RESUMO
An elevated level of homocysteine (Hcy) in serum is closely related to the development of various diseases. Therefore, homocysteine has been widely employed as a biomarker in medical diagnosis and the on-site detection of homocysteine is highly desired. In this study, a truncated highly specific aptamer for homocysteine was screened and used to design a lateral flow strip (LFS) for the detection of homocysteine. The aptamer was derived from a previously reported sequence. Based on the result of molecular docking, the original sequence was subjected to truncation, resulting in a reduction of the length from 66 nt to 55 nt. Based on the truncated aptamer, the LFS was designed for the detection of homocysteine. In the presence of homocysteine, the aptamer selectively binds to it, releasing cDNA from the aptamer/cDNA duplex. This allows cDNA to bind to the capture probe immobilized on the T zone of the strip, resulting in a red signal on the T zone from gold nanoparticles (AuNPs). The strip enables the visual detection of homocysteine in 5 min. Quantitative detection can be facilitated with the aid of ImageJ software. In this mode, the linear detection range for homocysteine is within 5-50 µM, with a detection limit of 4.18 µM. The strip has been effectively utilized for the detection of homocysteine in human serum. Consequently, the combination of the truncated aptamer and the strip offers a method that is sensitive, quick, and economical for the on-site detection of homocysteine.
Assuntos
Aptâmeros de Nucleotídeos , Ouro , Homocisteína , Nanopartículas Metálicas , Homocisteína/sangue , Homocisteína/química , Homocisteína/análise , Aptâmeros de Nucleotídeos/química , Humanos , Ouro/química , Nanopartículas Metálicas/química , Limite de Detecção , Técnicas Biossensoriais/métodos , Fitas Reagentes/química , Simulação de Acoplamento MolecularRESUMO
The point-of-care testing (POCT) of miRNA has significant application in medical diagnosis, yet presents challenges due to their characteristics of high homology, low abundance, and short length, which hinders the achievement of quick detection with high specificity and sensitivity. In this study, a lateral flow assay based on the CRISPR/Cas13a system and MnO2 nanozyme was developed for highly sensitive detection of microRNA-21 (miR-21). The CRISPR/Cas13a cleavage system exhibits the ability to recognize the specific oligonucleotide sequence, where two-base mismatches significantly impact the cleavage activity of the Cas13a. Upon binding of the target to crRNA, the cleavage activity of Cas13a is activated, resulting in the unlocking of the sequence and initiating strand displacement, thereby enabling signal amplification to produce a new sequence P1. When applying the reaction solution to the lateral flow test strip, P1 mediates the capture of MnO2 nanosheets (MnO2 NSs) on the T zone, which catalyzes the oxidation of the pre-immobilized colorless substrate 3,3',5,5'-tetramethylbenzidine (TMB) on the T zone and generates the blue-green product (ox-TMB). The change in gray value is directly proportional to the concentration of miR-21, allowing for qualitative detection through visual inspection and quantitative measurement using ImageJ software. This method achieves the detection of miR-21 within a rapid 10-min timeframe, and the limit of detection (LOD) is 0.33 pM. With the advantages of high specificity, simplicity, and sensitivity, the lateral flow test strip and the design strategy hold great potential for the early diagnosis of related diseases.
