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1.
Appl Spectrosc ; : 37028241238246, 2024 Mar 26.
Article in English | MEDLINE | ID: mdl-38529537

ABSTRACT

The precise regulation of nanoenzyme activity is of great significance for application to biosensing analysis. Herein, the peroxidase-like activity of carbon dots was effectively modulated by doping phosphorus, which was successfully employed for sensitive, selective detection of acid phosphatase (ACP). Phosphorus-doped carbon dots (P-CDs) with excellent peroxidase-like activity were synthesized by a one-pot hydrothermal method, and the catalytic activity could be easily modulated by controlling the additional amount of precursor phytic acid. P-CDs could effectively catalyze the oxidation of colorless 3,3',5,5'-tetramethylbenzidine (TMB) to blue TMB oxidation products in the presence of hydrogen peroxide. While ACP was able to catalyze the hydrolysis of L-ascorbyl-2-phosphate trisodium salt (AAP) to produce ascorbic acid (AA), which inhibited the peroxidase-like activity of P-CDs, by combining P-CDs nanoenzymes and ACP-catalyzed hydrolysis the colorimetric method was established for ACP detection. The absorbance variation showed a good linear relationship with ACP concentration in the range of 0.4-4.0 mU/mL with a limit of detection at 0.12 mU/mL. In addition, the method was successfully applied to detect ACP in human serum samples with recoveries in the range of 98.7-101.6%. The work provides an effective strategy for regulating nanoenzymes activity and a low-cost detection technique for ACP.

2.
Spectrochim Acta A Mol Biomol Spectrosc ; 314: 124192, 2024 Jun 05.
Article in English | MEDLINE | ID: mdl-38552541

ABSTRACT

Catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) can achieve the high sensitivity and rapid reaction rate in detecting miRNA. However, the amplification efficiency by these methods are limited. Herein, an enzyme-free and label-free hyperbranched DNA network structure (HDNS) was designed, in which localized catalytic hairpin assembly (LCHA) and hybridization chain reaction occurred in the horizontal axis and longitudinal axis, respectively, exhibiting intensive signal dual-amplification. miRNA-122 was selected as the target on behalf of miRNA to design the HDNS sensor. The fluorescence signal change of HDNS showed good linearity for detecting miRNA-122 in the concentration range from 0.1 nM to 60 nM with a limit of detection (LOD) at 37 pM which was lower than those of the sensors based on separate CHA or HCR. Afterwards, the HDNS sensor was applied to detect miRNA-122 in serum samples with the recovery rate in the range of 97.2 %-107 %. The sensor could distinguish different kinds of miRNAs, even the family members with high sequence homology, exhibiting excellent selectivity. This method provided a novel design strategy for improving the sensitivity and selectivity of DNA sensor for miRNA detection.


Subject(s)
Biosensing Techniques , MicroRNAs , MicroRNAs/genetics , Biosensing Techniques/methods , DNA/chemistry , Nucleic Acid Hybridization/methods , Limit of Detection
3.
Exp Ther Med ; 26(2): 384, 2023 Aug.
Article in English | MEDLINE | ID: mdl-37456164

ABSTRACT

The aim of the present study was to explore the association between anemia and the risks and outcomes of diabetic foot (DF) in patients with type 2 diabetes mellitus (T2DM). A total of 145 patients with T2DM were recruited between January and December 2021 and divided into the DF and non-DF groups according to whether they were diagnosed with DF. Individual patient data were extracted and blood samples were evaluated in a biochemical center for routine biochemical and blood-related indicators. The patients' survival rates were followed up until December 2022. An independent-samples t-test and χ2 test were used to compare the differences between the two groups. The association between the various clinical indicators for the DF and non-DF groups was evaluated using single-factor binary logistic regression analysis. Multi-factor binary logistic regression analysis was used to analyze the association between hemoglobin (Hb) and the risk for DF. A Kaplan-Meier survival curve was used to analyze the impact of anemia and DF on the 1-year survival rate. The diabetes duration, number of patients who smoked and consumed alcohol, and serum creatinine and C-reactive protein levels in the DF group were significantly higher than those in the non-DF group (P<0.05). By contrast, the estimated glomerular filtration rate (eGFR) and Hb, albumin (Alb) and total cholesterol levels, were lower in the DF group when compared with those in the non-DF group (P<0.05). All of the study participants were divided into two groups, according to their baseline eGFR [eGFR ≥90 or <90 ml/(min x 1.73 m²)]. It was found that, independently of renal function, lower Hb and Alb levels were associated with a higher incidence of DF. The 1-year survival rate for DF with anemia was significantly lower when compared with that in patients with DF without anemia (P<0.05). In conclusion, the Hb level in patients with T2DM is a protective factor against DF and anemia is an independent risk factor for DF. The present study suggested that anemia is associated with a decrease in the survival rate of patients with DF. This finding provided a theoretical basis for the clinical correction of anemia and improvement of DF prognosis (clinical trial no. 20220003).

