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Knockdown of the atypical protein kinase genes GhABC1K2-A05 and GhABC1K12-A07 make cotton more sensitive to salt and PEG stress 被引量:1
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作者 Caixiang Wang Meili Li +3 位作者 Dingguo Zhang Xueli Zhang Juanjuan Liu Junji Su 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2024年第10期3370-3386,共17页
Activity of bc1 complex kinase(ABC1K)is an atypical protein kinase(aPK)that plays a crucial role in plant mitochondrial and plastid stress responses,but little is known about the responses of ABC1Ks to stress in cotto... Activity of bc1 complex kinase(ABC1K)is an atypical protein kinase(aPK)that plays a crucial role in plant mitochondrial and plastid stress responses,but little is known about the responses of ABC1Ks to stress in cotton(Gossypium spp.).Here,we identified 40 ABC1Ks in upland cotton(Gossypium hirsutum L.)and found that the Gh ABC1Ks were unevenly distributed across 17 chromosomes.The GhABC1K family members included 35 paralogous gene pairs and were expanded by segmental duplication.The GhABC1K promoter sequences contained diverse cis-acting regulatory elements relevant to hormone or stress responses.The qRT-PCR results revealed that most Gh ABC1Ks were upregulated by exposure to different stresses.Gh ABC1K2-A05 and Gh ABC1K12-A07 expression levels were upregulated by at least three stress treatments.These genes were further functionally characterized by virus-induced gene silencing(VIGS).Compared with the controls,the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced cotton lines exhibited higher malondialdehyde(MDA)contents,lower catalase(CAT),peroxidase(POD)and superoxide dismutase(SOD)activities and reduced chlorophyll and soluble sugar contents under NaCl and PEG stress.In addition,the expression levels of six stress marker genes(Gh DREB2A,Gh SOS1,Gh CIPK6,Gh SOS2,Gh WRKY33,and Gh RD29A)were significantly downregulated after stress in the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced lines.The results indicate that knockdown of Gh ABC1K2-A05 and Gh ABC1K12-A07 make cotton more sensitive to salt and PEG stress.These findings can provide valuable information for intensive studies of Gh ABC1Ks in the responses and resistance of cotton to abiotic stresses. 展开更多
关键词 COTTON ABC1K abiotic stress responses expression patterns virus-induced gene silencing(VIGS)
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The role of tazarotene-induced gene 1 in carcinogenesis:is it a tumor suppressor gene or an oncogene?
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作者 CHUN-HUA WANG LU-KAI WANG +1 位作者 RONG-YAUN SHYU FU-MING TSAI 《BIOCELL》 SCIE 2024年第9期1285-1297,共13页
Tazarotene-induced gene 1(TIG1)is induced by a derivative of vitamin A and is known to regulate many important biological processes and control the development of cancer.TIG1 is widely expressed in various tissues;yet... Tazarotene-induced gene 1(TIG1)is induced by a derivative of vitamin A and is known to regulate many important biological processes and control the development of cancer.TIG1 is widely expressed in various tissues;yet in many cancer tissues,it is not expressed because of the methylation of its promoter.Additionally,the expression of TIG1 in cancer cells inhibits their growth and invasion,suggesting that TIG1 acts as a tumor suppressor gene.However,in some cancers,poor prognosis is associated with TIG1 expression,indicating its protumor growth characteristics,especially in promoting the invasion of inflammatory breast cancer cells.This review comprehensively summarizes the roles of the TIG1 gene in cancer development and details the mechanisms through which TIG1 regulates cancer development,with the aim of understanding its various roles in cancer development. 展开更多
关键词 Tazarotene-induced gene 1 Retinoic acid receptor responder protein 1 Tumor suppressor gene ONCOGENE
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Candidate genes conferring ethylene-response in cultivated peanuts determined by BSA-seq and fine-mapping
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作者 Yanyan Tang Zhong Huang +6 位作者 Shaohui Xu Wenjie Zhou Jianjun Ren Fuxin Yu Jingshan Wang Wujun Ma Lixian Qiao 《The Crop Journal》 SCIE CSCD 2024年第3期856-865,共10页
