Objective To investigate the cell-type-specific enhancer (CTSE) in HPV16 and its variation in cervical carcinoma. Methods CTSEs were detected by polymerase chain reaction (PCR) in 58 cervical carcinoma from Shaanxi pr...Objective To investigate the cell-type-specific enhancer (CTSE) in HPV16 and its variation in cervical carcinoma. Methods CTSEs were detected by polymerase chain reaction (PCR) in 58 cervical carcinoma from Shaanxi province; in addition variation of CTSEs was analyzed through single-strand conformation polymorphisms (SSCP). Results HPV16 CTSEs were detectable in 34 of 58 (57%) specimens and mutant rate was 41%(14/34) and the main mutations of chosen randomly variant CTSE (CTSEv) happened at YY1 binding sites in addition to glucocoticoid response elements (GRE). Conclusion CTSE in some specimens of Shaanxi province was obviously different from that in HPV16 wild type and variant CTSE might affect the transcriptional regulation of LCR on viral P97, which regulates over-expression of viral oncogenes in cervical carcinoma.展开更多
The CD80 （B7-1） costimulatory molecule is expressed on the surface of B cells and its expression is transcriptionally upregulated upon stimuli. To identify the region of murine CD80 promoter that direct cell type sp...The CD80 （B7-1） costimulatory molecule is expressed on the surface of B cells and its expression is transcriptionally upregulated upon stimuli. To identify the region of murine CD80 promoter that direct cell type specific gene expression, four promoters construct of CD80 gene were generated with DNA fragments fused to the GFP reporter gene. In the present study, significant promoter activity was detected with all four promoter constructs only in the murine B lymphocyte. Further, the CD80 promoter region extending from -3,005 bp to ＋273 bp in relation to the previously reported transcription start site, was identified as tissue specific region. Interestingly, the shortest 700 bp （-427/＋273） of promoter fragment was sufficient to direct the CD80 gene expression. The level of CD80 expression was also found to be modulated by exogenous stimuli in B lymphocyte. Additionally, it was demonstrated that the CD80 gene expression is regulated at the level of transcription where the inducible CD80 gene transcript was detected in B lymphocyte with increasing time.展开更多
Behavioral and molecular characterization of cell-type specific populations governing fear learning and behavior is a promising avenue for the rational identification of potential therapeutics for fear-related disorde...Behavioral and molecular characterization of cell-type specific populations governing fear learning and behavior is a promising avenue for the rational identification of potential therapeutics for fear-related disorders.Identification of cell-type specific changes in neuronal translation following fear learning allows for targeted pharmacological intervention during fear extinction learning,mirroring possible treatment strategies in humans.Here we identify the central amygdala(Ce A)Drd2-expressing population as a fear-supporting population that is molecularly distinct from other,previously identified fear-supporting CeA populations.Sequencing of actively translating transcripts of Drd2 neurons identifies m RNAs that are differentially regulated following fear learning including Npy5r,Rxrg,Sst5r,Fgf3,Erb B4,Fkbp14,Dlk1,Ssh3 and Adora2a.Direct pharmacological manipulation of NPY5R,RXR,and ADORA2A confirms their importance in fear behavior and validates the present approach of identifying pharmacological targets for the modulation of emotional learning.展开更多
