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Disulfiram (DSF), an old anti-alcoholism drug, has emerged as a candidate for drug repurposing in oncology. In exploratory studies on its therapeutic effects, we unexpectedly discovered that DSF increased the phosphorylation of SRC, a proto-oncogene tyrosine-protein kinase elevated in 70% of pancreatic ductal adenocarcinoma (PDAC) cases. This serendipitous and novel finding led to our hypothesis for the current study which proposes DSF may synergize with SRC inhibitors in suppressing PDAC. Human PDAC PANC-1 and BXPC-3 cells were incubated with DSF chelated with copper (Cu2+), SRC inhibitors (PP2 and dasatinib), or transfected with lentiviral short hairpin RNA (shRNA), and their proliferation and apoptosis were analyzed. A xenograft model was employed to verify the in vitro results. The expression of key molecules was detected. DSF significantly inhibited cell proliferation and induced cell apoptosis by increasing the cleavage of poly ADP ribose polymerase (PARP), downregulating Bcl-2 and upregulating p27 in concentration- and time-dependent manners. DSF had little effect on signal transducer and activator of transcription 3 (STAT3) expression but inhibited its phosphorylation. DSF did not alter SRC expression but significantly increased its phosphorylation through upregulating actin filament associated protein 1 like 2 (AFAP1L2). DSF exhibited a synergistic effect, as analyzed by drug coefficient interactions, with either PP2, or dasatinib, or SRC depletion in suppressing PDAC cells in vitro and/or in vivo. The present results indicate DSF is a potential therapeutic drug, particularly when it is combined with SRC inhibitors, and warrant further studies on the pharmacological utility of DSF as a promising adjunct therapy for the treatment of PDAC.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Carcinoma Ductal Pancreático/tratamento farmacológico , Dissulfiram/farmacologia , Neoplasias Pancreáticas/tratamento farmacológico , Quinases da Família src/antagonistas & inibidores , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma Ductal Pancreático/patologia , Linhagem Celular Tumoral , Dasatinibe/farmacologia , Dasatinibe/uso terapêutico , Dissulfiram/uso terapêutico , Sinergismo Farmacológico , Humanos , Masculino , Camundongos , Neoplasias Pancreáticas/patologia , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , Pirimidinas/farmacologia , Pirimidinas/uso terapêutico , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
ß-Galactosidase (ß-Gal), as a glycoside hydrolase, is closely associated with cell senescence and primary ovarian cancer. However, there is still lack of facile and rapid sensing approach to monitor the ß-Gal activity. In this work, a label-free and convenient sensing strategy to detect ß-Gal activity has been proposed based on fluorescent graphene quantum dots (GQDs). In the presence of ß-Gal, 4-nitrophenyl-ß-D-galactopyranoside (NPGal) can be hydrolyzed into 4-nitrophenol (4-NP), which serves as a good quencher to quench the fluorescence of GQDs. The quenching mechanism is proven to be inner filter effect (IFE). Due to the specificity of the enzymatic reaction, this sensing method displays excellent selectivity and high sensitivity. A broad dynamic range from 20 to 200 U L-1 and a detection limit of 4.4 U L-1 for the ß-Gal assay are achieved. Compared with the previously reported methods, this sensing strategy only needs one fluorescent nanomaterial without any modification and avoids time-consuming handling steps. Therefore, the sensing strategy based on fluorescent GQDs offers great potential for the recognition of disease-correlated enzyme activity.
