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A unique combination of a specific nucleic acid restriction endonuclease (REase) and atom transfer radical polymerization (ATRP) signal amplification strategy was employed for the detection of T790M mutations prevalent in the adjuvant diagnosis of lung cancer. REase selectively recognizes and cleaves T790M mutation sites on double-stranded DNA formed by hybridization of a capture sequence and a target sequence. At the same time, the ATRP strategy resulted in the massive aggregation of upconverted nanoparticles (UCNPs), which significantly improved the sensitivity of the biosensor. In addition, the UCNPs have excellent optical properties and can eliminate the interference of autofluorescence in the samples, thus further improving the detection sensitivity. The proposed upconversion fluorescent biosensor is characterized by high specificity, high sensitivity, mild reaction conditions, fast response time, and a detection limit as low as 0.14 fM. The performance of the proposed biosensor is comparable to that of clinical PCR methods when applied to clinical samples. This work presents a new perspective for assisted diagnosis in the pre-intervention stage of tumor diagnostics in the early stage of precision oncology treatments.
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Técnicas Biossensoriais , Neoplasias Pulmonares , Humanos , Neoplasias Pulmonares/genética , Enzimas de Restrição do DNA , Receptores ErbB/genética , Polimerização , Clivagem do DNA , Limite de Detecção , Mutação , Medicina de Precisão , Inibidores de Proteínas Quinases , Técnicas Biossensoriais/métodosRESUMO
Exosome is an emerging tumor marker, whose concentration level can reflect the occurrence and development of tumors. The development of rapid and sensitive exosome detection platform is of great significance for early warning of cancer occurrence. Here, a strategy for electrochemical detection of A549-cell-derived exosomes was established based on DNA/ferrocene-modified single-walled carbon nanotube complex (DNA/SWCNT-Fc). DNA/SWCNT-Fc complexes function as a signal amplification platform to promote electron transfer between electrochemical signal molecules and electrodes, thereby improving sensitivity. At the same time, the exosomes can be attached to DNA/SWCNT-Fc nanocomposites via the established PO43--Ti4+-PO43- method. Moreover, the application of EGFR antibody, which can specifically capture A549 exosomes, could improve the accuracy of this sensing system. Under optimal experimental conditions, the biosensor showed good linear relationship between the peak current and the logarithm of exosomes concentration from 4.66 × 106 to 9.32 × 109 exosomes/mL with a detection limit of 9.38 × 104 exosomes/mL. Furthermore, this strategy provides high selectivity for exosomes of different cancer cells, which can be applied to the detection of exosomes in serum samples. Thus, owing to its advantages of high sensitivity and good selectivity, this method provides a diversified platform for exosomes identification and has great potential in early diagnosis and biomedical applications.
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Exossomos , Nanotubos de Carbono , Metalocenos , DNARESUMO
As a nonspecific phosphomonoesterase, alkaline phosphatase (ALP) plays a pivotal role in tissue mineralization and osteogenesis which is an important biomarker for the clinical diagnosis of bone and hepatobiliary diseases. Herein, we described a novel electrochemical method that used aminoferrocene (AFC) as an electroactive probe for the ALP activity detection. In the condition with imidazole and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC), the AFC probe could be directly labeled on single-stranded DNA (ssDNA) by one-step conjugation. Specifically, thiolated ssDNA at 3'-terminals was modified to the electrode surface through Au-S bond. In the condition without ALP, AFC could be labeled on ssDNA by conjugating with phosphate groups. In the presence of ALP, phosphate groups were catalyzed to be removed from the 5'-terminal of ssDNA. The AFC probe cannot be labeled on ssDNA. Thus, the electrochemical detection of ALP activity was achieved. Under optimal conditions, the strategy presented a good linear relationship between current intensity and ALP concentration in the range of 20 to 100 mU/mL with the limit of detection (LOD) of 1.48 mU/mL. More importantly, the approach rendered high selectivity and satisfactory applicability for ALP activity detection. In addition, this method has merits of ease of operation, low cost, and environmental friendliness. Thus, this strategy presents great potential for ALP activity detection in practical applications. An easy, sensitive and reliable strategy was developed for the detection of alkaline phosphatase activity via electrochemical "Signal off".