4.
Curr Issues Mol Biol ; 45(7): 5598-5612, 2023 Jun 30.
Article in English | MEDLINE | ID: mdl-37504269

ABSTRACT

The pathogenesis of head and neck squamous cell carcinoma (HNSCC) is associated with human papillomavirus (HPV) infection. However, the molecular mechanisms underlying the interactions between HNSCC and HPV remain unclear. Bioinformatics was used to analyze the gene expression dataset of HPV-associated HNSCC based on the Cancer Genome Atlas (TCGA) database. Differentially expressed genes (DEGs) in HPV-positive and HPV-negative HNSCC were screened. Gene function enrichment, protein-protein interactions (PPI), survival analysis, and immune cell infiltration of DEGs were performed. Furthermore, the clinical data of HNSCC tissue samples were analyzed using immunohistochemistry. In total, 194 DEGs were identified. A PPI network was constructed and 10 hub genes (EREG, PLCG1, ERBB4, HBEGF, ZFP42, CBX6, NFKBIA, SOCS1, ATP2B2, and CEND1) were identified. Survival analysis indicated that low expression of SOCS1 was associated with worse overall survival. Immunohistochemistry demonstrated that SOCS1 expression was higher in HPV-negative HNSCC than in HPV-positive HNSCC, and there was a positive correlation between SOCS1 expression and patient survival. This study provides new information on biological targets that may be relevant to the molecular mechanisms underpinning the occurrence and development of HNSCC. SOCS1 may play an important role in the interaction between HPV and HNSCC and serve as a potential biomarker for future therapeutic targets.

5.
Arch. endocrinol. metab. (Online) ; 67(3): 306-313, June 2023. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1429759

ABSTRACT

ABSTRACT Objective: Intrathyroidal injection using an insulin pen filled with a mixture of lidocaine and triamcinolone acetonide is a therapy for subacute thyroiditis (SAT) reported by us previously. We aimed to evaluate the clinical efficacy of ultrasound-guided intrathyroidal injection in the treatment of SAT. Subjects and methods: A total of 93 patients with SAT completed the study. All patients were evaluated via a history and clinical examination followed by thyroid function tests and ultrasonography of the thyroid. After ultrasound-guided intrathyroidal injection, the patients were followed up with respect to the injection frequency, treatment duration, and patient satisfaction. The visual numerical rating scale was used as a pain questionnaire for a given interval. Results: Thyroid pain instantly decreased to scores below 3.0 following the first injection. Sixty-three patients (67.74%) avoided relapse of thyroid pain within 3 injections, which occurred within only 3 days after the first injection. The pain in 27 patients (29.03%) disappeared completely after 4-6 injections. Only 3 patients (3.23%) were found to need more than 6 injections, with 10 cited as the maximum number of injections, the injection took only 17 days altogether. The mean treatment cycle of the intrathyroidal injection was 3.98 days. There were no other associated complications with the novel therapy except infrequent small subcutaneous hematomas, which could be prevented with skilled practice. The average patient satisfaction score was as high as 9.0. Conclusion: Intrathyroidal injection of lidocaine and triamcinolone acetonide using an insulin pen was found to be an advantageous and satisfactory treatment for SAT.

