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Objective Establishment of orthotopic transplantation tumor model of human choriocarcinoma in nude mice. Methods Human choriocarcinoma cell line JAR was cultured and the single cell suspension was subcutaneously injected into five 8-week-old BALB / c nude mice to establish subcutaneous xenograft model. After subcutaneous tumor was formed in nude mice, the tumor tissue was taken under aseptic conditions and cut into 1 mm
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Objective To explore the role of TGF-β1 on epithelial mesenchymal transformation and invasion by promoting cancer stem cell marker CD133 expression in choriocarcinoma cells. Method Choriocarcinoma cells JEG-3 were cultured in vitro and incubated with TGF-β1 at different time and concentration, and the expression of CD133 protein and mRNA of EMT markers were detected by Western blotting and PCR respectively. The effect of TGF-β1 on invasive ability of JEG-3 cells were assessed with transwell method. Results TGF-β1 promoted the expression of cancer stem cell marker CD133, downregulated the epithelial marker E-cadherin, upregulated mesenchymal marker N-cadherin, and promoted invasion ability in choriocarcinoma cells. Conclusion TGF-β1 could promote stem cell property of cancer, EMT property, and invasive property of choriocarcinoma cells.
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PURPOSE: Periostin mediates critical steps in gastric cancer and is involved in various signaling pathways. However, the roles of periostin in promoting gastric cancer metastasis are not clear. The aim of this study was to investigate the relevance between periostin expression and gastric cancer progression and the role of stress-related hormones in the regulation of cancer development and progression. MATERIALS AND METHODS: Normal, cancerous and metastatic gastric tissues were collected from patients diagnosed with advanced gastric cancer. The in vivo expression of periostin was evaluated by in situ hybridization and immunofluorescent staining. Meanwhile, human gastric adenocarcinoma cell lines MKN-45 and BGC-803 were used to detect the in vitro expression of periostin by using quantitative real-time polymerase chain reaction (PCR) and western blotting. RESULTS: Periostin is expressed in the stroma of the primary gastric tumors and metastases, but not in normal gastric tissue. In addition, we observed that periostin is located mainly in pericryptal fibroblasts, but not in the tumor cells, and strongly correlated to the expression of α-smooth muscle actin (SMA). Furthermore, the distribution patterns of periostin were broader as the clinical staging of tumors progressed. We also identified a role of stress-related signaling in promoting cancer development and progression, and found that isoprenaline upregulated expression levels of periostin in gastric cancer cells. CONCLUSION: These findings suggest that the distribution pattern of periostin was broader as the clinical staging of the tumor progressed and found that isoprenaline upregulated expression levels of periostin in gastric cancer cells.
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Idoso , Humanos , Masculino , Adenocarcinoma/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Western Blotting , Moléculas de Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Fibroblastos/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Isoproterenol/farmacologia , Estadiamento de Neoplasias , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Estômago/metabolismo , Neoplasias Gástricas/metabolismo , Regulação para CimaRESUMO
Cardiovascular disease, with high morbidity and mortality, has been threatening the health of human beings. Therefore, expecting to find a more effective therapeutic method, a plenty of researchers devote themselves to the study of the cardiovascular disease all the time. Since discovered on the heart, M3 receptor of muscarinic acetylcholine receptor (mAchR, M receptor) became a new starting point of the research of the cardiovascular disease. With more and more investigation, many people found that M3 receptor could protect the heart from kinds of cardiovascular disease, which may make it a new hopeful therapeutic point. So, expecting to give support to the reference and encouragement for the study of disease related to M3 receptor in future, this review expounds M3 receptor on the heart from the main following aspects: the effect on the heart, the influence on the cardiovascular disease and the mechanism of M3 receptor involved.
