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1.
Stem Cell Res ; 62: 102812, 2022 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-35567849

RESUMO

Dilated cardiomyopathy (DCM) is defined by left ventricular (or biventricular dilation) and systolic dysfunction, which eventually develops into congestive heart failure and arrhythmia. Vinculin is a membrane-associated protein, which functions to transmit contractile force. Defects in vinculin have been reported to be associated with DCM and hypertrophic cardiomyopathy. A human induced pluripotent stem cell (iPSC) line (ZZUNEUi026-A) was generated from a DCM patient carrying heterozygous Vinculin mutant (c.A625 > T; p.Met209 > Leu). The cell line was derived from peripheral blood mononuclear cells by nonintegrative Sendai virus. ZZUNEUi026-A showed pluripotency markers and normal karyotype, and it could differentiate into three germ layers in vitro.

2.
Cancer Cell Int ; 22(1): 191, 2022 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-35578338

RESUMO

BACKGROUND: Deacetylation of histones by histone deacetylase 3 (HDAC3) acts importantly in modulating apoptosis, DNA damage and cellular progression. Herein, we aimed to unravel the functional role of HDAC3 in a lethal disease, esophageal squamous cell carcinoma (ESCC). METHODS: The expression of HDAC3 in clinically collected ESCC tissues was determined by RT-qPCR and immunohistochemistry. As revealed from bioinformatics analysis, the putative relations between HDAC3 and microRNA-494 (miR-494) and between miR-494 and transforming growth factor beta (TGFß)-inducing factor 1 (TGIF1) were further verified by chromatin immunoprecipitation and dual-luciferase reporter gene assay. Functional roles of shRNA-mediated depletion of HDAC3, miR-494 mimic and overexpressed TGIF1 were explored by gain- and loss-of-function assays with regard to ESCC cell biological behaviors. A nude mouse model of ESCC was developed for in vivo validation. RESULTS: HDAC3 was highly expressed in ESCC tissues, suggestive of poor prognosis while TGIF1 was upregulated and miR-494 was downregulated. Mechanistic investigation revealed that HDAC3 inhibited miR-494 expression and TGIF1 was a direct target of miR-494. Furthermore, silencing HDAC3 or overexpressing miR-494 was demonstrated to suppress aggressive phenotypes of ESCC cells both in vitro through the activated TGFß signaling pathway and in vivo, while TGIF1 overexpression induced opposite results. CONCLUSION: Collectively, our findings provided demonstration regarding the oncogenic property of HDAC3 in ESCC via the miR-494/TGIF1/TGFß axis.

3.
J Clin Psychol ; 2022 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-35545873

RESUMO

OBJECTIVES: The protective role of self-compassion in emerging adult depression has garnered empirical support. It makes more sense to understand the psychological processes underlying this relationship. Based on the stress appraisal patterns, the present study examined the mediating roles of emotion regulation (ER) and resilience in the link between self-compassion and depression among college students with left-behind experience (LBE). METHODS: A total of 391 LBE college students (Mage = 18.43 years; SD = 0.79 years) in Chongqing reported their demographic information and self-compassion (the Self-Compassion Scale) level at baseline (T1) and reported their levels of ER (the Emotion Regulation Scale), resilience (the Connor-Davidson Resilience Scale), and depression (the Depression-Anxiety-Stress Scale) 3 months later (T2). RESULTS: The results revealed that (a) both ER (cognitive reappraisal and expressive suppression) and resilience separately mediated the association between self-compassion and depression; (b) cognitive reappraisal and resilience sequentially mediated this association. CONCLUSIONS: These findings reveal the underlying mechanisms of the associations between self-compassion and depression among LBE college students and have implications for interventions aimed at mitigating their depression.

