RESUMEN
BACKGROUND: Post-cholecystectomy diarrhea (PCD) frequently occurs in patients following gallbladder removal. PCD is part of the post-cholecystectomy (PC) syndrome, and is difficult to treat. After cholecystectomy, bile enters the duodenum directly, independent of the timing of meals. The interaction between the bile acids and the intestinal microbes is changed. Therefore, the occurrence of PCD may be related to the change in microbiota. However, little is known about the relationship between the gut microbiota and PCD. AIM: To better understand the role of the gut microbiota in PCD patients. METHODS: Fecal DNA was isolated. The diversity and profiles of the gut microbiota were analyzed by performing high-throughput 16S rRNA gene sequencing. The gut microbiota were characterized in a healthy control (HC) group and a PC group. Subsequently, the PC group was further divided into a PCD group and a post-cholecystectomy non-diarrhea group (PCND) according to the patients' clinical symptoms. The composition, diversity and richness of microbial communities were determined and compared. RESULTS: In the PC and HC groups, 720 operational taxonomic units (OTUs) were identified. The PC group had fewer OTUs than the HC group. ß-diversity was decreased in the PC group. This indicated decreased microbial diversity in the PC group. Fifteen taxa with differential abundance between the HC and PC groups were identified. In the PCD group compared to the PCND group, significant decreases in microbial diversity, Firmicutes/Bacteroidetes ratio, and richness of probiotic microbiota (Bifidobacterium and Lactococcus), and an increase in detrimental microbiota (Prevotella and Sutterella) were observed. Moreover, a negative correlation was found between Prevotella and Bifidobacterium. Using a Kyoto Encyclopedia of Genes and Genomes functional analysis, it was found that the abundances of gut microbiota involved in lipid metabolism pathways were markedly lower in the PCD group compared to the PCND group. CONCLUSION: This study demonstrated that gut dysbiosis may play a critical role in PCD, which provides new insights into therapeutic options for PCD patients.
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Microbioma Gastrointestinal , Colecistectomía , Diarrea/etiología , Disbiosis , Humanos , ARN Ribosómico 16S/genéticaRESUMEN
Increasing levels of plasma urotensin II (UII) are positively associated with atherosclerosis. In this study we investigated the role of macrophage-secreted UII in atherosclerosis progression, and evaluated the therapeutic value of urantide, a potent competitive UII receptor antagonist, in atherosclerosis treatment. Macrophage-specific human UII-transgenic rabbits and their nontransgenic littermates were fed a high cholesterol diet for 16 weeks to induce atherosclerosis. Immunohistochemical staining of the cellular components (macrophages and smooth muscle cells) of aortic atherosclerotic lesions revealed a significant increase (52%) in the macrophage-positive area in only male transgenic rabbits compared with that in the nontransgenic littermates. However, both male and female transgenic rabbits showed a significant decrease (45% in males and 31% in females) in the smooth muscle cell-positive area compared with that of their control littermates. The effects of macrophage-secreted UII on the plaque cellular components were independent of plasma lipid level. Meanwhile the wild-type rabbits were continuously subcutaneously infused with urantide (5.4 µg· kg-1· h-1) using osmotic mini-pumps. Infusion of urantide exerted effects opposite to those caused by UII, as it significantly decreased the macrophage-positive area in male wild-type rabbits compared with that of control rabbits. In cultured human umbilical vein endothelial cells, treatment with UII dose-dependently increased the expression of the adhesion molecules VCAM-1 and ICAM-1, and this effect was partially reversed by urantide. The current study provides direct evidence that macrophage-secreted UII plays a key role in atherogenesis. Targeting UII with urantide may promote plaque stability by decreasing macrophage-derived foam cell formation, which is an indicator of unstable plaque.
