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1.
Mikrochim Acta ; 191(5): 289, 2024 04 29.
Artigo em Inglês | MEDLINE | ID: mdl-38683210

RESUMO

As a common chlorinated nicotinic pesticide with high insecticidal activity, acetamiprid has been widely used for pest control. However, the irrational use of acetamiprid will pollute the environment and thus affect human health. Therefore, it is crucial to develop a simple, highly sensitive, and rapid method for acetamiprid residue detection. In this study, the capture probe (Fe3O4@Pt-Aptamer) was connected with the signal probe (Au@DTNB@Ag CS-cDNA) to form an assembly with multiple SERS-enhanced effects. Combined with magnetic separation technology, a SERS sensor with high sensitivity and stability was constructed to detect acetamiprid residue. Based on the optimal conditions, the SERS intensity measured at 1333 cm-1 is in relation to the concentration of acetamiprid in the range 2.25 × 10-9-2.25 × 10-5 M, and the calculated limit of detection (LOD) was 2.87 × 10-10 M. There was no cross-reactivity with thiacloprid, clothianidin, nitenpyram, imidacloprid, and chlorpyrifos, indicating that this method has good sensitivity and specificity. Finally, the method was applied to the detection of acetamiprid in cucumber samples, and the average recoveries were 94.19-103.58%, with RSD < 2.32%. The sensor can be used to analyse real samples with fast detection speed, high sensitivity, and high selectivity.


Assuntos
Aptâmeros de Nucleotídeos , Ouro , Limite de Detecção , Nanopartículas Metálicas , Neonicotinoides , Prata , Análise Espectral Raman , Neonicotinoides/análise , Aptâmeros de Nucleotídeos/química , Ouro/química , Prata/química , Nanopartículas Metálicas/química , Análise Espectral Raman/métodos , Platina/química , Inseticidas/análise , Cucumis sativus/química
2.
Eur Arch Psychiatry Clin Neurosci ; 273(6): 1267-1277, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36567366

RESUMO

The lack of objective diagnostic methods for mental disorders challenges the reliability of diagnosis. The study aimed to develop an easily accessible and useable objective method for diagnosing major depressive disorder (MDD), schizophrenia (SZ), bipolar disorder (BPD), and panic disorder (PD) using serum multi-protein. Serum levels of brain-derived neurotrophic factor (BDNF), VGF (non-acronymic), bicaudal C homolog 1 (BICC1), C-reactive protein (CRP), and cortisol, which are generally recognized to be involved in different pathogenesis of various mental disorders, were measured in patients with MDD (n = 50), SZ (n = 50), BPD (n = 55), and PD along with 50 healthy controls (HC). Linear discriminant analysis (LDA) was employed to construct a multi-classification model to classify these mental disorders. Both leave-one-out cross-validation (LOOCV) and fivefold cross-validation were applied to validate the accuracy and stability of the LDA model. All five serum proteins were included in the LDA model, and it was found to display a high overall accuracy of 96.9% when classifying MDD, SZ, BPD, PD, and HC groups. Multi-classification accuracy of the LDA model for LOOCV and fivefold cross-validation (within-study replication) reached 96.9 and 96.5%, respectively, demonstrating the feasibility of the blood-based multi-protein LDA model for classifying common mental disorders in a mixed cohort. The results suggest that combining multiple proteins associated with different pathogeneses of mental disorders using LDA may be a novel and relatively objective method for classifying mental disorders. Clinicians should consider combining multiple serum proteins to diagnose mental disorders objectively.


Assuntos
Transtorno Depressivo Maior , Transtornos Mentais , Humanos , Transtorno Depressivo Maior/diagnóstico , Reprodutibilidade dos Testes , Transtornos Mentais/diagnóstico , Proteínas Sanguíneas , Aprendizado de Máquina
3.
Mikrochim Acta ; 191(1): 29, 2023 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-38095724

