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Homeobox-containing 1 (HMBOX1) has been described as a transcription factor involved in the occurrence of some tumors, but its roles in ovarian cancer have never been reported. Here we aimed to investigate the roles of HMBOX1 on high-grade serous ovarian carcinoma (HGSOC). In this present study, HMBOX1 expression was decreased in HGSOC tissues and ovarian cancer cell lines (HO8910 and A2780) compared with ovarian surface epithelial tissues or normal human ovarian surface epithelial cell line (HOSEpiC). The cell proliferation of HOSEpiC was weaker than ovarian cancer cell lines. By altering the expression of HMBOX1 in A2780 and HOSEpiC, we demonstrated that HMBOX1 inhibited the cell proliferation and promoted the cell apoptosis. Furthermore, our study revealed that HMBOX1 downregulated the expression of anti-apoptotic proteins (Bcl-2, Bcl-xL), raised the expression of pro-apoptotic-regulated proteins (Bad, Bax), apoptotic executionior (Caspase3), and P53. In conclusion, HMBOX1 played important roles in occurrence of HGSOC through regulation of proliferation and apoptosis, which implied that HMBOX1 might serve as a new therapeutic target for HGSOC.
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Cistadenocarcinoma Seroso/patología , Proteínas de Homeodominio/metabolismo , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/patología , Apoptosis/fisiología , Biomarcadores de Tumor/metabolismo , Estudios de Casos y Controles , Línea Celular Tumoral , Proliferación Celular/genética , Cistadenocarcinoma Seroso/genética , Cistadenocarcinoma Seroso/metabolismo , Regulación hacia Abajo , Femenino , Regulación Neoplásica de la Expresión Génica , Técnicas de Silenciamiento del Gen , Proteínas de Homeodominio/genética , Humanos , Neoplasias Ováricas/genéticaRESUMEN
BACKGROUND: Ovarian cancer is the most leading cause of death and the third most common gynecologic malignancy in women. Traditional chemotherapy has inevitable drawbacks of nonspecific tumor targeting, high toxicity, and poor therapeutic efficiency. In order to overcome such shortcomings, we prepared a novel nano-carrier drug-delivery system to enhance the anti-tumor efficiency. METHODS: In vitro characterizations of nano-carriers were determined by TEM, DLS. Cell viability was measured by MTT method. RT-PCR was performed to measure the expression of FARα in three ovarian cancer cell lines. The drug-release study and the uptaken study were measured in vitro. The pharmacokinetic and the drug distribution study were verified by HPLC methods in vivo. The enhanced anti-tumor efficiency of FA-NP was evaluated by the tumor inhibitory rate in vivo. RESULTS: Paclitaxel (PTX)-loaded nanoparticles (NPs) (PTX-PEG-PLA-NP and PTX-PEG-PLA-FA-NP) were prepared successfully, and the drug-release study showed that the cumulative release rates of NP groups were much less than free PTX group. The pharmacokinetic study showed that the elimination phase of two kinds of NP groups were much longer than that of PTX group. The drug distribution in different tissues showed that the peak-reach time was 2 h in the PTX group and 6 h in both NP groups. All of these results confirmed the excellent slow-release effects of both kinds of nano-carriers. More importantly, we confirmed that PTX-PEG-PLA-FA-NP had greater uptake by SK-OV-3 cells than PTX-PEG-PLA-NP and free PTX in vitro. A drug-distribution study of tumor-bearing mice demonstrated that the PTX concentration of tumor tissues in the PTX-PEG-PLA-FA-NP group was 3 times higher than the other two groups. PTX-PEG-PLA-FA-NP was uptaken much more by SK-OV-3 cells than PTX-PEG-PLA-NP and free PTX. Eventually, based on the slow-release effect and tumor-targeting characteristics of PTX-PEG-PLA-FA-NP, a cytotoxicity test indicated that PTX-PEG-PLA-FA-NP was much more toxic to SK-OV-3 cells than the controls. The tumor inhibitory rate in the PTX-PEG-PLA-FA-NP group of tumor-bearing mice was about 1.5 times higher than the controls. The tumor targeting and anti-tumor efficiency of PTX-PEG-PLA-FA-NP were confirmed both in vitro and in vivo. CONCLUSIONS: We developed an ovarian cancer targeting nano-carrier drug delivery system successfully, which showed perfect ovarian cancer targeting and anti-tumor effect, thus have the potential to be a new therapy strategy for ovarian cancer patients.