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Aterosclerosis/tratamiento farmacológico , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Macrófagos/efectos de los fármacos , Fragmentos de Péptidos/farmacología , Placa Aterosclerótica/tratamiento farmacológico , Urotensinas/farmacología , Animales , Aterosclerosis/metabolismo , Aterosclerosis/patología , Células Cultivadas , Dieta Alta en Grasa/efectos adversos , Relación Dosis-Respuesta a Droga , Femenino , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Infusiones Subcutáneas , Macrófagos/metabolismo , Masculino , Fragmentos de Péptidos/administración & dosificación , Fragmentos de Péptidos/sangre , Placa Aterosclerótica/metabolismo , Placa Aterosclerótica/patología , Conejos , Urotensinas/administración & dosificación , Urotensinas/sangreRESUMEN
The results of several new clinical trials that compared the effectiveness of entecavir (ETV) treatment with that of adefovir (ADV) treatment in patients with chronic hepatitis B (CHB) were published in recent years. However, the numbers of patients included in these clinical trials were too small to draw a clear conclusion as to whether ETV is more effective than ADV. Therefore, a new meta-analysis was needed to compare ETV with ADV for the treatment of CHB. A search of the Cochrane Central Register of Controlled Trials (CCTR), MEDLINE, the Science Citation Index, Embase, the China National Knowledge Infrastructure (CNKI), and the Wanfang Database for relevant studies published between 1966 and 2010 was performed. Trials comparing the use of ETV and ADV for the treatment of CHB were assessed. Of the 2,358 studies screened, 13 randomized controlled clinical trials comprising 1,230 patients (ETV therapy, 621; ADV therapy, 609) were analyzed. The serum hepatitis B virus (HBV) DNA clearance rate obtained in patients treated with ETV was significantly higher than that in patients treated with ADV at the 24th and 48th weeks of treatment (24 weeks: 59.6% vs. 31.8%, relative risk [RR], 1.82, 95% CI: 1.49-2.23; 48 weeks: 78.3% vs. 50.4%, RR, 1.61, 95% CI: 1.32-1.96). The serum HBeAg clearance rate, the HBeAg seroconversion rate, and the ALT normalization rate obtained for patients treated with ETV were also higher than the corresponding values for patients treated with ADV at the 48th week of treatment. The safety profiles were similar between patients treated with ETV and those treated with ADV. The evidence reviewed in this meta-analysis suggests that patients with hepatitis B have a greater likelihood of achieving a viral response and a biomedical response when treated with ETV than when treated with ADV.
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Adenina/análogos & derivados , Antivirales/uso terapéutico , Guanina/análogos & derivados , Hepatitis B Crónica/tratamiento farmacológico , Organofosfonatos/uso terapéutico , Adenina/uso terapéutico , Guanina/uso terapéutico , Humanos , Ensayos Clínicos Controlados Aleatorios como AsuntoRESUMEN
Recent accumulating evidence has indicated that ZNF804A (zinc finger protein 804A) may be one of the most robustly implicated genes in schizophrenia. In this report, we examined ZNF804A single nucleotide polymorphisms (SNPs) encompassing exon 4 by performing an association study that used a Han Chinese sample comprised of 492 schizophrenia patients and 516 healthy control subjects. A meta-analysis based on previous studies was also performed. For markers rs4667000 and rs1366842, significant differences in allele frequencies were found between cases and controls (Mantel-Haenszel corrected P=0.014 and P=0.025, respectively). Analysis of haplotype rs61739290-rs1366842 showed significant association with schizophrenia (global P=0.0018). Moreover, several other two-, three-, and four-SNP tests of haplotype association were also significant. A meta-analysis comprised of studies that utilized sample sets of either European and/or Han Chinese origin revealed statistically significant associations for two SNPs (rs1366842, P=0.002; and rs3731834, P=0.03) and schizophrenia. In addition, we observed a significant association between marker rsl344706 and schizophrenia (P<1.0×10(-5)) in combined populations. When we separately analyzed the studies by population, consistent and significant differences were found between cases and controls both in the European samples (P<1.0×10(-4)) and in the Chinese samples (P=0.03). In summary, we have added new evidence supporting the association between ZNF804A and schizophrenia in our Han Chinese sample. Further functional exploration of ZNF804A will greatly help us to elucidate the pathogenesis of schizophrenia and find promising new approaches for the treatment of this disorder.
