目的探讨基于SNaPshot技术的精液特异性编码区单核苷酸多态性(coding region single nucleo⁃tide polymorphism,cSNP)遗传标记检测在精液(斑)溯源及混合体液(斑)鉴定中的可行性。方法制备16例精液斑和11例精液-静脉血混合斑样本,提取其...目的探讨基于SNaPshot技术的精液特异性编码区单核苷酸多态性(coding region single nucleo⁃tide polymorphism,cSNP)遗传标记检测在精液(斑)溯源及混合体液(斑)鉴定中的可行性。方法制备16例精液斑和11例精液-静脉血混合斑样本,提取其基因组DNA(genomic DNA,gDNA)和总RNA,并将总RNA逆转录为互补DNA(complementary DNA,cDNA)。在已验证的精液特异性mRNA编码基因上筛选cSNP遗传标记。基于SNaPshot技术构建cSNP复合检测体系并通过CE对样本进行基因分型。结果成功构建了包含5个精液特异性cSNP的复合检测体系。在16例精液样本中,除了位于TGM4基因上的cSNP在cDNA的检测结果中出现等位基因丢失外,其余cSNP的gDNA和cDNA分型结果高度一致。检测精液-静脉血混合斑时,检出的cSNP分型结果均与精液提供者的基因型一致,未受到静脉血提供者基因型的干扰。结论应用SNaPshot技术检测精液特异性cSNP的方法可以应用于法医学精液(斑)的基因分型,并为混合体液(斑)中精液来源个体的判定提供信息。展开更多
To improve the accuracy and reduce the calcu- lation cost for the inverse problem of centrifugal pump impeller, the new inverse method based on proper orthog- onal decomposition （POD） is proposed. The pump blade sha...To improve the accuracy and reduce the calcu- lation cost for the inverse problem of centrifugal pump impeller, the new inverse method based on proper orthog- onal decomposition （POD） is proposed. The pump blade shape is parameterized by quartic Bezier curve, and the initial snapshots is generated by introducing the perturbation of the blade shape control parameters. The internal flow field and its hydraulic performance is predicted by CFD method. The snapshots vector includes the blade shape parameter and the distribution of blade load. The POD basis for the snap- shots set are deduced by proper orthogonal decomposition. The sample vector set is expressed in terms of the linear combination of the orthogonal basis. The objective blade shape corresponding to the objective distribution of blade load is obtained by least square fit. The Iterative correction algorithm for the centrifugal pump blade inverse method based on POD is proposed. The objective blade load dis- tributions are corrected according to the difference of the CFD result and the POD result. The two dimensional and three dimensional blade calculation cases show that the proposed centrifugal pump blade inverse method based on POD have good convergence and high accuracy, and thecalculation cost is greatly reduced. After two iterations, the deviation of the blade load and the pump hydraulic perfor- mance are limited within 4.0% and 6.0% individually for most of the flow rate range. This paper provides a promising inverse method for centrifugal pump impeller, which will benefit the hydraulic optimization of centrifugal pump.展开更多
The introduction of the multi well plate miniaturisation technology with its associated automated dispensers, readers and integrated systems coupled with advances in life sciences has a propelling effect on the rate ...The introduction of the multi well plate miniaturisation technology with its associated automated dispensers, readers and integrated systems coupled with advances in life sciences has a propelling effect on the rate at which new potential drug molecules are discovered. The translation of these discoveries to real outcome now demands parallel approaches which allow large numbers of process options to be rapidly assessed. The engineering challenges in achieving this provide the motivation for the proposed work. In this work we used computational fluid dynamics(CFD) analysis to study flow conditions in a gas liquid contactor which has the potential to be used as a fermenter on a multi well format. The bioreactor had a working volume of 6 5 mL with the major dimensions equal to those of a single well of a 24 well plate. The 6 5 mL bioreactor was mechanically agitated and aerated by a single sparger placed beneath the bottom impeller. Detailed numerical procedure for solving the governing flow equations is given. The CFD results are combined with population balance equations to establish the size of the bubbles and their distribution in the bioreactor. Power curves with and without aeration are provided based on the simulated results.展开更多
目的使用R6G-ddATP作为双脱氧荧光底物建立单碱基末端延伸(SNaPShot)-凝胶荧光法快速检测3种高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)(HPV18、HPV33、HPV35)基因型。方法使用HPV质控品作为样本,R6G-ddATP双脱氧荧光...目的使用R6G-ddATP作为双脱氧荧光底物建立单碱基末端延伸(SNaPShot)-凝胶荧光法快速检测3种高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)(HPV18、HPV33、HPV35)基因型。方法使用HPV质控品作为样本,R6G-ddATP双脱氧荧光试剂作为底物,首先利用通用引物对HPV进行扩增,得到第一轮扩增产物,经纯化后作为后续SNaPShot反应的模板;然后利用特异性的一步延伸引物进行SNaPShot反应,生成带有R6G荧光标记的DNA延伸产物;产物经过琼脂糖凝胶电泳,在凝胶成像仪下观察电泳结果,通过不同的一步延伸引物对HPV进行分型。每个样本均重复检测3次,并与DNA测序结果进行比较。结果优选的SNaPShot反应的退火温度为55℃;仅需3 h即可对HPV进行分型;在该最适条件下使用R6G-ddATP/SNaPShot-凝胶荧光法检测3种HPV基因型,检测结果与测序结果一致。结论成功建立了3种HR-HPV基因型的微量检测方法——R6G-ddATP/SNaPShot-凝胶荧光法,可用于HPV基因型的快速检测。展开更多
