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Gene transfection, defined by the delivery of nucleic acids into cellular compartments, stands as a crucial procedure in gene therapy. While branched polyethylenimine (PEI) is widely regarded as the "gold standard" for nonviral vectors, its cationic nature presents several issues, including nonspecific protein adsorption and notable cytotoxicity. Additionally, it often fails to achieve high transfection efficiency, particularly with hard-to-transfect cell types. To overcome these challenges associated with PEI as a vector for plasmid DNA (pDNA), the photothermal agent indocyanine green (ICG) is integrated with PEI and pDNA to form the PEI/ICG/pDNA (PI/pDNA) complex for more efficient and safer gene transfection. The negatively charged ICG serves a dual purpose: neutralizing PEI's excessive positive charges to reduce cytotoxicity and, under near-infrared irradiation, inducing local heating that enhances cell membrane permeability, thus facilitating the uptake of PI/pDNA complexes to boost transfection efficiency. Using pDNA encoding vascular endothelial growth factor as a model, our system shows enhanced transfection efficiency in vitro for hard-to-transfect endothelial cells, leading to improved cell proliferation and migration. Furthermore, in vivo studies reveal the therapeutic potential of this system in accelerating the healing of infected wounds by promoting angiogenesis and reducing inflammation. This approach offers a straightforward and effective method for gene transfection, showing potentials for tissue engineering and cell-based therapies.
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The regulation of fruit development is a complex process and a core issue in the fruit tree industry. To investigate the role of PbGIF1 in pear fruit development, we identified a transcription factor PbbHLH137 that regulates pear (Pyrus bretschneideri) fruit development by screening a yeast library constructed from fruit cDNA. Yeast two-hybrid (Y2H), bimolecular fluorescence complementation (BiFC), and split luciferase complementation (split-LUC) assays were performed to confirm the PbbHLH137-PbGIF1 interaction. By tracing the complete fruit development process, we found that PbbHLH137 expression was closely related to fruit size and highly involved at the late pear fruit development stage. Transgenic experiments showed that heterologous expression of PbbHLH137 or PbGIF1 promoted fruit enlargement. PbbHLH137 promoted mainly the expansion of fruit cell volume, whereas PbGIF1 mainly increased the number of cells. Further LUC experiments demonstrated that PbGIF1 promoted the transcriptional activation ability of PbbHLH137. Our work identified PbbHLH137 as a transcription factor that regulates fruit development, and showed that PbGIF1 played an ongoing role during fruit development, making it a candidate gene for genetic improvement of pear fruit development.
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Frutas , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Pyrus , Fatores de Transcrição , Pyrus/genética , Pyrus/crescimento & desenvolvimento , Pyrus/metabolismo , Frutas/crescimento & desenvolvimento , Frutas/genética , Frutas/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Plantas Geneticamente Modificadas , Técnicas do Sistema de Duplo-HíbridoRESUMO
To improve the color stability of anthocyanins (ACNs) in blueberry fermented beverage, the intermolecular copigmentation between ACNs and 3 different phenolic compounds, including (-)-epigallocatechin gallate (EGCG), ferulic acid (FA), and gallic acid (GA) as copigments, was compared in the model and the real blueberry fermented beverage, respectively. The copigmented ACNs by EGCG presented a high absorbance (0.34 a.u.) and redness (27.09 ± 0.17) in the model blueberry fermented beverage. The copigmentation by the participation of the 3 different phenolic compounds showed all a spontaneous exothermic reaction, and the Gibbs free energy (ΔG°) of the system was lowest (-5.90 kJ/mol) using EGCG as copigment. Furthermore, the molecular docking model verified that binary complexes formed between ACNs and copigments by hydrogen bonds and π-π stacking. There was a high absorbance (1.02 a.u.), percentage polymeric color (PC%, 68.3 %), and good color saturation (C*ab, 43.28) in the real blueberry fermented beverage aged for 90 days, and more malvidin-3-O-glucoside had been preserved in the wine using EGCG as copigment. This finding may guide future industrial production of blueberry fermented beverage with improved color.