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Predisposición Genética a la Enfermedad , Factores de Transcripción de Tipo Kruppel/genética , Polimorfismo de Nucleótido Simple/genética , Esquizofrenia/genética , Adolescente , Adulto , Pueblo Asiatico/etnología , Pueblo Asiatico/genética , Femenino , Frecuencia de los Genes , Estudios de Asociación Genética , Genotipo , Humanos , Masculino , Población Blanca/genética , Adulto JovenRESUMEN
The results of several new clinical trials that compared the effectiveness of entecavir (ETV) treatment with that of adefovir (ADV) treatment in patients with chronic hepatitis B (CHB) were published in recent years. However, the numbers of patients included in these clinical trials were too small to draw a clear conclusion as to whether ETV is more effective than ADV. Therefore, a new meta-analysis was needed to compare ETV with ADV for the treatment of CHB. A search of the Cochrane Central Register of Controlled Trials (CCTR), MEDLINE, the Science Citation Index, Embase, the China National Knowledge Infrastructure (CNKI), and the Wanfang Database for relevant studies published between 1966 and 2010 was performed. Trials comparing the use of ETV and ADV for the treatment of CHB were assessed. Of the 2,358 studies screened, 13 randomized controlled clinical trials comprising 1,230 patients (ETV therapy, 621; ADV therapy, 609) were analyzed. The serum hepatitis B virus (HBV) DNA clearance rate obtained in patients treated with ETV was significantly higher than that in patients treated with ADV at the 24th and 48th weeks of treatment (24 weeks: 59.6% vs. 31.8%, relative risk [RR], 1.82, 95% CI: 1.49-2.23; 48 weeks: 78.3% vs. 50.4%, RR, 1.61, 95% CI: 1.32-1.96). The serum HBeAg clearance rate, the HBeAg seroconversion rate, and the ALT normalization rate obtained for patients treated with ETV were also higher than the corresponding values for patients treated with ADV at the 48th week of treatment. The safety profiles were similar between patients treated with ETV and those treated with ADV. The evidence reviewed in this meta-analysis suggests that patients with hepatitis B have a greater likelihood of achieving a viral response and a biomedical response when treated with ETV than when treated with ADV.
Asunto(s)
Humanos , Adenina/análogos & derivados , Antivirales/uso terapéutico , Guanina/análogos & derivados , Hepatitis B Crónica/tratamiento farmacológico , Organofosfonatos/uso terapéutico , Adenina/uso terapéutico , Guanina/uso terapéutico , Ensayos Clínicos Controlados Aleatorios como AsuntoRESUMEN
OBJECTIVE: To assess whether the TaqIB polymorphism of cholesteryl ester transfer protein (CETP) is associated with coronary artery disease (CAD) in Chinese population, we performed a meta-analysis in this paper. METHODS: We searched PubMed, Embase, the Science Citation Index (SCI), the China Biological Medicine database (CBM), the China National Knowledge Infrastructure (CNKI), and the Wanfang database for relevant articles. Data were extracted, and pooled odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. RESULTS: The literature search yielded 448 studies, in which 10 case-control studies including 1694 cases and 1456 controls matched the selection criteria. The combined B1 and B2 allele frequencies were 0.587 and 0.413, respectively. The pooled OR was 1.10 (95% CI, 0.89-1.34) for comparing the B1B1 or B1B2 carriers with B2B2 carriers, and was 1.27 (95% CI, 1.09-1.49) in the B1B1 carriers versus B2B2 or B1B2 carriers. CONCLUSIONS: In the present study, the TaqIB polymorphism of CETP was found to be associated with CAD in the Chinese population.
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Proteínas de Transferencia de Ésteres de Colesterol/genética , Enfermedad de la Arteria Coronaria/epidemiología , Enfermedad de la Arteria Coronaria/genética , Predisposición Genética a la Enfermedad/epidemiología , Predisposición Genética a la Enfermedad/genética , Heterocigoto , Polimorfismo de Nucleótido Simple/genética , Adulto , Anciano , Anciano de 80 o más Años , China/epidemiología , Femenino , Estudios de Asociación Genética , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Medición de Riesgo , Factores de Riesgo , Adulto JovenRESUMEN
Matrine is the major active component of the traditional Chinese medicine Sophora flavescens, but the molecular mechanisms of matrine on tumor invasion inhibition remain unclear. The aim of this study is to elucidate the effects of matrine on invasion ability of human hepatocellular carcinoma (HCC) cells, matrix metalloproteinase-9 (MMP-9), and nuclear factor (NF)-kappa B expression. The expression activity of MMP-9 was measured by reverse transcription polymerase chain reaction, Western blot, and gelatin zymography analysis. The expression of NF-kappa B was measured by the Western blot analysis. Matrine significantly inhibited MMP-9 expression of SMMC-7721 cells. NF-kappa B inhibitor PTDC induced a marked reduction in MMP-9 expression, and it suggested that NF-kappa B could play an important role in MMP-9 expression. Furthermore, matrine significantly suppressed NF-kappa B expression and the invasion of SMMC-7721 cells. Our results showed that matrine inhibited MMP-9 expression and the invasion of human HCC cells. The inhibitory effects are partly associated with the downregulation of the NF-kappa B signaling pathway.
