Summary: The specific anti-tumor immune response induced by mouse bone marrow dendritic cells (DCs) lransfected with recombinant adenovirus carrying mutant k-ras genes was investighted. DCs were generated from mous...Summary: The specific anti-tumor immune response induced by mouse bone marrow dendritic cells (DCs) lransfected with recombinant adenovirus carrying mutant k-ras genes was investighted. DCs were generated from mouse bone marrow in the presence of rmGM-CSF (3.3 ng/mL) and rmIL-4 (1.3 ng/mL) and detected by FACS, and then transfecled with the recombinant adenovirus encoding mutant k ras gene. The efficacy of transfection and T cell stimulating activity of DCs were detected. CTL activity of the mice vaccinated with DCs was observed. The resuhs showed thai DCs had dendritic veiled morphology. BmDCs highly expressed B7-1(80%), B7-2(77%), MHC Ⅱ (70%), CDllc (65%), CD40 (70%) and CD54 (96%) with FACS, and no significant difference in the expression was observed before and after the transfection (P〈0.05). The DCs transfeeled by mutant k-ras gene could significantly stimulate lymphoeytes proliferation as compared with those transfeeted by Ad e or non-modified DCs (P〈0.05). DC vaccine transfected by mutant k-ras gene could induce CTL activity against Lewis lung cancer, but not against B16. The specific eytotoxicity against Lewis lung cancer in Ad-k-ras/12-transdueed DC group was signifieantly higher than those in the control, vector and non transfeeted DCs groups (P〈0.05). It was concluded that special antitumor response could be induced by DCs transfected with recombinant adenovirus carrying mutant k-ras genes.展开更多
AIM To analyze sequences of the nuclear ribosomal RNA small subunit (18S rRNA) gene and the chloroplast mat K gene of crude drug Patchouli ( Pogostemon cablin ) in order to provide molecular evidence for identificatio...AIM To analyze sequences of the nuclear ribosomal RNA small subunit (18S rRNA) gene and the chloroplast mat K gene of crude drug Patchouli ( Pogostemon cablin ) in order to provide molecular evidence for identification of Patchouli drug. METHODS To sequence the entire 18S rRNA gene and partial mat K gene of Patchouli from Guangzhou and its substitute Wrinkled Gianthyssop ( Agastache rugosa ) from S ichuan using PCR direct sequencing and to detect the homology of two gene sequen ces between these two crude drugs. RESULTS The complete 18S rRNA gene sequence is 1 805 bp in length for Patchouli from Guangzhou whereas 1 794 bp for Wrinkled Gianthyssop from Sichuan. The 3′ end sequence of mat K gene is 521 bp (747~ 1 268 nt from upstream of ma t K gene) for these two crude drugs. Based on multiple sequence alignment, it i s found that there are 18 variable sites and 11 aligned gap sites in 18S rRNA se quence, 49 variable sites in 3′ mat K sequence between these two crude drug s. The homology is 98 4% for 18S rRNA and 90 6% for 3′ mat K between two crude drugs, respectively. CONCLUSION DNA sequencing can provide an accurate and reliable tool in the crude drug ident ification of Patchouli and its substitute Wrinkled Gianthyssop.展开更多
文摘Summary: The specific anti-tumor immune response induced by mouse bone marrow dendritic cells (DCs) lransfected with recombinant adenovirus carrying mutant k-ras genes was investighted. DCs were generated from mouse bone marrow in the presence of rmGM-CSF (3.3 ng/mL) and rmIL-4 (1.3 ng/mL) and detected by FACS, and then transfecled with the recombinant adenovirus encoding mutant k ras gene. The efficacy of transfection and T cell stimulating activity of DCs were detected. CTL activity of the mice vaccinated with DCs was observed. The resuhs showed thai DCs had dendritic veiled morphology. BmDCs highly expressed B7-1(80%), B7-2(77%), MHC Ⅱ (70%), CDllc (65%), CD40 (70%) and CD54 (96%) with FACS, and no significant difference in the expression was observed before and after the transfection (P〈0.05). The DCs transfeeled by mutant k-ras gene could significantly stimulate lymphoeytes proliferation as compared with those transfeeted by Ad e or non-modified DCs (P〈0.05). DC vaccine transfected by mutant k-ras gene could induce CTL activity against Lewis lung cancer, but not against B16. The specific eytotoxicity against Lewis lung cancer in Ad-k-ras/12-transdueed DC group was signifieantly higher than those in the control, vector and non transfeeted DCs groups (P〈0.05). It was concluded that special antitumor response could be induced by DCs transfected with recombinant adenovirus carrying mutant k-ras genes.
文摘AIM To analyze sequences of the nuclear ribosomal RNA small subunit (18S rRNA) gene and the chloroplast mat K gene of crude drug Patchouli ( Pogostemon cablin ) in order to provide molecular evidence for identification of Patchouli drug. METHODS To sequence the entire 18S rRNA gene and partial mat K gene of Patchouli from Guangzhou and its substitute Wrinkled Gianthyssop ( Agastache rugosa ) from S ichuan using PCR direct sequencing and to detect the homology of two gene sequen ces between these two crude drugs. RESULTS The complete 18S rRNA gene sequence is 1 805 bp in length for Patchouli from Guangzhou whereas 1 794 bp for Wrinkled Gianthyssop from Sichuan. The 3′ end sequence of mat K gene is 521 bp (747~ 1 268 nt from upstream of ma t K gene) for these two crude drugs. Based on multiple sequence alignment, it i s found that there are 18 variable sites and 11 aligned gap sites in 18S rRNA se quence, 49 variable sites in 3′ mat K sequence between these two crude drug s. The homology is 98 4% for 18S rRNA and 90 6% for 3′ mat K between two crude drugs, respectively. CONCLUSION DNA sequencing can provide an accurate and reliable tool in the crude drug ident ification of Patchouli and its substitute Wrinkled Gianthyssop.