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1.
Opt Lett ; 49(13): 3806-3809, 2024 Jul 01.
Article in English | MEDLINE | ID: mdl-38950276

ABSTRACT

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 Jun 28.
Article in English | MEDLINE | ID: mdl-38944964

ABSTRACT

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.

3.
Arch Gerontol Geriatr ; 124: 105441, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38643666

ABSTRACT

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.


Subject(s)
Blood-Brain Barrier , Cognitive Dysfunction , Disease Progression , Humans , Blood-Brain Barrier/metabolism , Male , Female , Aged , Longitudinal Studies , Prospective Studies , Serum Albumin/metabolism , Middle Aged , Risk Factors , Permeability , Alzheimer Disease
4.
Anal Chem ; 95(40): 15133-15139, 2023 10 10.
Article in English | MEDLINE | ID: mdl-37751602

ABSTRACT

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.


Subject(s)
Breast Neoplasms , RNA, Long Noncoding , Humans , Female , Recombinases , Exonucleases , RNA, Long Noncoding/genetics , Breast Neoplasms/diagnosis , Breast Neoplasms/genetics , Nucleotidyltransferases , DNA , Nucleic Acid Amplification Techniques/methods
5.
Anal Chem ; 95(35): 13201-13210, 2023 09 05.
Article in English | MEDLINE | ID: mdl-37603851

ABSTRACT

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.


Subject(s)
Aptamers, Nucleotide , Humans , Streptavidin , Fluorescence Resonance Energy Transfer , RNA Probes , Adenosine , Alpha-Ketoglutarate-Dependent Dioxygenase FTO/genetics
6.
Anal Chem ; 95(31): 11815-11822, 2023 08 08.
Article in English | MEDLINE | ID: mdl-37489894

ABSTRACT

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.


Subject(s)
Biosensing Techniques , Poly(ADP-ribose) Polymerase Inhibitors , Humans , Poly(ADP-ribose) Polymerase Inhibitors/pharmacology , NAD , Static Electricity , Lung
7.
Anal Chem ; 95(20): 8097-8104, 2023 05 23.
Article in English | MEDLINE | ID: mdl-37171156

ABSTRACT

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.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Callosities , RNA, Long Noncoding , Humans , RNA, Long Noncoding/genetics , Zea mays/genetics , Coloring Agents , Limit of Detection
8.
Chem Commun (Camb) ; 59(33): 4939-4942, 2023 Apr 20.
Article in English | MEDLINE | ID: mdl-37013669

ABSTRACT

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.


Subject(s)
Biosensing Techniques , CRISPR-Cas Systems , CRISPR-Cas Systems/genetics , Flap Endonucleases , Kinetics , Biological Assay , Coloring Agents
9.
Chem Commun (Camb) ; 59(8): 1058-1061, 2023 Jan 24.
Article in English | MEDLINE | ID: mdl-36606583

ABSTRACT

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.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Matrix Metalloproteinase 2 , Aptamers, Nucleotide/genetics , Limit of Detection , Nucleic Acid Amplification Techniques
10.
Anal Chem ; 95(5): 3082-3088, 2023 02 07.
Article in English | MEDLINE | ID: mdl-36692970

ABSTRACT

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.


Subject(s)
Neoplasms , RNA, Long Noncoding , Humans , DNA/chemistry , RNA, Long Noncoding/genetics
11.
Immun Inflamm Dis ; 11(1): e758, 2023 01.
Article in English | MEDLINE | ID: mdl-36705405

ABSTRACT

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.


Subject(s)
Burns , MicroRNAs , RNA, Long Noncoding , Sepsis , Humans , Human Umbilical Vein Endothelial Cells , MicroRNAs/genetics , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , RNA, Small Interfering/metabolism , Sepsis/etiology , Signal Transduction , Transforming Growth Factor beta1/metabolism , Vascular Endothelial Growth Factor A/genetics , Burns/complications
12.
Anal Chem ; 94(32): 11425-11432, 2022 08 16.
Article in English | MEDLINE | ID: mdl-35916620

ABSTRACT

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.