Assuntos
Técnicas Biossensoriais , Sistemas CRISPR-Cas , Compostos de Manganês , MicroRNAs , Nanoestruturas , Óxidos , Humanos , Técnicas Biossensoriais/métodos , Limite de Detecção , Compostos de Manganês/química , MicroRNAs/análise , Nanoestruturas/química , Óxidos/químicaRESUMO
To investigate the efficacy of laser acupuncture and photobiomodulation therapy in alleviating symptoms among patients diagnosed with Bell's palsy with duration of greater than 8 weeks. The randomized controlled trial has been performed from May 2021 to April 2023. Patients were eligible who had Bell's palsy with duration of greater than 8 weeks on out-patient Department of Otorhinolaryngology in Beijing Tongren Hospital. The laser acupuncture group received class IV laser treatment for 3 times per weeks, a total of 72 times. The control group received the same treatment procedure except the laser parameter. The primary outcome measures comprised House-Brackmann facial nerve grading system and electroneurography. Secondary outcome measures comprised Sunnybrook facial grading system, electromyography, and the blink reflex. A total of 84 participants were included (42 control group, 42 laser acupuncture group). After treatment, House-Brackmann facial nerve grading system (OR, 0.11; 95% CI, 0.04-0.30; P < 0.001), and the pathologic numbers of electroneuronography were statistically different between the laser acupuncture group and control group, including orbicularis oculi (OR,0.08; 95% CI, 0.02-0.21; P < 0.001), Frontalis muscle (OR,0.14; 95% CI, 0.05-0.39; P < 0.001), Orbicularis oris (OR,0.13; 95% CI, 0.04-0.36; P < 0.001), Ala nasi muscle (OR,0.06; 95% CI, 0.02-0.18; P < 0.001). In secondary outcomes, Sunnybrook facial grading system, has significant difference between the two groups (20.26; 95% CI, 14.69 to 25.83; P < 0.01). Latency by ENoG, include orbicularis oculi (-0.61; 95% CI, -0.43 to -0.09; P < 0.001), frontalis muscle (-0.12; 95% CI, -0.21 to -0.03; P < 0.01), orbicularis oris (-0.28; 95% CI, -0.41 to -0.16; P < 0.001), and ala nasi muscle (-0.26; 95% CI, -0.38 to -0.16; P < 0.001). All amplitudes of MUAPs and durations by electromyography (EMG) showed statistically significant differences compared with the control group after treatment. For the frontalis muscle, the amplitude of MUAPs was -64.23 (95% CI, -80.89 to -47.56; P < 0.001) and duration was -1.18 (95% CI, -1.49 to -0.87; P < 0.001). For orbicularis oris, amplitude of MUAPs was -29.82 (95% CI, -55.03 to -4.62; P = 0.02) and duration was -0.57 (95% CI, -0.94 to -0.20; P < 0.001). For depressor angulli oris, amplitude of MUAPs was -47.06 (95% CI, -62.15 to -31.97; P < 0.001) and duration was -2.21 (95% CI, -2.69 to -1.72; P < 0.001). Blink reflex, including R1 (OR, 0.03; 95% CI, 0.01-0.16; P < .001), R2 (OR, 0.04; 95% CI, 0.004-0.29; P < .001), and R2 latency differences (OR, 0.15; 95% CI, 0.05-0.51; P < .001), have significant difference between the two groups, respectively. The findings suggest that laser acupuncture relieve symptoms for patients with Bell's palsy with a duration of greater than 8 weeks.Trial registration: ClinicalTrials.gov Identifier: NCT05846217.
Assuntos
Terapia por Acupuntura , Paralisia de Bell , Terapia com Luz de Baixa Intensidade , Humanos , Paralisia de Bell/radioterapia , Nervo Facial , Terapia por Acupuntura/métodos , Eletromiografia/métodosRESUMO
BACKGROUND: Meniere Disease is a clinical condition defined by hearing loss, tinnitus, and aural fullness symptoms, there are currently no any medications approved for its treatment. OBJECTIVE: To determine whether taVNS as an adjunctive therapy could relieve symptoms and improve the quality of life in patients with Meniere disease. METHODS: In this Single-center, single blind, randomized trial, participants were assigned to transcutaneous auricular vagus nerve stimulation (taVNS) group and sham taVNS group. The primary outcome measures comprised Tinnitus Handicap Inventory, Dizziness Handicap Inventory, Pure Tone Auditory, Visual analogue scale of aural fullness. Secondary outcome measures comprised the 36-Item Short Form Health Survey, video head impulse test, and the caloric test. RESULTS: After 12 weeks, the THI (-11.00, 95%CI, -14.87 to -7.13; P < 0.001), DHI (-47.26, 95%CI, -50.23 to -44.29; P < 0.001), VAS of aural fullness (-2.22, 95%CI, -2.95 to -1.49; P<0.01), and Pure Tone Thresholds (-7.07, 95%CI, -9.07 to -5.06; P<0.001) were significantly differed between the two groups. In addition, SF36(14.72, 95%CI, 11.06 to 18.39; P < 0.001), vHIT (RD, 0.26, 95 % CI, -0.44 to -0.08, RR, 0.43, 95 % CI, 0.22 to 0.83, P < 0.01), and the caloric test (RD, -0.24, 95 % CI, -0.43 to -0.04, RR, 0.66, 95 % CI, 0.44 to 0.95, P = 0.02) have significant difference between two group, respectively. CONCLUSIONS: These findings suggest that taVNS combined with Betahistine Mesylate relieve symptoms and improve the quality of life for patients with Meniere Disease. taVNS can be considered an adjunctive therapy in treatment of Meniere Disease. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT05328895.