6.
Talanta ; 259: 124533, 2023 Jul 01.
Article in English | MEDLINE | ID: mdl-37058942

ABSTRACT

Deoxyribonuclease I (DNase I) is a typical nuclease that plays key roles in many physiological processes and the development of a novel biosensing strategy for DNase I detection is of fundamental significance. In this study, a fluorescence biosensing nanoplatform based on a two-dimensional (2D) titanium carbide (Ti3C2) nanosheet for sensitive and specific detection of DNase I was reported. Fluorophore-labeled single-stranded DNA (ssDNA) can be spontaneously and selectively adsorbed on Ti3C2 nanosheet through the hydrogen bond and metal chelate interaction between phosphate groups of ssDNA and titanium of Ti3C2 nanosheet, resulting in effective quenching of the fluorescence emitted by fluorophore. Notably, it was found the enzyme activity of DNase I will be terminated by the Ti3C2 nanosheet. Therefore, the fluorophore-labeled ssDNA was firstly digested by DNase I and the "post-mixing" strategy of Ti3C2 nanosheet was chosen to evaluate the enzyme activity of DNase I, which provided the possibility of improving the accuracy of the biosensing method. Experimental results demonstrated that this method can be utilized for quantitative analysis of DNase I activity and exhibited a low detection limit of 0.16 U/ml. Additionally, the evaluation of DNase I activity in human serum samples and the screening of inhibitors with this developed biosensing strategy were successfully realized, implying that it has high potential as a promising nanoplatform for nuclease analysis in bioanalytical and biomedical fields.


Subject(s)
Biosensing Techniques , Titanium , Humans , Titanium/chemistry , Biosensing Techniques/methods , DNA, Single-Stranded , Fluorescent Dyes/chemistry , Deoxyribonuclease I
7.
Mikrochim Acta ; 190(4): 149, 2023 03 23.
Article in English | MEDLINE | ID: mdl-36952059

ABSTRACT

An electrochemical strategy based on bimetallic nanozyme in collaboration with toehold-mediated DNA replacement effect is proposed for the sensitive determination of miRNA-21. The AuPt nanoparticles (AuPt NPs) are prepared as a catalytic beacon; it shows favorable peroxidase properties with a Michaelis contant (Km) of 0.072 mM for H2O2, which is capable of catalyzing H2O2 to induce an intense redox reaction, and causing a measurable electrochemical signal. To further enhance the strength of the signal response, a novel toehold-mediated DNA replacement strategy is employed. DNA strands with specific sequences are modified on electrodes and AuPt NPs, respectively. In the presence of miRNA-21, a cyclic substitution effect is subsequently activated via a specific toehold sequence and leads to a large accumulation of AuPt NPs on the electrodes. Subsequently, a strong signal depending on the amount of miRNA-21 is obtained after adding a small amount of H2O2. The analytical range of this determination method is from 0.1 pM to 1.0 nM, and the LOD is 84.1 fM. The spike recoveries for serum samples are 95.0 to 102.4% and the RSD values are 3.7 to 5.8%. The results suggests a promising application of the established method in clinical testing and disease diagnosis.


Subject(s)
Biosensing Techniques , MicroRNAs , MicroRNAs/chemistry , Hydrogen Peroxide/chemistry , Limit of Detection , Biosensing Techniques/methods , DNA/chemistry
8.
Arch Endocrinol Metab ; 67(3): 306-313, 2023 Mar 30.
Article in English | MEDLINE | ID: mdl-36468926

ABSTRACT

Objective: : Intrathyroidal injection using an insulin pen filled with a mixture of lidocaine and triamcinolone acetonide is a therapy for subacute thyroiditis (SAT) reported by us previously. We aimed to evaluate the clinical efficacy of ultrasound-guided intrathyroidal injection in the treatment of SAT. Subjects and methods: A total of 93 patients with SAT completed the study. All patients were evaluated via a history and clinical examination followed by thyroid function tests and ultrasonography of the thyroid. After ultrasound-guided intrathyroidal injection, the patients were followed up with respect to the injection frequency, treatment duration, and patient satisfaction. The visual numerical rating scale was used as a pain questionnaire for a given interval. Results: Thyroid pain instantly decreased to scores below 3.0 following the first injection. Sixty-three patients (67.74%) avoided relapse of thyroid pain within 3 injections, which occurred within only 3 days after the first injection. The pain in 27 patients (29.03%) disappeared completely after 4-6 injections. Only 3 patients (3.23%) were found to need more than 6 injections, with 10 cited as the maximum number of injections, the injection took only 17 days altogether. The mean treatment cycle of the intrathyroidal injection was 3.98 days. There were no other associated complications with the novel therapy except infrequent small subcutaneous hematomas, which could be prevented with skilled practice. The average patient satisfaction score was as high as 9.0. Conclusion: Intrathyroidal injection of lidocaine and triamcinolone acetonide using an insulin pen was found to be an advantageous and satisfactory treatment for SAT.