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Antocianinas , Mirtilos Azuis (Planta) , Cor , Ácidos Cumáricos , Fermentação , Ácido Gálico , Simulação de Acoplamento Molecular , Fenóis , Antocianinas/química , Mirtilos Azuis (Planta)/química , Ácidos Cumáricos/química , Ácido Gálico/química , Ácido Gálico/análogos & derivados , Fenóis/análise , Fenóis/química , Catequina/química , Catequina/análogos & derivados , Sucos de Frutas e Vegetais/análise , Frutas/químicaRESUMO
PURPOSE: This study aims to investigate the effects of chest wall bolus in intensity-modulated radiotherapy (IMRT) technology on clinical outcomes for post-mastectomy breast cancer patients. MATERIALS AND METHODS: This retrospective study included patients with invasive carcinoma ((y)pT0-4, (y)pN0-3) who received photon IMRT after mastectomy at the Affiliated Hospital of Qingdao University from 2014 to 2019. The patients were divided into two groups based on whether they received daily bolus application or not, and the baseline characteristics were matched using propensity score matching (PSM). Cumulative incidence (CI) of local recurrence (LR), locoregional recurrence (LRR), overall survival (OS) and disease-free survival (DFS) were evaluated with a log-rank test. Acute skin toxicity and late radiation pneumonia was analyzed using chi-square test. RESULTS: A total of 529 patients were included in this study, among whom 254 (48%) patients received bolus application. The median follow-up time was 60 months. After matching, 175 well-paired patients were selected. The adjusted 5-year outcomes (95% confidence interval) in patients treated with and without bolus were, respectively: CI of LR 2.42% (0.04-4.74) versus 2.38% (0.05-4.65), CI of LRR 2.42% (0.04-4.74) versus 3.59% (0.73-6.37), DFS 88.12% (83.35-93.18) versus 84.69% (79.42-90.30), OS 94.21% (90.79-97.76) versus 95.86% (92.91-98.91). No correlation between bolus application and skin toxicity (P = 0.555) and late pneumonia (P = 0.333) was observed. CONCLUSIONS: The study revealed a low recurrence rate using IMRT technology. The daily used 5 mm chest wall bolus was not associated with improved clinical outcomes.
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Neoplasias da Mama , Mastectomia , Radioterapia de Intensidade Modulada , Humanos , Feminino , Radioterapia de Intensidade Modulada/efeitos adversos , Radioterapia de Intensidade Modulada/métodos , Neoplasias da Mama/radioterapia , Neoplasias da Mama/cirurgia , Neoplasias da Mama/patologia , Estudos Retrospectivos , Pessoa de Meia-Idade , China/epidemiologia , Adulto , Recidiva Local de Neoplasia/patologia , IdosoRESUMO
BACKGROUND: Telomerase, by safeguarding damaged telomeres and bolstering DNA damage repair, has the capacity to heighten the radioresistance of tumour cells. Thus, in turn, can compromise the efficacy of radiotherapy (RT) and radioimmunotherapy. Our previous studies have revealed that the highly selective telomerase inhibitor, BIBR1532, possesses the potential to enhance the radiosensitivity of Non-small cell lung cancer (NSCLC). In this study, we delve further into the impact of BIBR1532 on the immune activation induced by RT and elucidate the underlying mechanisms. METHODS: Biological information analyses, immunofluorescence assays, western blot assays, flow cytometry analysis were conducted to elucidate the functions of the combination of BIBR1532 with radiotherapy in NSCLC. Intracellular levels of lipid peroxides, glutathione, malondialdehyde, and Fe2+ were measured as indicators of ferroptosis status. Both in vitro and in vivo studies were conducted to examine the antitumor effects. RESULTS: Our findings indicate that the confluence of BIBR1532 with RT significantly augments the activation of the cGAS-STING pathway in both in vivo and in vitro settings, thereby fostering an effective anti-tumoral immune response. The effects can be ascribed to two key processes. Firstly, ionizing radiation, in precipitating DNA double-strand breaks (DSBs), prompts the release of tumour-derived double-stranded DNA (dsDNA) into the cytoplasm. Subsequently, BIBR1532 amplifies the activation of antigen-presenting cells by dsDNA post-RT and instigates the cGAS-STING pathway. Secondly, BIBR1532 enhances the ferroptosis response in NSCLC following RT, thereby promoting unrestrained lipid peroxidation and elevated levels of reactive oxygen species (ROS) within tumour cells. This ultimately leads to mitochondrial stress and the release of endogenous mitochondrial DNA (mtDNA) into the cytoplasm, thus facilitating the activation of the STING pathway and the induction of a type I interferon (IFN)-linked adaptive immune response. CONCLUSION: This study underscores the potential of BIBR1532 as an efficacious and safe radiosensitizer and radioimmunotherapy synergist, providing robust preclinical research evidence for the treatment of NSCLC.