4.
ACS Omega ; 7(12): 10365-10371, 2022 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-35382338

RESUMO

Two-dimensional (2D) and quasi-2D Ruddlesden-Popper (RP) phase organolead halide perovskites are promising materials for both photovoltaic and optoelectronic devices. Although they are known to be more stable when exposed to moisture than their 3D counterpart, chemical degradation of these materials under moisture, which not only leads to a significant drop in device performance but also leads to lead leakage, yet remains one of the most serious hurdles for their practical applications. To gain insight into the degradation mechanism of 2D/quasi-2D perovskites under moisture conditions, the degradation pathway of 2D/quasi-2D (PEA)2(MA) n-1PbnI3n+1 (PEA = C6H5C2H4NH3 +, MA = CH3NH3 +, and n is the number of perovskite layers between adjacent organic spacer layers) perovskite single crystals (SCs) and thin film are explored. We observe the degradation process by mapping the photoluminescence of the 2D perovskites and demonstrate that the larger-n phases all directly degrade into the relative stable n = 1 phase and MAI and PbI2, which is a mechanism different from that in previous reports and further confirmed in the 2D perovskite thin film. This degradation process is also found to be independent of the boundary and morphology of the SCs. This discovery provides a new perspective for understanding the chemical degradation of the 2D perovskite materials and may inspire new solutions for improving their moisture stability.

5.
Polymers (Basel) ; 14(8)2022 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-35458346

RESUMO

Organogel adsorbents are widely used for the adsorption of hard-to-degrade organic pollutants in wastewater due to their natural affinity to the organic phase in water. In this study, phenolic xerogels (PF) synthesised in the ethylene glycol inorganic acid system are used as a backbone and superhydrophobic phenolic xerogels (ASO-PF) are obtained by grafting aminosilanes onto the PF backbone via the Mannich reaction. The modified ASO-PF not only retains the pore structure of the original PF (up to 90% porosity), but also has excellent superhydrophobic properties (water contact angle up to 153°). Owing to the unique pore structure, ASO-PF has excellent compression properties, cycling 50% compression deformation more than 10 times without being damaged, with a maximum compression deformation of up to 80%. A squeeze-suction-squeeze approach is proposed for selective adsorption of organic pollutants in homogeneous solutions based on the recyclable compression properties of ASO-PF. The ASO-PF is put under negative pressure by squeezing, and when the pressure is released, the adsorbed liquid enters the ASO-PF, where the organic pollutants are retained by the adsorption sites in the skeleton, and then the remaining water is discharged by squeezing. This breathing ASO-PF holds great promise for organic pollutant adsorption and recovery applications.

6.
Ann Transl Med ; 10(6): 279, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35433956

RESUMO

Background: Emerging evidence suggests that secreted phosphoprotein 1 (SPP1) is involved in tumor cell progression in multiple cancer types. However, the role of SPP1 in different cancers is still not clear. Methods: We used data from The Cancer Genome Atlas (TCGA) to analyze the multiomic roles of SPP1, including RNA expression, DNA methylation, protein phosphorylation, immune infiltration, and overall survival (OS) in 33 tumor types. Results: SPP1 is highly expressed in most cancer types, and its methylation variability and mRNA expression level are both correlated with prognosis in multiple cancer types. A higher S234 phosphorylation level was observed in 4 types of tumors, including colon adenocarcinoma (COAD) and lung adenocarcinoma (LUAD). SPP1 expression level was positively associated with the infiltration level of dendritic cells, neutrophils, and macrophages in multiple cancer types. It was also significantly positively correlated with hepatitis A virus cellular receptor 2 (HAVCR2), which was observed in most tumor types, including brain lower grade glioma (LGG) and ovarian serous cystadenocarcinoma (OV). Moreover, myeloid cell differentiation and leukocyte migration were observed in the enrichment analysis, suggesting that SPP1 might induce immune escape. Conclusions: Pan-cancer analysis using a multiomic approach offered a comprehensive overview of SPP1. This protein plays an important role in most of the analyzed tumor types and could be a valuable prognostic marker across different types of cancer.