Subject(s)
Insulins , Thyroiditis, Subacute , Humans , Lidocaine/therapeutic use , Triamcinolone Acetonide/therapeutic use , Triamcinolone Acetonide/adverse effects , Anesthetics, Local/therapeutic use , Pain/chemically induced , Pain/drug therapy , Treatment Outcome , Insulins/therapeutic use
9.
Environ Sci Pollut Res Int ; 30(10): 27315-27327, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36378367

ABSTRACT

In this work, nitrogen-doped magnetic porous carbon material (N-MPC) was prepared through the high-temperature calcination of low-cost [Fe(CN)6]3--loaded anion-exchange resin, which was experimentally demonstrated to have significant adsorption performance for tetracycline (TC) in water. The N-MPC adsorbent with a large specific surface area (781.1 m2 g-1) was able to maintain excellent performance in a wide pH range from 4 to 10 or in high ionic strength solution. The adsorption of TC on N-MPC was found to be more consistent with the pseudo-second-order model and Langmuir adsorption model, and the maximum adsorption capacity (qm, cal) was calculated to be 603.4 mg g-1. As a recoverable magnetic adsorbent, the N-MPC remained a TC removal rate higher than 70% after four adsorption cycles. The adsorption mechanism was speculated on the basis of characterizations, where pore filling, hydrogen bonding interaction, and π-π electron donor-acceptor (EDA) interaction were crucial adsorption mechanisms. A variety of antibiotics were selected for adsorption, and excellent performance was found especially for TCs, indicating that the N-MPC can be used for the efficient removal of TCs from water.


Subject(s)
Carbon , Water Pollutants, Chemical , Carbon/chemistry , Water , Tetracyclines , Anion Exchange Resins , Nitrogen , Porosity , Anti-Bacterial Agents/chemistry , Tetracycline , Adsorption , Magnetic Phenomena , Water Pollutants, Chemical/analysis , Kinetics
10.
Mikrochim Acta ; 190(1): 41, 2022 12 31.
Article in English | MEDLINE | ID: mdl-36585965

ABSTRACT

A Cu2+-mediated turn-on fluorescence biosensor based on the DNA-templated green-emitting silver nanoclusters (DNA@g-AgNCs) was developed for label-free and sensitive detection of adenosine 5'-triphosphate (ATP). Cu2+ was able to quench the bright green fluorescence of DNA@g-AgNCs because of the coordination and photoinduced electron transfer between DNA@g-AgNCs and Cu2+. Therefore, a unique and effective fluorescence biosensor can be constructed with the formation of DNA@g-AgNCs/Cu2+/ATP ternary-competition system. With the introduction of ATP, the DNA@g-AgNCs/Cu2+ fluorescence sensing system will be disrupted and the fluorescence of DNA@g-AgNCs was recovered due to higher affinity of ATP towards Cu2+. On the basis of this feature, the DNA@g-AgNCs/Cu2+ fluorescence sensing system demonstrated quantitative determination of ATP in the range 0.05 - 3 µM and a detection limit of 16 nM. Moreover, the fluorescence sensing system was successfully applied to the quantitative determination of ATP in human urine and serum samples with recoveries ranging from 98.6 to 106.5%, showing great promise to provide  a label-free, cost-efficient, and rapid platform for ATP-related clinical disease diagnosis.