Assuntos
Técnicas Biossensoriais , Grafite , Pontos Quânticos , Espectrometria de Fluorescência , beta-GalactosidaseRESUMO
BACKGROUND: CHD is reported to be the primary cause of death in patients with NAFLD. Genetic susceptibility genes contribute to the developmental risk of NAFLD or CHD. Whether the genetic factors could affect the risk of CHD in NAFLD patients is not clear. The aim of this study was to investigate the association of PNPLA3 I148M and TM6SF2 E167K variants with the risk of CHD in NAFLD patients in Chinese Han population. PATIENTS AND METHODS: PNPLA3 I148M and TM6SF2 E167K variants were genotyped in a cohort of 189 patients with NAFLD and CHD, as well as 242 patients with NAFLD and 242 healthy controls by gene sequencing. Additionally, serum lipids profiles were determined by standard clinical laboratory methods. RESULTS: The minor allele frequency of PNPLA3 I148M and TM6SF2 E167K were 0.39 and 0.06 in this cohort, respectively. The distributions of PNPLA3 I148M genotypes and alleles were significant different in NAFLD group vs controls and in NAFLD+CHD group vs NAFLD group (all P < 0.05). NAFLD patients who carry the CG + GG genotype suffered the relative lower risk of CHD than CC genotype carriers (OR = 0.6, 95%CI: 0.40-0.90, P = 0.01). In addition, PNPLA3 I148M and TM6SF2 E167K possess the joint correlation with the decreased risk of CHD in NAFLD patients with the increased number of risk alleles. Besides, PNPLA3 I148M and TM6SF2 E167K variants associated with the decreased serum lipid levels in overall series. CONCLUSIONS: There was a joint protective correlation of PNPLA3 I148M and TM6SF2 E167K variants with the developmental risk of CHD in NAFLD patients. PNPLA3 I148M and TM6SF2 E167K variants might correlated with the decreased risk of CHD in NAFLD patients by associated with the reduced serum lipid levels.
Assuntos
Doença das Coronárias/genética , Lipase/genética , Proteínas de Membrana/genética , Hepatopatia Gordurosa não Alcoólica/genética , Alelos , Doença das Coronárias/sangue , Feminino , Frequência do Gene/genética , Predisposição Genética para Doença/genética , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/sangue , Polimorfismo de Nucleotídeo Único/genéticaRESUMO
Vinculin is a highly conserved protein involved in cell proliferation, migration, and adhesion. However, the effects of vinculin on gastric cancer (GC) remain unclear. Therefore, we aimed to explore the functional role of vinculin in GC, as well as its underlying mechanism. Expression of vinculin in patients with GC was analyzed by real-time polymerase chain reaction, Western blot analysis, and immunohistochemistry. Overall survival was evaluated by the Kaplan-Meier method with the log-rank test. The relationship between vinculin and clinicopathological characteristics of patients with GC was further identified. In addition, we assessed the expression of vinculin in GC cell lines. Besides, vinculin was suppressed or overexpressed by transfection with small interfering (si-vinculin) or pcDNA-vinculin and then cell viability, cell apoptosis, and/or migration was respectively examined by the 3-(4, 5-dimethylthiazole-2-yl)-2, 5-biphenyl tetrazolium bromide assay, flow cytometer, and scratch assay, respectively. Moreover, the cell cycle- and apoptosis-related proteins were detected by Western blot analysis. The expression of vinculin was significantly increased in the GC tissues and cells compared with the nontumor tissues or cells. Vinculin protein positive staining was mainly located in the cell membrane and cytoplasm. Moreover, vinculin was significantly associated with Tumor Node Metastasis (TNM) and poor differentiation. Patients with high vinculin levels had significantly worse overall survival than those with low levels. Suppression of vinculin significantly decreased cell viability and migration and promoted cell apoptosis. However, overexpression of vinculin statistically increased cell viability but had no effects on cell apoptosis. Vinculin promotes GC proliferation and migration and predicts poor prognosis in patients with GC.