文摘目的探讨基于SNaPshot技术的精液特异性编码区单核苷酸多态性(coding region single nucleo⁃tide polymorphism,cSNP)遗传标记检测在精液(斑)溯源及混合体液(斑)鉴定中的可行性。方法制备16例精液斑和11例精液-静脉血混合斑样本,提取其基因组DNA(genomic DNA,gDNA)和总RNA,并将总RNA逆转录为互补DNA(complementary DNA,cDNA)。在已验证的精液特异性mRNA编码基因上筛选cSNP遗传标记。基于SNaPshot技术构建cSNP复合检测体系并通过CE对样本进行基因分型。结果成功构建了包含5个精液特异性cSNP的复合检测体系。在16例精液样本中,除了位于TGM4基因上的cSNP在cDNA的检测结果中出现等位基因丢失外,其余cSNP的gDNA和cDNA分型结果高度一致。检测精液-静脉血混合斑时,检出的cSNP分型结果均与精液提供者的基因型一致,未受到静脉血提供者基因型的干扰。结论应用SNaPshot技术检测精液特异性cSNP的方法可以应用于法医学精液(斑)的基因分型,并为混合体液(斑)中精液来源个体的判定提供信息。
基金Supported by National Natural Science Foundation of China(Grant Nos.51469014,51676003)National Key Research and Development Program of China(Grant No.20016YFB0200901)
文摘To improve the accuracy and reduce the calcu- lation cost for the inverse problem of centrifugal pump impeller, the new inverse method based on proper orthog- onal decomposition （POD） is proposed. The pump blade shape is parameterized by quartic Bezier curve, and the initial snapshots is generated by introducing the perturbation of the blade shape control parameters. The internal flow field and its hydraulic performance is predicted by CFD method. The snapshots vector includes the blade shape parameter and the distribution of blade load. The POD basis for the snap- shots set are deduced by proper orthogonal decomposition. The sample vector set is expressed in terms of the linear combination of the orthogonal basis. The objective blade shape corresponding to the objective distribution of blade load is obtained by least square fit. The Iterative correction algorithm for the centrifugal pump blade inverse method based on POD is proposed. The objective blade load dis- tributions are corrected according to the difference of the CFD result and the POD result. The two dimensional and three dimensional blade calculation cases show that the proposed centrifugal pump blade inverse method based on POD have good convergence and high accuracy, and thecalculation cost is greatly reduced. After two iterations, the deviation of the blade load and the pump hydraulic perfor- mance are limited within 4.0% and 6.0% individually for most of the flow rate range. This paper provides a promising inverse method for centrifugal pump impeller, which will benefit the hydraulic optimization of centrifugal pump.
文摘The introduction of the multi well plate miniaturisation technology with its associated automated dispensers, readers and integrated systems coupled with advances in life sciences has a propelling effect on the rate at which new potential drug molecules are discovered. The translation of these discoveries to real outcome now demands parallel approaches which allow large numbers of process options to be rapidly assessed. The engineering challenges in achieving this provide the motivation for the proposed work. In this work we used computational fluid dynamics(CFD) analysis to study flow conditions in a gas liquid contactor which has the potential to be used as a fermenter on a multi well format. The bioreactor had a working volume of 6 5 mL with the major dimensions equal to those of a single well of a 24 well plate. The 6 5 mL bioreactor was mechanically agitated and aerated by a single sparger placed beneath the bottom impeller. Detailed numerical procedure for solving the governing flow equations is given. The CFD results are combined with population balance equations to establish the size of the bubbles and their distribution in the bioreactor. Power curves with and without aeration are provided based on the simulated results.
文摘目的使用R6G-ddATP作为双脱氧荧光底物建立单碱基末端延伸(SNaPShot)-凝胶荧光法快速检测3种高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)(HPV18、HPV33、HPV35)基因型。方法使用HPV质控品作为样本,R6G-ddATP双脱氧荧光试剂作为底物,首先利用通用引物对HPV进行扩增,得到第一轮扩增产物,经纯化后作为后续SNaPShot反应的模板;然后利用特异性的一步延伸引物进行SNaPShot反应,生成带有R6G荧光标记的DNA延伸产物;产物经过琼脂糖凝胶电泳,在凝胶成像仪下观察电泳结果,通过不同的一步延伸引物对HPV进行分型。每个样本均重复检测3次,并与DNA测序结果进行比较。结果优选的SNaPShot反应的退火温度为55℃;仅需3 h即可对HPV进行分型;在该最适条件下使用R6G-ddATP/SNaPShot-凝胶荧光法检测3种HPV基因型,检测结果与测序结果一致。结论成功建立了3种HR-HPV基因型的微量检测方法——R6G-ddATP/SNaPShot-凝胶荧光法,可用于HPV基因型的快速检测。
