[Objectives]This study was conducted to investigate characteristics of the human TCF7 L2 gene promoter.[Methods]The 2000 bp sequence of the 5’regulatory region of the human TCF7 L2 gene was obtained from the UCSC gen...[Objectives]This study was conducted to investigate characteristics of the human TCF7 L2 gene promoter.[Methods]The 2000 bp sequence of the 5’regulatory region of the human TCF7 L2 gene was obtained from the UCSC genome database.The promoter,transcription factor binding sites,CpG islands,SNPs and so on were analyzed by a variety of online softwares.[Results]The bioinformatics analysis results showed there were at least 5 potential promoters in the positive-sense strand of the 2000 bp sequence,among which-242--192 bp,-853--803 bp might contain core promoters.A TATA box and a CpG island with a length of 499 bp were found.241,944 and 1035(positive-sense strand)transcription factor binding sites were predicted by the AliBaba2.1,PROMO and JASPAR softwares,respectively.207 common transcription factor binding sites in the conserved region of human and mouse homologous TCF7 L2 gene promoter were identified with CONREAL program,involving 66 kinds of transcription factors.Two SNPs were found in the promoter region.[Conclusions]The promoter of the human TCF7 L2 gene was analyzed by bioinformatics,and the promoter characteristics were obtained.展开更多
The energy of interaction between complementary nucleotides in promoter sequences of E. coli was calculated and visualized. The graphic method for presentation of energy properties of promoter sequences was elaborated...The energy of interaction between complementary nucleotides in promoter sequences of E. coli was calculated and visualized. The graphic method for presentation of energy properties of promoter sequences was elaborated on. Data obtained indicated that energy distribution through the length of promoter sequence results in picture with minima at –35, –8 and +7 regions corresponding to areas with elevated AT (adenine-thymine) content. The most important difference from the random sequences area is related to –8. Four promoter groups and their energy properties were revealed. The promoters with minimal and maximal energy of interaction between complementary nucleotides have low strengths, the strongest promoters correspond to promoter clusters characterized by intermediate energy values.展开更多
Objective:To explore health-improving behaviors during COVID-19 pandemic and to reveal the impact of the risk perception of COVID-19 on these behaviors.Methods:We recruited a total of 510 participants for this cross-s...Objective:To explore health-improving behaviors during COVID-19 pandemic and to reveal the impact of the risk perception of COVID-19 on these behaviors.Methods:We recruited a total of 510 participants for this cross-sectional study.The data were collected online using a questionnaire booklet covering a form inquiring about the participants’demographic,physical,and mental characteristics,the Generalized Anxiety Disorder-7 Scale,and the COVID-19 Perceived Risk Scale.Results:The mean age of the participants was(28.7±10.1)years,and 76.1%were females.Of the participants,31.8%were overweight and obese.While 35.1%experienced negative alterations in their dietary patterns,23.9%reported positive changes to their diet during the pandemic.Besides,47.8%reported their sleep to be negatively affected during the pandemic.While the rate of those with extended screen time was 72.5%,it was 44.3%for the participants engaging in regular exercise.More than one-third of the participants(35.9%)had high and severe anxiety.We also found increased risk perception of COVID-19 among females,obese,those with disturbed eating and sleep quality,healthcare workers,and those with severe anxiety.Conclusions:Overall,the pandemic has appeared to have brought both positive and negative impacts on maintaining and improving eating,sleep,physical activity,and mental health.展开更多
Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such inf...Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited. In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed. DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured. Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells. Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp. The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis. The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated. Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling. Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area. In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression. H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B. In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression. Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.展开更多
