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1.
Sensors (Basel) ; 21(14)2021 Jul 13.
Artículo en Inglés | MEDLINE | ID: mdl-34300506

RESUMEN

The sea area around Dajin Island in the Pearl River Estuary is the second-largest habitat in China for the Indo-Pacific humpback dolphin (Sousa Chinensis). However, the rapid economic development of this area brings potential threats to the aquatic ecology around Dajin Island. Real-time monitoring and evaluation of the ecological health of the sea area are urgent. In this study, band ratio and single-band inversion algorithms were performed to obtain Chlorophyll-a (Chl-a) and Suspended Sediment Concentration (SSC), relying on both Compact Airborne Spectrographic Imager (CASI) and Moderate resolution Imaging Spectrometer (MODIS) images. The CASI/Chl-a with high spatial resolution was adopted to assess the eutrophication status, while the dissolved organic carbon (DOC) concentration and chromophoric dissolved organic matter (CDOM) optical properties were used to derive the material composition and sources. The results suggest that the study area is under a low to medium eutrophication state with evenly distributed low Chl-a concentration. However, higher Chl-a is observed in the outer estuary with MODIS/Chl-a. The relatively high DOC concentration, especially in the north, where aquaculture is practiced, and near the estuary's main axis, i.e., east Dajin Island, indicates that the eutrophication state might be underestimated using satellite chlorophyll alone. CDOM optical properties indicated that terrestrial materials are the DOC's primary material sources, but the DOC derived from fishery aquaculture cannot be ignored. The low Chl-a concentration is likely due to the turbulent hydrodynamic regime caused by jet flow and reciprocating flow in this marine area. Comprehensive observation, including the assessment of different technological platforms, is suggested for the aquatic environment.


Asunto(s)
Carbono , Monitoreo del Ambiente , Carbono/análisis , China , Clorofila A , Eutrofización
2.
Mikrochim Acta ; 187(3): 160, 2020 02 10.
Artículo en Inglés | MEDLINE | ID: mdl-32040773

RESUMEN

An indirect aptamer-based SERS assay for insulin-like growth factor 2 receptor (IGF-IIR) protein was developed. The gold substrate and silver nanoparticles (AgNPs) were employed simultaneously to achieve double enhancement for SERS signals. Firstly, the five commercial SERS substrates including Enspectr, Ocean-Au, Ocean-AG, Ocean-SP and Q-SERS substrates were evaluated using 4-mercaptobenzoic acid (4-MBA). The Q-SERS substrate was selected based on low relative standard deviation (RSD, 8.6%) and high enhancement factor (EF, 8.7*105), using a 785 nm laser. The aptamer for IGF-IIR protein was designed to include two sequences: one grafted on gold substrate to specifically capture the IGF-IIR protein and a second one forming a 3' sticky bridge to capture SERS nanotags. The SERS nanotag was composed by AgNPs (20 nm), 4-MBA and DNA probes that can hybridize with the aptamer. Due to the steric-hindrance effect, when the aptamer doesn't combine with IGF-IIR protein, it only can capture the SERS nanotags. Therefore, there was a negative correlation between the concentration of IGF-IIR protein and the intensity of 4-MBA at 1076 cm-1. The detection limit reached to 141.2 fM and linear range was from 10 pM to 1 µM. The SERS aptasensor also exhibits a high reproducibility with an average RSD of 4.5%. The interference test was conducted with other four proteins to verify the accuracy of measuring. The study provides an approach to quantitative determination of proteins based on specific recognition and nucleic acid hybridization of aptamers, to establish sandwich structure for SERS enhancement. Graphical abstractSchematic representation of surface-enhanced Raman scattering (SERS) assay on insulin-like growth factor 2 receptor (IGF-IIR) protein by combining the aptamer modified gold substrate and 4-mercaptobenzoic acid (4-MBA) and DNA probe modified silver nanoparticles.


Asunto(s)
Aptámeros de Nucleótidos/química , Oro/química , Nanopartículas del Metal/química , Receptor IGF Tipo 2/genética , Plata/química , Espectrometría Raman/métodos
3.
Tumour Biol ; 35(9): 8837-48, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24880590