Assuntos
Doença de Meniere , Zumbido , Estimulação Elétrica Nervosa Transcutânea , Estimulação do Nervo Vago , Humanos , Doença de Meniere/terapia , beta-Histina/uso terapêutico , Método Simples-Cego , Qualidade de Vida , Nervo Vago/fisiologiaRESUMO
Objective: To determine whether photobiomodulation therapy (PBMT) by class IV Multiwave Locked System laser treatment as an adjunctive therapy could relieve symptoms in patients with Bell's palsy with a duration of greater than 8 weeks. Materials and methods: This nonrandomized controlled trial was conducted from January 2020 to December 2022. Patients were eligible if they had Bell's palsy with a duration of greater than 8 weeks at the out-patient department of otorhinolaryngology in Beijing Tongren Hospital. The control group consisted of patients recruited between January 1, 2020, and December 31, 2020. The PBMT group consisted of patients recruited between January 1, 2021, and December 31, 2022. In this study, the PBM used has a wavelength of 808 and 905 nm, 1.2 W power (808 nm is 1 W, 905 nm is 200 mW), continuous mode emission (808 nm) and pulsed mode emission (905 nm), 8.35 J/cm2 dosimetry, administered 3 times per week, 72 times of total treatment. The primary outcome measures included the House-Brackmann facial nerve grading system, the Sunnybrook facial grading system, and the Facial Clinimetric Evaluation Scale (FaCE). Secondary outcome measures comprised electroneurography, electromyography, and the blink reflex. Results: A total of 54 participants were included (27 in the control group and 27 in the photobiomodulation group). After 6 months, the House-Brackmann grading system [risk difference, -0.59, confidence interval (95% CI), -0.81 to -0.38, relative risk, 0.27, 95% CI, 0.13-0.56, p < 0.001], Sunnybrook facial grading system (21.14, 95% CI, 11.71-30.58; p < 0.001), and FaCE (-0.20, 95% CI, 0.41-0.02; p = 0.07) had significant difference between the two groups. Latency of ala nasi muscle (10.92, 95% CI, 5.58-16.27; p < 0.001) was not statistically significant after treatment compared with the control group; however, most of the electrophysiological examinations have significant difference between the two groups, respectively. Conclusions: The results of this study suggest that PBMT may relieve symptoms for patients with Bell's palsy with a duration of greater than 8 weeks. Trial Registration: ClinicalTrials.gov Identifier: NCT05585333.
Assuntos
Paralisia de Bell , Paralisia Facial , Terapia com Luz de Baixa Intensidade , Humanos , Paralisia de Bell/radioterapia , Fototerapia , Manejo da DorRESUMO
ABSTRACT: We report a case of pendular seesaw nystagmus caused by bitemporal hemianopia; the nystagmus disappeared while in darkness as previously described, but it also disappeared with monocular occlusion, which indicates the pivotal role of binocular vision in the pathogenesis.
Assuntos
Nistagmo Patológico , Hemianopsia , Humanos , Nistagmo Patológico/diagnóstico , Nistagmo Patológico/etiologia , Nistagmo Patológico/patologia , Visão BinocularRESUMO
BACKGROUND: Charcot-Marie-Tooth disease (CMT) is a genetically heterogeneous hereditary neuropathy, and CMT1A is the most common form; it is caused by a duplication of the peripheral myelin protein 22 (PMP22) gene. Mutations in the transient sodium channel Nav1.4 alpha subunit (SCN4A) gene underlie a diverse group of dominantly inherited nondystrophic myotonias that run the spectrum from subclinical myopathy to severe muscle stiffness, disabling weakness, or frank episodes of paralysis. CASE PRESENTATION: We describe a Chinese family affected by both CMT1A and myotonia with concomitant alterations in both the PMP22 and SCN4A genes. In this family, the affected proband inherited the disease from his father in an autosomal dominant manner. Genetic analysis confirmed duplication of the PMP22 gene and a missense c.3917G > C (p. Gly1306Ala) mutation in SCN4A in both the proband and his father. The clinical phenotype in the proband showed the combined involvement of skeletal muscle and peripheral nerves. Electromyography showed myopathic changes, including myotonic discharges. MRI revealed the concurrence of neurogenic and myogenic changes in the lower leg muscles. Sural nerve biopsies revealed a chronic demyelinating and remyelinating process with onion bulb formations in the proband. The proband's father presented with confirmed subclinical myopathy, very mild distal atrophy and proximal hypertrophy of the lower leg muscles, pes cavus, and areflexia. CONCLUSION: This study reports the coexistence of PMP22 duplication and SCN4A mutation. The presenting features in this family suggested that both neuropathy and myopathy were inherited in an autosomal dominant manner. The proband had a typical phenotype of sodium channel myotonia (SCM) and CMT1A. However, his father with the same mutations presented a much milder clinical phenotype. Our study might expand the genetic and phenotypic spectra of neuromuscular disorders with concomitant mutations.
