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1.
Opt Lett ; 49(13): 3806-3809, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38950276

RESUMO

Current non-confocal non-line-of-sight (NLOS) imaging faces the problems of low resolution and limited scene adaptability. We propose a non-confocal NLOS imaging method based on spherical-slice transform from spatial and temporal frequency to space and time. Simulation and experimental results show that the proposed method has high-resolution reconstruction without artifact interference, shape distortion, and position offset. Furthermore, it has strong scene adaptability. After GPU acceleration, the reconstruction time of the proposed method can be reduced to several hundred milliseconds for the PF32 photon array camera with 32 × 32 detection units. In the future, the proposed method has great potential for application in real-time NLOS imaging systems.

2.
J Environ Manage ; 365: 121613, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38944964

RESUMO

Composting is a biological reaction caused by microorganisms. Composting efficiency can be adequately increased by adding biochar and/or by inoculating with exogenous microorganisms. In this study, we looked at four methods for dewatered sludge waste (DSW) and wheat straw (WS) aerobic co-composting: T1 (no additive), T2 (5% biochar), T3 (5% of a newly isolated strain, Xenophilus azovorans (XPA)), and T4 (5% of biochar-immobilized XPA (BCI-XPA)). Throughout the course of the 42-day composting period, we looked into the carbon dynamics, humification, microbial community succession, and modifications to the driving pathways. Compared to T1 and T2, the addition of XPA (T3) and BCI-XPA (T4) extended the thermophilic phase of composting without negatively affecting compost maturation. Notably, T4 exhibited a higher seed germination index (132.14%). Different from T1 and T2 treatments, T3 and T4 treatments increased CO2 and CH4 emissions in the composting process, in which the cumulative CO2 emissions increased by 18.61-47.16%, and T3 and T4 treatments also promoted the formation of humic acid. Moreover, T4 treatment with BCI-XPA addition showed relatively higher activities of urease, polyphenol oxidase, and laccase, as well as a higher diversity of microorganisms compared to other processes. The Functional Annotation of Prokaryotic Taxa (FAPROTAX) analysis showed that microorganisms involved in the carbon cycle dominated the entire composting process in all treatments, with chemoheterotrophy and aerobic chemoheterotrophy being the main pathways of organic materials degradation. Moreover, the presence of XPA accelerated the breakdown of organic materials by catabolism of aromatic compounds and intracellular parasite pathways. On the other hand, the xylanolysis pathway was aided in the conversion of organic materials to dissolved organics by the addition of BCI-XPA. These findings indicate that XPA and BCI-XPA have potential as additives to improve the efficiency of dewatered sludge and wheat straw co-composting.


Assuntos
Carbono , Compostagem , Esgotos , Triticum , Esgotos/microbiologia , Carbono/metabolismo , Substâncias Húmicas , Carvão Vegetal
3.
Anal Chem ; 95(20): 8097-8104, 2023 05 23.
Artigo em Inglês | MEDLINE | ID: mdl-37171156

RESUMO

Long noncoding RNAs (lncRNAs) play pivotal roles in multifarious physiological and pathological processes, and their aberrant expression may disturb the normal regulatory network of gene expression to induce diverse human diseases. Herein, we construct a fluorescent light-up biosensor with a low background for label-free detection of lncRNAs by coupling duplex-specific nuclease (DSN)-assisted target recycling amplification with transcription-driven synthesis of fluorogenic RNA aptamer-Corns. We design two linear probes, including a capture probe for initiating a cyclic cleavage reaction and a linear template for transcribing RNA aptamer-Corn. Target lncRNA is recognized by capture probes assembled on magnetic bead (MB) surfaces to trigger a DSN-assisted cyclic cleavage reaction, releasing abundant T7 promoter sequences. After magnetic separation, free T7 promoter hybridizes with a linear template to induce efficient transcription amplification with the assistance of T7 RNA polymerase, producing numerous fluorogenic RNA aptamer-Corns that can light up small-molecule fluorogens 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO). Notably, the introduction of MBs facilitates both the separation of cleaved capture probes and the enrichment/isolation of target lncRNAs from the complex biological matrix. Benefiting from the high efficiency of DSN/T7 RNA polymerase-mediated cascade amplification and high signal-to-background ratio of the Corn-DFHO complex, this biosensor is capable of sensitively quantifying lncRNA with a detection limit of 31.98 aM. Moreover, it can precisely quantify lncRNA at the single-cell level and even in complex biological samples, and it can differentiate tumor cells from normal cells. Importantly, this Corn-based biosensor is readily extended to detect other lncRNAs by altering capture probe sequences, opening a new avenue for molecular diagnosis.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Calosidades , RNA Longo não Codificante , Humanos , RNA Longo não Codificante/genética , Zea mays/genética , Corantes , Limite de Detecção
4.
Anal Chem ; 95(5): 3082-3088, 2023 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-36692970