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Fosfatase Alcalina/análise , DNA de Cadeia Simples/análise , Eletroquímica/métodos , Enzimas/química , Compostos Ferrosos/química , Metalocenos/química , Fosfatase Alcalina/sangue , Animais , Técnicas Biossensoriais , Catálise , Bovinos , DNA de Cadeia Simples/sangue , Enzimas/sangue , Compostos Ferrosos/sangue , Glucose Oxidase/análise , Ouro/química , Humanos , Imidazóis/análise , Limite de Detecção , Metalocenos/sangue , Fosforilação , Reprodutibilidade dos Testes , Soro/química , Soroalbumina Bovina/análise , Enxofre/químicaRESUMO
Improving the sensitivity of detection is crucial to monitor biomarker, assess toxicity, and track therapeutic agent. Herein, a sensitivity-improved immunosensor is reported for the first time via functionalized graphene oxide (GO) and a "grafting-to" ring-opening polymerization (ROP) dual signal amplification strategy. Through the ROP reaction using 2-[(4-ferrocenylbutoxy)methyl] oxirane (FcEpo) as the monomer, lots of electroactive tags are linked in situ from multiple initiation sites on the GO surface modified with ethanol amine (GO-ETA), thereby achieving high sensitivity even in the case of trace amounts of tumor markers. The utmost important factor for achieving this high sensitivity is to select functionalized GO as the initiator that contains a large number of repeated hydroxyl functional groups so as to trigger additional ROP reaction. Under the optimal conditions, the high sensitivity and applicability is demonstrated by the use of GO-ETA-mediated ROP-based immunosensor to detect non-small cell lung cancer (NSCLC)-specific biomarker down to 72.58 ag/mL (equivalent to ~6 molecules in a 5 µL sample). Furthermore, the satisfactory results for the determination of biomarkers in clinical serum samples highlighted that this immunosensor holds a huge potential in practical clinical application. This work described an electrochemical immunosensor for ultrasensitive detection of CYFRA 21-1 via the functionalized graphene oxide (GO) and a "grafting-to" ring-opening polymerization (ROP) dual signal amplification strategy, which hold the merits of high sensitivity, applicability, selectivity, efficiency, easy operation and environmental friendliness.
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Antígenos de Neoplasias/sangue , Biomarcadores Tumorais/sangue , Grafite/química , Queratina-19/sangue , Fragmentos de Peptídeos/análise , Anticorpos Imobilizados/imunologia , Antígenos de Neoplasias/imunologia , Biomarcadores Tumorais/imunologia , Técnicas Eletroquímicas/métodos , Humanos , Imunoensaio/métodos , Queratina-19/imunologia , Limite de Detecção , Reprodutibilidade dos TestesRESUMO
CREPT (cell-cycle related and expression-elevated protein in tumor) was reported to be associated with growth of several human cancers; however, its clinical significance and regulatory mechanism still remain unclear in human gastric cancer. In the present study, we found CREPT was significantly increased in gastric cancer tissues compared to the matched adjacent normal tissues. CREPT silence inhibited the proliferation of gastric cancer cells through inducing G0/G1 phase cell cycle arrest, which was linked to the reduction of Cyclin D1 and Cyclin D-dependent kinase 4 (CDK4), and the elevation of p53 and p21. In addition, CREPT knockdown (KD) decreased migration of gastric cancer cells through up-regulating E-cadherin and down-regulating vimentin, N-cadherin and matrix metalloproteinase 1 (MMP-1) expressions. Further, CREPT KD induced apoptosis in gastric cancer cells, as evidenced by the increase of cleaved Caspase-3 and poly (ADP-ribose) polymerase (PARP). Intriguingly, suppressing p53 expressions significantly abolished CREPT silence-induced apoptosis, and reduction of cell viability. Moreover, CREPT KD caused reactive oxygen species (ROS) generation using discounted cash flow (DCF) analysis, which was reversed by ROS scavenger, N-acetyl-l-cysteine (NAC), pretreatment. Of note, NAC pretreatment abrogated apoptotic cell death in CREPT KD gastric cancer cells. In vivo, suppressing CREPT reduced the gastric tumor growth in gastric cancer xenograft models. Altogether, our results provided a novel insight into CREPT in regulating gastric cancer progression through apoptosis regulated by ROS/p53 pathways.