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Carcinoma Pulmonar de Células não Pequenas , Ferroptose , Neoplasias Pulmonares , Proteínas de Membrana , Nucleotidiltransferases , Transdução de Sinais , Carcinoma Pulmonar de Células não Pequenas/radioterapia , Carcinoma Pulmonar de Células não Pequenas/patologia , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/imunologia , Humanos , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/radioterapia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/imunologia , Proteínas de Membrana/metabolismo , Transdução de Sinais/efeitos da radiação , Nucleotidiltransferases/metabolismo , Linhagem Celular Tumoral , Animais , Imunidade/efeitos da radiação , Camundongos Nus , CamundongosRESUMO
The aim was to explore the anti-tumour mechanism of linarin (LIN) based on network pharmacology and molecular docking. PharmMapper database and GeneCards database were used to screen anti-tumour related targets of LIN. Enrichment analysis of GO and KEGG was conducted to predict the key targets and pathways. At last, LIN was docked with the key targets. ESR1, ESR2, EGFR, AR, TGFBR2, F2, MAPK10, MAPK14, CDK2 and HSP90AA1 were identified as the key targets. The key pathways included pathways in cancer, prostate cancer, pancreatic cancer and breast cancer. KEGG pathway maps indicated that the anti-tumour effect of LIN may be mainly achieved by intervening related targets in the following pathways: AR-HSP/AR-AR/PSA/proliferation and evading apoptosis;F2/GPCR/ /ROCK/tissue invasion and metastasis;F2/GPCR/ /Raf/MAPK signalling pathway/proliferation and sustained angiogenesis; EGFR/Grb2/ /Raf/MAPK signalling pathway/proliferation and sustained angiogenesis; ER/Oestrogen signalling pathway/proliferation;TGFBR2/Smad2/3/TGF-ß signalling pathway/insensitivity to anti-growth signals; oxidative stress/KEAP1/NRF2/ /proliferation and evading apoptosis. LIN had strong binding activity with ESR2, EGFR, AR, CDK2 and HSP90AA1.
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The aim of this study was to investigate the expression and function of the transient receptor potential vanilloid 1 (TRPV1) in glioma. We found that the expression of TRPV1 mRNA and protein were upregulated in glioma compared with normal brain by qPCR and western blot analysis. In order to investigate the function of TRPV1 in glioma, short hairpin RNA (shRNA) and the inhibitor of TRPV1 were used. In vitro, the activation of TRPV1 induced cell apoptosis with decreased migration capability and inhibited proliferation, which was abolished upon TRPV1 pharmacological inhibition and silencing. Mechanistically, TRPV1 modulated glioma proliferation through the protein kinase B (Akt) signaling pathway. More importantly, in immunodeficient (NOD-SCID) mouse xenograft models, tumor size was significantly increased when TRPV1 expression was disrupted by a shRNA knockdown approach in vivo. Altogether, our findings indicate that TRPV1 negatively controls glioma cell proliferation in an Akt-dependent manner, which suggests that targeting TRPV1 may be a potential therapeutic strategy for glioma.
