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ISG15蛋白在原发性肝细胞癌中表达及其对预后的影响
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作者 黄丽芳 宋建华 +3 位作者 韦翠容 林子芹 杨达平 粟连秀 《现代医药卫生》 2024年第2期209-212,共4页
目的探讨干扰素刺激基因15(ISG15)蛋白在原发性肝细胞癌(HCC)中的表达水平及其临床预后价值。方法选取2018年1月至2020年10月该院经术后病理确诊的HCC患者100例,根据ISG15蛋白表达水平将其分为低表达组和高表达组。比较不同临床特征患者... 目的探讨干扰素刺激基因15(ISG15)蛋白在原发性肝细胞癌(HCC)中的表达水平及其临床预后价值。方法选取2018年1月至2020年10月该院经术后病理确诊的HCC患者100例,根据ISG15蛋白表达水平将其分为低表达组和高表达组。比较不同临床特征患者ISG15蛋白表达水平,分析不同ISG15蛋白表达水平患者生存情况。结果癌组织及癌旁组织中ISG15蛋白表达水平分别为(5.00±1.69)、(3.78±1.22),二者比较,差异有统计学意义(t=5.766,P<0.001)。不同肿物大小、HBV-DNA拷贝数及分化程度患者ISG15蛋白表达水平比较,差异有统计学意义(P<0.05)。低表达组中位生存期为40.5个月,生存率为70.83%,高表达组中位生存期为33.0个月,生存率为24.39%,二者比较,差异有统计学意义(χ^(2)=9.980,P=0.0016)。结论ISG15蛋白在HCC癌组织中高表达,其在抑制肿瘤细胞生长、抵抗HBV病毒复制、抑制癌细胞分化过程中具有相关作用。ISG15蛋白表达与临床预后相关,低表达者生存期更长。 展开更多
关键词 原发性肝细胞癌 干扰素刺激基因15 预后
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新西兰白兔BMP 15基因的真核表达载体构建、表达模式及其在卵巢组织的表达
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作者 陈孟娟 刘雨晴 +4 位作者 王智通 温佳乐 许会芬 于光晴 李明 《畜牧兽医学报》 CAS CSCD 北大核心 2024年第2期562-575,共14页
旨在获得新西兰白兔BMP 15基因序列及其组织表达规律,构建其真核表达载体,预测BMP15的生物学功能。本研究选择180日龄性成熟的健康新西兰白兔作为研究对象,采用RT-PCR技术扩增BMP 15基因CDS区序列,利用T4连接方法将目的片段连接至线性化... 旨在获得新西兰白兔BMP 15基因序列及其组织表达规律,构建其真核表达载体,预测BMP15的生物学功能。本研究选择180日龄性成熟的健康新西兰白兔作为研究对象,采用RT-PCR技术扩增BMP 15基因CDS区序列,利用T4连接方法将目的片段连接至线性化的Pmcherry-N1和pCMV-Myc空载体,构建真核表达载体,利用生物信息学分析其编码蛋白的性质及结构。采用实时荧光定量PCR(qRT-PCR)技术与Western blotting技术检测pCMV-Myc-BMP 15过表达情况。接下来,使用qRT-PCR检测BMP 15基因在不同组织中的表达水平。激光共聚焦方法检测BMP 15基因的亚细胞定位。利用免疫荧光技术检测内源BMP15在新西兰白兔卵巢组织定位情况。结果表明:克隆得到新西兰白兔BMP 15基因CDS区序列长1182 bp,PCR及测序结果表明pCMV-Myc-BMP 15和Pmcherry-N1-BMP 15真核表达载体构建成功。生物信息学分析显示,BMP 15基因编码393个氨基酸;BMP15蛋白不稳定系数为55.32,等电点大小为9.69,是一种稳定的碱性蛋白质。BMP15蛋白共有26个磷酸化位点,15个糖基化位点,存在信号肽,不存在跨膜结构域。系统进化树表明,新西兰白兔与猪亲缘关系最近,与鸡的亲缘关系最远。二级结构和三级结构分析结果表明,BMP15蛋白主要由α-螺旋(38.68%)、无规则卷曲(37.4%)、延伸链(15.52%)、β-转角(8.40%)组成,为混合型蛋白。蛋白互作关系预测发现,BMP15蛋白与FSHR、FIGLA、BMPR1B、AMHR2、NOBOX等卵巢生长发育的相关蛋白之间存在相互作用。不同组织表达分析显示,BMP 15基因在卵巢组织中特异性表达;激光共聚焦结果显示,BMP15主要存在于在细胞质中。将pCMV-Myc-BMP 15转染至HEK293T细胞内,BMP15的mRNA水平和蛋白水平均显著上调。卵巢组织免疫荧光检测结果显示,BMP15主要定位在卵巢颗粒细胞的细胞质中。本研究成功构建了BMP15的真核表达载体,对BMP 15基因及其编码的蛋白的理化性质和生物学特性进行了预测分析,在HEK293T细胞内成功过表达,并得到了该基因的亚细胞定位情况、组织表达情况及在卵巢组织的分布情况。为后续开展BMP 15基因在卵巢生长发育中的功能及机制研究提供了理论依据。 展开更多
关键词 新西兰白兔 BMP 15基因 克隆 组织表达 细胞定位
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长链非编码RNA CASC15对肝细胞癌增殖、侵袭和迁移的影响及作用机制
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作者 吴东洋 蔡青山 +4 位作者 刘东 左刚刚 李树栋 刘立友 郑建兴 《实用肿瘤学杂志》 CAS 2024年第1期18-29,共12页
目的探究长链非编码RNA(lncRNA)癌症易感性基因15(Cancer susceptibility candidate 15,CASC15)在肝细胞癌(Hepatocellular carcinoma,HCC)中的分子调控机制。方法通过生物信息学方法预测目的基因表达,并分析目的基因表达与患者生存时... 目的探究长链非编码RNA(lncRNA)癌症易感性基因15(Cancer susceptibility candidate 15,CASC15)在肝细胞癌(Hepatocellular carcinoma,HCC)中的分子调控机制。方法通过生物信息学方法预测目的基因表达,并分析目的基因表达与患者生存时间的关系;收集临床HCC患者肝癌组织和癌旁组织;CCK-8、Transwell和流式细胞术实验检测HCC细胞SMMC7721和Huh-7的增殖、侵袭、迁移以及凋亡;双荧光素酶实验检测miR-144-3p与CASC15和富含亮氨酸的重复序列蛋白1(Leucine rich repeat containing protein 1,LRRC1)的靶向关系;qRT-PCR和Western blot检测mRNA和蛋白表达情况;免疫荧光实验用于蛋白定位研究;回复实验验证CASC15/miR-144-3p/LRRC1对HCC进展的影响。体内实验验证CASC15对HCC进展的影响。结果TCGA数据库与qRT-PCR检测显示HCC组织和细胞中CASC15高表达、miR-144-3p低表达、LRRC1高表达(P<0.05)。增殖、侵袭、迁移的细胞功能实验结果表明在肿瘤发展中CASC15和LRRC1起到促进作用,miR-144-3p则是抑制作用,与凋亡实验结果一致(P<0.05)。细胞功能实验表明CASC15抑制miR-144-3p功能,miR-144-3p抑制LRRC1,CASC15与miR-144-3p结合,并导致LRRC1上调。回复实验结果表明CASC15通过抑制miR-144-3p促进LRRC1的表达从而促进HCC细胞增殖、侵袭和迁移并抑制细胞凋亡。结论CASC15可能通过调节miR-144-3p/LRRC1轴,从而促进HCC进展。 展开更多
关键词 癌症易感性基因15 miR-144-3p 富含亮氨酸的重复序列蛋白1 肝癌细胞 细胞生物学功能
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肿瘤相关基因ADAM15在结肠癌中一个非同义变异对细胞黏附及侵袭的影响
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作者 王晶 贺礼兵 +2 位作者 张国燕 刘筱涵 程杉 《首都医科大学学报》 北大核心 2023年第6期1044-1052,共9页