7.
Anal Chem ; 94(15): 6004-6010, 2022 04 19.
Artigo em Inglês | MEDLINE | ID: mdl-35384669

RESUMO

Alkaline phosphatase (ALP) is a valuable biomarker and effective therapeutic target for the diagnosis and treatment of diverse human diseases, including bone disorder, cardiovascular disease, and cancers. The reported ALP assays often suffer from laborious procedures, costly reagents, inadequate sensitivity, and large sample consumption. Herein, we report a new single-molecule fluorescent biosensor for the simple and ultrasensitive detection of ALP. In this assay, the ALP-catalyzed dephosphorylation of detection probe can protect the detection probe against lambda exonuclease-mediated digestion, and the remaining detection probes can trigger ceaseless hybridization between two Cy5-labeled hairpin probes through toehold-mediated DNA strand displacement, generating a long fluorescent DNA chain, which can be subsequently separated from unhybridized hairpin probes and disassembled into dispersed Cy5 moieties upon NaOH treatment. The free Cy5 moieties indicate the presence of ALP and can be directly quantified via single-molecule counting. This biosensor enables efficient amplification and transduction of the target ALP signal through enzyme-free assembly and disassembly processes, significantly simplifying the experimental procedure and improving the assay accuracy. The proposed biosensor allows specific and ultrasensitive detection of ALP activity with a detection limit down to 2.61 × 10-6 U mL-1 and is suitable for ALP inhibition assay and kinetic analysis. Moreover, this biosensor can be applied for endogenous ALP detection in human cells and clinical human serum, holding the potential in the ALP biological function study and clinical diagnosis.


Assuntos
Fosfatase Alcalina , Técnicas Biossensoriais , Fosfatase Alcalina/metabolismo , Catálise , Corantes , DNA , Humanos , Cinética , Limite de Detecção
8.
Anal Chem ; 94(15): 5980-5986, 2022 04 19.
Artigo em Inglês | MEDLINE | ID: mdl-35394287

RESUMO

DNA methylation is an essential genomic epigenetic behavior in both eukaryotes and prokaryotes. Deregulation of DNA methyltransferase (Dam MTase) can change the DNA methylation level and cause various diseases. Herein, we develop an apurinic/apyrimidinic endonuclease 1 (APE1)-mediated cascade signal amplification platform for homogeneously sensitive and rapid measurement of Dam MTase in Escherichia coli cells. This assay involves a partial double-stranded DNA (dsDNA) substrate and two hairpin signal probes (HP1 and HP2) that are modified with Cy5 and BHQ2 at two ends, respectively. When Dam MTase is present, it methylates the dsDNA substrate, and subsequently, endonuclease DpnI cleaves the methylated substrate, yielding trigger probe 1. Hybridization of trigger probe 1 with HP1 forms a partial dsDNA containing an apurinic/apyrimidinic (AP) site, which is cleaved by APE1 to induce the cyclic cleavage of HP1 and the production of abundant trigger probe 2. Subsequent hybridization of trigger probe 2 with HP2 forms a partial dsDNA with an AP site, inducing the cyclic cleavage of HP2 by APE1. Consequently, cyclic cleavage of HP1 and HP2 induces the generation of abundant Cy5 molecules, which are easily measured by single-molecule imaging. This assay can be performed homogeneously and rapidly within 2 h, which is the shortest among the reported amplification-based assays. Moreover, it exhibits good selectivity and high sensitivity, and it can discriminate Dam MTase from other enzymes and screen inhibitors. Importantly, it can accurately measure the Dam MTase activity in serum and E. coli cells, with promising applications in clinical diagnosis and drug discovery.