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Humanos , Doenças Cardiovasculares , Coração , Fisiologia , Receptor Muscarínico M3 , FisiologiaRESUMO
Objective To investigated the effects of combined arsenic trioxide(ATO) and resveratrol(Res)on the viability of NB4 human leukemia cells. Methods NB4 human leukemia cell was used in this experiment.Cells were cultured in ATO (0,0.1875,0.3750,0.7500, 1.1250, 1.5000,2.2500,3.0000,5.0000 μmol/L) and Res (0, 1.5625,3.1250,6.2500, 12.5000, 18.7500,25.0000,37.5000,50.0000 μmol/L). Cell viabilities were measured by MTT in different treatment groups. Half inhibitory concentration(IC50) was calculated. The ratio of concentration of ATO and Res 1.5∶ 18,1.5∶ 25,1.5∶ 35 was added to cells, and the combination index(CI) was calculated. The level of ROS in control, ATO( 1.5000 μmol/L), Res(25.0000 μmol/L) and ATO(0.9000 μmol/L) + Res( 12.5000μmol/L) groups was measured by chemiluminescence assay. Results ①ATO( ≥0.7500 μmol/L) reduced the viability of NB4 cells in a concentration-dependent manner(P < 0.05 ), and IC50 was (1.78 ± 0.11 )μmol/L. ②)Res (≥18.7500 μ mol/L) dose-dependently decreased the viability of NB4 cells (P < 0.05 ), and IC50 was ( 18.71 ±0.18)μ mol/L. ③Combination of ATO and Res showed an antagonistic effect on NB4 cells viability. ④The ROS in Res group( 1670.55 ± 13.97) was significantly lower than that in control group(2345.88 ± 14.48,P < 0.05). The ROS in ATO group (3092.42 ± 94.84) was significantly higher than that in control group(P < 0.05). The ROS in ATO + Res group (1860.27 ± 15.99) was significantly lower than that in ATO group(P < 0.05). Conclusions NB4 cell survival rate can be decreased by ATO and Res. The combination of arsenic trioxide and Res presents an antagonistic effect on NB4 cell viability, in part by reducing intracellular ROS formation.
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To study the efficacy and tolerability of Daming capsule (DMC) in Chinese patients with hyperlipidemia, a randomized, multi-centre, open-label, parallel-group trial was conducted. Sixty enrolled patients with hyperlipidemia allocated to six medical centers were randomly divided into two groups of 30 individuals each. One group received DMC 2 g b.i.d. for 6 weeks, and the other received pravastatin 10 mg o.d. for 6 weeks. For efficacy assessment, serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) were measured before and after drug treatment. Serum TC and LDL-C levels in the DMC-treatment group were significantly decreased compared with those before treatment (p < 0.05), while TG and HDL-C levels did not change much. Tolerability was assessed by heart rate (HR), blood pressure (BP), body mass index (BMI), alanine aminotransferase (ALT) and creatinine (Cr), which were not changed in either the DMC or pravastatin groups at 3 and 6 weeks (p > 0.05). Besides, eight patients experienced diarrhea during DMC treatment and two experienced myalgia and epigastric discomfort during pravastatin treatment. Based on the above results, it was concluded that DMC may be a good candidate for the treatment of hyperlipidemia and further clinical trials are warranted.
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Anticolesterolemiantes/uso terapêutico , Medicamentos de Ervas Chinesas/uso terapêutico , Hipercolesterolemia/tratamento farmacológico , Pravastatina/uso terapêutico , Adulto , Alanina Transaminase/sangue , Pressão Sanguínea , Índice de Massa Corporal , Cápsulas , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Creatinina/sangue , Feminino , Frequência Cardíaca , Humanos , Masculino , Medicina Tradicional Chinesa , Triglicerídeos/sangueRESUMO
microRNAs are one kind of endogenous no-encoding RNA with about 22 nucleotides in length, and inhibited the translation of mRNAs by partially complementary binding to the 3' UTR of target mRNAs in the post-transcriptional level. Recent research shows that miRNAs function in the physiological and pathological processes of heart, especially involved in the occurrence and progress of arrhythmias. Abnormal miRNAs alters the protein expression of ion channels, causes the cardiac dysfunction, and triggers heart arrhythmias. The article summarized recent advances about roles of miRNA in arrhythmias and related cardiomyopathy, and discussed the therapeutic potential of miRNAs for heart diseases.
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Humanos , Arritmias Cardíacas , Genética , Metabolismo , Cardiomiopatias , Genética , Metabolismo , MicroRNAs , Genética , MetabolismoRESUMO
Human ether-a-go-go-related gene (HERG) encodes the rapid component of the cardiac delayed rectifier K+ current, which has an important effect on both proarrhythmia and antiarrhythmia. To investigate the effect of sophocarpine (SC) on HERG channel stably expressing in human embryonic kidney-293 (HEK293) cells, whole-cell patch-clamp technique was used to record HERG current and kinetic curves. As the result, it was found that SC inhibited HERG current in a concentration-dependent manner (10, 30, 100, and 300 micromol x L(-1)). At 0 mV, 10, 30, 100, and 300 micromol x L(-1) SC respectively inhibited IHERG by Istep ( 10.7 +/- 2.8)% , (11.3 +/- 5.5)% , (47.0 +/- 2.3)% and (53.7 +/- 2.5)% , and Itail (1.1 +/- 3.0)%, (17.1 +/- 3.3)%, (32.7 +/- 1.9)% (P < 0.05, n = 12) and (56.0 +/- 2.4)% (P < 0.05, n = 13). The time constants of inactivation, recovery from inactivation and onset of inactivation were accelerated. SC did not change other channel kinetics (activation and deactivation). It is concluded that SC inhibited the transfected HERG channels by influencing the inactivation state, which is the probable anti-arrhythmic mechanism.