INTRODUCTIONHepatitis B virus (HBV) is the most commonetiologic agent for infectious liver diseases. It isestimated that there are more than 250 millionchronic HBV carriersin the world today and thereis a significant ...INTRODUCTIONHepatitis B virus (HBV) is the most commonetiologic agent for infectious liver diseases. It isestimated that there are more than 250 millionchronic HBV carriersin the world today and thereis a significant association among persistentinfection, liver cirrhosis and hepatocellularcarcinoma[1-3].展开更多
To study transient tissue specific linalool emission and to examine its fate, plastid targeted dual linalool/nerolidol synthase (FaNES1) was introduced into Nicotiana benthamiana leaves under LTP1, SUC2, RbcS and CaMV...To study transient tissue specific linalool emission and to examine its fate, plastid targeted dual linalool/nerolidol synthase (FaNES1) was introduced into Nicotiana benthamiana leaves under LTP1, SUC2, RbcS and CaMV35 promoters. Tissue specificity of the promoters was tested with β-glucuronidase (GUS) reporter gene. Promoter::GUS/FaNES1 fusion was confirmed with colony PCR and agro-infiltrated into six weeks old N. benthamiana leaves. Eight days post inoculation (dpi), promoter::GUS constructs were examined with GUS histochemical staining at 1.5 h, 3.5 h, 5.5 h and 16 h incubation times. After 4/8 days, FaNES1 construct agro-inoculated leaves were assessed for linalool emissions and its conjugates using a dynamic headspace system and LC-MS, respectively. There was high affinity of promoters to their respective cell-types although it was not as specific as in stable transformation. This is possibly due to activations of many copies of transiently introduced promotes by few transcription factors of the respective promoter that naturally exist in untargeted cell-types. GUS staining intensity was stronger in leaf veins and injured sites compared to other plant tissues under all heterologous promoters, and it was gradually increased with increase in incubation times that could be explained by promoters wounding responses and/ or GUS leakage to unstained sites. Linalool emission was obtained in the same pattern under all promoters;it was higher at day 4 than day 8. Its concentration declined 44, 12, 4.5 and 4 folds at 8 dpi under LTP1, SUC2, RbcS and CaMV35S promoters, respectively. Conversely, linalool conjugates were significantly increased at day 8. These might be due to T-DNA degradations and/or protein modifications 4 dpi. LTP1 promoter was the least efficient to drive both GUS and FaNES1 possibly due to immature plastids in epidermal cells and/or its weak performance. Hence, to study FaNES1 activity in transient assay it is suggested to use relatively shorter duration and longer inoculation times for linalool and its conjugates, respectively.展开更多
Studies on cell signaling pay more attention to spatial dynamics and how such diverse organization can relate to high order of cellular capabilities.To overview the specificity of cell signaling,we integrated human re...Studies on cell signaling pay more attention to spatial dynamics and how such diverse organization can relate to high order of cellular capabilities.To overview the specificity of cell signaling,we integrated human receptome data with proteome spatial expression profiles to systematically investigate the specificity of receptors and receptor-triggered transduction networks across 62 normal cell types and 14 cancer types.Six percent receptors showed cell-type-specific expression,and 4% signaling networks presented enriched cell-specific proteins induced by the receptors.We introduced a concept of“response context”to annotate the cell-type dependent signaling networks.We found that most cells respond similarly to the same stimulus,as the“response contexts”presented high functional similarity.Despite this,the subtle spatial diversity can be observed from the difference in network architectures.The architecture of the signaling networks in nerve cells displayed less