三元基序家族蛋白7(tripartite motif-containing protein 7,TRIM7)作为E3泛素连接酶TRIM家族的成员,在免疫调控、代谢等生理过程中发挥重要调控作用。此外,TRIM7的异常表达与肝细胞癌(hepatocellular carcinoma,HCC)的发生发展密切相...三元基序家族蛋白7(tripartite motif-containing protein 7,TRIM7)作为E3泛素连接酶TRIM家族的成员,在免疫调控、代谢等生理过程中发挥重要调控作用。此外,TRIM7的异常表达与肝细胞癌(hepatocellular carcinoma,HCC)的发生发展密切相关并呈现出复杂的调控作用,但其在HCC中的表达调控机制尚不清楚。本研究首先利用多种在线数据库分析发现,TRIM7在HCC中高表达,并且TRIM7高表达的肝细胞癌患者预后较差;利用UCSC、JASPAR数据库分析预测TRIM7基因启动子区的转录因子结合位点。结果显示,TRIM7启动子上含有4个特异性蛋白1(specificity protein 1,SP1)转录因子结合位点。本研究利用双荧光素酶报告实验、ChIP-PCR方法检测发现,SP1通过直接结合在TRIM7启动子上的SP1结合位点,从而正调控TRIM7启动子驱动的转录活性。RT-qPCR、Western印迹检测结果进一步显示,过表达SP1在mRNA和蛋白质水平均上调TRIM7基因的表达(P<0.01),且利用SP1抑制剂Mithramycin A处理能够逆转SP1对TRIM7基因表达的调控作用(P<0.01)。总之,本研究初步揭示了TRIM7在肝细胞癌中高表达的调控机制,为深入研究该基因功能以及早期诊断、靶向治疗提供重要的理论依据。展开更多
Halobacterium halobium is a typical kind of extremely halophilic bacterium. Combined with the antibiotic resistance assay, the microcalorimetric method was used to study the promoter function of the cloned DNA fragmen...Halobacterium halobium is a typical kind of extremely halophilic bacterium. Combined with the antibiotic resistance assay, the microcalorimetric method was used to study the promoter function of the cloned DNA fragments from Halobacterium halobium J7 plasmid pHH205 in Escherichia coli TG1. The promoter probe vector, plasmid pKK232-8, was used to form the recombinants. The DNA fragment, which is the promoter for the chloramphenicol acetyl transferase （CAT） gene in plasmid pKK232-8, is about 800 bp, and the chloramphenicol resistance level presented by IC50 is about 200 μg·mL^-1, which suggests a high promoter activity. The conclusions show that there probably exist double-function or trinary-function gene promoters in Halobacterium halobium, and Archaea may contain rich genetic resources.展开更多
Targeted gene therapy has become a promising approach for lung cancer treatment.In our previous work,we reported that the targeted expression of microRNA-7(miR-7)operated by thyroid transcription factor-1(TTF-1)promot...Targeted gene therapy has become a promising approach for lung cancer treatment.In our previous work,we reported that the targeted expression of microRNA-7(miR-7)operated by thyroid transcription factor-1(TTF-1)promoter inhibited the growth of human lung cancer cells in vitro and in vivo;however,the intervention efficiency needed to be further improved.In this study,we identified the core promoter of TTF-1(from-1299 bp to-871 bp)by 5’deletion assay and screened out the putative transcription factors nuclear factor-1(NF-1)and activator protein-1(AP-1).Further analysis revealed that the expression level of NF-1,but not AP-1,was positively connected with the activation of TTF-1 core promoter in human non-small-cell lung cancer(NSCLC)cells.Moreover,the silencing of NF-1 could reduce the expression level of miR-7 operated by TTF-1 core promoter.Of note,we optimized four distinct sequences to form additional NF-1-binding sites(TGGCA)in the sequence of TTF-1 core promoter(termed asTTF-1 promoter),and verified the binding efficiency of NF-1 on theTTF-1 promoter by electrophoretic mobility shift assay(EMSA).As expected,theTTF-1 promoter could more effectively drive miR-7 expression and inhibit the growth of human NSCLC cells in vitro,accompanied by a reduced transduction of NADH dehydrogenase(ubiquinone)1αsubcomplex 4(NDUFA4)/protein kinase B(Akt)pathway.Consistently,TTF-1 promoter-driven miR-7expression could also effectively abrogate the growth and metastasis of tumor cells in a murine xenograft model of human NSCLC.Finally,no significant changes were detected in the biological indicators or the histology of some important tissues and organs,including heart,liver,and spleen.On the whole,our study revealed that the optimized TTF-1 promoter could more effectively operate miR-7 to influence the growth of human NSCLC cells,providing a new basis for the development of microRNA-based targeting gene therapy against clinical lung cancer.展开更多
基金the Diabetes Special Fund Project of Hubei University of Science and Technology(2016-18XZ12)。
文摘[Objectives]This study was conducted to investigate characteristics of the human TCF7 L2 gene promoter.[Methods]The 2000 bp sequence of the 5’regulatory region of the human TCF7 L2 gene was obtained from the UCSC genome database.The promoter,transcription factor binding sites,CpG islands,SNPs and so on were analyzed by a variety of online softwares.[Results]The bioinformatics analysis results showed there were at least 5 potential promoters in the positive-sense strand of the 2000 bp sequence,among which-242--192 bp,-853--803 bp might contain core promoters.A TATA box and a CpG island with a length of 499 bp were found.241,944 and 1035(positive-sense strand)transcription factor binding sites were predicted by the AliBaba2.1,PROMO and JASPAR softwares,respectively.207 common transcription factor binding sites in the conserved region of human and mouse homologous TCF7 L2 gene promoter were identified with CONREAL program,involving 66 kinds of transcription factors.Two SNPs were found in the promoter region.[Conclusions]The promoter of the human TCF7 L2 gene was analyzed by bioinformatics,and the promoter characteristics were obtained.
