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Purpose: The inflammatory response was associated with the prognosis of head and neck squamous cell carcinoma (HNSCC). This study aimed to perform a novel prognostic signature based on inflammation-related genes (IRGs) for a better understanding of the prognosis of HNSCC. Patients and Methods: IRGs were obtained from The Cancer Genome Atlas (TCGA) database and the Gene Expression Omnibus (GEO) database. Functional enrichment analysis was performed to explore potential pathways. Univariate and multivariate Cox regression as well as the Least Absolute Shrinkage and Selection Operator (LASSO) were utilized to construct an IRGs-based prognostic model on TCGA database and the GEO database was utilized for outcome validation. The nomogram model was constructed based on independent prognostic factors after univariate and multivariate Cox regression. The immune cell infiltration level was analyzed via the Tumor Immune Estimation Resource (TIMER) database. Results: In this study, we confirmed that 60% IRGs were abnormally expressed in HNSCC samples, and these were associated with important oncobiology. Then, a prognostic signature comprising 7 hub genes was generated based on TCGA database. The results were validated in 97 patients from GSE41613. A nomogram comprising risk score, age, M stage and N stage was generated to improve the accuracy of prognosis evaluation. The immune cell infiltration analysis suggested that 5 hub genes (ADGRE1, OLR1, TIMP1, GPR132 and CCR7) were negatively correlated with tumor purity and positively correlated with the infiltration of immune cells. Conclusion: Our study established a novel signature consisting of 7 hub genes for the prognostic prediction in patients with HNSCC.
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OBJECTIVES: The study aimed to investigate the effects of resveratrol on colorectal cancer HCT116 cells, including cell viability, apoptosis, and migration, and the partial mechanisms focused on hedgehog/gli-1 signaling pathways. MATERIALS AND METHODS: We chose the appropriate time and concentration of recombinant human Sonic hedgehog (Shh) stimulation by cell viability. The proportion of cell apoptosis was detected by flow cytometry; HCT116 cell migration was measured by scratch test; the expression of Ptch, Smo, and Gli-1 was measured by Western blot analysis. RESULTS: Shh signaling increased HCT116 cell viability and migration, inhibited cell apoptosis, and upregulated the expression of Ptch, Smo, and Gli-1. Resveratrol obviously inhibited HCT116 cell viability and migration, promoted cell apoptosis, and suppressed the protein of Ptch, Smo, and Gli-1. Furthermore, the effects of resveratrol and Shh on human colorectal cancer HCT116 cells were in a dose- and time-dependent manner. CONCLUSION: The inhibitory effect of resveratrol on HCT116 cells may be mediated by hedgehog/gli-1 signaling pathways.
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As the primary physiological barrier, intestinal epithelial cells regulate the transportation of oral therapeutic agents including nanomedicines which significantly improves the bioavailability of many drugs. However, currently there seems in the lack of comprehensive understanding on nanoparticle transport in intestinal epithelial cells as well as the mechanisms related. So, in an attempt to illustrate the profile of nanoparticle transport in intestinal epithelial cells, Caco-2 cells and polymer nanoparticles (PNs) were used as the models to explore the whole transport process including endocytosis, intracellular trafficking, exocytosis and transcytosis. Via various techniques, the transport pathways of PNs in Caco-2 cells and their mechanisms were clarified. Firstly, the transport was characterized by its non-specificity. The co-mediation of clathrin, lipid raft/caveolae and macropinocytosis as well as the co-involvement of different proteins like actins, protein tyrosine kinase (PTK) and cyclooxygenase (COX) were found in the endocytosis of PNs. The endocytosed PNs could transport to apical early endosome (AEE) and then from AEE to lysososmes via AEE/late endosome (LE)/lysosome pathway, as well as to recycling endosome compartment (REC) or endoplasmic reticulum (ER) through AEE/REC and AEE/ER pathways, respectively. Both ER/Golgi and Golgi/REC/plasma membrane (PM) pathways were involved in the exocytosis of PNs. The transcytosis of PNs across the cell monolayer was very low with a ratio less than 0.5%, due to complicated reasons. Secondly, the transport was evidenced by its partial energy-dependency. Beside the energy-dependent transport mediated by some proteins, quantitative study demonstrated the obvious internalization as well as surface binding of PNs at both 37 °C and 4 °C, but significantly higher at 37 °C. Interestingly, the consistency between surface binding and internalization at each temperature was found, suggesting that cell binding was the precondition and key step for the following endocytosis. The involvement of both energy dependent and independent mechanism was also observed in the exocytosis and transcytosis process of PNs. Finally, there were opposite mechanisms found between the exocytosis and endocytosis of PNs, including the regulation role of lipid raft/caveolae, COX and Golgi complex, which also contributed to the fact of "easy entry and hard across" for PNs. Overall, this study depicts a clear picture of nanoparticle transport in Caco-2 epithelial cells characterized by non-specificity, partial energy-dependency and low transcytosis.