completeness than that in glandular cells,which indicated cellular-context dependent signaling patterns are elaborately spatially organized.Likewise,in cancer cells most signaling networks were generally dysfunctional and less complete than that in normal cells.However,glioma emerged hyper-activated transduction mechanism in malignant state.Receptor ATP6AP2 and TNFRSF21 induced rennin-angiotensin and apoptosis signaling were found likely to explain the glioma-specific mechanism.This work represents an effort to decipher context-specific signaling network from spatial dimension.Our results indicated that although a majority of cells engage general signaling response with subtle differences,the spatial dynamics of cell signaling can not only deepen our insights into different signaling mechanisms,but also help understand cell signaling in disease.展开更多
目的:确定前列腺癌中Sp1通过PKM2途径对前列腺癌细胞的增殖和代谢的作用。方法:前列腺癌细胞株DU145与PC3体外转染Sp1 si RNA与阴性对照无意义核苷酸序列(NC组),采用葡萄糖检测试剂盒与乳酸检测试剂盒检测转染后有氧糖酵解变化;CCK-8实...目的:确定前列腺癌中Sp1通过PKM2途径对前列腺癌细胞的增殖和代谢的作用。方法:前列腺癌细胞株DU145与PC3体外转染Sp1 si RNA与阴性对照无意义核苷酸序列(NC组),采用葡萄糖检测试剂盒与乳酸检测试剂盒检测转染后有氧糖酵解变化;CCK-8实验检测转染后的细胞增殖;q RT-PCR检测转染后PKM2表达;Western Blotting检测转染后Sp1与PKM2表达。结果:DU145与PC3转染Sp1 si RNA后与NC组比较,剩余葡萄糖显著偏高,乳酸产生显著下降;CCK8实验表明抑制Sp1后能显著抑制前列腺癌细胞的增殖能力;q RT-PCR实验显示抑制Sp1后明显抑制PKM2的表达;Western Blotting显示转染的Sp1 si RNA对Sp1具有较高的沉默效率,且PKM2的表达也降低。结论:Sp1在前列腺癌细胞株DU145与PC3中可通过直接作用于PKM2促进细胞代谢。展开更多
目的:检测食管鳞癌组织中人乳头瘤病毒(HPV)16E6、E7基因和细胞特异性增强子片段(CTSE)。方法:采用聚合酶链反应法(PCR)检测40例食管鳞癌和20例正常食管黏膜中HPV16E6、E7基因和病毒长控制区内(long control region,LCR)的细胞型特异性...目的:检测食管鳞癌组织中人乳头瘤病毒(HPV)16E6、E7基因和细胞特异性增强子片段(CTSE)。方法:采用聚合酶链反应法(PCR)检测40例食管鳞癌和20例正常食管黏膜中HPV16E6、E7基因和病毒长控制区内(long control region,LCR)的细胞型特异性增强子(cell type specific enhancer,CTSE)。结果:在40例食管鳞癌中,HPV16的E6、E7基因和CTSE片段的检出率分别是37.5%(15/40)、42.5%(17/40)和40%(16/40),20例正常食管黏膜中E6、E7和CTSE的检出率分别为0%(0/20)、0%(0/20)和5%(1/20),两者均存在显著性差异(P<0.05)。CTSE片段分别与E6和E7基因有明显相关性(P<0.05)。食管鳞癌中E6、E7基因及CTSE的检出率在患者不同性别、年龄、肿瘤浸润程度、淋巴结转移和组织学分级肿瘤中差异均无显著性(P>0.05)。结论:E6和E7基因与CTSE片段共存于HPV16感染的食管鳞癌组织中,三者可能与食管鳞癌的发生和发展有关。展开更多
目的探讨联合监测血浆尿激酶型纤溶酶原激活因子(urokinase type plasminogen activator,uPA)系统与血清肿瘤标记物神经原特异性烯醇化酶(neuron specific enolase,NSE)在小细胞肺癌(small cell lungcancer,SCLC)诊断、治疗和预后判断...目的探讨联合监测血浆尿激酶型纤溶酶原激活因子(urokinase type plasminogen activator,uPA)系统与血清肿瘤标记物神经原特异性烯醇化酶(neuron specific enolase,NSE)在小细胞肺癌(small cell lungcancer,SCLC)诊断、治疗和预后判断的临床价值。方法采用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测48例SCLC患者和37例健康者血浆uPA及其受体(urokinase type plasminogen activator receptor,uPAR)、抑制因子-1(plasminogen activator inhibitor-1,PAI-1)的水平及血清肿瘤标记物NSE含量,并进行统计学分析。结果 SCLC患者血浆uPA、uPAR、PAI-1水平和血清NSE含量均高于正常对照组,差异有统计学意义(P<0.05)。有淋巴结转移和远处转移患者uPA、uPAR、PAI-1和NSE水平明显高于无转移患者;Ⅲ、Ⅳ期患者uPA、uPAR、PAI-1和NSE水平高于Ⅰ、Ⅱ期患者;治疗前uPA、uPAR、PAI-1及NSE水平高于治疗后;uPAR和NSE水平在各生存组间有明显差异,差异均有统计学意义(P<0.05)。血浆uPA、uPAR、PAI-1水平之间及uPAR与NSE水平之间均呈直线相关(P<0.05)。结论同血清NSE一样,血浆uPA、uPAR和PAI-1水平可作为肿瘤标记物,用于SCLC的诊断、治疗及预后判断;血浆uPAR水平和血清NSE含量的联合检测对SCLC的预后诊断有重要临床意义。展开更多
基金The study was supported by National Natural Science Foundation of China (No.39970035) .
文摘Objective To investigate the cell-type-specific enhancer (CTSE) in HPV16 and its variation in cervical carcinoma. Methods CTSEs were detected by polymerase chain reaction (PCR) in 58 cervical carcinoma from Shaanxi province; in addition variation of CTSEs was analyzed through single-strand conformation polymorphisms (SSCP). Results HPV16 CTSEs were detectable in 34 of 58 (57%) specimens and mutant rate was 41%(14/34) and the main mutations of chosen randomly variant CTSE (CTSEv) happened at YY1 binding sites in addition to glucocoticoid response elements (GRE). Conclusion CTSE in some specimens of Shaanxi province was obviously different from that in HPV16 wild type and variant CTSE might affect the transcriptional regulation of LCR on viral P97, which regulates over-expression of viral oncogenes in cervical carcinoma.