Subject(s)
Biosensing Techniques , Metal Nanoparticles , Humans , Silver , Adenosine Triphosphate , DNA
11.
Scand J Clin Lab Invest ; 82(7-8): 576-580, 2022.
Article in English | MEDLINE | ID: mdl-36336351

ABSTRACT

The objective of this study was to clinically and genetically characterize a pedigree with Liddle syndrome (LS). A LS pedigree comprising with one proband and seven family members was enrolled. The subjects' symptoms, laboratory results and genotypes were analyzed. Peripheral venous samples were collected from the subjects, and genomic DNA was extracted. DNA library construction and exome capture were performed on an Illumina HiSeq 4000 platform. The selected variant sites were validated using Sanger sequencing. The mutation effects were investigated using prediction tools. The proband and her paternal male family members had mild hypertension, hypokalemia and muscle weakness, including the absence of low renin and low aldosterone. Genetic analysis revealed that the proband carried a compound heterozygous mutation in SCNN1A, a novel heterozygous mutation, c.1130T > G (p.Ile377Ser) and a previously characterized polymorphism, c.1987A > G (p.Thr633Ala). The novel mutation site was inherited in an autosomal dominant manner and was predicted by in silico tools to exert a damaging effect. Alterations in the SCNN1A domain were also predicted by protein structure modeling. After six months of follow-up, treatment had significantly improved the patient's limb weakness and electrolyte levels. The novel mutation c.1130T > G of the SCNN1A gene was detected in the pedigree with LS. The clinical manifestations of the pedigree were described, which expand the phenotypic spectrum of LS. This result of this study also emphasizes the value of genetic testing for diagnosing LS.


Subject(s)
Hypertension , Hypokalemia , Liddle Syndrome , Female , Humans , Male , Hypokalemia/genetics , Epithelial Sodium Channels/genetics , Epithelial Sodium Channels/therapeutic use , Hypertension/genetics , Liddle Syndrome/diagnosis , Liddle Syndrome/drug therapy , Liddle Syndrome/genetics , Mutation , Pedigree
12.
Nat Cardiovasc Res ; 1(8): 732-747, 2022.
Article in English | MEDLINE | ID: mdl-35967457

ABSTRACT

Platelets have emerged as key inflammatory cells implicated in the pathology of sepsis, but their contributions to rapid clinical deterioration and dysregulated inflammation have not been defined. Here, we show that the incidence of thrombocytopathy and inflammatory cytokine release was significantly increased in patients with severe sepsis. Platelet proteomic analysis revealed significant upregulation of gasdermin D (GSDMD). Using platelet-specific Gsdmd-deficient mice, we demonstrated a requirement for GSDMD in triggering platelet pyroptosis in cecal ligation and puncture (CLP)-induced sepsis. GSDMD-dependent platelet pyroptosis was induced by high levels of S100A8/A9 targeting toll-like receptor 4 (TLR4). Pyroptotic platelet-derived oxidized mitochondrial DNA (ox-mtDNA) potentially promoted neutrophil extracellular trap (NET) formation, which contributed to platelet pyroptosis by releasing S100A8/A9, forming a positive feedback loop that led to the excessive release of inflammatory cytokines. Both pharmacological inhibition using Paquinimod and genetic ablation of the S100A8/A9-TLR4 signaling axis improved survival in mice with CLP-induced sepsis by suppressing platelet pyroptosis.

13.
Dent J (Basel) ; 10(8)2022 Aug 15.
Article in English | MEDLINE | ID: mdl-36005250

ABSTRACT

Objectives: To characterize the microflora profile of supragingival biofilm in patients with and without full-crown prostheses. Methods: Plaque samples of full-crown prostheses and teeth in patients with porcelain-fused-to-metal crowns, all-ceramic crowns, and no prostheses were collected (three patients per group), using 16S rRNA high-throughput sequencing technology to conduct DNA sequencing on the samples and using Qiime, R, and PICRUSt2 software to perform bioinformatics analyses and functional analyses on sequencing data. Results: In total, 110,209 valid sequences were obtained in the experiment, corresponding to 11 phyla and 120 genera. The predominant species shared by the three groups were phyla Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, and Proteobacteria and genera Rothia, Porphyromonas, Prevotella, Streptococcus, Veillonella, Leptotrichia, Neisseria, Citrobacter, and Pseudomonas. The species-difference analysis showed that genus Hameophilus significantly increased after the patient wore the dental prosthesis. Compared with the no-prosthesis samples, the functional analysis showed that cell motility increased in the samples from full-crown prostheses, while replication and repair, and translation decreased. Conclusions: This study reveals the changes in the oral microbial community of patients with full-crown prostheses, which could provide insights regarding the safety of materials for long-term use in the oral cavity.

