用硅胶柱色谱及重结晶等方法,并根据理化性质和光谱数据从一株红树林共生真菌Paecilomycessp.Tree 1-7中分离鉴定了四个化合物,分别为secalon ic ac id A(1),tenellic ac id A(2),大黄素(3),大黄酚(4)。并首次对化合物1和2进行了抗人的...用硅胶柱色谱及重结晶等方法,并根据理化性质和光谱数据从一株红树林共生真菌Paecilomycessp.Tree 1-7中分离鉴定了四个化合物,分别为secalon ic ac id A(1),tenellic ac id A(2),大黄素(3),大黄酚(4)。并首次对化合物1和2进行了抗人的肝癌细胞hepG2的活性测试。结果显示,化合物1有很强的抗肿瘤活性,IC50=2.0μg/m l,化合物2的抗肿瘤活性IC50=62.1μg/m l。展开更多
AIM: To study the activation of cytotoxic T lymphocytes (CTLs) against gastric cancer cells induced by FasL/B7-1 (FB-11) gene-modified tumor cells, and to explore whether co-expression of FasL and B7-1 in SGC-7901 tum...AIM: To study the activation of cytotoxic T lymphocytes (CTLs) against gastric cancer cells induced by FasL/B7-1 (FB-11) gene-modified tumor cells, and to explore whether co-expression of FasL and B7-1 in SGC-7901 tumor cells could initiate synergistic antitumor effect. METHODS: FasL and B7-1 genes were transfected into human SGC-7901 gastric cancer cells with adenovirus vectors. The positive clones were selected by G418. FasL and B7-1 genes were detected by flow cytometry and RT-PCR. Abdominal infiltrating lymphocytes and sensitized spleen cells were obtained from mice that were immunized with SGC-7901/FB-11 or wild type SGC-7901 cells intraperitoneally, and cytotoxicity of these CTLs against tumor cells was determined by MTT assay. RESULTS: Flow cytometry and RT-PCR showed that FasL and B7-1 genes were highly expressed. FasL and B7-1 transfected cancer cells had a high apoptosis index. DNA laddering suggested that FasL and B7-1 genes induced gastric cancer cell apoptosis. FasL+/B7-1+SGC-7901 cells (SGC-7901/FB-11) were inoculated subcutaneously in the dorsal skin of C57BL/6 mice and then decreased their tumorigenicity greatly (z = 2.15-46.10, P＜0.01). SGC- 7901/FB-11 cell-sensitized mice obtained protective immune activity against the rechallenge of wild type SGC 7901 cells (z = 2.06-44.30, P＜0.05). The cytotoxicity of CTLs induced by SGC-7901/FB-11 cells against SGC-7901 was significantly higher than that of CTLs activated by wild-type SGC-7901 cells (84.1±2.4% vs30.5±2.3%,P＜0.05).CONCLUSION: FasL and B7-1 genes can effectively promote the activity of CTLs against gastric cancer cells. FasL/B7-1 molecules play an important role in CTL cytotoxicity.展开更多
Objective： To construct eukaryotic expression vector containing B7-1/GFP geneand study its expression in osteosarcoma cell line LM8. Methods： By using gene cloning technique, eukaxyotic expression vector pEGFP-C1 wa...Objective： To construct eukaryotic expression vector containing B7-1/GFP geneand study its expression in osteosarcoma cell line LM8. Methods： By using gene cloning technique, eukaxyotic expression vector pEGFP-C1 was used to construct the murine B7-1 recombinant plasmid （pEGFP-C1/B7）. Recombinant plasmid was transfected into LM8 cells with liposome and was confirmed by restriction endonuclease digestion and DNA sequencing. The expression of the fusion protein was detected using fluorescence microscope and Western blot analysis. Results： The recombinant eukaryotic expression plasmid pEGFP-C1/B7 was successfully constructed, which was confirmed by DNA sequencing, RT-PGR and restriction enzymes analysis. The green fluorescent protein could be detected in the transfected LM8 with fluorescence microscope. The expected B7-1 and green fluorescent protein （GFP） fusion protein was detected by RT-PCR and Western blot. Conclusion： The eukaryotic expression vector containing B7-1/GFP gene was constructed successfully, and it could be expressed in LM8 after transfection.展开更多