Assuntos
Artrogripose , Doença de Charcot-Marie-Tooth , Miotonia , Doença de Charcot-Marie-Tooth/complicações , Doença de Charcot-Marie-Tooth/genética , Humanos , Masculino , Proteínas da Mielina , Canal de Sódio Disparado por Voltagem NAV1.4/genética , ProteínasRESUMO
The traditional gold nanoparticle (AuNP) growth-based plasmonic ELISA (pELISA) strictly and directly controlled by reducing reagents can achieve high sensitivity, but it remains fragile toward the surrounding environment. This work developed a sandwich pELISA for Cronobacter detection in powdered infant formula samples by mediating AuNP growth through DNA. In this assay, DNA adsorbed on the surface of gold nanoseeds guided the anisotropic crystal growth with hydroxylamine as a reducing reagent, and the catalase-hydrogen peroxide (Cat-H2O2) system was introduced to bridge the DNA-directed AuNP growth and pELISA, as such DNA can be cleaved into fragments by the hydroxyl radical generated from oxidation of H2O2 through Fenton reagents. Under optimized conditions, the proposed pELISA can qualitatively detect Cronobacter species (Cronobacter muytjensii ATCC 51329) by the naked eye with a cut-off limit of 3 × 105 cfu/mL. This method also revealed a good linear range (3 × 102 to 3 × 107 cfu/mL) for quantitative detection of C. muytjensii ATCC 51329 with a limit of detection of 1.6 × 102 cfu/mL, which is approximately 162.5 times lower than that of horseradish peroxidase-based conventional ELISA (2.6 × 104 cfu/mL). By taking advantage of highly stable DNA-directed AuNP growth, the proposed method shows a good performance in powdered infant formula samples spiked with different concentrations of C. muytjensii ATCC 51329 with average recoveries ranging from 90.79 to 119.09% and coefficient of variation ranging from 4.24 to 9.55%. These values corresponded to an acceptable accuracy and precision for the proposed method. In brief, this work shows potential for screening other analytes in food safety, clinical diagnostics, and environmental monitoring.
Assuntos
Cronobacter/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/métodos , Ouro , Fórmulas Infantis/microbiologia , Nanopartículas Metálicas , Cronobacter/genética , Cronobacter sakazakii/genética , DNA Bacteriano/isolamento & purificação , Microbiologia de Alimentos , Inocuidade dos Alimentos , Ouro/química , Humanos , Peróxido de Hidrogênio , Ferro , Nanopartículas Metálicas/química , PósRESUMO
Herein, we report a novel magnet-mediated antibody-boronate sandwich-typed assay (ABSTA) strategy for the ultrasensitive, specific, rapid, and enzyme-free detection of glycoproteins in complex samples. The proposed ABSTA method exhibited ultrahigh sensitivity for HCG with a detection limit of 0.19 mIU mL-1, which is approximately 40-fold lower than that of conventional sandwich enzyme immunoassay.
Assuntos
Anticorpos Imobilizados/química , Ácidos Borônicos/química , Gonadotropina Coriônica/sangue , Corantes Fluorescentes/química , Imãs/química , Anticorpos Monoclonais/química , Gonadotropina Coriônica/análise , Fluorescência , Humanos , Imunoensaio/métodos , Limite de DetecçãoRESUMO
A novel and ultrasensitive electrochemiluminescent (ECL) bioassay has been developed for the detection of atrazine (ATZ), whereby bifunctional S, N-codoped carbon dots (S, N-CDs) and activated mesoporous bicarbons (BCs) have been innovatively integrated to synergistically amplify the ECL signal. When endogenous dissolved O2 is used as a coreactant, its sluggish reduction hinders the enhancement of ECL intensity of the luminophore, thus restricting its further application in bioanalysis. Here, bifunctional S, N-CDs severe as not only the ECL luminophore but the coreaction accelerator of dissolved O2 to generate more intermediates to boost the coreaction without using any other coreactant and coreaction accelerator. The as-formed nanoarchitectures of BCs possess enlarged surface area as the nanocarriers. They could provide adequate active sites for immobilization of tyrosinase (Tyr), which greatly prompts the ECL bioassay applications. Such a smart integration of bifunctional S, N-CDs, activated mesoporous BCs and the enzyme inhibition reaction achieves a unique and attractive high-performance signal-on ECL bioassay, realizing ultrasensitive detection of ATZ. Under the optimal condition, this bioassay exhibits two linear ranges, from 0.0001 to 0.01⯵gâ¯L-1 and 0.01-20⯵gâ¯L-1 with a detection limit of 0.08â¯ngâ¯L-1â¯at signal to noise ratio of 3. The as-fabricated assay is sensitive and economical, opening a new way for the enhancement of ECL signal output and a versatile strategy for ultrasensitive ECL bioanalysis.
