RESUMEN
1-deoxy-D-xylulose-5-phosphate synthase2(DXS2) is the first key enzyme of the MEP pathway,which plays an important role in terpene biosynthesis of plants. According to the data of Swertia mussotii transcriptome, DXS2 gene(Gen Bank number MH535905) was cloned and named as Sm DXS2. The bioinformatics results showed that Sm DXS2 has no intron,with a 2 145 bp open reading frame encoding a polypeptide of 714 amino acids. They are belonging to 20 kinds of amino acids,and the most abundant amino acids include Ala,Gly and Trp. The predicted protein molecular weight was 76. 91 k Da and its theoretical isoelectric point(p I) was6. 5,which belonging to a hydrophilic protein. α-Helix and loop were the major motifs of predicted secondary structure of DXS2. The three function domains are TPP_superfamily,Transket_pyr_ superfamily and Transketolase_C superfamily,respectively. The Sm DXS2 protein shared high identity with other DXS2 proteins of plants. Phylogenetic analysis showed that Sm DXS2 protein is grouped with the gentian DXS2 protein. The recombinant protein of Sm DXS2 gene in Escherichia coli was approximately 92. 00 k Da(containing sumo-His tag protein 13 k Da),which was consistent with the anticipated size.This work will provide a foundation for further functional research of Sm DXS2 protein and increasing the product of iridoid compound by genetic engineering in S. mussotii.
Asunto(s)
Proteínas de Plantas/genética , Swertia/genética , Transferasas/genética , Secuencia de Aminoácidos , Clonación Molecular , ADN Complementario/genética , Genes de Plantas , Iridoides , Filogenia , Swertia/enzimología , TranscriptomaRESUMEN
The endophytic fungi with plant growth promoting effects were screened by co-culture of each endophytic fungus and seedlings of Dendrobium officinale. Anatomical features of the inoculated roots were studied by paraffin sectioning. Morphological characteristics and rDNA ITS1-5. 8S-ITS2 sequences were applied for the taxonomy of endophytic fungi. The results showed that 8 strains inoculated to D. officinale seedlings greatly enhanced plant height, stem diameter, new roots number and biomass. According to the anatomical features of the inoculated roots, each fungus could infect the velamina of seedlings. The hyphae or pelotons were existed in the exodermis passage cells and cortex cells. The effective fungi could not infect the endodermis and vascular bundle sheath, but which was exception for other fungi with harmful to seedlings. Combined with classic morphologic classification, 2 effective strains were identified which were subjected to Pestalotiopsis and Eurotium. Six species of fungi without conidiophore belonged to Pyrenochaeta, Coprinellus, Pholiota, Alternaria, Helotiales, which were identified by sequencing the PCR-amplified rDNA ITS1-5. 8S-ITS2 regions. The co-culture technology of effective endophytic fungi and plant can apply to cultivate the seedlings of D. officinale. It is feasible to shorten growth cycle of D. officinale and increase the resource of Chinese herbs.
Asunto(s)
Dendrobium/crecimiento & desarrollo , Dendrobium/microbiología , Endófitos/fisiología , Hongos/fisiología , Biomasa , ADN de Hongos/genética , ADN Ribosómico/genética , ADN Espaciador Ribosómico/genética , Dendrobium/citología , Endófitos/clasificación , Endófitos/genética , Hongos/clasificación , Hongos/genética , Interacciones Huésped-Patógeno , Hifa/fisiología , Raíces de Plantas/citología , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/microbiología , Plantas Medicinales/crecimiento & desarrollo , Plantas Medicinales/microbiología , ARN Ribosómico 5.8S/genética , Plantones/citología , Plantones/crecimiento & desarrollo , Plantones/microbiología , Especificidad de la EspecieRESUMEN
Interactions between an isolate of dark septate endophytes (DSE) and roots of Dendrobium nobile Lindl. seedlings are reported in this paper. The isolate was obtained from orchid mycorrhizas on Dendrobium sp. in subtropical forest. The fungus formed typical orchid mycorrhiza in aseptic co-culture with D. nobile seedlings on modified Murashige-Skoog (MMS) medium. Anatomic observations of the infected roots showed that the DSE hyphae invaded the velamen layer, passed through passage cells in exodermis, entered the cortex cells, and then formed fungal pelotons of orchid mycorrhiza. D. nobile seedlings' plant height, stem diameter, new roots number and biomass were greatly enhanced by inoculating the fungus to seedlings. The fungus was identified as Leptodontidium by sequencing the polymerase chain reaction-amplified rDNA ITS1-5.8S-ITS2 (internal transcribed spacer (ITS)) regions and comparison with similar taxa.