目的解析ADAM15基因单核苷酸变异(single nucleotide variant,SNV)在结肠癌肿瘤发生发展相关的细胞学过程中的具体功能作用。方法首先对具有侵袭表型的结肠癌患者癌组织及癌旁组织全外显子组测序,然后进行野生型/突变型ADAM15结肠癌HCT-... 目的解析ADAM15基因单核苷酸变异(single nucleotide variant,SNV)在结肠癌肿瘤发生发展相关的细胞学过程中的具体功能作用。方法首先对具有侵袭表型的结肠癌患者癌组织及癌旁组织全外显子组测序,然后进行野生型/突变型ADAM15结肠癌HCT-8细胞系细胞功能实验,进而完成野生型/突变型ADAM15细胞系的转录组测序分析。结果采用全外显子组测序在预后不良的患者中检出ADAM15基因SNV rs6427128,提示可能影响结肠癌的进展。细胞功能实验显示,rs6427128损失了ADAM15所具有的上调细胞侵袭能力的作用,而使细胞黏附能力增加约26%。转录组测序分析显示,与野生型相比,rs6427128过表达细胞的差异表达基因富集在细胞黏附、DNA的复制与转录、炎症反应等多种细胞生物学过程,提示rs6427128变异可能通过促进肿瘤细胞的黏附参与调节肿瘤微环境的重塑,从而影响结肠癌的进展。结论结合适当的细胞模型和较为精细的分析方法,深入解析临床样本中所检出的一些高频非同义潜在功能性变异体,可以为肿瘤相关基因的结构功能关系提供有益的提示性实验室数据。 展开更多
关键词 结肠癌 ADAM15基因 细胞黏附 单核苷酸变异 肿瘤微环境
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Polymorphism of p16INK4a gene and rare mutation of p15INK4b gene exon2 in primary hepatocarcinoma 被引量:30
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作者 Yang Qin Bo Li Yong Shu Tan Zhi Lin Sun Feng Qiong Zuo Ze Fang Sun Institute of Biochemistry and Molecular Biology,West China University of Medical Sciences,Chengdu 610041,Sichuan Province,China Department of General Surgery,The First Affiliated Hospital,West China University of Medical Sciences,Chengdu 610041,Sichuan Province,China Department of Pathology,The First Affiliated Hospital,West China University of Medical Sciences,Chengdu 610041,Sichuan Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2000年第3期411-414,共4页
INTRODUCTION Hepatocellular carcinoma(HCC)is the mostcommon cause of death from cancer in China.Themechanisms of hepatocarcinogenesis are not yetknown clearly,p16INK4a gene,the multiple tumorsuppressor gene 1(MTS1),en... INTRODUCTION Hepatocellular carcinoma(HCC)is the mostcommon cause of death from cancer in China.Themechanisms of hepatocarcinogenesis are not yetknown clearly,p16INK4a gene,the multiple tumorsuppressor gene 1(MTS1),encodes P16 protein,which acts as an inhibitor by binding directly toCDK4 and CDK6 and preventing its association 展开更多
关键词 P16INK4A gene P15INK4B gene POLYMORPHISM MUTATION HEPATOCARCINOMA
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ANTITUMOR EFFECTS OF HUMAN IL-15 GENE MODIFIED LUNG CANCER CELL LINE 被引量:2
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作者 沈永泉 崔莲仙 +2 位作者 何维 薛莉 巴德年 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 1997年第4期8-12,共5页
Human IL15 cDNA fragment, which contains all codons encoding the human IL15 mature protein and signal peptide was transducted into the human lung squmouse cancer cells(PG cells) and murine lung adenocarcinoma cells(LA... Human IL15 cDNA fragment, which contains all codons encoding the human IL15 mature protein and signal peptide was transducted into the human lung squmouse cancer cells(PG cells) and murine lung adenocarcinoma cells(LA795 cell lines). Two IL15 highly expressed cell clones PG1 and LA795A were used to inoculate the nude mice and the T739 syngeneic mice respectively. PG1 cell express higher level of class ⅠMHC molecule on their surface than PG cells. It was shown that the modified LA795A tumor cells grew slowly in T739 mice and induced high levels of CTL/NK/LAK activity in vivo as well, compared with the case of inoculation with LA795 or LA795neo. No significant difference in the tumor growth was observed in groups of the nude mice inoculated by PG1, PG and PGneo cells respectively, except the gene modified cells could not show the lung metastasis of tumors. The supernatants derived from the LA795A cell culture could promote CTL/NK/LAK activity from the whole splenocytes and the CD4/CD8deleted splenic cells in vitro. The results indicated that the IL15 gene transfected tumor cells play important roles in the process of antitumor or antitumor metastasis. 展开更多
关键词 INTERLEUKIN-15 gene therapy TUMOR gene transduction CYTOTOXICITY
