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Myeloidcell-lineage and premylocytic-stage-specific-expression of the mouse myeloperoxidase gene is controlled at initiation as well as elongation levels of transcription 被引量:2
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作者 ZHU JING DE (Department of Medical Oncology, University of Glasgow,Glasgow G61 1BD, UK) 《Cell Research》 SCIE CAS CSCD 1999年第2期107-134,共28页
The myeloperoxidase (MPO) is an important microbicidal protein present at high concentration in the primary granule of mature granulocyte and its expression is regulated in both myeloidcell-lineage and premyelocytic-s... The myeloperoxidase (MPO) is an important microbicidal protein present at high concentration in the primary granule of mature granulocyte and its expression is regulated in both myeloidcell-lineage and premyelocytic-stagespecific manners. A better understanding of the underlying control mechanisms should provide insights into the temporal and co-ordinate regulation of the gene expression during granulopoiesis. We have identified its promoter by mapping the start(s) of transcription using various molecular approaches together with demonstrating the promoter function of the relevant DNA segment in a transient transfection reporter assay. Besides the major start of transcription mapped at G residue, 11 nucleotide upstream of the 3’ end of exon 0, the usage of that is specific to the MPO expressing cell lines, we have shown that irrespective of the MPO-expression status of the hematopoietic cells, transcription occurs broadly within a two kb region upstream of the 5’ proximity of the gene, and is largely terminated in nitron 2. These data support a model of the pre myelocytic-stage-specific MPO expression, the control of which is operated at initiation as well as elongation levels of transcription. 展开更多
关键词 Myeloid-cell-lineage premylocytic-stage-specific-expression myloperosidase gene gene controll
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Hypoxic response elements and Tet-On advanced double-controlled systems regulate hVEGF_(165) and angiopoietin-1 gene expression in vitro 被引量:1
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作者 Hao Zhang Hongyan Dong +4 位作者 Bo Jiang Zheng Wang Rui Chen Zhifeng Zhang Zhongming Zhang 《The Journal of Biomedical Research》 CAS 2011年第3期204-212,共9页
Angiogenesis in ischemic tissue is a complex and multi-gene event. In the study, we constructed hypoxic re-sponse elements (HRE) and the Tet-On advanced double-controlled systems and investigated their effects on th... Angiogenesis in ischemic tissue is a complex and multi-gene event. In the study, we constructed hypoxic re-sponse elements (HRE) and the Tet-On advanced double-controlled systems and investigated their effects on the expression of hVEGF165 and angiopoietin-1 (Ang-1) genes in rat cardiomyocytes exposed to hypoxia and pharma-cologic induction. We infected neonatal rat cardiomyocytes with recombinant rAAV-rtTA-Rs-M2/rAAV-TRE-Tight-Ang-1 and rAAV-9HRE- hVEGF165. Our results indicated that the viral titer was 1×1012 vg /mL and the viral purity exceeded 98%. hVEGF165 expression was induced by hypoxia, but not by normoxia (P 0.001). Ang-1 expression was evident under doxycycline induction, but undetectable without doxycycline induction (P 0.001). Immunofluorescence staining showed that positively stained hVEGF165 and Ang-1 protein appeared only under both hypoxia and doxycycline induction. We demonstrate here that HRE and the recombinant Tet-On advanced double gene-controlled systems sensitively regulate the expression of hVEGF165 and Ang-1 genes in an altered oxygen environment and under pharmacological induction in vitro. 展开更多
关键词 gene control vascular endothelial growth factor ANGIOPOIETIN-1 hypoxia DOXYCYCLINE CARDIOMYOCYTE
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The Genetic Structure and Diversity of Repomucenus curvicornis Inhabiting Liaoning Coast Based on Mitochondrial COⅠ Gene and Control Region
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作者 Li Yulong Liu Xiuze +3 位作者 Yu Xuguang Li Yiping Fu Jie Dong Jing 《Animal Husbandry and Feed Science》 CAS 2018年第1期12-17,共6页
[Object] This study was conducted to explore the genetic diversity and structure of the wild Repomucenus curvicornis inhabiting Liaoning Coast, China. [Method] The mitochondrial COⅠ gene and control region(CR) were... [Object] This study was conducted to explore the genetic diversity and structure of the wild Repomucenus curvicornis inhabiting Liaoning Coast, China. [Method] The mitochondrial COⅠ gene and control region(CR) were PCR amplified from the wild R. curvicornis populations from the Liaodong Bay(n=22) and the northern Yellow Sea(n=18), sequenced and analyzed for genetic diversity. [Result] The contents of A, T, C and G of 624 bp COⅠ gene were 24.09%, 31.04%, 25.28%, and 19.59%, and those of 460 bp CR fragment were 32.96%, 32.80%, 14.86% and 19.38%, respectively. The total number of variable sites, average number of nucleotide