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Nanopartículas/química , Nanopartículas/metabolismo , Polímeros/química , Células CACO-2 , Endocitosis , Exocitosis , Humanos , TranscitosisRESUMEN
Epithelial cell membranes as the typical biological barrier constitute the prime obstacle for the transport of therapeutic agents including nanomedicines. The previous studies on the interaction between nanomedicines and cells are mostly emphasized on cellular uptake and intracellular trafficking, but seldom on epithelial cells, although more and more oral nanomedicines are available now. In an attempt to clarify the transport pathways of nanomedicines in epithelial cells, the different molecular mechanisms among endocytosis, exocytosis and transcytosis processes were carefully studied and compared here using a kind of polymer nanoparticles (PNs) and MDCK epithelial cells as models. As the result, their similarity and difference were demonstrated. The similarities among all the three processes included the mediation of lipid rafts, the involvement of some protein kinases such as protein tyrosine kinase (PTK), protein kinase C (PKC) and phosphatidylinositol 3-kinase (PI3K), and the existence of multiple pathways. However, the difference among these processes was very significant, including different pathways, and especially the disparate effects of lipid rafts and protein kinases for different processes. The endocytosis involved both lipid raft and clathrin mechanisms but no macropinocytosis, via the invagination of membrane but no pore formation, the exocytosis contained ER/Golgi and Golgi/PM pathways, and transcytosis included AEE/CE/BSE and Golgi/BSE pathways. The roles of lipid rafts on endocytosis were positive but that on exocytosis and transcytosis was negative. The impacts of PTK and PKC on endocytosis were positive, while the influences of PTK, PKC and P13K on AEE/CE/BSE, as well as PTK and P13K on Golgi/BSE transcytosis pathways were negative. Moreover, the discrepancy between inward and outward transport of PNs elucidated an interesting fact that the endocytosis was rather easy and outward transport including exocytosis and transcytosis was rather difficult. Finally, it was indicated by comparison with previous reports that the molecular mechanisms between PNs and macromolecules such as proteins were also dissimilar.
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Células Epiteliales/metabolismo , Nanopartículas/química , Polímeros/metabolismo , Animales , Transporte Biológico/efectos de los fármacos , Cumarinas/farmacología , Perros , Endocitosis/efectos de los fármacos , Células Epiteliales/citología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/enzimología , Exocitosis/efectos de los fármacos , Citometría de Flujo , Espacio Intracelular/efectos de los fármacos , Espacio Intracelular/metabolismo , L-Lactato Deshidrogenasa/metabolismo , Luz , Células de Riñón Canino Madin Darby , Microscopía Confocal , Modelos Biológicos , Nanopartículas/ultraestructura , Orgánulos/efectos de los fármacos , Orgánulos/metabolismo , Tamaño de la Partícula , Proteínas Quinasas/metabolismo , Dispersión de Radiación , Tiazoles/farmacologíaRESUMEN
Sorafenib is slightly absorbed in the gastrointestinal tract due to its poor solubility in water. To improve its absorption, a novel nanoparticulate formulation-nanomatrix was used in the study. The nanomatrix was a system prepared from a porous material Sylysia(®) 350 and a pH sensitive polymer Eudragit(®). The formulations were optimized by orthogonal design (L(9)(3(4))) and their bioavailability were evaluated in rat, comparing to pH-sensitive Eudragit nanoparticles and suspension of sorafenib. In the formulations, the ratio of sorafenib to Eudragit(®) S100 was found to be more important determinant of the sorafenib bioavailability than the ratio of sorafenib to Sylysia(®) 350. As for the bioavailability, the AUC(0-36 h) of sorafenib nanomatrix was 13-33 times to that of sorafenib suspension, but only 16.8% to 40.8% that of Eudragit(®) S100 nanoparticles. This may be resulted from the different drug dispersion degree, release character and bioadhension activity. However, because all the materials used in the nanomatrix formulation are commonly adjuvant, safe, easy to get and cheap, above all, the nanomatrix formulation can solve the stability and scaling up problems in the nanoparticles, it had potential to develop into a product in the future.
