Objective: To scan single nucleotide polymorphism ( SNP ) in Chinese alpha-2Aadrenergic receptor (α_(2A)-AR) gene and study the effects of the SNP on the gene expression.Methods: The complete sequence of α_(2A)-AR g...Objective: To scan single nucleotide polymorphism ( SNP ) in Chinese alpha-2Aadrenergic receptor (α_(2A)-AR) gene and study the effects of the SNP on the gene expression.Methods: The complete sequence of α_(2A)-AR gene was analyzed with automated DNA sequencer to scanSNPs. Genomic DNA was extracted from whole blood and a 239 bp fragment containing the G/Cpolymorphism was amplified with PCR using a pair of. specific primers. PCR-RFLP was used to performthe genotyping of the SNP at the site-1 296 bp of the people in the North of China. Electrophoresismobility shift assay ( EMSA ) was used to study the binding of the 390 bp fragments (- 1 414-1 025bp) with G or C at the site-1 296 bp and nuclear extracts . Results: In our study, two SNPs werefound in α_(2A)-AR gene. Allele frequencies of the SNP at the site-1 296 bp were 0.61 and 0.39 forG and C , and the genotype frequencies were 0.34 , 0.54 and 0.13 for GG, GC and CC respectively fromthe people in the North of China. In the EMSA, a specific binding appeared in the complex ofnuclear extracts and DNA with C at-1 296 bp . Conclusion: Two SNPs exist in α_(2A)-AR gene from thepeople in the North of China , and DNA fragment with allele C of the SNP at the site-1 296 bp couldbind with a specific protein, which could influence the gene expression.展开更多
A cDNA molecule encoding a major part of the hu-man Norepinephrine transporter(hNET) was synthesized by means of Polymerase Chain Reaction(PCR) technique and used as a probe for selecting the human genomic NET gene. A...A cDNA molecule encoding a major part of the hu-man Norepinephrine transporter(hNET) was synthesized by means of Polymerase Chain Reaction(PCR) technique and used as a probe for selecting the human genomic NET gene. A positive clone harbouring the whole gene was ob-tained from a human lymphocyte genomic library through utilizing the "genomic walking" technique. The clone, des-ignated as phNET, harbours a DNA fragment of about 59 kb in length inserted into BamH Ⅰ site in cosmid pWE15.The genomic clone contains 14 exons encoding all amino acid residues in the protein. A single exon encodes a dis-tinct transmembrane domaill, except for transmembrane domain 10 and 11, which are encoded by part of two ex-ons respectively, and exon 12, which encodes part of do-main 11 and all of domain 12. These results imply that there is a close relationship between exon splicing of a gene and structural domains of the protein, as is the case for the human γ-aminobutyric acid transporter(hGAT) and a number of other membrane proteins.展开更多
The human norepinephrine transporter(NET) gene was cloned and structurally analyzed. The far 5’ fragment containing exon 1 (a non-coding exon) and exon 2 was sequenced. The transcription start site of the gene in hum...The human norepinephrine transporter(NET) gene was cloned and structurally analyzed. The far 5’ fragment containing exon 1 (a non-coding exon) and exon 2 was sequenced. The transcription start site of the gene in human brain stem tissue was determined by primer extension analysis. It was found that the gene could be transcribed from multiple starting points. The 5’ flanking sequence contains a proximal G-C rich region, one possible GSG elemeflt and several SP1 sites. However it does not contain TATA box and CAAT box motifS. Gel shift analysis with nuclear extracts from different tissues of mouse shows that the G-C rich region may be involved in tissue specific expression of the gene.展开更多
The clinical observation revealed that the asthmatic symptoms in most of the patients began to be improved after several acupuncture treatments with the dosage of the drug gradually reduced. Generally, the dose of cor...The clinical observation revealed that the asthmatic symptoms in most of the patients began to be improved after several acupuncture treatments with the dosage of the drug gradually reduced. Generally, the dose of cortisone per os was decreased by 2 mg every 10 days, while that of aerosol was controlled by the patients themselves according to the condition of the disease. The symptoms in most of the patients were markedly improved after 15 treatments, but the treatment should be continued for another 10 times to consolidate the curative effect. Thus, each asthmatic patient needs to receive approximately 30 sessions of acupuncture treatment, lasting about 3 months. Thereafter, in order to prevent its relapse, the treatment should be administered 10 times each year in the summer season. It is worthy to be mentioned that, in this series, there was a female patient who suffered from anaphylactic asthma induced by dog's hair. She received 10 sessions of acupuncture treatment with no any improvement. In this case, the acupuncture treatment should not be given any longer. There were another two children patients aged 5 and 7 years respectively, for them the above method of acupuncture treatment was difficult to be used. Therefore, the auricular pressing method combined with cupping was adopted instead. The auricular points selected were Shenmen, Lung, Large Intestine, sensitive point, Pingchuan, Subcortex, and Kidney. 5 points were used each time, and pressed with the seeds of Vaccaria which were changed every 3 days. The patients were asked to press each of the points themselves 3 times a day (each time by 20 pressings). And Dazhui (Du 14), Feishu (UB 13), Pishu (UB 20) and Shenshu (UB 23) were cupped for 5 min. each time. The auricular pressing and cupping therapies may also be added for adult patients to strengthen the curative effect. The 25 cases of hormone dependent bronchial asthma were treated by acupuncture, yielding a markedly effective rate of 56% with a total effective rate of 96%, indicating that good therapeutic results can also be expected in the western countries for those cases who have already been treated with hormones.展开更多
