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
In this study, using Liangshan semi-fine wool sheep as an experimental material, full-length CDS sequence of IGFBP-1 gene was cloned with RT-PCR method and analyzed with bioinformatics methods. The results showed that...In this study, using Liangshan semi-fine wool sheep as an experimental material, full-length CDS sequence of IGFBP-1 gene was cloned with RT-PCR method and analyzed with bioinformatics methods. The results showed that the full-length CDS sequence of IGFBP-1 gene in Liangshan semi-fine wool sheep was 792 bp in length, encoding 263 amino acids. The CDS sequence shared 97%, 76% and 74% homology with bovine, human and rat, respectively; the amino acid sequence shared 97% , 69% and 71%, respectively. The GenBank accession number was FJ589639.1. The amino acid molecular weight of IGFBP-1 was 27.8 kD, and the theoretical isoelectric point （pl） was 5.99. The result of phylogenetic analysis showed that IGFBP-1 gene in Liangshan semi-fine wool sheep exhibited close phylogenetic relationships with bovine, goat and other mammals, and distant phylogenetic relationships with chicken and fish. IGFBP-1 gene had obvious hy- drophobic and hydrophilic regions, harboring one signal peptide, one transmembrane region, 16 phosphorylation sites, six N-glycesylation sites and eight O-glycosy- lation sites. The result of secondary structure analysis showed that the random coil, a-helix and 13-sheet regions accounted for 61.98%, 24.33% and 13.69%, re- spectively. The result of tertiary structure analysis showed that IGFBP-1 harbors an IGFBP_N domain and a thyrnglobulin type-1 domain. This study laid a solid foundation for further investigating the function of IGFBP-1 gene in sheep.展开更多
试验旨在分析牦牛跨膜附睾蛋白1(transmembrane epididymal protein 1,TEDDM1)基因的分子特征及其在卵泡发育中的时序表达特性。研究分别收集发情期牦牛大卵泡(≥8 mm)、中卵泡(=5~7 mm)和小卵泡(≤3 mm)中的卵丘-卵母细胞复合体(...试验旨在分析牦牛跨膜附睾蛋白1(transmembrane epididymal protein 1,TEDDM1)基因的分子特征及其在卵泡发育中的时序表达特性。研究分别收集发情期牦牛大卵泡(≥8 mm)、中卵泡(=5~7 mm)和小卵泡(≤3 mm)中的卵丘-卵母细胞复合体(COCs);提取总RNA并反转录,对TEDDM1基因CDS区全序列进行克隆测序,用生物信息学软件分析其编码蛋白分子特征;以牛GAPDH基因为内参,采用实时荧光定量PCR技术分析TEDDM1基因在卵泡发育不同时期的表达水平。结果表明,牦牛TEDDM1基因CDS区全长903 bp,共编码300个氨基酸,与野牦牛、牛、野牛、绵羊和水牛氨基酸序列有极高的相似性,在进化中较为保守;牦牛TEDDM1蛋白为非分泌型不稳定疏水蛋白,共存在7个螺旋结构,整条肽链7次横跨胞膜,属于典型的G蛋白偶联受体;其潜在的磷酸化位点共21个,其中酪氨酸、苏氨酸和丝氨酸磷酸化位点分别为1、3和17个,仅存在1个N-糖基化位点,无O-糖基化位点;存在未知功能结构域蛋白家族(domains of unknow function protein families,DUFs)716结构域,且该结构域涵盖多个跨膜区域;实时荧光定量PCR检测结果发现,中卵泡中TEDDM1基因表达水平显著高于大卵泡和小卵泡(P<0.05),而大卵泡与小卵泡的表达水平差异不显著(P>0.05)。本研究分析了牦牛TEDDM1基因序列及其在牦牛COCs中的表达特性,为进一步揭示该基因在牦牛卵泡发育过程中的调控作用奠定了理论基础。展开更多
Aim： To investigate the possible functions of human sperm membrane protein （hSMP-1） in the process of fertilization. Methods： A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was ...Aim： To investigate the possible functions of human sperm membrane protein （hSMP-1） in the process of fertilization. Methods： A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 （anti-rhSMP-1） antibodies. Sperm acrosome reaction and spermzona pellucida （ZP） binding assay were carried out in 10-week-old BALB/c mice. Results： Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 （P 〈 0.05）. Conclusion： The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.展开更多
The complete gene coding for human neutrophilactivating protein-1/interleukin-8 was synthesized using a semi-chemical semi-enzymatic method. The synthetic gene was then overexpressed in Escherichia coli under the temp...The complete gene coding for human neutrophilactivating protein-1/interleukin-8 was synthesized using a semi-chemical semi-enzymatic method. The synthetic gene was then overexpressed in Escherichia coli under the temperature-regulated control of the P_RP_L tandem promoters. As determined by SDS-PAGE and densitometry, the overexpressed protein comprised up to 18.5% and 10.9% of the total soluble protein in E. coli cells grown in shake flasks and in batch fermentation, respectively. The recombinant NAP-1/IL-8 was then purified to】95% homogeneity by gel filtration and cation exchange chromatography. The purified protein appeared as a single band on the SDS-PAGE gel and possessed potent chemotactic activity in the concentration of 【10 ng/ml, as assayed by the agarose plate method. An early skin reactivity was also observed when the pure NAP-1/IL-8 was injected subcutaneously into the rabbits. The N-terminal 36 amino acid sequence of the recombinant NAP1/IL-8 was determined using the Edman method and was展开更多