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Humanos , Alcaloides , Farmacologia , Antiarrítmicos , Farmacologia , Linhagem Celular , Relação Dose-Resposta a Droga , Canais de Potássio Éter-A-Go-Go , Metabolismo , Fisiologia , Rim , Biologia Celular , Cinética , Potenciais da Membrana , Técnicas de Patch-Clamp , Plantas Medicinais , Química , Sophora , QuímicaRESUMO
This study is to explore whether the protective effect of resveratrol on ischemia-reperfusion injury is correlated with the structural and functional association between M3 receptor (M3 subtype of muscarinic acetylcholine receptor) and Cx43 (connexin 43 gap junction proteins). Immunoprecipitation, immunoblotting and immunofluorescence were applied to investigate whether resveratrol has an effect on structural and functional association between M3 and Cx43. The effect of resveratrol on electrocardiogram Lead II ex vivo in rats, SOD (superoxide dismutase) activity and MDA (malondialdehyde) content was also observed in order to evaluate the protective effect of resveratrol on ischemia-reperfusion injury. Resveratrol could restore the structural and functional association between M3 receptor and Cx43 gap junction proteins that was partially destroyed under ischemia-reperfusion injury. The phosphorylation and spatial distribution disturbances in Cx43 expression caused by ischemia-reperfusion injury were also restored. Also, the QRS duration, SOD activity and MDA content were restored. Resveratrol could restore the structural and functional association between M3 receptor and Cx43 gap junction proteins.
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Animais , Masculino , Ratos , Conexina 43 , Metabolismo , Eletrocardiografia , Coração , Técnicas In Vitro , Malondialdeído , Metabolismo , Traumatismo por Reperfusão Miocárdica , Metabolismo , Miocárdio , Metabolismo , Fosforilação , Substâncias Protetoras , Farmacologia , Distribuição Aleatória , Ratos Wistar , Receptor Muscarínico M3 , Metabolismo , Estilbenos , Farmacologia , Superóxido Dismutase , MetabolismoRESUMO
Because HERG potassium channel has important effects on both proarrhythmia and antiarrhythmia, we use immunofluorescence and Western blotting methods to detect the expression of HERG channel of HERG-HEK cells in different concentrations of matrine, oxymatrine and resveratrol. The findings showed that both matrine (1 micromol x L(-1) ) and oxymatrine ( 1micromol x L (-1) ) increased HERG channel expression ( n = 5, P < 0. 05 ) , while matrine (100 micromol x L(-1) ) decreased HERG channel expression ( n = 5, P < 0. 05), resveratrol didn't affect HERG channel expression. In conclusion, different concentrations of matrine and oxymatrine affect HERG channel expression, while there is no relationship between resveratrol and HERG channel expression. It provides a theoretical support for the safety and mechanism of anti-arrhythmic drugs.
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Humanos , Alcaloides , Farmacologia , Antiarrítmicos , Farmacologia , Western Blotting , Linhagem Celular , Relação Dose-Resposta a Droga , Canal de Potássio ERG1 , Canais de Potássio Éter-A-Go-Go , Genética , Metabolismo , Fisiologia , Imunofluorescência , Potenciais da Membrana , Técnicas de Patch-Clamp , Plantas Medicinais , Química , Quinolizinas , Farmacologia , Sophora , Química , Estilbenos , FarmacologiaRESUMO
MicroRNAs (miRNAs) are endogenous noncoding RNAs, about 22 nucleotides in length, that mediate post-transcriptional gene modulation by annealing to inexactly complementary sequences in the 3'-untranslated regions of target mRNAs. miRNA alterations are involved in the initiation and progression of human diseases. miRNA-expression profiling of human diseases has identified signatures associated with diagnosis, staging, progression, prognosis and response to treatment. Recent evidence has suggested miRNAs as viable therapeutic targets for a wide range of human diseases. Several approaches were performed, the experimental examination of these techniques and the resultant findings not only indicate feasibility of interfering miRNA action in a gene-specific fashion but also may provide a new research tool for studying function of miRNAs. The new approaches also have the potential of becoming alternative gene therapy strategies.