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Antineoplásicos Fitogénicos/administración & dosificación , Nanopartículas , Neoplasias Ováricas/patología , Paclitaxel/administración & dosificación , Nanomedicina Teranóstica , Animales , Antineoplásicos Fitogénicos/farmacocinética , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Liberación de Fármacos , Femenino , Humanos , Ratones , Terapia Molecular Dirigida , Nanopartículas/química , Neoplasias Ováricas/tratamiento farmacológico , Paclitaxel/farmacocinética , Polietilenglicoles/química , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
INTRODUCTION: The study aims to confirm the association of acute myocardial infarction (AMI) with serum angiotensin II (AngII), kallikrein1 (KLK1), and ACE/KLK1 polymorphisms. MATERIALS AND METHODS: Serum AngII/KLK1 levels and ACE and KLK1 genotypes were determined in 208 patients with AMI and 216 normal controls. Binary logistic regression was used for data analysis. RESULTS: The differences in serum AngII levels were statistically significant between the groups. After adjusting for potential confounding factors, high serum levels of AngII and KLK1 significantly increased the risk of AMI. The individuals with ACE DD and KLK1 GG genotypes significantly increased the risk of AMI compared with those harboring the ACE II and KLK1 AA genotypes (OR = 8.77, 95% CI = 1.74-44.16). CONCLUSIONS: (1) Increasing the serum levels of AngII increased the risk of AMI. (2) The risk of AMI increased significantly when the serum levels of AngII and KLK1 simultaneously increased. (3) Individuals with the combined genotypes of ACE DD and KLK1 GG showed significantly increased risk of AMI compared with those with the combined genotypes of ACE II and KLK1 AA.
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Angiotensina II/sangre , Estenosis Coronaria/complicaciones , Infarto del Miocardio/etiología , Infarto del Miocardio/genética , Peptidil-Dipeptidasa A/genética , Polimorfismo de Nucleótido Simple/genética , Calicreínas de Tejido/sangre , Calicreínas de Tejido/genética , Estudios de Casos y Controles , Distribución de Chi-Cuadrado , Estenosis Coronaria/sangre , Femenino , Frecuencia de los Genes , Humanos , Mutación INDEL/genética , Modelos Logísticos , Masculino , Persona de Mediana Edad , Infarto del Miocardio/sangre , Reacción en Cadena de la PolimerasaRESUMEN
OBJECTIVE: To investigate the effects of mifepristone, a progesterone receptor (PR) antagonist, through the proliferation of human cholangiocarcinoma cell line FRH-0201 in vitro and the possible mechanisms involved. METHODS: A two-step addition of poly-HRP anti-mouse immunoglobulin G detection system was used to detect the expression of PR in FRH-0201 cells. After treatments with various concentrations of mifepristone (10, 20, 40, 80, 160, and 320 µmol/L) at various time intervals (24, 48, and 72 hours), the rate of cell inhibition, the rate of cell apoptosis, and the expression of bax/bcl-2/Fas were analyzed with tetrazolium blue (MTT) assay, flow cytometry, reverse transcription polymerase chain reaction and Western blotting. The effect of mifepristone and mifepristone combined with interferon (IFN)-γ-inducing apoptosis on the cells was observed. RESULTS: Mifepristone remarkably inhibited the proliferation of FRH-0201 cells, which was revealed by MTT assay in a dose- and time-dependent manner. The inhibitory rate gradually increased following the increase of the dosage of mifepristone from a low dosage (10 µmol/L) to a high dosage (320 µmol/L) at different time intervals. Flow cytometry analysis showed mifepristone increased the rate of the FRH-0201 cell-line apoptosis. Notably, the rate of apoptosis increased markedly when the cells were pretreated with IFN-γ and then treated with mifepristone. In addition, mifepristone obviously upregulated bax and Fas expression and downregulated bcl-2 expression. CONCLUSION: Mifepristone effectively inhibited the growth of PR-positive human cholangiocarcinoma cell line FRH-0201 in vitro through multiple mechanisms. Mifepristone combined with IFN-γ might therefore induce the apoptosis of the cell line, which is possibly a beneficial clinical scheme for patients suffering from cholangiocarcinoma.
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BACKGROUND: Quercetin has been shown to induce apoptosis in a number of cancer cell lines, but a quercetin-loaded nanoliposomal formulation with enhanced antitumor activity in C6 glioma cells and its effect on cancer cell death has not been well studied. The aim of this study was to examine if quercetin-loaded liposomes (QUE-NL) has enhanced cytotoxic effects and if such effects involve type III programmed cell death in C6 glioma cells. METHODS: C6 glioma cells were treated with QUE-NL and assayed for cell survival, apoptosis, and necrosis. Levels of reactive oxygen species production and loss of mitochondrial membrane potential (ΔΨm) were also determined by flow cytometry assay to assess the effects of QUE-NL. ATP levels and lactate dehydrogenase activity were measured, and Western blotting was used to assay cytochrome C release and caspase expression. RESULTS: QUE-NL induced type III (necrotic) programmed cell death in C6 glioma cells in a dose-dependent and time-dependent manner. High concentrations of QUE-NL induced cell necrosis, which is distinct from apoptosis and autophagy, whereas liposomes administered alone induced neither significant apoptosis nor necrosis in C6 glioma cells. QUE-NL-induced ΔΨm loss and cytochrome C release had no effect on caspase activation, but decreased ATP levels and increased lactate dehydrogenase activity indicated that QUE-NL stimulated necrotic cell death. CONCLUSION: C6 glioma cells treated with QUE-NL showed a cellular pattern associated with necrosis without apoptosis and was independent of caspase activity. Nonapoptotic cell death induced by high concentrations of QUE-NL for controlling caspase-independent type III programmed cell death may provide the basis for novel therapeutic approaches to overcome avoidance of apoptosis by malignant cells.
