RESUMO
In this study, the full-length complementary (c)DNA of common carp Cyprinus carpio melanoma differentiation-associated gene 5 (mda5) was cloned. The complete open reading frame of C. carpio mda5 contained 2982 bp and encodes 993 amino acids. The deduced amino acids contained six functional domains: two caspase activation and recruitment domains (CARD), a conserved restriction domain of bacterial type III restriction enzyme (ResIII), a DExD/H box-containing domain (DEXDc), a helicase super family C-terminal domain (HELICc) and a C-terminal regulatory domain (RD). The mda5 gene was expressed in all tested tissues, with high levels in the gills and spleen, while lower expressed in gonad and blood. The copy numbers of mda5 were increased in the liver, spleen, head kidney and the mucosal-associated immune tissues such as the foregut, hindgut, gills and skin after stimulation with polyinosinic polycytidylic [poly(I:C)] and Aeromonas hydrophila. The myxovirus resistance gene (mx) messenger (m)RNA levels in the spleen, head kidney, foregut and gills were significantly up-regulated after poly(I:C) injection. When injected with poly(I:C), mda5 and mx transcripts were also significantly induced in vitro. These results implied that mda5 might be involved in both antiviral and antibacterial innate immune processes in C. carpio. © 2016 The Authors. Journal of Fish Biology © 2016 The Fisheries Society of the British Isles.
Assuntos
Carpas/imunologia , Proteínas de Peixes/genética , Imunidade Inata , Helicase IFIH1 Induzida por Interferon/genética , Sequência de Aminoácidos , Animais , Carpas/microbiologia , Carpas/virologia , Clonagem Molecular , Proteínas de Peixes/metabolismo , Helicase IFIH1 Induzida por Interferon/metabolismo , Poli I-C/imunologia , RNA Mensageiro/metabolismo , Análise de Sequência de DNARESUMO
AIM: To analyze tanshinone IIA in Salvia miltiorrhiza Bunge by supercritical fluid extraction (SFE) coupled with capillary gas chromatography (CGC). METHODS: The solution pressure was predicted by solubility parameter theory, the extraction conditions were confirmed by orthogonal design in which temperature, volume of modifier and dynamic extracting volume had been studied in three levels. RESULTS: This method is quick, convenient, precise and accurate, added sample recovery is 95.3%, RSD = 4.27%. CONCLUSION: Compared with the method of ultrasonic extraction, significantly better qualitative and quantitative result could be obtained by SFE-CGC and expend less organic solvent then make less pollution.
Assuntos
Fenantrenos/análise , Plantas Medicinais/química , Salvia miltiorrhiza/química , Abietanos , Cromatografia Gasosa/métodos , Cromatografia com Fluido Supercrítico/métodosRESUMO
OBJECTIVE: To establish a gas chromatography fuzzy clustering analytical method for classifying of origin fungi of the Chinese traditional medicine, Hongqu. METHODS: The volatile and nonvolatile components of 7 ordinary species of Monascus including M. aurantiacus Lee, etc. were analyzed by capillary gas chromatog raphy using FID: Inlet temperature 280 degrees C, detector temperature 280 degrees C, temperature program 130 degrees C, 5 min-->10 degrees C.min-1-->200 degrees C, 10 min. RESULTS: In the peak number and contents of volatile and nonvolatile components of Monascus exist obviously differences through which Monascus can be distinguished easily by the main fingerprint peaks within 20 minutes. CONCLUSION: The capillary gas chromatography fuzzy clustering analytical method is useful to the identification of Monascus fungi.