Ethylene plays essential roles in plant growth,development and stress responses.The ethylene signaling pathway and molecular mechanism have been studied extensively in Arabidopsis and rice but limited in peanuts.Here,... Ethylene plays essential roles in plant growth,development and stress responses.The ethylene signaling pathway and molecular mechanism have been studied extensively in Arabidopsis and rice but limited in peanuts.Here,we established a sand-culture method to screen pingyangmycin mutagenized peanut lines based on their specific response to ethylene(“triple response”).An ethylene-insensitive mutant,inhibition of peanut hypocotyl elongation 1(iph1),was identified that showed reduced sensitivity to ethylene in both hypocotyl elongation and root growth.Through bulked segregant analysis sequencing,a major gene related to iph1,named AhIPH1,was preliminarily mapped at the chromosome Arahy.01,and further narrowed to a 450-kb genomic region through substitution mapping strategy.A total of 7014 genes were differentially expressed among the ACC treatment through RNA-seq analysis,of which only the Arahy.5BLU0Q gene in the candidate mapping interval was differentially expressed between WT and mutant iph1.Integrating sequence variations,functional annotation and transcriptome analysis revealed that a predicated gene,Arahy.5BLU0Q,encoding SNF1 protein kinase,may be the candidate gene for AhIPH1.This gene contained two single-nucleotide polymorphisms at promoter region and was more highly expressed in iph1 than WT.Our findings reveal a novel ethylene-responsive gene,which provides a theoretical foundation and new genetic resources for the mechanism of ethylene signaling in peanuts. 展开更多
关键词 Ethylene-insensitive Hypocotyl elongation AhIPH1 Candidate gene Genetic resources
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Pathogenesis of chronic enteropathy associated with the SLCO2A1 gene:Hypotheses and conundrums
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作者 Zhi-Xin Xie Yue Li +2 位作者 Ai-Ming Yang Dong Wu Qiang Wang 《World Journal of Gastroenterology》 SCIE CAS 2024年第19期2505-2511,共7页
Chronic enteropathy associated with the SLCO2A1 gene(CEAS)is a complex gastroenterological condition characterized by multiple ulcers in the small intestine with chronic bleeding and protein loss.This review explores ... Chronic enteropathy associated with the SLCO2A1 gene(CEAS)is a complex gastroenterological condition characterized by multiple ulcers in the small intestine with chronic bleeding and protein loss.This review explores the potential mechanisms underlying the pathogenesis of CEAS,focusing on the role of SLCO2A1-encoded prostaglandin transporter OATP2A1 and its impact on prostaglandin E2(PGE2)levels.Studies have suggested that elevated PGE2 levels contribute to mucosal damage,inflammation,and disruption of the intestinal barrier.The effects of PGE2 on macrophage activation and Maxi-Cl channel functionality,as well as its interaction with nonsteroidal anti-inflammatory drugs play crucial roles in the progression of CEAS.Understanding the balance between its protective and pro-inflammatory effects and the complex interactions within the gastrointestinal tract can shed light on potential therapeutic targets for CEAS and guide the development of novel,targeted therapies. 展开更多
关键词 SLCO2A1 Prostaglandin E2 Chronic enteropathy associated with the SLCO2A1 gene Small intestine MACROPHAGE
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New perspectives in prognostication of hepatocellular carcinoma:The role and clinical implications of transient receptor potential family genes
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作者 Shi-Hao Guan Wen-Jing Hu +2 位作者 Xin-Yu Wang Yue-Xia Gu De-Hua Zhou 《World Journal of Gastrointestinal Oncology》 SCIE 2024年第6期2862-2864,共3页
The study titled“Transient receptor potential-related risk model predicts prognosis of hepatocellular carcinoma patients”is a significant contribution to hepatocellular carcinoma(HCC)research,highlighting the role o... The study titled“Transient receptor potential-related risk model predicts prognosis of hepatocellular carcinoma patients”is a significant contribution to hepatocellular carcinoma(HCC)research,highlighting the role of transient receptor potential(TRP)family genes in the disease’s progression and prognosis.Utilizing data from The Cancer Genome Atlas database,it establishes a new risk assessment model,emphasizing the interaction of TRP genes with tumor proliferation pathways,key metabolic reactions like retinol metabolism,and the tumor immune microenvironment.Notably,the overexpression of the TRPC1 gene in HCC correlates with poorer patient survival outcomes,suggesting its potential as a prognostic biomarker and a target for personalized therapy,particularly in strategies combining immunotherapy and anti-TRP agents. 展开更多