文摘The energy of interaction between complementary nucleotides in promoter sequences of E. coli was calculated and visualized. The graphic method for presentation of energy properties of promoter sequences was elaborated on. Data obtained indicated that energy distribution through the length of promoter sequence results in picture with minima at –35, –8 and +7 regions corresponding to areas with elevated AT (adenine-thymine) content. The most important difference from the random sequences area is related to –8. Four promoter groups and their energy properties were revealed. The promoters with minimal and maximal energy of interaction between complementary nucleotides have low strengths, the strongest promoters correspond to promoter clusters characterized by intermediate energy values.
文摘Objective:To explore health-improving behaviors during COVID-19 pandemic and to reveal the impact of the risk perception of COVID-19 on these behaviors.Methods:We recruited a total of 510 participants for this cross-sectional study.The data were collected online using a questionnaire booklet covering a form inquiring about the participants’demographic,physical,and mental characteristics,the Generalized Anxiety Disorder-7 Scale,and the COVID-19 Perceived Risk Scale.Results:The mean age of the participants was(28.7±10.1)years,and 76.1%were females.Of the participants,31.8%were overweight and obese.While 35.1%experienced negative alterations in their dietary patterns,23.9%reported positive changes to their diet during the pandemic.Besides,47.8%reported their sleep to be negatively affected during the pandemic.While the rate of those with extended screen time was 72.5%,it was 44.3%for the participants engaging in regular exercise.More than one-third of the participants(35.9%)had high and severe anxiety.We also found increased risk perception of COVID-19 among females,obese,those with disturbed eating and sleep quality,healthcare workers,and those with severe anxiety.Conclusions:Overall,the pandemic has appeared to have brought both positive and negative impacts on maintaining and improving eating,sleep,physical activity,and mental health.
基金We thank the Experimental Poultry Farm of the Poultry Institute,Chinese Academy of Agricultural Sciences,for providing experimental materialsThis work was supported by the Key Research and Development Program of China(2017YFE0108000)+1 种基金the National Natural Science Foundation of China(31872341,31572390)the High-Level Talent Support Program of Yangzhou University,China.
文摘Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited. In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed. DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured. Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells. Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp. The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis. The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated. Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling. Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area. In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression. H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B. In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression. Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.