RESUMO

A novel nucleic acid aptamer nanoprobes-mediated hairpin allosteric and aptamer-assisted CRISPR system for detection of Streptococcus pneumoniae and Staphylococcus aureus is presented. In this fluorescence assay system, utilizing the hairpin allosteric effect caused by the aptamer binding to the target bacteria, the detection of S. pneumoniae is first achieved through changes in fluorescence due to FRET. Subsequently, a Cas12a protein mixture is added to detect S. aureus. The amplified output signal is triggered by two methods to ensure the sensitivity of the method: the synergistic FRET effect is achieved by the assembly of multi-aptamer through the conjugation of streptavidin-biotin, and the trans-cleavage function of CRISPR/Cas 12a. Under the optimized conditions, the proposed hairpin allosteric aptasensor could achieve high sensitivity (a detection limit of 135 cfu/mL) and broad-concentration quantification (dynamic range of 103-107 cfu/mL) of S. pneumoniae. The aptamer-assisted CRISPR system for S. aureus detection showed good linearity (R2 = 0.996) in the concentration range 102-108 cfu/mL, with a detection limit of 39 cfu/mL. No cross-reactivity with other foodborne pathogenic bacteria was observed in both systems. Taking only 55 min, this method of multiple pathogen detection proved to be promising.


Assuntos
Aptâmeros de Nucleotídeos , Staphylococcus aureus Resistente à Meticilina , Staphylococcus aureus/genética , Aptâmeros de Nucleotídeos/genética , Streptococcus pneumoniae/genética , Bactérias
4.
Hum Brain Mapp ; 42(18): 6070-6086, 2021 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-34597450

RESUMO

The aim of this study is to investigate the trajectory of medial longitudinal fasciculus (MLF) and explore its anatomical relationship with the oculomotor nerve using tractography technique. The MLF and oculomotor nerve were reconstructed at the same time with preset three region of interests (ROIs): one set at the area of rostral midbrain, one placed on the MLF area at the upper pons, and one placed at the cisternal part of the oculomotor nerve. This mapping protocol was tested in an HCP-1065 template, 35 health subjects from Massachusetts General Hospital (MGH), 20 healthy adults and 6 brainstem cavernous malformation (BCM) patients with generalized q-sampling imaging (GQI)-based tractography. Finally, the 200 µm brainstem template from Center for In Vivo Microscopy, Duke University (Duke CIVM), was used to validate the trajectory of reconstructed MLF. The MLF and oculomotor nerve were reconstructed in the HCP-1065 template, 35 MGH health subjects, 20 healthy adults and 6 BCM patients. The MLF was in conjunction with the ipsilateral mesencephalic part of the oculomotor nerve. The displacement of MLF was identified in all BCM patients. Decreased QA, RDI and FA were found in the MLF of lesion side, indicating axonal loss and/or edema of displaced MLF. The reconstructed MLF in Duke CIVM brainstem 200 µm template corresponded well with histological anatomy. The MLF and oculomotor nerve were visualized accurately with our protocol using GQI-based fiber tracking. This GQI-based tractography is an important tool in the reconstruction and evaluation of MLF.


Assuntos
Tronco Encefálico/patologia , Imagem de Tensor de Difusão/métodos , Hemangioma Cavernoso do Sistema Nervoso Central/patologia , Nervo Oculomotor/anatomia & histologia , Substância Branca/anatomia & histologia , Adulto , Tronco Encefálico/diagnóstico por imagem , Feminino , Hemangioma Cavernoso do Sistema Nervoso Central/diagnóstico por imagem , Humanos , Masculino , Pessoa de Meia-Idade , Vias Neurais/anatomia & histologia , Vias Neurais/diagnóstico por imagem , Nervo Oculomotor/diagnóstico por imagem , Nervo Oculomotor/patologia , Substância Branca/diagnóstico por imagem , Substância Branca/patologia , Adulto Jovem
5.
Am J Respir Cell Mol Biol ; 61(3): 380-391, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30908929

RESUMO

Silicosis is a progressive fibrotic disease of lung tissue caused by long-term inhalation of SiO2. However, relatively few studies of the direct effects of SiO2 on lung fibroblasts have been performed. PPP1R13B is a major member of the apoptosis-stimulating proteins of the p53 family, but its role in pulmonary fibrosis is unclear. To elucidate the role of PPP1R13B in the pathological process of silicosis, we explored the molecular mechanisms related to PPP1R13B and the functional effects of proliferation and migration of fibroblasts. Through lentivirus transfection, Western blotting, and fluorescent in situ hybridization experiments, we found that SiO2 downregulated circRNA-012091 (circ-012091) expression in lung fibroblasts and induced upregulation of downstream PPP1R13B. Transfection of L929 cells with PPP1R13B CRISPR NIC plasmid inhibited the upregulation of endoplasmic reticulum stress (ERS) and autophagy-related protein expression in lung fibroblasts treated with SiO2, and induced decreases in cell proliferation, migration, and viability. Transfection of L929 cells with the PPP1R13B CRISPR ACT plasmid induced increases in cell proliferation, migration, and viability. In addition, the ERS inhibitor salubrinal and the autophagy inhibitor 3-methyladenine inhibited the increased migration of L929 cells transfected with the PPP1R13B CRISPR ACT plasmid. These results suggest that PPP1R13B regulated by circ-012091 promotes the proliferation and migration of lung fibroblasts through ERS and autophagy, and plays a crucial role in the development of pulmonary fibrosis in silicosis.