关键词 Hepatocellular carcinoma Transient receptor potential channels TRPC1 gene Tumor immune microenvironment Cancer prognosis Bioinformatics in cancer research
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Understanding the role of transmembrane 9 superfamily member 1 in bladder cancer pathogenesis
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作者 Venkata Krishna Vamsi Gade Budhi Singh Yadav 《World Journal of Clinical Oncology》 2024年第4期468-471,共4页
In this editorial we comment on the article by Wei et al,published in the recent issue of the World Journal of Clinical Oncology.The authors investigated the role of Transmembrane 9 superfamily member 1(TM9SF1)protein... In this editorial we comment on the article by Wei et al,published in the recent issue of the World Journal of Clinical Oncology.The authors investigated the role of Transmembrane 9 superfamily member 1(TM9SF1)protein in bladder cancer(BC)carcinogenesis.Lentiviral vectors were used to achieve silencing or overexpression of TM9SF1 gene in three BC cell lines.These cell lines were then subject to cell counting kit 8,wound-healing assay,transwell assay,and flow cytometry.Proliferation,migration,and invasion of BC cells were increased in cell lines subjected to TM9SF1 overexpression.TM9SF1 silencing inhibited proliferation,migration and invasion of BC cells.The authors conclude that TM9SF1 may be an oncogene in bladder cancer pathogenesis. 展开更多
关键词 Urinary bladder cancer Transmembrane 9 superfamily member 1 gene cell line Lentiviral vectors Wound healing assay ONCOGENE Proliferation Migration
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A cluster of mutagenesis revealed an osmotic regulatory role of the OsPIP1 genes in enhancing rice salt tolerance 被引量:1
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作者 Leyuan Tao Bing Wang +6 位作者 Shichao Xin Wei Li Shengcai Huang Laihua Liu Jing Cui Qianru Zhang Xianguo Cheng 《The Crop Journal》 SCIE CSCD 2023年第4期1204-1217,共14页
Aquaporins play important regulatory roles in improving plant abiotic stress tolerance.To better understand whether the Os PIP1 genes collectively dominate the osmotic regulation in rice under salt stress,a cluster ed... Aquaporins play important regulatory roles in improving plant abiotic stress tolerance.To better understand whether the Os PIP1 genes collectively dominate the osmotic regulation in rice under salt stress,a cluster editing of the Os PIP1;1,Os PIP1;2 and Os PIP1;3 genes in rice was performed by CRISPR/Cas9 system.Sequencing showed that two mutants with Cas9-free,line 14 and line 18 were successfully edited.Briefly,line 14 deleted a single C base in both the Os PIP1;1 and Os PIP1;3 genes,and inserted a single T base in the Os PIP1;2 gene,respectively.While line 18 demonstrated an insertion of a single A base in the Os PIP1;1gene and a single T base in both the Os PIP1;2 and Os PIP1;3 genes,respectively.Multiplex editing of the Os PIP1 genes significantly inhibited photosynthetic rate and accumulation of compatible metabolites,but increased MDA contents and osmotic potentials in the mutants,thus delaying rice growth under salt stress.Functional loss of the Os PIP1 genes obviously suppressed the expressions of the Os PIP1,Os SOS1,Os CIPK24 and Os CBL4 genes,and increased the influxes of Na+and effluxes of K^(+)/H^(+)in the roots,thus accumulating more Na+in rice mutants under salt stress.This study suggests that the Os PIP1 genes are essential modulators collectively contributing to the enhancement of rice salt stress tolerance,and multiplex editing of the Os PIP1 genes provides insight into the osmotic regulation of the PIP genes. 展开更多
关键词 AQUAPORIN Multiplex gene editing CRISPR/Cas9 OsPIP1 genes Rice(Oryza sativa L.) Salt tolerance
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TLN1通过激活β-catenin信号通路负性调控骨髓间充质干细胞的成脂分化