RESUMEN

Dysregulated microRNAs (miRNAs) have been reported to be associated with pancreatic cancer (PaC), suggesting that they may serve as useful novel diagnostic biomarkers for PaC. Various studies have been performed to investigate the diagnostic value of miRNAs for PaC but have obtained conflicting results. Therefore, this meta-analysis aims to comprehensively and quantitatively evaluate the potential diagnostic value of miRNAs for PaC. We systematically searched PubMed, Embase, Google Scholar, Cochrane Library, and Chinese National Knowledge Infrastructure for publications concerning the diagnostic value of miRNAs for PaC without language restriction. The quality of each study was scored using the revised Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2). The summary receiver operator characteristic curve and other parameters were applied to check the overall test performance. Heterogeneity was tested with the I (2) test and publication bias was tested with the Deek's funnel plot asymmetry test. This meta-analysis included 18 articles with a total of 2,036 patients and 1,444 controls. The pooled sensitivity was 82 % (95 % CI, 78-86 %); the specificity was 77 % (95 % CI, 73-81 %); the PLR was 3.6 (95 % CI, 3.0-4.4); the NLR was 0.23 (95 % CI, 0.18-0.29); the DOR was 16 (95 % CI, 10-24); and the AUC was 0.86 (95 % CI, 0.83-0.89). Subgroups analyses were also performed and revealed that there were significant differences between some subgroups: the multiple-miRNAs profiling-based assays, non-blood-based assays, and healthy control-based studies all showed higher accuracies in diagnosing PaC than that of their counterparts. This meta-analysis suggests that the use of miRNAs has potential diagnostic value with a relatively high sensitivity and specificity for PaC, particularly the use of multiple miRNAs for discriminating PaC patients from healthy individuals. More prospective studies on the diagnostic value of miRNAs for PaC are needed in the future.


Asunto(s)
Biomarcadores de Tumor/genética , MicroARNs/genética , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/genética , Diagnóstico Diferencial , Humanos , Sensibilidad y Especificidad
4.
Talanta ; 272: 125760, 2024 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-38364563

RESUMEN

Staphylococcus aureus (S. aureus) poses a serious threat to global public health, necessitating the establishment of rapid and simple tools for its accurate identification. Herein, we developed a terahertz (THz) metamaterial biosensor based on aptamer-functionalized Fe3O4@Au nanocomposites for quantitative S. aureus assays in different clinical samples. Fe3O4@Au@Cys@Apt has the dual advantages of magnetism and a high refractive index in the THz range and was used to rapidly separate and enrich target bacteria in a complex environmental solution. Furthermore, conjugation to the nanocomposites significantly increased the resonance frequency shift of the THz metamaterial after target loading. Our results showed that the shifts in the metamaterial resonance frequency were linearly related to S. aureus concentrations ranging from 1.0 × 103 to 1.0 × 107 CFU/mL, with a detection limit of 4.78 × 102 CFU/mL. The biosensor was further applied to S. aureus detection in spiked human urine and blood with satisfactory recoveries (82.4-109.6%). Our approach also demonstrated strong concordance with traditional plate counting (R2 = 0.99306) while significantly lowering the analysis time from 24 h to <1 h. In conclusion, the proposed biosensor can not only perform culture-free and extraction-free detection of target bacteria but can also be easily extended to the determination of other pathogenic bacteria, rendering it suitable for various bacteria-related disease diagnoses.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Nanocompuestos , Infecciones Estafilocócicas , Humanos , Staphylococcus aureus , Técnicas Biosensibles/métodos , Infecciones Estafilocócicas/diagnóstico , Infecciones Estafilocócicas/microbiología , Bacterias , Oro
5.
ACS Sens ; 9(4): 2122-2133, 2024 04 26.
Artículo en Inglés | MEDLINE | ID: mdl-38602840

RESUMEN

Terahertz (THz) spectroscopy has impressive capability for label-free biosensing, but its utility in clinical laboratories is rarely reported due to often unsatisfactory detection performances. Here, we fabricated metal-graphene hybrid THz metasurfaces (MSs) for the sensitive and enzyme-free detection of circulating tumor DNA (ctDNA) in pancreatic cancer plasma samples. The feasibility and mechanism of the enhanced effects of a graphene bridge across the MS and amplified by gold nanoparticles (AuNPs) were investigated experimentally and theoretically. The AuNPs serve to boost charge injection in the graphene film and result in producing a remarkable change in the graded transmissivity index to THz radiation of the MS resonators. Assay design utilizes this feature and a cascade hybridization chain reaction initiated on magnetic beads in the presence of target ctDNA to achieve dual signal amplification (chemical and optical). In addition to demonstrating subfemtomolar detection sensitivity and single-nucleotide mismatch selectivity, the proposed method showed remarkable capability to discriminate between pancreatic cancer patients and healthy individuals by recognizing and quantifying targeted ctDNAs. The introduction of graphene to the metasurface produces an improved sensitivity of 2 orders of magnitude for ctDNA detection. This is the first study to report the combined application of graphene and AuNPs in biosensing by THz spectroscopic resonators and provides a combined identification scheme to detect and discriminate different biological analytes, including nucleic acids, proteins, and various biomarkers.