Assuntos
Atrazina/análise , Técnicas Biossensoriais , Técnicas Eletroquímicas , Medições Luminescentes , Monofenol Mono-Oxigenase/química , Nanopartículas/química , Atrazina/metabolismo , Carbono/química , Monofenol Mono-Oxigenase/metabolismo , Nanopartículas/metabolismo , Tamanho da Partícula , Porosidade , Pontos Quânticos/química , Propriedades de SuperfícieRESUMO
The rapid and sensitive detection of foodborne pathogens is one of the most important issues in food safety control. In this work, we developed a novel fluorescence immunoassay method for the sensitive detection of Salmonella choleraesuis. The method uses the fluorescent signals of histone-ds-poly(AT)-templated copper nanoparticles (His-pAT CuNP) as signal transducers and glucose oxidase as an alternative for horseradish peroxidase for the generation of hydrogen peroxide (H2O2) through the catalysis of glucose. The H2O2 is then further converted into hydroxyl radical (·OH) by Fenton reagents. Owing to the ultrahigh sensitivity of His-pAT CuNP synthesis toward ·OH, the proposed fluorescence immunoassay method exhibited excellent sensitivity for S. choleraesuis, with a limit of detection of 8.04 × 101 cfu/mL, which is 3 orders of magnitude lower than that of the tetramethylbenzidine-based traditional immunoassay. The reliability of the proposed method was evaluated by using spiked milk samples with S. choleraesuis concentration ranging from 8.8 × 101 to 8.8 × 104 cfu/mL. The average recoveries for the intra- and inter-assay ranged from 73.52 to 96.59% and from 66.99 to 98.24% with a coefficient of variation from 6.85 to 31.26% and 5.46 to 17.99%, respectively. These results indicated that the proposed fluorescence immunoassay possesses a great potential for ultra-sensitive detection of foodborne pathogens in food safety control.
Assuntos
Fluorimunoensaio/veterinária , Histonas/química , Nanopartículas Metálicas , Leite/microbiologia , Salmonella enterica/isolamento & purificação , Animais , Bovinos , Cobre , Fluorimunoensaio/métodos , Glucose Oxidase/metabolismo , Peróxido de Hidrogênio/química , Ferro , Nanopartículas Metálicas/química , Reprodutibilidade dos TestesRESUMO
We applied urease-induced silver metallization on the surface of gold nanorods (AuNR) to improve colorimetric ELISA for the rapid and sensitive detection of Salmonella enterica Choleraesuis. To this end, we introduced a biotin-streptavidin system as a bridge to determine the correlation between urease and S. enterica Choleraesuis concentrations. The captured urease can catalyze the hydrolysis of urea into carbon dioxide and ammonia, and the generated ammonia can then induce the deposition of silver shell on the surface of AuNR in the presence of silver nitrate and glucose. With the decreased aspect ratio (length divided by width) of AuNR, a multicolor change of AuNR solution and a significant blue shift in the longitudinal localized surface plasmon resonance absorption peak (Δλmax) of AuNR were obtained at the target concentrations of 1.21 × 101 to 1.21 × 108 cfu/mL. Consequently, the detection limits of our proposed colorimetric ELISA were as low as 1.21 × 102 cfu/mL for qualitative detection with naked eyes, and 1.21 × 101 cfu/mL for quantitative detection, in which changes in Δλmax of AuNR were recorded with a microplate reader. These values were at least 2 to 3 orders of magnitude lower than those obtained with conventional horseradish peroxidase-based ELISA. The analytical performance of our developed colorimetric ELISA in terms of selectivity, accuracy, reliability, and practicability were investigated by analyzing S. enterica Choleraesuis-spiked pasteurized whole milk samples.