Assuntos
Técnicas Biossensoriais , DNA Metiltransferases Sítio Específica (Adenina-Específica) , Técnicas Biossensoriais/métodos , Proteínas Cromossômicas não Histona , DNA , Metilação de DNA , Metilases de Modificação do DNA , Endonucleases , Escherichia coli/metabolismo , DNA Metiltransferases Sítio Específica (Adenina-Específica)/metabolismo
9.
J Mater Chem B ; 10(17): 3277-3284, 2022 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-35362489

RESUMO

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.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Neoplasias , Fosfatase Alcalina , DNA/química , Ouro , Humanos , Uracila-DNA Glicosidase
10.
J Mater Chem B ; 10(17): 3260-3267, 2022 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-35377384

RESUMO

Protein kinase can catalyze the phosphorylation of peptides/proteins, and it is closely associated with various human diseases such as cancer, immune deficiencies, and Alzheimer's disease. Sensitive monitoring of protein kinase activity is significant for biochemical research and drug discovery. Herein, we develop a phos-tag-based fluorescent biosensor for sensitive detection of cAMP-dependent protein kinase (PKA) activity in cancer cells. We design a peptide-DNA conjugate and a signal probe for PKA activity assay, and employ a biotinylated phos-tag (a selective phosphate-binding agent) to recognize and capture the phosphorylated peptide-DNA substrate. The peptide-DNA conjugates as the catalytic substrates can translate the peptide signal to a nucleotide signal for the initiation of the RNase HII-driven cycling signal amplification. The magnetic nanobeads (MBs) as the carriers can separate and enrich the phosphorylated peptide in complex matrices. Meanwhile, the combination of MBs with the phos-tag-mediated phosphate group recognition can effectively eliminate the interference from the complex matrix, and the introduction of single-molecule detection endows this assay with high sensitivity. This biosensor can achieve a detection limit of 1.98 × 10-8 U µL-1 and a wide dynamic range from 1 × 10-7 to 1 × 10-2 U µL-1. Moreover, this biosensor can be applied for the screening of PKA inhibitor and the measurement of cellular PKA activity, holding great potential in biomedical research and clinical diagnosis.


Assuntos
Técnicas Biossensoriais , Neoplasias , Humanos , Peptídeos/química , Fosfatos , Proteínas Quinases/metabolismo , Piridinas
11.
Chem Commun (Camb) ; 58(36): 5538-5541, 2022 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-35436780

RESUMO

We develop a dual-functional dumbbell probe-based fluorescent biosensor for cascade amplification detection of miRNAs in lung cancer cells and tissues by integrating a primer exchange reaction (PER) with the CRISPR-Cas12a system. This biosensor can absolutely quantify the miR-486-5p expression in different lung cancer cells and distinguish non-small cell lung cancer (NSCLC) patients from healthy individuals, holding great potential in biomedical research and clinical diagnosis.


Assuntos
Técnicas Biossensoriais , Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , MicroRNAs , Humanos , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/genética , MicroRNAs/genética
12.
Exp Ther Med ; 23(5): 344, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35401796

RESUMO

Arctium lappa L., also known as burdock, is a popular medicinal plant in traditional Chinese medicine due to its potential therapeutic properties. Saccharides from Arctium lappa L. root (ALR-S) have been extensively studied for their anti-inflammatory and anti-diabetes effects. Platelets play a pivotal role in thrombosis. The present study describes the effects of ALR-S on platelet activation and thrombosis using a laser injury thrombosis in vivo model. The study also measured the effects of ALR-S on platelet activation by analysing aggregation, ATP release, platelet spreading, adhesion and clot retraction in vitro. Specifically, the effects were ALR-S concentration-dependent inhibition of platelet aggregation and ATP release. Activated platelets pretreated with ALR-S showed diminished CD62P expression levels and fibrinogen binding, as measured by flow cytometry. ALR-S inhibited platelet spreading on fibrinogen and adhesion on collagen under shear. ALR-S attenuated platelet activation by decreasing oxidative stress and thrombus formation. These results demonstrated the antiplatelet effects of ALR-S, suggesting the antithrombotic and cardiovascular protective activities of ALR-S as a functional food.