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Glioma/tratamiento farmacológico , Liposomas/farmacología , Nanopartículas/química , Quercetina/farmacología , Adenosina Trifosfato/metabolismo , Animales , Antioxidantes/química , Antioxidantes/farmacología , Apoptosis/efectos de los fármacos , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Línea Celular Tumoral , Forma de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Citocromos c/metabolismo , Glioma/patología , L-Lactato Deshidrogenasa/metabolismo , Liposomas/química , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Necrosis , Quercetina/química , Ratas , Especies Reactivas de Oxígeno/metabolismoRESUMEN
This study aimed to investigate the association of the aldehyde dehydrogenase 2 (ALDH2) Glu504Lys polymorphism, which exists in 30-50% of East Asians, and risk of acute coronary syndrome (ACS). We enrolled 1092 unrelated Han Chinese, including 546 with ACS and 546 age- and sex-matched controls. Subjects with ALDH2 mutant genotypes showed significantly higher ACS than did controls (46.7% versus 31.9%, P < 0.001). Logistic regression analysis revealed the ALDH2 mutant independently associated with ACS (odds ratio [OR] 1.95, 95% confidence interval [CI]: 1.31-2.92, P = 0.001), but the association was weaker on adjusting for alcohol consumption (OR 1.82, 95% CI: 1.23-2.70, P = 0.003). Similar results were found in a subgroup analysis of patients with primary ST-segment elevation myocardial infarction (STEMI). The ALDH2 mutant was significantly associated with level of high-sensitivity C-reactive protein (hs-CRP) in patients with ACS (P = 0.002) and in controls (P = 0.009) and number of circulating endothelial progenitor cells (EPCs) (P = 0.032); furthermore, inclusion of hs-CRP level and EPCs number as independent variables in regression analysis reduced the importance of ALDH2 polymorphism in ACS or primary STEMI. However, ALDH2 polymorphism was not associated with number of coronary arteries with significant stenosis, Gensini score or flow-mediated dilation of the brachial artery. Our results suggest that ALDH2 mutation is a genetic risk marker for ACS, which is explained in part by alcohol consumption, inflammation and number of circulating EPCs.
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Síndrome Coronario Agudo/enzimología , Síndrome Coronario Agudo/genética , Aldehído Deshidrogenasa/genética , Sustitución de Aminoácidos/genética , Predisposición Genética a la Enfermedad , Polimorfismo de Nucleótido Simple/genética , Síndrome Coronario Agudo/diagnóstico por imagen , Síndrome Coronario Agudo/fisiopatología , Aldehído Deshidrogenasa Mitocondrial , Arteria Braquial/fisiopatología , Proteína C-Reactiva/metabolismo , Estudios de Casos y Controles , Recuento de Células , Movimiento Celular , Demografía , Células Endoteliales/citología , Células Endoteliales/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Infarto del Miocardio/diagnóstico por imagen , Infarto del Miocardio/enzimología , Infarto del Miocardio/genética , Infarto del Miocardio/fisiopatología , Células Madre/citología , Células Madre/metabolismo , Ultrasonografía , Vasodilatación/fisiologíaRESUMEN
BACKGROUND: It has been demonstrated that biliopancreatic diversion (BPD) and ileal transposition (IT) effectively induce weight loss and long-term control of type 2 diabetes in morbidly obese individuals. It is unknown whether the control of diabetes is better after IT or after BPD. The objective of this study was to investigate the effects of IT and BPD on the control of diabetes in an animal model. METHODS: We performed IT and BPD on 10- to 12-week-old Goto-Kakizaki rats with a spontaneous nonobese model of type 2 diabetes, and we performed a series of detection. The rats were observed for 24 weeks after surgery. RESULTS: Animals who underwent IT and BPD demonstrated improved glucose tolerance, insulin sensitivity and the secretion of glucagon-like peptide-1 compared with the sham-operated animals. Furthermore, IT resulted in a shorter duration of surgery and better postoperative recovery than BPD. CONCLUSION: This study provides strong evidence for the crucial role of the hindgut in the resolution of diabetes after duodenum-jejunum bypass or IT. We confirmed that IT was associated with better postoperative recovery than BPD and had a similar control of diabetes as BPD in nonobese animals with type 2 diabetes.