Assuntos
Colorimetria/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Ouro/química , Leite/microbiologia , Nanotubos/química , Salmonella enterica/isolamento & purificação , Urease/química , Animais , Colorimetria/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Microbiologia de Alimentos/métodosRESUMO
Herein, for the first time we report a novel competitive fluorescence immunoassay for the ultrasensitive detection of aflatoxin B1 (AFB1) using histone-ds-poly(AT) templated copper nanoparticles (His-pAT CuNPs) as the fluorescent indicator. In this immunoassay, glucose oxidase (Gox) was used as the carrier of the competing antigen to catalyze the formation of hydrogen peroxide (H2O2) from glucose. H2O2 was converted to a hydroxyl radical using Fenton's reagent, which further regulated the fluorescence signals of His-pAT CuNPs. Owing to the ultrahigh sensitivity of the ss-DNA to the hydroxyl radical, the proposed fluorescence immunoassay exhibited a favorable dynamic linear detection of AFB1 ranging from 0.46 pg mL-1 to 400 pg mL-1 with an good half maximal inhibitory concentration and limit of detection of 6.13 and 0.15 pg mL-1, respectively. The intra- and inter-assay showed that the average recoveries for AFB1 spiked corn samples ranged from 96.87% to 100.73% and 96.67% to 114.92%, respectively. The reliability of this method was further confirmed by adopting ultra-performance liquid chromatography coupled with the fluorescence detector method. In summary, this work offers a novel screening strategy with high sensitivity and robustness for the quantitative detection of mycotoxins or other pollutants for food safety and clinical diagnosis.
Assuntos
Aflatoxina B1/análise , Cobre/química , DNA de Cadeia Simples/metabolismo , Imunoensaio , Nanopartículas Metálicas/química , Espectrometria de Fluorescência , DNA de Cadeia Simples/química , Glucose Oxidase/metabolismo , Histonas/química , Peróxido de Hidrogênio/química , Ferro/química , Poli A/química , Poli T/química , Reprodutibilidade dos TestesRESUMO
Herein, we present a novel sandwich fluorescence enzyme linked immunosorbent assay (ELISA) for highly sensitive detection of Hepatitis B virus surface antigen (HBsAg) based on glucose oxidase (GOx)-induced fluorescence quenching of mercaptopropionic acid-modified CdTe quantum dots (MPA-QDs). In this system, hydrogen peroxide (H2O2) sensitive MPA-QDs was used as a signal output, and glucose oxidase (GOx) was used as label which can generate H2O2 via catalytic oxidation of glucose. The proposed method showed dynamic linear detection of HBsAg both in the range of 47pgmL-1 ~ 380pgmL-1 and 0.75ngmL-1 ~ 12.12ngmL-1. The detection limit of the proposed fluorescence ELISA was 1.16pgmL-1, which was approximately 430-fold lower than that of horseradish peroxidase (HRP)-based conventional ELISA. The average recoveries for HBsAg-spiked serum samples ranged from 98.0% to 126.8% with the relative standard derivation below 10%, thus indicating acceptable precision and high reproducibility of the proposed fluorescence ELISA for HBsAg detection. Additionally, the developed method showed no false positive results analyzing 35 real HBsAg-negative serum samples, and exhibited excellent agreement (R2=0.9907) with a commercial time-resolved fluorescence immunoassay (TRFIA) kit for detecting 31 HBsAg-positive serum samples. In summary, the proposed method based on fluorescence quenching of H2O2 sensitive QDs is considerably to be an excellent biodetection platform with ultrahigh sensitivity, good accuracy and excellent reliability.
Assuntos
Ensaio de Imunoadsorção Enzimática/métodos , Fluorescência , Glucose Oxidase/química , Hepatite B/diagnóstico , Pontos Quânticos/química , Fluorimunoensaio/métodos , Glucose/química , Glucose/metabolismo , Glucose Oxidase/metabolismo , Hepatite B/sangue , Hepatite B/virologia , Antígenos de Superfície da Hepatite B/análise , Antígenos de Superfície da Hepatite B/química , Antígenos de Superfície da Hepatite B/imunologia , Vírus da Hepatite B/imunologia , Vírus da Hepatite B/fisiologia , Peroxidase do Rábano Silvestre/química , Peroxidase do Rábano Silvestre/metabolismo , Humanos , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/metabolismo , Limite de Detecção , OxirreduçãoRESUMO
Efficient risk communication is a vital way to reduce the vulnerability of individuals when facing emergency risks, especially regarding earthquakes. Efficient risk communication aims at improving the supply of risk information and fulfilling the need for risk information by individuals. Therefore, an investigation into individual-level information seeking behavior within earthquake risk contexts is very important for improved earthquake risk communication. However, at present there are very few studies that have explored the behavior of individuals seeking earthquake risk information. Under the guidance of the Risk Information Seeking and Processing model as well as relevant practical findings using the structural equation model, this study attempts to explore the main determinants of an individual's earthquake risk information seeking behavior, and to validate the mediator effect of information need during the seeking process. A questionnaire-based survey of 918 valid respondents in Songyuan, China, who had been hit by a small earthquake swarm, was used to provide practical evidence for this study. Results indicated that information need played a noteworthy role in the earthquake risk information seeking process, and was detected both as an immediate predictor and as a mediator. Informational subjective norms drive the seeking behavior on earthquake risk information through both direct and indirect approaches. Perceived information gathering capacity, negative affective responses and risk perception have an indirect effect on earthquake risk information seeking behavior via information need. The implications for theory and practice regarding risk communication are discussed and concluded.