13.
Talanta ; 243: 123342, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35255432

RESUMO

The structure-specific endonuclease FEN1 participates in various genome maintenance pathways in eukaryotes and is associated with different human diseases. Herein, we demonstrate label-free and homogeneous detection of FEN1 based on ligation-promoted hyperbranched rolling circle amplification (HRCA). This assay can be performed isothermally with the involvement of primers 1 and 2 and a circular DNA substrate with a 5'-flap. When FEN1 is present, it cleaves 5'-flap of circular DNA substrate to obtain a circular padlock probe with the assistance of Taq DNA ligase. The circular padlock probe can serve as a template to initiate HRCA in the presence of primers 1 and 2 and Vent (exo-) DNA polymerase. The obtained dsDNA fragments can produce an enhanced fluorescence signal with SYBR Green I as indicator. This method displays good specificity and high sensitivity, and it can be employed to screen FEN1 inhibitors and quantitatively detect FEN1 activity in human cancer cells, with potential applications in early diagnosis and drug discovery.


Assuntos
Endonucleases Flap , Técnicas de Amplificação de Ácido Nucleico , DNA/genética , DNA Ligases , Humanos
14.
Talanta ; 243: 123340, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35272158

RESUMO

Guanine is the most susceptible to oxidation among all the DNA bases, and 8-oxo-7,8-dihydroguanine (OG) is one of main oxidation products that can occur in any part of chromosomal DNA. OG in the telomere sequence is associated with telomere shortening, cell aging, and dysfunction, and it may induce cancers. The accurate detection of OG in telomeres is important to early clinical diagnosis and molecular research. Herein, we develop a simple and rapid method to sensitively measure 8-oxo-7,8-dihydroguanine (OG) in telomeres of cancer cells by using Bsu polymerase-mediated fluorescence coding. This method is very simple without the requirement for any nucleic acid amplification or specific restriction enzyme recognition reaction, and Bsu polymerase can selectively incorporate Cy5-dATP into the opposite site of OG, endowing this method with good specificity. Moreover, the introduction of single-molecule detection significantly improves the sensitivity. This method can detect OG within 70 min with a limit of detection (LOD) of 2.45 × 10-18 M, and it can detect OG in genomic DNA extracted from H2O2-treated HeLa cells with a LOD of 0.0094 ng, holding great potential in disease-specific gene damage research and early clinic diagnosis.


Assuntos
Peróxido de Hidrogênio , Neoplasias , Dano ao DNA , Fluorescência , Guanina/análogos & derivados , Células HeLa , Humanos , Neoplasias/diagnóstico , Neoplasias/genética , Oxirredução , Telômero/genética
15.
Trauma Violence Abuse ; : 15248380211073845, 2022 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-35258353

RESUMO

Objectives: Intimate partner violence (IPV) against pregnant or human immunodeficiency virus (HIV)-positive women have been previously studied. However, data on the impact of IPV on HIV-positive pregnant women have not been systematically synthesized. We performed a meta-analysis to explore this issue and provide evidence regarding IPV prevention and HIV infection control. Method: The PubMed, Web of Science, Cochrane Library, and Embase databases were systematically searched. Studies that quantitatively assessed the association between IPV and its adverse impact on HIV-positive women during pregnancy and post-partum were eligible for inclusion. Pooled odds ratios (ORs) were calculated. Findings: Eight studies were identified to meet our eligibility criteria. The adverse impacts of IPV against HIV-positive pregnant women mainly included nonadherence to maternal antiretroviral treatment during pregnancy, nondisclosure of HIV-positive status to male partners, nonadherence to infant antiretroviral prophylaxis, and antenatal depression. IPV caused a 180% and 145% increase in the odds of antenatal depression and nonadherence to infant antiretroviral prophylaxis, respectively, among HIV-positive women, compared to the odds of their IPV-free counterparts [OR = 2.80, 95% confidence interval (CI): 1.66-4.74; OR = 2.45, 95% CI: 1.40-4.27]. Conclusion: Limited evidence has suggested that IPV against HIV-positive pregnant women caused maternal depression during pregnancy and led to the possible failure of HIV prophylaxis adherence in infants. Interventions to address IPV may ultimately reduce the risk of depression-related adverse birth outcomes and vertical transmission in infants exposed to maternal HIV. Prevention and control against IPV should be developed for HIV-positive pregnant women.