14.
Front Immunol ; 13: 922868, 2022.
Article in English | MEDLINE | ID: mdl-35983051

ABSTRACT

Background: Kawasaki disease (KD) is an acute vasculitis that may result in permanent coronary artery damage with unknown etiology. Endothelial cell (EC) dysfunction and platelet hyperactivity are the hallmarks of KD. Platelets are involved in the development of endothelial dysfunction. MiR-223 transferred by platelet microparticles (PMPs) has been found to involve in the functional regulation of endothelial cells in sepsis. However, the role of platelet-derived miR-223 in endothelial dysfunction has not yet been investigated in KD. Objectives: We seek to investigate the role of platelet-derived miR-223 in endothelial dysfunction of KD vasculopathy. Methods and results: Forty-five acute KD patients and 45 matched controls were randomly recruited in the study. When co-cultured with human coronary artery endothelial cells (HCAECs), KD platelets with higher levels of miR-223 were incorporated into HCAECs, resulting in the horizontal transfer of miR-223. Using KD platelets, PMPs, and platelet-releasate from the same amount of blood co-cultured with HCAECs, we found the increased expression of miR-223 in HCAECs was primarily derived from KD platelets, rather than PMPs or free miRNAs from platelet- releasate. KD platelet-derived miR-223 attenuated TNF-α induced intercellular cell adhesion molecule-1 (ICAM-1) expression in HCAECs. KD platelet-derived miR-223 also suppressed the monocyte adhesion to HCAECs. In vivo, platelet-specific miR-223 knockout (PF4-cre: miR-223flox/flox) C57BL/6 mice and miR-223flox/flox C57BL/6 mice were used. Using Lactobacillus casei cell wall extract (LCWE) to establish KD murine model, we showed that in LCWE-injected PF4-cre: miR-223flox/flox mice, deficiency of platelet-miR-223 exacerbates the medial thickening of the abdominal aorta, increased ICAM-1 expression with concomitant CD45+ inflammatory cells infiltration into the endothelium compared to LCWE-injected miR-223flox/flox mice. Conclusions: The horizontal transfer of platelet-derived miR-223 suppresses the expression of ICAM-1 in HCAECs, which at least in part attenuates leukocyte adhesion, thereby reducing endothelial damage in KD vasculitis.


Subject(s)
MicroRNAs , Mucocutaneous Lymph Node Syndrome , Animals , Case-Control Studies , Endothelial Cells/metabolism , Endothelium, Vascular/metabolism , Humans , Intercellular Adhesion Molecule-1/genetics , Intercellular Adhesion Molecule-1/metabolism , Mice , Mice, Inbred C57BL , MicroRNAs/metabolism , Monocytes/metabolism , Mucocutaneous Lymph Node Syndrome/complications , Tumor Necrosis Factor-alpha/metabolism
15.
J Fluoresc ; 32(6): 2189-2198, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36030478

ABSTRACT

The construction of a universal nanoplatform for sensitive detection of multiple targets of interest is of great importance in different research fields. Herein, by ingeniously integrating the target recognition sequences and G-rich sequences into a single-stranded multifunctional DNA probe and adopting Ti3C2 nanosheets as an efficient fluorescence quencher, a simple, low-cost and easy operation fluorescence sensing nanoplatform was proposed. Without an analytical target, the hydrogen bond and metal chelate interaction between the target recognition region of the DNA probe and Ti3C2 nanosheet induce the selective self-assembly of highly fluorescent thioflavin T (ThT)-intercalated DNA probe onto the surface of Ti3C2 nanosheets, resulting in dramatic decrease of fluorescence emitted by ThT-G-quadruplex. In the presence of a target, the target recognition region will selectively bind with the target and the constrained DNA probe is released from the Ti3C2 nanosheets surface, leading to enhanced fluorescence recovery of ThT-G-quadruplex. As a proof of concept, the sensitive and selective detection of p53 gene, Hg2+, and adenosine with the assistance of Ti3C2 nanosheets-based fluorescence sensing nanoplatform were successfully realized. Moreover, it is also applicable for the evaluation the level of these analytical targets in real samples. By simply switching the recognition sequences of DNA probe, the universal sensing strategy could also be applied for detecting many other types of targets. The simple and universal sensing nanoplatform is expected to promote wide applications in environment monitoring and bioanalysis.