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Animais , Humanos , Desenho de Fármacos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Inativação Gênica , Terapia Genética , Métodos , Cardiopatias , Genética , Metabolismo , Terapêutica , Hipertensão , Genética , Metabolismo , Terapêutica , MicroRNAs , Genética , Metabolismo , Neoplasias , Genética , Metabolismo , Terapêutica , RNA Mensageiro , GenéticaRESUMO
<p><b>OBJECTIVE</b>To find the molecular mechanism of decreasing blood fat effect of Darning capsule on hyperlipemic rat, we study the expression of connexin43 in the myocardium before and after using the capsule.</p><p><b>METHOD</b>Forty Wistar rats were randomly divided into 5 group: control group, hyperlipemia model group, Daming capsule group of high dose, middle dose and low dose (200, 100, 50 mg kg(-1) d(-1)). Each group had 8 rats. Hyperlipemic rat model was made firstly, the blood was obtained via vena caudalis and the indexes of TC, TG, LDL, HDL and NEFA in the serum were measured. The myocardial total RNA was extracted by Trizol method. To compare the expression of connexin43 in the following groups: hyperlipemia, normal and drug, we used the technique of RT-PCR, immunostaining and microconfoul.</p><p><b>RESULT</b>The concentrations of TC, TG, LDL and NEFA in hyperlipemic serum were increased (P <0. 05), while that of HDL was decreased (P <0. 05). After treating with Daming capsule, the concentration of the preceding four indexes were decreased and the concentrations of HDL was increased up to nearly normal level. No significant difference was found in the ECG of the three groups. As compared with the normal group, the mRNA expressions of connexin43 in hyperlipemia group was weakened (P <0.05), while that of the drug group was enhanced(P <0.05). The same result in immunostaining was observed.</p><p><b>CONCLUSION</b>Hyperlipemic rat model was successfully established and Daming capsule has the effect of lowering blood lipid. Furthermore, the molecular mechanism of Darning capsule is related with the change of Cx43 closely.</p>
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Animais , Masculino , Ratos , Cápsulas , Colesterol , Sangue , Conexina 43 , Genética , Medicamentos de Ervas Chinesas , Farmacologia , Ácidos Graxos não Esterificados , Sangue , Hiperlipidemias , Sangue , Metabolismo , Hipolipemiantes , Farmacologia , Miocárdio , Metabolismo , Plantas Medicinais , Química , Isoformas de Proteínas , Genética , RNA Mensageiro , Distribuição Aleatória , Ratos Wistar , Triglicerídeos , SangueRESUMO
Viscum coloratum flavonoids (VCF) have been demonstrated to produce a variety of biological actions. An accumulating line of evidence supported the view that VCF may exert protective effects on the cardiovascular system. The aim of the study was to assess the antiarrhythmic activity as well as the electrophysiological properties of VCF. The antiarrhythmic effects of VCF were observed in a rat model of arrhythmia induced by aconitine. VCF significantly and dose-dependently increased the dosage of aconitine required to induce the arrhythmia indexes. Electrophysiological experiment revealed that VCF shortened APD through inhibition of ICa-L.
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Antiarrítmicos/farmacologia , Arritmias Cardíacas/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Flavonoides/farmacologia , Viscum/química , Aconitina/administração & dosagem , Aconitina/farmacologia , Potenciais de Ação/efeitos dos fármacos , Animais , Antiarrítmicos/química , Arritmias Cardíacas/induzido quimicamente , Canais de Cálcio/efeitos dos fármacos , Medicamentos de Ervas Chinesas/química , Flavonoides/química , Miócitos Cardíacos/efeitos dos fármacos , Distribuição Aleatória , Ratos , Ratos WistarRESUMO
<p><b>AIM</b>To optimize the method of investigating structural integration between proteins and study the integration between arrhythmia related proteins in molecular level.</p><p><b>METHODS</b>Immunostaining the normal ventricular myocytes was used to observe the distribution of connexin 43 and muscarinic acetylcholine receptor (mAChR). The five mAChR subtypes were precipitated using immunoprecipitation. Then, SDS-PAGE and Western blotting with the anti-connexin 43 antibody were performed to observe whether they were structurally integrated. Further, different concentrations of detergent were used to observe whether this relationship could be broken.</p><p><b>RESULTS</b>The five subtypes of mAChR existed in the cardiac myocyte of the rat, and all the five mAChR subtypes combined with connexin 43. In the normal rat ventricular myocyte membrane, connexin 43 and M3 receptor are co-located. When adding certain concentration of detergent to the membrane protein, the integration between M3 receptor and connexin 43 was broken, and the phosphorylated form of connexin 43 integrated with M3 receptor.</p><p><b>CONCLUSION</b>The results indicated that the structural integration between mAChR and phosphorylation of connexin 43 existed in rat ventricular myocardium, and this integration could be broken by certain concentration of detergent.</p>