Assuntos
Autofagia/efeitos dos fármacos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Dióxido de Silício/farmacologia , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/metabolismo , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Hibridização in Situ Fluorescente/métodos , Pulmão/patologia , Fibrose Pulmonar/tratamento farmacológico , Fibrose Pulmonar/patologia , RNA Circular/metabolismo , Transdução de Sinais/efeitos dos fármacos
6.
Cell Physiol Biochem ; 51(3): 1389-1398, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30481761

RESUMO

BACKGROUND/AIMS: Hepatic stellate cells (HSCs) are the primary cell type responsible for liver fibrosis. Our study proved that thymosin beta 4 (Tß4) has anti-fibrogenic effects in HSCs through PI3K/AKT pathway. However, the underlying mechanisms are not fully elucidated. Circular RNAs (circRNAs) play important roles in fine-tuning gene expression and are often deregulated in cancers. However, the expression profile and clinical significance of in liver fibrosis is still unknown. Therefore, we hypothesize that Tß4 influences circRNAs in liver fibrosis. METHODS: Circular RNA microarray was conducted to identify Tß4-related circRNAs. Pathway analysis and miRNA response elements analysis was conducted to predict the potential roles of differentially expressed circRNAs in liver fibrosis. CCK8 assays and flow cytometric assays were conducted to clarify the role of circRNA in liver fibrosis. Bioinformatics analysis and in vitro experiments were conducted to clarify the mechanism of circRNA-mediated gene regulation in liver fibrosis. RESULTS: A total of 644 differentially expressed circRNAs were identified between the Tß4-depleted LX-2 cells and the control LX2 cells. The expression of circRNA-0067835 was significantly increased in the Tß4-depleted LX-2 cells compared with control. Knockdown of circRNA-0067835 observably decreased LX-2 cell proliferation by causing G1 arrest and promoting apoptosis. Bioinformatics online programs predicted that circRNA-0067835 acted as miR-155 sponge to regulate FOXO3a expression, which was validated using luciferase reporter assay. CONCLUSION: Our experiments showed that circRNA-0067835 regulated liver fibrosis progression by acting as a sponge of miR-155 to promote FOXO3a expression, indicating that circRNA-0067835 may serve as a potential therapeutic target for patients with liver fibrosis.


Assuntos
Proteína Forkhead Box O3/genética , Regulação da Expressão Gênica , Células Estreladas do Fígado/metabolismo , MicroRNAs/genética , RNA/genética , Transdução de Sinais , Timosina/genética , Animais , Linhagem Celular , Células Cultivadas , Células Estreladas do Fígado/patologia , Humanos , Cirrose Hepática/genética , Cirrose Hepática/patologia , Masculino , Camundongos Endogâmicos C57BL , RNA Circular , Transcriptoma
7.
J Magn Reson Imaging ; 42(4): 1134-43, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26218829

RESUMO

PURPOSE: To investigate the feasibility of T2W-SPACE technique in early detection of WD, the signal evolutions of degenerated corticospinal tract (CST) on T2W-SPACE, and their underlying pathological changes. MATERIALS AND METHODS: The WD model of the CST was established in 23 cats through excision of cortical origins of the tract. Eight cats were scanned with the T2W-SPACE technique at 8 sequential time points, i.e. 0 (before modeling), 2, 4, 6, 8, 10, 20 and 30 days after modeling, and then they were pathologically examined. The remaining 15 cats (3 per group) also underwent pathological examination at 2, 4, 6, 10 and 20 days after modeling, respectively. The ratios of T2 signal intensity (rT2s) between the affected and unaffected sides of CST were analyzed. RESULTS: During the first 4 days, SPACE could not detect any significant changes of the affected CST, although axonal degeneration was pathologically observed at the second day. From 6 to 10 days, the rT2s decreased monotonously, which is corresponded to histological findings of myelin degeneration and phagocyte proliferation. From 10 to 20 days, rT2s kept relatively stable at a low level and started to recover after that; the pathological changes of this period was characterized by marked phagocytizing activities. CONCLUSION: SPACE technique can detect Wallerian degeneration at an early stage, and the signal evolution is consistent with the pathological processes.


