Science in China. Series B, Chemistry, life sciences & earth sciences最新文献_第5页

Construction and high expression of retroviral vector with human clotting factor IX cDNA in vitro. 人凝血因子IX cDNA逆转录病毒载体的构建及体外高表达。

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-06-10 DOI: 10.1360/YB1995-38-6-705

D. Lu, X. Qiu, B. Zheng, J. Xue

{"title":"Construction and high expression of retroviral vector with human clotting factor IX cDNA in vitro.","authors":"D. Lu, X. Qiu, B. Zheng, J. Xue","doi":"10.1360/YB1995-38-6-705","DOIUrl":"https://doi.org/10.1360/YB1995-38-6-705","url":null,"abstract":"The construction of the high titer and highly expressed safety retroviral vector carrying human clotting factor IX cDNA is reported. Retroviral vectors LNCIX, LIXSN and LCIXSN, driven by hCMV, LTR and hCMV combined with LTR promoter respectively, were constructed, based on the retroviral vector LNL6, and transferred into packaging cell line PA317 with electroporation. Human clotting factor IX was detected in the cultured cells transduced with LNCIX and LIXSN but not in the cells transduced with LCIXSN. The viral titer of PA317/LNCIX was 800,000 CFU per mL. With ELISA detection, it was found that the cells transduced with this vector can express human clotting factor IX at the level of 3.3 micrograms per 10(6) cells in 24 h in human fibrosarcoma cells HT-1080 and 2.5 micrograms per 10(6) cells in 24 h in hemophilia B patients' skin fibroblast HSF cells, and more than 80% of them were biologically active. The viral titer and expression of human FIX were increased, and the construction of retroviral vector backbone was improved and the safety was guaranteed as compared to those vectors used previously. These vectors may produce a sufficient quantity of factor IX proteins to cause the phenotypic modification for hemophilia B patients.","PeriodicalId":21648,"journal":{"name":"Science in China. Series B, Chemistry, life sciences & earth sciences","volume":"54 1","pages":"705-12"},"PeriodicalIF":0.0,"publicationDate":"1995-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87901871","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

青藏高原若尔盖盆地RH孔沉积有机质的 δ 13 C值和氢指数记录青藏高原若尔盖盆地RH孔沉积有机质的 δ 13 C值和氢指数记录

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-06-10 DOI: 10.1360/ZB1995-25-6-631

张平中, 王先彬, 陈践发, 李春园, 王苏民

引用次数: 0

Transmembrane Ca2+ gradient-mediated phosphatidylcholine modulating sarcoplasmic reticulum C(a2+)-ATPase. 跨膜Ca2+梯度介导的磷脂酰胆碱调节肌浆网C(a2+)- atp酶。

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-06-10 DOI: 10.1360/YB1995-38-6-713

Y. Tu, Hong Xu, Fu-yu Yang

{"title":"Transmembrane Ca2+ gradient-mediated phosphatidylcholine modulating sarcoplasmic reticulum C(a2+)-ATPase.","authors":"Y. Tu, Hong Xu, Fu-yu Yang","doi":"10.1360/YB1995-38-6-713","DOIUrl":"https://doi.org/10.1360/YB1995-38-6-713","url":null,"abstract":"The sarcoplasmic reticulum (SR) C(a2+)-ATPase was purified and reconstituted into the sealed phospholipids vesicles with or without transmembrane Ca2+ gradient. The role of phospholipids, especially phosphatidylcholine (PC), in the modulation of C(a2+)-ATPase by transmembrane Ca2+ gradient was investigated. The results are as follows. (i) Incubated with phospholipids, the enzyme activity of the delipidated C(a2+)-ATPase is inhibited by Ca2+ and the highest inhibition is observed in the presence of PC. (ii) When there exists a transmembrane Ca2+ gradient (higher Ca2+ concentration inside vesicles, 1,000 mumol/L:50 mumol/L, similar to the physiological condition), the inhibition of C(a2+)-ATPase by transmembrane Ca2+ gradient can be only observed in the vesicles containing PC:PE, but not in those containing PS:PE or PG:PE. The highest inhibition is obtained at a 50:50 molar ratio of PC:PE (iii) By comparing the effects of PC differing in acyl chains, higher inhibition of C(a2+)-ATPase is observed in vesicles containing DPPC:PE and DOPC:PE, while no inhibition in DMPC:PE vesicles (iv) If the transmembrane Ca2+ gradient is in the inverse direction, the enzyme activity of C(a2+)-ATPase is inhibited whenever reconstituted with acidic or neutral phospholipids.","PeriodicalId":21648,"journal":{"name":"Science in China. Series B, Chemistry, life sciences & earth sciences","volume":"42 1","pages":"713-21"},"PeriodicalIF":0.0,"publicationDate":"1995-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85251752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Eu(DBM) 3 4,4′-BIPY的晶体结构和荧光光谱 Eu(DBM) 3 4,4′-BIPY的晶体结构和荧光光谱