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Neoplasias Encefálicas , Proliferação de Células , Glioma , Canais de Cátion TRPV , Animais , Humanos , Camundongos , Apoptose/fisiologia , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/genética , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Glioma/genética , Glioma/metabolismo , Glioma/patologia , Camundongos Endogâmicos NOD , Camundongos SCID , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Interferente Pequeno/farmacologia , Transdução de Sinais/fisiologia , Canais de Cátion TRPV/metabolismo , Canais de Cátion TRPV/genéticaRESUMO
Pea protein isolate (PPI)-based active films were prepared by incorporating 0.5 %, 1.0 %, or 2.0 % of oregano essential oil (OEO), either in the form of micro-emulsion (MOEO) or nano-emulsion (NOEO). The particle size and polydispersity index of OEO droplets were 2755.00 nm and 0.63 for MOEO, and 256.30 nm and 0.20 for NOEO. The surface and cross-sectional SEM results revealed the presence of holes and internal pores within the film upon the addition of OEO. The molecular interaction between PPI and OEO was confirmed by FTIR. The addition of OEO significantly increased film thickness, decreased water contact angle, and imparted a more yellow color. At a low concentration (0.5 %), the addition of OEO significantly improved the water vapor barrier and mechanical properties of the film. However, at higher concentrations, these film properties were significantly weakened. Additionally, the film antimicrobial properties were assessed after OEO addition. In vitro inhibition zone results indicated that a 2.0 % addition of OEO significantly suppressed the growth of three Salmonella strains [Salmonella Typhimurium (ATCC14028), Salmonella Infantis 94-1, and Salmonella Enteritidis PT-30]. Application of pea protein-based film with 2.0 % OEO on chicken breast demonstrated significant reduction in microbial count. Our results further showed that reducing the particle size of OEO from micrometer-scale to nanometer-scale in the PPI film matrix did not significantly alter film properties or antimicrobial activities. The study demonstrated that the antibacterial film based on pea protein and OEO is an innovative food packing material for prohibiting bacteria growth on poultry products.
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BACKGROUND: We aimed to investigate the effects of PinX1 on non-small cell lung cancer(NSCLC) radiosensitivity and radiotherapy-associated tumor immune microenvironment and its mechanisms. METHODS: The effect of PinX1 silencing on radiosensitivity in NSCLC was assessed by colony formation and CCK8 assay, immunofluorescence detection of γ- H2AX and micronucleus assay. Western blot was used to assess the effect of PinX1 silencing on DNA damage repair pathway and cGAS-STING pathway. The nude mouse and Lewis lung cancer mouse model were used to assess the combined efficacy of PinX1 silencing and radiotherapy in vivo. Changes in the tumor immune microenvironment were assessed by flow cytometry for different treatment modalities in the Lewis luuse model. The interaction protein RBM10 was screened by immunoprecipitation-mass spectrometry. RESULTS: Silencing PinX1 enhanced radiosensitivity and activation of the cGAS-STING pathway while attenuating the DNA damage repair pathway. Silencing PinX1 further increases radiotherapy-stimulated CD8+ T cell infiltration and activation, enhances tumor control and improves survival in vivo; Moreover, PinX1 downregulation improves the anti-tumor efficacy of radioimmunotherapy, increases radioimmune-stimulated CD8+ T cell infiltration, and reprograms M2-type macrophages into M1-type macrophages in tumor tissues. The interaction of PinX1 and RBM10 may promote telomere maintenance by assisting telomerase localization to telomeres, thereby inhibiting the immunostimulatory effects of IR. CONCLUSIONS: In NSCLC, silencing PinX1 significantly contributed to the radiosensitivity and promoted the efficacy of radioimmunotherapy. Mechanistically, PinX1 may regulate the transport of telomerase to telomeres through interacting with RBM10, which promotes telomere maintenance and DNA stabilization. Our findings reveal that PinX1 is a potential target to enhance the efficacy of radioimmunotherapy in NSCLC patients.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Telomerase , Animais , Camundongos , Humanos , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/radioterapia , Proteínas Supressoras de Tumor/genética , Proteínas de Ciclo Celular/metabolismo , Telomerase/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/radioterapia , Linhagem Celular Tumoral , Tolerância a Radiação , Microambiente Tumoral , Proteínas de Ligação a RNARESUMO