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作者 黄红颜 柯皓腾 +5 位作者 叶伟佳 邓华宗 王涛 蔡明希 陈奇凡 岑水忠 《中国组织工程研究》 CAS 北大核心 2023年第10期1578-1583,共6页
背景:骨髓间充质干细胞成骨-成脂分化平衡在骨代谢疾病中发挥关键作用,课题组前期研究证实TLN1可调控骨髓间充质干细胞成骨分化,但其对骨髓间充质干细胞成脂分化的作用仍未明确。目的:探讨TLN1在骨髓间充质干细胞成脂分化中的作用及机... 背景:骨髓间充质干细胞成骨-成脂分化平衡在骨代谢疾病中发挥关键作用,课题组前期研究证实TLN1可调控骨髓间充质干细胞成骨分化,但其对骨髓间充质干细胞成脂分化的作用仍未明确。目的:探讨TLN1在骨髓间充质干细胞成脂分化中的作用及机制。方法:诱导骨髓间充质干细胞成脂分化并使用Western blot及qRT-PCR检测TLN1的表达变化;采用CCK-8检测TLN1敲减后骨髓间充质干细胞增殖能力变化,成脂诱导后进行油红O染色及定量分析,qRT-PCR检测骨髓间充质干细胞成脂分化的关键分子PPAR-γ、C/EBP-α的基因表达水平;Western blot检测成脂经典通路蛋白的表达,同时在敲减TLN1的同时添加β-catenin激活剂后检测骨髓间充质干细胞的成脂分化能力,以探讨TLN1调节成脂分化的具体分子机制。结果与结论:TLN1在骨髓间充质干细胞的成脂分化过程中表达下调;敲减TLN1对骨髓间充质干细胞的增殖无明显影响,可显著增强骨髓间充质干细胞成脂诱导分化能力;敲减TLN1可抑制活性的β-catenin表达,而激活β-catenin信号通路可逆转TLN1敲减介导的成脂分化增强。结果表明,TLN1表达在骨髓间充质干细胞成脂分化过程中逐渐下调,敲减TLN1通过抑制β-catenin信号通路发挥促进骨髓间充质干细胞成脂分化的作用。 展开更多
关键词 间充质干细胞 tln1 成脂分化 Β-CATENIN 信号通路 组织工程
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High-resolution genetic mapping and identification of candidate genes for the wheat stem rust resistance gene Sr8155B1
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作者 Jian Wang Hongyu Li +13 位作者 Tao Shen Shikai Lyu Shams ur Rehman Hongna Li Guiping Wang Binyang Xu Qing Wang Wanyi Hu Kairong Li Shengsheng Bai Jian Ma Haitao Yu Matthew N.Rouse Shisheng Chen 《The Crop Journal》 SCIE CSCD 2023年第6期1852-1861,共10页
Stem rust,caused by Puccinia graminis f.sp.tritici(Pgt),threatens global wheat production.Development of cultivars with increased resistance to stem rust by identification,mapping,and deployment of resistance genes is... Stem rust,caused by Puccinia graminis f.sp.tritici(Pgt),threatens global wheat production.Development of cultivars with increased resistance to stem rust by identification,mapping,and deployment of resistance genes is the best strategy for controlling the disease.In this study,we performed fine mapping and characterization of the all-stage stem rust resistance(Sr)gene Sr8155B1 from the durum wheat line 8155-B1.In seedling tests of biparental populations,Sr8155B1 was effective against six Chinese Pgt races tested.In a segregating population of 5060 gametes,Sr8155B1 was mapped to a 0.06-cM region flanked by markers Pku2772 and Pku43365,corresponding to 1.5-and 2.7-Mb regions in the Svevo and Chinese Spring reference genomes.Both regions include several typical nucleotide-binding leucine-rich repeat(NLR)and protein kinase genes that represent candidate genes.Among them,three NLR genes and three receptor-like protein kinases were highly polymorphic between the parental lines and their transcripts were upregulated in the homozygous resistant line TdR2 relative to its susceptible sister line TdS4.Four markers(Pku2772,Pku43365,Pku2950,and Pku3721)developed in this study,together with seedling resistance responses,correctly predicted Sr8155B1 absence or presence in 78 tetraploid wheat genotypes tested.The presence of Sr8155B1 in tetraploid wheat accessions CItr 14916,PI 197492,and PI 197493 was confirmed by mapping in three F_(2)populations.The genetic map and linked markers developed in this study may accelerate the deployment of Sr8155B1-mediated resistance in wheat breeding programs. 展开更多
关键词 Durum wheat Stem rust Resistance gene Sr8155B1 CC-NBS-LRR
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Regulatory role of NFAT1 signaling in articular chondrocyteactivities and osteoarthritis pathogenesis
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作者 MINGCAI ZHANG TANNER CAMPBELL +1 位作者 SPENCER FALCON JINXI WANG 《BIOCELL》 SCIE 2023年第10期2125-2132,共8页
Osteoarthritis (OA), the most common form of joint disease, is characterized clinically by joint pain, stiffness,and deformity. OA is now considered a whole joint disease;however, the breakdown of the articular cartil... Osteoarthritis (OA), the most common form of joint disease, is characterized clinically by joint pain, stiffness,and deformity. OA is now considered a whole joint disease;however, the breakdown of the articular cartilage remains themajor hallmark of the disease. Current treatments targeting OA symptoms have a limited impact on impeding orreversing the OA progression. Understanding the molecular and cellular mechanisms underlying OA development isa critical barrier to progress in OA therapy. Recent studies by the current authors’ group and others have revealedthat the nuclear factor of activated T cell 1 (NFAT1), a member of the NFAT family of transcription factors, regulatesthe expression of many anabolic and catabolic genes in articular chondrocytes of adult mice. Mice lacking NFAT1exhibit normal skeletal development but display OA in both appendicular and spinal facet joints as adults. Thisreview mainly focuses on the recent advances in the regulatory role of NFAT1 transcription factor in the activities ofarticular chondrocytes and its implication in the pathogenesis of OA. 展开更多
