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At the urgent request of Coptis chinensis planting,growth suitability as assessment indicators for C. chinensis cultivation was proposed and analyzed in this paper , based on chemical quality determination and ecological fators analysis by Maxent and ArcGIS model. Its potential distribution areas at differernt suitability grade and regionalization map were formulated based on statistical theory and growth suitability theory. The results showed that the most suitable habitats is some parts of Chongqing and Hubei province, such as Shizhu, Lichuan, Wulong, Wuxi, Enshi. There are seven ecological factor is the main ecological factors affect the growth of Coptidis Rhizoma, including altitude, precipitation in February and September and the rise of precipitation and altitude is conducive to the accumulation of total alkaloid content in C. chinensis. Therefore, The results of the study not only illustrates the most suitable for the surroundings of Coptidis Rhizoma, also helpful to further research and practice of cultivation regionalization, wild resource monitoring and large-scale cultivation of traditional Chinese medicine plants.
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Coptis/crecimiento & desarrollo , Alcaloides/análisis , China , Coptis/química , Ecología , Ecosistema , Sistemas de Información Geográfica , Rizoma/química , Rizoma/crecimiento & desarrolloRESUMEN
CONTEXT: Lovastatin, a hypocholesterolemic drug, is produced by submerged fermentation of Aspergillus terreus Thom (Trichocomaceae). High performance liquid chromatography is usually used to determine lovastatin in samples of the fermentation broth. However, this method is inconvenient and costly, especially in the context of high-throughput sample analysis. OBJECTIVE: A direct and simple dual-wavelength ultraviolet spectrophotometric method for quantifying lovastatin in the fermentation broth of A. terreus was developed. MATERIALS AND METHODS: A. terreus Z15-7 was used for all experiments. The liquid fermentation was conducted at 30 °C in a rotary shaker at 150 rpm for 15 d. Silica gel and neutral alumina column chromatography were used for the separation and purification of lovastatin from the fermentation broth. RESULTS: The limits of detection of lovastatin were 0.320 µg/ml in the lovastatin standard solution and 0.490 µg/ml in the fermentation broth sample and the limits of quantification of lovastatin were 1.265 µg/ml in the lovastatin standard solution and 3.955 µg/ml in the fermentation broth sample. The amounts of lovastatin in the fermentation broth ranged from 876.614 to 911.967 µg/ml, with relative standard deviations from 1.203 to 1.709%. The mean recoveries of lovastatin using silica gel and neutral alumina column chromatography were 84.2 ± 0.82 and 87.2 ± 0.21%, respectively. DISCUSSION AND CONCLUSION: Dual-wavelength UV spectrophotometry is a rapid, sensitive, accurate, and convenient method for quantifying lovastatin in fermentation broth. Neutral alumina column chromatography is more efficient than silica gel column chromatography for the purification and determination lovastatin using the developed dual-wavelength UV spectrophotometry method.
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Anticolesterolemiantes/análisis , Aspergillus/metabolismo , Lovastatina/análisis , Espectrofotometría Ultravioleta/métodos , Cromatografía/métodos , Fermentación , Ensayos Analíticos de Alto Rendimiento/métodos , Límite de Detección , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: The purpose of this study was to evaluate the value of contrast-enhanced ultrasonography (CEUS) in differential diagnosis of superficial lymphadenopathy. METHODS: Ninety-four superficial enlarged lymph nodes in 94 patients were studied by conventional ultrasonography (gray scale and color Doppler) and CEUS. Contrast-enhanced sonograms were analyzed using contrast-specific quantification software. All of the results were compared with pathologic diagnoses. RESULTS: Of the 94 lymph nodes examined, 44 were benign and 50 were malignant (33 metastases and 17 lymphomas). The sensitivity, specificity, and accuracy of conventional ultrasonography in differential diagnosis between benign and malignant nodes were 51%, 47%, and 55%, respectively. Contrast-enhanced ultrasonography showed intense homogeneous enhancement in 39 of 44 benign lymph nodes, inhomogeneous enhancement in 32 of 33 metastases, and intense homogeneous enhancement and absence of perfusion in 9 of 17 and 6 of 17 lymphomas, respectively. The sensitivity specificity, and accuracy of CEUS were 84%, 79%, and 80%. After time-intensity curve gamma variates were calculated, the area under the curve of the benign lymph nodes was greater than those of the metastatic lymph nodes and lymphomas (P < .01). CONCLUSIONS: These results indicate that the use of CEUS and contrast-specific software has a higher degree of diagnostic accuracy than conventional ultrasonography for evaluations of superficial lymphadenopathy. The contrast enhancement patterns and time-intensity curves provide valuable diagnostic information for differential diagnosis of benign and malignant lymph nodes.