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Diabetes Mellitus Experimental/cirugía , Diabetes Mellitus Tipo 2/cirugía , Íleon/cirugía , Anastomosis Quirúrgica , Animales , Desviación Biliopancreática , Glucemia/metabolismo , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Péptido 1 Similar al Glucagón/metabolismo , Resistencia a la Insulina/fisiología , Masculino , RatasRESUMEN
OBJECTIVE: To investigate the low density lipoprotein receptor (LDLR) gene and apolipoprotein (Apo) B gene mutation in a Chinese family with familial hypercholesterolemia (FH) and give the kindreds clinical check-ups. METHODS: After physical examination, the kindreds underwent ECG and ultrasound checks. Blood samples were tested for lipid profiles. The promoter and all eighteen exons of LDLR gene were investigated by using PCR and agarose gel electrophoresis in combination with DNA sequence analysis. The results were compared with the normal sequences in GenBank and FH database (www.ucl.ac.uk/fh) to find mutations. In addition, the apolipoprotein B100 gene for known mutations (R3500Q, R3531C, R3501W and R3480W) that cause familial defective ApoB100 (FDB) was also tested using the same method. RESULTS: A novel homozygous G > A mutation at the 1581 bp of exon 10 was detected in the proband and his siblings. It caused a substitution of amino acid Glu to Gly at codon 496. A novel heterozygous G > A mutation at the 1581 bp of exon 10 was detected in his parents. No mutations of R3500Q, R3531C, R3501W and R3480W of ApoB100 were observed. ECGs were normal. Atherosclerosis were found in all family members by ultrasound checks. CONCLUSIONS: The homozygous G > A mutation at the 1581 bp of exon 10 was first determined in our country. The change of amino acid Glu to Gly is responsible for FH of the family. The type of the gene mutation was not found in the FH database (www. ucl.ac.uk/fh). It's a new type of LDLR mutation.
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Hiperlipoproteinemia Tipo II/genética , Mutación , Receptores de LDL/genética , Adolescente , Adulto , Apolipoproteína B-100/sangre , Apolipoproteína B-100/genética , Exones , Femenino , Humanos , Hiperlipoproteinemia Tipo II/sangre , Masculino , Persona de Mediana Edad , Linaje , Fenotipo , Receptores de LDL/sangre , Adulto JovenRESUMEN
The reduced expression of angiotensin-converting enzyme (ACE) 2 in the kidneys of animal models and patients with diabetes suggests ACE2 involvement in diabetic nephrology. To explore the renoprotective effects of ACE2 overexpression, ACE inhibition (ACEI) or both on diabetic nephropathy and the potential mechanisms involved, 50 Wistar rats were randomly divided into a normal group that received an injection of sodium citrate buffer and a diabetic model group that received an injection of 60 mg/kg streptozotocin. Eight wks after streptozotocin injection, the diabetic rats were divided into no treatment group, adenoviral (Ad)-ACE2 group, Ad-green flurescent protein (GFP) group, ACEI group receiving benazepril and Ad-ACE2 + ACEI group. Four wks after treatment, physical, biochemical, and renal functional and morphological parameters were measured. An experiment in cultured glomerular mesangial cells was performed to examine the effects of ACE2 on cellular proliferation, oxidative stress and collagen IV synthesis. In comparison with the Ad-GFP group, the Ad-ACE2 group exhibited reduced systolic blood pressure, urinary albumin excretion, creatinine clearance, glomeruli sclerosis index and renal malondialdehyde level; downregulated transforming growth factor (TGF)-ß1, vascular endothelial growth factor (VEGF) and collagen IV protein expression; and increased renal superoxide dismutase activity. Ad-ACE2 and ACEI had similar effects, whereas combined use of Ad-ACE2 and ACEI offered no additional benefits. ACE2 transfection attenuated angiotensin (Ang) II-induced glomerular mesangial cell proliferation, oxidative stress and collagen IV protein synthesis. In conclusion, ACE2 exerts a renoprotective effect similar to that of ACEI treatment. Decreased renal Ang II, increased renal Ang-(1-7) levels, and inhibited oxidative stress were the possible mechanisms involved.