Assuntos
Imagem de Tensor de Difusão/métodos , Interpretação de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Tratos Piramidais/patologia , Doenças da Medula Espinal/patologia , Degeneração Walleriana/patologia , Animais , Gatos , Progressão da Doença , Estudos de Viabilidade , Feminino , Aumento da Imagem/métodos , Masculino , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
8.
J Colloid Interface Sci ; 661: 374-388, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38306747

RESUMO

Conventional antibiotic treatment struggles to eliminate biofilms in wounds due to the formation compact barrier. Herein, we fabricate magnetic pandanus fruit-like nanorobots (NRs) that function as drug carriers while exhibit excellent maneuverability for enhanced antibacterial tasks. Specifically, zeolitic imidazolate framework-8 (ZIF-8) is self-assembled on the surface of Fe3O4 nanoparticles, loaded with a small quantity of ciprofloxacin, and covered with a layer of polydopamine (PDA). Energized by external magnetic fields, the NRs (F@Z/C/P) are steered in defined direction to penetrate the infection tissues, and effectively arrive targeted areas for pH stimulated drug release and near-infrared triggered phototherapy, contributing to an antibacterial rate of >99.9 %. The Zn2+ in ZIF-8 and the catechol group in PDA form catechol-ZIF-8-drug structures, which effectively reduce drug release by 11 % in high pH environments and promote rapid drug release by 14 % in low pH environments compared to NRs without PDA. Additionally, F@Z/C/P can remove the biofilms and bacteria in Staphylococcus aureus infected wounds, and eventually be discharged from the infected site after treatment, leading to faster healing with an intact epidermis and minimal harm to surrounding tissues and organs. The study provides a promising strategy for tackling biofilm-associated infections in vivo through the use of multi-functional NRs.


Assuntos
Pandanaceae , Liberação Controlada de Fármacos , Frutas , Antibacterianos/química , Biofilmes , Cicatrização , Concentração de Íons de Hidrogênio , Catecóis/farmacologia
9.
IEEE Trans Pattern Anal Mach Intell ; 45(8): 9504-9519, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37021919

RESUMO

Effectively tackling the problem of temporal action localization (TAL) necessitates a visual representation that jointly pursues two confounding goals, i.e., fine-grained discrimination for temporal localization and sufficient visual invariance for action classification. We address this challenge by enriching the local, global and multi-scale contexts in the popular two-stage temporal localization framework. Our proposed model, dubbed ContextLoc++, can be divided into three sub-networks: L-Net, G-Net, and M-Net. L-Net enriches the local context via fine-grained modeling of snippet-level features, which is formulated as a query-and-retrieval process. Furthermore, the spatial and temporal snippet-level features, functioning as keys and values, are fused by temporal gating. G-Net enriches the global context via higher-level modeling of the video-level representation. In addition, we introduce a novel context adaptation module to adapt the global context to different proposals. M-Net further fuses the local and global contexts with multi-scale proposal features. Specially, proposal-level features from multi-scale video snippets can focus on different action characteristics. Short-term snippets with fewer frames pay attention to the action details while long-term snippets with more frames focus on the action variations. Experiments on the THUMOS14 and ActivityNet v1.3 datasets validate the efficacy of our method against existing state-of-the-art TAL algorithms.