RESUMO

Long noncoding RNAs (lncRNAs) are valuable biomarkers and therapeutic targets, and they play essential roles in various pathological and biological processes. So far, the reported lncRNA assays usually suffer from unsatisfactory sensitivity and time-consuming procedures. Herein, we develop a mix-and-read assay based on multiple cyclic enzymatic repairing amplification (ERA) for sensitive and rapid detection of mammalian metastasis-associated lung adenocarcinoma transcript 1 (lncRNA MALAT1). In this assay, we design two three-way junction (3WJ) probes including a 3WJ template and a 3WJ primer to specifically recognize lncRNA MALAT1, and the formation of a stable 3WJ structure induces cyclic ERA to generate triggers. The resulting triggers subsequently hybridize with a free 3WJ template and act as primers to initiate new rounds of cyclic ERA, generating abundant triggers. The hybridization of triggers with signal probes forms stable double-stranded DNA duplexes that can be specifically cleaved by apurinic/apyrimidinic endonuclease 1 to produce a high fluorescence signal. This assay can be carried out in a mix-and-read manner within 10 min under an isothermal condition (50 °C), which is the rapidest and simplest method reported so far for the lncRNA MALAT1 assay. This method can sensitively detect lncRNA MALAT1 with a limit of detection of 0.87 aM, and it can accurately measure endogenous lncRNA MALAT1 at the single-cell level. Moreover, this method can distinguish lncRNA MALAT1 expression in breast cancer patient tissues and their corresponding healthy adjacent tissues. Importantly, the extension of this assay to different RNAs detection can be achieved by simply replacing the corresponding target recognition sequences.


Assuntos
Neoplasias , RNA Longo não Codificante , Humanos , DNA/química , RNA Longo não Codificante/genética
5.
Anal Chem ; 95(31): 11815-11822, 2023 08 08.
Artigo em Inglês | MEDLINE | ID: mdl-37489894

RESUMO

Poly(ADP-ribose) polymerase 1 (PARP-1) is responsible for catalyzing the creation of poly(ADP-ribose) polymer and involved in DNA replication and repair. Sensitive measurement of PARP-1 is critical for clinical diagnosis. However, the conventional electrostatic attraction-based PAPR-1 assays usually involve laborious procedures, poor sensitivity, and false positives. Herein, we demonstrate the construction of a dendritic nanoassembly-based fluorescent biosensor for electrostatic interaction-independent and label-free measurement of human PARP-1 in lung tumor tissues. When PARP-1 is present, the specific double-stranded DNA (dsDNA)-activated PARP-1 transfers the ADP-ribosyl group from nicotinamide adenine dinucleotide (NAD+)/biotinylated NAD+ to the PARP-1 itself, resulting in the formation of biotinylated dsDNA-PARP-1-PAR polymer bioconjugates that can be captured by magnetic beads. Upon the addition of TdT, APE1, and NH2-modified T-rich probe, the captured dsDNAs with dual 3'-OH termini initiate TdT-activated APE1-mediated hyperbranched amplification to produce abundant dendritic DNA nanoassemblies that can be stained by SYBR Green I to generate a high fluorescence signal. This biosensor is characterized by a template-free, electrostatic interaction-independent, high sensitivity, and label-free assay. It enables rapid (less than 3 h) measurement of PARP-1 with a limit of detection of 4.37 × 10-8 U/µL and accurate measurement of cellular PARP-1 activity with single-cell sensitivity. Moreover, it is capable of screening potential inhibitors and discriminating the PARP-1 level in normal person tissues and lung cancer patient tissues, with great potential in PARP-1-related clinical diagnosis and drug discovery.