Asunto(s)
Dendrobium/microbiología , Hongos/aislamiento & purificación , Interacciones Huésped-Patógeno , Raíces de Plantas/microbiología , Plantones/microbiología , Medios de Cultivo/farmacología , ADN de Hongos/genética , ADN Intergénico/genética , ADN Ribosómico/genética , Dendrobium/efectos de los fármacos , Dendrobium/crecimiento & desarrollo , Hongos/citología , Hongos/genética , Interacciones Huésped-Patógeno/efectos de los fármacos , Filogenia , Raíces de Plantas/efectos de los fármacos , Plantones/efectos de los fármacos , Plantones/genéticaRESUMEN
OBJECTIVE: The objective of the present study is to investigate the inhibitory effects of sinomenine (SIN) on angiogenesis in a collagen-induced arthritis (CIA) mouse model. METHODS: Arthritis assessments for all mice were recorded. The histopathological assessments were performed following haematoxylin and eosin (HE) staining. Immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) analyses were used to detect the expression of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and angiopoietin 1 (ANG-1) in the serum and in the membrane. Immunohistochemistry was employed to detect the synovium microvessel density (MVD). RESULTS: Compared with the CIA model group, SIN significantly ameliorated swelling and erythema extension, decreased the arthritis index, reduced inflammation, cartilage damage and bone erosion, and lessened the number of CD31 positive cells on the synovium. Moreover, the levels of HIF-1α, VEGF and ANG-1 in the synovium and in the peripheral serum were increased in the untreated CIA model group but were significantly reduced in the 30 mg/kg, 100 mg/kg and 300 mg/kg SIN treatment groups. CONCLUSION: SIN could mitigate CIA by inhibiting angiogenesis, and the mechanism may associate with the HIF-1α-VEGF-ANG-1 axis. Additionally, our study provides a referable experimental basis for the use of SIN for the treatment of rheumatoid arthritis.
Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Artritis Experimental/tratamiento farmacológico , Morfinanos/farmacología , Neovascularización Patológica/tratamiento farmacológico , Angiopoyetina 1/metabolismo , Animales , Antirreumáticos/farmacología , Artritis Experimental/fisiopatología , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/fisiopatología , Colágeno/toxicidad , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Ratones , Ratones Endogámicos C57BL , Neovascularización Patológica/patología , Membrana Sinovial/patología , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
BACKGROUND: Clinical outcomes of undifferentiated arthritis (UA) are diverse, and only 40% of patients with UA develop rheumatoid arthritis (RA) after 3 years. Discovering predictive markers at disease onset for further intervention is critical. Therefore, our objective was to analyze the clinical outcomes of UA and ascertain the predictors for RA development. METHODS: We performed a prospective, multi-center study from January 2013 to October 2016 among Chinese patients diagnosed with UA in 22 tertiary-care hospitals. Clinical and serological parameters were obtained at recruitment. Follow-up was undertaken in all patients every 12 weeks for 2 years. Predictive factors of disease progression were identified using multivariate Cox proportional hazards regression. RESULTS: A total of 234 patients were recruited in this study, and 17 (7.3%) patients failed to follow up during the study. Among the 217 patients who completed the study, 83 (38.2%) patients went into remission. UA patients who developed RA had a higher rheumatoid factor (RF)-positivity (42.9% vs. 16.8%, χâ=â8.228, Pâ=â0.008), anti-cyclic citrullinated peptide (CCP) antibody-positivity (66.7% vs. 10.7%, χâ=â43.897, Pâ<â0.001), and double-positivity rate of RF and anti-CCP antibody (38.1% vs. 4.1%, χâ=â32.131, Pâ<â0.001) than those who did not. Anti-CCP antibody but not RF was an independent predictor for RA development (hazard ratio 18.017, 95% confidence interval: 5.803-55.938; Pâ<â0.001). CONCLUSION: As an independent predictor of RA, anti-CCP antibody should be tested at disease onset in all patients with UA.
Asunto(s)
Artritis Reumatoide/etiología , Artritis/complicaciones , Autoanticuerpos/sangre , Péptidos Cíclicos/inmunología , Adulto , Artritis/inmunología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Modelos de Riesgos Proporcionales , Estudios ProspectivosRESUMEN
C57BL/6 mice were inoculated intranasally (50 microl) with serial 10-fold dilution of HAB/01 H5N1 virus. Three and five days later, three mice of each group were euthanized. Lung injury was assessed by observation of lung histopathology, virus titers and MCD50 were also measured. Our data showed that H5N1 viral infection in mice resulted in mainly epithelial injury and interstitial pneumonia, featuring significant weight loss, dramatically increased lung wet weight:body weight ratio, inflammatory cellular infiltration, alveolar and interstitial edema, hemorrhage in lungs with high virus titers, and MCD50 was 10(-6.5)/ 0.05 mL. These results suggested that a mouse model of H5N1 viral infection was successfully established which may benefit study of H5N1 avian influenza virus and pathogenic mechanism of host.