文摘三元基序家族蛋白7(tripartite motif-containing protein 7,TRIM7)作为E3泛素连接酶TRIM家族的成员,在免疫调控、代谢等生理过程中发挥重要调控作用。此外,TRIM7的异常表达与肝细胞癌(hepatocellular carcinoma,HCC)的发生发展密切相关并呈现出复杂的调控作用,但其在HCC中的表达调控机制尚不清楚。本研究首先利用多种在线数据库分析发现,TRIM7在HCC中高表达,并且TRIM7高表达的肝细胞癌患者预后较差;利用UCSC、JASPAR数据库分析预测TRIM7基因启动子区的转录因子结合位点。结果显示,TRIM7启动子上含有4个特异性蛋白1(specificity protein 1,SP1)转录因子结合位点。本研究利用双荧光素酶报告实验、ChIP-PCR方法检测发现,SP1通过直接结合在TRIM7启动子上的SP1结合位点,从而正调控TRIM7启动子驱动的转录活性。RT-qPCR、Western印迹检测结果进一步显示,过表达SP1在mRNA和蛋白质水平均上调TRIM7基因的表达(P<0.01),且利用SP1抑制剂Mithramycin A处理能够逆转SP1对TRIM7基因表达的调控作用(P<0.01)。总之,本研究初步揭示了TRIM7在肝细胞癌中高表达的调控机制,为深入研究该基因功能以及早期诊断、靶向治疗提供重要的理论依据。
基金Project supported by the National Natural Science Foundation of China （Nos. 30570015, 20373051）, ＂973＂ Project （No. 2004CB719603）, Science Fund for Creative Research Group （No. 20621502 NSFC）, Science Research Foundation of Ministry of Education of China （No. [2006]8IRT0543）, and the Natural Science Foundation of Hubei Province （No. 2005ABC02）.
文摘Halobacterium halobium is a typical kind of extremely halophilic bacterium. Combined with the antibiotic resistance assay, the microcalorimetric method was used to study the promoter function of the cloned DNA fragments from Halobacterium halobium J7 plasmid pHH205 in Escherichia coli TG1. The promoter probe vector, plasmid pKK232-8, was used to form the recombinants. The DNA fragment, which is the promoter for the chloramphenicol acetyl transferase （CAT） gene in plasmid pKK232-8, is about 800 bp, and the chloramphenicol resistance level presented by IC50 is about 200 μg·mL^-1, which suggests a high promoter activity. The conclusions show that there probably exist double-function or trinary-function gene promoters in Halobacterium halobium, and Archaea may contain rich genetic resources.
基金the National Natural Science Foundation of China(Nos.32160178,82160503,31760258,and 81960509)the Project of the Guizhou Provincial Department of Science and Technology(Nos.QKH-JC-2018-1428,QKHZC-2020-4Y156,and QKH-JC-ZK-2022-624)+1 种基金the Collaborative Innovation Center of Chinese Ministry of Education(No.2020-39)the Program for Excellent Young Talents of Zunyi Medical University(No.15ZY-001),China。
文摘Targeted gene therapy has become a promising approach for lung cancer treatment.In our previous work,we reported that the targeted expression of microRNA-7(miR-7)operated by thyroid transcription factor-1(TTF-1)promoter inhibited the growth of human lung cancer cells in vitro and in vivo;however,the intervention efficiency needed to be further improved.In this study,we identified the core promoter of TTF-1(from-1299 bp to-871 bp)by 5’deletion assay and screened out the putative transcription factors nuclear factor-1(NF-1)and activator protein-1(AP-1).Further analysis revealed that the expression level of NF-1,but not AP-1,was positively connected with the activation of TTF-1 core promoter in human non-small-cell lung cancer(NSCLC)cells.Moreover,the silencing of NF-1 could reduce the expression level of miR-7 operated by TTF-1 core promoter.Of note,we optimized four distinct sequences to form additional NF-1-binding sites(TGGCA)in the sequence of TTF-1 core promoter(termed asTTF-1 promoter),and verified the binding efficiency of NF-1 on theTTF-1 promoter by electrophoretic mobility shift assay(EMSA).As expected,theTTF-1 promoter could more effectively drive miR-7 expression and inhibit the growth of human NSCLC cells in vitro,accompanied by a reduced transduction of NADH dehydrogenase(ubiquinone)1αsubcomplex 4(NDUFA4)/protein kinase B(Akt)pathway.Consistently,TTF-1 promoter-driven miR-7expression could also effectively abrogate the growth and metastasis of tumor cells in a murine xenograft model of human NSCLC.Finally,no significant changes were detected in the biological indicators or the histology of some important tissues and organs,including heart,liver,and spleen.On the whole,our study revealed that the optimized TTF-1 promoter could more effectively operate miR-7 to influence the growth of human NSCLC cells,providing a new basis for the development of microRNA-based targeting gene therapy against clinical lung cancer.
