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Through electrochemical polymerization using L-glutamic acid (L-Glu) as a template and 4,6-diaminoresorcinol as a functional monomer, an enzyme-free molecularly imprinted polymer (MIP) based L-Glu sensor with multi-walled carbon nanotubes (MWCNTs) decorated on a glassy carbon electrode (GCE), namely G-MIP/MWCNTs/GCE, was developed in this work. The reaction conditions were optimized as follows: electrochemical polymerization of 23 cycles, pH of 3.0, molar ratio of template/monomer of 1 : 4, volume ratio of elution reagents of acetonitrile/formic acid of 1 : 1, and elution time of 2 min. The prepared materials and molecularly imprinted polymer were characterized by using scanning electron microscopy (SEM) and transmission electron microscopy (TEM) as well as electrochemical methods. The electrochemical properties of different electrodes were investigated via differential pulse voltammetry (DPV), showing that the electrode of G-MIP/MWCNTs/GCE exhibited excellent catalytic oxidation activity towards L-Glu. A good linear relationship between peak-currents and L-Glu concentrations in a range from 1.00 × 10-8 to 1.00 × 10-5 mol L-1 was observed, with a detection limit of 5.13 × 10-9 mol L-1 (S/N = 3). The imprinted sensor possesses excellent selectivity, high sensitivity, and good stability, which have been successfully applied for the detection of L-Glu in pig serum samples with a recovery rate of 97.4-105.5%, being comparable to commercial high-performance liquid chromatography, demonstrating a simple, rapid, and accurate way for the determination of L-Glu in the fields of animal nutrition and biomedical engineering.
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Impresión Molecular , Nanotubos de Carbono , Porcinos , Animales , Polímeros Impresos Molecularmente , Nanotubos de Carbono/química , Ácido Glutámico , Impresión Molecular/métodos , ElectrodosRESUMEN
BACKGROUND: Gastric cancer is the fourth leading cause of cancer-related deaths worldwide. And patient outcomes are poor due to tumor relapse and metastasis. To develop new therapeutic strategies, it is of great importance to explore the mechanism underlying the progression of gastric cancer. METHODS: Primary gastric cancer samples with lymph node metastases (LNM) and without LNM were subjected to mRNA microarray assay. The differentially expressed genes were confirmed by RT-qPCR. HRCT1 protein expression was further detected using an immunohistochemistry (IHC) assay. In vitro and in vivo assays were performed to investigate the role of HRCT1 in tumor invasion, metastasis, and proliferation. The expressions of the downstream target genes of HRCT1 were detected by microarray, RT-qPCR and Western blot assays. Dual-luciferase reporter and Western blot assays were carried out to identify miRNAs target to HRCT1. RESULTS: HRCT1 was upregulated in gastric cancer, and high expression of HRCT1 was associated with poor overall survival (OS) and disease-free survival (DFS). Moreover, HRCT1protein expression was an independent predictor for poor OS and DFS. HRCT1 could promote gastric cancer cells' migration, invasion, and proliferation in vitro as well as tumor metastasis and growth in vivo. Notably, our data showed that HRCT1 promoted gastric cancer progression by activating the ERBB2-MAPK signaling pathway. At least partially, the expression of HRCT1 could be negatively regulated by miR-124-3p. CONCLUSIONS: The upregulated expression of HRCT1 predicts poor survival for patients with gastric cancer. HRCT1 promotes tumor progression by activating the ERBB2-MAPK pathway. HRCT1, negatively regulated by miR-124-3p, may be a potential therapeutic target for patients with gastric cancer.