Biofilms formed by pathogenic bacteria present a persistent risk to human health. While the eradication of matured biofilms remains a formidable challenge, delaying or preventing their formation, which is coordinately regulated by quorum sensing (QS), presents a simpler and more advantageous strategy. Quercetin, a naturally occurring compound with anti-QS properties, has the potential to act as an antibiofilm agent. However, it is plagued by certain inherent drawbacks, including poor water solubility and limited bioavailability. Furthermore, solely blocking QS is not enough to prevent biofilm formation because it lacks bactericidal properties. To address these difficulties, we fabricated bi-functional nanoparticles through the co-assembly of quercetin and copper ions in a facile manner. The resulting quercetin/copper nanoparticles (QC NPs) demonstrated minimal cytotoxicity and hemolysis in vitro. In response to the low pH of microenvironments that were populated by bacterial colonies, the QC NPs underwent disassembly to release copper ions and quercetin. The former exterminated bacteria by disrupting the integrity of the cell membrane, while the latter disrupted the processes involved in QS that are responsible for the biofilm by downregulating the expression of specific genes, effectively preventing the formation of biofilms by both Gram-negative Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus. In addition, the QC NPs were integrated into a bacterial cellulose membrane. The composite membrane proved to be highly effective at inhibiting biofilm formation in vitro and demonstrated the ability to reduce inflammatory responses and accelerate the healing of bacteria-infected wounds in vivo. Overall, the bi-functional QC NPs hold great potential for use in addressing the challenges associated with the management of bacterial biofilms.
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Nanopartículas , Percepção de Quorum , Humanos , Quercetina/farmacologia , Cobre/farmacologia , Biofilmes , Antibacterianos/farmacologia , Antibacterianos/química , Bactérias , Íons/farmacologia , Pseudomonas aeruginosaRESUMO
Oxaliplatin (OXA)-based chemotherapy is one of the first-line treatments for advanced gastric cancer. However, the potential risk for chemotherapy-induced hepatic injury can hinder its effectiveness. Polyene phosphatidylcholine (PPC) is often used as a hepatoprotective agent to counter OXA-induced hepatic injury; however, its impact on the antitumour effectiveness of OXA remains uncertain. Our retrospective study examined 98 patients with stage IV gastric cancer to assess the impact of PPC on progression-free survival (PFS) and disease control rate (DCR). Furthermore, in vitro and in vivo assays were conducted to elucidate the combined biological effects of OXA and PPC (OXA+PPC) on gastric cancer. RNA sequencing, luciferase reporter assays, live/dead cell assays, immunofluorescence, and western blotting were used to identify the activated signalling pathways and downstream factors post OXA+PPC treatment. The findings indicated that PPC served as an independent prognostic factor, correlating with prolonged PFS and improved DCR in patients with gastric cancer. The combination of OXA and PPC significantly inhibited tumour cell growth both in vitro and in vivo. RNA sequencing revealed that OXA+PPC treatment amplified reactive oxygen species and ferroptosis signalling pathways. Mechanistically, OXA+PPC upregulated the expression of haem oxygenase-1 by promoting the nuclear migration of nuclear factor erythroid 2-related factor (Nrf2), thereby enhancing its transcriptional activity. Drug-molecule docking analysis demonstrated that PPC competitively bound to the peptide structural domains of both Nrf2 and Kelch-like ECH-associated protein 1 (KEAP1), accounting for the increased translocation of Nrf2. In conclusion, our study reveals the synergistic antitumour potential of PPC and OXA while protecting patients against hepatic injury. This suggests a promising combined treatment approach for patients with advanced gastric cancer.
