binding protein islet-1 (ISL1), may contribute to attenuated insulin secretion in type 2 diabetes mellitus. We made an Isl1E283D mouse model to investigate the disease-causing mechanism of diabetes mellitus. METHODS: The ISL1E283D mutation (c. 849A>T) was identified by whole exome sequencing on an early-onset type 2 diabetes family and then the Isl1E283D knockin (KI) mouse model was created and an IPGTT and IPITT were conducted. Glucose-stimulated insulin secretion (GSIS), expression of Ins2 and other ISL1 target genes and interacting proteins were evaluated in isolated pancreas islets. Transcriptional activity of Isl1E283D was evaluated by cell-based luciferase reporter assay and electrophoretic mobility shift assay, and the expression levels of Ins2 driven by Isl1 wild-type (Isl1WT) and Isl1E283D mutation in rat INS-1 cells were determined by RT-PCR and western blotting. RESULTS: Impaired GSIS and elevated glucose level were observed in Isl1E283D KI mice while expression of Ins2 and other ISL1 target genes Mafa, Pdx1, Slc2a2 and the interacting protein NeuroD1 were downregulated in isolated islets. Transcriptional activity of the Isl1E283D mutation for Ins2 was reduced by 59.3%, and resulted in a marked downregulation of Ins2 expression when it was overexpressed in INS-1 cells, while overexpression of Isl1WT led to an upregulation of Ins2 expression. CONCLUSIONS/INTERPRETATION: Isl1E283D mutation reduces insulin expression and secretion by regulating insulin and other target genes, as well as its interacting proteins such as NeuroD1, leading to the development of glucose intolerance in the KI mice, which recapitulated the human diabetic phenotype. This study identified and highlighted the Isl1E283D mutation as a novel causative factor for type 2 diabetes, and suggested that targeting transcription factor ISL1 could offer an innovative avenue for the precise treatment of human type 2 diabetes.
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Diabetes Mellitus Tipo 2 , Proteínas com Homeodomínio LIM , Mutação de Sentido Incorreto , Fatores de Transcrição , Animais , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/metabolismo , Proteínas com Homeodomínio LIM/genética , Proteínas com Homeodomínio LIM/metabolismo , Camundongos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Humanos , Masculino , Insulina/metabolismo , Feminino , Ratos , Secreção de Insulina/genética , Ilhotas Pancreáticas/metabolismoRESUMO
Amylose biosynthesis is strictly associated with granule-bound starch synthase I (GBSSI) encoded by the Waxy gene. Mutagenesis of single bases in the Waxy gene, which induced by CRISPR/Cas9 genome editing, caused absence of intact GBSSI protein in grain of the edited line. The amylose and amylopectin contents of waxy mutants were zero and 31.73%, while those in the wild type were 33.50% and 39.00%, respectively. The absence of GBSSI protein led to increase in soluble sugar content to 37.30% compared with only 10.0% in the wild type. Sucrose and ß-glucan, were 39.16% and 35.40% higher in waxy mutants than in the wild type, respectively. Transcriptome analysis identified differences between the wild type and waxy mutants that could partly explain the reduction in amylose and amylopectin contents and the increase in soluble sugar, sucrose and ß-glucan contents. This waxy flour, which showed lower final viscosity and setback, and higher breakdown, could provide more option for food processing.
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Amilose , Edição de Genes , Hordeum , Proteínas de Plantas , Sintase do Amido , Amilose/metabolismo , Hordeum/genética , Hordeum/metabolismo , Edição de Genes/métodos , Sintase do Amido/genética , Sintase do Amido/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sistemas CRISPR-Cas , Amilopectina/metabolismo , Sacarose/metabolismo , Açúcares/metabolismo , Regulação da Expressão Gênica de Plantas , Mutação , beta-Glucanas/metabolismo , Plantas Geneticamente Modificadas , SolubilidadeRESUMO
Room-temperature gallium-based liquid metals (RT-GaLMs) have garnered significant interest recently owing to their extraordinary combination of fluidity, conductivity, stretchability, self-healing performance, and biocompatibility. They are ideal materials for the manufacture of flexible electronics. By changing the composition and oxidation of RT-GaLMs, physicochemical characteristics of the liquid metal can be adjusted, especially the regulation of rheological, wetting, and adhesion properties. This review highlights the advancements in the liquid metals used in flexible electronics. Meanwhile related characteristics of RT-GaLMs and underlying principles governing their processing and applications for flexible electronics are elucidated. Finally, the diverse applications of RT-GaLMs in self-healing circuits, flexible sensors, energy harvesting devices, and epidermal electronics, are explored. Additionally, the challenges hindering the progress of RT-GaLMs are discussed, while proposing future research directions and potential applications in this emerging field. By presenting a concise and critical analysis, this paper contributes to the advancement of RT-GaLMs as an advanced material applicable for the new generation of flexible electronics.