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Apoptose , Pontos de Checagem do Ciclo Celular , Proteínas de Ciclo Celular/metabolismo , Proteínas de Neoplasias/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Proteína Supressora de Tumor p53/metabolismo , Proteínas de Ciclo Celular/antagonistas & inibidores , Proteínas de Ciclo Celular/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Humanos , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/genética , Transdução de SinaisRESUMO
Immunotherapy has been used in esophageal cancer (EC), but the causal relationship between EC and immune cells is not clear. Although the cellular phenotype has been reported as a biomarker for immunotherapy, the biomarker studies for immunotherapy in EC still face great challenges. Comprehensive 2-sample Mendelian randomization (MR) analysis was performed to determine the causal association between immune cell signatures and EC in this study. Based on publicly available genetic data, we explored causal associations between 731 immune cell signatures and EC risk. EC had no statistically significant effect on immunophenotypes. Nine immunophenotype types were positively associated with the risk of EC: CD20-%B cell, CD20% lymphocytes, CD25 on IgD- CD27-, CD25 on IgD+ CD24+, CD27 on IgD+ CD24+, CD28+ CD45RA- CD8br AC, CD3 on TD CD8br, IgD-CD38dim%B cells, and Mo MDSC AC. In addition, a total of 15 immunophenotypes were identified as causally associated with EC. IgD+ CD38- %B cell, IgD- CD24- %lymphocyte, CD19 on IgD- CD38dim, CD20 on IgD+ CD24+, CD62L-myeloid DC AC, CD4+ AC, Lymphocyte %leukocyte, CD3 on HLA-DR+ T cell, CD3 on CD45RA- CD4+, HVEM on naive CD4+ AC, HVEM on CD45RA- CD4+, CD4 on TD CD4+, CD4 on CD4 Treg, and CD4 on CD39+ resting Treg, and CD4 on activated & secreting Treg. Our study has demonstrated the close connection between immune cells and EC by genetic means, thus providing guidance for future clinical research.
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Neoplasias Esofágicas , Imunofenotipagem , Análise da Randomização Mendeliana , Humanos , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/imunologia , Povo Asiático/genética , Ásia Oriental , População do Leste AsiáticoRESUMO
Objective: To delineate the immune landscape of ESCC patients mediated by aggrephagy through bioinformatics and identify prognostic cell cluster genes with causal attributes to esophageal cancer through Mendelian randomization. Methods: Quality control, dimension reduction, and annotation were performed on the ESCC single-cell dataset. NMF clustering of various cell subgroups was carried out based on the expression of AGG-related genes, and AGG-related genes in each cluster were identified. Pseudo-temporal analysis was used to observe changes in the expression of AGG-related genes in each cluster. Cell communication analysis was employed to observe interactions between cell subgroups. Changes in classification, metabolism, or KEGG pathways in related subgroups were observed based on different cell characteristics. The AGG cluster attributes of TCGA and GEO samples were assessed based on GSVA, and the prognosis of each cluster was observed. The immune treatment situation and the relationship between mutation level and prognosis of AGG cluster-related samples were observed through the TIDE database and microsatellite instability. Finally, the eQTL of genes in each prognostic AGG cluster was used as an instrumental variable, with esophageal cancer as the outcome factor. Through Mendelian randomization analysis, AGG