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MicroARNs , Neoplasias Gástricas , Humanos , Neoplasias Gástricas/patología , Proliferación Celular/genética , Línea Celular Tumoral , MicroARNs/genética , Transducción de Señal , Metástasis Linfática , Regulación Neoplásica de la Expresión Génica , Movimiento Celular/genética , Receptor ErbB-2/metabolismoRESUMEN
BACKGROUND: To compare measurement properties of the utility scores derived from various country-specific value sets of EQ-5D-5L (5L) and EORTC QLU-C10D (10D) in gastric cancer patient. METHODS: The study used cross-sectional data of 243 Chinese gastric cancer patients who completed both 5L and EORTC QLQ-C30. Utility score of QLU-C10D is generated from all the available QLU-C10D value sets currently; the score of 5L is derived from the corresponding 5L value sets for the countries with both the 5L and QLU-C10D value sets and the Chinese 5L value set. Convergent validity was evaluated by testing their correlations with the VAS score. Known-group validity was assessed by comparing the utility scores the patients with different severities. Their relative efficiency (RE) was also compared. RESULTS: Correlation coefficient of 5L and QLU-C10D utility scores with VAS ranged from 0.54 to 0.59, and 0.55 to 0.63, respectively. Both the utility scores were in general able to discriminate the patients with different severities; and 5L utility score had higher RE in the majority of known-groups. CONCLUSION: EQ-5D-5L and QLU-C10D utility scores were different and, thus, non-swappable. They possess similar convergent validity and known-group validity; while EQ-5D-5L scores may have better discriminative power.
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Calidad de Vida , Neoplasias Gástricas , Humanos , Estudios Transversales , Encuestas y CuestionariosRESUMEN
Bladder cancer (BC) is the ninth most common lethal malignancy worldwide. Great efforts have been devoted to clarify the pathogenesis of BC, but the underlying molecular mechanisms remain unclear. To screen for the genes associated with the progression and carcinogenesis of BC, three datasets were obtained from the Gene Expression Omnibus. A total of 37 tumor and 16 non-cancerous samples were analyzed to identify differentially expressed genes (DEGs). Subsequently, 141 genes were identified, including 55 upregulated and 86 downregulated genes. The protein-protein interaction network was established using the Search Tool for Retrieval of Interacting Genes database. Hub gene identification and module analysis were performed using Cytoscape software. Hierarchical clustering of hub genes was conducted using the University of California, Santa Cruz Cancer Genomics Browser. Among the hub genes, kinesin family member 11 (KIF11) was identified as one of the most significant prognostic biomarkers among all the candidates. The Kaplan Meier Plotter database was used for survival analysis of KIF11. The expression profile of KIF11 was analyzed using the ONCOMINE database. The expression levels of KIF11 in BC samples and bladder cells were measured using reverse transcription-quantitative pCR, immunohistochemistry and western blotting. In summary, KIF11 was significantly upregulated in BC and might act as a potential prognostic biomarker. The present identification of DEGs and hub genes in BC may provide novel insight for investigating the molecular mechanisms of BC.
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OBJECTIVES: In China, multiple approaches to calculating EQ-5D utilities are available, including the two EQ-5D-3L (3L2014 and 3L2018) scoring functions, the EQ-5D-5L (5L) scoring function, and the crosswalk function linking the 3L utilities and 5L health states. The study compared utilities derived from them in terms of agreement and discriminative power; and assessed whether the use of different approaches may affect QALY estimation in Chinese type 2 diabetes (T2D) patients. METHODS: Cross-sectional data of 289 T2D patients who self-completed both the 5L and 3L questions were used. Agreement were examined using intraclass correlation coefficient (ICC) and Bland-Altman plots. The ability of the EQ-5D utilities in differentiating the patients with and without clinical conditions was evaluated using F-statistics. Their influence on QALY estimation was assessed adopting mean absolute difference (MAD) in utility values between the patients. RESULTS: The ICC values were 0.881 (3L2014-3L2018), 0.958 (5L-c5L2014), and 0.806 (5L-c5L2018). The two 3L utilities and the three 5L utilities had poor agreement at the lower end of utility scale according to Bland-Altman plots. The 3L2018 utilities had lower F-statistics compared to the 3L2014 utilities; the two c5L utilities had larger or similar F-statistics compared to the 5L utilities. The mean MADs were 0.138 (5L), 0.116 (3L2014), 0.115 (c5L2014), 0.055 (c5L2018), and 0.055 (3L2018). CONCLUSION: The 3L2014 utilities is more discriminative than the 3L2018 utilities; and the two c5L utilities have no worse discriminative power compared with the 5L utilities. The choice of the approach to calculating the EQ-5D utilities is likely to affect QALY estimates.