Asunto(s)
ADN Tumoral Circulante , Oro , Grafito , Nanopartículas del Metal , Neoplasias Pancreáticas , Grafito/química , Humanos , Oro/química , Nanopartículas del Metal/química , ADN Tumoral Circulante/sangre , ADN Tumoral Circulante/genética , ADN Tumoral Circulante/análisis , Neoplasias Pancreáticas/sangre , Neoplasias Pancreáticas/diagnóstico , Técnicas Biosensibles/métodos , Espectroscopía de Terahertz/métodos , Hibridación de Ácido Nucleico , Límite de Detección
6.
Tumour Biol ; 34(2): 1021-6, 2013 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-23283818

RESUMEN

This study aims to investigate the expression and significance of KPNA2 in human gastric adenocarcinoma progression and prognosis. Using immunohistochemistry and real-time reverse transcriptase polymerase chain reaction assay, we identified abnormally elevated expression of KPNA2 in gastric adenocarcinoma tissues compared to paired normal stomach mucosa tissues in 30 patients (p < 0.05). In order to investigate the correlations between KPNA2 and the clinicopathological features of gastric adenocarcinoma, the expression of KPNA2 in 142 patients with gastric adenocarcinoma was detected by immunohistochemistry, and the results showed that overexpression of KPNA2 was associated with the size of tumor (p < 0.001), histological grade (p < 0.001), lymph node involvement (p = 0.001), and tumor node metastasis stage (p < 0.001). Kaplan-Meier survival analysis showed that patients with high KPNA2 expression showed a significantly shorter overall survival time compared with patients with low KPNA2 expression. Multivariate analysis suggested that KPNA2 expression might be an independent prognostic indicator (p < 0.001) for the survival of patients with gastric adenocarcinoma. In conclusion, overexpression of KPNA2 is closely related to progression of gastric adenocarcinoma and might be regarded as an independent predictor of poor prognosis for gastric adenocarcinoma.


Asunto(s)
Adenocarcinoma Mucinoso/mortalidad , Adenocarcinoma/mortalidad , Carcinoma Papilar/mortalidad , Carcinoma de Células en Anillo de Sello/mortalidad , Neoplasias Gástricas/mortalidad , alfa Carioferinas/metabolismo , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Adenocarcinoma Mucinoso/metabolismo , Adenocarcinoma Mucinoso/patología , Adulto , Anciano , Carcinoma Papilar/metabolismo , Carcinoma Papilar/patología , Carcinoma de Células en Anillo de Sello/metabolismo , Carcinoma de Células en Anillo de Sello/patología , Estudios de Casos y Controles , Femenino , Estudios de Seguimiento , Humanos , Técnicas para Inmunoenzimas , Ganglios Linfáticos/patología , Metástasis Linfática , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Pronóstico , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patología , Tasa de Supervivencia , Adulto Joven , alfa Carioferinas/genética
7.
Mol Cell Biochem ; 383(1-2): 201-11, 2013 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-23934089

RESUMEN

Bone marrow stromal stem cells (BMSCs) are fibroblastic in shape and capable of self-renewal and have the potential for multi-directional differentiation. Nerve growth factor (NGF), a homodimeric polypeptide, plays an important role in the nervous system by supporting the survival and growth of neural cells, regulating cell growth, promoting differentiation into neuron, and neuron migration. Adenoviral vectors are DNA viruses that contain 36 kb of double-stranded DNA allowing for transmission of the genes to the host nucleus but not inserting them into the host chromosome. The present study aimed to investigate the induction efficiency and differentiation of neural cells from BMSCs by ß-NGF gene transfection with recombinant adenoviral vector (Ad-ß-NGF) in vitro. The results of immunochemical assay confirmed the induced cells as neuron cells. Moreover, flow cytometric analysis, Annexin-V-FITC/PI, and BrdU assay revealed that chemical inducer ß-mercaptoethanol (ß-met) triggered apoptosis of BMSCs, as evidenced by inhibition of DNA fragmentation, nuclear condensation, translocation of phospholipid phosphatidylserine, and activation of caspase-3. Furthermore, the results of western blotting showed that ß-met suppressed AKT signaling pathway and regulated the MAPKs during differentiation of BMSCs. In contrast, Ad-ß-NGF effectively induced the differentiation of BMSCs without causing any cytopathic phenomenon and apoptotic cell death. Moreover, Ad-ß-NGF recovered the expression level of phosphorylated AKT and MAPKs in cells exposed to chemical reagents. Taken together, these results suggest that ß-NGF gene transfection promotes the differentiation of BMSCs into neurons through regulation of AKT and MAPKs signaling pathways.