Assuntos
Movimento Celular , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Vinculina/metabolismo , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Sobrevivência Celular/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Prognóstico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Neoplasias Gástricas/genética , Vinculina/genéticaRESUMO
Phosphoprotein enriched in astrocytes 15 (PEA15) plays an important role in controlling biological behaviors of cancer cells. In the present study, we demonstrated that PEA15 was overexpressed in gastric cancer tissues and associated with tumor staging, differentiation, pathological types and the prognosis of patients. Gastric cancer cells expressed variable levels of PEA15 and its biphosphorylation forms, pPEA15 (Ser104) and pPEA15 (Ser116). To gain insight into the functional role of PEA15, we generated cells stably depleted of PEA15 and resistant to cisplatin (CDDP) from human gastric cancer cells. PEA15 depletion inhibited cell proliferation by reducing cyclin D1 expression through the extracellular signalregulated kinase (ERK) pathway, resulting in cell cycle arrest at the G1 phase, and induced apoptosis by activating caspase8. PEA15 depletion also enhanced the inhibitory effect of CDDP that caused cell cycle arrest at the S phase and also enhanced the proapoptotic activity of CDDP in vitro and in animal models of tumorigenesis and therapeutic effects. PEA15 and its phosphorylated forms were overexpressed in CDDPresistant cells, which had higher levels of pAKT. Specific inhibition of AKT by MK2206 reduced the expression of pPEA15 at the Ser116 residue, resulting in sequential downregulation of pERK1/2, cyclin D1 and caspase8 activation. However, depletion of PEA15 had little effect on AKT expression or phosphorylation, or its downstream factors including p27, glycogen synthase kinase 3ß and caspase9, indicating that the regulatory effects between PEA15 and AKT were unidirectional. In summary, the results indicated that PEA15 expression was associated with clinicopathology and prognosis in gastric cancer and was regulated by AKT to participate in CDDP resistance, indicating that it may be a potential target for overcoming CDDP resistance in the treatment of gastric cancer.
Assuntos
Antineoplásicos/farmacologia , Cisplatino/farmacologia , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Fosfoproteínas/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Neoplasias Gástricas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Antineoplásicos/uso terapêutico , Apoptose , Proteínas Reguladoras de Apoptose , Linhagem Celular Tumoral , Cisplatino/uso terapêutico , Regulação para Baixo , Resistencia a Medicamentos Antineoplásicos , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Estimativa de Kaplan-Meier , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Fosfoproteínas/genética , Fosforilação , Prognóstico , RNA Interferente Pequeno/metabolismo , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/mortalidade , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Hepatic encephalopathy (HE) is a neuropsychiatric syndrome resulting from chronic or acute liver failure. Under the condition of HE, various factors such as reactive oxygen species, inflammatory factors, ammonia poisoning and amino acids alteration lead to changes of mitochondria. Selective depletion of damaged mitochondrion is essential for maintaining the morphology and function of mitochondria and cells. In this study, molecular biology analysis was used to analyze the mitochondrial morphology in the substantia nigra (SN) and anterior cerebral cortex (ACC) of the HE mice. The results revealed that the drp1, mfn1 and mfn2 increased in mRNA level of SN, which indicated the changes of mitochondrial morphology in HE mice. The drp1 and mfn2 genes were up-regulated, then, the Opa1 exhibited no significant change in the ACC of HE mice. Further study demonstrated that the mitochondrial autophagy related genes, pink1 and parkin, increased in SN, while the parkin reduced in ACC of HE mice. In addition, uncoupling protein (ucp2) increased in mRNA level of SN and ACC, and the ucp4 had no change or reduced in SN and ACC, respectively. These findings suggested that the mitochondrial dynamics is different in the SN and ACC of HE mice. Therefore, our results indicated that mitochondrial dynamics provided a potential treatment strategy for HE through the fission, fusion and autophagy of genes. Anat Rec, 302:1169-1177, 2019. © 2018 The Authors. The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.