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Diabetes Mellitus Tipo 2/epidemiología , Estado de Salud , Calidad de Vida/psicología , Pueblo Asiatico , China , Estudios Transversales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Encuestas y CuestionariosRESUMEN
BACKGROUND: Clinically, the median survival in patients with metastatic renal cell carcinoma (RCC) was only 6-12 months and a 5-year survival rate of less than 20%. Therefore, an in-depth study of the molecular mechanisms involved in RCC is of great significance for improving the survival of patients with advanced RCC. Acylglycerol kinase (AGK) is a newly discovered lipid kinase that has been reported to be a potent oncogene that may be involved in the regulation of malignant progression in a variety of tumours. However, the expression and biological characteristics of the AGK gene in RCC remain unclear. METHODS: AGK expression was quantified by quantitative real-time PCR, Western blotting and immunohistochemistry in RCC cell lines and paired patient tissues. Kaplan-Meier method and Cox proportional hazards models were used to evaluate the prognostic value of AGK in human RCC tissue samples. Chi-squared test was performed to analyse the correlation between AGK expression and the clinicopathological features. Stable overexpression and knockdown of AGK in RCC cells was constructed with lentivirus. The oncogenic effects of AGK in human RCC progression were investigated using assays of colony formation, anchorage-independent growth, EdU assay, cell cycle analysis, wound-healing, trans-well analysis and xenograft tumour model. GSEA and KEGG analysis were conducted to detect the potential pathway of AGK involved in RCC. These results were further confirmed using the luciferase reporter assays, immunofluorescence and in vivo experiments. RESULTS: AGK expression is significantly elevated in RCC and closely related to the malignant development and poor prognosis in RCC patients. By in vitro and in vivo experiments, AGK was shown to enhance the proliferation of RCC cells by promoting the transition from the G1 phase to the S phase in the cell cycle and to enhance the migration and invasion by promoting epithelial-mesenchymal transition. By activating the PI3K/AKT/GSK3ß signalling pathway in RCC, AGK can increase nuclear accumulation of ß-catenin, which further upregulated TCF/LEF transcription factor activity. CONCLUSIONS: AGK promotes the progression of RCC via activating the PI3K/AKT/GSK3ß signalling pathway and might be a potential target for the further research of RCC.
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Carcinoma de Células Renales/patología , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Neoplasias Renales/patología , Fosfatidilinositol 3-Quinasas/metabolismo , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Carcinoma de Células Renales/metabolismo , Línea Celular Tumoral , Progresión de la Enfermedad , Activación Enzimática , Femenino , Humanos , Neoplasias Renales/metabolismo , Masculino , Persona de Mediana Edad , Metástasis de la Neoplasia/patología , Transducción de SeñalRESUMEN
A facile method was developed for synthesis of boronic acid-functionalized silica nanocomposites (SiO2 -BA) by 'thiol-ene' click reaction, where silica nanoparticles were synthesized by using tetraethoxysilane (TEOS) and γ-mercaptopropyl trimethoxysilane (γ-MPTS) as precursors. The morphology and structure properties of the resultant SiO2 -BA were characterized by transmission electronic microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), and Brunner-Emmet-Teller measurements (BET). The adsorption behavior of the SiO2 -BA for glycoproteins was evaluated. Under the optimized conditions, the SiO2 -BA exhibited higher adsorption capacity towards glycoproteins (ovalbumin, OVA, 7.64 µmol/g) than non-glycoproteins (bovine serum albumin, BSA, 0.83 µmol/g). In addition, the practicality of the SiO2 -BA was further assessed by selective enrichment of glycoproteins from egg white samples.
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Ácidos Borónicos/química , Glicoproteínas/química , Nanocompuestos/química , Dióxido de Silicio/química , Adsorción , Clara de Huevo/química , Estructura Molecular , Tamaño de la Partícula , Dióxido de Silicio/síntesis química , Propiedades de SuperficieRESUMEN
Breast cancer is the most common female malignancy worldwide, however its molecular pathogenesis still needs in-depth investigation. Here we first revealed that the olfactory receptor family 2, subfamily T, member 6 (OR2T6) was significantly over-expressed in breast cancer tissues compared with normal breast tissues. OR2T6 expression was tightly correlated with higher TNM staging, positive lymph node metastasis, and associated with poorer patients' overall and disease-free survival. And OR2T6 enhanced the proliferation, invasion, and migration ability of breast cancer cell lines in vitro (MCF-7 and MDA-MD-231). Mechanically, it promoted the expression of mesenchymal markers (Vimentin, N-cadherin, and ß-catenin) while inhibited E-cadherin expression, suggesting that OR2T6 played a key role in the regulation of epithelial-mesenchymal transition (EMT) process. Moreover, the human gene expression microarray clarified that MAPK/ERK pathway could be initiated by OR2T6 at mRNA level, which was further confirmed at protein level by western blot analysis. Thus, we concluded that OR2T6, as a novel oncogene, contributed to the progression of breast carcinoma by the initiation of EMT and MAPK/ERK pathway.