In a previous study,we found that long non-coding genes in Alzheimer’s disease(AD)are a result of endogenous gene disorders caused by the recruitment of microRNA(miRNA)and mRNA,and that miR-200a-3p and other represen...In a previous study,we found that long non-coding genes in Alzheimer’s disease(AD)are a result of endogenous gene disorders caused by the recruitment of microRNA(miRNA)and mRNA,and that miR-200a-3p and other representative miRNAs can mediate cognitive impairment and thus serve as new biomarkers for AD.In this study,we investigated the abnormal expression of miRNA and mRNA and the pathogenesis of AD at the epigenetic level.To this aim,we performed RNA sequencing and an integrative analysis of the cerebral cortex of the widely used amyloid precursor protein and presenilin-1 double transgenic mouse model of AD.Overall,129 mRNAs and 68 miRNAs were aberrantly expressed.Among these,eight down-regulated miRNAs and seven up-regulated miRNAs appeared as promising noninvasive biomarkers and therapeutic targets.The main enriched signaling pathways involved mitogen-activated kinase protein,phosphatidylinositol 3-kinase-protein kinase B,mechanistic target of rapamycin kinase,forkhead box O,and autophagy.An miRNA-mRNA network between dysregulated miRNAs and corresponding target genes connected with AD progression was also constructed.These miRNAs and mRNAs are potential biomarkers and therapeutic targets for new treatment strategies,early diagnosis,and prevention of AD.The present results provide a novel perspective on the role of miRNAs and mRNAs in AD.This study was approved by the Experimental Animal Care and Use Committee of Institute of Medicinal Biotechnology of Beijing,China(approval No.IMB-201909-D6)on September 6,2019.展开更多
c-Jun,the most extensively studied protein of the activator protein-1(AP-1)complex,is involved in numerous cell activities,such as proliferation,apoptosis,survival,tumorigenesis and tissue morphogenesis.Earlier studie...c-Jun,the most extensively studied protein of the activator protein-1(AP-1)complex,is involved in numerous cell activities,such as proliferation,apoptosis,survival,tumorigenesis and tissue morphogenesis.Earlier studies focused on the structure and function have led to the identification of c-Jun as a basic leucine zipper(bZIP)transcription factor that acts as homo-or heterodimer,binding to DNA and regulating gene transcription.Later on,it was shown that extracellular signals can induce post-translational modifications of c-Jun,resulting in altered transcriptional activity and target gene expression.More recent work has uncovered multiple layers of a complex regulatory scheme in which c-Jun is able to crosstalk,amplify and integrate different signals for tissue development and disease.One example of such scheme is the autocrine amplification loop,in which signal-induced AP-1 activates the c-Jun gene promoter,while increased c-Jun expression feedbacks to potentiate AP-1 activity.Another example of such scheme,based on recent characterization of gene knockout mice,is that c-Jun integrates signals of several developmental pathways,including EGFR-ERK,EGFR-RhoA-ROCK,and activin B-MAP3K1-JNK for embryonic eyelid closure.After more than two decades of extensive research,c-Jun remains at the center stage of a molecular network with mysterious functional properties,some of which are yet to be discovered.In this article,we will provide a brief historical overview of studies on c-Jun regulation and function,and use eyelid development as an example to illustrate the complexity of c-Jun crosstalking with signaling pathways.展开更多
文摘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.