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Ganglios Linfáticos/diagnóstico por imagen , Enfermedades Linfáticas/diagnóstico por imagen , Fosfolípidos , Hexafluoruro de Azufre , Adolescente , Adulto , Anciano , Medios de Contraste , Femenino , Humanos , Persona de Mediana Edad , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Ultrasonografía , Adulto JovenRESUMEN
BACKGROUND: The cardiovascular characteristics of children with Hutchinson-Gilford progeria syndrome (HGPS) remain unclear. The present study is aimed at evaluating the cardiovascular changes with ultrasound examination in children with HGPS and compared these with those in normal children and older people. METHODS: Seven HGPS children, 21 age-matched healthy children, and 14 older healthy volunteers were evaluated by three-dimensional echocardiography (including strain analysis) and carotid elasticity examination with the echo-tracking technique. RESULTS: Children with HGPS had higher left ventricular ejection fraction (LVEF) and global longitudinal strain, when compared to older healthy volunteers (P < 0.05). However, these parameters were not significantly different, when compared to those in healthy children. Furthermore, children with HGPS had lower average peak times in the left ventricle, when compared with the other two groups. For the structure of the carotid artery detected by ultrasound, the abnormality rates were similar between children with HGPS and older healthy volunteers (83.3% vs. 71.4%). The elastic parameters, elastic modulus, stiffness parameter, and pulsed wave transmittal velocity of children with HGPS were lower, when compared to those in older healthy volunteers (P < 0.05), while they were higher with arterial compliance (P > 0.05). Furthermore, no significant difference existed among the vascular elastic parameters between HGPS and normal children. CONCLUSION: HGPS children had impaired left ventricular (LV) synchrony, when compared to normal children, although the difference in LVEF was not statistically significant. Furthermore, the structural abnormality of the carotid artery in HGPS children was similar to that in older people, although the index of elasticity appears to be more favorable. These results suggest that the cardiovascular system in HGPS children differs from natural aging.
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Envejecimiento , Arterias Carótidas , Diagnóstico por Imagen de Elasticidad , Ventrículos Cardíacos , Progeria , Análisis de la Onda del Pulso , Volumen Sistólico , Anciano , Arterias Carótidas/diagnóstico por imagen , Arterias Carótidas/fisiopatología , Niño , Preescolar , Femenino , Ventrículos Cardíacos/diagnóstico por imagen , Ventrículos Cardíacos/fisiopatología , Humanos , Lactante , Masculino , Persona de Mediana Edad , Progeria/diagnóstico por imagen , Progeria/fisiopatologíaRESUMEN
BACKGROUND/AIMS: To evaluate the antitumor immune response induced by truncated TERT (TERTt) mRNA transfected dendritic cells (DCs) in METHODOLOGY: Truncated mouse TERT sequence (according to mice telomerase reverse transcriptase mRNA 1776bp-2942bp) was cloned from B16 mice melanoma cells and inserted into pBluescript II KS(+) plasmid downstreaming of T7 promoter. The in vitro transcription was performed to prepare TERTt mRNA. The bone marrow-derived DCs isolated from BALB/c or C57B/L mice were electroporated with TERTt mRNA and recruited to immunize syngeneic naive mice respectively. The quantity and cytotoxic activity of tumor specific cytotoxic T lymphocytes (CTLs) in mice spleen were evaluated by using IFN-gamma enzyme-linked immunospot (ELIspot) and LDH release assay. The immunoprophylactic effects induced by TERTt mRNA transfected DC were evaluated in immunized-challenged mouse model. RESULTS: TERTt was cloned and transcripted into TERTt mRNA in vitro. TERTt mRNA transfected bone marrow-derived DCs were prepared. As shown by transfecting with EGFP mRNA, the DC transfected efficiency is 35.1% and there was a subtle increase of costimulator and MHC-II molecule expression after electroporation. Immunization with TERTt mRNA transfected DCs can induce TERTt and TERT-specific IFN-gamma secreting CTLs in the spleen of immunized mice. The splenocytes isolated from mice immunized with TERTt mRNA transfected DCs showed specific cytotoxic activity against TERTt and TERT-positive target cells. Using a syngeneic cancer mouse model, it was shown that TERTt mRNA transfected DCs vaccination can suppress the growth of TERT-positive tumor inoculation. CONCLUSIONS: TERTt mRNA transfected bone marrow-derived DCs can evoke antitumor immune response in vivo effectively and TERTt can serve as a universal tumor associated antigen to produce DC-based tumor vaccine.