Subject(s)
Biosensing Techniques , G-Quadruplexes , Mercury , Fluorescence , Fluorescent Dyes/chemistry , DNA Probes , Mercury/analysis , DNA, Single-Stranded , Adenosine , Biosensing Techniques/methods , Limit of Detection
16.
Anal Chem ; 94(18): 6711-6718, 2022 05 10.
Article in English | MEDLINE | ID: mdl-35486137

ABSTRACT

Alkaline phosphatase (ALP), an essential hydrolase with crucial roles in living organisms, has widely been regarded as a biomarker for various human diseases in clinical diagnoses. Herein, taking advantage of cobalt oxyhydroxide (CoOOH) nanoflakes and nonenzymatic cascade recycling amplification (CRA), a highly sensitive and label-free fluorescence biosensing strategy for the determination of ALP activity is introduced. In our design, ALP can promote the dephosphorylation of l-ascorbic acid 2-phosphate (AAP) to reduce ascorbic acid (AA), which is then able to decompose CoOOH in a nucleic acids@CoOOH nanocomplex into Co2+ cofactors. Further, enzyme-free CRA was rapidly initiated by integrating DNAzyme recycling amplification and catalytic hairpin assembly, resulting in the generation of an abundance of G-quadruplex structure-contained DNA duplexes. In the presence of thioflavin T (ThT), analytical target ALP was converted in an amplified and activatable fluorescence signal. The experimental results show that this method can be applied for the quantitative analysis of ALP activity with a low detection limit of 0.027 mU/mL. Moreover, this developed biosensing approach exhibits excellent specificity, and the evaluation of ALP activity in the complex human serum samples was successfully realized, indicating that it can afford a reliable, robust, and cost-effective nanoplatform for an ALP-based clinical diagnosis and for biomedical research.


Subject(s)
Biosensing Techniques , DNA, Catalytic , Alkaline Phosphatase/analysis , Biosensing Techniques/methods , Cobalt , Fluorescent Dyes , Humans , Limit of Detection , Oxidation-Reduction , Oxides
17.
Scand J Clin Lab Invest ; 81(8): 629-633, 2021 12.
Article in English | MEDLINE | ID: mdl-34657521

ABSTRACT

Mutations in the SLC12A3 gene have been reported to cause Gitelman syndrome (GS). This study aimed to investigate the genetic mutations and clinical features of patients with GS. Four pedigrees (4 GS patients and 14 family members) were enrolled. The symptoms, laboratory results, management, and genotypes were analyzed. Genomic DNA was screened for gene variations using Sanger sequencing. DNA sequences were compared with reference sequences. The effects of the mutations were predicted using prediction tools (Mutation Taster, PolyPhen-2, SIFT, and PROVEAN). Genetic analysis revealed six genetic variants of SLC12A3, including three novel heterozygous mutations (c.2T > C, c.1609C > T, c.3055G > A) and three previously characterized mutations (c.1456G > A, c.2542G > A, c.1077C > G). These mutations were predicted to exert a damaging effect based on predictive in silico tools. GS patients had low blood pressure and low levels of serum K+, serum Mg2+, and 24-h urinary Ca2+ but high levels of 24-h urinary K+. These clinical manifestations and genotypes were consistent with the diagnostic criteria of GS. The study described the phenotypes and genotypes of 4 pedigrees involving GS patients, demonstrating the importance of SLC12A3 gene screening for GS.


Subject(s)
Gitelman Syndrome , Gitelman Syndrome/diagnosis , Gitelman Syndrome/genetics , Humans , Mutation , Pedigree , Phenotype , Solute Carrier Family 12, Member 3/genetics
18.
Mikrochim Acta ; 188(11): 385, 2021 10 18.
Article in English | MEDLINE | ID: mdl-34664146