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Alcaloides/farmacología , Antineoplásicos Fitogénicos/farmacología , Carcinoma Hepatocelular/tratamiento farmacológico , Inhibidores de la Metaloproteinasa de la Matriz , Quinolizinas/farmacología , Alcaloides/química , Alcaloides/uso terapéutico , Antineoplásicos Fitogénicos/uso terapéutico , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patología , Medicamentos Herbarios Chinos/química , Humanos , Metaloproteinasa 9 de la Matriz/genética , Estructura Molecular , FN-kappa B/antagonistas & inhibidores , FN-kappa B/genética , FN-kappa B/metabolismo , Quinolizinas/química , Quinolizinas/uso terapéutico , Sophora/química , MatrinasRESUMEN
AIM: To construct an eukaryotic expression vector containing the coding region of human full length cytokeratin 8 gene and to detect its expression in SMMC7721 cells. METHODS: CK8 cDNA was amplified by RT-PCR and cloned to pMD18-T simple vector. After confirming the sequence, the cDNA was inserted into pEGFP-C1 and the positive clone pEGFP-CK8 was obtained. The recombinant plasmid was transfected into SMMC7721 cells with Lipofectamine(TM);2000 and the expression was detected by fluorescence microscope, real time PCR and Western blot. The physical-chemical properties, signal peptide and functional motifs were predicted by the bioinformatics software. RESULTS: PCR, restriction enzyme digestion and DNA sequencing showed that the recombinant plasmid contained the coding region of full length CK8 gene. Observation under fluorescence microscope and the results of real time PCR and western blot indicated CK8 was over-expressed in SMMC7721 cells. CONCLUSION: The eukaryotic expression vector containing the CK8 gene was successfully constructed and expressed, which provides a basis for the study for biological function of CK8.
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Biología Computacional , Regulación de la Expresión Génica , Queratina-8/genética , Queratina-8/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Línea Celular Tumoral , Clonación Molecular , Vectores Genéticos/genética , Vectores Genéticos/metabolismo , Humanos , Queratina-8/química , TransfecciónRESUMEN
Several new clinical trials to compare the efficiency of pegylated interferon (PEGIFN) treatment with IFN treatment in Chinese patients with hepatitis B were published in recent years. However, the numbers of patients included in these clinical trials are too small to draw a clear conclusion whether PEGIFN is more effective than INF in Chinese patients. Therefore, a new meta-analysis was needed to be performed to compare PEGIFN with IFN in the treatment of Chinese hepatitis B patients. A search of MEDLINE, China National Knowledge Infrastructure, Wanfang Database and China Biomedical Database between 1966 and 2009 was performed. RCTs comparing the use of PEGIFN vs. IFN in treating Chinese patients with hepatitis B were assessed. Of the 210 studies screened, nine RCTs included 581 patients (PEGIFN therapy 281, IFN therapy 300) were analysed. The HBeAg seroconversion rates, serum HBeAg clearance rate, serum HBV DNA clearance rate and ALT normalization rate obtained in patients treated with PEGINF were significantly higher than in patients treated with IFN at the 24th, 48th week of the treatment and the 24th, 48th week of follow-up. Safety profile was similar between patients treated with PEGIFN and IFN. Chinese patients with hepatitis B have a greater likelihood of achieving a viral response, biomedical response and liver fibrosis improvement with PEGIFN α-2a.
