Selection of proper reference genes (RGs) is an essential step needed for accurate normalization of results from genomic studies. Expression of RGs is regulated by many factors such as species, age, gender, type of ti...Selection of proper reference genes (RGs) is an essential step needed for accurate normalization of results from genomic studies. Expression of RGs is regulated by many factors such as species, age, gender, type of tissue, the presence of disease, and the administration of therapeutic treatment. The aim of the present study was to identify optimal RGs in a set of blood samples collected at different time points (0, 24, 48, 72 h) from horses following administration of extracorporeal shock wave therapy (ESWT). The mRNA expression of twelve RGs: HPRT1, ACTB, HSP90A, SDHA, GUSB, B2M, UBC, NONO, TBP, H6PD, RPL32, GAPDH was determined using real time quantitative polymerase chain reaction (qPCR). An SAS program developed on the algorithm of geNorm, SASqPCR, was used to determine stability of the expression and the number of optimal RGs. The results showed that the range of quantification cycle (Cq) values of the evaluated genes varied between 17 and 26 cycles, and that one optimal RG, ACTB, was sufficient for normalization of gene expression. Results of stability of expression demonstrated that ACTB was the optimal choice for all the samples studied. Notably, in samples collected at 72 h post ESWT, TBP showed a significant change in the expression level, and was not suitable for use as a RG. These results substantiate the importance of validating and selecting an appropriate RG.展开更多
目的检测RFP(Ret finger protein)蛋白在人正常组织及人癌组织中的分布。方法采用实时荧光定量PCR方法,以18 S rRNA为内对照,检测RFP在人正常组织及人癌组织中的表达水平。结果RFP表达水平子宫颈鳞状细胞癌组织高于正常子宫颈组织,子宫...目的检测RFP(Ret finger protein)蛋白在人正常组织及人癌组织中的分布。方法采用实时荧光定量PCR方法,以18 S rRNA为内对照,检测RFP在人正常组织及人癌组织中的表达水平。结果RFP表达水平子宫颈鳞状细胞癌组织高于正常子宫颈组织,子宫内膜腺癌组织高于正常子宫内膜组织,胃腺癌组织高于正常胃组织,食管鳞状细胞癌组织高于正常食管组织,而子宫内膜腺癌组织高于子宫颈鳞状细胞癌组织,胃腺癌组织高于食管鳞状细胞癌组织,脑癌组织高于正常脑组织。结论RFP可能成为治疗恶性肿瘤的一个潜在的分子靶点。展开更多
Quantitative reverse transcription polymerase chain reaction(qRT-PCR) is the most commonly-used tool for measurement of gene expression, but its accuracy and reliability depend on appropriate data normalization with t...Quantitative reverse transcription polymerase chain reaction(qRT-PCR) is the most commonly-used tool for measurement of gene expression, but its accuracy and reliability depend on appropriate data normalization with the use of one or more stable reference genes. Adelphocoris suturalis is one of the most destructive pests of cotton, but until recently knowledge of its underlying molecular physiology had been hindered by a lack of molecular resources. To facilitate research on this pest, we evaluated 12 common housekeeping genes studied in insects(GAPDH, ACT, βACT, TBP, SDH, βTUB, EF1γ, EF1α, EF1δ, RPL32, RPS15, and RPL27) for their expression stability in A. suturalis when subjected to various experimental treatments, including three biotic(developmental stage and sex, tissue type, and metathoracic scent gland for varying developmental stages and sexes) and one abiotic(RNA interference injection) conditions. Four dedicated algorithms(ΔCt method, geNorm, BestKeeper and NormFinder) were used to analyze gene expression stability. In addition, RefFinder provided an overall ranking of the stability/suitability of these candidates. This study is the first to provide a comprehensive list of suitable reference genes for gene expression analyses in A. suturalis, which can serve to facilitate transcript expression study of related biological processes in this and related species.展开更多
Quantification of the BCR-ABL transcript is recommended to follow-up CML patients treated by imatinib mesylate (IM). Results are expressed as a normalized copy number (NCN) of BCR-ABL. We studied a cohort of 98 CML pa...Quantification of the BCR-ABL transcript is recommended to follow-up CML patients treated by imatinib mesylate (IM). Results are expressed as a normalized copy number (NCN) of BCR-ABL. We studied a cohort of 98 CML patients under IM as a first treatment and monitored by RQ-PCR after 12, 18 and 24 months according to the European LeukemiaNet recommendations. Our results support the hypothesis of an independent correlation between BCR-ABL NCN at diagnosis and major molecular response at 18 and 24 months in an inverse relationship. We also highlighted the possibility to use the NCN at diagnosis as a warning at diagnosis, and may be useful to identify patients who could benefit of a more rigorous follow-up.展开更多