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Inhibidores de la Enzima Convertidora de Angiotensina/uso terapéutico , Nefropatías Diabéticas/tratamiento farmacológico , Nefropatías Diabéticas/metabolismo , Nefropatías Diabéticas/patología , Glomérulos Renales/metabolismo , Peptidil-Dipeptidasa A/genética , Peptidil-Dipeptidasa A/metabolismo , Angiotensina II/metabolismo , Angiotensina II/farmacología , Enzima Convertidora de Angiotensina 2 , Animales , Glucemia/efectos de los fármacos , Presión Sanguínea/efectos de los fármacos , Línea Celular , Proliferación Celular/efectos de los fármacos , Diabetes Mellitus Experimental , Modelos Animales de Enfermedad , Expresión Génica , Técnicas de Transferencia de Gen , Vectores Genéticos , Glomérulos Renales/citología , Glomérulos Renales/patología , Masculino , Estrés Oxidativo , Ratas , Ratas WistarRESUMEN
AIMS: Adventitial fibroblasts (AFs) and inflammation play an important role in neointimal formation and vascular remodeling. The present study was aimed to investigate the therapeutic effects and underlying mechanisms of transcriptional regulator Gax gene transfection in aortic remodeling induced by adventitial inflammation. METHODS AND RESULTS: Fifty rabbits fed a chow diet were randomly divided into a normal control group (n=10) and experimental group (n=40). All rabbits in the experimental group underwent collar placement around the abdominal aorta and intra-collar injection of lipopolysaccharide (LPS) to induce adventitial inflammation and they were further divided into model control group, saline-treated group, green fluorescence protein (Ad-GFP)-treated group and Gax gene (Ad-Gax)-treated group, respectively. Four weeks after treatment, the model control group, saline-treated group and Ad-GFP-treated group showed thickened neointima and adventitia, reduced lumen size and increased eccentricity and remodeling index of the abdominal aorta in comparison with the normal control group, whereas Ad-Gax-treated group exhibited attenuated neointimal formation and vascular remodeling (P<0.01-0.05) .The vascular expression levels of interleukin (IL)-1ß, IL-6, IL-8, monocyte chemoattractant protein (MCP)-1, matrix metalloproteinase (MMP)-1, MMP-2, MMP-9, vascular cell adhesion molecule (VCAM)-1 and intercellular adhesion molecule (ICAM)-1, Smads, mitogen-activated protein kinases (MAPKs), integrins and nuclear factor kappa B (NF-kB) were significantly higher in the model control group, saline-treated group and Ad-GFP-treated group than those in the normal control group (P<0.01-0.05). In contrast, the local expression levels of these cytokines were substantially reduced by Ad-Gax gene transfer (P<0.01-0.05). Similarly, the serum levels of inflammatory cytokines including C-reactive protein (CRP), transforming growth factor (TGF)-ß1, IL-1, IL-6, IL-8, tumor necrosis factor (TNF)-α, MCP-1, VCAM-1 and ICAM-1 were significantly higher in the model control group, saline-treated group and Ad-GFP-treated group than those of the Ad-Gax-treated group (P<0.01-0.05). In vitro studies showed that Gax overexpression diminished inflammatory cytokine expression in LPS-stimulated arterial fibroblasts. CONCLUSIONS: Adventitial inflammation induces vascular remodeling via the interactions of multiple inflammatory cytokines and local Gax gene transfer in vivo can significantly inhibit these interactions and thereby attenuate local inflammation and vascular remodeling.
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Vasos Sanguíneos/patología , Proteínas de Homeodominio/genética , Adenoviridae/genética , Animales , Aorta/metabolismo , Tejido Conectivo/patología , Citocinas/metabolismo , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Humanos , Inflamación , Masculino , Conejos , Transducción de Señal , Ultrasonografía/métodosRESUMEN
BACKGROUND: Metabolic and inflammatory pathways crosstalk at many levels. In this study, we aimed to investigate the expression of six-transmembrane protein of prostate 2 (STAMP2) in macrophages and tried to search for the association between the decreased STAMP2 expression, if any, and carotid atherosclerosis as well as cardiac adaptations. MATERIALS AND METHODS: A total of 97 unrelated Chinese subjects were recruited including 48 subjects with metabolic syndrome (MetS) and 49 controls. Clinical and biochemical characteristics were collected from subjects, with quantification of STAMP2 in monocyte/macrophages. All subjects underwent ultrasonography. RESULTS: STAMP2 expression in macrophages was significantly decreased in MetS as compared with the control group (10.25 +/- 9.20 vs. 15.20 +/- 9.18, P = 0.009), especially in women patients. Partial correlation analysis showed that STAMP2 expression in macrophages correlated with BMI (r = -0.375, P = 0.045), age (r = 0.414, P = 0.026) and HDL (r = 0.377, P = 0.044) after controlling for systolic blood pressure (SBP). Furthermore, STAMP2 expression was correlated with PI (r = -0.454, P = 0.013), LVEF (r = -0.503, P = 0.005), LA-ESR (r = -0.424, P = 0.022), LA-S (r = 0.469, P = 0.010) and mitral E/A ratio (r = 0.492, P = 0.005) after controlling for SBP. Still, in multivariable analysis, STAMP2 expression was independently associated with IMT(mean), PI and mitral E/A ratio. CONCLUSIONS: In MetS patients, especially women patients, STAMP2 expression was down-regulated in peripheral blood mononuclear cell, which was correlated with carotid atherosclerosis and cardiac adaptation.