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Vacunas contra el Cáncer/genética , Vacunas contra el Cáncer/inmunología , Células Dendríticas/inmunología , Células Dendríticas/trasplante , Neoplasias/terapia , Telomerasa/genética , Animales , Clonación Molecular , Células Dendríticas/enzimología , Electroporación , Femenino , Proteínas Fluorescentes Verdes/análisis , Proteínas Fluorescentes Verdes/genética , Ratones , Ratones Endogámicos , Trasplante de Neoplasias , ARN Mensajero/genética , Eliminación de Secuencia , Linfocitos T Citotóxicos/inmunología , Transfección , VacunaciónRESUMEN
Adenosine diphosphate P2Y12 receptor antagonist clopidogrel is not sufficiently safe for the gastric mucosa in patients with high risk of peptic ulcer, since it may impair healing of gastric erosions. However, the safety of the novel P2Y12 receptor antagonist ticagrelor in the gastric mucosa has not been elucidated to date. The present study aimed to examine whether ticagrelor delays gastric ulcer healing and to elucidate the involved mechanisms. Gastric kissing ulcers were produced in rats by luminal application of acetic acid solution, and ticagrelor was administered at dose of 10 or 20 mg/kg/day orally for 7 days. On day 8 after ulcer induction, the ulcer size, mucosal epithelial cell proliferation of the ulcer margin, expression levels of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF), and signal transduction pathways for cell proliferation and angiogenesis were measured and compared between the ticagrelor-treated and untreated model groups. The results revealed that the ulcer size was significantly greater in the ticagrelor-treated group compared with the model group, while the mucosal epithelial cell proliferation of the ulcer margin was significantly decreased in the ticagrelor-treated group. In addition, ticagrelor significantly decreased the ulcer-stimulated expression levels of EGF, VEGF, phosphorylated extracellular signal-regulated kinase (ERK), phosphorylated P38 mitogen-activated protein kinase and nuclear factor-κB P65 at the ulcer margin (P<0.05). These findings suggested that ticagrelor delayed gastric ulcer healing. Furthermore, the possible mechanisms underlying the effect of ticagrelor were associated with its functions of attenuating the expression levels of VEGF and EGF, as well as suppressing the phosphorylation activation of ERK1/2, P38 and nuclear factor-κB P65. Finally, the gastric epithelial cell proliferation and angiogenesis were also inhibited.
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AIM: To study whether heat-shocked tumor cells could enhance the effect of tumor cell lysate-pulsed dendritic cells (DCs) in evoking anti-tumor immune response in vivo. METHODS: Mouse undifferentiated colon cancer cells (CT-26) were heated at 42 degrees Celsius for 1 h and then frozen-thawed. The bone marrow-derived DCs pulsed with heat-shocked CT-26 cell lysate (HSCT-26 DCs) were recruited to immunize syngeneic naive BALB/c mice. The cytotoxic activity of tumor specific cytotoxic T lymphocytes (CTLs) in mouse spleen was evaluated by IFN-enzyme-linked immunospot (ELISpot) and LDH release assay. The immunoprophylactic effects induced by HSCT-26 DCs in mouse colon cancer model were compared to those induced by single CT-26 cell lysate-pulsed DCs (CT-26 DCs) on tumor volume, peritoneal metastasis and survival time of the mice. RESULTS: Heat-treated CT-26 cells showed a higher hsp70 protein expression. Heat-shocked CT-26 cell lysate pulsing elevated the co-stimulatory and MHC-II molecule expression of bone marrow-derived DCs as well as interleukin-12 p70 secretion. The IFN-gamma secreting CTLs induced by HSCT-26 DCs were significantly more than those induced by CT-26 DCs (P=0.002). The former CTLs' specific cytotoxic activity was higher than the latter CTLs' at a serial E/T ratio of 10:1, 20:1, and 40:1. Mouse colon cancer model showed that the tumor volume of HSCT-26 DC vaccination group was smaller than that of CT-26 DC vaccination group on tumor volume though there was no statistical difference between them (24 mm3 vs 8 mm3, P=0.480). The median survival time of mice immunized with HSCT-26 DCs was longer than that of those immunized with CT-26 DCs (57 d vs 43 d, P=0.0384). CONCLUSION: Heat-shocked tumor cell lysate-pulsed DCs can evoke anti-tumor immune response in vivo effectively and serve as a novel DC-based tumor vaccine.