Assuntos
Técnicas Biossensoriais , Inibidores de Poli(ADP-Ribose) Polimerases , Humanos , Inibidores de Poli(ADP-Ribose) Polimerases/farmacologia , NAD , Eletricidade Estática , Pulmão
6.
Anal Chem ; 95(40): 15133-15139, 2023 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-37751602

RESUMO

An abnormal expression level of long noncoding RNAs (lncRNAs) is implicated in multiple cancers, and their sensitive and rapid measurement is pivotal for early cancer diagnosis and cancer treatment. The conventional lncRNA assays often suffer from labor-intensive/time-consuming procedures and limited sensitivity. Herein, we report a simple and sensitive fluorescent biosensor for rapid and label-free measurement of lncRNAs based on recombinase polymerase amplification (RPA) without the involvement of thermal cycling and reverse transcription. Target lncRNAs can bind with the 5'-end of the DNA template to create a DNA-lncRNA hybrid, protecting the DNA template from RecJf exonuclease-mediated degradation. Subsequently, the primers hybridize with the intact DNA templates and are extended to generate the dsDNA products with the assistance of polymerase. The resultant dsDNA products may be amplified by exponential recombinase polymerase amplification to produce abundant dsDNAs, generating a distinct fluorescence signal within 10 min. This biosensor achieves a wide dynamic range from 10-17 to 10-9 M and high sensitivity with a detection limit of 1.23 aM. Moreover, it can distinguish the expressions of lncRNA HOTAIR in the tissues of healthy individuals and breast cancer patients, with broad application prospects in lncRNA-related research and early diagnosis of cancers.


Assuntos
Neoplasias da Mama , RNA Longo não Codificante , Humanos , Feminino , Recombinases , Exonucleases , RNA Longo não Codificante/genética , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/genética , Nucleotidiltransferases , DNA , Técnicas de Amplificação de Ácido Nucleico/métodos
7.
Anal Chem ; 95(35): 13201-13210, 2023 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-37603851

RESUMO

N6-Methyladenosine (m6A) demethylases can catalyze the removal of the methyl modification on m6A, and it is closely associated with the occurrence, proliferation, differentiation, and metastasis of malignancies. The m6A demethylases (e.g., fat mass and obesity-associated protein (FTO)) may act as a cancer biomarker and are crucial for anticancer drug screening and early clinical diagnosis. Herein, we demonstrate the construction of a quantum-dot-based Förster resonance energy-transfer (FRET) nanosensor through direct encoding of streptavidin-binding RNA aptamers (SA aptamers) for m6A demethylase detection. This nanosensor employs multiple Cy5-molecule-labeled SA aptamers as the building materials to construct the 605QD-RNA-Cy5 nanoassembly, and it exploits the hinder effect of m6A upon elongation and ligation reactions to distinguish m6A-containing RNA probes from demethylated RNA probes. When m6A demethylase is present, the m6A-containing RNA probes are demethylated to generate the demethylated RNA probes, initiating strand extension and ligation reactions to yield a complete transcription template for SA aptamers. Subsequently, a T7-assisted cascade transcription amplification reaction is activated to transcribe abundant SA aptamers with the incorporation of multiple Cy5 fluorophores. The Cy5-incorporated SA aptamers can self-assembly onto the streptavidin-coated 605QD surface to obtain the 605QD-SA aptamer-Cy5 nanoassemblies, resulting in the generation of distinct FRET signals. This nanosensor exhibits ultrahigh sensitivity and excellent specificity, and it can detect endogenous FTO at the single-cell level. Furthermore, this nanosensor can precisely measure enzyme kinetic parameters, screen m6A demethylase inhibitors, and differentiate the FTO expression between breast cancer patients and healthy individual tissues, offering a versatile platform for clinical diagnostic and drug discovery.