基金Supported by Special Fund for Agro-scientific Research in the Public Interest from the Ministry of Agriculture(201003061)Key Project of Livestock and Poultry Breeding of Sichuan Province(01NG029-18)
文摘In this study, using Liangshan semi-fine wool sheep as an experimental material, full-length CDS sequence of IGFBP-1 gene was cloned with RT-PCR method and analyzed with bioinformatics methods. The results showed that the full-length CDS sequence of IGFBP-1 gene in Liangshan semi-fine wool sheep was 792 bp in length, encoding 263 amino acids. The CDS sequence shared 97%, 76% and 74% homology with bovine, human and rat, respectively; the amino acid sequence shared 97% , 69% and 71%, respectively. The GenBank accession number was FJ589639.1. The amino acid molecular weight of IGFBP-1 was 27.8 kD, and the theoretical isoelectric point （pl） was 5.99. The result of phylogenetic analysis showed that IGFBP-1 gene in Liangshan semi-fine wool sheep exhibited close phylogenetic relationships with bovine, goat and other mammals, and distant phylogenetic relationships with chicken and fish. IGFBP-1 gene had obvious hy- drophobic and hydrophilic regions, harboring one signal peptide, one transmembrane region, 16 phosphorylation sites, six N-glycesylation sites and eight O-glycosy- lation sites. The result of secondary structure analysis showed that the random coil, a-helix and 13-sheet regions accounted for 61.98%, 24.33% and 13.69%, re- spectively. The result of tertiary structure analysis showed that IGFBP-1 harbors an IGFBP_N domain and a thyrnglobulin type-1 domain. This study laid a solid foundation for further investigating the function of IGFBP-1 gene in sheep.
文摘试验旨在分析牦牛跨膜附睾蛋白1(transmembrane epididymal protein 1,TEDDM1)基因的分子特征及其在卵泡发育中的时序表达特性。研究分别收集发情期牦牛大卵泡(≥8 mm)、中卵泡(=5~7 mm)和小卵泡(≤3 mm)中的卵丘-卵母细胞复合体(COCs);提取总RNA并反转录,对TEDDM1基因CDS区全序列进行克隆测序,用生物信息学软件分析其编码蛋白分子特征;以牛GAPDH基因为内参,采用实时荧光定量PCR技术分析TEDDM1基因在卵泡发育不同时期的表达水平。结果表明,牦牛TEDDM1基因CDS区全长903 bp,共编码300个氨基酸,与野牦牛、牛、野牛、绵羊和水牛氨基酸序列有极高的相似性,在进化中较为保守;牦牛TEDDM1蛋白为非分泌型不稳定疏水蛋白,共存在7个螺旋结构,整条肽链7次横跨胞膜,属于典型的G蛋白偶联受体;其潜在的磷酸化位点共21个,其中酪氨酸、苏氨酸和丝氨酸磷酸化位点分别为1、3和17个,仅存在1个N-糖基化位点,无O-糖基化位点;存在未知功能结构域蛋白家族(domains of unknow function protein families,DUFs)716结构域,且该结构域涵盖多个跨膜区域;实时荧光定量PCR检测结果发现,中卵泡中TEDDM1基因表达水平显著高于大卵泡和小卵泡(P<0.05),而大卵泡与小卵泡的表达水平差异不显著(P>0.05)。本研究分析了牦牛TEDDM1基因序列及其在牦牛COCs中的表达特性,为进一步揭示该基因在牦牛卵泡发育过程中的调控作用奠定了理论基础。
文摘Aim： To investigate the possible functions of human sperm membrane protein （hSMP-1） in the process of fertilization. Methods： A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 （anti-rhSMP-1） antibodies. Sperm acrosome reaction and spermzona pellucida （ZP） binding assay were carried out in 10-week-old BALB/c mice. Results： Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 （P 〈 0.05）. Conclusion： The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.