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Copper-nitrogen-doped carbon-based nanocatalysts (Cu-NCs), containing atomically dispersed Cu-NxC4- x sites, are efficient in boosting the Fenton-like reaction. However, the mechanisms of the Fenton-like reaction, including the pH effect on the products and the effect of the coordination environment on catalytic activity, remain controversial, restricting the development of Cu-NCs. Cu-NCs are experimentally synthesized with Cu-N4 sites and prove that the Fenton-like reaction generates mainly hydroxyl radicals (·OH) in the acidic but ·OH and superoxide radicals (·O2 -) in the neutral. The density functional theory (DFT) calculations reveal that the catalytic activity of Cu-NCs in the Fenton-like reaction is associated with the adsorption strength of ·OH at the Cu site. Further investigation of the effect of the coordination environment of Cu-NCs indicates that the Cu-N2C2 site, which can enhance the ·OH adsorption strength, is an ideal catalyst site for the Fenton-like reaction. These results open the way to facilitating the catalytic activity of Cu-NCs in the Fenton-like reaction.
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Amplifying oxidative stress within tumor cells can effectively inhibit the growth and metastasis of triple-negative breast cancer (TNBC). Therefore, the development of innovative nanomedicines that can effectively disrupt the redox balance represents a promising yet challenging therapeutic strategy for TNBC. In this study, an oxidative stress amplifier, denoted as PBCH, comprising PdAg mesoporous nanozyme and a CaP mineralized layer, loaded with GSH inhibitor L-buthionine sulfoximine (BSO), and further surface-modified with hyaluronic acid that can target CD44, is introduced. In the acidic tumor microenvironment, Ca2+ is initially released, thereby leading to mitochondrial dysfunction and eventually triggering apoptosis. Additionally, BSO suppresses the synthesis of intracellular reduced GSH and further amplifies the level of oxidative stress in cancer cells. Furthermore, PdAg nanozyme can be activated by near-infrared light to induce photothermal and photodynamic effects, causing a burst of ROS and simultaneously promoting cell apoptosis via provoking immunogenic cell death. The high-performance therapeutic effects of PBCH, based on the synergistic effect of aforementioned multiple oxidative damage and photothermal ablation, are validated in TNBC cells and animal models, declaring its potential as a safe and effective anti-tumor agent. The proposed approach offers new perspectives for precise and efficient treatment of TNBC.
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Atoms doping is a practical approach to modulate the physicochemical properties of carbon dots (CDs) and thus has garnered increasing attention in recent years. Compared to non-metal atoms, transition metal atoms (TMAs) possess more unoccupied orbitals and larger atomic radii. TMAs doping can significantly alter the electronic structure of CDs and bestow them with new intrinsic characteristics. TMAs-doped CDs have exhibited widespread application potential as a new class of single-atom-based nanomaterials. However, challenges remain for the successful preparation and precise design of TMAs-doped CDs. The key to successfully preparing TMA-doped CDs lies in anchoring TMAs to the carbon precursors before the reaction. Herein, taking the formation mechanism of TMAs-doped CDs as a starting point, we systematically summarized the ligands employed for synthesizing TMAs-doped CDs and proposed the synthetic strategy involving multiple ligands. Additionally, we summarize the functional properties imparted to CDs by different TMA dopants to guide the design of TMA-doped CDs with different functional characteristics. Finally, we describe the bottlenecks TMAs-doped CDs face and provide an outlook on their future development.