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INTRODUCTION: Abnormal alternative splicing (AS) contributes to aggressive intrahepatic invasion and metastatic spread, leading to the high lethality of hepatocellular carcinoma (HCC). OBJECTIVES: This study aims to investigate the functional implications of UPF3B-S (a truncated oncogenic splice variant) in HCC metastasis. METHODS: Basescope assay was performed to analyze the expression of UPF3B-S mRNA in tissues and cells. RNA immunoprecipitation, and in vitro and in vivo models were used to explore the role of UPF3B-S and the underlying mechanisms. RESULTS: We show that splicing factor HnRNPR binds to the pre-mRNA of UPF3B via its RRM2 domain to generate an exon 8 exclusion truncated splice variant UPF3B-S. High expression of UPF3B-S is correlated with tumor metastasis and unfavorable overall survival in patients with HCC. The knockdown of UPF3B-S markedly suppresses the invasive and migratory capacities of HCC cells in vitro and in vivo. Mechanistically, UPF3B-S protein targets the 3'-UTR of CDH1 mRNA to enhance the degradation of CDH1 mRNA, which results in the downregulation of E-cadherin and the activation of epithelial-mesenchymal transition. Overexpression of UPF3B-S enhances the dephosphorylation of LATS1 and the nuclear accumulation of YAP1 to trigger the Hippo signaling pathway. CONCLUSION: Our findings suggest that HnRNPR-induced UPF3B-S promotes HCC invasion and metastasis by exhausting CDH1 mRNA and modulating YAP1-Hippo signaling. UPF3B-S could potentially serve as a promising biomarker for the clinical management of invasive HCC.
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Anaplastic thyroid carcinoma (ATC) is an aggressive malignancy threatening patients' life quality. Our previous study has demonstrated that inhibition of long non-coding RNA H19 (lncRNA h19; H19) blocks ATC growth and metastasis. The current study aimed to further explore the potential mechanism of H19 in ATC. Expression of H19, miR-454-3p, and BHLHE40 mRNA was measured using RT-qPCR in tissue samples and cell lines. The dual-luciferase reporter assay and Pearson correlation analysis were used to explore the interaction among H19, miR-454-3p, and BHLHE40. The biological process of proliferation, migration, and invasion was determined using loss- or gain-function CCK-8 and Transwell assays. Western blot assay was used to evaluate the changes in protein levels. H19 was elevated in ATC tissues and cell lines. Based on online prediction database results, miR-454-3p might be a target of H19, and BHLHE40 might be a direct target of miR-454-3p. miR-454-3p expression was decreased in ATC and had a negative interaction with H19. BHLHE40 mRNA expression was increased and has a negative correlation with miR-454-3p and a positive correlation with H19. Downregulation of miR-454-3p and upregulation of BHLHE40 could reverse the decreased cellular activities caused by si-H19. Moreover, the silence of H19 modulates BHLHE40 to affect the PI3K/AKT protein levels and apoptotic-related protein levels. The current study provided a potential detailed mechanism of H19 in ATC, and lncRNA H19-miR-454-3p-BHLHE40 interaction may be a new experimental basis for prognosis and targeted therapy for ATC patients.
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Selenium nanoparticles (Se NPs) have significant anticancer effects, but their poor water solubility and dispersibility limit their further applications in biomedical fields. Biomacromolecules have often been used as dispersants or stabilizers in synthesized Se NPs because they can enhance the dispersibility of Se NPs and reduce their side effects. Our previous studies reported a triple-helix ß-glucan (BFP) from the fruiting bodies of black fungus, which showed a good self-assembly ability in constructing hollow nanotubes for loading metal nanoparticles. Therefore, in the present work, BFP nanotubes were designed as carriers to entrap large amounts of Se NPs in order to enhance their stability and anticancer effects. The results showed that Se NPs were successfully synthesized and loaded inside the BFP nanotubes, and the composite (BFP-Se) exhibited high stability and dispersibility due to the covalent Se-O bonds between the Se NPs and the hydroxyl groups on the BFP nanotubes. Moreover, BFP-Se showed significant effects on the proliferation, apoptosis, and cell cycle of HepG2 cells compared to those exhibited by Se NPs. The mechanism was associated with BFP, which acted as a dispersant or stabilizer, resulting in the enhanced cellular uptake of the Se NPs. BFP also activated the death receptor-mediated and mitochondria-mediated apoptotic pathways in HepG2 cells. These results suggest that BFP-Se has potential applications in biomedical fields, especially for the treatment of human liver cancers.
