In the present study, a new member of melanoma associated antigens (Mage), named Restin (219 amino acids), was identified from HL-60 cell induced by all-trans-retinoic acid (ATRA) by PCR-based subtractive hybridizatio...In the present study, a new member of melanoma associated antigens (Mage), named Restin (219 amino acids), was identified from HL-60 cell induced by all-trans-retinoic acid (ATRA) by PCR-based subtractive hybridization. Bioinformatics analysis found this novel gene shares high homolog with Necdin (a neuronal growth suppressor, 49%). Both of them are basic proteins. Moreover, the Restin, Necdin and Mages are in one protein superfamily. This fact indicates that the Restin and Mages are mutually related but functionally different. Further analysis found that they can be divided into two subgroups, the acid and the basic. Restin, Necdin and Mage-D1 have an alkaline conserve region (PI is from 8.6 to 10.1), which are not or less expressed in tumor tissues but mostly in normal tissues. It has been reported that Necdin can arrest the cell proliferation by interaction with p53 and E2F1. Therefore, all of them are probably related to arrest the cell cycle. However, the Mage A and C are primarily acid proteins (PI is from 4.2 to 4.9), not expressed in normal tissues but in tumors. It is quite probable that these proteins are involved in the cell proliferation. We therefore suggest that these two protein families might be a pair of control elements of cell cycle-l'in cycle or out of cycle'.展开更多
Restin,a member of melanoma-associated antigen superfamily gene,was first cloned from differentiated leukemia cell induced by all trans-retinoic acid,and was able to inhibit cell proliferation,but the molecular mechan...Restin,a member of melanoma-associated antigen superfamily gene,was first cloned from differentiated leukemia cell induced by all trans-retinoic acid,and was able to inhibit cell proliferation,but the molecular mechanism was not clear.Since Restin was localized in cell nucleus,and its homolog member,Necdin(neuronal growth suppressor factor),could interact with transcription factors p53 and E2F1,we proposed that Restin might also function as Necdin through interacting with some transcription factors.In this study,transcription factors p53,AP1,ATFs and E2Fs were cloned and used in the mammalian two-hybrid system to identify their in-teraction with Restin.The results showed that only ATF3 had a strong interaction with Restin.It is interesting to know that ATF3 was an important transcription factor for G1 cell cycle initiation in physiological stress response.It was possible that the inhibition of cell proliferation by Restin might be related with the inhibition of ATF3 activity.展开更多
Restin, belonging to the melanoma-associated antigen superfamily, was firstly cloned from the differentiated HL-60 cells when induced by all-trans retinoic acid ( ATRA ) in our lab. Our previous results showed that re...Restin, belonging to the melanoma-associated antigen superfamily, was firstly cloned from the differentiated HL-60 cells when induced by all-trans retinoic acid ( ATRA ) in our lab. Our previous results showed that restin might be correlated to cell cycle arrest. Due to the importance of p53 in the regulation of cell growth and the relationship between p53 and ATRA, we tried to test the relationship between p53 and restin. Firstly, transfection results showed that p53 was able to upregulate the expression of restin at the transcriptional level when p53 was transfected into eukaryotic cells. Secondly, the bioinformatics analysis revealed that the upstream sequence (about 2 kb) from the first ATG of the ORF of restin gene contained a p53 binding site. In order to confirm that p53 was involved in the transcriptional regulation of restin, we cloned the upstream sequence of restin and constructed the promoter luciferase reporter system. From the luciferase activity, we demonstrated that the promoter of restin gene could be induced by ATRA. Then, another two luciferase reporter plasmids driven by the reporter of restin with no (RP?p53-luc) or mutant (mRP-luc) p53 binding site were constructed to see the regulation of restin by p53. Results showed that the transcriptional upregulation of restin gene was not due to the putative p53 binding site on the upstream of restin gene. We proposed that p53 upregulated restin transcription through an indirect way rather than direct interaction with the cis-activating element of the restin promoter.展开更多
目的:优选大孔吸附树脂纯化沙苑子总黄酮的工艺。方法:考察AB-8、D-101和NKA-9树脂对沙苑子总黄酮的吸附及解吸性能,筛选树脂型号;以总黄酮含量为指标,采用单因素试验和正交试验优选AB-8树脂纯化工艺条件。结果:选用AB-8型大孔吸附树脂...目的:优选大孔吸附树脂纯化沙苑子总黄酮的工艺。方法:考察AB-8、D-101和NKA-9树脂对沙苑子总黄酮的吸附及解吸性能,筛选树脂型号;以总黄酮含量为指标,采用单因素试验和正交试验优选AB-8树脂纯化工艺条件。结果:选用AB-8型大孔吸附树脂,优选的工艺条件为:上样液质量浓度10 mg·m L-1,样品液p H 5.0,上样速度为1.0 m L·min-1,乙醇洗脱浓度40%,洗脱流速为1.5 m L·min-1;经AB-8处理后的沙苑子总黄酮纯度达50%。结论:AB-8型大孔吸附树脂能有效地纯化富集沙苑子总黄酮。展开更多
Wang Jiyong, among the thousands of retail sellers who keep Beijing's inhabitants and restaurants supplied with fresh vegetables, left his little village in Henan Province 17 years ago, later to be joined by his w...Wang Jiyong, among the thousands of retail sellers who keep Beijing's inhabitants and restaurants supplied with fresh vegetables, left his little village in Henan Province 17 years ago, later to be joined by his wife, Zhang Chunling, his brother, his son and his father. With perseverance and determination, and a well-organized distribution of work within the family, they manage to make a living not only for themselves, but also for members of the family who live back in their home town.展开更多