关键词 OSTEOARTHRITIS CHONDROCYTE NFAT1 Transcription factor Regulation of gene expression
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应用基因芯片筛选胃癌淋巴转移相关基因及TLN1的初步研究 被引量:1
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作者 张黎 赵仲生 +2 位作者 马杰 茹国庆 董来华 《中国病理生理杂志》 CAS CSCD 北大核心 2009年第7期1271-1278,共8页
目的:应用基因表达谱芯片筛选胃癌正常胃黏膜、胃癌原发灶和淋巴结转移灶的差异表达基因,并结合生物信息学方法筛选胃癌淋巴结转移相关基因,探讨TLN1与胃癌淋巴结转移的关系。方法:应用Affy-metrix芯片筛选癌旁正常胃黏膜(A组)、胃癌原... 目的:应用基因表达谱芯片筛选胃癌正常胃黏膜、胃癌原发灶和淋巴结转移灶的差异表达基因,并结合生物信息学方法筛选胃癌淋巴结转移相关基因,探讨TLN1与胃癌淋巴结转移的关系。方法:应用Affy-metrix芯片筛选癌旁正常胃黏膜(A组)、胃癌原发灶(B组)和淋巴结转移灶(C组)的差异表达基因,利用队列试验的基因表达谱分析、基于聚类分析结果的基因功能(基于Gene Ontology分类)集群分析和Pathway分析等生物信息学方法对结果进行分析,并用RT-PCR检测TLN1在胃癌原发灶和淋巴结转移灶的表达情况。结果:A、B、C3组呈连续上调的差异表达基因278个,其功能主要集中在免疫应答、细胞黏附、磷酸盐转运、阴离子转移、细胞趋化和移动、信号转导等方面;连续下调的387个基因其功能主要集中在消化、糖类代谢、脂质代谢、蛋白代谢、一氧化氮复合物代谢、一氧化碳复合物代谢、细胞黏附等方面。Pathway分析显示在整合素介导的细胞黏附信号通路异常激活,THBS1、TLN1、CAPN3、ITGAX、SORBS1、CAPN6、CAPN9在3组中呈连续梯度改变。TLN1在癌旁非肿瘤胃黏膜、胃癌原发灶和淋巴结转移灶表达量分别为:0.0000342±0.0000711、0.1064±0.1251和0.2886±0.3529;在胃癌淋巴结转移灶显著高表达(P<0.05)。结论:基因芯片技术结合生物信息学方法筛选胃癌淋巴转移相关基因,有助于找到胃癌淋巴道转移的关键基因或通路,为寻找肿瘤淋巴道转移的早期诊断指标及新的治疗靶点奠定了实验基础,本研究结果初步表明在整合素信号通路中扮演重要角色的TLN1基因异常表达与胃癌淋巴结转移密切相关。 展开更多
关键词 胃肿瘤 基因 tln1 肿瘤转移 基因表达芯片
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Changes of p53 and Waf1p21 and cell proliferation in esophageal carcinogenesis 被引量:13
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作者 WANG Li Dong 1, YANG Wan Cai 1, ZHOU Qi 1, XING Ying 1,JIA Yun Ying 2 and ZHAO Xin 1 《World Journal of Gastroenterology》 SCIE CAS CSCD 1997年第2期30-32,共3页
AIM To study the correlationship between the changes of p53, Waf1p21 and the cell proliferation determined by PCNA at different stages of human esophageal carcinogenesis. METHODS Biopsied and resected esophageal ti... AIM To study the correlationship between the changes of p53, Waf1p21 and the cell proliferation determined by PCNA at different stages of human esophageal carcinogenesis. METHODS Biopsied and resected esophageal tissues from a high risk population for esophageal cancer in northern China were used in this study. All the specimens were fixed with 85% alcohol and further processed with routine histology. The avidin biotin peroxidase complex (ABC) method was used for the detection of p53, Waf1p21 and PCNA. RESULTS The strong nuclear staining for p53, Waf1p21 and PCNA was observed in the normal esophageal epithelium and the epithelia with different severities of lesions. As the lesions progressed to dysplasia (DYS) and to esophageal squamons cell carcinoma (SCC), the percentage of Waf1p21 immunoreactivity decreased. The number of Waf1p21 immunostaining positive cells increased slightly from normal to basal cell hyperplasia (BCH), but there was no further increase in DYS and in SCC. The total number of positive cells for Waf1p21 stain appeared to be lower than that of p53 in normal and BCH esophageal epithelia and much lower in DYS and SCC. The Waf1p21 positive immunostaining cells were located at the third and forth cell layers in half of the samples examined, which was 2~4 cell layers higher than that of PCNA and p53 in the same histological categories of normal, BCH and DYS. CLNCLUSION The low levels of Waf1p21 at the stage of DYS may be related to a functional loss of p53. Other mechanisms may also be responsible to the lack of Waf1p21 expression in DYS and SCC. 展开更多
关键词 ESOPHAGEAL neoplasms PRECANCEROUS conditions P53 genes Waf1p21 genes suppressor tumor
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大鼠Neurogenesin-1基因真核表达载体的构建及在cos-7细胞中的表达 被引量:2
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作者 高维陆 尹宗生 +1 位作者 张胜权 张辉 《安徽医科大学学报》 CAS 北大核心 2009年第4期430-433,共4页