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Most reported polysaccharides from Poria cocos (PCPs) in traditional Chinese medicine decoctions were water-soluble heteropolysaccharides while the water-insoluble PCPs were scarcely researched due to the poor water-solubility. In this study, a water-insoluble polysaccharide with high yield of 59%, and high purity with a glucan content of 98.8%, was isolated by diluted sodium hydroxide at low temperature and coded as PCPA. The chemical structure of PCPA was identified as a liner ß-glucan with 1, 3-linked glycosidic bond by the fourier infrared spectrum (FT-IR), ion chromatography (ICP), gas chromatography and mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) measurements. Importantly, PCPA was successfully used to construct hydrogels (PCPA-Gs) with good thermal stability, water retention ability and swelling property through simple physical cross-linking, due to the abundance of hydroxyl groups on glucan chains. Moreover, the rheology analysis of PCPA-Gs showed a rapid transition between gel and sol as well as the shear-thinning property. The hydrogel developed in this study holds promise for applications in the food, pharmaceutical, and cosmetic fields.
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Wolfiporia , beta-Glucanas , Wolfiporia/química , Água , Hidrogéis , Espectroscopia de Infravermelho com Transformada de Fourier , Polissacarídeos/químicaRESUMO
Adipogenesis is tightly regulated by various factors, including genes and microRNAs. Excessive fat deposition is the key feature of obesity, which is a low-grade chronic inflammatory disease. Follistatin-like 1 (FSTL1) has been reported to be an important mediator involved in various inflammatory diseases. However, the underlying mechanism of FSTL1 in preadipocyte differentiation and inflammatory response is still unclear. The current study was designed to explore the biological function and potential mechanism of FSTL1 in mouse subcutaneous preadipocyte differentiation. We found that FSTL1 was highly expressed in the early stage of differentiation and subsequently decreased sharply, suggesting that FSTL1 played a possible role in adipogenesis. Meanwhile, the gain- and loss-of-function assays showed that FSTL1 was not only involved in the inflammatory response by inducing the expression of pro-inflammatory factors IL-1ß and CCL2 but also significantly attenuated preadipocyte differentiation, as evidenced by the reduction of lipid accumulation and the levels of adipogenic genes, including PPARγ and FABP4. In addition, the target gene prediction and luciferase reporter assay validated that miR-125a-3p targeted the 3' UTR region of FSTL1. These results demonstrated that miR-125a-3p negatively regulated the expression of FSTL1 at the mRNA and protein levels. Furthermore, overexpressing miR-125a-3p in preadipocytes dramatically accelerated adipogenic differentiation and downregulated the levels of IL-1ß and CCL2, which were in accordance with the knockdown of FSTL1. On the contrary, treatment with miR-125a-3p inhibitors attenuated adipogenesis but induced the expression of inflammatory genes. In summary, this study suggests a positive function of FSTL1 in adipocyte-induced inflammation and negatively regulates preadipocyte differentiation. Further studies demonstrated that miR-125a-3p could reverse the effect by targeting FSTL1, which might provide a better understanding of treating obesity-related inflammatory diseases.