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Antivirales/uso terapéutico , Hepatitis B/tratamiento farmacológico , Interferón-alfa/uso terapéutico , Polietilenglicoles/uso terapéutico , Antivirales/administración & dosificación , Biomarcadores/sangre , China , ADN Viral/sangre , Esquema de Medicación , Hepatitis B/sangre , Hepatitis B/complicaciones , Hepatitis B/epidemiología , Antígenos de la Hepatitis B/sangre , Humanos , Interferón alfa-2 , Interferón-alfa/administración & dosificación , Cirrosis Hepática/sangre , Cirrosis Hepática/epidemiología , Cirrosis Hepática/etiología , Pruebas de Función Hepática , Polietilenglicoles/administración & dosificación , Proteínas Recombinantes , Pruebas Serológicas , Resultado del TratamientoRESUMEN
OBJECTIVE: To investigate the expression of keratin 8 (K8) in carbon tetrachloride (CCl(4))-induced liver injury of mice. METHODS: Forty ICR mice were divided into four groups. CCl(4) 300 microl/kg body weight in olive oil was injected intraperitoneally for 0, 2, 4, 6 weeks in group A, B, C and D, respectively. Mice were sacrificed 3 d after the last injection and then the vital organs were collected and weighed. RT-PCR and immunohistochemistry methods were used to analyze the expression of keratin 8 in the liver. RESULTS: The ratios of liver and body weight were increased significantly after administration of CCl(4), which were 5.60 %, 6.87%, 7.83 % and 7.76% at 0, 2, 4 and 6 weeks after injection, respectively. The expression of K8 was increased at the 2 w, 4 w and 6 w after CL(4) administration. CONCLUSION: The expression of K8 is positively correlated with the liver injury induced by CCl(4). The accumulation of K8 may be involved in the mechanism of liver injury.
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Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Queratina-8/metabolismo , Animales , Tetracloruro de Carbono , Femenino , Queratina-8/genética , Masculino , Ratones , Ratones Endogámicos ICR , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
OBJECTIVE: To evaluate the therapeutic effect of acupuncture for the aphasia. METHODS: The publications of acupuncture for the treatment of aphasia are comprehensively searched from relevant domestic medical literature databases, China National Knowledge Infrastructure and Chinese BioMedical Literature Database. Meta analysis was conducted by using the publications. RESULTS: Twenty six trials of 338 matched the selection criteria and their data were suitable for Meta analysis. The total aphasia patients were 1749. The total odds ratio (OR) of the improvement with acupuncture plus language training and drugs compared with language training plus drug intervention was 3.66 (95% confidence interval, 2.81, 4.76), and the funnel plot was approximately symmetry. It is indicated that the curative effect of the acupuncture group is better than that of the control group (Z = 9.60, P < 0.001). CONCLUSION: The effect of acupuncture with language training plus drugs for the treatment of aphasia is better than that of language training plus drugs only.
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Terapia por Acupuntura , Afasia/terapia , Humanos , Terapia del Lenguaje , Ensayos Clínicos Controlados Aleatorios como AsuntoRESUMEN
BACKGROUND/AIMS: Resveratrol, a polyphenolic phytochemical present in berries, grapes, and wine, has emerged as a promising chemopreventive candidate. The aim of the present study was to determine the inhibitory effect of resveratrol on vascular endothelial growth factor (VEGF) expression and angiogenesis in hepatocellular carcinoma (HCC) and to explore its mechanism. METHODOLOGY: VEGF protein was detected by western blot, whereas VEGF mRNA expression was investigated by RT-PCR. Nuclear factor-kappa B (NF-kappa B) was measured by electrophoretic mobility shift assay (EMSA). Xenograft sections were stained for CD34 to study microvessels in vivo. RESULTS: We found that VEGF protein and mRNA expressions in the cells treated with resveratrol were significantly decreased. The activation of NF-kappa B was also intensely inhibited by resveratrol. Growth of tumours in nude mice was inhibited by resveratrol. Microvessel density was decreased with resveratrol treatment. CONCLUSIONS: The inhibitory effect of resveratrol on VEGF activity may occur partly through suppression of the activation of NF-kappa B in HepG2 cells. Resveratrol also significantly inhibited tumour growth and angiogenesis.