目的克隆大鼠海马中Neurogenesin-1(Ng1)基因片段,构建pSecTag2/HygroB-Ng1真核表达载体,并检测其在cos-7细胞中的表达,为进一步研究该基因对脊髓神经干细胞分化的影响提供实验依据。方法在无RNA酶污染的条件下提取出大鼠海马总RNA。利... 目的克隆大鼠海马中Neurogenesin-1(Ng1)基因片段,构建pSecTag2/HygroB-Ng1真核表达载体,并检测其在cos-7细胞中的表达,为进一步研究该基因对脊髓神经干细胞分化的影响提供实验依据。方法在无RNA酶污染的条件下提取出大鼠海马总RNA。利用逆转录聚合酶链反应扩增出Ng1基因片段。将该基因片段连接到真核表达载体pSecTag2/HygroB,聚合酶链反应初步筛选,双酶切鉴定后送测序。将构建成功的重组真核表达载体转染入cos-7细胞,Westernblot鉴定重组Ng1蛋白的表达。结果逆转录聚合酶链反应成功获得大鼠Ng1cDNA。随机挑选10个重组真核表达载体的克隆,聚合酶链反应筛选出阳性克隆2个,经双酶切鉴定、测序及Blast分析鉴定重组质粒构建成功。脂质体介导转染cos-7细胞48h后,Westernblot鉴定重组Ng1蛋白在cos-7细胞中的表达,在46ku处出现阳性条带。结论大鼠海马Ng1基因的真核表达载体pSecTag2/HygroB-Ng1构建成功,转染cos-7细胞后能够表达重组Ng1蛋白。 展开更多
关键词 大鼠 基因表达 遗传载体 Neurogenesin-1
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NS特异性干扰载体pGenesil-1-NS的构建及鉴定
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作者 高鸿霞 王国庆 +2 位作者 刘燕 霍中华 李钰 《北华大学学报(自然科学版)》 CAS 2015年第6期729-732,共4页
目的核干细胞因子(nucleostemin,NS)是维持干细胞和癌细胞增殖所必需的蛋白质,可能成为肿瘤基因治疗的潜在靶点.本文旨在构建靶向NS干扰载体p Genesil-1-NS,为后续实验奠定基础.方法基于已发布的NS mRNA序列(NM_206825),选取5'-AAGC... 目的核干细胞因子(nucleostemin,NS)是维持干细胞和癌细胞增殖所必需的蛋白质,可能成为肿瘤基因治疗的潜在靶点.本文旨在构建靶向NS干扰载体p Genesil-1-NS,为后续实验奠定基础.方法基于已发布的NS mRNA序列(NM_206825),选取5'-AAGCCTA GGAAAGACCCAGG-3'(397-416)作为候选靶序列,按shRNA载体设计原则,设计合成两条互补DNA链,退火后插入载体,并以酶切和测序鉴定重组转化子.结果经酶切及测序鉴定证实合成序列正确插入载体.结论成功构建NS特异性干扰载体p Genesil-1-NS,可用于NS在肿瘤中的功能研究. 展开更多
关键词 核干细胞因子基因 pgenesil-1载体 发夹状RNA RNA干扰
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Study on the Function of ORF Genes of Porcine Circovirus-like Virus P1
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作者 Libin WEN Xuejiao ZHU +2 位作者 Qi XIAO Wei WANG Kongwang HE 《Agricultural Biotechnology》 CAS 2021年第2期84-88,92,共6页
[Objectives]This study was conducted to determine the functions of eight ORF genes of porcine circovirus-like virus P1.[Methods]The double-copy tandem molecular cloning of porcine circovirus-like virus P1 genome was u... [Objectives]This study was conducted to determine the functions of eight ORF genes of porcine circovirus-like virus P1.[Methods]The double-copy tandem molecular cloning of porcine circovirus-like virus P1 genome was used to construct molecular clones with eight ORFs deleted by DNA site-directed mutagenesis technology.After transfected into PK15 cells for a certain period of time,RNA were extracted and was used to verify whether the eight ORFs were deleted or not and used for gene microarry analysis.The GO functions and KEGG pathway enrichment of differentially expressed genes were analyzed.[Results]P1 ORF1 is mainly involved in the biological processes of defense response to virus,signal transduction,regulation of Rab GTPase activity,and lipid metabolic process,and involved in the molecular functions of protein phosphatase inhibitor activity,phosphatidylinositol phospholipase C activity,2 iron,2 sulfur cluster binding,phosphoric diester hydrolase activity,and Rab GTPase activator activity,and in the KEGG pathways of secretion of digestive gland and nervous system development.P1 ORF2 is mainly involved in the biological processes of positive regulation of leukocyte chemotaxis,positive regulation of cell proliferation,positive regulation of cell migration,defense response to virus,regulation of cell growth,and involved in the molecular functions of insulin-like growth factor binding,and chemokine activity,and in the KEGG pathways of cytosolic DNA-sensing pathway,RIG-I-like receptor signaling pathway,toll-like receptor signaling pathway,chemokine signaling pathway,and cytokines,cytokine-cytokine receptor interaction.The biological processes,molecular functions and related pathways involving P1 ORF3 and ORF5 are basically similar to those of ORF2.P1 ORF8 is mainly involved in the biological processes of purine ribonucleotide biosynthetic process,amino acid transport,defense response to virus,amino acid transmembrane transport,and involved in molecular functions of N6-(1,2-dicarboxyethyl)AMP AMP-lyase(fumarate-forming)activity,iron-sulfur cluster binding,amino acid transmembrane transporter activity.[Conclusions]The analysis of the ORF functions of P1 virus lays a foundation for the study of its pathogenicity and pathogenesis. 展开更多
关键词 Porcine circovirus-like virus P1 Function of ORF genes MICROARRAY Differentially expressed genes
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TLN1-PI3K/AKT通路调控骨髓间充质干细胞成骨分化 被引量:4
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作者 李兆峰 王鹏 +3 位作者 谢中瑜 岑水忠 李进腾 沈慧勇 《岭南现代临床外科》 2019年第4期387-391,共5页
