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1.
Anal Chim Acta ; 1315: 342760, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38879206

RESUMO

Mycotoxins are commonly found in food materials and severely threaten human health. Antibodies play a key role as a part of immunological techniques in detecting mycotoxins. Therefore, highly specific antibodies and detection techniques against mycotoxins need to be developed for advancements in medical research. In this study, we presented a novel strategy for quickly screening highly specific antigen-binding fragment (Fab) antibodies based on yeast surface display (YSD) and detecting small-molecule compounds based on a YSD biosensor. We constructed a yeast surface display Deoxynivalenol (DON)-Fab library with 105 cfu/mL with a galactose-inducible bidirectional promoter. By conducting efficient magnetic-activated cell sorting and fluorescence-activated cell sorting (MACS/FACS), four kinds of DON-selective yeasts were screened. As Fab@YSD C4# showed high sensitivity, we used it to build a one-pot Fab@YSD chemiluminescence biosensor with DON-BSA@Biotin and Streptavidin-alkaline phosphatase (SA-ALP). This method showed a low operational threshold (LOD = 0.166 pg/mL) and a high population range (linear range = 0.001-132.111 ng/mL) within 40 min, which facilitated the detection of DON with high specificity and better recovery in real samples (wheat, corn, flour, and cornmeal). Our results suggested that the Fab@YSD chemiluminescence biosensor is an inexpensive, reproducible, user-friendly, and sensitive method for detecting DON and may be used to quickly detect other small-molecule contaminants in food items.


Assuntos
Técnicas Biossensoriais , Tricotecenos , Tricotecenos/análise , Técnicas Biossensoriais/métodos , Saccharomyces cerevisiae , Contaminação de Alimentos/análise , Fragmentos Fab das Imunoglobulinas/química , Fragmentos Fab das Imunoglobulinas/imunologia , Limite de Detecção , Triticum/química , Triticum/microbiologia , Zea mays/química , Zea mays/microbiologia , Farinha/análise
2.
FASEB J ; 38(7): e23565, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38558188

RESUMO

Circadian rhythms in metabolically active tissues are crucial for maintaining physical health. Circadian disturbance (CD) can cause various health issues, such as metabolic abnormalities and immune and cognitive dysfunctions. However, studies on the role of CD in immune cell development and differentiation, as well as the rhythmic expression of the core clock genes and their altered expression under CD, remain unclear. Therefore, we exposed C57bl/6j mice to repeated reversed light-dark cycles for 90 days to research the effects of CD on bone marrow (BM) hematopoietic function. We also researched the effects of CD on endogenous circadian rhythms, temporally dependent expression in peripheral blood and myeloid leukocytes, environmental homeostasis within BM, and circadian oscillations of hematopoietic-extrinsic cues. Our results confirmed that when the light and dark cycles around mice were frequently reversed, the circadian rhythmic expression of the two main circadian rhythm markers, the hypothalamic clock gene, and serum melatonin, was disturbed, indicating that the body was in a state of endogenous CD. Furthermore, CD altered the temporally dependent expression of peripheral blood and BM leukocytes and destroyed environmental homeostasis within the BM as well as circadian oscillations of hematopoietic-extrinsic cues, which may negatively affect BM hematopoiesis in mice. Collectively, these results demonstrate that circadian rhythms are vital for maintaining health and suggest that the association between CD and hematopoietic dysfunction warrants further investigation.


Assuntos
Medula Óssea , Relógios Circadianos , Camundongos , Animais , Medula Óssea/metabolismo , Fotoperíodo , Ritmo Circadiano/fisiologia , Células-Tronco Hematopoéticas/metabolismo , Camundongos Endogâmicos C57BL , Relógios Circadianos/genética
3.
ACS Appl Mater Interfaces ; 15(5): 6338-6353, 2023 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-36701257