cluster-related genes with a causal relationship to esophageal cancer were established. Results: Dimension reduction clustering of single-cell transcriptome data identified 19 different cell subgroups. After re-annotation of the 19 cell subgroups, it was found that the CAF cells, B cells, T cells, NK cells, etc., of ESCA patients were all elevated compared to the control group. CAF cells had a high degree of communication with most cells. There were significant differences in macrophage metabolism and B-cell-mediated signal transduction pathways in different AGG clusters. The TUBA1B+Mac-C0 cluster, along with other clusters, exhibits predictive prognostic and immunotherapeutic potential at the transcriptional level. Mendelian randomization analysis revealed a causal relationship between genes such as CTSZ, CTSC, DAD, COLEC12, ATOX1, within the AGG cluster, and the onset of esophageal cancer. Conclusion: Aggrephagy mediates and influences the alterations and interactions of various immune cells in patients with ESCC. We elucidate the roles of AGG-related clusters, such as TUBA1B+Mac-C0, VIM+CD8+T_cells-C0, UBB+Mac-C2, in mediating prognosis and immune therapy in ESCC patients. Genes causally associated with the occurrence of esophageal cancer are identified within the AGG cluster, including CTSZ, CTSC, DAD, COLEC12, ATOX1, etc., offering new evidence for clinical immune therapy. These findings underscore the significance of these gene clusters in influencing both prognosis and immune responses in the context of esophageal cancer, shedding light on potential therapeutic targets and prognostic markers.
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Background and objectives: Hashimoto's thyroiditis (HT), a chronic autoimmune disorder impacting thyroid function, is a growing public health concern. The relationship between Treg cells and HT has been extensively studied, with Treg cells considered crucial in suppressing HT progression. However, these studies have mainly been observational, limiting our understanding of Treg cells' impact on HT risk. Leveraging large datasets, we utilized Mendelian randomization (MR) analysis to examine the causal association between Treg cell biomarkers and HT, providing additional validation for these relationships. Methods: Comprehensive two-sample Mendelian randomization analysis was performed to determine the causal association between Treg cells signatures and HT in this study. Based on publicly available genetic data, we explored causal associations between 165 Treg cells signatures and HT risk. Results: The European cohort study has identified five Treg cell phenotypes that causally protect against HT risk. Resting Treg %CD4 (OR = 0.975, 95% CI = 0.954~0.998, P = 0.030); CD4 on resting Treg (OR = 0.938, 95% CI = 0.882~0.997, P = 0.041; CD28- CD8dim %CD8dim (OR = 0.983, 95% CI = 0.969~0.998, P = 0.030); CD25 on CD39+ resting Treg (OR = 0.926, 95% CI = 0.864~0.991, P = 0.026); 5) CD28 on activated & secreting Treg (OR = 0.969, 95% CI = 0.942~0.996, P = 0.025). The Asian cohort study has identified four Treg cell phenotypes negatively correlated with the risk of HT. CD25hi %T cell (OR = 0.635, 95% CI = 0.473~852, P = 0.002); CD4 Treg %CD4 (OR = 0.829, 95% CI = 0.687~1.000, P = 0.050); CD127-CD8br %T cell (OR = 0.463, 95% CI =0.311~0.687, P< 0.001); CD3 on resting Treg (OR = 0.786, 95% CI = 0.621~0.994, P = 0.044). Conclusion: Our study has demonstrated the close connection between Treg cells and HT by genetic means, thus providing foundational basis for future research.