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-06-10 DOI: 10.1360/ZB1995-25-6-561

金林培刘世雄蔡冠梁黄金陵秦伟平黄世华王明昭, Jin Lin-Pei Wang Zhe-Min Ca Guan-Liang Zhang Jia-Hua Wang Meng-Zhao

引用次数: 0

Construction and high expression of retroviral vector with human clotting factor IX cDNA in vitro. 人凝血因子IX cDNA逆转录病毒载体的构建及体外高表达。

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-06-01

D Lu, X Qiu, B Zheng, X Qiu, J Xue

{"title":"Construction and high expression of retroviral vector with human clotting factor IX cDNA in vitro.","authors":"D Lu, X Qiu, B Zheng, X Qiu, J Xue","doi":"","DOIUrl":"","url":null,"abstract":"The construction of the high titer and highly expressed safety retroviral vector carrying human clotting factor IX cDNA is reported. Retroviral vectors LNCIX, LIXSN and LCIXSN, driven by hCMV, LTR and hCMV combined with LTR promoter respectively, were constructed, based on the retroviral vector LNL6, and transferred into packaging cell line PA317 with electroporation. Human clotting factor IX was detected in the cultured cells transduced with LNCIX and LIXSN but not in the cells transduced with LCIXSN. The viral titer of PA317/LNCIX was 800,000 CFU per mL. With ELISA detection, it was found that the cells transduced with this vector can express human clotting factor IX at the level of 3.3 micrograms per 10(6) cells in 24 h in human fibrosarcoma cells HT-1080 and 2.5 micrograms per 10(6) cells in 24 h in hemophilia B patients' skin fibroblast HSF cells, and more than 80% of them were biologically active. The viral titer and expression of human FIX were increased, and the construction of retroviral vector backbone was improved and the safety was guaranteed as compared to those vectors used previously. These vectors may produce a sufficient quantity of factor IX proteins to cause the phenotypic modification for hemophilia B patients.","PeriodicalId":21648,"journal":{"name":"Science in China. Series B, Chemistry, life sciences & earth sciences","volume":"38 6","pages":"705-12"},"PeriodicalIF":0.0,"publicationDate":"1995-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18630252","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Transmembrane Ca2+ gradient-mediated phosphatidylcholine modulating sarcoplasmic reticulum C(a2+)-ATPase. 跨膜Ca2+梯度介导的磷脂酰胆碱调节肌浆网C(a2+)- atp酶。

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-06-01

Y Tu, H Xu, F Yang

{"title":"Transmembrane Ca2+ gradient-mediated phosphatidylcholine modulating sarcoplasmic reticulum C(a2+)-ATPase.","authors":"Y Tu, H Xu, F Yang","doi":"","DOIUrl":"","url":null,"abstract":"The sarcoplasmic reticulum (SR) C(a2+)-ATPase was purified and reconstituted into the sealed phospholipids vesicles with or without transmembrane Ca2+ gradient. The role of phospholipids, especially phosphatidylcholine (PC), in the modulation of C(a2+)-ATPase by transmembrane Ca2+ gradient was investigated. The results are as follows. (i) Incubated with phospholipids, the enzyme activity of the delipidated C(a2+)-ATPase is inhibited by Ca2+ and the highest inhibition is observed in the presence of PC. (ii) When there exists a transmembrane Ca2+ gradient (higher Ca2+ concentration inside vesicles, 1,000 mumol/L:50 mumol/L, similar to the physiological condition), the inhibition of C(a2+)-ATPase by transmembrane Ca2+ gradient can be only observed in the vesicles containing PC:PE, but not in those containing PS:PE or PG:PE. The highest inhibition is obtained at a 50:50 molar ratio of PC:PE (iii) By comparing the effects of PC differing in acyl chains, higher inhibition of C(a2+)-ATPase is observed in vesicles containing DPPC:PE and DOPC:PE, while no inhibition in DMPC:PE vesicles (iv) If the transmembrane Ca2+ gradient is in the inverse direction, the enzyme activity of C(a2+)-ATPase is inhibited whenever reconstituted with acidic or neutral phospholipids.","PeriodicalId":21648,"journal":{"name":"Science in China. Series B, Chemistry, life sciences & earth sciences","volume":"38 6","pages":"713-21"},"PeriodicalIF":0.0,"publicationDate":"1995-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18630253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

青藏高原东北部降水中 δ 18 O的变化特征青藏高原东北部降水中 δ 18 O的变化特征

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-05-10 DOI: 10.1360/ZB1995-25-5-540

施雅风姚檀栋章新平, Shi Ya-Feng Tao Shan-Dong Zhang Xin-Beng

引用次数: 0

Abrupt reduction of c-myc expression by antisense RNA inducing terminal differentiation and apoptosis of a human esophageal cancer cell line 通过反义RNA诱导人食管癌细胞系终末分化和凋亡的c-myc表达突然降低

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-05-10 DOI: 10.1360/YB1995-38-5-580