ABSTRACT

A novel ratiometric fluorescence strategy for detection of acetylcholestinerase (AChE) is proposed based on carbon nitride quantum dots (g-CNQD) and the complex (PA) formed between phenylboronic acid (PBA) and alizarin red S (ARS). PA showed fluorescence at 598 nm and quenched the fluorescence of g-CNQD at 438 nm. Through UV-visible absorption, fluorescence, and fluorescence lifetime measurements, the quenching effect was demonstrated as inner filter effect (IFE). When Cu2+ was added, the coordination of ARS and Cu2+ decreased the fluorescence of PA at 598 nm and recovered that of g-CNQD at 438 nm. In the presence of AChE it catalyzed the hydrolysis of acetylthiocholine (ATCh) to produce thiocholine (TCh) which competed with ARS for binding to Cu2+; thus, the fluorescence at 598 nm increased and that at 438 nm decreased again. Under the mediation of Cu2+, the fluorescence ratio F598/F438 of PA-CNQD probe had good linear relationship with AChE concentration in the range 0.5-15 mU/mL with a detection limit of 0.36 mU/mL. The method was successfully applied to the determination of AChE in human serum and the screening of inhibitors.


Subject(s)
Acetylcholinesterase/blood , Biosensing Techniques/methods , Copper/chemistry , Fluorescence , Humans
19.
Mikrochim Acta ; 188(8): 264, 2021 07 21.
Article in English | MEDLINE | ID: mdl-34287718

ABSTRACT

A sensitive electrochemical strategy was established for kanamycin determination. A specific aptamer was modified on the electrode as the probe, followed by a cyclic hybridization chain reaction (HCR) with methylene blue, causing an increasing signal response. In the presence of kanamycin, it can initiatively convolve the aptamer and prevent further DNA assembling, resulting in a signal distinction sensitive to the target amount. However, the signal reproducibility is low. To improve the precision, the HCR procedure was investigated. The results demonstrated that the optimal amount of assembled DNA is 12-fold to that of aptamer. This amount was then controlled in further assays. Admittedly, controlled DNA assembling commonly indicates a limited signal amplification. To further enhance the sensitivity, a nanocomposite based on MoS2 and AuNPs was modified on the electrode. The results of the assay proved that the signal distinction sensitive to target amount increased by 50%. A linearity range is obtained from 0.01 nM to 1.0 µM of kanamycin, and the LOD is 8.4 pM. Subsequently, this strategy was employed to detect kanamycin in chicken liver and milk sample; the recovery results suggest that it possess a satisfactory application prospect in analysis of agricultural products.


Subject(s)
Disulfides/chemistry , Food Contamination/analysis , Gold/chemistry , Kanamycin/analysis , Metal Nanoparticles/chemistry , Molybdenum/chemistry , Nanocomposites/chemistry , Animals , Aptamers, Nucleotide/chemistry , Biosensing Techniques , Cattle , Chickens , DNA/chemistry , Electrochemical Techniques , Electrodes , Liver , Methylene Blue/chemistry , Milk , Nucleic Acid Amplification Techniques , Nucleic Acid Hybridization , Reproducibility of Results , Spectrometry, Fluorescence , Surface Properties
20.
Talanta ; 229: 122271, 2021 Jul 01.
Article in English | MEDLINE | ID: mdl-33838773

ABSTRACT

Hydrogen sulfide is typical metabolic marker and environmental pollutant which is worthwhile to determine. Herein, a low background and high sensitivity fluorescent strategy based on double modifications of metal organic framework material CAU-10-NH2 is proposed for the determination of hydrogen sulfide. Firstly, a functional monomer 3,5-diaminobenzoic acid is employed to modify on the CAU-10-NH2, the product CAU-10-NH-dAba has strong fluorescent performance at 412 nm under an excitation wavelength of 320 nm. Subsequently, it is further modified by the azide group to form CAU-10-NH-dAba-N3. This azidation inhibits the fluorescent signal. However, in the presence of hydrogen sulfide, the azide group is specifically reduced to amidogen, and results in the recovery of the fluorescence. The CAU-10-NH-DABA-N3 was characterized by solid state NMR, XPS, fluorescence, IR, XRD, SEM and specific surface area. After the optimization of pH value, temperature and interaction time, the detection results of hydrogen sulfide demonstrate the linear range of this strategy is from 20 to 140 nM with a detection limit of 1.51 nM, which is significantly better than that of the CAU-10-NH2 merely modified by 3,5-dinitrobenzoic acid. Meanwhile, the satisfactory assay results of hydrogen sulfide in serum sample and Pearl river water suggest a potential application prospect of this strategy in clinical diagnosis and environment monitoring.

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