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Arterias Carótidas/patología , Enfermedades de las Arterias Carótidas/metabolismo , Proteínas de la Membrana/metabolismo , Síndrome Metabólico/metabolismo , Monocitos/metabolismo , Oxidorreductasas/metabolismo , Factores de Edad , Pueblo Asiatico , Aterosclerosis/metabolismo , Índice de Masa Corporal , Arterias Carótidas/diagnóstico por imagen , Femenino , Humanos , Lipoproteínas HDL/análisis , Macrófagos/metabolismo , Masculino , Persona de Mediana Edad , Análisis Multivariante , Factores de Riesgo , Factores Sexuales , Ultrasonografía , Función Ventricular Izquierda/fisiologíaRESUMEN
OBJECTIVES: High-risk human papillomaviruses (HPVs) are the major causative agents of cervical cancer, and the E6 and E7 genes encode the major HPV oncoproteins. The E7 protein of high-risk HPV types disturbs cell cycle control and down-regulates components of the antigen presentation pathway, suggesting a role for E7 in tumor immune evasion. We previously reported that HPV-16 E7 expression and down-regulation of HLA class I was highly correlated in cervical lesions. This study was aimed to determine whether HPV-16 E7 oncoprotein could down-regulate surface HLA class I antigen in HPV-16 E7-transfected cells, and whether it had correlation with the expression of the transporter associated with antigen processing (TAP). METHODS: The HPV-16 E7 open reading frame was transfected into HaCaT cells. After G418 selection, resistant colonies were individually picked and expanded into clonal cell lines. Using the fluoresence-activated cell sorting analysis, the levels of cell surface HLA class I antigen and intracellular TAP-1 and TAP-2 expressions were detected. RESULTS: Compared with the empty vector control, a statistical significant decrease of approximately 50% in cell surface HLA class I expression was observed in HPV-16 E7 expressing HaCaT cells (P < 0.001). Moreover, the expression of HPV-16 E7 in HaCaT cells resulted in decreased expression of TAP-1 that was essential for HLA class I expression at the cell surface, a statistical significant decrease of approximately 40% compared with that with the empty vector control (P < 0.001). CONCLUSIONS: Our finding demonstrates that HPV-16 E7 down-regulates surface HLA class I antigen, which in part correlates with the decrease of TAP-1.
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Transportadoras de Casetes de Unión a ATP/metabolismo , Regulación Neoplásica de la Expresión Génica , Antígenos de Histocompatibilidad Clase I/metabolismo , Proteínas E7 de Papillomavirus/metabolismo , Transportador de Casetes de Unión a ATP, Subfamilia B, Miembro 2 , Miembro 3 de la Subfamilia B de Transportadores de Casetes de Unión a ATP , Línea Celular , Regulación hacia Abajo , Humanos , Sistemas de Lectura Abierta , Proteínas E7 de Papillomavirus/genética , TransfecciónRESUMEN
This study was designed to identify the differential expression of the canonical transient receptor potential (TRPC) channels in the left ventricle of spontaneously hypertensive rats (SHR). Echocardiography studies were performed to compare the left ventricular function in SHR vs. Wistar-Kyoto rats (WKY), and the mRNA level of the TRPC channels was determined by quantitative real-time RT-PCR (qRT-PCR). Western blots were performed to examine whether the mRNA expression corresponded with the protein expression. Compared with the WKY, the mRNA expression of TRPC4 and TRPC5 was significantly increased in the 10-week-old SHR (P = 0.032 for TRPC4 and P = 0.043 for TRPC5), so did the TRPC4/5 protein content. The midwall fractional shortening (mFS) of SHR was lower than WKY (P = 0.016). Furthermore, increased expression of TRPC4/5 was correlated with both increased blood pressure and decreased mFS. These findings suggest that TRPC4 and 5 seem to be the main subtypes expressed in the heart of the SHR at the beginning period of hypertension. Theses channels may participate in the development of left ventricular systolic dysfunction.
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Ventrículos Cardíacos/metabolismo , Canales de Potencial de Receptor Transitorio/genética , Canales de Potencial de Receptor Transitorio/metabolismo , Animales , Presión Sanguínea/fisiología , Electrocardiografía , Electroforesis en Gel de Agar , Regulación de la Expresión Génica , Frecuencia Cardíaca/fisiología , Ventrículos Cardíacos/fisiopatología , Masculino , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: Angiotensin converting enzyme 2 (ACE2) efficiently hydrolyses the potent vasoconstrictor angiotensin II to vasodilative angiotensin (1-7). We hypothesized that ACE2 overexpression may inhibit inflammation response in atherosclerotic plaque by degrading Ang II into Ang-(1-7). METHODS: Atherosclerosis (AS) plaques were induced in the abdominal aorta of 38 rabbits by endothelial injury and atherogenic diet for 3 months. Rabbits were then underwent injection of a recombinant adenovirus (2.5 x 10(9) pfu/ml) carrying a murine ACE2 gene (Ad-ACE2) through a catheter into the abdominal aortic segments rich in plaques (n = 19) or injection of a control vector Ad-EGFP (n = 19). One month later, all rabbits were sacrificed and plaques from aortic segments were analyzed. RESULTS: ACE2 expression in aortic tissues of the Ad-ACE2 group were confirmed by immunohistochemistry. Macrophage infiltration (13.6% +/- 4.2% vs. 23.6% +/- 6.9%, P < 0.01) and MCP-1 expression (13.2% +/- 0.4% vs. 25.0% +/- 7.4%, P < 0.01) were significantly reduced in Ad-ACE2 group compared to Ad-EGFP group. CONCLUSIONS: Overexpression of ACE2 inhibited atherosclerotic plaque inflammation response in hypercholesterolemic rabbits.