Subject(s)
CRISPR-Cas Systems , Neoplasms , Adenosine/metabolism , Alpha-Ketoglutarate-Dependent Dioxygenase FTO/genetics , Alpha-Ketoglutarate-Dependent Dioxygenase FTO/metabolism , Animals , Humans , Mammals/metabolism , Neoplasms/genetics , Obesity/genetics
13.
J Mater Chem B ; 10(17): 3277-3284, 2022 05 04.
Article in English | MEDLINE | ID: mdl-35362489

ABSTRACT

DNA glycosylases are engaged in the base excision repair process and play a vital role in maintaining genomic integrity. It remains a challenge for multiplexed detection of DNA glycosylases in cancer cells. Herein, we demonstrate the construction of a dephosphorylation-mediated chemiluminescent biosensor for multiplexed detection of human alkyladenine DNA glycosylase (hAAG) and uracil DNA glycosylase (UDG) in cancer cells. In this biosensor, the generation of chemiluminescence signals relies on the dephosphorylation of 3-(2'-spiroadamantyl)-4-methoxy-4-(3''-phosphoryloxyphenyl)-1,2-dioxetane (AMPPD) catalyzed by alkaline phosphatase (ALP). We design a bifunctional double-stranded DNA (dsDNA) substrate, a biotin-labelled poly-(T) probe, and two capture probes for the hAAG and UDG assay. This assay involves four steps including (1) the cleavage of the bifunctional dsDNA substrate induced by DNA glycosylases, (2) the recognition of the 3'-OH terminus of the primer by TdT and the subsequent TdT-mediated polymerization reaction, (3) the construction of the AuNPs-dsDNA-ALP nanostructures, and (4) the streptavidin-alkaline phosphatase (SA-ALP)-initiated dephosphorylation of AMPPD for the generation of an enhanced chemiluminescence signal. By taking advantage of the unique features of TdT-mediated polymerization and the intrinsic superiority of the ALP-AMPPD-based chemiluminescence system, this biosensor exhibits good specificity and high sensitivity with a detection limit of 1.53 × 10-6 U mL-1 for hAAG and 1.77 × 10-6 U mL-1 for UDG, and it can even quantify multiple DNA glycosylases at the single-cell level. Moreover, this biosensor can be applied for the measurement of kinetic parameters and the screening of DNA glycosylase inhibitors, holding great potential in DNA damage-related biomedical research and disease diagnostics.


Subject(s)
Biosensing Techniques , Metal Nanoparticles , Neoplasms , Alkaline Phosphatase , DNA/chemistry , Gold , Humans , Uracil-DNA Glycosidase
14.
Small ; 18(43): e2107048, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35229459

ABSTRACT

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.

15.
Anal Chem ; 94(4): 2119-2125, 2022 02 01.
Article in English | MEDLINE | ID: mdl-35050578

ABSTRACT

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.


Subject(s)
Biosensing Techniques , DNA Glycosylases , CRISPR-Cas Systems/genetics , DNA Glycosylases/genetics , DNA Glycosylases/metabolism , DNA Repair , Guanine/analogs & derivatives , Humans
16.
Chem Sci ; 12(31): 10426-10435, 2021 Aug 11.
Article in English | MEDLINE | ID: mdl-34447534

ABSTRACT

MicroRNAs (miRNAs) play key roles in the post-transcriptional regulation of genes, and their aberrant expression may disturb the normal gene regulation network to induce various diseases, and thus accurate detection of miRNAs is essential to early clinical diagnosis. Herein, we develop for the first time a single-quantum dot (QD)-based Förster resonance energy transfer (FRET) nanosensor to accurately detect miRNAs based on copper-free and enzyme-free cycling click chemistry-mediated tricyclic ligase chain reaction (LCR) amplification. We design four DNA probes namely DNA probes 1-4, with DNA probes 1 and 3 being modified with azide (N3) and DNA probes 2 and 4 being modified with dibenzocyclooctyne (DBCO). When target miRNA is present, DNA probes 1 and 2 can proceed via copper-free and enzyme-free click chemistry to generate the probes 1-2 ligation product. Subsequently, DNA probes 3 and 4 can hybridize with the probes 1-2 ligation product to generate the probes 3-4 ligation product. Both the probes 1-2 ligation product and probes 3-4 ligation product can act as the templates to initiate cycling click chemistry-mediated tricyclic LCR amplification whose products can be easily measured by the single-QD-based FRET nanosensor. This assay does not involve any enzymatic reverse transcription, copper catalyst, and ligase enzyme, and it exhibits excellent selectivity, high sensitivity, and the capability of differentiating even single-base mismatches. Moreover, this nanosensor can accurately quantify miRNA-155 even at the single-cell level, and it can distinguish the miRNA-155 expression in tissues of healthy persons and nonsmall cell lung cancer (NSCLC) patients.