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Córtex Cerebral/patologia , Encefalopatia Hepática/patologia , Mitocôndrias/patologia , Substância Negra/patologia , Animais , Autofagia , Córtex Cerebral/citologia , Modelos Animais de Doenças , Encefalopatia Hepática/induzido quimicamente , Humanos , Injeções Intraperitoneais , Masculino , Camundongos , Dinâmica Mitocondrial , Substância Negra/citologia , Tioacetamida/administração & dosagem , Tioacetamida/toxicidadeRESUMO
Gastric cancer (GC) is one of the leading malignancies worldwide and is also a leading cause of cancer-related mortality. Micro RNA (miRNA) is a group of short non-coding RNAs modulating gene expression through targeting the 3'UTR of genes. Peripheral blood exosome miRNAs are relatively stable and might be used as biomarkers for clinical diagnosis. In this study, we found a reduced level of miR-129-3p in GC tumor when compared with adjacent non-tumor tissues, and the peripheral blood exosome miR-129-3p level was also reduced when compared with healthy controls. Subsequently, we identified that miR-129-3p repressed the expression of SUMO-activating enzyme subunit 1 (SAE1) directly through targeting 3'UTR. miR-129-3p also inhibited the sumoylation modification of XRCC4 which disturbed the nuclear localization of XRCC4 and induced more DNA damage in GC cells. Furthermore, overexpression of miR-129-3p induced more cell apoptosis and inhibited GC cell proliferation, migration and invasion, indicating miR-129-3p is a powerful anti-tumor miRNA and has potential for GC treatment.
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MicroRNAs are important regulators during human growth and development. Emerging evidence indicates that microRNAs play important roles in colorectal cancer. The aim of this study is to reveal the biological function and direct target gene of miR-483 in colorectal cancer. The biological function of miR-483 on the proliferation and migration of colon cancer cells was then examined by Edu assay and transwell assay, respectively. Our findings revealed that miR-483 mimic could significantly inhibit cell proliferation and migration. The target gene of miR-483 was predicted by target scan software and identified by a dual fluorescence reporter system which showed that TRAF1 was a direct target gene of miR-483 in SW480 cell line. These data suggest that miR-483 is a colorectal cancer suppressor which could inhibit cell proliferation and migration, possibly via targeting TRAF1. The miR-483 could be a potential treatment target for colorectal cancer.
Assuntos
Neoplasias do Colo/metabolismo , MicroRNAs/metabolismo , Fator 1 Associado a Receptor de TNF/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Movimento Celular/fisiologia , Proliferação de Células/genética , Proliferação de Células/fisiologia , Neoplasias do Colo/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Reação em Cadeia da Polimerase em Tempo Real , Fator 1 Associado a Receptor de TNF/genéticaRESUMO
Activated leukocyte cell adhesion molecule (ALCAM/CD166) is a transmembrane glycoprotein that is involved in tumor progression and metastasis. In the present study, the expression and functional role of ALCAM in pancreatic cancer cells and pancreatic stellate cells (PSCs) was investigated. Tissue specimens were obtained from patients with pancreatic ductal adenocarcinoma (n=56) or chronic pancreatitis (CP; n=10), who underwent pancreatic resection, and from normal pancreatic tissue samples (n=10). Immunohistochemistry was used to analyze the localization and expression of ALCAM in pancreatic tissues. Subsequently, reverse transcriptionquantitative polymerase chain reaction and immunoblotting were applied to assess the expression of ALCAM in pancreatic cancer Panc1 and T3M4 cells, as well as in PSCs. An enzymelinked immunosorbent assay was used to measure ALCAM levels in cell culture medium stimulated by hypoxia, tumor necrosis factor (TNF)α and transforming growth factorß. Silencing of ALCAM was performed using ALCAM small interfering (si)RNA and immunocytochemistry was used to analyze the inhibition efficiency. An invasion assay and a cell interaction assay were performed to assess the invasive ability and cocultured adhesive potential of Panc1 and T3M4 cells, as well as PSCs. Histologically, ALCAM expression was generally weak or absent in pancreatic cancer cells, but was markedly upregulated in PSCs in pancreatic cancer tissues. ALCAM was highly expressed in PSCs from CP tissues and PSCs surrounding pancreatic intraepithelial neoplasias, as well as in pancreatic cancer cells. ALCAM mRNA was highly expressed in PSCs, with a low to moderate expression in T3M4 and Panc1 cells. Similar to the mRNA expression, immunoblotting demonstrated that ALCAM protein levels were high in PSCs and T3M4 cells, but low in Panc1 cells. The expression of TNFα increased, while hypoxia decreased the secretion of ALCAM in pancreatic cancer Panc1 and T3M4 cells, and also in PSCs. Silencing of ALCAM by siRNA revealed no significant alteration in the invasion of pancreatic cancer cells, however, it inhibited the invasive ability of PSCs, and decreased the interaction between Panc1 cells and PSCs. In conclusion, ALCAM is upregulated in PSCs of pancreatic cancer tissues, suggesting a potential role of ALCAM in regulating pancreatic cancer cellPSC interactions.