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Antineoplásicos/inmunología , Extractos Celulares/inmunología , Línea Celular Tumoral , Células Dendríticas/inmunología , Calor , Animales , Células de la Médula Ósea/citología , Células de la Médula Ósea/inmunología , Neoplasias del Colon , Citocinas/inmunología , Células Dendríticas/citología , Femenino , Ratones , Ratones Endogámicos BALB C , Trasplante de Neoplasias , Tasa de SupervivenciaRESUMEN
Diketopiperazines are important secondary metabolites of the fungi with variety bioactivities. Several species belonging to genus Chaetomium produce compounds of this class, such as chetomin. To identify new antitumor agents, secondary metabolites of fungus Chaetomium sp 88194 were investigated and three new indole diketopiperazines, Chaetocochins G (1), Oidioperazines E (2) and Chetoseminudin E (3), along with two known compounds Chetoseminudins C (4) and N-acetyl-ß-oxotryptamine (5), were obtained. Chaetocochins G and Chetoseminudin E were recrystallized in CHCl3 containing a small amount of MeOH, and their structures with absolute configuration were established by spectroscopic data interpretation and single-crystal X-ray diffraction analysis. The absolute configuration of Oidioperazines E was defined by comparing of experimental and calculated electronic circular dichroism spectra. These isolates were also evaluated the anticancer activity, and Chaetocochins G displayed more potent cytotoxicity in MCF-7 cells than the common chemotherapeutic agent (5-fluorouracil) associated with G2/M cell cycle arrest. More importantly, Chaetocochins G induced cell apoptotic death via caspase-3 induction and proteolytic cleavage of poly (ADP-ribose) polymerase, concomitantly with increased Bax and decreased Bcl-2 expression. Our findings suggested that indole diketopiperazines from endophytic Chaetomium sp 88194 may be potential resource for developing anti-cancer reagents.
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Apoptosis/efectos de los fármacos , Chaetomium/metabolismo , Dicetopiperazinas/farmacología , Indoles/farmacología , Antineoplásicos/química , Antineoplásicos/farmacología , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Dicetopiperazinas/química , Expresión Génica , Humanos , Indoles/química , Células MCF-7 , Estructura Molecular , Resonancia Magnética Nuclear BiomolecularRESUMEN
Coumarin derivatives are an important class of C6-C3 plant metabolites that show a variety of bioactivities. Currently, most clinical anticoagulant agents are coumarins, such as warfarin, dicoumarol and acenocoumarol, and patients taking these drugs must be monitored for adverse reactions. In a search for safe and effective anticoagulant compounds from Chinese herbal medicine, a screening procedure on the whole plant of Ainsliaea fragrans was performed. The phytochemical investigation of this plant afforded five new coumarin derivatives, including a pair of natural 4-hydroxycoumarin enantiomers (1), a pair of coumarin enantiomers with a rare polycyclic pyrano[3-2c] carbon skeleton (2) and a 7-hydroxycoumarin derivative (3), together with 5 known biogenetically related compounds (4-8). Enantioseparation of 1 and 2 produced optically pure compounds 1a, 1b, 2a and 2b. The absolute configurations of the new compounds were confirmed by single-crystal X-ray diffraction analysis. In addition, we evaluated the anticoagulant activity of all isolates via activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT) assays in vitro and in vivo. Of note, compound 3 displayed potent anticoagulant activity and no significant hepatic or renal toxicity, which could make it a promising agent for further preclinical evaluation for preventing abnormal blood clotting.