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Antineoplásicos/administración & dosificación , Bencenosulfonatos/administración & dosificación , Nanopartículas , Piridinas/administración & dosificación , Adhesividad , Administración Oral , Animales , Antineoplásicos/farmacocinética , Área Bajo la Curva , Bencenosulfonatos/farmacocinética , Disponibilidad Biológica , Concentración de Iones de Hidrógeno , Masculino , Nanoestructuras , Niacinamida/análogos & derivados , Compuestos de Fenilurea , Ácidos Polimetacrílicos/química , Porosidad , Piridinas/farmacocinética , Ratas , Ratas Sprague-Dawley , Dióxido de Silicio/química , Solubilidad , Sorafenib , SuspensionesRESUMEN
A novel solid particle system with a nanomatrix structure and without surfactant for the oral delivery of insoluble drugs was prepared. This used a combination of pH-sensitive polymethylacrylate and nano-porous silica, in order to improve the drug absorption using only pharmaceutical excipients and a relative simple process. The in vitro drug dissolution and in vivo oral bioavailability of this formulation, using fenofibrate as the model drug, were compared with other reference formulations such as a suspension, micronized formulation or self microemulsion drug delivery system (SMEDDS). The supersaturation stabilizing effect of different polymers was evaluated and the physicochemical characterization of the optimal formulation was conducted by SEM, TEM, surface area analysis, DSC, and XRD. The optimized formulation prepared with polymethylacrylate (Eudragit®L100-55) and silica (Sylysia®350) markedly improved the drug dissolution compared with other reference preparations and displayed a comparative oral bioavailability to the SMEDDS. Fenofibrate existed in a molecular or amorphous state in the nanomatrix, and this state was maintained for up to 1year, without obvious changes in drug release and absorption. In conclusion, the nanomatrix formulation described here is a promising system to enhance the oral bioavailability of water-insoluble drugs.
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Sistemas de Liberación de Medicamentos , Fenofibrato/química , Hipolipemiantes/química , Nanoporos , Polimetil Metacrilato/química , Dióxido de Silicio/química , Absorción , Animales , Disponibilidad Biológica , Coloides/química , Evaluación Preclínica de Medicamentos , Estabilidad de Medicamentos , Emulsiones/química , Excipientes/química , Fenofibrato/administración & dosificación , Fenofibrato/farmacocinética , Concentración de Iones de Hidrógeno , Hipolipemiantes/administración & dosificación , Hipolipemiantes/farmacología , Absorción Intestinal/fisiología , Masculino , Tamaño de la Partícula , Polímeros/química , Ácidos Polimetacrílicos/química , Polimetil Metacrilato/metabolismo , Ratas , Ratas Sprague-Dawley , Solubilidad , Propiedades de Superficie , Tensoactivos/metabolismoRESUMEN
OBJECTIVE: Our aim was to investigate the effect of soy isoflavone (SI) on liver fibrosis in a thioacetamide (TAA)-induced rat model. MATERIALS AND METHODS: Twenty-eight rats were assigned to four groups: sham group, fibrosis group, low-dose treatment group (LDg) and high-dose treatment group (HDg). SI (90 or 270 mg/kg) was administered daily during the model development by TAA. Standard liver tests, platelet derived growth factor-BB (PDGF-BB) and tissue inhibitor of metalloproteinase-1 (TIMP-1) were measured. The expression of collagen, α-smooth muscle actin (α-SMA) and transforming growth factor-ß1 (TGF-ß1) in liver tissue was determined. Electron microscopy was used to perform ultrastructural analysis of the livers. RESULTS: Hepatic fibrosis was induced by 8 weeks of TAA administration. However, following the administration of SI, collagen staining significantly declined as compared with the fibrosis group (p < 0.01). Less collagen fibers around the hepatic stellate cells (HSCs) were observed in HDg as compared to the fibrosis group and LDg. There was no significant difference in standard liver tests between the fibrosis group and the two treatment groups. The levels of PDGF-BB and TIMP-1 in the two SI-treated groups were significantly lower than in the fibrosis group (p < 0.01). The expression of α-SMA and TGF-ß1 in HDg was less than that in the fibrosis group and LDg (p < 0.01). CONCLUSION: Administration of a high dose of SI resulted in an obvious inhibitory effect on liver fibrosis induced by TAA in rats. One hypothesis is that the effect may be related to the inhibition of HSC activation and proliferation.