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Expression Analysis of HMW-GS 1Bx14 and 1By15 in Wheat Varieties and Transgenic Research of 1By15 Gene 被引量:1
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作者 XU Tao ZHANG Xue-yong DONG Yu-shen 《Agricultural Sciences in China》 CAS CSCD 2006年第10期725-735,共11页
High-molecular-weight glutenin subunits (HMW-GSs), one class of seed storage proteins in wheat, play an important role in determining bread-making quality of flour. More and more proves support that HMW-GS- 1Bx 14 a... High-molecular-weight glutenin subunits (HMW-GSs), one class of seed storage proteins in wheat, play an important role in determining bread-making quality of flour. More and more proves support that HMW-GS- 1Bx 14 and - 1Bx 15 subunits are strongly positively associated with good bread-making and excellent noodle-making quality. The two subunits are encoded by two genes, Glu-1Bx14 and Glu-1Bx15, which are tightly linked and located on the 1BL. Protein assay by SDS-PAGE indicated that the expression of Glu-1Bx14 gene was always much stronger than that of Glu-1By15 in the same variety. But, variation of expression level for Glu-1By15 gene existed among varieties, such as in Xiaoyan 54, Xiaoyan 6, Yanzhan 1 and Shanyou 225. We also investigated the transcription difference of Glu-1By14 and Glu-1By15 genes in Xiaoyan 54 and Shanyou 225 by semi-quantitative RT-PCR method. The Glu-1By14 always transcripts much more than the Glu-1By15. This was basically consistent with the translation difference between the two genes. Promoters of 1Bx14, 1By15, 1By8, 1Dx2 and 1Dy12 were cloned from Xiaoyan 54, Chinese Spring and Aegilops tauschii. Sequence analysis indicated that the HMW-GS genes had high homology at their promoter regions. However, significant difference existed between sequence of 1Bx14 promoter and those of other HMW-GS genes. The transient expression experiment showed that the promoter of 1By15 has lower activity than that of 1Bx14, which was consistent with their transcription level of the two genes in varieties. In addition, transient expression of the gus driven by the promoter (P2) of HMW-GS 1Dx2 gene was higher than by other HMW-GS promoters. Therefore, we constructed 1By15 gene expression vector driven by the 1Dx2 promoter, and transformed the 1By15 gene into wheat commercial variety, Jimai 20 by pollen tube method. Of 45 independent transgenic lines identified by PCR, 3 were confirmed to contain the HMW-GS 1By15 gene via Southern hybridization. The delivered 1By15 gene expressed the expected HMW-GS protein in the seeds of transgenic plants. 展开更多
关键词 HMW-GS 1By15 gene transient expression assay TRANSGENIC
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白细胞介素15基因脂质体的构建及其抗肿瘤效果评价 被引量:1
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作者 江芷萱 宋宝慧 +3 位作者 常爽 张皓淞 胡海洋 陈大为 《沈阳药科大学学报》 CAS CSCD 北大核心 2023年第3期267-273,280,共8页
目的制备DSPE-PEG2000-FA修饰的阳离子脂质体(FPCL),并与白细胞介素15质粒DNA(interleukin-15 plasmid DNA,IL-15 pcDNA,pIL-15)形成FPCL/pIL-15基因递送系统,并进行抗肿瘤效果评价。方法通过薄膜分散法制备FPCL,并且利用正负电荷吸引... 目的制备DSPE-PEG2000-FA修饰的阳离子脂质体(FPCL),并与白细胞介素15质粒DNA(interleukin-15 plasmid DNA,IL-15 pcDNA,pIL-15)形成FPCL/pIL-15基因递送系统,并进行抗肿瘤效果评价。方法通过薄膜分散法制备FPCL,并且利用正负电荷吸引的原理,制备FPCL/pIL-15复合物。运用动态光散射法测定复合物粒径,利用琼脂糖凝胶电泳考察复合物稳定性,采用荧光分光光度法测定其包封率,以Balb/c小鼠为模型,考察制剂的抗肿瘤效果。结果成功制备了FPCL/pIL-15阳离子脂质体基因复合物,FPCL/pDNA复合物在质量比w_((DOTAP))∶w_((pDNA))=5∶1时粒径达到200 nm以下;抵御阴离子置换实验表明FPCL包载pDNA的稳定性良好,药效学实验表明FPCL/pIL-15复合物抗肿瘤效果显著。结论FPCL作为基因递送载体对基因药物具有很好的保护性,FPCL/pIL-15复合物能够有效诱导NK-细胞增殖与激活,抑瘤效果明显,在肿瘤免疫治疗领域有很大的研究价值。 展开更多
关键词 白细胞介素15 阳离子脂质体 基因递送 肿瘤免疫治疗 药效学
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Correlation Analysis of New Soybean[Glycine max(L.)Merr]Gene Gm15G117700 with Oleic Acid 被引量:1