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Doença de Hashimoto , Linfócitos T Reguladores , Humanos , Fatores de Proteção , Antígenos CD28 , Estudos de Coortes , Análise da Randomização Mendeliana , Doença de Hashimoto/genéticaRESUMO
Down-regulator of transcription 1 (DR1) is considered as a biomarker of hashimoto's thyroiditis (HT), which is a risk factor for thyroid cancer. Here, a label-free electrochemical biosensor for DR1 detection was constructed based on polyamidoamine (PAMAM) polymer and the nanocomposite (WO3@AuNPs) composed of tungsten trioxide (WO3) and gold nanoparticles (AuNPs). WO3@AuNPs was obtained by combining monolayer WO3 nanosheets, which has high conductivity, and AuNPs. The modification of WO3@AuNPs can not only increase the conductivity of the electrode but also provide more active sites for signaling units, thus greatly improve the sensitivity of the sensor. The polymer PAMAM is biocompatible and non-immunogenic, and its end functional group can bind to the target molecules, providing them with more binding sites and thus improving the sensitivity of the sensor. Under optimal conditions, the label-free biosensor showed a good linear relationship between the logarithm of DR1 concentration and the impedance in the range of 10â fg â mL-1 to 100â ng â mL-1, with a detection limit as low as 0.3â fg â mL-1. Besides, this label-free electrochemical platform exhibited satisfactory selectivity and anti-interference capability in human serum samples. Therefore, this method has considerable potential in clinical detection of DR1.
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Técnicas Biossensoriais , Técnicas Eletroquímicas , Ouro , Nanopartículas Metálicas , Óxidos , Tungstênio , Ouro/química , Nanopartículas Metálicas/química , Técnicas Eletroquímicas/métodos , Tungstênio/química , Humanos , Óxidos/química , Técnicas Biossensoriais/métodos , Poliaminas/química , Limite de DetecçãoRESUMO
BACKGROUND: Esophageal carcinoma (EC) is one of the most prevalent cancers in human populations worldwide. Baitouweng decoction is one of the most important Chinese medicine formulas, with the potential to treat cancer. AIM: To investigate the role and mechanism of Baitouweng decoction on EC cells. METHODS: Differentially expressed genes (DEGs) in EC tissues and normal tissues were screened by the cDNA microarray technique and by bioinformatics methods. The target genes of microRNAs were predicted based on the TargetScan database and verified by dual luciferase gene reporter assay. We used Baitouweng decoction to intervene EC cells, and detected the activity of EC9706 and KYSE150 cells by the MTT method. Cell cycle and apoptosis were measured by flow cytometry. The expression of BUB1 mRNA and miR-495-3p was measured by qRT-PCR. The protein levels of BUB1, STAT3, p-STAT3, CCNB1, CDK1, Bax, Caspase3, and Caspase9 were measured by Western blot analysis. The migration and invasion abilities of the cells were measured by wound-healing assay and Transwell invasion assay, respectively. RESULTS: DEGs identified are involved in biological processes, signaling pathways, and network construction, which are mainly related to mitosis. BUB1 was the key hub gene, and it is also a target gene of miR-495-3p. Baitouweng decoction could upregulate miR-495-3p and inhibit BUB1 expression. In vitro experiments showed that Baitouweng decoction significantly inhibited the migration and invasion of EC cells and induced apoptosis and G2/M phase arrest. After treatment with Baitouweng decoction, the expression of Bax, Caspase 3, and Caspase 9 in EC cells increased significantly, while the expression of BUB1, CCNB1, and CDK1 decreased significantly. Moreover, the STAT3 signaling pathway may play an important role in this process. CONCLUSION: Baitouweng decoction has a significant inhibitory effect on EC cell growth. BUB1 is a potential therapeutic target for EC. Further analysis showed that Baitouweng decoction may inhibit the growth of EC cells by upregulating miR-495-3p targeting the BUB1-mediated STAT3 signal pathway.
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Alkaline phosphatase (ALP) is a significant hydrolase enzyme found in living organisms, and the dysregulation of its physiological activity has been correlated with a variety of diseases. Exploring the activity of ALP has important implications for biomedical research and clinical diagnosis. Accordingly, we have developed a novel, highly sensitive electrochemical biosensor for the analysis of ALP. Based on photoinduced atom transfer radical polymerisation (photoATRP), this strategy combined a fabricated biosensor with hydrolysate produced by the hydrolysis of O-phosphoethanolamine by ALP. Furthermore, for signal amplification, photoATRP synthesises uses polymers with plentiful binding sites for ferrocenylmethyl methacrylate, and by using a photoredox catalyst under blue light irradiation to perform this without the need for copper complexes, it is beneficial for environmental protection compared to traditional atom transfer radical polymerisation (ATRP). The biosensor had a linear range of 10-150 mU·mL-1, with R2 = 0.998, and detection limits as low as 2.12 mU·mL-1. Moreover, by exhibiting outstanding selectivity and interference resistance in human serum samples, this sensor has great potential for practical applications.