Assuntos
Molécula de Adesão de Leucócito Ativado/análise , Carcinoma Ductal Pancreático/patologia , Pâncreas/patologia , Neoplasias Pancreáticas/patologia , Células Estreladas do Pâncreas/patologia , Molécula de Adesão de Leucócito Ativado/genética , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/metabolismo , Comunicação Celular , Linhagem Celular , Regulação Neoplásica da Expressão Gênica , Humanos , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Pâncreas/citologia , Pâncreas/metabolismo , Neoplasias Pancreáticas/genética , Células Estreladas do Pâncreas/citologia , Células Estreladas do Pâncreas/metabolismo , Pancreatite/genética , Pancreatite/patologia , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Regulação para CimaRESUMO
5-Azacytidine is a well-known anticancer drug that is clinically used in the treatment of breast cancer, melanoma and colon cancer. It has been reported that 5-azacytidine suppresses the biological behavior of esophageal cancer cells. However, corresponding mechanisms remain unclear. In this study, using Transwell invasion and cell proliferation assays, we demonstrated that 5-azacytidine significantly inhibited the metastasis and proliferation of EC9706 cells, and upregulated the expression of cadherin 1 (CDH1) and SRY-box containing gene 17 (SOX17). Moreover, the inhibition of the metastasis of the 5-azacytidine-treated EC9706 cells was impaired following transfection with siRNA targeting CDH1 (CDH1 siRNA), and the inhibition of cell proliferation was attenuated following the downregulation of SOX17 by siRNA targeting SOX17 (SOX17 siRNA). Furthermore, 5-azacytidine remarkably reduced the CDH1 and SOX17 promoter methylation levels, suggesting that 5-azacytidine upregulates the expression of SOX17 and CDH1 by inhibiting the methylation of the SOX17 and CDH1 promoter. The findings of our study confirm that 5-azacytidine suppresses the proliferation and metastasis of EC9706 esophageal cancer cells by upregulating the expression of CDH1 and SOX17. The expression levels of CDH1 and SOX17 negatively correlate with the promoter methylation levels. CDH1 and SOX17 are potential indicators of the clinical application of 5-azacytidine.
Assuntos
Azacitidina/farmacologia , Caderinas/genética , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Fatores de Transcrição SOXF/genética , Regulação para Cima/efeitos dos fármacos , Antígenos CD , Caderinas/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Metilação de DNA/efeitos dos fármacos , Metilação de DNA/genética , Humanos , Metástase Neoplásica , Regiões Promotoras Genéticas/genética , Fatores de Transcrição SOXF/metabolismoRESUMO
BACKGROUND: Neuropilin-1 (NRP-1) is a transmembrane glycoprotein participating in the growth and metastasis of cancer cells as multifunctional co-receptors by interacting with the signaling pathways. However, its role in gastric cancer has not yet been clarified. This study aims to investigate whether NRP-1 expression is associated with the clinicopathology of gastric cancer, and involved in the growth and metastasis of gastric cancer cells. METHODS: NRP-1 expression in clinical gastric cancer specimens was examined by immunohistochemistry and its association with clinicopathology analyzed. The expression of NRP-1 in a panel of human gastric cancer cells was examined by real-time RT-PCR and immunoblotting. Stable transfectants depleted of NRP-1, termed MGC-803-NRP(low), were generated from MGC-803 cells. Cell proliferation was analyzed by the Cell Counting Kit-8 and Bromodeoxyuridine incorporation assays, and migrating ability analyzed by migration assays. The xenograft model was used to assess the effects of NRP-1 depletion on tumorigenesis, growth, metastasis and therapeutic potentials. The role of NRP-1 as co-receptors in the signaling pathways stimulated by ligands was examined. The key molecules involved in cell proliferation, migration and related signaling pathways were detected by immunoblotting. RESULTS: Gastric cancer tissues expressed higher levels of NRP-1 compared to normal gastric mucosa. Its expression correlated with clinical staging, tumor differentiation and pathological types. NRP-1 depletion inhibited cell proliferation by inducing cell cycle arrest in the G1/S phase by upregulating p27, and downregulating cyclin E and cyclin-dependent kinase 2. NRP-1 depletion reduced the ability of cells to migrate by inhibiting the phosphorylation of focal adhesion kinase. NRP-1 depletion suppressed tumorigenesis, tumor growth and lung metastasis by inhibiting cell proliferation and tumor angiogenesis in situ. Therapeutic NRP-1 shRNA inhibited the growth of established BGC823 tumors. Depletion of NRP-1 inhibited the activation of VEGF/VEGFR2, EGF/EGFR and HGF/c-Met pathways stimulated by respective recombinant human VEGF-165, EGF and HGF proteins. CONCLUSIONS: The present results indicate that NRP-1 may be a potentially valuable biomarker and therapeutic target for gastric cancer.