differences( k), haplotype diversity(H) and nucleotide diversity(π) based on COⅠ gene were 38, 4.67,(0.96±0.02) and(0.007 5±0.004 2), and those based on CR fragment were 26, 3.35,(0.97 ±0.02) and(0.007 3±0.004 3), respectively. Based on mitochondrial COⅠ gene and CR, the genetic diversity of Liaodong Bay population was lower than that of the northern Yellow Sea population. The AMOVA analysis based on CR fragments revealed almost significant genetic divergence between the Liaodong Bay and the northern Yellow Sea populations, while there was no significant genetic divergence based on COⅠ gene. The results showed that CR and COⅠ gene are effective molecular markers for detecting the genetic diversity of R. curvicornis population, while CR is more reliable than COⅠ gene in detecting the genetic structure. [Conclusion] CR is an appropriate marker for genetic analysis of marine fish population. 展开更多
关键词 Repomucenus curvicornis Mitochondrial DNA COⅠ gene control region sequence genetic diversity genetic differentiation
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Validation of housekeeping genes as internal controls for studying the gene expression in Pyropia haitanensis(Bangiales, Rhodophyta) by quantitative real-time PCR 被引量:6
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作者 LI Bing CHEN Changsheng +2 位作者 XU Yan JI Dehua XIE Chaotian 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2014年第9期152-159,共8页
Pyropia haitanensis is an economically important mariculture crop in China and has a high research value for several life phenomena, for example environmental tolerance. To explore the mechanisms underlying these char... Pyropia haitanensis is an economically important mariculture crop in China and has a high research value for several life phenomena, for example environmental tolerance. To explore the mechanisms underlying these characteristics, gene expression has been investigated at the whole transcriptome level. Gene expression studies using quantitative real-time PCR should start by selecting an appropriate internal control gene; therefore, the absolute expression abundance of six housekeeping genes (18S rRNA (18S), ubiquitin-conju-ating enzyme (UBC), actin (ACT), β-tubulin (TUB), elongation factors 2 (EF2), and glyceraldehyde-3-phos- phate dehydrogenase (GAPDH) examined by the quantitative real-time PCR in samples corresponding to different strains, life-cycle stages and abiotic stress treatments. Their expression stabilities were assessed by the comparative cycle threshold (Ct) method and by two different software packages: geNorm and NormFinder. The most stable housekeeping gene is UBC and the least stable housekeeping is GADPH. Thus, it is proposed that the most appropriate internal control gene for expression analyses in P. haitanensis is UBC. The results pave the way for further gene expression analyses of different aspects of P. haitanensis biology including different strains, life-history stages and abiotic stress responses. 展开更多
关键词 Pyropia haitanensis quantitative real-time PCR internal control genes gene expression
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Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury 被引量:4
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作者 Wei Liu Jie Yu +2 位作者 Yi-Fan Wang Qian-Qian Shan Ya-Xian Wang 《Neural Regeneration Research》 SCIE CAS CSCD 2022年第6期1387-1392,共6页
There is a lack of systematic research on the expression of internal control genes used for gene expression normalization in real-time reverse transcription polymerase chain reaction in spinal cord injury research.In ... There is a lack of systematic research on the expression of internal control genes used for gene expression normalization in real-time reverse transcription polymerase chain reaction in spinal cord injury research.In this study,we used rat models of spinal cord hemisection to analyze the expression stability of 13 commonly applied reference genes:Actb,Ankrd27,CypA,Gapdh,Hprt1,Mrpl10,Pgk1,Rictor,Rn18s,Tbp,Ubc,Ubxn11,and Ywhaz.Our results show that the expression of Ankrd27,Ubc,and Tbp were stable after spinal cord injury,while Actb was the most unstable internal control gene.Ankrd27,Ubc,Tbp,and Actb were consequently used to investigate the effects of internal control genes with differing stabilities on the normalization of target gene expression.Target gene expression levels and changes over time were similar when Ankrd27,Ubc,and Tbp were used as internal controls but different when Actb was used as an internal control.We recommend that Ankrd27,Ubc,and Tbp are used as internal control genes for real-time reverse transcription polymerase chain reaction in spinal cord injury research.This study was approved by the Administration Committee of Experimental Animals,Jiangsu Province,China(approval No.20180304-008)on March 4,2018. 展开更多
关键词 geNorm analysis reference genes internal control genes NORMALIZATION NormFinder analysis reverse transcription-quantitative polymerase chain reaction spinal cord injury stability of gene expression