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作者 Shuo Qu Yaolei Jiao +1 位作者 Lamboro Abraham Piwu Wang 《Phyton-International Journal of Experimental Botany》 SCIE 2021年第4期1177-1192,共16页
The fatty acid dehydrogenase gene plays an important role in regulating the oleic acid content in soybean.Genome-wide association study screened out soybean oleic acid related gene Gm15G117700.A fragment size of 693bp... The fatty acid dehydrogenase gene plays an important role in regulating the oleic acid content in soybean.Genome-wide association study screened out soybean oleic acid related gene Gm15G117700.A fragment size of 693bp was obtained by PCR amplification of the gene and,it was connected by seamless cloning technology to the pMD18T cloning vector.Based on the gene sequence cloned,bioinformatic analysis of gene protein was performed.The overexpression vector of Gm15G117700 and the CRISPR/Cas9 gene editing vector were constructed.The positive plants were obtained by Agrobacterium-mediated transformation of soybean cotyledon nodes and T2 plants were identified by conventional PCR,QT-PCR and Southern blot hybridization.10 copies of high and low oleic acid seeds were selected for QT-PCR to identify the expression content of Gm15G117700 gene in different soybeans,and finally near-infrared spectroscopy analyzer was used to identify the oleic acid quality of soybeans.T2 RT-PCR identification showed that overexpression was reduced by 3.94%,and gene editing was increased by 3.49%.It is determined that the Gm15G117700 gene may belong to a regulatory gene,a minor gene that can promote the conversion to linoleic acid content in soybean oleic acid synthesis.The gene cloning and its functional verification was not reported yet.This is the first report by PCR amplification of soybean Gm15G117700 genes and gene expression vector.Improving the content of oleic acid in soybean lay a foundation for researchers.Therefore;this study clearly identified the function of soybean Gm15G117700 gene and its role played in oleic acid synthesis and metabolism. 展开更多
关键词 SOYBEAN Gm15G117700 genes CLONING expression vector
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Advances on genetic and genomic studies of ALV resistance 被引量:1
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作者 Guodong Mo Ping Wei +2 位作者 Bowen Hu Qinghua Nie Xiquan Zhang 《Journal of Animal Science and Biotechnology》 SCIE CAS CSCD 2023年第1期1-14,共14页
Avian leukosis(AL)is a general term for a variety of neoplastic diseases in avian caused by avian leukosis virus(ALV).No vaccine or drug is currently available for the disease.Therefore,the disease can result in sever... Avian leukosis(AL)is a general term for a variety of neoplastic diseases in avian caused by avian leukosis virus(ALV).No vaccine or drug is currently available for the disease.Therefore,the disease can result in severe economic losses in poultry flocks.Increasing the resistance of poultry to ALV may be one effective strategy.In this review,we provide an overview of the roles of genes associated with ALV infection in the poultry genome,including endogenous retroviruses,virus receptors,interferon-stimulated genes,and other immune-related genes.Furthermore,some methods and techniques that can improve ALV resistance in poultry are discussed.The objectives are willing to provide some valuable references for disease resistance breeding in poultry. 展开更多
关键词 Avian leukosis Endogenous retrovirus gene editing IMMUNITY interferon-stimulated genes Receptor Resistant breeding
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Bone Morphogenetic Protein 15 as a Candidate Gene for Prolificacy of Jining Grey Goat
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作者 FENG Tao ZHAO You-zhang +3 位作者 CHU Ming-xing DI Ran ZHANG Ying-jie FANG Li 《畜牧兽医学报》 CAS CSCD 北大核心 2009年第S1期1-7,共7页