文摘The CD80 （B7-1） costimulatory molecule is expressed on the surface of B cells and its expression is transcriptionally upregulated upon stimuli. To identify the region of murine CD80 promoter that direct cell type specific gene expression, four promoters construct of CD80 gene were generated with DNA fragments fused to the GFP reporter gene. In the present study, significant promoter activity was detected with all four promoter constructs only in the murine B lymphocyte. Further, the CD80 promoter region extending from -3,005 bp to ＋273 bp in relation to the previously reported transcription start site, was identified as tissue specific region. Interestingly, the shortest 700 bp （-427/＋273） of promoter fragment was sufficient to direct the CD80 gene expression. The level of CD80 expression was also found to be modulated by exogenous stimuli in B lymphocyte. Additionally, it was demonstrated that the CD80 gene expression is regulated at the level of transcription where the inducible CD80 gene transcript was detected in B lymphocyte with increasing time.
文摘Behavioral and molecular characterization of cell-type specific populations governing fear learning and behavior is a promising avenue for the rational identification of potential therapeutics for fear-related disorders.Identification of cell-type specific changes in neuronal translation following fear learning allows for targeted pharmacological intervention during fear extinction learning,mirroring possible treatment strategies in humans.Here we identify the central amygdala(Ce A)Drd2-expressing population as a fear-supporting population that is molecularly distinct from other,previously identified fear-supporting CeA populations.Sequencing of actively translating transcripts of Drd2 neurons identifies m RNAs that are differentially regulated following fear learning including Npy5r,Rxrg,Sst5r,Fgf3,Erb B4,Fkbp14,Dlk1,Ssh3 and Adora2a.Direct pharmacological manipulation of NPY5R,RXR,and ADORA2A confirms their importance in fear behavior and validates the present approach of identifying pharmacological targets for the modulation of emotional learning.
基金Project supported by the grant from Science Foundation of Ministry of Health of China, No. 96-1-347.
文摘INTRODUCTIONHepatitis B virus (HBV) is the most commonetiologic agent for infectious liver diseases. It isestimated that there are more than 250 millionchronic HBV carriersin the world today and thereis a significant association among persistentinfection, liver cirrhosis and hepatocellularcarcinoma[1-3].
文摘To study transient tissue specific linalool emission and to examine its fate, plastid targeted dual linalool/nerolidol synthase (FaNES1) was introduced into Nicotiana benthamiana leaves under LTP1, SUC2, RbcS and CaMV35 promoters. Tissue specificity of the promoters was tested with β-glucuronidase (GUS) reporter gene. Promoter::GUS/FaNES1 fusion was confirmed with colony PCR and agro-infiltrated into six weeks old N. benthamiana leaves. Eight days post inoculation (dpi), promoter::GUS constructs were examined with GUS histochemical staining at 1.5 h, 3.5 h, 5.5 h and 16 h incubation times. After 4/8 days, FaNES1 construct agro-inoculated leaves were assessed for linalool emissions and its conjugates using a dynamic headspace system and LC-MS, respectively. There was high affinity of promoters to their respective cell-types although it was not as specific as in stable transformation. This is possibly due to activations of many copies of transiently introduced promotes by few transcription factors of the respective promoter that naturally exist in untargeted cell-types. GUS staining intensity was stronger in leaf veins and injured sites compared to other plant tissues under all heterologous promoters, and it was gradually increased with increase in incubation times that could be explained by promoters wounding responses and/ or GUS leakage to unstained sites. Linalool emission was obtained in the same pattern under all promoters;it was higher at day 4 than day 8. Its concentration declined 44, 12, 4.5 and 4 folds at 8 dpi under LTP1, SUC2, RbcS and CaMV35S promoters, respectively. Conversely, linalool conjugates were significantly increased at day 8. These might be due to T-DNA degradations and/or protein modifications 4 dpi. LTP1 promoter was the least efficient to drive both GUS and FaNES1 possibly due to immature plastids in epidermal cells and/or its weak performance. Hence, to study FaNES1 activity in transient assay it is suggested to use relatively shorter duration and longer inoculation times for linalool and its conjugates, respectively.