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Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis 被引量:1
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作者 王霞 冯建华 +3 位作者 黄爱优 何林文 牛建峰 王广策 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2017年第6期1432-1441,共10页
Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the... Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal Rhodophyta. Here, the effects of salinity and light intensity on the quantum yield of photosystem II in Py. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under different stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The cDNA was synthesized and the expression levels of seven potential internal control genes were evaluated using qRT-PCR method. Then, we used geNorm, a common statistical algorithm, to analyze the qRT-PCR data of seven reference genes. Potential genes that may constantly be expressed under different conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while tubulin, glyceraldehyde- 3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the geNorm screening. Based on the results of the software analysis and absolute quantification, we proposed that elongation factor 3 and 18S ribosomal RNA could be used as internal control genes when the Py. haitanensis blades were subjected to salinity stress, and that a-tubulin and 18S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis. 展开更多
关键词 real-time quantitative PCR housekeeping genes internal control genes stress responding Pyropia haitanensis
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Identification of stable internal control genes for accurate normalization of real-time quantitative PCR data in testicular tissue from two breeds of cattle
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作者 Pradeep Nag Ankur Sharma +6 位作者 Elango Kamaraj Arumugam Kumaresan Tirtha Kumar Datta Ayyasamy Manimaran Nilendu Paul Sakthivel Jeyakumar Kerekoppa P.Ramesha 《Asian pacific Journal of Reproduction》 2020年第5期247-255,共9页
Objective:To assess the stability of 10 candidate internal control genes(ICGs),namely GAPDH,ACTB,RPL23,RPS15A,ATPSF1,GLUT5,HMBS,ATP2B4,PPIA,and BRP to normalize the transcriptional data from testes samples of Zebu and... Objective:To assess the stability of 10 candidate internal control genes(ICGs),namely GAPDH,ACTB,RPL23,RPS15A,ATPSF1,GLUT5,HMBS,ATP2B4,PPIA,and BRP to normalize the transcriptional data from testes samples of Zebu and crossbred bulls.Methods:Total RNA was isolated from testicular tissue of Zebu and crossbred bulls(n=6 each)between 2-8 years of age.cDNA was synthesized,and the quantitative real-time polymerase chain reaction(PCR)was performed.The cycle threshold values were used for the analysis of the stability of ICGs.Four different statistical algorithms:geNorm,Normfinder,BestKeeper,and RefFinder,were used to assess the stability of these genes.Results:ATPSF1,HMBS,PPIA,and RPS15A were the most reliable and stable ICGs for Zebu testes,and ATPSF1,RPL23,and PPIA for crossbred testes.Conclusions:A panel of stable ICGs(ATPSF1,HMBS,PPIA,RPS15A for Zebu and ATPSF1,RPL23,and PPIA for crossbred)for normalization of gene expression data in testes samples can be helpful for researchers to conduct functional genomics studies at the testicular level in cattle bulls. 展开更多
关键词 Cattle bulls gene expression Internal control genes NORMALIZATION qPCR Testis
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Construction and Control of Genetic Regulatory Networks:A Multivariate Markov Chain Approach
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作者 Shu-Qin Zhang Ling-Yun Wu +2 位作者 Wai-Ki Ching Yue Jiao Raymond, H. Chan 《Journal of Biomedical Science and Engineering》 2008年第1期15-21,共7页
In the post-genomic era, the construction and control of genetic regulatory networks using gene expression data is a hot research topic. Boolean networks (BNs) and its extension Probabilistic Boolean Networks (PBNs) h... In the post-genomic era, the construction and control of genetic regulatory networks using gene expression data is a hot research topic. Boolean networks (BNs) and its extension Probabilistic Boolean Networks (PBNs) have been served as an effective tool for this purpose. However, PBNs are difficult to be used in practice when the number of genes is large because of the huge computational cost. In this paper, we propose a simplified multivariate Markov model for approximating a PBN The new model can preserve the strength of PBNs, the ability to capture the inter-dependence of the genes in the network, qnd at the same time reduce the complexity of the network and therefore the computational cost. We then present an optimal control model with hard constraints for the purpose of control/intervention of a genetic regulatory network. Numerical experimental examples based on the yeast data are given to demonstrate the effectiveness of our proposed model and control policy. 展开更多
关键词 gene Expression SEQUENCES MULTIVARIATE MARKOV CHAIN Optimal control Policy Probabilistic BOOLEAN Networks.