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Inhibidores de la Angiogénesis/farmacología , Anticarcinógenos/farmacología , Carcinoma Hepatocelular/tratamiento farmacológico , Neoplasias Hepáticas/tratamiento farmacológico , FN-kappa B/antagonistas & inhibidores , Estilbenos/farmacología , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Células Hep G2 , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , FN-kappa B/fisiología , ARN Mensajero/análisis , Resveratrol , Factor A de Crecimiento Endotelial Vascular/genéticaRESUMEN
OBJECTIVE: To perform a Meta-analysis on peginterferon with interferon in treatment of HIV patients coinfected with refractory genotype HCV. METHODS: A literature search of Medline was conducted to identify eligible randomized controlled trials. Meta analysis was conducted to evaluate peginterferon and interferon in treatment of coinfected HCV genotype 1 or 4 in HIV patients. RESULT: Six trials of 88 matched the selection criteria. Total 1,131 patients with coinfection of HCV genotype 1 or 4 and HIV were included. Sustain viral response was higher in patients treated with peginterferon plus ribavirin compared with that of interferon plus ribavirin (26 % compared with 8 %) or peginterferon alone (26 % compared with 13 %). Severe adverse effects and withdrawal rates were similar for patients treated with peginterferon and patients treated with interferon. CONCLUSION: Peginterferon plus ribavirin in treatment of patients with coinfection of genotype 1 or 4 HCV and HIV can achieve higher sustain viral response and the likelihoods of serious adverse effects and withdrawal rates are similar to other therapies.
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Infecciones por VIH/tratamiento farmacológico , Hepatitis C Crónica/tratamiento farmacológico , Interferón-alfa/administración & dosificación , Polietilenglicoles/administración & dosificación , Ribavirina/administración & dosificación , Adulto , Antivirales/administración & dosificación , Quimioterapia Combinada , Femenino , Genotipo , Infecciones por VIH/complicaciones , Infecciones por VIH/inmunología , Hepacivirus/clasificación , Hepacivirus/genética , Hepatitis C Crónica/complicaciones , Hepatitis C Crónica/virología , Humanos , Interferón alfa-2 , Masculino , Ensayos Clínicos Controlados Aleatorios como Asunto , Proteínas RecombinantesRESUMEN
The aim of this study was to assess whether PDSS2 (prenyl diphosphate synthase, subunit 2), a candidate tumor suppressor protein, has a potential anticancer role in human gastric cancer tissue and the SGC7901 gastric cell line. A PDSS2 eukaryotic expression vector was constructed and introduced into SGC7901 cells. The relationship between PDSS2 expression and cell proliferation, cell cycle distribution, and apoptosis in tumor cells was analyzed by RT-PCR, western blotting, the MTT colorimetric assay, flow cytometry, and immunohistochemistry. Increased exogenous PDSS2 expression in vitro is associated with decreased cellular proliferation of the gastric cancer cell line SGC7901. PDSS2 also induced apoptosis in SGC7901 cells by causing cell cycle arrest in the G0/G1 phase. Moreover, a significantly low expression level of PDSS2 protein was found in gastric cancer. Decreased or absent expression of PDSS2 was showed in the gastric tumor biopsy samples analyzed, correlating with cancer differentiation. PDSS2 has potent anticancer activity in gastric cancer tissues and the SGC7901 cell line and is possibly involved in apoptosis in SGC7901 cells.
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Transferasas Alquil y Aril/metabolismo , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Neoplasias Gástricas/enzimología , Apoptosis/genética , Ciclo Celular/genética , Línea Celular Tumoral , Proliferación Celular , Supervivencia Celular/genética , Humanos , TransfecciónRESUMEN
AIM: The present study examined the differential expression of proteins in HuH-7 cells and HuH-7 cells harboring in vitro-transcribed full-length hepatitis C virus 1b RNA (HuH-7-HCV), and elucidated the cellular responses to HCV replication. METHODS: The protein profiles of matched pairs of HuH-7-HCV cells and HuH-7 mock cells were analyzed by 2-D electrophoresis (2DE). Solubilized proteins were separated in the first dimension by isoelectric focusing, and by 12.5% SDS-PAGE in the second dimension. The differential protein expression was analyzed by use of image analysis software to identify candidates for HCV infection-associated proteins. RESULTS: In total, 29 protein spots showed increases and 25 protein spots showed decreases in signal in HuH-7-HCV cell 2DE profiles as compared with HuH-7 mock cells. In the next step, the 10 spots showing the greatest increase and the 10 spots showing the greatest decrease were excised from gels and the proteins present were identified by Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometer (MALDI-TOF MS) or MALDI-TOF/TOF MS. In total, 13 proteins were identified successfully. The potential significance of the differential expression due to HCV replication was discussed. CONCLUSION: Our study identifies changes in the proteome of HuH-7 cells in the presence of HCV replication and yields information of the mechanism of HCV pathogenesis. These results will be useful for the identification of HCV infection-associated proteins that could be molecular targets for treatment.