基金kindly funded by National Natural Science Foundation of China(Grant No.31070752)in part supported by the National Basic Research Program(973 Program)(Nos 2011CB910204,2010CB529206 and 2010CB912702)+4 种基金Key Infectious Disease Project(No.2012ZX10002012-014)Research Program of Chinese Academy of Sciences(Nos.KSCX2-EW-R-04,KSCX2-YW-R-190 and 2011KIP204)National Natural Science Foundation of China(Grant No.30900272)Chinese Ministry for Science and Technology Grant(No.2008BAI64B01)the National High Technology Research and Development Program(863 Program)(No.2009AA02Z304).
文摘Studies on cell signaling pay more attention to spatial dynamics and how such diverse organization can relate to high order of cellular capabilities.To overview the specificity of cell signaling,we integrated human receptome data with proteome spatial expression profiles to systematically investigate the specificity of receptors and receptor-triggered transduction networks across 62 normal cell types and 14 cancer types.Six percent receptors showed cell-type-specific expression,and 4% signaling networks presented enriched cell-specific proteins induced by the receptors.We introduced a concept of“response context”to annotate the cell-type dependent signaling networks.We found that most cells respond similarly to the same stimulus,as the“response contexts”presented high functional similarity.Despite this,the subtle spatial diversity can be observed from the difference in network architectures.The architecture of the signaling networks in nerve cells displayed less completeness than that in glandular cells,which indicated cellular-context dependent signaling patterns are elaborately spatially organized.Likewise,in cancer cells most signaling networks were generally dysfunctional and less complete than that in normal cells.However,glioma emerged hyper-activated transduction mechanism in malignant state.Receptor ATP6AP2 and TNFRSF21 induced rennin-angiotensin and apoptosis signaling were found likely to explain the glioma-specific mechanism.This work represents an effort to decipher context-specific signaling network from spatial dimension.Our results indicated that although a majority of cells engage general signaling response with subtle differences,the spatial dynamics of cell signaling can not only deepen our insights into different signaling mechanisms,but also help understand cell signaling in disease.
文摘目的:确定前列腺癌中Sp1通过PKM2途径对前列腺癌细胞的增殖和代谢的作用。方法:前列腺癌细胞株DU145与PC3体外转染Sp1 si RNA与阴性对照无意义核苷酸序列(NC组),采用葡萄糖检测试剂盒与乳酸检测试剂盒检测转染后有氧糖酵解变化;CCK-8实验检测转染后的细胞增殖;q RT-PCR检测转染后PKM2表达;Western Blotting检测转染后Sp1与PKM2表达。结果:DU145与PC3转染Sp1 si RNA后与NC组比较,剩余葡萄糖显著偏高,乳酸产生显著下降;CCK8实验表明抑制Sp1后能显著抑制前列腺癌细胞的增殖能力;q RT-PCR实验显示抑制Sp1后明显抑制PKM2的表达;Western Blotting显示转染的Sp1 si RNA对Sp1具有较高的沉默效率,且PKM2的表达也降低。结论:Sp1在前列腺癌细胞株DU145与PC3中可通过直接作用于PKM2促进细胞代谢。
文摘目的:检测食管鳞癌组织中人乳头瘤病毒(HPV)16E6、E7基因和细胞特异性增强子片段(CTSE)。方法:采用聚合酶链反应法(PCR)检测40例食管鳞癌和20例正常食管黏膜中HPV16E6、E7基因和病毒长控制区内(long control region,LCR)的细胞型特异性增强子(cell type specific enhancer,CTSE)。结果:在40例食管鳞癌中,HPV16的E6、E7基因和CTSE片段的检出率分别是37.5%(15/40)、42.5%(17/40)和40%(16/40),20例正常食管黏膜中E6、E7和CTSE的检出率分别为0%(0/20)、0%(0/20)和5%(1/20),两者均存在显著性差异(P<0.05)。CTSE片段分别与E6和E7基因有明显相关性(P<0.05)。食管鳞癌中E6、E7基因及CTSE的检出率在患者不同性别、年龄、肿瘤浸润程度、淋巴结转移和组织学分级肿瘤中差异均无显著性(P>0.05)。结论:E6和E7基因与CTSE片段共存于HPV16感染的食管鳞癌组织中,三者可能与食管鳞癌的发生和发展有关。