Objective: To study the vaccine potency of gene-modified tumor cells. Methods: The EL-4 lymphoma was transduced with recombinant retrovirus containing the murine GM-CSF gene or B7-1 gene. The effect of gene transducti...Objective: To study the vaccine potency of gene-modified tumor cells. Methods: The EL-4 lymphoma was transduced with recombinant retrovirus containing the murine GM-CSF gene or B7-1 gene. The effect of gene transduction on antitumor immunity was investigated. Results: Flow cytometry analysis showed that expression of their surface marker between wild-type EL-4 cells and gene transduced tumor cells was the same except for CD80 positive in B7-1 gene transduced cells. GM-CSF gene or B7-1 gene transduced EL-4 cells resulted in remarkable loss of tumorigenicity in syngenetic mice. The systemic protective immunity was induced against the challenge with EL-4/wt cells. Therapeutic vaccine with EL-4/GM-CSF or EL/7-1 cells could retard the growth of established early-stage EL-4/wt tumor significantly, but not retard the growth of late-stage EL-4/wt tumor. Irradiated GM-CSF gene transduced EL-4 cells showed strong vaccine effect against EL-4 cell challenge, but irradiated B7-1 gene transduced EL-4 cells showed weak vaccine effect. Remarkable cooperative antitumor effect against EL-4 cell challenge was observed when both irradiated EL-4/GM-CSF and EL-4/B7-1 were inoculated together. Conclusion: GM-CSF gene or B7-1 gene transduced BL-4 cells can be used as a good tumor vaccine. The combination of the two kinds of vaccine may have potential application value in human cancer treatment.展开更多
Objective: To construct recombinant R7-l/B7-2 retrovirus vectors and observe the effects of B7-l/R7-2 gene expression on in ho and in for immune response against against murine hepatoma. Methods: The recombinant retro...Objective: To construct recombinant R7-l/B7-2 retrovirus vectors and observe the effects of B7-l/R7-2 gene expression on in ho and in for immune response against against murine hepatoma. Methods: The recombinant retrovirus vectors expressing B7-1/B7-2 were constructed by gene cloning technology to produce retrovirus-infected PE501 and PA317 cell lines and murine hepatoma Hepal-6. The expression of R7-l/B7-2 was detected by fluorescence activated cell soning analysis (FACS). B7-l/B7-2 positive Hepal-6 Cell lines were used in inducing anti-hepatoma immunity in ho and in the. Results: In contrast to the excessive growth of parental Hemal-6 tumor, the growth of B7-l/B7-2-positive Hepal-6 inoculated into syngenic mice regressed. B7-1/R7-2-positive or cytokine-treated Hepal-6 alone could only induce mild cytototicity; in contrast, B7-1/B7-2-positive Hemal-6 treated with cytokine-stimulated spleen cells and activated the cytotoxicity effectively. Immunity in mice with R7-1/B7-2-positive tumor cells or cytokine-beated Hepal-6 only provided partial protection against parental Hepa1-6 tumor, whereas pretreatment of the transfected tumor cells with IFN-r and TNF-a induced complete immunity protection in vivo. Mice receiving inoculation of cytokine-treated B7-l/R7-2-positive Hemal-6 cells presented regression of the establoshed pental tUmor and survived for more than l00 d, while those untreated mice died within 40 d. Conclu sions: B7-l/R7-2 expression is necessary but not sufficient in inducing anti-hepatoma immune response, whereas it is efficient when combined with the beatment of IFN-γ and TNF-a.展开更多
A new degraded sesquiterpene was isolated from the marine actinomycete Streptomyces sp. 0616208. Its structure was elucidated as （1α; 4aα; 5α, 7β, 8aβ）-5, 8a-dimethyl-decahydrona- phthalene-1, 4a, 7-triol on th...A new degraded sesquiterpene was isolated from the marine actinomycete Streptomyces sp. 0616208. Its structure was elucidated as （1α; 4aα; 5α, 7β, 8aβ）-5, 8a-dimethyl-decahydrona- phthalene-1, 4a, 7-triol on the basis of spectroscopic data.展开更多