《英语世界》94年第一期16页第31段(英汉对照读物)有这样一段话: 原文:Once,about 20 feet away,a bear rose on its haunches to stare downat her.Don’t move!She thought,Don’t even blink! 译文:有一次,离她二十英尺的地方,一...《英语世界》94年第一期16页第31段(英汉对照读物)有这样一段话: 原文:Once,about 20 feet away,a bear rose on its haunches to stare downat her.Don’t move!She thought,Don’t even blink! 译文:有一次,离她二十英尺的地方,一只熊蹲在后腿上,直着上身,一双眼睛直盯着她。“不能动!”她想到,“甚至眼睛都不能眨!”展开更多
文摘In the present study, a new member of melanoma associated antigens (Mage), named Restin (219 amino acids), was identified from HL-60 cell induced by all-trans-retinoic acid (ATRA) by PCR-based subtractive hybridization. Bioinformatics analysis found this novel gene shares high homolog with Necdin (a neuronal growth suppressor, 49%). Both of them are basic proteins. Moreover, the Restin, Necdin and Mages are in one protein superfamily. This fact indicates that the Restin and Mages are mutually related but functionally different. Further analysis found that they can be divided into two subgroups, the acid and the basic. Restin, Necdin and Mage-D1 have an alkaline conserve region (PI is from 8.6 to 10.1), which are not or less expressed in tumor tissues but mostly in normal tissues. It has been reported that Necdin can arrest the cell proliferation by interaction with p53 and E2F1. Therefore, all of them are probably related to arrest the cell cycle. However, the Mage A and C are primarily acid proteins (PI is from 4.2 to 4.9), not expressed in normal tissues but in tumors. It is quite probable that these proteins are involved in the cell proliferation. We therefore suggest that these two protein families might be a pair of control elements of cell cycle-l'in cycle or out of cycle'.
文摘Restin,a member of melanoma-associated antigen superfamily gene,was first cloned from differentiated leukemia cell induced by all trans-retinoic acid,and was able to inhibit cell proliferation,but the molecular mechanism was not clear.Since Restin was localized in cell nucleus,and its homolog member,Necdin(neuronal growth suppressor factor),could interact with transcription factors p53 and E2F1,we proposed that Restin might also function as Necdin through interacting with some transcription factors.In this study,transcription factors p53,AP1,ATFs and E2Fs were cloned and used in the mammalian two-hybrid system to identify their in-teraction with Restin.The results showed that only ATF3 had a strong interaction with Restin.It is interesting to know that ATF3 was an important transcription factor for G1 cell cycle initiation in physiological stress response.It was possible that the inhibition of cell proliferation by Restin might be related with the inhibition of ATF3 activity.
基金the National Natural Science Foundation of China (Grant Nos. 30271457 and 30470874)
文摘Restin, belonging to the melanoma-associated antigen superfamily, was firstly cloned from the differentiated HL-60 cells when induced by all-trans retinoic acid ( ATRA ) in our lab. Our previous results showed that restin might be correlated to cell cycle arrest. Due to the importance of p53 in the regulation of cell growth and the relationship between p53 and ATRA, we tried to test the relationship between p53 and restin. Firstly, transfection results showed that p53 was able to upregulate the expression of restin at the transcriptional level when p53 was transfected into eukaryotic cells. Secondly, the bioinformatics analysis revealed that the upstream sequence (about 2 kb) from the first ATG of the ORF of restin gene contained a p53 binding site. In order to confirm that p53 was involved in the transcriptional regulation of restin, we cloned the upstream sequence of restin and constructed the promoter luciferase reporter system. From the luciferase activity, we demonstrated that the promoter of restin gene could be induced by ATRA. Then, another two luciferase reporter plasmids driven by the reporter of restin with no (RP?p53-luc) or mutant (mRP-luc) p53 binding site were constructed to see the regulation of restin by p53. Results showed that the transcriptional upregulation of restin gene was not due to the putative p53 binding site on the upstream of restin gene. We proposed that p53 upregulated restin transcription through an indirect way rather than direct interaction with the cis-activating element of the restin promoter.
文摘目的:优选大孔吸附树脂纯化沙苑子总黄酮的工艺。方法:考察AB-8、D-101和NKA-9树脂对沙苑子总黄酮的吸附及解吸性能,筛选树脂型号;以总黄酮含量为指标,采用单因素试验和正交试验优选AB-8树脂纯化工艺条件。结果:选用AB-8型大孔吸附树脂,优选的工艺条件为:上样液质量浓度10 mg·m L-1,样品液p H 5.0,上样速度为1.0 m L·min-1,乙醇洗脱浓度40%,洗脱流速为1.5 m L·min-1;经AB-8处理后的沙苑子总黄酮纯度达50%。结论:AB-8型大孔吸附树脂能有效地纯化富集沙苑子总黄酮。
文摘Wang Jiyong, among the thousands of retail sellers who keep Beijing's inhabitants and restaurants supplied with fresh vegetables, left his little village in Henan Province 17 years ago, later to be joined by his wife, Zhang Chunling, his brother, his son and his father. With perseverance and determination, and a well-organized distribution of work within the family, they manage to make a living not only for themselves, but also for members of the family who live back in their home town.
文摘《英语世界》94年第一期16页第31段(英汉对照读物)有这样一段话: 原文:Once,about 20 feet away,a bear rose on its haunches to stare downat her.Don’t move!She thought,Don’t even blink! 译文:有一次,离她二十英尺的地方,一只熊蹲在后腿上,直着上身,一双眼睛直盯着她。“不能动!”她想到,“甚至眼睛都不能眨!”