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Fosfatase Alcalina , Técnicas Biossensoriais , Humanos , Polimerização , Bioensaio , Catálise , Limite de Detecção , Técnicas EletroquímicasRESUMO
Pancreatic cancer (PAC) remains one of the most lethal malignant neoplasms, which is diagnosed at an advanced stage and thus lose the chance for curative resection. Here, we further probed PAC with a comprehensive multi-omics approach. Using single-cell RNA sequencing, we provided an integrated analysis of ductal cell subpopulations over the Leiden algorithm to identify two mian subcluster: S100A6 + cells and FXYD2 + cells. The gene set enrichment analysis results show that the two subtypes focused on different pathways related to tumor development. Furthermore, we integrated bulk and single-cell RNA sequencing datasets to generate and validate the prognostic signatures of the overall survival (OS) in PAC patients and S100A6 + cells were significantly enriched in high-risk groups which had a poor prognosis. Collectively, this research expands our understanding of ductal cell and provides a new reliable prognosis signature in PAC.
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Accurate and ultrasensitive detection of cytokeratin 19 fragment (CYFRA21-1) is of vital importance for screening and diagnosis of potential lung cancer patient. In this paper, surface-modified upconversion nanomaterials (UCNPs) capable of aggregation by atom transfer radical polymerization (ATRP) were used as luminescent materials for the first time to achieve signal-stable, low-biological background, and sensitive detection of CYFRA21-1. Upconversion nanomaterials (UCNPs) feature extremely low biological background signals and narrow emission peaks, making them ideal sensor luminescent materials. The combination of UCNPs and ATRP not only improves sensitivity, but also reduces biological background interference for detecting CYFRA21-1. The target CYFRA21-1 was captured by specific binding of the antigen and the antibody. Subsequently, the end of the sandwich structure with the initiator reacts with monomers modified on UCNPs. Then, massive UCNPs are aggregated by ATRP that amplify the detection signal exponentially. Under optimal conditions, a linear calibration plot of the logarithm of CYFRA21-1 concentration versus the upconversion fluorescence intensity was obtained in the range of 1 pg/mL to 100 µg/mL with a detection limit of 38.7 fg/mL. The proposed upconversion fluorescent platform can distinguish the analogues of the target with excellent selectivity. Besides, the precision and accuracy of the developed upconversion fluorescent platform were verified by clinical methods. As an enhanced upconversion fluorescent platform of CYFRA21-1, it is expected to be useful in screening potential patients with NSCLC and provides a promising solution for the high-performance detection of other tumor markers.
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Técnicas Biossensoriais , Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Nanopartículas , Humanos , Antígenos de Neoplasias , Queratina-19 , Neoplasias Pulmonares/diagnóstico , Técnicas Biossensoriais/métodos , Limite de Detecção , Nanopartículas/químicaRESUMO
Exosomes, as a biomarker with enhancing tumor invasion and spread, play an essential role for lung cancer diagnosis, therapy, and prognosis. In this work, a novel electrochemical sensor was fabricated for detecting exosomes secreted by lung cancer cells based on polysaccharide-initiated ring-opening polymerization (ROP) and click polymerization. First, MPA formed a self-assembled monolayer on the gold electrode surface, and then anti-EGFR was immobilized on the electrode surface by amide bond. Subsequently, a lot of phosphate groups were introduced by the specific recognition between anti-EGFR and exosomes, then sodium alginate grafted Glycidyl propargyl ether (SA-g-GPE) prepared via ROP was attached to the exosomes through PO43-Zr4+-COOH coordination bond. After that, click polymerization was initiated by alkyne groups on the SA-g-GPE polymerization chain to realize highly sensitive detection of A549 exosomes. Under the optimum conditions, the fabricated sensor showed a good linear relationship between the logarithm of exosomes concentration and peak current in the range of 5 × 103 - 5 × 109 particles/mL, and the limit of detection (LOD) was as low as 1.49 × 102 particles/mL. In addition, this method had the advantages of high specificity, anti-interference, high sensitivity, simplicity, rapidity and green economy, which proposed a novel avenue for the detection of exosomes, and also had potential applications in early cancer diagnosis and biomedicine.