Assuntos
Terremotos , Comportamento de Busca de Informação , Adulto , China , Feminino , Humanos , Masculino , Percepção , RiscoRESUMO
Along with the progressive acceleration of urbanization, the need to identify potentially troublesome "Not In My Back Yard" (NIMBY) facilities in the city is inevitable. To resolve NIMBY conflict, it is important to know people's NIMBY risk acceptability for these facilities. A questionnaire survey was used among Chinese and Japanese college students to identify NIMBY risk acceptability. LISREL was used to construct a structural equation model to analyze the difference in NIMBY risk acceptability between the Chinese and Japanese college students. Factors that may affect NIMBY risk acceptability were analyzed: "perceiving utility," "perceiving risk," "trust in government," "reasonable compensation," and "procedural justice." The findings show that Japanese students' concerns were greater than Chinese students' concerns. Perceiving utility and perceiving risk were the most important factors that affect people's NIMBY risk acceptability, followed by procedural justice, trust in government, and reasonable compensation. There is a difference between the different cultural backgrounds in confronting the risk: Chinese students focus more on the reputation and value of real estate, while Japanese students pay more attention to environmental pollution and damage to health. Furthermore, cultural influences play a role in students' risk perception. To improve the risk acceptability for NIMBY facilities and provide a basis for resolving NIMBY conflicts, it is necessary to ensure the benefits of the NIMBY facility while reducing environmental pollution. The findings of this study may be of interest for policy makers and practitioners to devise future NIMBY strategies.
Assuntos
Opinião Pública , Risco , Estudantes/psicologia , Adulto , China , Cidades , Monitoramento Ambiental , Poluição Ambiental/estatística & dados numéricos , Feminino , Habitação , Humanos , Japão , Masculino , Estudantes/estatística & dados numéricos , Inquéritos e Questionários , Adulto JovemRESUMO
alpha-Zearalanol (alpha-ZAL), a phytochemical with both antioxidant and estrogen-like properties, has been shown to retard progression of atherosclerosis and regulate cardiovascular function in part through suppression of endothelin-1 (ET-1) secretion. However, the precise nature behind alpha-ZAL-elicited inhibition on ET-1 cascade is not largely known. Oxidized low density lipoprotein (oxLDL) plays a critical role in the expression and secretion of ET-1 as well as the onset and progression of atherosclerosis through accumulation of reactive oxygen species (ROS) and activation of mitogen-activated protein kinase stress signaling cascade. Therefore, this study was designed to examine the effect of alpha-ZAL on oxLDL-induced extracellular signal-regulated kinase (ERK) phosphorylation, ROS generation, activation of the transcriptional factor activator protein-1 (AP-1), expression, secretion and promoter activity of ET-1 in human umbilical vein endothelial cells (HUVEC). ROS generation was monitored using 2,7-dichlorofluorescin fluorescence. ET-1 expression and promoter activity were evaluated by RT-PCR and luciferase assays, respectively. oxLDL (35 microg/ml) significantly enhanced ERK phosphorylation, ROS generation, AP-1 activity, mRNA expression, secretion and promoter activity of ET-1 in HUVECs, all of which were abrogated by alpha-ZAL and the antioxidant N-acetyl-l-cysteine. Collectively, these data favor the notion that alpha-ZAL antagonizes oxLDL-induced upregulation of ET-1 gene expression and secretion via suppression of oxLDL-induced ROS accumulation, ERK phosphorylation, and activation of the endothelial transcriptional factor AP-1.