On the basis of the ovine bone morphogenetic protein 15(BMP15)gene,two pairs of primers(PI and P2)were designed to amplify exons 1 and 2 of the BMP15 gene in five randomly selected does of both Angora and Jining Grey ... On the basis of the ovine bone morphogenetic protein 15(BMP15)gene,two pairs of primers(PI and P2)were designed to amplify exons 1 and 2 of the BMP15 gene in five randomly selected does of both Angora and Jining Grey goats.The sequences of BMP15 exon 1(P1 amplification)of Angora and Jining Grey goats were identical.There was a 3-nucleotide(CTT)insertion in positions 268 to 270 of goat BMP 15 exon1 compared with that of sheep(GenBank accession number AF236078),which caused a leucine insertion in the 12th position of amino acid sequence.Sequence length of goat BMP 15 exon 2(P2 amplification)was identical with that of sheep(AF236079),but there were seven nucleotide and four amino acid changes between goat and sheep.The nucleotide in the 963rd position of BMP15 exon 2 was A for Angora goat and sheep,and G for Jining Grey goat.Based on this A963G mutation,primer pair P3 was designed to detect single nucleotide polymorphism of BMP15 exon 2 in breeds of high prolificacy(Jining Grey),moderate prolificacy(Boer)and low prolificacy(Angora and Inner Mongolia Cashmere)by polymerase chain reactionsingle strand conformation polymorphism(PCR-SSCP).Three genotypes(AA,AG and GG)were detected in Jining Grey goats,two genotypes(AG and GG)in Boer,and only the AA genotype in Angora and Inner Mongolia Cashmere goats.Sequencing revealed one mutation(A963G)in genotype GG compared with genotype AA,and this mutation resulted in an amino acid change of serine→glycine(S300G).In Jining Grey goats,frequencies of AA,AG and GG genotypes were 0.008,0.059 and 0.933,respectively.Genotypic distributions of the BMP 15 gene were significantly different(P<0.05 or P<0.001)between Jining Grey and Boer,Angora,and Inner Mongolia Cashmere goats.In Jining Grey goats,the does with the GG genotype had 0.71(P<0.05)or 1.57(P<0.05)additional kids than did those with AG or AA genotypes,and does with the AG genotype had 0.86(P<0.05)more kids than did those with the AA genotype.These results tentatively indicate that the BMP15 gene is either a major gene that affects prolificacy in Jining Grey goats,or may be a molecular marker in close linkage with such a gene. 展开更多
关键词 GOAT PROLIFICACY BONE morphogenetic protein 15 gene PCR-SSCP
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干扰素刺激基因15抗病毒感染的分子机制
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作者 唐井玉 杜汉宇 +6 位作者 贾楠楠 汤傲星 刘春草 朱杰 孟春春 李传峰 刘光清 《中国动物传染病学报》 CAS 北大核心 2023年第6期170-176,共7页
干扰素刺激基因15(ISG15)是由病原微生物或干扰素诱导产生的一种大小为15 kDa的泛素样蛋白。在干扰素诱导的数百个干扰素刺激基因中,ISG15是诱导最强烈、最快的ISG蛋白之一。研究表明,ISG15对多种病毒具有抗病毒作用。此外,ISG15在调节... 干扰素刺激基因15(ISG15)是由病原微生物或干扰素诱导产生的一种大小为15 kDa的泛素样蛋白。在干扰素诱导的数百个干扰素刺激基因中,ISG15是诱导最强烈、最快的ISG蛋白之一。研究表明,ISG15对多种病毒具有抗病毒作用。此外,ISG15在调节宿主损伤、DNA修复,调节信号通路及抗原递呈中也发挥着重要的作用。文章介绍了ISG15的概况,并阐述了近年来ISG15在抗病毒、免疫调节和调节宿主信号通路过程中的作用。 展开更多
关键词 干扰素刺激基因15 抗病毒作用 免疫调节
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Detection of the FecX^R Mutation of BMP15 Gene in Sheep and Goats
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作者 DI Ran FENG Tao +2 位作者 CHU Ming-xing ZHANG Ying-jie FANG Li 《畜牧兽医学报》 CAS CSCD 北大核心 2010年第S1期17-20,共4页
