RESUMO
The enhanced green fluorescent protein (EGFP) pEGFP-N1-P53 eukaryotic expression vector, which contains the human tumor suppressor p53, was constructed and transfected into chicken fibroblast cells and stage-X blastoderm to analyze the transfection efficiency. The complementary DNA of the human p53 gene was cloned by reverse transcription-polymerase chain reaction from human peripheral blood and inserted into the pEGFP-N1 vector by HindIII and BamHI double digestion. The pEGFP-N1-P53 vector was transfected into chicken embryo fibroblasts by Lipofectamine 2000 liposomes, and the transfection efficiency was analyzed by fluorescence microscope after 36 h of transfection. The stage-X blastoderm was also transfected by blastoderm injection using Lipofectamine 2000 liposomes at room temperature after 12-24 h; then hatching occurred until seventh day, and the transfection efficiency was analyzed by fluorescence microscope in the dead embryo. A total of 90 hatching eggs were transfected by the pEGFP-N1-P53 vector, and 20 chicken embryos expressed the reporter gene, which indicated that recombinant pEGFP-N1-P53 could be transfected and expressed in stage-X blastoderm by liposomes. Chicken embryo fibroblasts were transfected and expressed the reporter gene. The pEGFP-N1-P53 vector was constructed successfully and could be transfected and expressed in chicken embryo fibroblasts and stage-X blastoderms efficiently.
Assuntos
Vetores Genéticos , Proteínas de Fluorescência Verde/genética , Proteínas Recombinantes de Fusão/genética , Proteína Supressora de Tumor p53/genética , Animais , Blastoderma/crescimento & desenvolvimento , Blastoderma/metabolismo , Embrião de Galinha , Fibroblastos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Proteínas Recombinantes de Fusão/biossíntese , Proteína Supressora de Tumor p53/biossínteseRESUMO
OBJECTIVE: To observe the effect of scalp acupuncture on focal cerebral ischemia in rats. METHODS: Focal cerebral ischemia was produced in rats by the occlusion of right middle cerebral artery. The therapeutic action of scalp acupuncture on focal cerebral ischemia was studied by investigating the change of weight, neurologic symptoms, passive conditional reflex, hemorheology, infarct size of brain and pathology of brain tissue. RESULTS: Scalp acupuncture could not only improve obviously neurologic symptoms, prolong latent period of the passive conditional reflex and lower blood viscosity, but also lessen infarct size, promote proliferation and repair of neogenetic capillary and gliocyte in the malactic and necrotic regions and reduce the edema and inflammatory reaction around the necrotic region. CONCLUSION: Scalp acupuncture have therapeutic action on acute focal cerebral ischemia.