10.
Acta Biomater ; 157: 566-577, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36481503

RESUMO

Wound healing from bacterial infections is one of the major challenges in the biomedical field. The traditional single administration methods are usually accompanied with side effects or unsatisfactory efficacy. Herein, we design dynamically evolving antibacterial and repair-promoting nanocomposites (NCs) by in situ self-assembling of zeolitic imidazolate framework-8 (ZIF-8) on the surface of barium titanate (BTO), and further loading with a small amount of ciprofloxacin (CIP). The new strategy of combining pH-stimulated drug delivery and ultrasound-controlled sonodyamics has the potential to dynamically evolve in infected wound sites, offering a multifunctional therapy. In vitro study demonstrates that the enhancement generation of reactive oxygen species through the sonodynamic process due to the heterostructures and a small amount of CIP released in an acidic environment are synergistically antibacterial, and the inhibition rate was >99.9%. In addition, reduced sonodynamic effect and Zn2+ generated along with the gradual degradation of ZIF-8 simultanously promote cell migration and tissue regeneration. The in vivo study of full-thickness skin wounds in mouse models demonstrate a healing rate of 99.3% could be achieved under the treatment of BTO@ZIF-8/CIP NCs. This work provides a useful improvement in rational design of multi-stimulus-responsive nanomaterials for wound healing. STATEMENT OF SIGNIFICANCE: A novel piezoelectric nanocomposite was proposed to realize sonodynamic therapy and pH-stimulated drug releasing simultaneously in wound healing treatment. The dynamically evolving structure of the piezoelectric nanocomposite in acidic microenvironment has been theoretically and experimentally verified to contribute to a continuous variation of sonodymanic strength, which accompanied with the gradual releasing of drug and biocompatible Zn2+effectively balanced antibacterial and repair-promoting effects. Both of the in vitro and in vivo study demonstrated that the strategy could significantly accelerate wound healing, inspiring researchers to optimize the design of multi-stimulus-responsive nanomaterials for various applications in biomedical and biomaterial fields.


Assuntos
Antibacterianos , Nanocompostos , Camundongos , Animais , Antibacterianos/uso terapêutico , Cicatrização , Nanocompostos/uso terapêutico , Nanocompostos/química , Sistemas de Liberação de Medicamentos , Materiais Biocompatíveis/farmacologia
11.
Talanta ; 255: 124212, 2023 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-36566558

RESUMO

Acinetobacter baumannii (A. baumannii) is a common pathogen that causes hospital-acquired infections and is resistant to a wide variety of antibiotics. Consequently, the rapid and highly sensitive detection of A. baumannii is required during the early stages of infection. Therefore, we developed a DNA-catalyzed amplification mechanism based on aptamers, combined with a novel fluorescence resonance energy transfer (FRET) method based on graphene oxide (GO), for the detection of A. baumannii. In the presence of A. baumannii, an aptamer bound to A. baumannii, releasing the template strand, which triggered an entropy-driven catalysis (EDC) reaction. One EDC product was then used as the catalyst for catalytic hairpin assembly (CHA) on a GO nanosheet. Finally, the GO released a huge amount of FAM-labeled DNA duplices, which could be detected with FRET. This strategy circumvented the extraction of nucleic acids and was easy to execute, with a detection time of ≤1.5 h. The detection of A. baumannii with this method ranges from 5 cfu/mL to 1 × 105 cfu/mL, with a detection limit of 1.1 cfu/mL. The method was sufficiently sensitive and specific to detect A. baumannii rapidly in cerebrospinal fluid. In summary, our strategy provides a new option for the early detection and point-of-care testing (POCT) of A. baumannii infections, allowing their earlier and more precise treatment.


Assuntos
Acinetobacter baumannii , Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Transferência Ressonante de Energia de Fluorescência/métodos , Acinetobacter baumannii/genética , DNA , Aptâmeros de Nucleotídeos/genética , Catálise , Técnicas Biossensoriais/métodos
12.
Front Pharmacol ; 14: 1260674, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38273822

RESUMO

Introduction: Acute myocardial infarction (AMI) is characterized by the loss of cardiomyocytes, which impairs cardiac function and eventually leads to heart failure. The induction of cardiomyocyte cell cycle activity provides a new treatment strategy for the repair of heart damage. Our previous study demonstrated that morroniside exerts cardioprotective effects. This study investigated the effects and underlying mechanisms of action of morroniside on cardiomyocyte cell cycle activity and cardiac repair following AMI. Methods: Neonatal rat cardiomyocytes (NRCMs) were isolated and exposed to oxygen-glucose deprivation (OGD) in vitro. A rat model of AMI was established by ligation of the left anterior descending coronary artery (LAD) in vivo. Immunofluorescence staining was performed to detect newly generated cardiomyocytes. Western blotting was performed to assess the expression of cell cycle-related proteins. Electrocardiography (ECG) was used to examine pathological Q waves. Masson's trichrome and wheat germ agglutinin (WGA) staining assessed myocardial fibrosis and hypertrophy. Results: The results showed that morroniside induced cardiomyocyte cell cycle activity and increased the levels of cell cycle proteins, including cyclin D1, CDK4, cyclin A2, and cyclin B1, both in vitro and in vivo. Moreover, morroniside reduced myocardial fibrosis and remodeling. Discussion: In conclusion, our study demonstrated that morroniside stimulates cardiomyocyte cell cycle activity and cardiac repair in adult rats, and that these effects may be related to the upregulation of cell cycle proteins.