PCR-SSCP was used to detect mutations of bone morphogenetic protein 15(BMP15) gene in both high prolificacy(Small Tail Han sheep,Hu sheep,Jining Grey goat and Boer goat) and low prolificacy breeds(Dorset sheep,Texel s... PCR-SSCP was used to detect mutations of bone morphogenetic protein 15(BMP15) gene in both high prolificacy(Small Tail Han sheep,Hu sheep,Jining Grey goat and Boer goat) and low prolificacy breeds(Dorset sheep,Texel sheep,Inner Mongolia Cashmere goat and Angora goat).Both the nucleotide sequences and the amino acid sequences were compared in amplification fragments of both Small Tail Han sheep and Jining Grey goat.The results indicated that none of the four sheep and the four goat breeds carried the same FecX<sup>R</sup> mutation of the BMP15 gene as do Rasa Aragonesa sheep.The nucleotide sequence of Small Tail Han sheep was completely identical with that of the sheep BMP15 sequence(GenBank AF236079,NM<sub>0</sub>01114767).Three base substitutions(T529G,C530G and T576C) and two amino acid changes(V155G and S171P) were found in Jining Grey goat compared with Small Tail Han sheep.The FecX<sup>R</sup> mutation of the BMP15 gene had no significant effect on high prolificacy of Small Tail Han sheep, Hu sheep,Jining Grey goat and Boer goat. 展开更多
关键词 SHEEP GOAT PROLIFICACY BMP15 gene FecX^R mutation PCR-SSCP
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罕见病研究:GPAA1基因突变导致糖基磷脂酰肌醇生物合成缺陷15型
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作者 陈秋蓉 张朕杰 +2 位作者 卢一岫 袁孙碧歆 李冀 《中国当代儿科杂志》 CAS CSCD 北大核心 2023年第12期1276-1281,共6页
6岁男性患儿,因发育迟缓6年,反复发热、抽搐5年就诊。患儿3月龄时发现精神运动发育落后,1岁起出现反复发热、抽搐,伴间断口腔溃疡及扁桃体化脓,发热期间查血白细胞计数、C反应蛋白、红细胞沉降率升高,热退后正常,脑电图提示癫痫,基因检... 6岁男性患儿,因发育迟缓6年,反复发热、抽搐5年就诊。患儿3月龄时发现精神运动发育落后,1岁起出现反复发热、抽搐,伴间断口腔溃疡及扁桃体化脓,发热期间查血白细胞计数、C反应蛋白、红细胞沉降率升高,热退后正常,脑电图提示癫痫,基因检测提示GPAA1基因存在复合杂合突变。最终该患儿诊断为糖基磷脂酰肌醇生物合成缺陷15型(glycosylphosphatidylinositol biosynthesis deficiency 15,GPIBD15)、周期性发热。该患儿抗癫痫效果不佳,糖皮质激素治疗对发热有效。该文报道了中国首例GPAA1基因突变导致GPIBD15患儿,对该病基因、临床特点、诊疗等进行归纳总结,为该病的早期诊断、治疗提供参考依据。 展开更多
关键词 糖基磷脂酰肌醇生物合成缺陷15 GPAA1基因 癫痫 周期性发热 儿童
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GDF-15基因多态性、膳食与中心性肥胖的相关性研究
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作者 梁玉坚 王雪秀 +3 位作者 覃丽冬 蔡建升 覃健 张志勇 《华夏医学》 CAS 2023年第3期1-7,共7页
目的:探究GDF-15基因多态性与膳食因素的交互作用及其与中心性肥胖的关联。方法:采用Sequenom MassArray系统对GDF-15基因多态性分型。使用食物频率问卷(FFQ)法调查膳食摄入情况。应用Logistic回归分析SNP位点、膳食因素及两者间交互作... 目的:探究GDF-15基因多态性与膳食因素的交互作用及其与中心性肥胖的关联。方法:采用Sequenom MassArray系统对GDF-15基因多态性分型。使用食物频率问卷(FFQ)法调查膳食摄入情况。应用Logistic回归分析SNP位点、膳食因素及两者间交互作用与中心性肥胖的关联。结果:Logistics回归分析显示,携带GDF-15基因rs1059519CG基因型人群患中心性肥胖的风险高于携带GDF-15基因rs1059519GG基因型人群,OR为1.986(95%CI:1.011~3.900,P<0.05)。饱和脂肪酸、多不饱和脂肪酸的中等摄入量与中心性肥胖的风险相关,OR分别为1.983(95%CI:1.139~3.451,P<0.05)和2.146(95%CI:1.238~3.719,P<0.05)。rs1059519CG基因型与饱和脂肪酸、多不饱和脂肪酸中等摄入量的交互作用会增加中心性肥胖的风险,OR分别为2.737(95%CI:1.241~4.536,P<0.05)和2.852(95%CI:1.540~5.281,P<0.05)。结论:GDF-15基因多态性、饱和脂肪酸、多不饱和脂肪酸摄入及两者间交互作用与中心性肥胖相关。 展开更多
关键词 GDF-15基因多态性 中心性肥胖 膳食脂肪酸
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乳腺癌组织中泛素特异性蛋白酶18/干扰素刺激基因15表达水平及其临床意义 被引量:1
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作者 蔡冰 尹香利 刘静 《安徽医药》 CAS 2023年第7期1449-1454,I0003,共7页
目的探究泛素特异性蛋白酶18(USP18)、干扰素刺激基因15(ISG15)在乳腺癌组织中的表达水平及临床意义。方法采用Ualcan数据库分析USP18、ISG15在正常乳腺组织和乳腺癌组织中的表达情况及二者与乳腺癌预后的关系;选取渭南市中心医院2012年... 目的探究泛素特异性蛋白酶18(USP18)、干扰素刺激基因15(ISG15)在乳腺癌组织中的表达水平及临床意义。方法采用Ualcan数据库分析USP18、ISG15在正常乳腺组织和乳腺癌组织中的表达情况及二者与乳腺癌预后的关系;选取渭南市中心医院2012年5月至2015年7月诊治的99例乳腺癌病人癌组织、癌旁正常组织进行研究。分别检测USP18、ISG15mRNA及其蛋白表达情况;分析乳腺癌组织USP18、ISG15表达水平与病人临床病理特征、预后的关系;Cox回归分析乳腺癌病人预后的影响因素。结果Ualcan数据库中乳腺癌组织USP18为18.40±4.17、ISG15 mRNA表达水平为168.56±43.95高于正常乳腺组织(10.00±3.14、30.76±8.23)(P<0.05);乳腺癌组织USP18、ISG15 mRNA表达水平高低与乳腺癌预后无明显相关性。乳腺癌组织USP18为1.67±0.53、ISG15 mRNA为1.86±0.61及蛋白阳性表达率均高于癌旁正常组织(1.03±0.34、0.99±0.33)(P<0.05);乳腺癌组织USP18、ISG15表达水平均与淋巴结转移、乳腺癌分子亚型、肿瘤分化程度、TNM分期相关(P<0.05);乳腺癌病人癌组织中USP18表达水平与ISG15呈正相关(P<0.05);USP18阳性组、ISG15阳性组乳腺癌病人术后60个月生存率均低于USP18阴性组、ISG15阴性组(P<0.05);淋巴结转移、TNM分期、USP18、ISG15均是影响乳腺癌病人不良预后的独立危险因素(P<0.05)。结论乳腺癌病人癌组织USP18、ISG15表达水平均较高,两者均与乳腺癌病人预后关系密切,检测乳腺癌组织USP18、ISG15水平有助于评估乳腺癌病人预后。 展开更多
关键词 乳腺肿瘤 干扰素刺激基因15 泛素特异性蛋白酶18 预后