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Aterosclerosis/genética , Peptidil-Dipeptidasa A/genética , Transfección , Enzima Convertidora de Angiotensina 2 , Animales , Aterosclerosis/metabolismo , Células Cultivadas , Dieta Aterogénica , Vectores Genéticos , ConejosRESUMEN
This study was carried out to test the hypothesis that Tongxinluo (TXL) as a Chinese herbal medicine enhances stability of vulnerable plaque dose dependently via lipid-lowering and anti-inflammation effects, similar to a high-dose simvastatin therapy. After abdominal aortic balloon injury, 75 rabbits were fed a 1% cholesterol diet for 10 wk and were then divided into five groups for 8-wk treatment: control group, low-dose TXL group, moderate-dose TXL group, high-dose TXL group, and high-dose simvastatin group. At the end of week 16, an adenovirus containing p53 was injected into the abdominal aortic plaques. Two weeks later, plaque rupture was induced by pharmacological triggering. The incidence of plaque rupture in all treatment groups (14.3%, 7.1%, 7.7%, and 7.1%) was significantly lower than that in control group (73.3%; P>0.01). TXL dose-dependently lowered serum lipid levels and inhibited systemic inflammation. Corrected acoustic intensity and fibrous cap thickness of the aortic plaques were significantly increased, whereas plaque area, plaque burden, vulnerable index, and expression of oxidized low-density lipoprotein (ox-LDL) receptor 1, matrix metalloproteinase 1 (MMP-1), MMP-3, tissue inhibitor of MMP 1, and NF-kappaB in plaques were markedly reduced in all treatment groups when compared with the control group. Similar to high-dose simvastatin group, high-dose TXL group exhibited a low serum level of low-density lipoprotein cholesterol and ox-LDL, a low expression level of systemic and local inflammatory factors and a low plaque vulnerability index, with no differences in the incidence of plaque rupture among all treatment groups. TXL dose-dependently enhances the stability of vulnerable plaques and prevents plaques from rupture. Simvastatin and TXL offer similar protection in terms of lipid-lowering, anti-inflammation, and antioxidation effects.
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Antiinflamatorios/farmacología , Antioxidantes/farmacología , Enfermedades de la Aorta/tratamiento farmacológico , Rotura de la Aorta/prevención & control , Aterosclerosis/tratamiento farmacológico , Medicamentos Herbarios Chinos/farmacología , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Simvastatina/farmacología , Animales , Enfermedades de la Aorta/etiología , Enfermedades de la Aorta/genética , Enfermedades de la Aorta/metabolismo , Enfermedades de la Aorta/patología , Rotura de la Aorta/etiología , Rotura de la Aorta/genética , Rotura de la Aorta/metabolismo , Rotura de la Aorta/patología , Aterosclerosis/etiología , Aterosclerosis/genética , Aterosclerosis/metabolismo , Aterosclerosis/patología , Biomarcadores/metabolismo , Cateterismo/efectos adversos , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Relación Dosis-Respuesta a Droga , Regulación de la Expresión Génica/efectos de los fármacos , Técnicas de Transferencia de Gen , Hipercolesterolemia/complicaciones , Hipercolesterolemia/tratamiento farmacológico , Hipercolesterolemia/patología , Inmunohistoquímica , Mediadores de Inflamación/metabolismo , Lípidos/sangre , Masculino , ARN Mensajero/metabolismo , Conejos , Factores de Tiempo , Proteína p53 Supresora de Tumor/genética , Ultrasonografía Doppler Dúplex , Ultrasonografía Intervencional , Venenos de VíborasRESUMEN
Because both Toll-like receptor-1 (TLR1) and TLR2 are expressed in atherosclerotic plaques, we hypothesized that TLR1 and TLR2 may play different roles in the formation of vulnerable plaques and that combinatorial knockdown of TLR1 and TLR2 genes may enhance the effects of isolated knockdown of the TLR1 or TLR2 gene on plaque stabilization. Lentiviruses carrying small interfering RNAs of TLR1 or TLR2 were constructed, which knocked down mRNA and protein expression of TLR1 or TLR2 significantly in vitro. One hundred and forty apolipoprotein E-deficient (apoE(-/-)) mice were randomly allocated to control, mock, TLR1 interference (TLR1i), TLR2 interference (TLR2i), and TLR1+2 interference (TLR1+2i) subgroups and a constrictive collar was placed around the carotid artery of these mice to induce plaque formation. TLR1i and TLR2i viral suspension was transfected into the carotid plaques separately in the TLR1i and TLR2i subgroups or together in the TLR1+2i subgroup. Four weeks after lentivirus transfection, expression of both TLR1 and TLR2 in the carotid plaques was remarkably attenuated. Plaques of the TLR1i subgroup showed lower macrophage content and interleukin (IL)-6 expression and a thicker fibrous cap compared with the control or mock subgroups. Plaques of the TLR2i subgroup showed a higher content of collagen and lower content of lipid and macrophages, a thicker fibrous cap, lower vulnerability index, and lower mRNA expression of IL-6 and monocyte chemoattractant protein-1 than the TLR1i subgroup. In the TLR1+2i subgroup, the macrophage and smooth muscle cell content, and the vulnerability index, were ameliorated as compared with those in the TLR2i subgroup. Lentivirus-mediated RNA interference can be used to efficiently knock down TLR1 and TLR2 genes in carotid plaques of apoE(-/-) mice. Although isolated knockdown of TLR1 or TLR2 is effective in attenuating plaque vulnerability, combinatorial interference with TLR1 and TLR2 exhibits enhanced improvement of plaque stability, and thus provides a useful approach to the stabilization of vulnerable plaques.