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Adipogenia , MicroRNAs , Animais , Camundongos , Adipócitos/metabolismo , Adipogenia/genética , MicroRNAs/metabolismo , Obesidade/genética , Obesidade/metabolismo , RNA Mensageiro/metabolismoRESUMO
One of the serious threats to global public health is the bacterial biofilm, which results in numerous persistent and recurrent infections. Herein, we proposed a near-infrared (NIR) light-triggered "nano-domino" system with "dispersing and killing" functionality for biofilm eradication. The nanoplatform was fabricated by the self-assembly of chitosan conjugated with L-arginine (L-Arg, a natural nitric oxide (NO) donor) and indocyanine green (ICG, a phototherapy agent). Using an NIR irradiation "trigger", a series of reactive oxygen species (ROS) including singlet oxygen (1O2), hydrogen peroxide (H2O2), and superoxide anions (·O2-), as well as heat were generated from ICG aggregates. Subsequently, 1O2 and H2O2 catalyzed L-Arg to produce NO, which dispersed the biofilm and reacted with ·O2- to form peroxynitrite to kill bacteria with ROS collaboratively. Meanwhile, the generated heat increased the permeability of bacterial membranes, aggravating the damage to biofilm bacteria. The experiments on biofilm eradication demonstrated that this "nano-domino" system was capable to eradicate over 99.99% of biofilms formed by Methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa under 5-min NIR irradiation. Notably, these integrated benefits allowed the system to promote the healing of MRSA biofilm-infected wounds in vivo with negligible toxicity. Overall, this reported NIR-triggered "nano-domino" system holds great promise for addressing the difficulties associated with bacterial biofilm eradication. STATEMENT OF SIGNIFICANCE: Novel agents for biofilm eradication are urgently needed due to the alarming rise in antimicrobial resistance to conventional antibiotics and the critical shortage of new drugs. In this study, we created a nano-domino system that uses near-infrared (NIR) light as a trigger to eradicate mature biofilms. In response to a short-term NIR irradiation, the proposed nanoplatform could generate nitric oxide and peroxynitrite to disperse the biofilm and kill the bacteria inside, respectively, leading to efficient eradication of Methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa biofilms with minimal cytotoxicity. The findings, therefore, indicate that this nanoplatform with enhanced antibiofilm performance might provide a reliable and promising solution to biofilm-related problems.
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Intracellular delivery of bioactive macromolecules and functional materials plays a crucial role in fundamental biological research and clinical applications. Nondestructive and efficient harvesting of engineered cells is also required for some specific applications. In this work, we develop a multifunctional platform based on candle soot modified with copolymer brushes containing temperature-responsive poly(N-isopropylacrylamide) (PNIPAAm) and sugar-responsive phenylboronic acid (PBA) components. This platform possesses a high cell adhesion capacity due to the inherent hierarchical structure of candle soot and the formation of boronate ester bonds between the PBA groups and glycoproteins on the cell membrane. Under the irradiation of a near-infrared laser, the excellent light-to-heat conversion ability of candle soot enables the highly efficient delivery of macromolecules into diverse cells (including hard-to-transfect cells) attached to the surface via a photothermal-poration mechanism. Owing to the temperature-responsive properties of PNIPAAm and the sugar-responsive properties of PBA, the engineered cells could be harvested nondestructively from the platform by a mild treatment using a cold fructose solution. A proof-of-concept experiment demonstrates that fibroblasts attached to the surface could be transfected by a functional plasmid encoding basic fibroblast growth factor and then harvested efficiently and recultured with improved proliferation and migration ability. The whole delivery-harvesting process required less than 1 h, allowing the cells to be engineered without compromising their viability. This platform thus provides a widely applicable method for both the intracellular delivery of diverse macromolecules efficiently as well as harvesting engineered cells simply and safely, holding great potential for biomedical applications.
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Ácidos Borônicos , Fuligem , AçúcaresRESUMO
The aim of this study is to develop an antioxidant film based on pea protein isolate (PPI) without sacrificing the packaging properties. To achieve this, α-tocopherol was incorporated to impart antioxidant activity to the film. We investigated the effects on film properties resulting from the addition of α-tocopherol in a nanoemulsion form and pH-shifting treatment of PPI. The results revealed that direct addition of α-tocopherol into un-treated PPI film disrupted film structure and formed a discontinuous film with rough surface, and thereby significantly decreasing the tensile strength and elongation at break. However, pH-shifting treatment in combination with the α-tocopherol nanoemulsion, formed a smooth and compact film, which greatly improved the mechanical properties. It also significantly changed the color and opacity of PPI film, but had little effects on film solubility, moisture content, and water vapor permeability. After the addition of α-tocopherol, the DPPH scavenging ability of PPI film was greatly improved and the release of α-tocopherol was mainly within the first 6 h. Additionally, pH-shifting and nanoemulsion did not affect the film's antioxidant activity nor the release rate. In conclusion, pH-shifting combined with nanoemulsion is an effective method to incorporate hydrophobic compounds such as α-tocopherol into protein-based edible films without negative impacts on film mechanical properties.