Assuntos
Aptâmeros de Nucleotídeos , Humanos , Estreptavidina , Transferência Ressonante de Energia de Fluorescência , Sondas RNA , Adenosina , Dioxigenase FTO Dependente de alfa-Cetoglutarato/genética
8.
Anal Chem ; 94(32): 11425-11432, 2022 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-35916620

RESUMO

N6-methyladenosine modification as an mRNA modification in mammalian cells is dynamically reversible, regulated by RNA demethylase [e.g., fat mass and obesity-associated protein (FTO)]. The abnormal expression of FTO is closely related to numerous diseases (e.g., various cancers and obesity). Herein, we demonstrate the single-molecule counting of FTO in human cancer cells and breast tissues based on a T7 RNA polymerase-mediated rolling circle transcription (RCT) amplification-driven clustered regularly interspaced short palindromic repeat (CRISPR)─Cas12a. When FTO is present, it demethylates the DNA substrate, initiating the DpnII-mediated cleavage reaction. After magnetic separation, the cleaved DNA fragments trigger the T7 RNA polymerase-mediated RCT amplification, activating CRISPR-/Cas12a-mediated cleavage of signal probes and releasing abundant FAM molecules that are simply counted via single-molecule detection. In this assay, only target FTO can generate CRISPR RNAs, efficiently improving detection specificity. Moreover, the integration of single-molecule detection with magnetic separation achieves zero background and effectively enhances detection sensitivity. This method can specifically and sensitively monitor FTO activity with a limit of detection of 1.20 × 10-13 M, and it may measure FTO at the single-cell level. Furthermore, it may accurately discriminate the FTO expression level in breast tissues between healthy persons and breast cancer patients and screen the FTO inhibitors as well, with great potential in clinical diagnosis and drug discovery.


Assuntos
Sistemas CRISPR-Cas , Neoplasias , Adenosina/metabolismo , Dioxigenase FTO Dependente de alfa-Cetoglutarato/genética , Dioxigenase FTO Dependente de alfa-Cetoglutarato/metabolismo , Animais , Humanos , Mamíferos/metabolismo , Neoplasias/genética , Obesidade/genética
9.
Anal Chem ; 94(4): 2119-2125, 2022 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-35050578

RESUMO

8-Oxoguanine DNA glycosylase is essential for maintaining genomic integrity and stability, while its abnormal activity may lead to the disturbance in the normal DNA damage repair and the occurrence of carcinogenicity and teratogenicity. Herein, we construct a CRISPR-Cas-based biosensor for rapid and sensitive measurement of 8-oxoguanine DNA glycosylases. This biosensor involves a hairpin probe and integrates quadratic strand displacement amplification (SDA) with a CRISPR/Cas12a effector with the characteristics of rapidity (within 40 min) and isothermal assay. The presence of 8-oxoguanine DNA glycosylase can initiate the quadratic SDA to produce large amounts of activators with the assistance of polynucleotide kinase (PNK). Subsequently, the activators can bind with crRNA to activate Cas12a, cleaving signal probes and recovering Cy5 fluorescence, which can be accurately quantified by single-molecule imaging. Notably, the designed hairpin probes can effectively block the hybridization of the generated activators with free hairpin probes, endowing this biosensor with high sensitivity. In addition, the utilization of PNK instead of apurinic/apyrimidinic endonuclease (APE1) greatly simplifies the experimental procedure to only a one-step reaction. The introduction of a single-molecule detection further reduces the sample consumption and improves the sensitivity. This biosensor displays a detection limit of 4.24 × 10-9 U µL-1, and it can accurately quantify cellular human 8-oxoguanine DNA glycosylase at a single-cell level. Furthermore, this biosensor can be applied for the screening of inhibitors, the analysis of kinetic parameters, and the discrimination of cancer cells from normal cells, with potential applications in molecular diagnostic and point-of-care testing.


Assuntos
Técnicas Biossensoriais , DNA Glicosilases , Sistemas CRISPR-Cas/genética , DNA Glicosilases/genética , DNA Glicosilases/metabolismo , Reparo do DNA , Guanina/análogos & derivados , Humanos
10.
Small ; 18(43): e2107048, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35229459

RESUMO

Single-crystalline cathodes are the most promising candidates for high-energy-density lithium-ion batteries (LIBs). Compared to their polycrystalline counterparts, single-crystalline cathodes have advantages over liquid-electrolyte-based LIBs in terms of cycle life, structural stability, thermal stability, safety, and storage but also have a potential application in solid-state LIBs. In this review, the development history and recent progress of single-crystalline cathodes are reviewed, focusing on properties, synthesis, challenges, solutions, and characterization. Synthesis of single-crystalline cathodes usually involves preparing precursors and subsequent calcination, which are summarized in the details. In the following sections, the development issues of single-crystalline cathodes, including kinetic limitations, interfacial side reactions, safety issues, reversible planar gliding and micro-cracking, and particle size distribution and agglomeration, are systematically analyzed, followed by current solutions and characterization techniques. Finally, this review is concluded with proposed research thrusts for the future development of single-crystalline cathodes.