Asunto(s)
Diferenciación Celular , Sistema de Señalización de MAP Quinasas , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/enzimología , Factores de Crecimiento Nervioso/metabolismo , Neuronas/citología , Precursores de Proteínas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Adenoviridae/genética , Animales , Apoptosis/efectos de los fármacos , Células de la Médula Ósea/citología , Caspasa 3/metabolismo , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Forma de la Célula/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Vectores Genéticos/metabolismo , Inmunohistoquímica , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Masculino , Mercaptoetanol/farmacología , Células Madre Mesenquimatosas/metabolismo , Ratones , Neuronas/efectos de los fármacos , Neuronas/enzimología , Fosforilación/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Transfección
8.
Tohoku J Exp Med ; 229(4): 255-66, 2013 04.
Artículo en Inglés | MEDLINE | ID: mdl-23603420

RESUMEN

High-altitude pulmonary edema (HAPE) is a non-cardiogenic type of pulmonary edema developing altitudes > 2,500 m. Angiotensin converting enzyme (ACE) and nitric oxide synthase 3 (NOS3) play important roles in regulating pulmonary vascular tone. To assess associations between genetic variants in the ACE and NOS3 genes and HAPE risk, 27 HAPE patients and 108 matched controls were genotyped and analyzed. The indicated HAPE association of the NOS3 G894T (Glu298Asp) single nucleotide polymorphism (SNP), which may change NO production, was further evaluated by a meta-analysis of six studies involving 399 HAPE patients and 495 controls. Odds ratios (ORs) and 95% confidence intervals (CIs) were determined with fixed-effects models. Stratification analyses of ethnicity and geographic location were performed. Significant associations were observed for the dominant model in two ACE tag SNPs influencing serum ACE concentrations (rs8066114 polymorphism: GG+CG vs. CC; rs4461142 polymorphism: TT+CT vs. CC). Furthermore, Single-locus analysis indicated significantly different distributions of G allele frequency between the cases (29.63%) and controls (17.13%) for the ACE rs8066114 polymorphism. The case-control distributions of genotype frequencies and T allele frequency of NOS3 G894T (Glu298Asp) polymorphism were significantly higher in the cases than controls, and the NOS3 G894T (Glu298Asp) SNP showed elevated HAPE risk under the dominant model (TT+GT vs. GG). Meta-analysis showed overall association of NOS3 G894T SNP with HAPE risk under the allele contrast and dominant genetic models, which remained significant for Asians. In conclusion, ACE rs8066114 and rs4461142 and NOS3 rs1799983 (G894T) polymorphisms may be associated with increased HAPE risk in Asians.


Asunto(s)
Mal de Altura/genética , Hipertensión Pulmonar/genética , Óxido Nítrico Sintasa de Tipo III/genética , Peptidil-Dipeptidasa A/genética , Polimorfismo de Nucleótido Simple/genética , Mal de Altura/etnología , Pueblo Asiatico/genética , Estudios de Casos y Controles , China , Cartilla de ADN/genética , Estudios de Asociación Genética , Genotipo , Humanos , Hipertensión Pulmonar/etnología , Modelos Genéticos , Oportunidad Relativa , Reacción en Cadena de la Polimerasa , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
9.
Biosensors (Basel) ; 13(10)2023 Oct 23.
Artículo en Inglés | MEDLINE | ID: mdl-37887140

RESUMEN

The sensitive and accurate detection of tumor cells is essential for successful cancer therapy and improving cancer survival rates. However, current tumor cell detection technologies have some limitations for clinical applications due to their complexity, low specificity, and high cost. Herein, we describe the design of a terahertz anti-resonance hollow core fiber (THz AR-HCF) biosensor that can be used for tumor cell detection. Through simulation and experimental comparisons, the low-loss property of the THz AR-HCF was verified, and the most suitable fiber out of multiple THz AR-HCFs was selected for biosensing applications. By measuring different cell numbers and different types of tumor cells, a good linear relationship between THz transmittance and the numbers of cells between 10 and 106 was found. Meanwhile, different types of tumor cells can be distinguished by comparing THz transmission spectra, indicating that the biosensor has high sensitivity and specificity for tumor cell detection. The biosensor only required a small amount of sample (as low as 100 µL), and it enables label-free and nondestructive quantitative detection. Our flow cytometry results showed that the cell viability was as high as 98.5 ± 0.26% after the whole assay process, and there was no statistically significant difference compared with the negative control. This study demonstrates that the proposed THz AR-HCF biosensor has great potential for the highly sensitive, label-free, and nondestructive detection of circulating tumor cells in clinical samples.