目的探讨Talin1在骨髓间充质干细胞(Mesenchymal stem cell,MSC)成骨分化中的作用及其调控机制。方法诱导MSC成骨分化后通过qRT-PCR及Western blot检测TLN1的表达变化,在MSC中敲低TLN1后采用CCK-8检测细胞增殖能力改变,利用ALP活性检测... 目的探讨Talin1在骨髓间充质干细胞(Mesenchymal stem cell,MSC)成骨分化中的作用及其调控机制。方法诱导MSC成骨分化后通过qRT-PCR及Western blot检测TLN1的表达变化,在MSC中敲低TLN1后采用CCK-8检测细胞增殖能力改变,利用ALP活性检测及茜素红染色观察成骨分化变化情况。通过Western blot实验研究TLN1在MSC成骨分化过程中参与的分子机制。结果qPCR及Western blot结果提示,TLN1在成骨分化过程中表达上升;siRNA敲低TLN1表达后,MSC增殖无明显差异,ALP活性及矿化结节定量较对照组下降(P<0.05)。Western blot结果显示敲低TLN1表达后p-AKT表达下降,回补实验证明激活PI3K-AKT表达可回复MSC的成骨分化能力。结论诱导MSC成骨分化后TLN1表达逐渐上调,敲低TLN1后通过PI3K-AKT通路抑制骨髓间充质干细胞成骨分化。 展开更多
关键词 骨髓间充质干细胞 tln1 成骨分化 PI3K-AKT
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Cotton Plants Transformed with the Activated Chimeric Cry1Ac and API-B Genes 被引量:12
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作者 郭洪年 吴家和 +6 位作者 陈晓英 罗晓丽 卢睿 石跃进 秦红敏 肖娟丽 田颖川 《Acta Botanica Sinica》 CSCD 2003年第1期108-113,共6页
A chimeric gene, Bt29K, composed of coding sequences of activated Cry1Ac insecticidal protein and an endoplasm reticulum-retarding signal peptide, was synthesized. A plant expression vector containing two expression c... A chimeric gene, Bt29K, composed of coding sequences of activated Cry1Ac insecticidal protein and an endoplasm reticulum-retarding signal peptide, was synthesized. A plant expression vector containing two expression cassettes for the Bt29K and API-B genes was constructed. These two insect-resistant genes were transferred into two cotton ( Gossypium hirsutum L.) varieties ( or lines) via Agrobacterium-mediated transformation and nine homozygous transgenic cotton lines showing a mortality of 90.0% - 99.7% to cotton ballworm (Heliothis armigera) larvae and good agronomic traits were selected through six generations. Molecular biology analysis revealed that one or two copies of the insecticidal protein genes were integrated into the transgenic cotton genome and activated Cry1Ac and API-B protein expression was at a level of 0.17% and 0.09% of the total soluble protein in the transgenic cotton leaves, respectively. Comparison of the insect-resistance of the homozygous lines expressing the activated chimeric Cry1Ac and API-B with that expressing Cry1Ac only revealed that the insect-resistance of the former is apparently higher than the latter. These results also indicate that the strategy to construct a plant expression vector expressing two different insect-resistant genes reported here is reasonable. 展开更多
关键词 synthetic chimeric Cry1Ac gene arrowhead proteinase inhibitor gene insect-resistant transgenic cotton plants
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The Role of Predominant Expression of Th2 Type Cytokines Gene in the Genesis and Development of Human Gliomas 被引量:1
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作者 李刚 胡永生 +3 位作者 李新钢 张庆林 贾德泽 宫崧峰 《The Chinese-German Journal of Clinical Oncology》 CAS 2003年第4期227-230,252,253,共6页
Objective: To explore the expression of Th1/Th2 cytokines gene in human gliomas and its role in the genesis and development of human gliomas.Methods: Using IL-2 and IFNγ as Th1 type cytokines, IL-4, IL-6 and IL-10 as... Objective: To explore the expression of Th1/Th2 cytokines gene in human gliomas and its role in the genesis and development of human gliomas.Methods: Using IL-2 and IFNγ as Th1 type cytokines, IL-4, IL-6 and IL-10 as Th2 type cytokines, the biological activity of cytokines in the supernatant of glioma cell lines was assayed by ELISA method, and the gene expression of Th1/Th2 cytokines in human glioma cells, glioma infiltrating lymphocytes and glioma cell lines were detected by RT-PCR.Results: There was predominant expression of Th2 type cytokines in human glioma cells, glioma infiltrating lymphocytes and glioma cell lines, but there was no such expression in normal brain tissues.Conclusion: It suggested that there is a relationship between the Th2 type cytokines expression in human gliomas and the immunosupressive status of human glioma patients. The predominant expression of Th2 type cytokines may play an important role in the genesis and development of human gliomas. Key words glioma - Th1/Th2 - gene expression - RT-PCR This project was supported by a grant from National Natural Sciences foundation of China (No. 30271335). 展开更多
关键词 GLIOMA TH1/TH2 gene expression RT-PCR
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The Effects of Dwarfing Genes (Rht-B1b, Rht-D1b, and Rht8) with Different Sensitivity to GA_3 on the Coleoptile Length and Plant Height of Wheat 被引量:14
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作者 TANG Na JIANG Ying +1 位作者 HE Bei-ru HU Yin-gang 《Agricultural Sciences in China》 CAS CSCD 2009年第9期1028-1038,共11页