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Advances in Research and Application of gE Gene/Protein in Prevention and Control of Swine Pseudora-bies
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作者 Ma Li Yang Limei +6 位作者 Zhuang Jinqiu Xu Qianqian Wang Yan Guo Shijin Shen Zhiqiang Wang Yanping Zhang Ying 《Animal Husbandry and Feed Science》 CAS 2017年第2期93-95,100,共4页
Research and application progresses of gE gene and its encoding gE protein in PR vaccines, diagnostic technique and epidemiological investigation are summarized, which have certain reference value for comprehensive pr... Research and application progresses of gE gene and its encoding gE protein in PR vaccines, diagnostic technique and epidemiological investigation are summarized, which have certain reference value for comprehensive prevention and control of PR and gradual purification of PR in different regions. 展开更多
关键词 Pseudorabies virus (PRV) gE gene gE protein VACCINE DIAGNOSIS Epidemiological investigation Prevention and control
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番茄褐色皱纹果病毒研究进展
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作者 秦会琴 王银磊 +6 位作者 赵丽萍 宋刘霞 董舒超 陈杰 曹凤妍 孙敏 赵统敏 《江苏农业学报》 北大核心 2026年第1期207-216,共10页
番茄褐色皱纹果病毒(Tomato brown rugose fruit virus,ToBRFV)是烟草花叶病毒属(Tobamovirus)中新发现的一种RNA病毒。该病毒于2014年首次在以色列被鉴定,随后在全球范围内快速扩散,目前已在亚洲、欧洲、美洲和非洲的多个国家和地区广... 番茄褐色皱纹果病毒(Tomato brown rugose fruit virus,ToBRFV)是烟草花叶病毒属(Tobamovirus)中新发现的一种RNA病毒。该病毒于2014年首次在以色列被鉴定,随后在全球范围内快速扩散,目前已在亚洲、欧洲、美洲和非洲的多个国家和地区广泛分布,对番茄、辣椒等重要茄科作物的安全生产构成了严重威胁。与烟草花叶病毒属的其他成员相比,ToBRFV在运动蛋白区域具有独特的分子特征,使其能够突破番茄中常见的Tm-2等抗病毒基因,表现出更强的侵染能力。本文系统梳理了近年来国内外关于ToBRFV的研究进展,从ToBRFV的发生与传播、结构特征、检测鉴定、抗病研究、综合防治等研究进展方面进行了归纳总结,以期为人们认识病毒、有效防治病毒病提供参考。 展开更多
关键词 番茄 番茄褐色皱纹果病毒 抗病基因 综合防治
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抗生素抗性菌的适应性代价及其防控策略研究进展
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作者 罗裔宵 陈洁蓉 +6 位作者 罗智聪 郑昕晨 罗俊金 黎玥 温馨 陈涛 吴银宝 《微生物学报》 北大核心 2026年第2期595-609,共15页
微生物的抗生素抗性(antibiotic resistance,AR)问题已成为全球公共卫生安全面临的重大挑战。微生物获得AR后常伴随适应性代价,通常在无抗生素环境中其群落竞争力弱于敏感菌,这使得抗生素抗性基因(antibiotic resistance genes,ARGs)成... 微生物的抗生素抗性(antibiotic resistance,AR)问题已成为全球公共卫生安全面临的重大挑战。微生物获得AR后常伴随适应性代价,通常在无抗生素环境中其群落竞争力弱于敏感菌,这使得抗生素抗性基因(antibiotic resistance genes,ARGs)成为细菌的负担。基于此,本文详细阐述了适应性代价的产生机制与测量方法,举例说明了不同获取方式下ARGs介导的适应性代价,着重介绍了染色体介导和质粒介导的ARGs在适应性代价方面的差异及分子机制,最后提出了基于适应性代价的抗生素抗性菌(antibiotic resistance bacteria,ARB)防控策略。本文揭示了抗生素抗性与细菌适应性之间的权衡生物学规律,为解决由ARB及ARGs引发的全球公共卫生安全问题提供了防控思路。 展开更多
关键词 抗生素抗性菌 适应性代价 抗性机制 水平基因转移 防控
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TRP家族蛋白的环境感知功能和炎症调控效应及其在水生动物中的研究进展