16.
Ren Fail ; 44(1): 146-154, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35164637

RESUMO

OBJECTIVES: To analyze conventional ultrasound (CUS) and contrast-enhanced ultrasound (CEUS) features in patients with secondary hyperparathyroidism (SHPT) and to evaluate the clinical-ultrasonographic feature based model for predicting the severity of SHPT. METHODS: From February 2016 to March 2021, a total of 59 patients (age 51.3 ± 11.7 years, seCr 797.8 ± 431.7 µmol/L, iPTH 1535.1 ± 1063.9 ng/L) with SHPT (including 181 parathyroid glands (PTGs)) without the history of intact parathyroid hormone (iPTH)-reducing drugs using were enrolled. The patients were divided into the mild SHPT group (mSHPT, iPTH <800 ng/L) and the severe SHPT group (sSHPT, iPTH ≥ 800 ng/L) according to the serum iPTH level. The clinical test data of patients were collected and CUS and CEUS examinations were performed for every patient. Multivariable logistic regression model according to clinical-ultrasonographic features was adopted to establish a nomogram. We performed K-fold cross-validation on this nomogram model and nomogram performance was determined by its discrimination, calibration, and clinical usefulness. RESULTS: There were 19 patients in the mSHPT group and 40 patients in the sSHPT group. Multivariable logistic regression indicated serum calcium, serum phosphorus and total volume of PTGs were independent predictors related with serum iPTH level. Even though CEUS score of wash-in and wash-out were showed related to severity of SHPT in univariate logistic regression analysis, they were not predictors of SHPT severity (p = 0.539, 0.474 respectively). The nomogram developed by clinical and ultrasonographic features showed good calibration and discrimination. The accuracy and the area under the curve (AUC), positive predictive value (PPV), negative predictive value (NPV) and accuracy of this model were 0.888, 92.5%, 63.2% and 83.1%, respectively. When applied to internal validation, the score revealed good discrimination with stratified fivefold cross-validation in the cohort (mean AUC = 0.833). CONCLUSIONS: The clinical-ultrasonographic features model has good performance for predicting the severity of SHPT.


Assuntos
Hiperparatireoidismo Secundário/diagnóstico por imagem , Falência Renal Crônica/complicações , Glândulas Paratireoides/diagnóstico por imagem , Diálise Renal/efeitos adversos , Ultrassonografia Doppler em Cores/métodos , Adulto , Idoso , Cálcio/sangue , Feminino , Humanos , Hiperparatireoidismo Secundário/sangue , Hiperparatireoidismo Secundário/etiologia , Hiperparatireoidismo Secundário/cirurgia , Falência Renal Crônica/metabolismo , Falência Renal Crônica/terapia , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Nomogramas , Glândulas Paratireoides/irrigação sanguínea , Glândulas Paratireoides/patologia , Hormônio Paratireóideo/sangue , Paratireoidectomia , Fósforo/sangue , Estudos Retrospectivos , Resultado do Tratamento
17.
Comput Math Methods Med ; 2022: 8354932, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35047058

RESUMO

METHODS: The gene expression data were extracted from the "Therapeutically Applicable Research to Generate Effective Treatments" (TARGET) database. The differentially expressed genes (DEGs) were identified, and the relationships between DEGs and m6A genes were explored. Then, the correlations among the m6A genes in neuroblastoma were investigated. Finally, the prognostic role of the m6A genes was studied, and the risk model was constructed. RESULTS: 81 NB patients were extracted from the TARGET database. After comparing the gene expression between unfavorable and favorable outcome groups, 73 DEGs were identified, including 54 downregulated genes and 19 upregulated genes. In NB patients, we found that IGF2BP3, METTL14, and METTL16 are prognostic factors for disease-free survival (DFS) while IGF2BP3, METTL14, and METTL16 are prognostic factors for overall survival (OS). Besides, after the risk model construction, the OS between the two risk groups was drawn (log-rank p = 1.64e - 08, HR = 3.438, 95% CI 2.24-5.278). The 1-, 3-, and 5-year time-dependent receiving operating characteristic (ROC) curves were also illustrated, and the areas under the receiver operating characteristic curves (AUCs) attained 0.75, 0.798, and 0.768, respectively. CONCLUSIONS: IGF2BP3, METTL14, and METTL16 were identified as the significant factors for DFS and OS in NB patients.