Zhao Xiao-hang, W. Xiuqin, Zhou Chuannong, Peng Renling, Yan Shuizhong, W. Min

{"title":"Abrupt reduction of c-myc expression by antisense RNA inducing terminal differentiation and apoptosis of a human esophageal cancer cell line","authors":"Zhao Xiao-hang, W. Xiuqin, Zhou Chuannong, Peng Renling, Yan Shuizhong, W. Min","doi":"10.1360/YB1995-38-5-580","DOIUrl":"https://doi.org/10.1360/YB1995-38-5-580","url":null,"abstract":"A human esophageal cancer cell line (EC8712) expressing high-level Myc protein was infected with recombmant retroviral particles (pA-BD9) at a multiplicity of infection (MOI) 1:1. This viral particle contains a neomycin-resistant gene and a 1.53-kb antisense RNA spanning the 2nd exon and its flanking sequences of the human c-myc oncogene. The G418-resistant EC8712 clones showed an 86% inhibition of growth rate and morphological changes characteristic of terminal differentiation and apoptosis. A decrease of about 80% of Myc protein was also observed in these infected cells by ABC-ELISA assay. 12-24 h after the infection of ECS712 cells with pA-BD9 at a high viral particle concentration (MOI = 1:10), the integration of the extrinsic 1.53-kb antisense c-myc fragment into the cancer cell genome was evidenced by the Southern blot analysis. Northern blot analyses showed the expression of this antisense fragment and a decrease of the intrinsic c-myc expression by 74% in comparison with that of the parental EC8712 cells. Heterotransplants of the infected EC8712 cells into the nude mice revealed a substantial decrease in tumorigemcity and morphological changes characteristic of terminal differentiation and apoptosis. Primary monolayer cell cultures of normal epithelia derived from the fetal and adult esophagus mucosa were set as controls. No noticeable increase in c-myc expression was found in these cultures Infection of these cells with the same recombinant viral particles neither affected the growth rate of the cells nor their normal morphology. Our experiments indicate that the drastic decrease of the over-expressed Myc protein in cancer cells may also be an entrance to one of many pathways leading to the terminal differentiation and programmed cell death.","PeriodicalId":21648,"journal":{"name":"Science in China. Series B, Chemistry, life sciences & earth sciences","volume":"25 1","pages":"580-589"},"PeriodicalIF":0.0,"publicationDate":"1995-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83404868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 8

Analysis and PCR detection of antigen compositions of ovine progressive pneumonia virus. 绵羊进行性肺炎病毒抗原组成分析及PCR检测。

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-05-10 DOI: 10.1360/YB1995-38-5-573

Ding Enyu

{"title":"Analysis and PCR detection of antigen compositions of ovine progressive pneumonia virus.","authors":"Ding Enyu","doi":"10.1360/YB1995-38-5-573","DOIUrl":"https://doi.org/10.1360/YB1995-38-5-573","url":null,"abstract":"Abstract Ovine progressive pneumonia virus (OPPV) was proliferated utilizing sheep foetal lung cells, and the cytopathic effect (CPE) of the virus was investigated. OPPV was purified with a 10%-sucrose cushion and then with 20%-55% discontinuous sucrose density gradient centrifugation. The structural proteins and antigen compositions of OPPV were analysed by SDS-PAGE and Western blotting. Besides, the OPP proviral cDNAs of the virus-infected cell cultures and the peripheral blood monocytes from sheep infected by the virus were detected using polymerase chain reaction (PCR). The results show that the CPE of sheep foetal lung cells infected by OPPV is typical of the disease. The purified virions are intact and of high purity when observed with an electron microscope. OPPV proteins consist of 18 polypeptide bands and the molecular weights range from 18 to 120 kd. Among these, 3 were glycoproteins (designated by gp120, gp50 and gp47). The appearance and peak time of the p28 antibody from sheep inoculated with OPPV are earlier than those of the p94 antibody to OPPV. Besides, the strength of immune reaction of p28 antibody is greater than that of p94 antibody. With PCR, it is demonstrated that the initial time for OPP proviral cDNAs to be integrated into the sheep foetal lung cells and the peripheral blood monocytes in sheep were 24 h and 9 d after inoculation, respectively.","PeriodicalId":21648,"journal":{"name":"Science in China. Series B, Chemistry, life sciences & earth sciences","volume":"5 1","pages":"573-579"},"PeriodicalIF":0.0,"publicationDate":"1995-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77963101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Interaction between intercalation type anticancer drugs and DNA studied by ultraviolet resonance Raman spectroscopy. 用紫外共振拉曼光谱研究嵌入型抗癌药物与DNA的相互作用。

Science in China. Series B, Chemistry, life sciences & earth sciences Pub Date : 1995-05-10 DOI: 10.1360/YB1995-38-5-555

X. Zhao, D. Lu, S. Jiang, C. Mao, Y. Fan, C. An, Z. Li

引用次数: 5