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Nanoestruturas , Elementos de Transição , Carbono , EletrônicaRESUMO
BACKGROUND: Cryptosporidium is a highly pathogenic parasite responsible for diarrhea in children worldwide. Here, the epidemiological status and genetic characteristics of Cryptosporidium in children with or without diarrhea were investigated with tracking of potential sources in Wenzhou City, China. METHODS: A total of 1032 children were recruited, 684 of whom had diarrhea and 348 without, from Yuying Children's Hospital in Wenzhou, China. Samples of stool were collected from each participant, followed by extraction of DNA, genotyping, and molecular identification of Cryptosporidium species and subtypes. RESULTS: Twenty-two of the 1032 (2.1%) children were infected with Cryptosporidium spp. with 2.5% (17/684) and 1.4% (5/348) in diarrhoeic and asymptomatic children, respectively. Four Cryptosporidium species were identified, including C. parvum (68.2%; 15/22), C. felis (13.6%; 3/22), C. viatorum (9.1%; 2/22), and C. baileyi (9.1%; 2/22). Two C. parvum subtypes named IIdA19G1 (n = 14) and IInA10 (n = 1), and one each of C. felis (XIXa) and C. viatorum (XVaA3g) subtype was found as well. CONCLUSIONS: This is the first research that identified Cryptosporidium in children of Wenzhou, China, using PCR. Identification of zoonotic C. parvum, C. felis, C. viatorum, and their subtypes indicate potential cross-species transmission of Cryptosporidium between children and animals. Additionally, the presence of C. baileyi in children suggests that this species has a wider host range than previously believed and that it possesses the capacity to infect humans.
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Criptosporidiose , Cryptosporidium , Criança , Animais , Humanos , Cryptosporidium/genética , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , Diarreia/epidemiologia , China/epidemiologia , Fezes/parasitologia , Genótipo , ProbabilidadeRESUMO
Through extensive density functional theory (DFT) calculations, our investigation delves into the stability, electrical characteristics, and magnetic behavior of monolayers (MLs) of MSi2N4. Computational analyses indicate intrinsic antiferromagnetic (AFM) orders within the MSi2N4 MLs, as a result of direct exchange interactions among transition metal (M) atoms. We further find that CrSi2N4 and CoSi2N4 MLs with primitive cells (pcells) exhibit half-metallic properties, with respective spin-ß electron gaps of 3.661 and 2.021 eV. In contrast, MnSi2N4 and FeSi2N4 MLs with pcells act as semiconductors, having energy gaps of 0.427 and 0.282 eV, respectively. When the SOC is considered, the CrSi2N4, MnSi2N4 and FeSi2N4 MLs are metals, while the CoSi2N4 ML is a semiconductor. Our findings imply the dynamics and thermodynamic stability of MSi2N4 MLs. We have also explored the influence of carrier doping on the electromagnetic attributes of MSi2N4 MLs. Interestingly, charge doping could transform CrSi2N4, MnSi2N4, and CoSi2N4 MLs from their original AFM state into a ferromagnetic (FM) order. Moreover, carrier doping transformed CrSi2N4 and CoSi2N4 MLs from spin-polarized metals to half-metals (HMs). It is of particular note that doping of CrSi2N4 MLs with +0.9 e per pcell or more holes caused a switch in the easy axis (EA) to the [001] axis. The demonstrated intrinsic AFM order, excellent thermodynamic and kinetic stability, adjustable magnetism, and half-metallicity of the MSi2N4 family suggest its promising potential for applications in the realm of spintronics.