RESUMO

Bacteria-infected skin wounds caused by external injuries remain a serious challenge to the whole society. Wound healing dressings, with excellent antibacterial activities and potent regeneration capability, are increasingly needed clinically. Here, we reported a novel functional microneedle (MN) array comprising methacrylated hyaluronic acid (MeHA) embedded with pH-responsive functionalized zeolitic imidazolate framework-8 (ZIF-8) nanoparticles to treat bacteria-infected cutaneous wounds. Antibacterial activity was introduced into Zn-ZIF-8 to achieve sterilization through releasing Zn ions, as well as increased angiogenesis by dimethyloxalylglycine (DMOG) molecules that were distributed within its framework. Furthermore, biodegradable MeHA was chosen as a substrate material carrier to fabricate DMOG@ZIF-8 MN arrays. By such design, DMOG@ZIF-8 MN arrays would not only exhibit excellent antibacterial activity against pathogenic bacteria but also enhance angiogenesis within wound bed by upregulating the expression of HIF-1α, leading to a significant therapeutic efficiency on bacteria-infected cutaneous wound healing. Based on these results, we conclude that this new treatment strategy can provide a promising alternative for accelerating infected wound healing via effective antibacterial activity and ameliorative angiogenesis.


Assuntos
Sistemas de Liberação de Medicamentos , Nanopartículas , Zeolitas , Antibacterianos/administração & dosagem , Antibacterianos/farmacologia , Bactérias , Nanopartículas/química , Zeolitas/química , Cicatrização
4.
Front Pharmacol ; 14: 1302059, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38259290

RESUMO

Objective: This study aimed to investigate the molecular mechanism of triptolide in the treatment of oral squamous cell carcinoma (OSCC) via network pharmacology and experimental validation. Methods: The network pharmacological method was used to predict the key targets, detect the signal pathways for the treatment of OSCC, and screen the critical components and targets for molecular docking. Predicted targets were validated in cellular and xenograft mouse model. Results: In this study, we predicted action on 17 relevant targets of OSCC by network pharmacology. PPI network demonstrated that Jun, MAPK8, TP53, STAT3, VEGFA, IL2, CXCR4, PTGS2, IL4 might be the critical targets of triptolide in the treatment of OSCC. These potential targets are mainly closely related to JAK-STAT and MAPK signaling pathways. The analysis of molecular docking showed that triptolide has high affinity with Jun, MAPK8 and TP53. Triptolide can suppress the growth of OSCC cells and xenograft mice tumor, and downregulate the expression of Jun, MAPK8, TP53, STAT3, VEGFA, IL2, CXCR4, PTGS2 to achieve the therapeutic effect of OSCC. Conclusion: Through network pharmacological methods and experimental studies, we predicted and validated the potential targets and related pathways of triptolide for OSCC treatment. The results suggest that triptolide can inhibit the growth of OSCC via several key targets.

5.
Proteomics ; 20(14): e1900423, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32468662

RESUMO

High-altitude polycythemia (HAPC) is a common plateau chronic disease in which red blood cells are compensatory hyperproliferative due to high altitude hypoxic environment. HAPC severely affects the physical and mental health of populations on the plateau. However, the pathogenesis and treatment of HAPC has been rarely investigated. Here, the hypoxia-induced HAPC model of rat is established, in which hemoglobin concentration significantly increases and platelets clearly decrease. The effect of resveratrol upon hypoxia enables HAPC remission and makes hemoglobin and platelet tend to a normal level. Furthermore, quantitative proteomics is applied to investigate the plasma proteome variation and the underlying molecular regulation during HAPC occurrence and treatment with resveratrol. Hypoxia promotes erythrocyte developing and differentiating and disrupts cytoskeleton organization. Notably, the resveratrol administration reverses the proteome change pattern due to hypoxia and contributes to plateau adaption. Quantitative verification of differentially expressed proteins confirms the roles of resveratrol in HAPC. Resveratrol is expected to be useful for HAPC treatment.


Assuntos
Doença da Altitude/complicações , Altitude , Hipóxia/fisiopatologia , Policitemia/tratamento farmacológico , Proteoma/metabolismo , Resveratrol/farmacologia , Transcriptoma/efeitos dos fármacos , Adaptação Fisiológica , Animais , Antioxidantes/farmacologia , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Masculino , Policitemia/etiologia , Policitemia/metabolismo , Policitemia/patologia , Proteoma/análise , Proteoma/efeitos dos fármacos , Ratos , Ratos Wistar
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