文摘The complete gene coding for human neutrophilactivating protein-1/interleukin-8 was synthesized using a semi-chemical semi-enzymatic method. The synthetic gene was then overexpressed in Escherichia coli under the temperature-regulated control of the P_RP_L tandem promoters. As determined by SDS-PAGE and densitometry, the overexpressed protein comprised up to 18.5% and 10.9% of the total soluble protein in E. coli cells grown in shake flasks and in batch fermentation, respectively. The recombinant NAP-1/IL-8 was then purified to】95% homogeneity by gel filtration and cation exchange chromatography. The purified protein appeared as a single band on the SDS-PAGE gel and possessed potent chemotactic activity in the concentration of 【10 ng/ml, as assayed by the agarose plate method. An early skin reactivity was also observed when the pure NAP-1/IL-8 was injected subcutaneously into the rabbits. The N-terminal 36 amino acid sequence of the recombinant NAP1/IL-8 was determined using the Edman method and was
基金This study was supported by the National Natural Science Foundation of China(General Program),No.81673411the United Fund Project of National Natural Science Foundation of China,No.U1803281+1 种基金Young Medical Talents Award Project of Chinese Academy of Medical Sciences,No.2018RC350013Chinese Academy of Medical Sciences Innovation Project for Medical Science,No.2017-I2M-1-016(all to RL).
文摘In a previous study,we found that long non-coding genes in Alzheimer’s disease(AD)are a result of endogenous gene disorders caused by the recruitment of microRNA(miRNA)and mRNA,and that miR-200a-3p and other representative miRNAs can mediate cognitive impairment and thus serve as new biomarkers for AD.In this study,we investigated the abnormal expression of miRNA and mRNA and the pathogenesis of AD at the epigenetic level.To this aim,we performed RNA sequencing and an integrative analysis of the cerebral cortex of the widely used amyloid precursor protein and presenilin-1 double transgenic mouse model of AD.Overall,129 mRNAs and 68 miRNAs were aberrantly expressed.Among these,eight down-regulated miRNAs and seven up-regulated miRNAs appeared as promising noninvasive biomarkers and therapeutic targets.The main enriched signaling pathways involved mitogen-activated kinase protein,phosphatidylinositol 3-kinase-protein kinase B,mechanistic target of rapamycin kinase,forkhead box O,and autophagy.An miRNA-mRNA network between dysregulated miRNAs and corresponding target genes connected with AD progression was also constructed.These miRNAs and mRNAs are potential biomarkers and therapeutic targets for new treatment strategies,early diagnosis,and prevention of AD.The present results provide a novel perspective on the role of miRNAs and mRNAs in AD.This study was approved by the Experimental Animal Care and Use Committee of Institute of Medicinal Biotechnology of Beijing,China(approval No.IMB-201909-D6)on September 6,2019.
文摘c-Jun,the most extensively studied protein of the activator protein-1(AP-1)complex,is involved in numerous cell activities,such as proliferation,apoptosis,survival,tumorigenesis and tissue morphogenesis.Earlier studies focused on the structure and function have led to the identification of c-Jun as a basic leucine zipper(bZIP)transcription factor that acts as homo-or heterodimer,binding to DNA and regulating gene transcription.Later on,it was shown that extracellular signals can induce post-translational modifications of c-Jun,resulting in altered transcriptional activity and target gene expression.More recent work has uncovered multiple layers of a complex regulatory scheme in which c-Jun is able to crosstalk,amplify and integrate different signals for tissue development and disease.One example of such scheme is the autocrine amplification loop,in which signal-induced AP-1 activates the c-Jun gene promoter,while increased c-Jun expression feedbacks to potentiate AP-1 activity.Another example of such scheme,based on recent characterization of gene knockout mice,is that c-Jun integrates signals of several developmental pathways,including EGFR-ERK,EGFR-RhoA-ROCK,and activin B-MAP3K1-JNK for embryonic eyelid closure.After more than two decades of extensive research,c-Jun remains at the center stage of a molecular network with mysterious functional properties,some of which are yet to be discovered.In this article,we will provide a brief historical overview of studies on c-Jun regulation and function,and use eyelid development as an example to illustrate the complexity of c-Jun crosstalking with signaling pathways.