文摘Selection of proper reference genes (RGs) is an essential step needed for accurate normalization of results from genomic studies. Expression of RGs is regulated by many factors such as species, age, gender, type of tissue, the presence of disease, and the administration of therapeutic treatment. The aim of the present study was to identify optimal RGs in a set of blood samples collected at different time points (0, 24, 48, 72 h) from horses following administration of extracorporeal shock wave therapy (ESWT). The mRNA expression of twelve RGs: HPRT1, ACTB, HSP90A, SDHA, GUSB, B2M, UBC, NONO, TBP, H6PD, RPL32, GAPDH was determined using real time quantitative polymerase chain reaction (qPCR). An SAS program developed on the algorithm of geNorm, SASqPCR, was used to determine stability of the expression and the number of optimal RGs. The results showed that the range of quantification cycle (Cq) values of the evaluated genes varied between 17 and 26 cycles, and that one optimal RG, ACTB, was sufficient for normalization of gene expression. Results of stability of expression demonstrated that ACTB was the optimal choice for all the samples studied. Notably, in samples collected at 72 h post ESWT, TBP showed a significant change in the expression level, and was not suitable for use as a RG. These results substantiate the importance of validating and selecting an appropriate RG.
文摘目的检测RFP(Ret finger protein)蛋白在人正常组织及人癌组织中的分布。方法采用实时荧光定量PCR方法,以18 S rRNA为内对照,检测RFP在人正常组织及人癌组织中的表达水平。结果RFP表达水平子宫颈鳞状细胞癌组织高于正常子宫颈组织,子宫内膜腺癌组织高于正常子宫内膜组织,胃腺癌组织高于正常胃组织,食管鳞状细胞癌组织高于正常食管组织,而子宫内膜腺癌组织高于子宫颈鳞状细胞癌组织,胃腺癌组织高于食管鳞状细胞癌组织,脑癌组织高于正常脑组织。结论RFP可能成为治疗恶性肿瘤的一个潜在的分子靶点。
基金The National Natural Science Foundation of China(31071775)The Science and Technology Department of Zhejiang Province(2008C24006)The Zhejiang Normal University Innovative Research Team Program
基金funded by the National Special Key Project for Transgenic Breeding, China (2016ZX08011002)the Open Fundation of State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences (SKLOF201415)
文摘Quantitative reverse transcription polymerase chain reaction(qRT-PCR) is the most commonly-used tool for measurement of gene expression, but its accuracy and reliability depend on appropriate data normalization with the use of one or more stable reference genes. Adelphocoris suturalis is one of the most destructive pests of cotton, but until recently knowledge of its underlying molecular physiology had been hindered by a lack of molecular resources. To facilitate research on this pest, we evaluated 12 common housekeeping genes studied in insects(GAPDH, ACT, βACT, TBP, SDH, βTUB, EF1γ, EF1α, EF1δ, RPL32, RPS15, and RPL27) for their expression stability in A. suturalis when subjected to various experimental treatments, including three biotic(developmental stage and sex, tissue type, and metathoracic scent gland for varying developmental stages and sexes) and one abiotic(RNA interference injection) conditions. Four dedicated algorithms(ΔCt method, geNorm, BestKeeper and NormFinder) were used to analyze gene expression stability. In addition, RefFinder provided an overall ranking of the stability/suitability of these candidates. This study is the first to provide a comprehensive list of suitable reference genes for gene expression analyses in A. suturalis, which can serve to facilitate transcript expression study of related biological processes in this and related species.
基金Supported by National Natural Science Foundation of China(No.81370477No.81370918)+2 种基金Science and Technology Research Outstanding Youth Fund of Hebei College(No.Y2011117)Undergraduate Innovation Project(No.201410081061No.X2014026)
文摘Quantification of the BCR-ABL transcript is recommended to follow-up CML patients treated by imatinib mesylate (IM). Results are expressed as a normalized copy number (NCN) of BCR-ABL. We studied a cohort of 98 CML patients under IM as a first treatment and monitored by RQ-PCR after 12, 18 and 24 months according to the European LeukemiaNet recommendations. Our results support the hypothesis of an independent correlation between BCR-ABL NCN at diagnosis and major molecular response at 18 and 24 months in an inverse relationship. We also highlighted the possibility to use the NCN at diagnosis as a warning at diagnosis, and may be useful to identify patients who could benefit of a more rigorous follow-up.