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Varieties of RNA modification form the epitranscriptome for post-transcriptional regulation. 5-Methylcytosine (5-mC) is a sparse RNA modification in messenger RNA (mRNA) under physiological conditions. The function of RNA 5-hydroxymethylcytosine (5-hmC) oxidized by ten-eleven translocation (Tet) proteins in Drosophila has been revealed more recently. However, the turnover and function of 5-mC in mammalian mRNA have been largely unknown. Tet2 suppresses myeloid malignancies mostly in an enzymatic activity-dependent manner, and is important in resolving inflammatory response in an enzymatic activity-independent way. Myelopoiesis is a common host immune response in acute and chronic infections; however, its epigenetic mechanism needs to be identified. Here we demonstrate that Tet2 promotes infection-induced myelopoiesis in an mRNA oxidation-dependent manner through Adar1-mediated repression of Socs3 expression at the post-transcription level. Tet2 promotes both abdominal sepsis-induced emergency myelopoiesis and parasite-induced mast cell expansion through decreasing mRNA levels of Socs3, a key negative regulator of the JAK-STAT pathway that is critical for cytokine-induced myelopoiesis. Tet2 represses Socs3 expression through Adar1, which binds and destabilizes Socs3 mRNA in a RNA editing-independent manner. For the underlying mechanism of Tet2 regulation at the mRNA level, Tet2 mediates oxidation of 5-mC in mRNA. Tet2 deficiency leads to the transcriptome-wide appearance of methylated cytosines, including ones in the 3' untranslated region of Socs3, which influences double-stranded RNA formation for Adar1 binding, probably through cytosine methylation-specific readers, such as RNA helicases. Our study reveals a previously unknown regulatory role of Tet2 at the epitranscriptomic level, promoting myelopoiesis during infection in the mammalian system by decreasing 5-mCs in mRNAs. Moreover, the inhibitory function of cytosine methylation on double-stranded RNA formation and Adar1 binding in mRNA reveals its new physiological role in the mammalian system.
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Proteínas de Ligação a DNA/metabolismo , Mielopoese , Proteínas Proto-Oncogênicas/metabolismo , RNA Mensageiro/química , RNA Mensageiro/metabolismo , Sepse/genética , Sepse/microbiologia , Regiões 3' não Traduzidas/genética , 5-Metilcitosina/análogos & derivados , 5-Metilcitosina/metabolismo , Adenosina Desaminase/metabolismo , Animais , Células da Medula Óssea/imunologia , Proteínas de Ligação a DNA/deficiência , Dioxigenases , Epigênese Genética , Feminino , Regulação da Expressão Gênica , Imunidade Inata , Camundongos , Mielopoese/genética , Conformação de Ácido Nucleico , Oxirredução , Proteínas Proto-Oncogênicas/deficiência , RNA de Cadeia Dupla/química , RNA de Cadeia Dupla/genética , RNA de Cadeia Dupla/metabolismo , RNA Mensageiro/genética , Proteína 3 Supressora da Sinalização de Citocinas/genética , Transcriptoma/genéticaRESUMO
BACKGROUND: The environmental pollution and ecological risks caused by the widespread use of antibiotics have attracted attention in recent years. Biochar materials have a rich pore diameter and can effectively adsorb pollutants from wastewater. However, biochar will experience high temperatures, freezing and thawing in nature, affecting its physicochemical properties and adsorption capacity. Three types of aged biochar were prepared by artificial simulated aging using soybean straw as raw material. The aged biochar's elemental composition and functional group species were investigated by characterization analysis, and their adsorption kinetics and adsorption isotherms were studied. RESULTS: The specific surface area and pore size of the three aged biochars were lower than those of fresh biochars. The increased number of oxygen-containing functional groups of the aged biochars formed a water cluster interaction with norfloxacin (NOR), which was unfavorable to the adsorption of NOR. The adsorption mechanism of biochar on NOR comprises pore filling, electrostatic interaction, ion exchange and complexation. CONCLUSION: The adsorption of NOR on biochar before and after aging was spontaneous and was described by quasi-second kinetics and the Langmuir equation. Different aging methods influenced the physicochemical properties and adsorption performance of biochar, and the adsorption capacity of biochar was significantly reduced after aging. Therefore, the influence of climatic factors needs to be considered when using biochar to remove target pollutants. © 2023 Society of Chemical Industry.