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Técnicas Biossensoriais , Exossomos , Técnicas Eletroquímicas , Ouro , Limite de Detecção , Polimerização , PolissacarídeosRESUMO
PURPOSE: To study the effect of osthole extract on proliferation, migration, invasion, and apoptosis of human cervical carcinoma HeLa cells and investigate its underlying mechanism. METHODS: HeLa cells were exposed to osthole at various concentrations. Cell viability, migration, and invasion were detected by MTT assay, scratch wound-healing assay, and invasion assay, respectively. The proportion of cells undergoing apoptosis was analyzed by flow cytometry. Western blot and RT-qPCR were performed to determine changes in the expression of key factors in the Wnt/ß-catenin signaling pathway. RESULTS: The osthole extract effectively inhibited the proliferation, migration, and invasion potential of HeLa cells in a dose-dependent manner. The rate of apoptosis induction in HeLa cells treated with the osthole extract for 48 h was significantly higher than that of the untreated controls. Outcomes of the western blotting analysis and RT-qPCR showed that the expression of ß-catenin, c-Myc, cyclin D1, survivin, and MMP-9 was significantly inhibited. CONCLUSION: Osthole could significantly inhibit the malignant behavior of HeLa cells and induce cellular apoptosis. Inactivation of the Wnt/ß-catenin signaling pathway by osthole may be a mechanism to control cancer metastasis.
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As an important hydrolytic enzyme, abnormal activity of alkaline phosphatase (ALP) is closely associated with a variety of diseases. It has been identified as an important diagnostic indicator for clinical hepatobiliary and bone diseases. Herein, a novel electrochemical sensor based on signal amplification strategy through ring-opening polymerization (ROP) has been developed to assay of ALP activity. First of all, 3-mercaptopropanoic acid (MPA) was employed as a cross-linking agent to attach O-phosphoethanolamine to the electrode surface via amide bond. Then, ALP catalyzed the hydrolysis of phosphate monoester structures to hydroxyl groups, which could initiate ROP reaction. The polymer grafted on the electrode surface contains a large number of ferrocene electroactive molecules, which effectively increased the signal output of the electrochemical sensor and improved the sensitivity of ALP activity detection. Under optimum conditions, this electrochemical sensor rendered a satisfactory linear dependence over the range from 20 to 120 mU mL-1, with a low detection limit of 0.66 mU mL-1. Furthermore, this strategy presented satisfactory selectivity and interference resistance in human serum sample, and compared with clinical data, the relative error of the results obtained by this method was less than 5%. Thus, this method showed considerable potential for the detection of ALP activity in clinical application.
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Fosfatase Alcalina , Técnicas Biossensoriais , Bioensaio , Técnicas Eletroquímicas , Eletrodos , Humanos , Limite de Detecção , Polimerização , PolímerosRESUMO
To promote the development of Traditional Chinese Medicine (TCM), it is necessary to innovate the traditional prescription. It is feasible to use one or several components to substitute TCM, which can be regarded as a process of discarding the dregs and preserving the essential components. In this way, traditional prescription can be converted into various combinations of pharmacological ingredients deriving from several TCMs. Furthermore, some of pharmacological ingredients should be modified to increase their efficacy. It is practical to select the main structural unit with specific substituents having strong pharmacological activity. After the innovation mentioned above, the prescription will evolve into a variety of modified components having distinct pharmacological activity, and this is the novel integration of active ingredients.