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作者 汪蕾 李苡萱 +3 位作者 由凯文 郭钰琪 吴亚梅 谢海平 《水生生物学报》 北大核心 2026年第1期201-215,共15页
环境中的物理、化学和生物因素与动物的生命活动密切相关,感知这些因素的变化对动物避免有害刺激或转移至适宜栖息地至关重要。瞬时受体电位通道(Transient receptor potential,TRP)蛋白属于阳离子通道家族,它使膜电位去极化并调节细胞... 环境中的物理、化学和生物因素与动物的生命活动密切相关,感知这些因素的变化对动物避免有害刺激或转移至适宜栖息地至关重要。瞬时受体电位通道(Transient receptor potential,TRP)蛋白属于阳离子通道家族,它使膜电位去极化并调节细胞内阳离子(如Ca^(2+))的浓度,不仅介导动物对环境刺激的感知,还深度参与免疫炎症反应等多种生理过程,故也被称为“生物传感器”。文章首先对TRP蛋白家族的结构、分类进行综述性介绍,聚焦于其温度感知与炎症调控功能的最新进展。在此基础上,深入总结TRP家族蛋白在水生动物环境适应(温度、机械、化学感知)及生理调控(摄食行为、免疫防御、繁殖)中的关键角色,探讨TRP蛋白家族作为药物靶点和在水生动物疾病防控与育种的潜力,为提高水产养殖行业的健康发展提供有益参考。 展开更多
关键词 TRP蛋白家族 离子通道 温度感知 炎症反应 药物开发 疾病防控 水生动物
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鸭疫里默氏杆菌的防治
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作者 潘志强 王丽媛 于辰龙 《家禽科学》 2026年第1期40-44,共5页
鸭疫里默氏杆菌(Riemerella anatipestifer,RA)是危害水禽养殖业的重要病原菌,隶属于黄杆菌科,现有25个血清型,血清型间缺乏交叉免疫保护。该菌对高温及常用消毒剂敏感,易被灭活,但在潮湿环境中可长期存活。RA在全球肉鸭、蛋鸭养殖场广... 鸭疫里默氏杆菌(Riemerella anatipestifer,RA)是危害水禽养殖业的重要病原菌,隶属于黄杆菌科,现有25个血清型,血清型间缺乏交叉免疫保护。该菌对高温及常用消毒剂敏感,易被灭活,但在潮湿环境中可长期存活。RA在全球肉鸭、蛋鸭养殖场广泛流行,其毒力基因则直接调控病原菌的感染性与致病性,死亡率介于5%~70%间,给养禽业造成严重经济损失。当前RA耐药性问题日益突出,已检测到四环素类、氨基糖苷类、氯霉素类等多种耐药基因。为控制RA的流行与危害,亟需构建基于耐药机制与致病机制研究的综合防治体系,以有效控制疾病传播蔓延。 展开更多
关键词 鸭疫里默氏杆菌 耐药基因 毒力基因 防控
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枯草芽孢杆菌J6-1对桃褐腐病菌的抑菌活性分析
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作者 李晓倩 王卓妮 +5 位作者 史冰柯 王丽 周贝贝 涂洪涛 袁洪波 侯珲 《园艺学报》 北大核心 2026年第1期229-243,共15页
为防治由果生链核盘菌(Monilinia fructicola)引起的桃褐腐病,采用平皿对峙法筛选出拮抗细菌枯草芽孢杆菌J6-1,并分析其对桃褐腐病的防治效果和拮抗机制。结果显示,菌株J6-1对桃褐腐病菌的抑制率达到74%,且具有广谱抑菌活性。菌株J6-1... 为防治由果生链核盘菌(Monilinia fructicola)引起的桃褐腐病,采用平皿对峙法筛选出拮抗细菌枯草芽孢杆菌J6-1,并分析其对桃褐腐病的防治效果和拮抗机制。结果显示,菌株J6-1对桃褐腐病菌的抑制率达到74%,且具有广谱抑菌活性。菌株J6-1能产生蛋白酶、纤维素酶、葡聚糖酶等水解酶,其上清液可以破坏菌丝细胞膜,诱导桃果实病程相关基因(PpPR1)的表达。2%、4%、6%、8%的J6-1上清液对桃褐腐病菌菌丝生长的抑制率分别为64.3%、70.2%、79.75%、85.43%,抑制能力随浓度升高而逐渐增强,且2%的J6-1上清液能引起桃褐腐病菌菌丝变细、消解,原生质外渗,菌丝细胞膜受损。J6-1粗提物中差异幅度最大的前10个差异代谢物主要是氨基酸及其衍生物。桃果实接种M.fructicola 3 d进行菌株J6-1菌悬液、上清液与粗提物处理,抑制效果分别为73.81%、82.48%、92.66%。表明菌株J6-1对桃褐腐病具有良好的防治效果。 展开更多
关键词 褐腐病 枯草芽孢杆菌 生物防治 抑制效果 病程相关基因
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MTHFR 677 C/T基因检测质控品制备及性能评估
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作者 孙其濛 巴红平 +4 位作者 潘利萍 程娅 郑莹 陈永刚 龚洁 《临床检验杂志》 2026年第1期54-58,共5页
目的制备性能稳定、均匀性好、适用性广且分别含有MTHFR 677位点野生型(CC)、突变型(TT)和杂合型(CT)的质控品,并对其性能进行评估,为开展MTHFR 677基因检测室间质量评价,评估参评实验室MTHFR 677位点基因检测能力及存在问题提供实验依... 目的制备性能稳定、均匀性好、适用性广且分别含有MTHFR 677位点野生型(CC)、突变型(TT)和杂合型(CT)的质控品,并对其性能进行评估,为开展MTHFR 677基因检测室间质量评价,评估参评实验室MTHFR 677位点基因检测能力及存在问题提供实验依据。方法利用PCR和Sanger测序技术从200例人抗凝全血的临床样本中筛选出分别含有MTHFR 677位点野生型(CC)、突变型(TT)和杂合型(CT)的DNA样本,并依据CNAS-GL03《能力验证样品均匀性和稳定性评价指南》中关于均匀性和稳定性评价的检测方法,对筛选的样本进行均稳性验证。