目的 对从混合培养的海洋软珊瑚共附生真菌中获得的结构新颖、活性良好的杂萜类成分进行研究。方法采用大米培养基对2株软珊瑚共附生曲霉属真菌EGF7-0-1和EGF15-0-3进行共培养;根据杂萜类化合物的结构特征,在基于质谱的分子网络(GNPS)...目的 对从混合培养的海洋软珊瑚共附生真菌中获得的结构新颖、活性良好的杂萜类成分进行研究。方法采用大米培养基对2株软珊瑚共附生曲霉属真菌EGF7-0-1和EGF15-0-3进行共培养;根据杂萜类化合物的结构特征,在基于质谱的分子网络(GNPS)和薄层色谱(TLC)的共同指导下,运用多种柱色谱及高效液相色谱(HPLC)等方法对其杂萜类成分进行目标导向分离;通过核磁共振(NMR)、高分辨质谱(HRMS)和旋光(ORD)等技术及物理常数对照等手段对单体化合物进行结构鉴定;采用液相色谱质谱联用(LC-MS^(n))和GNPS技术分析比较获得的杂萜类成分在2株真菌共培养及EGF7-0-1单培养中的含量。结果 从2株海洋真菌Aspergillus sp. EGF7-0-1和EGF15-0-3共培养的大米培养基中共分离得到7个杂萜类化合物,结构分别为terretonin (1)、terretonin A (2)、terretonin D1 (3)、terretonin H (4)、terreustoxin E (5)、aperterpene N (6)和asperterpene J (7)。LC-MSn和GNPS分析结果表明共培养条件下杂萜类化合物更为丰富。结论 化合物1~7均为具6/6/6骈并骨架的3,5-二甲基苔色酸(DMOA)途径衍生而成的杂萜。与菌株EGF7-0-1单培养相比,2株真菌共培养可以诱导产生丰富的杂萜,为进一步新颖杂萜类化合物的挖掘提供支持数据。展开更多
文摘用硅胶柱色谱及重结晶等方法,并根据理化性质和光谱数据从一株红树林共生真菌Paecilomycessp.Tree 1-7中分离鉴定了四个化合物,分别为secalon ic ac id A(1),tenellic ac id A(2),大黄素(3),大黄酚(4)。并首次对化合物1和2进行了抗人的肝癌细胞hepG2的活性测试。结果显示,化合物1有很强的抗肿瘤活性,IC50=2.0μg/m l,化合物2的抗肿瘤活性IC50=62.1μg/m l。
基金Supported by the Medical Scientific Foundation of Jiangsu Province, No. H200147
文摘AIM: To study the activation of cytotoxic T lymphocytes (CTLs) against gastric cancer cells induced by FasL/B7-1 (FB-11) gene-modified tumor cells, and to explore whether co-expression of FasL and B7-1 in SGC-7901 tumor cells could initiate synergistic antitumor effect. METHODS: FasL and B7-1 genes were transfected into human SGC-7901 gastric cancer cells with adenovirus vectors. The positive clones were selected by G418. FasL and B7-1 genes were detected by flow cytometry and RT-PCR. Abdominal infiltrating lymphocytes and sensitized spleen cells were obtained from mice that were immunized with SGC-7901/FB-11 or wild type SGC-7901 cells intraperitoneally, and cytotoxicity of these CTLs against tumor cells was determined by MTT assay. RESULTS: Flow cytometry and RT-PCR showed that FasL and B7-1 genes were highly expressed. FasL and B7-1 transfected cancer cells had a high apoptosis index. DNA laddering suggested that FasL and B7-1 genes induced gastric cancer cell apoptosis. FasL+/B7-1+SGC-7901 cells (SGC-7901/FB-11) were inoculated subcutaneously in the dorsal skin of C57BL/6 mice and then decreased their tumorigenicity greatly (z = 2.15-46.10, P＜0.01). SGC- 7901/FB-11 cell-sensitized mice obtained protective immune activity against the rechallenge of wild type SGC 7901 cells (z = 2.06-44.30, P＜0.05). The cytotoxicity of CTLs induced by SGC-7901/FB-11 cells against SGC-7901 was significantly higher than that of CTLs activated by wild-type SGC-7901 cells (84.1±2.4% vs30.5±2.3%,P＜0.05).CONCLUSION: FasL and B7-1 genes can effectively promote the activity of CTLs against gastric cancer cells. FasL/B7-1 molecules play an important role in CTL cytotoxicity.
基金This study was supported by a grant from the youth dawn program of Wuhan (Grant No. 20025001028).
文摘Objective： To construct eukaryotic expression vector containing B7-1/GFP geneand study its expression in osteosarcoma cell line LM8. Methods： By using gene cloning technique, eukaxyotic expression vector pEGFP-C1 was used to construct the murine B7-1 recombinant plasmid （pEGFP-C1/B7）. Recombinant plasmid was transfected into LM8 cells with liposome and was confirmed by restriction endonuclease digestion and DNA sequencing. The expression of the fusion protein was detected using fluorescence microscope and Western blot analysis. Results： The recombinant eukaryotic expression plasmid pEGFP-C1/B7 was successfully constructed, which was confirmed by DNA sequencing, RT-PGR and restriction enzymes analysis. The green fluorescent protein could be detected in the transfected LM8 with fluorescence microscope. The expected B7-1 and green fluorescent protein （GFP） fusion protein was detected by RT-PCR and Western blot. Conclusion： The eukaryotic expression vector containing B7-1/GFP gene was constructed successfully, and it could be expressed in LM8 after transfection.
基金a grant from the "95 National Key Project of China" (No.96-906-01-20).