17.
Nano Lett ; 21(10): 4193-4201, 2021 05 26.
Article in English | MEDLINE | ID: mdl-33949866

ABSTRACT

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.


Subject(s)
Neoplasms , RNA, Long Noncoding , Neoplasms/diagnosis , Neoplasms/genetics , RNA, Long Noncoding/genetics
18.
Chem Commun (Camb) ; 56(88): 13627-13630, 2020 Nov 14.
Article in English | MEDLINE | ID: mdl-33057479

ABSTRACT

We demonstrate for the first time the integration of a dumbbell probe with dual signal amplification for the simultaneous detection of multiple DNA methyltransferases (MTases). This method is very sensitive with a detection limit of 2.15 × 10-5 U mL-1 for DNA adenine methyltransferase (Dam) and 3.23 × 10-6 U mL-1 for CpG Methyltransferase (M.SssI), and it can be used to screen the enzyme inhibitors and simultaneously measure Dam and M.SssI in complex biological samples.


Subject(s)
DNA Modification Methylases/analysis , DNA Probes/chemistry , A549 Cells , DNA Modification Methylases/metabolism , Humans
19.
Chem Commun (Camb) ; 56(78): 11581-11584, 2020 Oct 01.
Article in English | MEDLINE | ID: mdl-32914789

ABSTRACT

We develop a new fluorescence method for the sensitive detection of protein tyrosine phosphatase 1B (PTP1B) based on dephosphorylation-directed tricyclic DNA amplification cascades. This method exhibits good specificity and high sensitivity with a detection limit of 0.24 pM. Moreover, it can be applied for kinetics analysis, inhibitor screening, and the accurate detection of PTP1B in a variety of cancer cells.


Subject(s)
DNA/metabolism , Nucleic Acid Amplification Techniques/methods , Protein Tyrosine Phosphatase, Non-Receptor Type 1/analysis , Cell Line, Tumor , Chymotrypsin/metabolism , DNA/chemistry , DNA, Catalytic/metabolism , Humans , Limit of Detection , Peptides/chemistry , Peptides/metabolism , Phosphorylation , Substrate Specificity
20.
Ying Yong Sheng Tai Xue Bao ; 31(5): 1487-1495, 2020 May.
Article in Chinese | MEDLINE | ID: mdl-32530225

ABSTRACT

The ENSO events affect climate and fire danger of China. It would be helpful for improving fire danger forecast to understand the impacts of ENSO events on fire weather for various ecological zones in the country. We calculated the fire weather index (FWI) using the daily climatic dataset (V3.0) of international exchange weather stations in China during 1951-2016. The burned areas in forests for each ecological zone in 2001-2016 were derived from MODIS fire products. Temperature, precipitation, FWI and burned areas in fire season were estimated for each ecological zone by ENSO events (weak, medium, strong, and super strong El Niño events and weak, medium, and strong La Niña events). The results showed that there were 19 El Niño events and 14 La Niña events during 1950-2016. The average daily maximum temperature of the spring fire season increased significantly in the northwestern region with the influence of strong or super strong El Niño event, while the temperature reduced significantly in the medium El Niño event for mid-temperate semi-arid grassland. Precipitation in fire season generally increased in El Niño events in southern and southwestern forest regions. It would be reduced in most areas affected by the low and medium intensity La Niña event, but be increased during the strong La Nina event. The fire weather indices of southern forest regions decreased due to the weak El Niño event. The FWI of the northern forest regions increased with the strong or super strong El Niño event, and reduced in the southern and southwestern forest areas. There was a significant spatial difference on the FWI for some ecological zones with the impacts of the El Niño/La Niña events. The burned areas showed a consistent change trend with seasonal severity rating (SSR) during 2001-2016 when the SSR changed significantly for the regions of deciduous broad-leaved forest in humid/semihumid areas of warm temperate zone, broad-leaved forest in the middle north subtropical humid areas, and broad-leaved forest in tropical and subtropical humid areas. The burned areas in the rest regions were not affected by the ENSO events.


Subject(s)
Wildfires , China , El Nino-Southern Oscillation , Seasons , Weather
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