Assuntos
Metiltransferases/genética , Neuroblastoma/genética , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Criança , Biologia Computacional , Bases de Dados Genéticas , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Humanos , Estimativa de Kaplan-Meier , Metilação , Metiltransferases/metabolismo , Neuroblastoma/metabolismo , Prognóstico , Mapas de Interação de Proteínas/genética , Processamento Pós-Transcricional do RNA/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Fatores de Risco
18.
Anal Chem ; 94(3): 1882-1889, 2022 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-35000391

RESUMO

MicroRNAs (miRNAs) play multiple crucial roles in post-transcriptional regulating gene expression, and the abnormal expression may induce various human diseases. Herein, we demonstrate the construction of a structure-switchable toehold dumbbell probe for sensitive and label-free measurement of microRNA in cancer cells and tissues on the basis of integrating exponential-rolling circle amplification (EXP-RCA) with linear-rolling circle amplification (LRCA). We designed a structure-switchable toehold dumbbell probe with annular and symmetric structure whose either side can hybridize with target miRNA to initiate EXP-RCA, greatly improving the detection sensitivity. Moreover, the dumbbell probe is designed with an appropriate standard free energy (G), and it cannot be activated by mismatched miRNAs, endowing this assay with good specificity. When target miRNA is present, it interacts with the dumbbell probe to activate EXP-RCA via toehold-mediated stand displacement, generating abundant triggers. The resulting triggers and target miRNA can function as primers to initiate LRCA, producing abundant long tandem repeats that can generate a distinct fluorescence signal using SYBR Gold as the indicator. This assay can be carried out homogeneously and isothermally without the requirement for either sophisticated modification/separation steps or any extra primers. It displays ultrahigh sensitivity with a limit of detection of 8.45 × 10-17 M and excellent specificity, and it can differentiate let-7a from its homologous analogues. Moreover, this method can accurately quantify let-7a expression at a single-cell level and can even distinguish the let-7a expression between non-small cell lung cancer patient tissues and healthy person tissues.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , MicroRNAs , Primers do DNA , Humanos , Limite de Detecção , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/genética , MicroRNAs/genética , Técnicas de Amplificação de Ácido Nucleico/métodos
19.
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
20.
Chem Commun (Camb) ; 58(10): 1565-1568, 2022 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-35014995

RESUMO

We develop for the first time a label-free fluorescent method for sensitive detection of fat mass and obesity-associated protein (FTO) activity using MazF-mediated primer generation rolling circle amplification. This method is very simple with ultrahigh sensitivity and good specificity, and it can detect FTO activity at the single-cell level. Moreover, this method can be applied for the measurement of kinetic parameters and the screening of FTO inhibitors.


Assuntos
Dioxigenase FTO Dependente de alfa-Cetoglutarato/análise , DNA de Cadeia Simples/metabolismo , Técnicas de Amplificação de Ácido Nucleico/métodos , Dioxigenase FTO Dependente de alfa-Cetoglutarato/metabolismo , Linhagem Celular Tumoral , DNA de Cadeia Simples/química , Proteínas de Ligação a DNA/metabolismo , Endorribonucleases/metabolismo , Proteínas de Escherichia coli/metabolismo , Humanos , Limite de Detecção , Eletroforese em Gel de Poliacrilamida Nativa , Análise de Célula Única , Espectrometria de Fluorescência
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