Understanding the effects of wheat dwarfing genes on the coleoptile length and plant height is crucial for the proper utilization of dwarfing genes in the improvement of wheat yield. Molecular marker analysis combined... Understanding the effects of wheat dwarfing genes on the coleoptile length and plant height is crucial for the proper utilization of dwarfing genes in the improvement of wheat yield. Molecular marker analysis combined with pedigree information were used to classify wheat cultivars widely planted in major wheat growing regions in China into different categories based on the dwarfing genes they carried. The effects of the dwarfing genes with different sensitivity to gibberellins (GA3) on the coleoptile length and plant height were analyzed. Screening of 129 cultivars by molecular marker analysis revealed that 58 genotypes of wheat contained the dwarfing gene Rht-B1b, 24 genotypes of wheat contained Rht-D1b gene and 73 genotypes of wheat possessed Rht8 gene. In addition, among these 129 cultivars, 35 genotypes of wheat cultivars contained both Rht-B1b and Rht8 genes and 16 genotypes of wheat cultivars contained both Rht-D1b and Rht8 genes. Wheat cultivars with the dwarfing genes Rht-B1b or Rht-D1b were insensitive to GA3, while the cultivars with the dwarfing gene Rht8 were sensitive to GA3. Most of the wheat genotypes containing combination of Rht8 gene with either Rht-B1b or Rht-D1b gene were insensitive to GA3. The plant height was reduced by 24.6, 30.4, 28.2, and 32.2%, respectively, for the wheat cultivars containing Rht-B1b, Rht-D1b, Rht-B1b + Rht8, and Rht-D1b + Rht8 genes. The plant height was reduced by 14.3% for the wheat cultivar containing GA3-sensitive gene Rht8. The coleoptile length was shortened by 25.4, 31.3, 28.4 and 31.3%, respectively, in the wheat cultivars containing Rht-B1b, Rht-D1b, Rht-B1b +Rht8 and Rht-D1b + Rht8 genes, while the coleoptile length was shortened only by 6.2% for the wheat cultivar containing Rht8 gene. We conclude that GA3-insensitive dwarfing genes (Rht-B1b and Rht-D1b) are not suitable for the wheat improvement in dryland because these two genes have effect on reducing both plant height and coleoptile length. In contrast, GA3- sensitive dwarfing gene (Rht8) is a relatively ideal candidate for the wheat improvement since it significantly reduces the plant height of wheat, but has less effect on the coleoptile length. 展开更多
关键词 bread wheat dwarfing genes Rht-B1b Rht-D1b Rht8 coleoptile length plant height
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EMP-1 Promotes Tumorigenesis of NSCLC through PI3K/AKT Pathway 被引量:2
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作者 来森艳 王桂华 +3 位作者 曹小年 李兆明 胡俊波 王晶 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2012年第6期834-838,共5页
This study examined the role of EMP-1 in tumorigenesis of non-small cell lung carcinoma (NSCLC) and the possible mechanism. Specimens were collected from 28 patients with benign lung diseases and 28 with NSCLC, and im... This study examined the role of EMP-1 in tumorigenesis of non-small cell lung carcinoma (NSCLC) and the possible mechanism. Specimens were collected from 28 patients with benign lung diseases and 28 with NSCLC, and immunohis to chemically detected to evaluate the correlation of EMP-1 expression to the clinical features of NSCLC. Recombinant adenovirus was constructed to over-express EMP-1 and then infect PC9 cells. Cell proliferation was measured by Ki67 staining. Western blotting was performed to examine the effect of EMP-1 on the PI3K/AKT signaling. Moreover, tumor xeno-grafts were established by subcutaneous injection of PC9 cell suspension (about 5×107/mL in 100 μL of PBS) into the right hind limbs of athymic nude mice. The results showed EMP-1 was significantly up-regulated in NSCLC patients as compared with those with benign lung diseases. Over-expression of EMP-1 promoted proliferation of PC9 cells, which coincided with the activation of the PI3K/AKT pathway. EMP-1 promoted the growth of xenografts of PC9 cells in athymic nude mice. It was concluded that EMP-1 expression may contribute to the development and progress of NSCLC by activating PI3K/AKT pathway. 展开更多
关键词 NSCLC EMP-1 gene PI3K/AKT pathway TUMORIgenesIS
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