AIM: To identify the new alternative splicing variants of human CYP2D6 in human extratumoral liver tissue with RT-PCR and sequencing. METHODS: Full length of human CYP2D6 cDNAs was amplificated by reverse transcriptio...AIM: To identify the new alternative splicing variants of human CYP2D6 in human extratumoral liver tissue with RT-PCR and sequencing. METHODS: Full length of human CYP2D6 cDNAs was amplificated by reverse transcription-polymerase chain reaction (RT-PCR) from a human extratumoral liver tissue and cloned into pGEM-T vector. The cDNA was sequenced. Exons from 1 to 4 of human CYP2D6 cDNAs were also amplificated by RT-PCR from extratumoral liver tissues of 17 human hepatocellular carcinomas. Some RT-PCR products were sequenced. Exons 1 to 4 of CYP2D6 gene were amplified by PCR from extratumoral liver tissue DNA. Two PCR products from extratumoral liver tissues expressing skipped mRNA were partially sequenced. RESULTS: One of the CYP2D6 cDNAs had 470 nucleotides from 79 to 548 (3' portion of exons 1 to 5' portion of exon 4), and was skipped. Exons 1 to 4 of CYP2D6 cDNA were assayed with RT-PCR in 17 extratumoral liver tissues. Both wild type and skipped mRNAs were expressed in 4 samples, only wild type mRNA was expressed in 5 samples, and only skipped mRNA was expressed in 8 samples. Two more variants were identified by sequencing the RT-PCR products of exons 1 to 4 of CYP2D6 cDNA. The second variant skipped 411 nucleotides from 175 to 585. This variant was identified in 4 different liver tissues by sequencing the RT-PCR products. We sequenced partially 2 of the PCR products amplified of CYP2D6 exon 1 to exon 4 from extratumoral liver tissue genomic DNA that only expressed skipped mRNA by RT-PCR. No point mutations around exon 1, intron 1, and exon 4, and no deletion in CYP2D6 gene were detected. The third variant was the skipped exon 3, and 153 bp was lost. CONCLUSION: Three new alternative splicing variants of CYP2D6 mRNA have been identified. They may not be caused by gene mutation and may lose CYP2D6 activity and act as a down-regulator of CYP2D6.展开更多
In this study dynamic analysis of Soil Structure Interaction (SSI) effect on multi story reinforced concrete (RC) frame founded on soft soil (flexible base) is made and compared with fixed base. Two model 2D RC frames...In this study dynamic analysis of Soil Structure Interaction (SSI) effect on multi story reinforced concrete (RC) frame founded on soft soil (flexible base) is made and compared with fixed base. Two model 2D RC frames with 7 and 12 story are selected for analysis. Winkler Spring and half space direct method models are used for flexible base for the frames founded on two types of soft soils with shear velocity Vs < 150 m/s Asper Seismic Codes of Chinese GB50011-2010 Soil IV and Ethiopian ES8-2015 soil D. The frames are subjected to strong ground motion matched to response spectrums of soft soil of Chinese GB50011-2010 and Ethiopian ES8-2015 for linear time history analysis. The dynamic analysis result shows Spring and Fixed base mass participation 90% reaches in 2 or 3 modes but in direct method 11 to 30 modes for story 12 and 7 respectively. However, both flexible base models have bigger fundamental period of vibration and inter story drift but smaller base shear than fixed base. In addition, within the flexible base models the inter-story drift, second order effect (P-Δ) and Story shear distribution are different along the height of frames. The spring model shows larger Story drift and second order effect (P-Δ) at the bottom of Story for both soft soils types. On the other hand, half space direct method model indicates value reverse to spring model;it gives bigger Story drift and P-Δ effect in the top stories than fixed base. Finally, this study concludes that base shear reduction due to SSI may not be always beneficial. Because the gravity load is constant in both fixed and flexible bases that cause bigger P-Δ effect at the bottom stories due to increase, inter story drift and decrease story shear in flexible base.展开更多
基金Supported by the National Key Basic Research and Development Program of China,No.2002CB512901National Natural Science Foundation of China,No.39770868 and Natural Science Foundation of Zhejiang Province,No.397490
文摘AIM: To identify the new alternative splicing variants of human CYP2D6 in human extratumoral liver tissue with RT-PCR and sequencing. METHODS: Full length of human CYP2D6 cDNAs was amplificated by reverse transcription-polymerase chain reaction (RT-PCR) from a human extratumoral liver tissue and cloned into pGEM-T vector. The cDNA was sequenced. Exons from 1 to 4 of human CYP2D6 cDNAs were also amplificated by RT-PCR from extratumoral liver tissues of 17 human hepatocellular carcinomas. Some RT-PCR products were sequenced. Exons 1 to 4 of CYP2D6 gene were amplified by PCR from extratumoral liver tissue DNA. Two PCR products from extratumoral liver tissues expressing skipped mRNA were partially sequenced. RESULTS: One of the CYP2D6 cDNAs had 470 nucleotides from 79 to 548 (3' portion of exons 1 to 5' portion of exon 4), and was skipped. Exons 1 to 4 of CYP2D6 cDNA were assayed with RT-PCR in 17 extratumoral liver tissues. Both wild type and skipped mRNAs were expressed in 4 samples, only wild type mRNA was expressed in 5 samples, and only skipped mRNA was expressed in 8 samples. Two more variants were identified by sequencing the RT-PCR products of exons 1 to 4 of CYP2D6 cDNA. The second variant skipped 411 nucleotides from 175 to 585. This variant was identified in 4 different liver tissues by sequencing the RT-PCR products. We sequenced partially 2 of the PCR products amplified of CYP2D6 exon 1 to exon 4 from extratumoral liver tissue genomic DNA that only expressed skipped mRNA by RT-PCR. No point mutations around exon 1, intron 1, and exon 4, and no deletion in CYP2D6 gene were detected. The third variant was the skipped exon 3, and 153 bp was lost. CONCLUSION: Three new alternative splicing variants of CYP2D6 mRNA have been identified. They may not be caused by gene mutation and may lose CYP2D6 activity and act as a down-regulator of CYP2D6.
文摘In this study dynamic analysis of Soil Structure Interaction (SSI) effect on multi story reinforced concrete (RC) frame founded on soft soil (flexible base) is made and compared with fixed base. Two model 2D RC frames with 7 and 12 story are selected for analysis. Winkler Spring and half space direct method models are used for flexible base for the frames founded on two types of soft soils with shear velocity Vs < 150 m/s Asper Seismic Codes of Chinese GB50011-2010 Soil IV and Ethiopian ES8-2015 soil D. The frames are subjected to strong ground motion matched to response spectrums of soft soil of Chinese GB50011-2010 and Ethiopian ES8-2015 for linear time history analysis. The dynamic analysis result shows Spring and Fixed base mass participation 90% reaches in 2 or 3 modes but in direct method 11 to 30 modes for story 12 and 7 respectively. However, both flexible base models have bigger fundamental period of vibration and inter story drift but smaller base shear than fixed base. In addition, within the flexible base models the inter-story drift, second order effect (P-Δ) and Story shear distribution are different along the height of frames. The spring model shows larger Story drift and second order effect (P-Δ) at the bottom of Story for both soft soils types. On the other hand, half space direct method model indicates value reverse to spring model;it gives bigger Story drift and P-Δ effect in the top stories than fixed base. Finally, this study concludes that base shear reduction due to SSI may not be always beneficial. Because the gravity load is constant in both fixed and flexible bases that cause bigger P-Δ effect at the bottom stories due to increase, inter story drift and decrease story shear in flexible base.