11.
Nano Lett ; 21(10): 4193-4201, 2021 05 26.
Artigo em Inglês | MEDLINE | ID: mdl-33949866

RESUMO

Aberrant change in long noncoding RNA (lncRNA) is associated with various diseases and cancers. So far, simultaneous detection of lncRNAs has remained a great challenge due to their large size and extensive secondary structure. Herein, we develop an enzyme-free single-molecule/particle detection method for simultaneous detection of multiple lncRNAs in cancer cells based on target-catalyzed strand displacement. We designed the magnetic bead-capture probe-multiple Cy5/Cy3-modified reporter unit complexes to isolate and identify lncRNA MALAT1 and lncRNA HOTAIR. The target-catalyzed strand displacement reactions lead to the release of Cy5 and Cy3 fluorescent molecules from the complexes, which can be subsequently quantified by single-molecule/particle detection. The dual-targetability, good selectivity and high sensitivity of this method enables simultaneous detection of multiple lncRNAs in even single cancer cell. Importantly, this method can discriminate cancer cells from normal cells and has significant advantages in the simple sequence design and in being free of enzymes, holding great potential in living cell imaging and early clinical diagnosis.


Assuntos
Neoplasias , RNA Longo não Codificante , Neoplasias/diagnóstico , Neoplasias/genética , RNA Longo não Codificante/genética
12.
Phys Chem Chem Phys ; 22(6): 3604-3610, 2020 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-31995069

RESUMO

Plasmonic chirality has attracted more and more attention recently due to the enhanced chiroptical response and its potential applications in biosensing. Plasmonic Fano resonance arises from the interference between a dark narrow resonance and a bright broad resonance, and it provides a new paradigm to control the plasmon mode interactions. Even though a strong circular dichroism (CD) effect has been predicted in chiral nanostructures with a Fano resonance, there are few experimental studies, and the correlation between the two effects is unclear. In this research, we investigate these two effects in plasmonic heterodimer nanorods in the same spectral range. We find that the heterodimer nanostructure exhibits a Fano-like resonance and Fano-like chiroptical response, both of which are correlated with the coupling between a super-radiant electric dipole and a sub-radiant magnetic dipole mode. Due to the interference nature of the Fano resonance, the Fano-like chiroptical response exhibits distinctively sharp features in a narrow spectral range. This Fano-like chiroptical response can be explained by a modified chiral molecule theory and a simplified coupled electric-magnetic dipole model. This research may provide new insight into the physics picture of plasmonic chirality and paves the way for the development of sensitive plasmonic sensors.

13.
J Phys Chem A ; 124(8): 1659-1665, 2020 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-31994889

RESUMO

Photothermal (PT) microscopy enables the efficient detection of absorbing nano-objects with high sensitivity and stability. The PT signal in the current PT microscopy usually comes from the interaction of the probe laser beam with the heating laser beam-induced thermal lens, and the contribution of the scattering field from the imaged nano-object is usually not taken into account. Here, in this paper, we systematically studied the influence of the scattering field from the imaged nanoparticles on the obtained PT signal by using Ag nanowires (NWs) on a glass substrate surrounded by glycerol as an example. Under the excitation of a heating laser beam at 532 nm wavelength, the rise of local temperature around the Ag NW results in the intensity variation of the interferometric scattering probe light at 730 nm wavelength which includes the scattering light from the Ag NW and the reflection light from the glass-glycerol interface. We found that the PT signal on the NW are positive and negative for the probe beam polarized parallel and perpendicular to the NW axis, respectively. Numerical simulations confirm that the heat-induced intensity variation of the pure scattering light from the NW and the thermal lens-induced intensity increase of the reflection light both contribute to the obtained PT signal. Our work provides the basic guidance for the analysis of PT signal from nano-objects with large scattering cross sections.