Asunto(s)
Técnicas Biosensibles , Neoplasias , Humanos , Fibras Ópticas , Simulación por Computador , Tecnología
10.
Biosensors (Basel) ; 12(6)2022 May 31.
Artículo en Inglés | MEDLINE | ID: mdl-35735526

RESUMEN

The demand for rapid and accurate identification of microorganisms is growing due to considerable importance in all areas related to public health and safety. Here, we demonstrate a rapid and label-free strategy for the identification of microorganisms by integrating terahertz-attenuated total reflection (THz-ATR) spectroscopy with an automated recognition method based on multi-classifier voting. Our results show that 13 standard microbial strains can be classified into three different groups of microorganisms (Gram-positive bacteria, Gram-negative bacteria, and fungi) by THz-ATR spectroscopy. To detect clinical microbial strains with better differentiation that accounts for their greater sample heterogeneity, an automated recognition algorithm is proposed based on multi-classifier voting. It uses three types of machine learning classifiers to identify five different groups of clinical microbial strains. The results demonstrate that common microorganisms, once time-consuming to distinguish by traditional microbial identification methods, can be rapidly and accurately recognized using THz-ATR spectra in minutes. The proposed automatic recognition method is optimized by a spectroscopic feature selection algorithm designed to identify the optimal diagnostic indicator, and the combination of different machine learning classifiers with a voting scheme. The total diagnostic accuracy reaches 80.77% (as high as 99.6% for Enterococcus faecalis) for 1123 isolates from clinical samples of sputum, blood, urine, and feces. This strategy demonstrates that THz spectroscopy integrated with an automatic recognition method based on multi-classifier voting significantly improves the accuracy of spectral analysis, thereby presenting a new method for true label-free identification of clinical microorganisms with high efficiency.


Asunto(s)
Algoritmos , Bacterias/clasificación , Hongos/clasificación , Interacciones Microbiota-Huesped , Espectroscopía de Terahertz , Aprendizaje Automático , Salud Pública , Seguridad , Análisis Espectral , Espectroscopía de Terahertz/métodos , Virus/clasificación
11.
Dis Markers ; 2022: 8241405, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35299867

RESUMEN

Objective: This study is aimed at investigating the early diagnosis and efficacy of emergency treatments of nine patients with severe multiple injuries accompanied by traumatic aortic dissection (TAD). Methods: Patients who sustained severe multiple injuries accompanied by TAD following a car accident (n = 6) and falls from a height (n = 3) were treated in the emergency department of our hospital from October 2017 to July 2021. Data of these patients, including seven men and two women (average age, 53 ± 15.2 years; range, 18-83 years) were analysed retrospectively. Upon hospital arrival, the multidisciplinary treatment (MDT) trauma team, composed of doctors and nurses, immediately performed resuscitation following the Green Channel Consultation and Treatment Process for Severe Multiple Injuries. Life-threatening injuries were managed urgently. Blood tests and blood preparation and bedside B-scan ultrasonography and CT were performed. Aortic computed tomography angiography (CTA) was conducted decisively in patients suspected of TAD so that endovascular graft exclusion (EVGE) with the aortic covered stent can be performed promptly, followed by emergency management, second-stage surgery, and intensive care according to the injury control strategy. Results: This study included nine patients suffering from severe multiple injuries accompanied by Stanford type B TAD, with injury severity scores ranging from 35 to 43 points. Six patients underwent EVGE while receiving emergency treatment, whereas two patients who also had intracranial haemorrhage underwent selective EVGE. One case of TAD missed in the emergency department was detected 13 days after hospitalisation; therefore, the patient promptly underwent EVGE. Emergency procedures performed included exploratory laparotomy and splenectomy (n = 2), thoracic closed drainage (n = 5), haemothoracotomy (n = 3), second-stage fracture surgery (n = 4), and tracheotomy (n = 1). Postinjury complications included haemorrhagic shock, coagulation disorders, hyoxaemia, pulmonary infection, renal insufficiency, and hypoproteinaemia; however, all patients recovered after intensive care treatment. Aortic CTA after EVGE revealed the disappearance of the dissection and the resorption of the intermural haematoma. However, varying degrees of stenosis or occlusion were observed in the left subclavian artery. Nine patients with severe multiple injuries were treated satisfactorily by the MDT, without fatalities, and all patients were discharged for rehabilitation. Conclusion: In this study, procedures including resuscitation, urgent aortic CTA for definitive diagnosis, prompt EVGE, emergency injury control surgery, second-stage definitive surgery, intensive care treatment, and rehabilitation were rationally performed by the emergency MDT trauma team. Overall, this continuous and seamless process is a key factor for the successful treatment of patients with severe multiple injuries accompanied by TAD.