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Proteína C-Reativa/genética , Proteína C-Reativa/metabolismo , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Neoplasias Gástricas/patologia , Animais , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Camundongos , Transplante de Neoplasias , Prognóstico , Transdução de Sinais , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Análise de SobrevidaRESUMO
Achalasia is a prototypic esophageal motility disorder with complications including aspiration-pneumonia, esophagitis, esophageal-tracheal fistula, spontaneous rupture of the esophagus, and squamous cell carcinoma. However, achalasia is rarely associated with esophageal stones and ulcer formation that lead to upper gastrointestinal bleeding. Here, we report the case of a 61-year-old woman who was admitted to our department after vomiting blood for six hours. Physical examination revealed that the patient had severe anemia and mild palpitation in the upper abdomen. CT revealed lower esophageal dilatation and esophageal wall thickening, and an emergency upper endoscopy showed that the esophagus was substantially expanded by a dark round stone, with multiple ulcers on the esophageal wall and a slit in the cardiac mucosa with a large clot attached. The patient's history included ingestion of 1 kg hawthorn three days prior. The acute upper gastrointestinal bleeding was caused by Mallory-Weiss syndrome associated with achalasia and an esophageal stone. For patients with achalasia, preventing excessive ingestion of tannins is crucial to avoid complications such as bleeding and rupture.
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In this paper, the problem of delay-dependent asymptotic stability analysis for neural networks with time-varying delays is considered. A new class of Lyapunov functional is proposed by considering the information of neuron activation functions adequately. By using the delay-partitioning method and the reciprocally convex technique, some less conservative stability criteria are obtained in terms of linear matrix inequalities (LMIs). Finally, two numerical examples are given to illustrate the effectiveness of the derived method.
Assuntos
Algoritmos , Retroalimentação , Modelos Teóricos , Redes Neurais de Computação , Dinâmica não Linear , Processamento de Sinais Assistido por Computador , Simulação por ComputadorRESUMO
BACKGROUND: Assessment of inflammatory activity in patients with ulcerative colitis (UC) is crucial to the prediction of relapse. Confocal laser endomicroscopy (CLE) is an accurate tool for assessing inflammatory activity in UC patients. This study aimed to evaluate whether CLE could be used to predict UC relapse reliably. METHODS: In total, forty-three patients with documented UC were analyzed in this study. Patients identified as having obvious active inflammation by conventional colonoscopy were excluded. The mucosa of each patient's sigmoid colon and rectum was assessed by CLE before targeted biopsies were taken. The patients were then followed up for at least 12 months to evaluate relapse according to the Simple Clinical Colitis Activity Index. The correlation between CLE classification and UC relapse was evaluated. RESULTS: Seventeen of 20 patients with histologically confirmed normal or chronic inflammation were diagnosed as having non-active inflammation by real-time CLE and 22 of 23 patients with histologically confirmed acute inflammation were diagnosed as having active inflammation by CLE. The sensitivity, specificity, and accuracy of CLE in real-time diagnosis of active inflammation were 95.7%, 85%, and 90.7%, respectively. The agreement between CLE and conventional histology was excellent (kappa value = 0.812). Two of 18 (11.1%) patients who were classified as having non-active inflammation by CLE relapsed, while 16 of 25 (64%) patients classified as having as active inflammation relapsed. The relapse rate of patients with active inflammation was significantly higher than of those with non-active inflammation (P < 0.001). CONCLUSIONS: CLE is comparable to conventional histology in predicting relapse in patients with UC.