14.
Macromol Rapid Commun ; 37(7): 590-6, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26833631

RESUMO

Herein, the use of a 2D soft template system composed of hundred-nanometer-thick water/ethanol mixed layers sandwiched by lamellar bilayer membranes of a self-assembled amphiphilic molecule to produce ultrathin polyprrole (PPy) with a uniform thickness as thin as 3.8 nm and with large dimensions (>2 µm(2)) is presented. The obtained PPy nanosheets exhibit regioregularity with ordered chain alignment where the polymer chains in the nanosheets produced are well aligned with a clear interchain spacing as confirmed by small-angle X-ray scattering measurement. The molecular-level-thick PPy nanosheets exhibit extremely high conductivity up to 1330 S m(-1), thanks to the ordered alignment of polymer chains in the nanosheets, and a high transparency in both the visible region (transmittance >99%) and near-infrared region (transmittance >93%).


Assuntos
Nanoestruturas/química , Polímeros/química , Pirróis/química , Condutividade Elétrica , Microscopia Eletrônica de Varredura , Nanoestruturas/ultraestrutura , Tamanho da Partícula , Espalhamento a Baixo Ângulo , Espectrofotometria , Difração de Raios X
15.
Proc Natl Acad Sci U S A ; 110(12): 4494-9, 2013 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-23487763

RESUMO

Surface plasmons, the quanta of the collective oscillations of free electrons at metal surface, can be easily tuned by changing the surrounding dielectric materials, which is well known for metal nanoparticles and metal surfaces, but less is known for one-dimensional metal nanowires. Here, we find an extremely large tunability of surface plasmons on Ag nanowires with a beat period of the near-field distribution pattern increasing by 90 nm per nanometer of Al2O3 coating, or by 16 µm per refractive index unit change in the surrounding medium. Such high sensitivity is crucial to directly control the optical signal distribution for various routing and demultiplexing functions in plasmonic circuits and may pave the way to the development of on-chip ultrasensitive biosensing.


Assuntos
Elétrons , Nanofios/química , Prata/química , Técnicas Biossensoriais/métodos , Nanofios/ultraestrutura , Propriedades de Superfície
16.
Nano Lett ; 15(1): 560-4, 2015 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-25514318

RESUMO

Plasmonics holds promise for the realization of miniaturized photonic devices and circuits in which light can be confined and controlled at the nanoscale using surface plasmon polaritons (SPPs), surface waves of collective oscillations of electrons at a metal/dielectric interface. However, realizing plasmonic applications fundamentally requires the ability to guide and transfer SPPs in different plasmonic structures. Here the generation and control of periodic collimated SPP-beams are reported in composite structures of silver nanowire on silver film with a dielectric spacer layer between them. It is revealed that the collimated beams on the silver film originate from the interference between film-SPPs generated by two SPP modes on the nanowire. The direction of the collimated beams can be readily tuned by changing the thickness of the dielectric spacer. These findings demonstrate the transfer of nanowire SPPs to film SPPs and offer a new approach to generate nondiffracting SPP-beams, which could facilitate the design and development of complex plasmonic systems for device applications and enable the tailoring of SPP radiation and SPP-matter interactions.

17.
Arch Gerontol Geriatr ; 124: 105441, 2024 09.
Artigo em Inglês | MEDLINE | ID: mdl-38643666

RESUMO

BACKGROUND: The breakdown of the blood-brain barrier (BBB) is intricately linked to the onset and advancement of cognitive impairment and dementia. This investigation explores the correlation between blood-brain barrier permeability, assessed through the cerebrospinal fluid/serum albumin ratio (QAlb), in a clinical cohort and the evolution of cognitive decline. METHODS: This prospective observational cohort study included 295 participants. Cognitive decline progression was characterized by an escalation in the overall deterioration scale and/or clinical dementia rating scores. The investigation delves into the correlation between blood-brain barrier permeability and the advancement of cognitive impairment among patients. RESULTS: The APOE 4 allele and diabetes mellitus among individuals exhibited increased BBB permeability (P < 0.05). Moreover, AD patients exhibited the highest QAlb levels, signifying elevated BBB permeability compared to individuals with MCI and SCD (P < 0.05). After mean 17 months following up, 117 patients (51.31 %) were identified as experiencing cognitive decline progression, and we found that only AD diagnosis, CDR, and QAlb (All P < 0.05) were significant predictors of cognitive decline progression. CONCLUSION: Our study emphasizes the clinical relevance of QAlb in detecting individuals with an elevated risk of cognitive decline. It suggests that heightened BBB permeability could contribute to clinical deterioration and serves as a plausible therapeutic target.