Assuntos
Endotelina-1/genética , Endotélio Vascular/efeitos dos fármacos , Lipoproteínas LDL/farmacologia , Zeranol/análogos & derivados , Células Cultivadas , Combinação de Medicamentos , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Humanos , Oxirredução , Fosforilação Oxidativa/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição AP-1/metabolismo , Veias Umbilicais/citologia , Veias Umbilicais/metabolismo , Zeranol/farmacologiaRESUMO
BACKGROUND: Oxidized low-density lipoprotein (oxLDL) promotes expression and secretion of endothelin-1 (ET-1), however, the precise mechanism involved is unclear. This study was designed to identify the regulatory mechanism of oxLDL-induced ET-1 expression in endothelial cells. METHODS: ET-1 mRNA expression, secretion and promoter activity were evaluated by reverse transcriptase-PCR (RT-PCR), enzyme immunometric and luciferase assays, respectively. RESULTS: oxLDL (35 microg/ml) significantly enhanced reactive oxygen species (ROS), mRNA expression, secretion and promoter activity of ET-1 in human umbilical vein endothelial cells (HUVECs), all of which were nullified by the antioxidant N-acetyl cysteine (NAC). oxLDL stimulated the extracellular signal-regulated kinase (ERK) phosphorylation in HUVECs, which was blocked by NAC and the mitogen-activated protein/extracellular signal-regulated kinase (MEK) inhibitor PD98059. NAC and PD98059 stopped oxLDL-elicited increase in mRNA expression, secretion and promoter activity of ET-1. Fusion plasmids with decreasing length of 5'-flanking sequence of ET-1 from -566 bpLuc to -250 bpLuc displayed increased luciferase activity after 24 h of oxLDL treatment. Interestingly, fusion plasmid from -233 and -185 bpLuc significantly reduced the luciferase activity in control and oxLDL-treated HUVECs. In addition, transfection of the reporter construct -250Luc, which contains a 2 bp mutation at activator protein-1 site, abolished both basal and oxLDL-stimulated ET-1 promoter activities. CONCLUSION: Collectively, our data favor the notion that oxLDL stimulates ERK phosphorylation via ROS accumulation, which in turn stimulates vascular endothelial transcriptional factor activator protein-1 and ET-1 expression as well as secretion.
Assuntos
Células Endoteliais/metabolismo , Endotelina-1/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/fisiologia , Lipoproteínas LDL/fisiologia , Espécies Reativas de Oxigênio , Células Cultivadas , Endotelina-1/genética , Regulação da Expressão Gênica , Humanos , Fosforilação , Transdução de Sinais , Veias Umbilicais/metabolismoRESUMO
This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor-in-Chief. After an institutional investigation into the work of Dr. Jun Ren, University of Wyoming subsequently conducted an examination of other selected publications of Dr. Ren's under the direction of the HHS Office of Research Integrity. Based on the findings of this examination, the University of Wyoming recommended this article be retracted due to data irregularities in Figures 3 and 5 that significantly affect the results and conclusions reported in the manuscript.
Assuntos
Células Endoteliais/fisiologia , Endotelina-1/metabolismo , Homocisteína/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Fitoestrógenos/farmacologia , Zeranol/análogos & derivados , Células Cultivadas , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas Quinases Ativadas por Mitógeno/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição AP-1/efeitos dos fármacos , Ativação Transcricional , Veias Umbilicais , Regulação para Cima , Zeranol/farmacologiaRESUMO
Oxidative modification of low-density lipoprotein (LDL) leads to formation of the atherogenic molecule oxidized LDL (oxLDL), which is considered to be an important mediator for vascular endothelial dysfunction and atherosclerosis. It is speculated that reduced nitric oxide (NO) release/bioavailability and enhanced release of endothelin-1 (ET-1) may contribute to oxLDL-induced endothelial dysfunction. Estrogen may improve lipid profile and inhibit oxLDL-induced endothelial damage. However, estrogen replacement therapy has been suspended due to uncertainty in benefits versus risk (such as cancer progression) in postmenopausal women. This study was designed to evaluate the effect of a novel phytoestrogen, alpha-zearalanol (alpha-ZAL), on oxLDL-induced effect on NO and ET-1 production in human umbilical vein endothelial cells (HUVEC). HUVEC were incubated with oxLDL (50 microg/mL) for 24 h in the absence or presence of alpha-ZAL (0-1000 nM), 17beta-estradiol (E2, 10 nM), or the E2 receptor antagonist ICI182780 (1 microM). Levels of NO and ET-1 were measured by spectrophotometry and enzymatic immunoassay, respectively. NOS activity was evaluated by conversion of 3H-arginine to 3H-citrulline. Protein and mRNA expression of NOS and ET-1 were measured by Western blot and RT-PCR. Our results indicated that oxLDL significantly reduced NO release and NOS activity, and enhanced ET-1 pro-duction associated with reduced NOS3 (but not NOS2) expression and enhanced ET-1 mRNA expression. All these oxLDL-induced alterations were significantly attenuated or abolished by co-incubation with alpha-ZAL or E2, both through an E2 receptor-dependent mechanism. alpha-ZAL, E2, and ICI182780 had no effect on NO/ET-1 release, NOS activity, or expression of NOS and ET-1. These data suggested that the phytoestrogen alpha-ZAL, like E2, may effectively antagonize oxLDL-induced decrease in NO and increase in ET-1, which may be protective for endothelial function.