文摘Objective: To scan single nucleotide polymorphism ( SNP ) in Chinese alpha-2Aadrenergic receptor (α_(2A)-AR) gene and study the effects of the SNP on the gene expression.Methods: The complete sequence of α_(2A)-AR gene was analyzed with automated DNA sequencer to scanSNPs. Genomic DNA was extracted from whole blood and a 239 bp fragment containing the G/Cpolymorphism was amplified with PCR using a pair of. specific primers. PCR-RFLP was used to performthe genotyping of the SNP at the site-1 296 bp of the people in the North of China. Electrophoresismobility shift assay ( EMSA ) was used to study the binding of the 390 bp fragments (- 1 414-1 025bp) with G or C at the site-1 296 bp and nuclear extracts . Results: In our study, two SNPs werefound in α_(2A)-AR gene. Allele frequencies of the SNP at the site-1 296 bp were 0.61 and 0.39 forG and C , and the genotype frequencies were 0.34 , 0.54 and 0.13 for GG, GC and CC respectively fromthe people in the North of China. In the EMSA, a specific binding appeared in the complex ofnuclear extracts and DNA with C at-1 296 bp . Conclusion: Two SNPs exist in α_(2A)-AR gene from thepeople in the North of China , and DNA fragment with allele C of the SNP at the site-1 296 bp couldbind with a specific protein, which could influence the gene expression.
文摘A cDNA molecule encoding a major part of the hu-man Norepinephrine transporter(hNET) was synthesized by means of Polymerase Chain Reaction(PCR) technique and used as a probe for selecting the human genomic NET gene. A positive clone harbouring the whole gene was ob-tained from a human lymphocyte genomic library through utilizing the "genomic walking" technique. The clone, des-ignated as phNET, harbours a DNA fragment of about 59 kb in length inserted into BamH Ⅰ site in cosmid pWE15.The genomic clone contains 14 exons encoding all amino acid residues in the protein. A single exon encodes a dis-tinct transmembrane domaill, except for transmembrane domain 10 and 11, which are encoded by part of two ex-ons respectively, and exon 12, which encodes part of do-main 11 and all of domain 12. These results imply that there is a close relationship between exon splicing of a gene and structural domains of the protein, as is the case for the human γ-aminobutyric acid transporter(hGAT) and a number of other membrane proteins.
文摘The human norepinephrine transporter(NET) gene was cloned and structurally analyzed. The far 5’ fragment containing exon 1 (a non-coding exon) and exon 2 was sequenced. The transcription start site of the gene in human brain stem tissue was determined by primer extension analysis. It was found that the gene could be transcribed from multiple starting points. The 5’ flanking sequence contains a proximal G-C rich region, one possible GSG elemeflt and several SP1 sites. However it does not contain TATA box and CAAT box motifS. Gel shift analysis with nuclear extracts from different tissues of mouse shows that the G-C rich region may be involved in tissue specific expression of the gene.
文摘The clinical observation revealed that the asthmatic symptoms in most of the patients began to be improved after several acupuncture treatments with the dosage of the drug gradually reduced. Generally, the dose of cortisone per os was decreased by 2 mg every 10 days, while that of aerosol was controlled by the patients themselves according to the condition of the disease. The symptoms in most of the patients were markedly improved after 15 treatments, but the treatment should be continued for another 10 times to consolidate the curative effect. Thus, each asthmatic patient needs to receive approximately 30 sessions of acupuncture treatment, lasting about 3 months. Thereafter, in order to prevent its relapse, the treatment should be administered 10 times each year in the summer season. It is worthy to be mentioned that, in this series, there was a female patient who suffered from anaphylactic asthma induced by dog's hair. She received 10 sessions of acupuncture treatment with no any improvement. In this case, the acupuncture treatment should not be given any longer. There were another two children patients aged 5 and 7 years respectively, for them the above method of acupuncture treatment was difficult to be used. Therefore, the auricular pressing method combined with cupping was adopted instead. The auricular points selected were Shenmen, Lung, Large Intestine, sensitive point, Pingchuan, Subcortex, and Kidney. 5 points were used each time, and pressed with the seeds of Vaccaria which were changed every 3 days. The patients were asked to press each of the points themselves 3 times a day (each time by 20 pressings). And Dazhui (Du 14), Feishu (UB 13), Pishu (UB 20) and Shenshu (UB 23) were cupped for 5 min. each time. The auricular pressing and cupping therapies may also be added for adult patients to strengthen the curative effect. The 25 cases of hormone dependent bronchial asthma were treated by acupuncture, yielding a markedly effective rate of 56% with a total effective rate of 96%, indicating that good therapeutic results can also be expected in the western countries for those cases who have already been treated with hormones.