Asunto(s)
Aorta Torácica/cirugía , Disección Aórtica/cirugía , Diagnóstico Precoz , Tratamiento de Urgencia , Traumatismo Múltiple/terapia , Stents , Angiografía por Tomografía Computarizada , Cuidados Críticos , Procedimientos Endovasculares , Femenino , Humanos , Puntaje de Gravedad del Traumatismo , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Tomografía Computarizada por Rayos X , Trasplantes , Resultado del Tratamiento
12.
Anal Chim Acta ; 1203: 339706, 2022 Apr 22.
Artículo en Inglés | MEDLINE | ID: mdl-35361424

RESUMEN

Sensitive and specific detection of microRNAs (miRNAs) is of critical significance for early diagnosis of cancers such as pancreatic cancer with atypical initial symptoms and high mortality. Despite exponential amplification reaction (EXPAR) is an attractive isothermal amplification method for detecting miRNAs, it faces the problems of the dependence difference and low specificity. To address such challenges, herein, a nicking-assisted entropy-driven DNA circuit triggered exponential amplification reaction (NAED-EXPAR) was firstly employed for ultrasensitive and specific detection of miRNA in "one-pot" manner at constant temperature. Nicking-assisted entropy-driven DNA circuit can specifically recognize the target miRNA, leading to continuous disassembly of DNA substrates via intramolecular toehold-mediated branch migration. During the reaction, the catalytic circuit can consume excess fuel DNA strands to produce a large number of primers. Then the newly formed primers can trigger EXPAR for highly efficient signal amplification. Mechanism analysis shows that the amplification efficiency of NAED-EXPAR is superior than that of single EXPAR. For miR-21, the detection limit of NAED-EXPAR can reach 100 aM, which is at least five orders of magnitude higher than the standard EXPAR that directly uses the target as primer. NAED-EXPAR shows improved specificity for identifying single nucleotide variations and enables sensitive and accurate analysis of miR-21 in human cancer cell lines. This method is expected to offer a new approach for the reliable quantification of miRNAs in complex biological matrices and provide valuable information for early cancer diagnosis.


Asunto(s)
MicroARNs , Neoplasias , ADN/química , ADN/genética , Entropía , Humanos , MicroARNs/análisis , MicroARNs/genética , Técnicas de Amplificación de Ácido Nucleico/métodos
13.
Talanta ; 228: 122213, 2021 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-33773748

RESUMEN

Terahertz (THz) spectroscopy, with fascinating advantages for biomedical applications, is still in its infancy in terms of the selective detection of aqueous biomolecules because the strong absorption of solvent water always obscures the THz spectroscopic features of biomolecules. Nevertheless, solvent water is not a passive spectator but a useful indicator, as this proposed strategy describes. This strategy utilizes THz attenuated total reflection (THz-ATR) spectroscopy to probe the glucose-induced hydration state changes of smart hydrogels for label-free and selective detection of aqueous glucose. A notable dramatic increase in both the THz absorption coefficient and hydration state (calculated by weighing) of the smart hydrogel was observed with increasing aqueous glucose concentration, which was further verified by a simple two-component model. For aqueous glucose sensing, this method surpasses individual THz-ATR devices and exhibits suitable sensitivity, ideal selectivity and excellent reusability. Moreover, the proposed strategy may provide an alternative option for the selective detection of various aqueous molecules by THz spectroscopy.

14.
ACS Sens ; 6(5): 1884-1890, 2021 05 28.
Artículo en Inglés | MEDLINE | ID: mdl-33979138

RESUMEN

Metamaterial-inspired terahertz (THz) biosensors are devoted to developing high-sensitivity and label-free biosensing strategies. However, most meaningful molecular signals are obscured by the strong THz absorption of solvent water. Most reported THz biosensors require the tested samples to be tediously dried or replaced with a low-absorption medium, which impairs the original bioactivity and the distribution homogeneity of targets. As described in this proposed strategy, a molecule-specific THz biosensor was fabricated from an aptamer hydrogel-functionalized THz metamaterial. Benefitting from the strong interaction with the localized electric field of the metamaterial, trace thrombin-induced variations in the hydration state of the hydrogel can be sensitively probed, which was investigated experimentally and theoretically. The optimized THz biosensor exhibited remarkable specificity for actual serum sample assays and excellent sensitivity, with a relatively low detection limit of 0.40 pM in the human serum matrix. The proposed strategy could serve as a model system to develop various molecule-specific THz biosensors for aqueous molecule sensing.