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Colite Ulcerativa/patologia , Colo Sigmoide/patologia , Colonoscopia , Microscopia Confocal , Reto/patologia , Adulto , Idoso , Biópsia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Recidiva , Sensibilidade e Especificidade , Adulto JovemRESUMO
BACKGROUND: Acriflavine is one of the commonly used staining agents in confocal laser endomicroscopy (CLE), a newly developed technique allows for real time histological observation of gastrointestinal mucosa, but the concentration is not unified. This study aimed to evaluate the effects of acriflavine with different concentrations on the CLE image quality and to find a sound concentration in clinical practice. METHODS: Twenty four consecutive patients who underwent upper gastrointestinal CLE were enrolled into this study. The patients randomly accepted acriflavine in four different concentrations which were the conventional 0.05% and 3 lower ones respectively: 0.02%, 0.01% and 0.005% spraying onto the same focal antrum mucosa during CLE procedures. Differences of Image quality were demonstrated by an objective score system. RESULTS: THERE WAS NO SIGNIFICANT DIFFERENCE ABOUT IMAGE QUALITY AMONG ACRIFLAVINE CONCENTRATIONS: 0.05%, 0.02% and 0.01%, but 0.005% decreased image quality significantly (P=0.012). And 0.005% was also the only one which decreased general assessment significantly (P=0.01). For the 3 diagnostic value assessment indices, there was no significant difference about nonspecific and even staining, while 0.02% showed significant better polar staining (P=0.03). CONCLUSIONS: Acriflavine concentration 0.02% is the best one applied in CLE with the best nuclei staining ability and preserved image quality.
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OBJECTIVES: The assessment of inflammation activity in ulcerative colitis (UC) includes endoscopy and histology. Confocal laser endomicroscopy (CLE) combines real-time endoscopy and histology. This study was aimed at evaluating the application of CLE in the assessment of inflammation activity in UC. METHODS: In total, 73 consecutive patients with UC who visited Qilu Hospital for colonoscopy surveillance underwent CLE. Inflammation activity was first assessed by the colonoscopy Baron score, then by CLE with a 4-grade classification of crypt architecture, as well as by analysis of microvascular alterations and fluorescein leakage. Targeted biopsy samples were taken for histological analysis. Stored CLE images were subjected to post-CLE objective assessment. RESULTS: Both assessment of crypt architecture and fluorescein leakage with CLE showed good correlations with histological results (Spearman's rho, both P<0.001). CLE seemed to be more accurate than conventional white-light endoscopy for evaluating macroscopical normal mucosa. More than half of the patients with normal mucosa seen on conventional white-light endoscopy showed acute inflammation on histology, whereas no patients with normal mucosa or with chronic inflammation seen on CLE showed acute inflammation on histology. Assessment of microvascular alterations by CLE showed good correlation with histological findings (P<0.001). On post-CLE objective assessment, subjective architectural classifications were supported by the number of crypts per image (P<0.001) but not fluorescein leakage results by gray scale (P=0.194). CONCLUSIONS: CLE is reliable for real-time assessment of inflammation activity in UC. Crypt architecture, microvascular alterations, and fluorescein leakage are promising markers in CLE evaluation.