通过均稳性验证后,将筛选的DNA样本按30 ng/μL的浓度进行稀释并以30μL/管进行分装,分别作为MTHFR 677位点野生型、突变型和杂合型质控品。制备含有MTHFR 677位点3种基因型不同组合的5支质控盲样,选择6家已开展叶酸检测工作的公立医院,采取6家医院各自使用的检测方法和检测试剂,对5支盲样质控品进行质量验证。结果从200例人抗凝全血临床样本中提取全血DNA,经Sanger测序后筛选到分别含有MTHFR 677位点野生型(CC)、突变型(TT)和杂合型(CT)的DNA样本;采用实时荧光定量PCR对样本进行均匀性和稳定性检测,测得的MTHFR 677位点基因型与预期基因型符合率为100%;6家公立医院对5支质控盲样检测结果正确率为100%。结论本研究制备的MTHFR 677 C/T基因检测质控品均匀性、稳定性和适用性良好,为使用人抗凝全血临床样本制备MTHFR 677位点基因检测质控品提供了实验依据。 展开更多
关键词 亚甲基四氢叶酸还原酶 全血DNA 基因检测 质量控制
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造血干细胞基因治疗产品的研发应用和质量控制
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作者 陈悦 华君男 刘超 《中国细胞生物学学报》 2026年第1期219-233,共15页
造血干细胞(HSCs)凭借其自我更新能力与多谱系分化潜能,已成为细胞和基因治疗领域的重要工具。造血干细胞基因治疗(HSC-GT)通过对患者自体HSCs进行体外基因修饰并回输,可从病因层面纠正疾病,实现“一次治疗、终身获益”的治疗目标,同时... 造血干细胞(HSCs)凭借其自我更新能力与多谱系分化潜能,已成为细胞和基因治疗领域的重要工具。造血干细胞基因治疗(HSC-GT)通过对患者自体HSCs进行体外基因修饰并回输,可从病因层面纠正疾病,实现“一次治疗、终身获益”的治疗目标,同时规避异体造血干细胞移植中供体配型困难及移植物抗宿主病等风险。该文系统梳理了HSC-GT的技术发展历程,包括从γ-逆转录病毒载体的初步尝试,到第三代自灭活型慢病毒载体在安全性与效率上的突破,再到CRISPR-Cas9、碱基编辑及先导编辑等技术推动的精准基因修饰革命。临床转化方面,HSC-GT领域已有多款创新药物在欧盟和美国获批上市(包括全球首款CRISPR编辑药物),其适应症已从血红蛋白病等血液系统疾病,逐步拓展至溶酶体贮积症和中枢神经系统疾病等非血液领域,展现出广阔的应用前景。质量研究与控制是HSC-GT产品开发及生产的核心环节,需围绕造血干细胞特性、基因修饰功效、细胞纯度、安全性四个维度,贯彻“质量源于设计”理念,遵循药品生产质量管理规范,建立过程控制和终端质量放行标准,从而确保产品的安全性、有效性与质量可控性。展望未来,HSC-GT将朝着体内基因修饰、适应症拓展以及更精准的质量评价体系等方向持续发展,有望从高端个体化治疗手段逐步转化为更可及、更可负担的医疗产品,为多种难治性疾病提供新的治疗选择。 展开更多
关键词 造血干细胞 基因治疗 病毒载体 基因编辑 质量控制 临床应用
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腕踝针治疗带状疱疹后神经痛临床疗效及对血清NaV1.7、CGRP含量的影响
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作者 刘伟群 张锦锈 +3 位作者 陈晓 蔡建荣 林新强 魏真 《中国针灸》 北大核心 2026年第2期199-205,共7页
目的:观察腕踝针治疗带状疱疹后神经痛(PHN)的临床疗效及对患者血清电压门控钠通道1.7(NaV1.7)、降钙素基因相关肽(CGRP)含量的影响。方法:将90例PHN患者随机分为腕踝针组(45例,脱落2例)和西药组(45例,脱落1例,中止1例)。西药组予局部外... 目的:观察腕踝针治疗带状疱疹后神经痛(PHN)的临床疗效及对患者血清电压门控钠通道1.7(NaV1.7)、降钙素基因相关肽(CGRP)含量的影响。方法:将90例PHN患者随机分为腕踝针组(45例,脱落2例)和西药组(45例,脱落1例,中止1例)。西药组予局部外用5%利多卡因凝胶贴膏;腕踝针组予腕踝针治疗,每次30 min,每日1次,连续5 d为一疗程,疗程间休息2 d。两组均治疗4周。分别于治疗前、治疗2周后、治疗4周后及治疗结束后4周随访观察两组患者疼痛视觉模拟量表(VAS)、匹兹堡睡眠质量指数(PSQI)、焦虑自评量表(SAS)和抑郁自评量表(SDS)评分,于治疗前、治疗4周后检测两组患者血清NaV1.7、CGRP含量,记录两组患者试验期间应急用药及不良反应发生情况,并于治疗4周后评定临床疗效。结果:两组患者治疗2、4周后及随访期疼痛VAS、PSQI、SAS、SDS评分均较治疗前降低(P<0.05);治疗4周后、随访时,腕踝针组患者上述评分低于西药组(P<0.05)。治疗4周后,两组患者血清NaV1.7、CGRP含量均较治疗前降低(P<0.05),且腕踝针组低于西药组(P<0.05)。试验期间腕踝针组服用应急镇痛药物5例,西药组服用应急镇痛药物11例,两组比较差异无统计学意义(P>0.05);腕踝针组患者不良反应发生率为9.3%,低于西药组的27.9%(P<0.05)。腕踝针组总有效率为90.7%(39/43),西药组总有效率为83.7%(36/43),两组总有效率比较差异无统计学意义(P>0.05)。结论:腕踝针可有效治疗PHN,降低患者疼痛程度、改善睡眠质量及负性情绪,其机制可能与下调血清NaV1.7、CGRP含量有关。 展开更多
关键词 带状疱疹后神经痛 腕踝针 针刺 电压门控钠通道1.7(NaV1.7) 降钙素基因相关肽(CGRP) 随机对照试验
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Characterization of reference genes for qPCR analysis in various tissues of the Fujian oyster Crassostrea angulata 被引量:2