Assuntos
Barreira Hematoencefálica , Disfunção Cognitiva , Progressão da Doença , Humanos , Barreira Hematoencefálica/metabolismo , Masculino , Feminino , Idoso , Estudos Longitudinais , Estudos Prospectivos , Albumina Sérica/metabolismo , Pessoa de Meia-Idade , Fatores de Risco , Permeabilidade , Doença de Alzheimer
18.
Chem Commun (Camb) ; 59(33): 4939-4942, 2023 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-37013669

RESUMO

We develop a new fluorescent biosensor for flap endonuclease 1 (FEN1) assay based on CRISPR/Cas12-enhanced single-molecule counting. This biosensor is simple, selective, and sensitive with a detection limit of 2.325 × 10-5 U and it is applicable for inhibitor screening, kinetic parameter analysis, and quantifying cellular FEN1 with single-cell sensitivity.


Assuntos
Técnicas Biossensoriais , Sistemas CRISPR-Cas , Sistemas CRISPR-Cas/genética , Endonucleases Flap , Cinética , Bioensaio , Corantes
19.
Immun Inflamm Dis ; 11(1): e758, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36705405

RESUMO

INTRODUCTION: To investigate the role of the long-chain noncoding RNA (lncRNA) nuclear enriched abundant transcript 1 (NEAT1) in the process of angiogenesis in human umbilical vein endothelial cells (HUVECs) and illustrate its potential role in burn sepsis (BS) pathogenesis. METHODS: HUVECs were treated with BS patient serum or healthy control serum. NEAT1 shRNA, miR-495-3p mimics, and miR-495-3p inhibitor were transfected into HUVECs. NEAT1 and miR-495-3 levels in serum or HUVECs were detected using quantitative reverse transcription-polymerase chain reaction. Cell counting kit-8 and flow cytometry assays were used to explore the proliferation and apoptosis of HUVECs. The expression of vascular endothelial growth factor (VEGF) in the supernatant was detected using enzyme-linked immunosorbent assay. Tube formation of HUVECs was also analyzed. Western blot analysis was used to analyze signaling pathway proteins. RESULTS: In HUVECs stimulated with BS patient serum, NEAT1 expression was increased, while miR-495-3p expression was decreased. In addition, NEAT1 silencing by specific shRNA inhibited cell proliferation, VEGF production, and tube formation under burn patient serum treatment, which decreased the TGFß1/SMAD signaling pathway activation. Moreover, miR-495-3p minics inhibited angiogenesis and the activation of signaling pathways induced by NEAT1 shRNA. Furthermore, miR-495-3p inhobitor promoted angiogenesis in HUVECs and activated the TGFß1/SMAD signaling pathway. In patients with BS, NEAT1 expression was significantly increased and miR-495-3p expression was decreased compared to healthy controls, and NEAT1 and miR-495-3p expression was associated with the clinical features of patients. CONCLUSIONS: Our results indicate that lncRNA NEAT1 regulates angiogenesis and activates the TGFß1/SMAD signaling pathway during the occurrence of BS.


Assuntos
Queimaduras , MicroRNAs , RNA Longo não Codificante , Sepse , Humanos , Células Endoteliais da Veia Umbilical Humana , MicroRNAs/genética , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , RNA Interferente Pequeno/metabolismo , Sepse/etiologia , Transdução de Sinais , Fator de Crescimento Transformador beta1/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Queimaduras/complicações
20.
Chem Commun (Camb) ; 59(8): 1058-1061, 2023 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-36606583

RESUMO

We demonstrate that target-activated cascade transcription amplification lights up RNA aptamers for label-free detection of metalloproteinase-2 (MMP-2) activity with zero background. This assay exhibits good specificity and high sensitivity with a limit of detection (LOD) of 0.6 fM. Moreover, it can analyze enzyme kinetic parameters, screen inhibitors, and accurately quantify MMP-2 in cancer cells and clinical serums.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Metaloproteinase 2 da Matriz , Aptâmeros de Nucleotídeos/genética , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico
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