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Analysis of differentially expressed genes in Verruca vulgaris vs.adjacent normal skin by RNA-sequencing
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作者 QINGQING GUO JIAYUE QI +4 位作者 XIAOQIANG LIANG ZIGANG ZHAO JIA BAI FANG XIE CHENGXIN LI 《BIOCELL》 SCIE 2023年第11期2435-2443,共9页
Introduction:Verruca vulgaris is one of the most common low-risk HPV infections and is characterized by excessive proliferation of keratinocytes.Currently,very little genetic information is available regarding verruca... Introduction:Verruca vulgaris is one of the most common low-risk HPV infections and is characterized by excessive proliferation of keratinocytes.Currently,very little genetic information is available regarding verruca vulgaris in the Chinese population.This study aimed to obtain comprehensive transcript information of verruca vulgaris by RNA sequencing.Methods:High-throughput sequencing was performed on three fresh verruca vulgaris samples and adjacent normal skin on the Illumina sequencing platform.The transcriptomes were analyzed using bioinformatics and the differentially expressed genes(DEGs)were verified by immunohistochemistry.Verruca vulgaris exhibited a unique molecular signature.Results:In total,1,643 DEGs were identified in verruca vulgaris compared to normal skin.The functions of the DEGs were studies by Gene Ontology(GO)enrichment,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,DEGs Reactome analysis,disease annotation function,and STRING protein-protein interaction(PPI)network analysis.The results revealed 595 GO terms associated with the cell cycle,signal transduction,immune system,signaling molecules,and interaction.The Reactome analysis revealed enrichment in reversible hydration of carbon dioxide and BMP signaling,while the disease annotation function revealed that the enriched DEGs are involved in keratosis disorders.The STRING PPI network showed that the edges with the highest density mainly included the 2′-5′oligoadenylate synthase(OAS)family-related proteins.Furthermore,the M-code analysis found ISG15,IRF7,and OASL were scored as significant modules and their high expression compared to the control was verified by immunohistochemistry.Conclusion:These findings contribute to the genetic information of verruca vulgaris in the Chinese population,revealing that interferon-stimulated genes may play essential roles in verruca vulgaris. 展开更多
关键词 Differentially expressed genes RNA-SEQ TRANSCRIPTOME Verruca vulgaris interferon-stimulated genes
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lncRNA SNHG15通过调控miR-942-5p表达减轻Aβ25-35诱导的PC12细胞氧化应激和细胞凋亡的研究
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作者 张文娟 戴婷丽 李沛 《国际检验医学杂志》 CAS 2023年第23期2880-2885,共6页
目的探讨长链非编码RNA(lncRNA)小核仁RNA宿主基因15(SNHG15)对β淀粉样蛋白_(25-35)(Aβ_(25-35))诱导的PC12细胞氧化应激和细胞凋亡的影响及分子机制。方法将PC12细胞分为Con组、Aβ_(25-35)组、Aβ_(25-35)+si-SNHG15组、Aβ_(25-35)... 目的探讨长链非编码RNA(lncRNA)小核仁RNA宿主基因15(SNHG15)对β淀粉样蛋白_(25-35)(Aβ_(25-35))诱导的PC12细胞氧化应激和细胞凋亡的影响及分子机制。方法将PC12细胞分为Con组、Aβ_(25-35)组、Aβ_(25-35)+si-SNHG15组、Aβ_(25-35)+si-NC组、Aβ_(25-35)+微小RNA(miR)-942-5p组、Aβ_(25-35)+miR-NC组、Aβ_(25-35)+si-SNHG15+anti-miR-942-5p组、Aβ_(25-35)+si-SNHG15+anti-miR-NC组;实时荧光定量PCR(qRT-PCR)检测lncRNA SNHG15和miR-942-5p表达水平;酶联免疫吸附试验检测细胞超氧化物歧化酶(SOD)活性和丙二醛(MDA)表达水平;蛋白质印迹法检测蛋白表达;流式细胞术检测细胞凋亡;双荧光素酶报告实验检测lncRNA SNHG15和miR-942-5p靶向关系。结果Aβ_(25-35)诱导的PC12细胞中lncRNA SNHG15、MDA、Bax表达水平及细胞凋亡率升高,SOD活性、miR-942-5p、Bcl-2表达水平降低(P<0.05)。抑制lncRNA SNHG15表达或过表达miR-942-5p后,SOD活性、Bcl-2表达水平升高,细胞凋亡率、MDA、Bax表达水平降低(P<0.05)。lncRNA SNHG15靶向调控miR-942-5p。下调miR-942-5p表达逆转了抑制lncRNA SNHG15表达对Aβ_(25-35)诱导的PC12细胞氧化应激和凋亡的作用。结论抑制lncRNA SNHG15表达通过上调miR-942-5p表达减轻Aβ_(25-35)诱导的PC12细胞氧化应激和细胞凋亡。 展开更多
关键词 长链非编码RNA小核仁RNA宿主基因15 微小RNA-942-5p 氧化应激 细胞凋亡