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The protocadherin 7 is a member of the protocadherin family that expressed aberrantly in many types of human cancers. However, its expression, function, and underlying mechanisms are little known in gastric cancer. In this study, we detected protocadherin 7 expression in gastric cancer tissues and non-tumorous gastric mucosa tissues by real-time quantitative polymerase chain reaction and immunohistochemistry. The association of protocadherin 7 expression with the clinicopathological characteristics and the prognosis was subsequently analyzed. MTS ((3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)) and transwell assays were performed to assess the effect of protocadherin 7 on proliferation, migration, and invasion in gastric cancer cell lines. Moreover, real-time quantitative polymerase chain reaction and western blot were used to detect the expression of epithelial-mesenchymal transition markers. Protocadherin 7 expression was decreased gradiently from normal tissue to gastric cancer, especially in gastric cancer tissue with lymph node metastasis. Low expression of protocadherin 7 was significantly associated with Lauren's classification ( p = 0.0005), lymph node metastases ( p = 0.0002), and tumor node metastasis stage ( p = 0.0221), as well as poor prognosis ( p < 0.05). Furthermore, down-regulation of protocadherin 7 in gastric cancer cell lines significantly increased their migration and invasion abilities (both p < 0.05), while it had no influence on the gastric cancer cell proliferation ( p > 0.05). Additionally, our results demonstrated that E-cadherin expression was down-regulated in gastric cancer cells with protocadherin 7 depletion. Our data indicated that protocadherin 7 may play important roles in the invasion and metastasis of gastric cancer, and protocadherin 7 could suppress cell migration and invasion through E-cadherin inhibition. Protocadherin 7 can serve as a novel biomarker for diagnostic and prognosis in patients with gastric cancer.
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Cadherinas/fisiología , Movimiento Celular , Neoplasias Gástricas/patología , Adulto , Anciano , Antígenos CD , Línea Celular Tumoral , Proliferación Celular , Femenino , Humanos , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Pronóstico , ProtocadherinasRESUMEN
Gallbladder Cancer (GBC), characterized by invasive growth and infiltrative dissemination, is difficult to diagnose and has poor prognosis. Emerging evidence demonstrates that Lysine-Specific Demethylase 1 (LSD1) has important roles in carcinogenesis, proliferation and metastasis. We studied the roles and molecular mechanisms of LSD1 in GBC. We examined LSD1 expression in 109 paired samples of GBC and normal gallbladder tissues. We found GBC tissues had upregulated LSD1 compared with normal gallbladder tissues (P = 0.003), and its high expression was associated with tumor-node-metastasis stage (P < 0.0001), Nevin's stage (P = 0.0093) and distant metastases (P = 0.0070). We found positive correlations between LSD1 expression and other proteins: epithelial-mesenchymal transition markers, C-myc and cyclin-related proteins. Inhibiting LSD1 expression in vitro impaired the proliferation and invasiveness of GBC cells and also downregulated c-myc expression and consequently inhibited GBC cell proliferation. LSD1 overexpression promotes GBC development and may be a predictor for a worsened prognosis. LSD1 may be a novel therapeutic target and prognostic tool for gallbladder cancer.