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Poluentes Ambientais , Poluentes Químicos da Água , Adsorção , Águas Residuárias , Glycine max , Antibacterianos , Carvão Vegetal/química , Cinética , Poluentes Químicos da Água/químicaRESUMO
To fully understand whether Saposhnikoviae Radix polysaccharides(SP) can be metabolized in gastric fluid and the meta-bolic behavior, this study systematically analyzed the metabolites in simulated gastric fluid of SP by high-performance liquid chromatography-ion trap time-of-flight mass spectrometry(HPLC-IT-TOF-MS) technology in combination with zebrafish immune activity evaluation. Based on the obtained accurate relative molecular mass, chromatographic retention behavior, MS fragmentation patterns, refe-rence standards, and relevant literature reports, 19 metabolites were analyzed and identified. Among them, five monosaccharides and 14 oligosaccharides were generated as metabolites. Several reducing sugars, including mannose, glucose, rhamnose, and xylose, were accurately identified in the gastric fluid metabolites. Zebrafish pharmacological evaluation results indicated that SP maintained good immune activity after gastric fluid metabolism, with the most significant increase in immune cell density observed at W3(simulated gastric fluid metabolism for 2 hours). Among the gastric fluid metabolites, M1 and M3(Hex-Hex-Man) may be most closely related to pharmacological activity and could be further studied as potential active fragments.
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Medicamentos de Ervas Chinesas , Peixe-Zebra , Humanos , Animais , Cromatografia Líquida de Alta Pressão/métodos , Raízes de Plantas/química , Polissacarídeos/análise , Medicamentos de Ervas Chinesas/químicaRESUMO
Lung cancer remains the leading cause of cancer-related death due to poor treatment responses and resistance arising from tumour heterogeneity. Here, we show that adverse prognosis associated with epigenetic silencing of the tumour suppressor RASSF1A is due to increased deposition of extracellular matrix (ECM), tumour stiffness and metastatic dissemination in vitro and in vivo. We find that lung cancer cells with RASSF1A promoter methylation display constitutive nuclear YAP1 accumulation and expression of prolyl 4-hydroxylase alpha-2 (P4HA2) which increases collagen deposition. Furthermore, we identify that elevated collagen creates a stiff ECM which in turn triggers cancer stem-like programming and metastatic dissemination in vivo. Re-expression of RASSF1A or inhibition of P4HA2 activity reverses these effects and increases markers of lung differentiation (TTF-1 and Mucin 5B). Our study identifies RASSF1A as a clinical biomarker associated with mechanical properties of ECM which increases the levels of cancer stemness and risk of metastatic progression in lung adenocarcinoma. Moreover, we highlight P4HA2 as a potential target for uncoupling ECM signals that support cancer stemness.
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Adenocarcinoma de Pulmão/patologia , Metilação de DNA , Neoplasias Pulmonares/patologia , Células-Tronco Neoplásicas/metabolismo , Proteínas Supressoras de Tumor/genética , Regulação para Cima , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Adenocarcinoma de Pulmão/genética , Adenocarcinoma de Pulmão/metabolismo , Animais , Linhagem Celular Tumoral , Progressão da Doença , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Camundongos , Metástase Neoplásica , Transplante de Neoplasias , Prolil Hidroxilases/metabolismo , Regiões Promotoras Genéticas , Fatores de Transcrição/metabolismo , Proteínas de Sinalização YAPRESUMO
Genome editing has become more and more popular in animal and plant systems following the emergence of CRISPR/Cas9 technology. However, target sequence modification by CRISPR/Cas9 has not been reported in the plant mitochondrial genome, mtDNA. In plants, a type of male sterility known as cytoplasmic male sterility (CMS) has been associated with certain mitochondrial genes, but few genes have been confirmed by direct mitochondrial gene-targeted modifications. Here, the CMS-associated gene (mtatp9) in tobacco was cleaved using mitoCRISPR/Cas9 with a mitochondrial localization signal. The male-sterile mutant, with aborted stamens, exhibited only 70% of the mtDNA copy number of the wild type and exhibited an altered percentage of heteroplasmic mtatp9 alleles; otherwise, the seed setting rate of the mutant flowers was zero. Transcriptomic analyses showed that glycolysis, tricarboxylic acid cycle metabolism and the oxidative phosphorylation pathway, which are all related to aerobic respiration, were inhibited in stamens of the male-sterile gene-edited mutant. In addition, overexpression of the synonymous mutations dsmtatp9 could restore fertility to the male-sterile mutant. Our results strongly suggest that mutation of mtatp9 causes CMS and that mitoCRISPR/Cas9 can be used to modify the mitochondrial genome of plants.