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Colite Ulcerativa/patologia , Colonoscopia/métodos , Inflamação/classificação , Microscopia Confocal , Adulto , Feminino , Humanos , Inflamação/patologia , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND AND AIM: Confocal laser endomicroscopy allows subsurface analysis of gastrointestinal mucosa during ongoing endoscopy. The present study assessed the feasibility of in vivo detecting superficial vascular architecture by confocal endomicroscopy in normal upper gastrointestinal mucosa and malignant lesions. METHODS: Early gastric cancer in eight patients, superficial esophageal carcinoma in six patients, and asymptomatic normal control in 10 patients were studied by confocal endomicroscopy. The characteristic of endomicroscopic microvascular architecture from normal and malignant mucosa was described and images were evaluated. RESULTS: Confocal endomicroscopy enabled clear visualization of the vascular networks of gastroesophageal mucosa. Honeycomb-like and coil-shaped regular microvascular architecture surrounding gastric pits were visible in the normal gastric body and antrum, respectively. Differentiated gastric cancerous mucosa showed hypervascularity and various caliber microvessels with irregular shapes. Undifferentiated gastric cancers disclosed a hypovascularity and irregular short branch vessels. Normal squamous epithelium had regular intraepithelial papillary capillary loops (IPCLs) directed toward the luminal surface. In superficial esophageal squamous carcinoma, dilated IPCLs were visible at the upper layer of the squamous mucosa. In esophageal adenocarcinoma, abnormal microvascular architecture showed tortuous and various calibers blood vessels. Of all the images, 41% were graded as good quality. The mean kappa value for interobserver agreement for the prediction of cancerous mucosa was 0.792. CONCLUSIONS: Confocal laser endomicroscopy system could yield very clear images of superficial microvascular network in the gastroesophageal mucosal layer both in malignant and normal mucosa. Endomicroscopic observation of vascular architecture may be of assistance in the identification of early gastroesophageal cancers.
Assuntos
Neoplasias Esofágicas/diagnóstico , Mucosa Gástrica/irrigação sanguínea , Microscopia Confocal , Neovascularização Patológica/diagnóstico , Neoplasias Gástricas/diagnóstico , Adenocarcinoma/diagnóstico , Carcinoma de Células Escamosas/diagnóstico , Estudos de Viabilidade , Humanos , MicrocirculaçãoRESUMO
OBJECTIVE: To evaluate the effectiveness and safety of mosapride on treatment of functional dyspepsia. METHODS: Randomized controlled clinical trial was conducted and patients suffered from functional dyspepsia were included. 5 mg mosapride was given three times daily for 4 weeks in the treatment group. 10 mg domperidone was given three times daily for 4 weeks as control. Changes on symptom score, gastric empty or new occurring events were included as outcomes. RESULTS: 231 patients suffered from functional dyspepsia were selected by inclusion and exclusion criteria from August 15 to Oct 22, 1999. Of these, 108 (46.8%) were males, versus 123 (53.2%) females and 118 (51.2%) in the treatment group and 113 (48.9%) as controls. 222 (96.1%) patients were followed up. Results showed that the total efficacy rates in early satiety and abdominal distension were 84.5% and 90.1% in mosapride after the 2 weeks of treatment. Mosapride seemed to be more effective in improving symptoms of belching and heartburn than that in controls (P < 0.05). In 4 weeks, the total efficacy in improving symptoms of abdominal distention and belching showed more effective in mosapride than that in controls (P < 0.05). Decrease of symptoms score was more in mosapride than that in controls (P < 0.05). Mosapride was less effective in controls in improving the gastric empty in terms of proportion (46.2% vs. 25.9%, P = 0.020) and range (46.2% vs. 24.0%, P = 0.003). Side effects would include diarrhea, constipation, headache, dizziness, insomnia, skin scare and the like. There was no significant difference between the two groups (9.6% in mosapride vs. 14.0% in controls). CONCLUSION: Mosapride was safe and effective in improving the symptoms and gastric empty of functional dyspepsia.