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Anti-aging Effects of Alu Antisense RNA on Human Fibroblast Senescence Through the MEK-ERK Pathway Mediated by KIF15
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作者 Ning JI Chong-guang WU +7 位作者 Xiao-die WANG Zhi-xue SONG Pei-yuan WU Xin LIU Xu FENG Xiang-mei ZHANG Xiu-fang WANG Zhan-jun LV 《Current Medical Science》 SCIE CAS 2023年第1期35-47,共13页
Objective:To investigate whether human short interspersed nuclear element antisense RNA(Alu antisense RNA;Alu asRNA)could delay human fibroblast senescence and explore the underlying mechanisms.Methods:We transfected ... Objective:To investigate whether human short interspersed nuclear element antisense RNA(Alu antisense RNA;Alu asRNA)could delay human fibroblast senescence and explore the underlying mechanisms.Methods:We transfected Alu asRNA into senescent human fibroblasts and used cell counting kit-8(CCK-8),reactive oxygen species(ROS),and senescence-associated beta-galactosidase(SA-β-gal)staining methods to analyze the anti-aging effects of Alu asRNA on the fibroblasts.We also used an RNA-sequencing(RNA-seq)method to investigate the Alu asRNA-specific mechanisms of anti-aging.We examined the effects of KIF15 on the anti-aging role induced by Alu asRNA.We also investigated the mechanisms underlying a KIF15-induced proliferation of senescent human fibroblasts.Results:The CCK-8,ROS and SA-β-gal results showed that Alu asRNA could delay fibroblast aging.RNA-seq showed 183 differentially expressed genes(DEGs)in Alu asRNA transfected fibroblasts compared with fibroblasts transfected with the calcium phosphate transfection(CPT)reagent.The KEGG analysis showed that the cell cycle pathway was significantly enriched in the DEGs in fibroblasts transfected with Alu asRNA compared with fibroblasts transfected with the CPT reagent.Notably,Alu asRNA promoted the KIF15 expression and activated the MEK-ERK signaling pathway.Conclusion:Our results suggest that Alu asRNA could promote senescent fibroblast proliferation via activation of the KIF15-mediated MEK-ERK signaling pathway. 展开更多
关键词 senescent fibroblast cell proliferation Alu antisense RNA KIF15 gene expression MEK-ERK signaling pathway cell cycle
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LncRNA SNHG15/miR-123/PAK5轴通过自噬对胃癌细胞活性及血管生成的影响
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作者 刘杰 刘树青 +2 位作者 梁凤 王蕾 刘永健 《解剖学研究》 CAS 2023年第3期242-250,共9页
目的探讨lncRNA SNHG15/miR-123/PAK5轴通过调节自噬对胃癌细胞活性及血管生成的机制研究。方法将胃癌SCG-823细胞分为Control组、si-NC组、si-SNHG15组、miR-NC组、miR-123组、pcDNAPAK5组。RT-PCR检测细胞中SNHG15和miR-123表达;MTT... 目的探讨lncRNA SNHG15/miR-123/PAK5轴通过调节自噬对胃癌细胞活性及血管生成的机制研究。方法将胃癌SCG-823细胞分为Control组、si-NC组、si-SNHG15组、miR-NC组、miR-123组、pcDNAPAK5组。RT-PCR检测细胞中SNHG15和miR-123表达;MTT法检测细胞增殖能力;Transwell小室法检测细胞侵袭能力;流式细胞仪检测细胞凋亡能力;Matrigel体外成管实验检测细胞血管生成能力;蛋白质印迹检测细胞中PAK5、VEGF、LC3、Beclin1、p62蛋白表达;双荧光素酶报告基因检验SNHG15和miR-123、miR-123和PAK5的靶向关系。结果与人胃黏膜细胞系GES-1相比,人胃癌细胞系中ASG、MKN-45、SCG-823细胞中SNHG15表达明显升高,miR-123表达明显降低(P<0.05);且SCG-823细胞中SNHG15表达明显高于ASG、MKN-45细胞,miR-123表达明显低于ASG、MKN-45细胞(P<0.05)。si-SNHG15组细胞增殖、侵袭及形成小管数量均明显低于Control组,细胞凋亡率高于Control组(P<0.05);si-SNHG15组细胞中VEGF、PAK5、p62蛋白表达明显低于Control组,LC3Ⅱ/LC3Ι比值及Beclin1蛋白表达明显高于Control组(P<0.05)。与miR-NC组相比,miR-123组细胞增殖、侵袭及形成小管数量均明显降低,细胞凋亡率明显增加(P<0.05);miR-123组细胞中VEGF、PAK5、p62蛋白表达明显低于miR-NC组组,LC3Ⅱ/LC3Ι比值及Beclin1蛋白表达明显高于miRNC组(P<0.05)。通过向细胞中分别转染野生型SNHG15(SNHG15-WT)、PAK5(SNHG15-WT)时,miR-123组荧光素酶活性均明显低于miR-NC组(P<0.05)。与si-SNHG15组相比,pcDNA-PAK5组细胞细胞增殖、侵袭及形成小管数量均明显升高,细胞凋亡率明显降低(P<0.05);pcDNA-PAK5组细胞中VEGF、PAK5、p62蛋白表达明显高于si-SNHG15组,LC3Ⅱ/LC3Ι比值及Beclin1蛋白表达明显低于si-SNHG15组(P<0.05)。结论lncRNA SNHG15可靶向miR-123/PAK5轴抑制胃癌细胞增殖、侵袭和血管生成,促进胃癌细胞凋亡和自噬,进而为调控自噬途径治疗胃癌提供新的思路。 展开更多
关键词 人胃癌细胞株 血管生成 自噬 lncRNA SNHG15/miR-123/PAK5轴 小核仁RNA宿主基因15 增殖
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