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Genoma Mitocondrial , Infertilidade Masculina , Animais , Masculino , Humanos , Edição de Genes , Sistemas CRISPR-Cas , Nicotiana/genética , DNA Mitocondrial/genética , Infertilidade Masculina/genéticaRESUMO
Hypoxia is a key impediment encountered in the treatment of most solid tumors, leading to immune escape and therapeutic resistance. Perfluorocarbons (PFCs) have a unique electrical structure and are characterized by a high solubility for gases. PFC-based oxygen carriers have been evaluated for their ability to deliver oxygen effectively to hypoxic tissues, and significant clinical translation has been demonstrated. And due to the unique acoustic activity, PFCs have been employed to stabilize the injection of gas microbubbles (MBs) as clinical ultrasonography contrast agents. In contrast, the ultrasound and photothermally activatable PFC phase-shift nanodroplets (P-SNDs) represent a novel alternative to ultrasound imaging and hypoxia improvement. The PFC-based oxygen carriers may be utilized to improve the efficacy of cancer treatments based on synergistic radiotherapy (RT), chemotherapy (CMT), and photodynamic therapy (PDT) to reshape the tumor microenvironment through synergistic immunotherapy (IMT) and to achieve precise tumor diagnosis using acoustic imaging. This review described the characteristics of PFCs to provide an update on the design of PFC delivery systems used for oxygen delivery and ultrasound imaging to facilitate the treatment and diagnosis of tumors. The objective was to contribute to overcoming the obstacles encountered during PFC research and provide the developing prospects.
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Fluorocarbonos , Neoplasias , Humanos , Medicina de Precisão , Fluorocarbonos/química , Neoplasias/diagnóstico por imagem , Neoplasias/tratamento farmacológico , Oxigênio , Hipóxia , Microambiente TumoralRESUMO
The combination of vascular endothelial growth factor (VEGF) inhibitors and tyrosine kinase inhibitors (TKIs) is newly available for molecular targeted therapy against non-small cell lung cancer (NSCLC) in clinic. However, the therapeutic benefits remain unsatisfying due to the poor drug delivery to targets of interest. In this study, we developed bevacizumab-coated gefitinib-loaded nanoparticles (BCGN) with dual-responsive drug release for inhibiting tumor angiogenesis and phosphorylation of epidermal growth factor receptor (EGFR). Through an exogenous corona strategy, bevacizumab is easily coated on gefitinib-loaded nanoparticles via electrostatic interaction. After intravenous injection, BCGN are efficiently accumulated in NSCLC tumors as confirmed by dual-model imaging. Bevacizumab is released from BCGN upon oxidation in tumor microenvironment, whereas gefitinib is released after being internalized by tumor cells and disassembled in reduction cytoplasm. The dual-responsive release of bevacizumab and gefitinib significantly inhibits tumor growth in both A549 and HCC827 human NSCLC models. Our approach provides a promising strategy to improve combinational molecular targeted therapy of NSCLC with precisely controlled drug release.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Humanos , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Gefitinibe , Bevacizumab/uso terapêutico , Neoplasias Pulmonares/patologia , Fator A de Crescimento do Endotélio Vascular , Terapia de Alvo Molecular , Quinazolinas/farmacologia , Resistencia a Medicamentos Antineoplásicos , Linhagem Celular Tumoral , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , Microambiente TumoralRESUMO
Blue honeysuckle is emerging as a popular edible fruit and folk medicine. However, from June to August 2021, a serious leaf-spot disease affected the yield and quality of blue honeysuckle in Harbin, Heilongjiang Province, China; the species and characteristics of the pathogens responsible for the disease are unknown. In this study, 30 fungal isolates were obtained from infected blue honeysuckle leaves, identified as Alternaria tenuissima based on morphological and molecular characteristics and phylogenetic analyses. To the best of our knowledge, this is one of the first studies to identify A. tenuissima as the causal agent of blue honeysuckle leaf spots in China. Pathogenicity tests of the isolates revealed that most isolates exhibited moderately pathogenic. All blue honeysuckle cultivars tested were found to be susceptible to 30 A. tenuissima isolates. In addition, elder, Dahurian rose fruit, sea-buckthorn, rowan, hawthorn, bird cherry, and sorb could be infected by A. tenuissima isolates, while European cranberry bush and nanking cherry were not infected. A. tenuissima isolates were highly sensitive to prochloraz (EC50 ≤ 0.50 µg·ml-1) with 86.21% efficacy at 400 µg·ml-1 in the field trials. Therefore, the application of rotation and chemical fungicides are considered to control the disease-causing leaf spots in blue honeysuckle. These results provide a basis for controlling A. tenuissima in blue honeysuckle in China.