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作者 濮菲 杨丙晔 柯才焕 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2015年第4期838-845,共8页
Accurate quantification of transcripts using quantitative real-time polymerase chain reaction (qPCR) depends on the identification of reliable reference genes for normalization. This study aimed to identify and vali... Accurate quantification of transcripts using quantitative real-time polymerase chain reaction (qPCR) depends on the identification of reliable reference genes for normalization. This study aimed to identify and validate seven reference genes, including actin-2 (ACT-2), elongation factor 1 alpha (EF-1α), elongation factor 1 beta (EF-1β), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ubiquitin (UBQ), β-tubulin (β-TUB), and 18 S ribosomal RNA, from Crassostrea angulata, a valuable marine bivalve cultured worldwide. Transcript levels of the candidate reference genes were examined using qPCR analysis and showed differential expression patterns in the mantle, gill, adductor muscle, labial palp, visceral mass, hemolymph and gonad tissues. Quantitative data were analyzed using the geNorm software to assess the expression stability of the candidate reference genes, revealing that β-TUB and UBQ were the most stable genes. The commonly used GAPDH and 18S rRNA showed low stability, making them unsuitable candidates in this system. The expression pattern of the G protein β-subunit gene (Gβ) across tissue types was also examined and normalized to the expression of each or both of UBQ andβ-TUB as internal controls. This revealed consistent trends with all three normalization approaches, thus validating the reliability of UBQ and β-TUB as optimal internal controls. The study provides the first validated reference genes for accurate data normalization in transcript profiling in Crassostrea angulata, which will be indispensable for further fimetional genomics studies in this economically valuable marine bivalve. 展开更多
关键词 Crassostrea angulata gene expression quantitative real-time PCR internal control gene G protein β-subunit gene
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细胞与基因治疗产品中慢病毒载体生产与质控要求
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作者 陈成 刘晶晶 +2 位作者 卢京兰 程海子 吴钦斌 《中国细胞生物学学报》 2026年第1期184-194,共11页