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Anticoagulantes/aislamiento & purificación , Anticoagulantes/farmacología , Asteraceae/química , Cumarinas/aislamiento & purificación , Cumarinas/farmacología , Animales , Anticoagulantes/química , Coagulación Sanguínea/efectos de los fármacos , Espectroscopía de Resonancia Magnética con Carbono-13 , Dicroismo Circular , Cumarinas/química , Humanos , Espectroscopía de Protones por Resonancia Magnética , Ratas Wistar , EstereoisomerismoAsunto(s)
Síndrome de Budd-Chiari/cirugía , Hígado/cirugía , Hemorragia Posoperatoria/diagnóstico por imagen , Adulto , Angioplastia de Balón , Medios de Contraste , Humanos , Laparotomía , Hígado/diagnóstico por imagen , Masculino , Microburbujas , Hemorragia Posoperatoria/cirugía , Stents , Ultrasonografía Doppler en Color/métodosRESUMEN
Heavy-ion beams, possessing a wide mutation spectrum and increased mutation frequency, have been used effectively as a breeding method. In this study, the heavy-ion beams generated by the Heavy-Ion Research Facility in Lanzhou were used to mutagenize Aspergillus terreus CA99 for screening high-yield lovastatin strains. Furthermore, the main growth conditions as well as the influences of carbon and nitrogen sources on the growth and the lovastatin production of the mutant and the original strains were investigated comparatively. The spores of A. terreus CA99 were irradiated by 15, 20, 25, and 30 Gy of 80 MeV/u (12)C(6+) heavy-ion beams. Based on the lovastatin contents in the fermentation broth, a strain designated as A. terreus Z15-7 has been selected from the clone irradiated by the heavy-ion beam. When compared with the original strain, the content of lovastatin in the fermentation broth of A. terreus Z15-7 increased 4-fold. Moreover, A. terreus Z15-7 efficiently used the carbon and nitrogen sources for the growth and production of lovastatin when compared to the original strain. The maximum yield of lovastatin, 916.7 µg/ml, was obtained as A. terreus Z15-7 was submerged cultured in the chemically defined medium supplemented with 3% glycerol as a carbon source, 1% corn meal as an organic nitrogen source, and 0.2% sodium nitrate as an inorganic nitrogen source at 30 °C in the shake flask. The result shows that heavy-ion beam irradiation is an effective method for the mutation breeding of lovastatin production of A. terreus.
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Anticolesterolemiantes/metabolismo , Aspergillus/metabolismo , Carbono/metabolismo , Microbiología Industrial/métodos , Lovastatina/biosíntesis , Aspergillus/efectos de los fármacos , Aspergillus/genética , Aspergillus/crecimiento & desarrollo , Aspergillus/efectos de la radiación , Medios de Cultivo/metabolismo , Medios de Cultivo/farmacología , Fermentación , Glicerol/metabolismo , Iones Pesados , Transferencia Lineal de Energía , Lovastatina/metabolismo , Mutación , Nitratos/metabolismo , Nitrógeno/metabolismo , Radiación Ionizante , Espectrofotometría Infrarroja , Espectrofotometría Ultravioleta , Zea mays/metabolismoRESUMEN
OBJECTIVE: The purpose of this study was to investigate the feasibility of percutaneous microwave coagulation therapy (PMCT) guided by contrast-enhanced ultrasonography (CEUS) for controlling active bleeding in rabbit livers. METHODS: Twenty actively bleeding rabbit liver models, produced with an 18-gauge semiautomatic biopsy needle and confirmed with CEUS, were randomly divided into 2 groups: a PMCT group (n=10, with a microwave antenna placed into the bleeding site under ultra-sonographic guidance and worked at 60 W for 30 seconds on average) and a control group (n=10, with the active bleeding site not treated). After therapy procedures were performed, lactated Ringer's solution resuscitation was then performed in both groups to maintain the mean arterial pressure at 70 mm Hg for 1 hour. The intraperitoneal blood loss, total resuscitation volume, mean arterial pressure, and hematocrit value were recorded. Macroscopic and microscopic examinations were performed at the end of the study. RESULTS: After PMCT, the former bleeding site appeared on CEUS as a round or an oval area devoid of contrast. The PMCT group had lower blood loss (30.4+/-7.2 versus 101.6 +/- 18.2 mL; P< .05) and a lower total resuscitation volume (56.5+/-10 versus 186+/-36.6 mL; P< .05) than the control group. The mean hematocrit value in the PMCT group was significantly higher than that in the control group (26%+/-4% versus 19%+/-4%; P< .05) at the end of the experiment. CONCLUSIONS: Contrast-enhanced ultrasonographically guided PMCT significantly decreased blood loss in a rabbit model of active liver bleeding. It provides a simple and quick method to control blood loss in liver injuries with active bleeding.