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Saposhnikovia divaricata is an authentic Chinese herbal medicine in Northeast China named Guanfangfeng, which is made from very high quality plants for sufficient efficacy. However, leaf spot causes a very large reduction in the yield and quality of S. divaricata in Shuangyashan (126°54'E, 45°81'N), Northeast China. A total of 18 isolates were isolated from the diseased leaves of S. divaricata, following Koch's postulates, and identified as Fusarium acuminatum based on morphological, molecular biological, and phylogenetic tree analyses. To the authors' knowledge, this is the first report of F. acuminatum causing S. divaricata leaf spot in China. F. acuminatum infected perilla and mung beans, but not foxtail millet, peanuts, wheat, peas, rye, red beans, or sorghum. Susceptibility assessment of F. acuminatum to fungicides using the mycelial growth rate method showed that isolates of F. acuminatum were most sensitive to prochloraz, with EC50 values of 0.0005413-0.0009523 µg·ml-1. In the two field experiments, the average control efficacy of prochloraz at 0.450 g/l on S. divaricata leaf spot caused by F. acuminatum was 75.42%. Therefore, non-host plant rotation or intercropping with suitable chemical fungicides may be used to control S. divaricata leaf spot. This study's results provide a theoretical basis for controlling S. divaricata leaf spot and will facilitate the development of effective disease management programs.
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Nitrogen (N) and phosphorus (P) are two important nutrient elements that limit the growth of plants and microorganisms. The effect of the N supply on soil P cycling and its mechanism remain poorly known. Here, we characterized the effects of different N application rates on soil P availability, the abundances of P-cycling functional genes, and microbial communities involved in P-cycling following the application of N for 13 years in a tea plantation. Soil available P (AP) decreased significantly under N application. The opposite pattern was observed for the activity of soil phosphatases including alkaline (ALP) and acid phosphatase (ACP). Furthermore, N addition increased the abundance of ppa but decreased the abundance of phoD in soil. Both ppa- and phoD-harboring communities varied with N application levels. Redundancy analysis (RDA) showed that soil pH was a key variable modulating ppa-harboring and phoD-harboring microbial communities. Partial least squares path modeling (PLS-PM) revealed that long-term N application indirectly reduced soil P availability by altering the abundances of phoD-harboring biomarker taxa. Overall, our findings indicated that N-induced reductions in AP increased microbial competition for P by selecting microbes with P uptake and starvation response genes or those with phosphatases in tea plantation system. This suggests that tea plantations should be periodically supplemented with P under N application, especially under high N application levels.
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Camellia sinensis , Microbiota , Solo/química , Fósforo/análise , Nitrogênio/análise , Microbiologia do Solo , Monoéster Fosfórico Hidrolases/farmacologia , CháRESUMO
Heteroatom doping, particularly with nonmetallic atoms such as N, P, and S, has proven to be an effective strategy for modulating the fluorescent properties of carbon dots (CDs). However, there are few reports on the regulation of the photoluminescence of CDs by transition-metal doping. In this work, nickel-doped CDs (Ni-CDs) were fabricated using the hydrothermal approach. Ni atoms were incorporated into the sp2 domains of the CDs through Ni-N bonds, resulting in an increased degree of graphitization of the Ni-CDs. Additionally, Ni-atom doping served to shorten the electron transition and recombination lifetimes, and suppress the nonradiative recombination process, resulting in an absolute fluorescence quantum yield of 54.7% for the Ni-CDs. Meanwhile, the as-prepared Ni-CDs exhibited excellent biocompatibility and were utilized for fluorescent bioimaging of HeLa cells. Subsequently, the Ni-CDs were employed as fluorescent anticounterfeiting inks for the successful encryption of two-dimensional barcodes. Our work demonstrates a novel heteroatom doping strategy for the synthesis of highly fluorescence-emitting CDs.