13.
Front Pharmacol ; 13: 906933, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35754511

RESUMO

Background: Alcohol-associated liver disease (ALD) increases the health burden worldwide, but effective drugs to prevent ALD are lacking. Furfural is a small molecule that can limit alcohol production in microorganisms and may have the capacity to attenuate ethanol-induced toxicity. Methods: Human HepG2 cells were incubated with ethanol and furfural, and cell viability, NAD+/NADH ratio, and mitochondrial function assays were performed. RNA sequencing (RNA-seq) data were used to annotate enriched pathways, and these findings were confirmed by reverse transcription-quantitative PCR (RT-qPCR) and Western blotting. C57BL/6J mice were fed a Lieber-DeCarli liquid diet. After 4 weeks, biochemical analysis of mouse serum and histological analysis of mouse livers were performed. Results: Different concentrations of furfural exerted different effects on mitochondria: low-dose furfural reduced reactive oxygen species (ROS) production, maintained mitochondrial transmembrane potential, and inhibited apoptosis pathway activation, while high-dose furfural led to the opposite effects. In mice, furfural mitigated transaminase increases and attenuated the lipid metabolism disorder that had been induced by ethanol. Conclusion: Low-dose furfural reduced ethanol-induced toxicity in the liver. Consuming food or beverages containing the appropriate level of furfural when drinking alcohol may be a convenient and useful way to prevent ALD.

14.
Talanta ; 243: 123388, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35303554

RESUMO

Nucleic acid detection technology is now widely used in scientific research and clinical testing, such as infectious and genetic diseases screening, molecular diagnosis of tumors and pharmacogenomic research, which is also an important part of in vitro diagnostics (IVD). However, with the increasing requirements of diagnosis and treatment, existing nucleic acid detection technologies are facing challenges in dealing with the current problems (especially since the outbreak of coronavirus disease in 2019 (Covid-19)). Recently, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (CRISPR/Cas)-based diagnostics have become a hot spot of attention. CRISPR/Cas has been developed as a molecular detection tool besides scientific research in biology and medicine fields, and some CRISPR-based products have already been translated. It is known as the "next-generation molecular diagnostic technology" because of its advantages such as easy design and accurate identification. CRISPR/Cas relies on pre-amplification of target sequences and subsequent detection of Cas proteins. Combining the CRISPR/Cas system with various isothermal nucleic acid amplification strategies can generate amplified detection signals, enrich low abundance molecular targets, improve the specificity and sensitivity of analysis, and develop point-of-care (POC) diagnostic techniques. In this review, we analyze the current status of CRISPR/Cas systems and isothermal amplification, report the advantages of combining the two and summarize the recent progress with the integration of both technologies with POC sensors in the nucleic acid field. In addition, the challenges and future prospects of CRISPR technology combined with isothermal amplification strategies in biosensing and clinical applications are discussed.


Assuntos
COVID-19 , Ácidos Nucleicos , COVID-19/diagnóstico , Sistemas CRISPR-Cas/genética , Humanos , Técnicas de Amplificação de Ácido Nucleico , Sistemas Automatizados de Assistência Junto ao Leito
15.
Sci Rep ; 12(1): 22016, 2022 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-36539545

RESUMO

This study assessed the value of circRNAs (circular RNAs) as prognostic markers in BC (breast cancer). We searched pertinent studies on the PubMed, Embase, and Web of Science online databases published according to PRISMA guidelines. A random-effects model for meta-analysis was used to assess the combined effect size of the HRs (hazard ratios) of the included studies. The heterogeneity test used Cochran's Q-test and I2 statistics. Thirty of the 520 trials retrieved were included in the systematic review. A total of 11 chemotherapeutic agents were used in the included studies. A total of 30 studies on 30 circRNAs were included in the systematic review. Of the 30 relevant circRNAs, 28 were upregulated and two were downregulated in breast cancer versus normal samples, and both were associated with increased drug resistance. Nine of 30 studies were used for the meta-analysis. The results of the meta-analysis showed that the groups with circRNA upregulation and circRNA downregulation showed the same prognostic risk (HR = 1.37, 95% Cl: 0.80-2.36, I2 = 63.7%). The results of subgroup analysis showed that both upregulated circRNAs (HR = 2.24, 95% Cl: 1.34-3.75, I2 = 0%) and downregulated circRNAs (HR = 0.61, 95% Cl: 0.45-0.83, I2 = 0%) were associated with poor BC prognosis. Collectively, the results of all relevant articles collected indicated that circRNAs showed good potential as possible clinical biomarkers of chemoresistance in BC patients.