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Hemorragia/terapia , Hígado/irrigación sanguínea , Microondas/uso terapéutico , Ultrasonografía Intervencional , Animales , Medios de Contraste , Estudios de Factibilidad , Hemorragia/diagnóstico por imagen , Hemostasis , Hígado/diagnóstico por imagen , Fosfolípidos , Conejos , Hexafluoruro de AzufreRESUMEN
MAGE-3, a member of melanoma antigen (MAGE) gene family, is recognized as an ideal candidate for tumor vaccine because it is expressed in a significant proportion of tumors of various histological types and can induce antigen-specific immune response in vivo. There is now substantial evidence that heat shock proteins HSPs isolated from cancer cells and virus-infected cells can be used as vaccines to produce cancer-specific or virus-specific immunity. In this research, we investigated whether M. tuberculosis HSP70 can be used as vehicle to elicit immune response to its accompanying MAGE-3 protein. A recombinant protein expression vector was constructed that permitted the production of fusion protein linking amino acids 195-314 of MAGE-3 to the C terminus of HSP70. We found that HSP70-MAGE-3 fusion protein can elicit stronger cellular and humoral immune responses against MAGE-3 expressing murine tumor than those elicited by MAGE-3 protein in vivo, which resulted in potent antitumor immunity against MAGE-3-expressing tumors. Covalent linkage of HSP70 to MAGE-3 was necessary to elicit immune response to MAGE-3. These results indicate that linkage of HSP70 to MAGE-3 enhanced immune responses to MAGE-3 in vivo and HSP70 can be exploited to enhance the cellular and humoral immune responses against any attached tumor-specific antigens.
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Antígenos de Neoplasias/inmunología , Vacunas contra el Cáncer/inmunología , Proteínas HSP70 de Choque Térmico/inmunología , Melanoma Experimental/terapia , Mycobacterium tuberculosis/inmunología , Proteínas de Neoplasias/inmunología , Linfocitos T Citotóxicos/inmunología , Vacunas de ADN/inmunología , Animales , Células Presentadoras de Antígenos , Antígenos de Neoplasias/genética , Femenino , Proteínas HSP70 de Choque Térmico/genética , Humanos , Inmunidad Celular , Melanoma Experimental/inmunología , Ratones , Ratones Endogámicos C57BL , Proteínas de Neoplasias/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Tasa de Supervivencia , Células Tumorales Cultivadas , VacunaciónRESUMEN
AIM: To construct the melanoma antigen-1(MAGE-1) eukaryotic expression plasmid and express MAGE-1 in mouse melanoma B16 cells. METHODS: The MAGE-1 gene was amplified by PCR and cloned into the eukaryotic expression vector pIRES2-EGFP to construct the pIRES2-EGFP-MAGE-1 plasmid. The plasmid was transfected into the B16 cells. The EGFP expression was detected under fluoroscent microscope and the MAGE-1 expression was detected by immunohistochemistry staining. RESULTS: The eukaryotic expression vector pIRES2-EGFP-MAGE-1 was constructed and transfected successfully into B16 cells, and the EGFP and MAGE-1 genes were co-expressed in the B16 cells. CONCLUSION: A mouse melanoma cell line B16 co-expressing MAGE-1 and EGFP genes has been established successfully, which lays the foundation for the research on application of MAGE-1 in the tumor immunotherapy.
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Línea Celular Tumoral , Proteínas Fluorescentes Verdes/biosíntesis , Melanoma Experimental/metabolismo , Proteínas de Neoplasias/biosíntesis , Animales , Antígenos de Neoplasias , Proteínas Fluorescentes Verdes/genética , Melanoma Experimental/patología , Antígenos Específicos del Melanoma , Ratones , Proteínas de Neoplasias/genética , Plásmidos , TransfecciónRESUMEN
The cancer-testis antigen encoded by the MAGE-1 gene is an attractive antigen in tumor immunotherapy because it can be processed as a foreign antigen by the immune system and generate tumor-specific cellular immune response in vivo. However, increase of the potency of MAGE-1 DNA vaccines is still needed. The high degree of sequence homology and intrinsic immunogenicity of heat shock protein 70 (HSP70) have prompted the suggestion that HSP70 might have immunotherapeutic potential, as HSP70 purified from malignant and virally infected cells can transfer and deliver antigenic peptides to antigen-presenting cells to elicit peptide-specific immunity. In this research, we evaluated the enhancement of linkage of Mycobacterium tuberculosis HSP70 to MAGE-1 gene of the potency of antigen-specific immunity elicited by naked DNA vaccines. We found that vaccines containing MAGE-1-HSP70 fusion genes enhanced the frequency of MAGE-1-specific cytotoxic T cells in contract to vaccines containing the MAGE-1 gene alone. More importantly, the fusion converted a less effective DNA vaccine into one with significant potency against established MAGE-1-expressing tumors. These results indicate that linkage of HSP70 to MAGE-1 gene may greatly enhance the potency of DNA vaccines, and generate specific antitumor immunity against MAGE-1-expressing tumors.