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Hepacivirus/genética , Hepacivirus/metabolismo , Proteómica , ARN Viral/biosíntesis , ARN Viral/genética , Línea Celular Tumoral , Humanos , Péptido Hidrolasas/química , Transfección , Proteínas Virales/química , Proteínas Virales/genética , Replicación ViralRESUMEN
OBJECTIVE: To investigate the effects of hepatitis C virus (HCV) on transcription regulation genes of host cells by gene chip assays in cultured cells with intact HCV genome. METHODS: Huh-7 hepatoma cells were cultured and infected with in vitro constructed HCV. The total RNAs, proteins and cell culture supernatants of HCV infected cells and control cells were isolated. Proteins and cell culture supernatants were used to detect the HCV replication and protein expression in cell culture system. The HCV protein expression was detected with Western blotting. Released HCV from infected cells was analyzed by real-time fluorescence quantitative PCR. Total RNA was qualified using 10 g/L agarose gel electrophoresis. cRNA was synthesized, fluorescence labeled and purified, then hybridized with Agilent oligo microarray (20173 probes). Differential expression of genes related to transcription in cell culture system was analyzed. RESULT: HCV was positive in cell culture supernatants and HCV protein expression was also positive according to Western blotting results. Eleven up-regulated and 11 down-regulated genes related to transcription were found after Agilent gene chip screening. CONCLUSION: Intact hepatitis C virus cell culture system provides an useful tool for study on the affects of HCV infection on transcription regulation genes in host cells.
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Regulación Neoplásica de la Expresión Génica , Genoma Viral , Hepacivirus/genética , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/virología , Línea Celular Tumoral , Perfilación de la Expresión Génica , Hepacivirus/crecimiento & desarrollo , Hepatocitos/virología , Humanos , Neoplasias Hepáticas/patología , Transcripción GenéticaRESUMEN
OBJECTIVE: To observe the effects of resveratrol on proliferation of human hepatocellular carcinoma cell line SMMC-7721 cells and expression of matrix metalloproteinase-9 (MMP-9) in vitro. METHODS: SMMC-7721 cells were treated with different concentrations of resveratrol for 24, 48 and 72 h, respectively. The effect of resveratrol on proliferation of SMMC-7721 cells was assessed with methyl thiazolyl tetrazolium (MTT). The expression of MMP-9 mRNA was determined by reverse transcription polymerase chain reaction (RT-PCR). MMP-9 protein was identified by Western blot analysis. RESULTS: Resveratrol could inhibit the proliferation of SMMC-7721 cells with dose- and time-dependent effects. Moreover, both MMP-9 mRNA expression and MMP-9 protein production were markedly reduced after resveratrol treatment. CONCLUSION: Resveratrol can inhibit the proliferation of SMMC-7721 cells and down-regulate MMP-9 expression. It is presumed that resveratrol may suppress the invasion and metastasis of hepatocellular carcinoma.
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Carcinoma Hepatocelular/enzimología , Neoplasias Hepáticas/enzimología , Metaloproteinasa 9 de la Matriz/metabolismo , Estilbenos/farmacología , Antineoplásicos Fitogénicos/farmacología , Carcinoma Hepatocelular/patología , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo/efectos de los fármacos , Humanos , Neoplasias Hepáticas/patología , Metaloproteinasa 9 de la Matriz/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Resveratrol , Células Tumorales CultivadasRESUMEN
Lack of proper study models has brought difficulties in the study of the mechanism of viral infection, life cycle and pathogenic mechanism of hepatitis C virus (HCV) and also become the major obstacles in development of efficient vaccine and new drugs for hepatitis C. In recent years, the establishment of robust HCV cell culture infection system and HCV transgenic animal provide powerful tools for the analysis of host virus interactions, which facilitate the discovery of antiviral drugs and vaccines for this important human pathogen.