Assuntos
Neoplasias da Mama , RNA Circular , Humanos , Feminino , RNA Circular/genética , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Medicina de Precisão , Resistencia a Medicamentos Antineoplásicos/genética , Biomarcadores Tumorais/genética , Prognóstico
16.
Foods ; 11(16)2022 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-36010507

RESUMO

Acetamiprid (ACE) is widely used in various vegetables to control pests, resulting in residues and posing a threat to human health. For the rapid detection of ACE residues in vegetables, an indirect competitive chemiluminescence enzyme immunoassay (ic-CLEIA) was established. The optimized experimental parameters were as follows: the concentrations of coating antigen (ACE-BSA) and anti-ACE monoclonal antibody were 0.4 and 0.6 µg/mL, respectively; the pre-incubation time of anti-ACE monoclonal antibody and ACE (sample) solution was 30 min; the dilution ratio of goat anti-mouse-HRP antibody was 1:2500; and the reaction time of chemiluminescence was 20 min. The half-maximum inhibition concentration (IC50), the detection range (IC10-IC90), and the detection limit (LOD, IC10) of the ic-CLEIA were 10.24, 0.70-96.31, and 0.70 ng/mL, respectively. The cross-reactivity rates of four neonicotinoid structural analogues (nitenpyram, thiacloprid, thiamethoxam, and clothianidin) were all less than 10%, showing good specificity. The average recovery rates in Chinese cabbage and cucumber were 82.7-112.2%, with the coefficient of variation (CV) lower than 9.19%, which was highly correlated with the results of high-performance liquid chromatography (HPLC). The established ic-CLEIA has the advantages of simple pretreatment and detection process, good sensitivity and accuracy, and can meet the needs of rapid screening of ACE residues in vegetables.

17.
Foods ; 11(21)2022 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-36359944

RESUMO

Chlorpyrifos is an organophosphorus insecticide, which can be used to control a variety of chewing and piercing mouthparts pests in agricultural production. It can destroy the normal nerve impulse conduction by inhibiting the activity of acetylcholinesterase or cholinesterase in the nerves, causing a series of poisoning symptoms. In order to achieve the quantitative analysis of chlorpyrifos residues in agricultural products, an aptamer-controlled signal molecule release method was developed in this study. The signal molecule 4-ATP of surface-enhanced Raman spectroscopy (SERS) was loaded into aminated mesoporous silica nanoparticles (MSNs-NH2) prepared by the one pot method, and then coated with an aptamer of chlorpyrifos through electrostatic interaction. The specific binding of the aptamer and chlorpyrifos led to the release of 4-ATP, and the amount of 4-ATP released was positively correlated with the amount of chlorpyrifos. Finally, the standard curve of chlorpyrifos quantitative detection based on SERS was established. Meanwhile, Ag-carrying mesoporous silica (Ag@MSNs) was prepared as the reinforcement substrate for SERS detection. The results showed that there was a good linear correlation between the Raman intensity and the concentration of chlorpyrifos at 25−250 ng/mL, and the limit of detection (LOD) was 19.87 ng/mL. The recoveries of chlorpyrifos in the apple and tomato samples were 90.08−102.2%, with RSD < 3.32%. This method has high sensitivity, specificity, reproducibility and stability, and can be used for the quantitative detection of chlorpyrifos in the environment and agricultural products.