慢病毒载体(lentiviral vectors,LVs)因其能够高效感染分裂和非分裂细胞,并实现外源基因的长期、稳定表达,已成为细胞与基因治疗领域最常见的基因递送工具之一,尤其在CAR-T细胞治疗和遗传病治疗方面具有广泛应用。该文旨在系统综述慢病... 慢病毒载体(lentiviral vectors,LVs)因其能够高效感染分裂和非分裂细胞,并实现外源基因的长期、稳定表达,已成为细胞与基因治疗领域最常见的基因递送工具之一,尤其在CAR-T细胞治疗和遗传病治疗方面具有广泛应用。该文旨在系统综述慢病毒载体的生产与质控要求。该文回顾了慢病毒载体系统的历史发展脉络,包括其从HIV-1病毒改造而来的发展历程、四代系统的安全性演进,并对比了国内外在该领域的技术发展路径与监管框架;系统阐述了当前慢病毒载体生产的主流策略,包括基于贴壁细胞和悬浮细胞的规模化生产工艺、瞬时转染与稳定生产细胞系构建的优缺点,以及下游的纯化与浓缩策略;重点讨论了慢病毒载体的质量控制体系,涵盖生物学活性(滴度)、纯度、安全性[无菌、支原体、复制型慢病毒(RCL)等]和理化特性等关键指标的检测方法与标准。最后,展望了慢病毒载体未来发展方向,包括生产技术的创新(如稳定细胞系开发、无血清悬浮培养工艺)、分析检测方法(新一代测序技术、快速检测方法)的优化以及规模化生产中难题的解决,以期为推动基于慢病毒载体的细胞与基因治疗产品的临床转化与产业化提供参考。 展开更多
关键词 慢病毒载体 细胞与基因治疗 生产工艺 质量控制 复制型慢病毒
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Association of the Apolipoprotein B Gene Polymorphisms With Essential Hypertension in Northern Chinese Han Population 被引量:5
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作者 WEI-YAN ZHAO JIAN-FENG HUANG +5 位作者 LAI-YUAN WANG HONG-FAN LI PENG-HUA ZHANG QI ZHAO SHU-FENG CHEN DONG-FENG GU 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2007年第3期260-264,共5页
Objective To study the association of the apolipoprotein B gene polymorphisms with essential hypertension in Northern Chinese Han population. Methods XbaI and EcoRl polymorphisms of the apolipoprotein B (APOB) gene ... Objective To study the association of the apolipoprotein B gene polymorphisms with essential hypertension in Northern Chinese Han population. Methods XbaI and EcoRl polymorphisms of the apolipoprotein B (APOB) gene were genotyped by polymerase chain reaction (PCR) and restriction fragment-length polymorphism (RFLP) method in 503 unrelated hypertensive patients and 490 healthy controls recruited from international collaborative study of cardiovascular disease in Asia (InterAsia). Results The difference in the genotypic distributions could be neglected across the groups. The prevalence of X+ allele in healthy controls (4.8%) was less frequent in Chinese, and there was no significant difference in the frequency of the X+ allele between cases (5.7%) and controls (P=0.38). The observed E- allele frequencies were closely similar among groups (5.9% in cases vs 5.0% in controls, P=0.39). Logitstic regression analyses revealed that the lack of association still persisted after adjustment of other environmental factors. Haplotype analysis showed that X-E+ was most frequent and no haplotype could significantly contribute to essential hypertension. Conclusion The APOB gene XbaI and EcoRI polymorphisms are not associated with essential hypertension in the Northern Chinese Han population. Future studies on single nucleotide polymorphisms in larger samples are needed to further investigate the possible contribution of the APOB gene to essential hypertension. 展开更多
关键词 Essential hypertension APOB gene POLYMORPHISMS Case-control study
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