RESUMO
To increase the content of active constituent--RE and PD of Polygonum cuspidatum hairy root, through Ri-mediated gene transformation technology, modified high salt low pH method was used to distill genome DNA of grapevine (Vitis raparia). Primer was designed according to sequence of Genebank (AF128861). Through PCR amplification obtain RS gene sequence was obtained. Binary vector pCAMBIA1300-35S-RS was constructed. Frost thawing method was used to transform Agrobacterium rhizogenes ATCC11325. Scratched aseptic seedling leaf of Polygonum cuspidatum was contaminated subsequently. DNA conformity and mRNA expression of RS gene were investigated by PCR and RT-PCR respectively. RE and PD in transgenic hairy root were determined by HPLC. For the first time successfully inducement acquires transformed RS gene hairy root of Polygonum cuspidatum. Content of active constituents--RE and PD were 17 - 187 microg x g(-1) DW and 836 - 1 970 microg x g(-1) DW, respectively, the non-transgenic hairy root was 0 - 130 microg x g(-1) DW and 190 - 320 microg x g(-1) DW. In the different root selected, the content of PD was much higher than that in non-transformed hairy roots of Polygonum cuspidatum, the highest content is 5 times, but the content of RE has not increased apparently.
Assuntos
Aciltransferases , Genética , Metabolismo , Primers do DNA , DNA de Plantas , Genética , Medicamentos de Ervas Chinesas , Fallopia japonica , Genética , Metabolismo , Vetores Genéticos , Glucosídeos , Dados de Sequência Molecular , Raízes de Plantas , Genética , Metabolismo , Plantas Geneticamente Modificadas , Plantas Medicinais , Genética , Metabolismo , Rhizobium , Genética , Estilbenos , Transformação GenéticaRESUMO
To explore the relationship between T cell markers in hematological diseases and T cell markers in solid tumor, CD3, CD4, CD8 in hematological diseases, malignant and benign tumors were detected by flow cytometry and results were analyzed statistically. The results showed that CD3, CD4, CD8 and CD4/CD8 in chronic leukemia decreased significantly while these markers in acute leukemia and MDS decreased obviously in comparison with normal persons and other hematological diseases (P < 0.0l). Hemolytic anemia markers increased significantly (P < 0.05). CD3, CD4, CD4/CD8 in idiopathic thrombocytopenic purpura decreased and CD8 increased (P < 0.0l). CD3, CD4, CD8 in iron-deficiency anemia, anemia from chronic diseases, benign tumor and other hematological diseases were lower than those in normal persons and hemolytic anemia, but higher than those in acute and chronic leukemia, malignant tumor, granulocytopenia, and MDS (P > 0.05). It is notable that the above markers correlated with the development and prognosis of diseases. In conclusion, expression of CD3, CD4, CD8, CD4/CD8 contributes to diagnosis of hematological diseases and benign or malignant tumors, and is an important indicator for therapeutic strategy.