18.
Ann Transl Med ; 10(22): 1223, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36544689

RESUMO

Background: Chronic exposure to airborne microparticles has been shown to increase the incidence of several chronic diseases. Previous studies have found that waterfall forest aerosols contribute to a diminished immune stress response in patients with asthma. However, the specific effects of short-term waterfall forest aerosol exposure on lung proteins have not been fully elucidated. Methods: This study used liquid chromatography-tandem mass spectrometry (LC-MS) to analyze changes in protein expression in the lungs of rats exposed to short-term waterfall forest aerosol environments. Specific protein markers were identified using bioconductivity analysis screening and validated using immunohistochemistry. Results: Waterfall forest aerosol environment exposure on day 5 downregulated the expression of the classical inflammatory pathway nuclear factor κB (NF-κB) signaling pathway. As the waterfall forest aerosol environment increased due to the duration of exposure, it was involved in oxidative phosphorylation and then hormone signaling in lung cells from the very beginning. In contrast, at day 15 of exposure, there is an effect on the regulation of the immune-related high-affinity IgE receptor pathway. In addition, iron-sulfur Rieske protein (Uqcrfs1), mitochondrial Tu translation elongation factor (Tufm) and ribosomal protein L4 (Rpl4) were identified as possible bioindicators for the evaluation of air quality. Conclusions: These results provide a comprehensive proteomic analysis that supports the positive contribution of a good air quality environment to lung health.

19.
Ann Transl Med ; 10(16): 894, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36110995

RESUMO

Background: The aim of this study was to investigate the specific mechanisms underlying the human health-promoting effects of the forest environment at Huangguoshu Falls, Guizhou, China. Methods: Ninety-five participants were recruited and an eye tracker was used to record fixation and sweep indices. A questionnaire was also used to evaluate the effects of different subject environments on human emotions, perceived recovery and preferences. Thereafter, 24 participants with chronic fatigue syndrome (CFS) were recruited and the participants' fatigue and stress-related scale indices and inflammatory factor levels were examined. Serum metabolites of the participants under different time waterfall forest interventions were detected by ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UHPLC-Q/TOF-MS). Results: Eye tracking paradigm analysis showed that the "waterfall" element was the most interesting element for participants and that the "charm" of the waterfall forest environment could be well perceived by participants. Scores on the Fatigue Scale, Anxiety Scale and Depression Scale decreased as the duration of treatment in the waterfall forest environment increased. Levels of inflammatory factors decreased after treatment in the waterfall forest environment. At the same time the level of antioxidants, represented by L-ascorbic acid, increased significantly. Conclusions: The charm of the Huangguoshu waterfall scenery could be perceived by the participants and have a positive modulating effect on mood and cognitive function. In addition, the unique mixture of negative oxygen ions in this environment can increase the content of endogenous antioxidants and balance the metabolism of choline and amino acids.

20.
Front Oncol ; 12: 945143, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36419885

RESUMO

Background: Circular RNAs (circRNAs) are receiving increasing attention as novel biomarkers. Our goal was to investigate the diagnostic, clinicopathological, and prognostic utility of circRNAs in prostate cancer (PCa). Methods: Relevant literature was searched in PubMed, Web of Science, and EMBASE. Sensitivity, specificity, diagnostic odds ratio (DOR), negative likelihood ratio (NLR), positive likelihood ratio (PLR), and the area under the curve (AUC) were calculated to evaluate the diagnostic accuracy of circRNA expression. circRNAs' clinical, pathological, and prognostic value was examined using pooled odds ratios (ORs) and hazard ratios (HRs). Results: This meta-analysis included 23 studies, with 5 for diagnosis, 16 for clinicopathological parameters, and 10 for prognosis. For diagnostic value, the pooled sensitivity, pooled specificity, PLR, NLR, DOR, and AUC were 0.82, 0.62, 2.17, 0.29, 7.37, and 0.81, respectively. Upregulation of carcinogenic circRNAs was associated with poor clinical parameters (Gleason score: OR = 0.222, 95% CI: 0.145-0.340; T classification: OR = 0.274, 95% CI: 0.175-0.430; lymph node metastasis: OR = 0.353, 95% CI: 0.175-0.716; tumor size: OR = 0.226, 95% CI: 0.099-0.518) and could predict poor survival outcomes (HR = 2.408, 95% CI: 1.559-3.720, p < 0.001). Conversely, downregulation of tumor-suppressor circRNAs was also associated with poor clinical parameters (Gleason score: OR = 1.689, 95% CI: 1.144-2.493; T classification: OR = 2.586, 95% CI: 1.779-3.762) and worse prognosis (HR = 1.739, 95% CI: 1.147-2.576, p = 0.006). Conclusion: Our results showed that circRNAs might be useful biomarkers for the diagnosis and prognosis of PCa. Systematic review registration: https://www.crd.york.ac.uk/prospero/, identifier CRD42021284785.

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