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Glycine max , Isoflavonas/uso terapéutico , Cirrosis Hepática Experimental/prevención & control , Actinas/biosíntesis , Animales , Becaplermina , Colágeno/biosíntesis , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Isoflavonas/administración & dosificación , Hígado/citología , Hígado/metabolismo , Hígado/fisiopatología , Cirrosis Hepática Experimental/inducido químicamente , Cirrosis Hepática Experimental/metabolismo , Cirrosis Hepática Experimental/patología , Pruebas de Función Hepática , Masculino , Factor de Crecimiento Derivado de Plaquetas/análisis , Proteínas Proto-Oncogénicas c-sis , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tioacetamida/administración & dosificación , Tioacetamida/toxicidad , Inhibidor Tisular de Metaloproteinasa-1/análisis , Factor de Crecimiento Transformador beta1/biosíntesis , Resultado del TratamientoRESUMEN
AIM: To evaluate the clinical presentation, treatment and survival of patients with primary malignant tumor of small bowel (PMTSB). METHODS: Clinicopathologic data about 141 surgically treated PMTSB patients (91 males and 50 females) at the median age of 53.5 years (range 23-79 years) were retrospectively analyzed. RESULTS: The most common initial clinical features of the patients were intermittent abdominal discomfort or vague abdominal pain (67.4%), abdominal mass (31.2%), bowel obstruction (24.1%), hemotochezia (21.3%), jaundice (16.3%), fever (14.2%), coexistence of bowel perforation and peritonitis (5.7%), coexistence of gastrointestinal bleeding and shock (5.0%), and intraabdominal bleeding (1.4%). Ileum was the most common site of tumor (44.7%), followed by jejunum (30.5%) and duodenum (24.8%). PMTSB had a nonspecific clinical presentation. Segmental bowel resection (n = 81) was the most common surgical procedure, followed by right hemi-colectomy (n = 15), pancreaticoduodenectomy (n = 10), and others (n = 19). Twenty-seven adenocarcinoma patients and 13 malignant lymphoma patients received adjuvant chemotherapy with 5-fluorouracil and cyclophosphamide, adriamycin, vincristine and prednisone, respectively. Information about 120 patients was obtained during the follow-up. The median survival time of PMTSB patients was 20.3 mo. The 1-, 3- and 5-year survival rate was 75.0% (90/120), 40.0% (48/120) and 20.8% (25/120), respectively. Adenocarcinoma was found in 73.7% (42/57), 21.1% (12/57) and 15.8% (9/57) of the patients, respectively. Gastrointestinal stromal tumor was observed in 80.0% (20/25), 72.0% (18/25) and 36.0% (9/25) of the patients, respectively. Carcinoid was detected in 100.0% (15/15), 80.0% (12/15) and 46.7% (7/15) of the patients, respectively. Malignant lymphoma was demonstrated in 69.2% (9/13), 30.8% (4/13) and 0% (0/13) of the patients, respectively. CONCLUSION: En bloc resection is the principal therapy for most PMTSB and chemotherapy is the important treatment modality for malignant lymphoma and other malignant tumors of small bowel which cannot be radically removed.