Asunto(s)
Técnicas Biosensibles , Hidrogeles , Bioensayo , Humanos , Agua
15.
Front Oncol ; 11: 665176, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34646758

RESUMEN

Gastric cancer (GC) is the fifth most common cancer in the world and a serious threat to human health. Due to its high morbidity and mortality, a simple, rapid and accurate early screening method for GC is urgently needed. In this study, the potential of Raman spectroscopy combined with different machine learning methods was explored to distinguish serum samples from GC patients and healthy controls. Serum Raman spectra were collected from 109 patients with GC (including 35 in stage I, 14 in stage II, 35 in stage III, and 25 in stage IV) and 104 healthy volunteers matched for age, presenting for a routine physical examination. We analyzed the difference in serum metabolism between GC patients and healthy people through a comparative study of the average Raman spectra of the two groups. Four machine learning methods, one-dimensional convolutional neural network, random forest, support vector machine, and K-nearest neighbor were used to explore identifying two sets of Raman spectral data. The classification model was established by using 70% of the data as a training set and 30% as a test set. Using unseen data to test the model, the RF model yielded an accuracy of 92.8%, and the sensitivity and specificity were 94.7% and 90.8%. The performance of the RF model was further confirmed by the receiver operating characteristic (ROC) curve, with an area under the curve (AUC) of 0.9199. This exploratory work shows that serum Raman spectroscopy combined with RF has great potential in the machine-assisted classification of GC, and is expected to provide a non-destructive and convenient technology for the screening of GC patients.

16.
Biosens Bioelectron ; 188: 113314, 2021 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-34030095

RESUMEN

Exosomal microRNA (miRNA) is a promising non-invasive biomarker for liquid biopsies. Herein, we fabricated a terahertz (THz) metamaterial biosensor that comprises an array of gold (Au) discs surrounded by annular grooves for exosomal miRNA assays based on duplex-specific nuclease (DSN)-triggered rolling circle amplification (RCA). In this strategy, the target miRNA is captured by a probe P0 immobilized on magnetic beads (MBs); it then repeatedly releases a primer P1 under the action of DSN, which acts as a highly specific initiator of the subsequent RCA step utilizing biotin-dUTP. After target recycling and nucleic acid amplification, the biotinylated amplification products were captured by the streptavidin (SA)-functionalized THz metamaterials, and further conjugated to SA-modified AuNPs that permit formation of a trimeric complex of SA-biotinylated RCA products-AuNP. The complex population scales with the starting concentration of the target miR-21, resulting in a red shift of the resonance peak of the THz metamaterials. This biosensor can lead to highly specific and sensitive detection with one-base mismatch discrimination and a limit of detection (LOD) down to 84 aM. Significant distinctions are seen in the frequency shifts for exosomal miR-21 quantitation in clinical plasma samples between pancreatic cancer patients and healthy controls. The frequency shifts of the THz metamaterials are consistent versus the reverse transcription-polymerase chain reaction (RT-PCR) results, illustrating the applicability and accuracy of our assay in real clinical samples.


Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , MicroARNs , Neoplasias Pancreáticas , Oro , Humanos , Límite de Detección , Técnicas de Amplificación de Ácido Nucleico , Neoplasias Pancreáticas/genética , Estreptavidina
17.
Biosens Bioelectron ; 175: 112874, 2021 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-33293192

RESUMEN

Terahertz (THz) spectroscopy has drawn great interest for the functional and conformational investigations of nucleic acids, but its intrinsic sensitivity hinders potential bio-sensing applications. Here, a novel THz biosensor was developed for detecting microRNA (miRNA) samples based on metamaterials coupled with nanoparticles and strand displacement amplification (SDA). In this method, the SDA reaction amplifies the target miRNA and generates copious yields of secondary DNA molecules (Trigger DNA), which are subsequently conjugated to metallic nanoparticles that form nanoparticle-Trigger DNA complexes. These complexes produce remarkable frequency shifts of metamaterials when linked to a large refractive index metallic nanoparticle like Au. The dependence of the metamaterial resonance on the nanoparticle diameter and metal type was investigated experimentally and theoretically. Under optimal conditions, the THz metamaterial biosensor presents good detection sensitivity with a limit of detection of 14.54 aM and exhibits a linear response for miRNA-21 at a concentration range from 1 fM to 10 pM. By measuring the miRNA-21 in spiked clinical serum samples, the sample recoveries were determined to be in the range between 90.92% and 107.01%. These findings demonstrate that the novel THz biosensor offers the capability for highly sensitive miRNA detection, with noteworthy potential applications in nucleic acid analysis and cancer diagnosis.


Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , MicroARNs , ADN/genética , Oro , Límite de Detección , Técnicas de Amplificación de Ácido Nucleico
18.
Breast Cancer Res Treat ; 122(2): 509-14, 2010 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-20052533

RESUMEN

Breast cancer is the most common cancer in women worldwide, but its etiology is still unclear. It is believed that oxidative stress plays an essential role in the development of breast cancer, while SOD2 is one of the primary enzymes that directly convert potential harmful oxidizing species to harmless metabolites. The association of SOD2 Val16Ala polymorphism and breast cancer risk has been widely reported, but results of previous studies were somewhat contradictory and underpowered. To overcome the limitations of individual study and to understand the real situation, we conducted a systematic review and meta-analysis toward the association between SOD2 Val16Ala polymorphism and breast cancer. Through retrieving MEDLINE, PubMed, Embase, and Web of Science, a total of 17 studies with 9,710 cases and 11,041 controls were identified. The results showed that no significant associations were found for the allele contrast (allele Ala vs. allele Val: OR = 1.020, 95% CI = 0.979-1.062), additive genetic model (Ala/Ala vs. Val/Val: OR = 1.091, 95% CI = 0.969-1.229), dominant genetic model (Ala/Ala +Ala/Val vs. Val/Val: OR = 1.045, 95% CI = 0.961-1.136), and recessive genetic model (Ala/Ala vs. Val/Val +Ala/Val: OR = 1.027, 95% CI = 0.956-1.102). In the stratified analysis by ethnicity and menopausal status, significant associations were also not detected in all genetic models. Conclusively, this meta-analysis strongly suggests that SOD2 Val16Ala polymorphism is not associated with breast cancer susceptibility.


Asunto(s)
Neoplasias de la Mama/genética , Polimorfismo Genético , Superóxido Dismutasa/genética , Neoplasias de la Mama/enzimología , Estudios de Casos y Controles , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Humanos , Oportunidad Relativa , Fenotipo , Medición de Riesgo , Factores de Riesgo
19.
Virol J ; 7: 255, 2010 Sep 28.
Artículo en Inglés | MEDLINE | ID: mdl-20875143

RESUMEN

BACKGROUND: Bacteriophages (phages) are widespread in the environment and play a crucial role in the evolution of their bacterial hosts and the emergence of new pathogens. RESULTS: LSB-1, a reference coliphage strain, was classified as a member of the Podoviridae family with a cystic form (50 ± 5 nm diameter) and short tail (60 ± 5 nm long). The double stranded DNA was about 30 kilobase pairs in length. We identified its host range and determined the gp17 sequences and protein structure using shotgun analysis and bioinformatics technology. CONCLUSIONS: Coliphage LSB-1 possesses a tailspike protein with endosialidase activity which is probably responsible for its specific enteroinvasive E.coli host range within the laboratory.


Asunto(s)
Colifagos/genética , Colifagos/fisiología , Especificidad del Huésped , Podoviridae/genética , Podoviridae/fisiología , Secuencia de Aminoácidos , Colifagos/clasificación , Colifagos/ultraestructura , Biología Computacional , ADN Viral/química , ADN Viral/genética , Escherichia coli/virología , Modelos Moleculares , Datos de Secuencia Molecular , Filogenia , Podoviridae/clasificación , Podoviridae/ultraestructura , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Análisis de Secuencia de ADN , Proteínas Virales/genética , Virión/ultraestructura
20.
RSC Adv ; 10(45): 26824-26833, 2020 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-35515811

RESUMEN

A highly sensitive method for detecting Staphylococcus aureus (S. aureus) is urgently needed to reduce the impact and spread of hospital-acquired infections and food-borne illness. For this purpose, this paper presents a THz metamaterial biosensor based on gold nanoparticles (AuNPs) and rolling circle amplification (RCA). The RCA process amplified the S. aureus DNA fragments and generated copious yields of long single-strand DNA molecules. These molecules were then conjugated with the AuNPs to form complexes that delivered exceptional increases in the refractive indices of the samples, and resulted in corresponding improvements in the THz response of the metamaterial. Under optimal conditions, the shifts in the metamaterial's resonance frequency displayed a linear relationship with concentrations of synthetic S. aureus DNA in the range from 10 fM to 10 pM, with a limit of detection of 2.77 fM. We also tested the practical application of this biosensor in measurements of genomic DNA in clinical bacterial strains, where the sensor showed a detection limit of 0.08 pg µL-1 and a linear range from 0.1 to 5 pg µL-1. It also exhibited reasonable specificity, resisting interference from three other pathogenic bacteria. These findings indicate that the proposed approach offers a cost-effective THz biosensing strategy that can be easily fabricated and conveniently operated to aid the diagnosis of infectious disease and food safety control.

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