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Neoplasias de la Vesícula Biliar/enzimología , Histona Demetilasas/genética , Histona Demetilasas/metabolismo , Animales , Carcinogénesis/genética , Carcinogénesis/metabolismo , Línea Celular Tumoral , Proliferación Celular/fisiología , Regulación hacia Abajo , Transición Epitelial-Mesenquimal , Neoplasias de la Vesícula Biliar/genética , Neoplasias de la Vesícula Biliar/patología , Técnicas de Silenciamiento del Gen , Xenoinjertos , Humanos , Masculino , Ratones , Ratones Desnudos , Valor Predictivo de las Pruebas , Pronóstico , Regulación hacia ArribaRESUMEN
OBJECTIVE: To investigate the relationship between gene mutations and protein expressions of PDGFR α and C-kit in gastrointestinal stromal tumors (GIST) and its significance in tumorigenesis. METHODS: Single strand conformation polymorphism-polymerase chain reaction (PCR-SSCP), immunohistochemistry and Western blot were used to detect the gene mutations in PDGFR α and C-kit and their protein expressions in 105 cases of GIST specimens. RESULTS: In 105 cases of GIST, PDGFR α gene mutation was found in 12 cases (11.4%), which was common in the stomach- derived spindle cell GIST. C-kit gene mutation was found in 58 cases (55.2%), which was common in the small intestine. Mutations of PDGFR α is in 12 cases of GIST were stronger than the C-kit mutations in GIST, normal gastrointestinal tissues and schwannomas. No significant correlation was found between mutations and C-kit protein expression (P>0.05), while the protein expression of PDGFR α was significantly correlated with mutations (P<0.0001). CONCLUSION: Mutations of PDGFR α and C-kit plays an important role in part of GIST tumorigenesis. Mutation sites were related with original sites and histological types. Most protein expressions were closely related to their gene mutations in GIST.
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AIM: To investigate the microRNA (miRNA) expression profile in gastrointestinal stromal tumor (GIST) tissues that could serve as a novel diagnostic biomarker for GIST detection. METHODS: We performed a quantitative real-time quantitative reverse transcriptase polymerase chain reaction assay to analyze the expression of 1888 miRNAs in a sample set that included 54 GIST tissue samples. RESULTS: We found that dysregulation of several miRNAs may be related to the malignant potential of GISTs. Six of these miRNAs, hsa-let-7c, miR-218, miR-488#, miR-4683, miR-34c-5p and miR-4773, were selected as the final list of biomarkers to separate the malignant GISTs (M group) from the benign GISTs (B group). In addition, MiR-29b-2#, hsa-let-7c, miR-891b, miR-218, miR-204, miR-204-3p, miR-628-5p, miR-744, miR-29c#, miR-625 and miR-196a were used to distinguish between the borderline (BO group) and M groups. There were 11 common miRNAs selected to separate the benign and borderline (BB) group from the M group, including hsa-let-7c, miR-218, miR-628-5p, miR-204-3p, miR-204, miR-891b, miR-488#, miR-145, miR-891a, miR-34c-5p and miR-196a. CONCLUSION: The identified miRNAs appear to be novel biomarkers to distinguish malignant from benign GISTs, which may be helpful to understand the mechanisms of GIST oncogenesis and progression, and to further elucidate the characteristics of GIST subtypes.
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Biomarcadores de Tumor/genética , Neoplasias Gastrointestinales/genética , Tumores del Estroma Gastrointestinal/genética , Perfilación de la Expresión Génica/métodos , Secuenciación de Nucleótidos de Alto Rendimiento , MicroARNs/genética , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Adulto , Anciano , Femenino , Neoplasias Gastrointestinales/patología , Humanos , Masculino , Persona de Mediana Edad , Valor Predictivo de las PruebasRESUMEN
UNLABELLED: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide. Despite significantly improved diagnosis and treatment in recent years, the long-term therapeutic effect is compromised by the frequent recurrence and metastasis, of which the molecular mechanisms are not fully understood. Our initial studies in established HCC cell lines with different metastatic capabilities indicated a correlation of metastasis with the resistance to apoptosis and therefore the ability to survive in stressed conditions. Subsequent investigation revealed that increased expression of X-linked inhibitor-of-apoptosis protein (XIAP) was correlated with the resistance to apoptosis and enhanced invasiveness in vitro, which could contribute to increased metastatic foci in vivo. Furthermore, we found that nearly 90% of clinical samples from advanced HCC patients expressed high levels of XIAP. Patients with XIAP-positive tumors had a significantly increased risk of relapse, which resulted from metastasis after total liver resection and orthotopic liver transplantation. Indeed, XIAP expression could be an independent prognostic factor for predicting disease-free survival rate and overall survival rate of these patients. XIAP expression was also highly correlated with advanced cases that exceeded the Milan criteria and could be a prognostic factor for disease-free survival in these patients as well. CONCLUSION: Our studies have shown an important molecule in controlling HCC metastasis, defined a biomarker that can be used to predict HCC recurrence and patient survival after treatment, and suggest that XIAP can be a molecular target subject to intervention to reduce metastasis and recurrence.