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Apolipoproteínas E/deficiencia , Aterosclerosis/patología , Silenciador del Gen , Receptor Toll-Like 1/genética , Receptor Toll-Like 2/genética , Animales , Apolipoproteínas E/sangre , Aterosclerosis/sangre , Aterosclerosis/complicaciones , Western Blotting , Peso Corporal , Línea Celular , Citocinas/genética , Citocinas/metabolismo , Regulación de la Expresión Génica , Humanos , Inflamación/complicaciones , Inflamación/patología , Lentivirus/genética , Ratones , Ratones Noqueados , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Receptor Toll-Like 1/metabolismo , Receptor Toll-Like 2/metabolismo , TransfecciónRESUMEN
BACKGROUND: WWOX and FHIT are two candidate tumor suppressor genes located in active fragile sites, the damage of which has been associated with the development of breast cancer. The association of the expression of these genes and the development of breast cancer has not been fully explored. We evaluated mRNA and protein expression of WWOX and FHIT in breast tissue with normal histological appearances, atypical ductal hyperplasia, ductal carcinoma in situ, and invasive cancer to see if a progressive decline in expression was present. METHODS: Reverse transcription-polymerase chain reaction and Western blotting were used to evaluate the specimens for mRNA and protein expression, including 28 specimens with normal tissue, 28 specimens with atypical ductal hyperplasia, 33 specimens with ductal carcinoma in situ, and 51 specimens with invasive ductal carcinoma. RESULTS: Compared with in situ and invasive cancer specimens, both normal and atypical hyperplasia specimens had greater rates of detectable mRNA (WWOX rate ratio = 2.95, 95% CI 1.24 - 7.08; FHIT rate ratio = 4.58, 95% CI 1.82 - 11.81) and Western blotting detectable protein (WWOX rate ratio = 4.12, 95% CI 1.63 - 10.73; FHIT rate ratio = 3.76, 95% CI 1.44 - 10.06). For both proteins, differences between normal and atypical hyperplasia specimens and between in situ and invasive carcinoma specimens were explainable by chance (P > 0.05 for each analysis). Within each histological category, differences among fractions of specimens showed that FHIT and WWOX mRNA and protein expression were explainable by chance (P > 0.05 for each analysis). CONCLUSION: Expression of FHIT and WWOX decreases along with breast tissue progress from a normal histological appearance to atypical ductal hyperplasia, in situ cancer, and the final invasive cancer.
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Ácido Anhídrido Hidrolasas/genética , Neoplasias de la Mama/genética , Mama/patología , Sitios Frágiles del Cromosoma , Genes Supresores de Tumor , Proteínas de Neoplasias/genética , Oxidorreductasas/genética , Proteínas Supresoras de Tumor/genética , Ácido Anhídrido Hidrolasas/análisis , Femenino , Humanos , Hiperplasia , Proteínas de Neoplasias/análisis , Oxidorreductasas/análisis , Proteínas Supresoras de Tumor/análisis , Oxidorreductasa que Contiene Dominios WWRESUMEN
BACKGROUND: The cause of Hirschsprung's disease (HD) remains unclear, but currently there are two theories: the mutation of the RET gene and the change of enteric microenvironment. This study was undertaken to elucidate the cause of HD by assessing the expression of laminin (LN), laminin gene, and the RET gene in the aganglionic segment, transitional zone and normal segment of the colon in patients with HD. METHODS: Specimens of the aganglionic segment, transitional zone, and normal segment of the colon from 27 cases of HD were stained immunohistologically by a PV 9000 polymer detection system. Photos were taken by the RS image system, and the staining area of each image was calculated by a JD 801 image analysis system. The qualitative expressions of the laminin gene and RET gene of these three segments in the 27 cases were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the difference of the expressions was shown by the alpha 9900 image analysis system. The quantitative expressions of the laminin gene and RET gene in the three segments were detected by real-time quantitative PCR, and the difference of the expression was shown by SDS software. RESULTS: The laminin and laminin gene were expressed in all the three segments. The expression was higher in the aganglionic segment than in the dilated segment, and the expression decreased stepwisely from the aganglionic segment to the normal segment, while the expression of the RET gene was opposite, showing an increased segmenting from the aganglionic segment to the normal segment. The correlation between the expressions of the two genes was negatively correlated. CONCLUSIONS: The highly increased expression of LN in the aganglionic segment may cause early differentiation, early maturation and premature ecesis of enteric nervous cells. The change of the microenvironment of colon wall may be the cause of HD. The negative correlation between the expression of the two genes may be closely related to the occurrence of HD.