文摘Objective: To study the vaccine potency of gene-modified tumor cells. Methods: The EL-4 lymphoma was transduced with recombinant retrovirus containing the murine GM-CSF gene or B7-1 gene. The effect of gene transduction on antitumor immunity was investigated. Results: Flow cytometry analysis showed that expression of their surface marker between wild-type EL-4 cells and gene transduced tumor cells was the same except for CD80 positive in B7-1 gene transduced cells. GM-CSF gene or B7-1 gene transduced EL-4 cells resulted in remarkable loss of tumorigenicity in syngenetic mice. The systemic protective immunity was induced against the challenge with EL-4/wt cells. Therapeutic vaccine with EL-4/GM-CSF or EL/7-1 cells could retard the growth of established early-stage EL-4/wt tumor significantly, but not retard the growth of late-stage EL-4/wt tumor. Irradiated GM-CSF gene transduced EL-4 cells showed strong vaccine effect against EL-4 cell challenge, but irradiated B7-1 gene transduced EL-4 cells showed weak vaccine effect. Remarkable cooperative antitumor effect against EL-4 cell challenge was observed when both irradiated EL-4/GM-CSF and EL-4/B7-1 were inoculated together. Conclusion: GM-CSF gene or B7-1 gene transduced BL-4 cells can be used as a good tumor vaccine. The combination of the two kinds of vaccine may have potential application value in human cancer treatment.
文摘Objective: To construct recombinant R7-l/B7-2 retrovirus vectors and observe the effects of B7-l/R7-2 gene expression on in ho and in for immune response against against murine hepatoma. Methods: The recombinant retrovirus vectors expressing B7-1/B7-2 were constructed by gene cloning technology to produce retrovirus-infected PE501 and PA317 cell lines and murine hepatoma Hepal-6. The expression of R7-l/B7-2 was detected by fluorescence activated cell soning analysis (FACS). B7-l/B7-2 positive Hepal-6 Cell lines were used in inducing anti-hepatoma immunity in ho and in the. Results: In contrast to the excessive growth of parental Hemal-6 tumor, the growth of B7-l/B7-2-positive Hepal-6 inoculated into syngenic mice regressed. B7-1/R7-2-positive or cytokine-treated Hepal-6 alone could only induce mild cytototicity; in contrast, B7-1/B7-2-positive Hemal-6 treated with cytokine-stimulated spleen cells and activated the cytotoxicity effectively. Immunity in mice with R7-1/B7-2-positive tumor cells or cytokine-beated Hepal-6 only provided partial protection against parental Hepa1-6 tumor, whereas pretreatment of the transfected tumor cells with IFN-r and TNF-a induced complete immunity protection in vivo. Mice receiving inoculation of cytokine-treated B7-l/R7-2-positive Hemal-6 cells presented regression of the establoshed pental tUmor and survived for more than l00 d, while those untreated mice died within 40 d. Conclu sions: B7-l/R7-2 expression is necessary but not sufficient in inducing anti-hepatoma immune response, whereas it is efficient when combined with the beatment of IFN-γ and TNF-a.
文摘A new degraded sesquiterpene was isolated from the marine actinomycete Streptomyces sp. 0616208. Its structure was elucidated as （1α; 4aα; 5α, 7β, 8aβ）-5, 8a-dimethyl-decahydrona- phthalene-1, 4a, 7-triol on the basis of spectroscopic data.
文摘目的 对从混合培养的海洋软珊瑚共附生真菌中获得的结构新颖、活性良好的杂萜类成分进行研究。方法采用大米培养基对2株软珊瑚共附生曲霉属真菌EGF7-0-1和EGF15-0-3进行共培养;根据杂萜类化合物的结构特征,在基于质谱的分子网络(GNPS)和薄层色谱(TLC)的共同指导下,运用多种柱色谱及高效液相色谱(HPLC)等方法对其杂萜类成分进行目标导向分离;通过核磁共振(NMR)、高分辨质谱(HRMS)和旋光(ORD)等技术及物理常数对照等手段对单体化合物进行结构鉴定;采用液相色谱质谱联用(LC-MS^(n))和GNPS技术分析比较获得的杂萜类成分在2株真菌共培养及EGF7-0-1单培养中的含量。结果 从2株海洋真菌Aspergillus sp. EGF7-0-1和EGF15-0-3共培养的大米培养基中共分离得到7个杂萜类化合物,结构分别为terretonin (1)、terretonin A (2)、terretonin D1 (3)、terretonin H (4)、terreustoxin E (5)、aperterpene N (6)和asperterpene J (7)。LC-MSn和GNPS分析结果表明共培养条件下杂萜类化合物更为丰富。结论 化合物1~7均为具6/6/6骈并骨架的3,5-二甲基苔色酸(DMOA)途径衍生而成的杂萜。与菌株EGF7-0-1单培养相比,2株真菌共培养可以诱导产生丰富的杂萜,为进一步新颖杂萜类化合物的挖掘提供支持数据。
