In plants, the phloem is the component of the vascular system that delivers nutrients and transmits signals from mature leaves to developing sink tissues. Recent studies have identified proteins, mRNA, and small RNA w...In plants, the phloem is the component of the vascular system that delivers nutrients and transmits signals from mature leaves to developing sink tissues. Recent studies have identified proteins, mRNA, and small RNA within the phloem sap of several plant species. It is now of considerable interest to elucidate the biological functions of these potential long-distance signal agents, to further our understanding of how plants coordinate their developmental programs at the whole-plant level. In this study, we developed a strategy for the functional analysis of phloem-mobile mRNA by focusing on IAA transcripts, whose mobility has previously been reported in melon （Cucumis melo cv. Hale＇s Best Jumbo）. Indoleacetic acid （IAA） proteins are key transcriptional regulators of auxin signaling, and are involved in a broad range of developmental processes including root development. We used a combination of vasculature-enriched sampling and hetero-grafting techniques to identify IAA18 and IAA28 as phloemmobile transcripts in the model plant Arabidopsis thaliana. Micro-grafting experiments were used to confirm that these IAA transcripts, which are generated in vascular tissues of mature leaves, are then transported into the root system where they negatively regulate lateral root formation. Based on these findings, we present a model in which auxin distribution, in combination with phloem-mobile Aux/IAA transcripts, can determine the sites of auxin action.展开更多
Plant metabolomics developed as a powerful tool to examine gene functions and to gain deeper insight into the physiology of the plant cell. In this study, we screened Arabidopsis lines overexpressing rice full-length ...Plant metabolomics developed as a powerful tool to examine gene functions and to gain deeper insight into the physiology of the plant cell. In this study, we screened Arabidopsis lines overexpressing rice full-length （FL） cDNAs （rice FOX Arabidopsis lines） using a gas chromatography-time-of-flight mass spectrometry （GC-TOF/MS）-based technique to identify rice genes that caused metabolic changes. This screening system allows fast and reliable identification of candi- date lines showing altered metabolite profiles. We performed metabolomic and transcriptomic analysis of a rice FOX Ara- bidopsis line that harbored the FL cDNA of the rice ortholog of the Lateral Organ Boundaries （LOB） Domain （LBD）/ Asymmetric Leaves2-1ike （ASL） gene of Arabidopsis, At-LBD37/ASL39. The investigated rice FOX Arabidopsis line showed prominent changes in the levels of metabolites related to nitrogen metabolism. The transcriptomic data as well as the results from the metabolite analysis of the Arabidopsis At-LBD37/ASL39-overexpressor plants were consistent with these findings. Furthermore, the metabolomic and transcriptomic analysis of the Os-LBD37/ASL39-overexpressing rice plants indicated that Os-LBD37/ASL39 is associated with processes related to nitrogen metabolism in rice. Thus, the combination of a metabolomics-based screening method and a gain-of-function approach is useful for rapid characterization of novel genes in both Arabidopsis and rice.展开更多
Silver nanoparticles （AgNPs） are one of the most widely used nanomaterials, but the mechanism of AgNP toxicity in terrestrial plants is still unclear. We compared the toxic effects of AgNPs and Ag＋ on Arabidopsis t...Silver nanoparticles （AgNPs） are one of the most widely used nanomaterials, but the mechanism of AgNP toxicity in terrestrial plants is still unclear. We compared the toxic effects of AgNPs and Ag＋ on Arabidopsis thaliana at the physiological, ultrastructural and molecular levels. AgNPs did not affect seed germination; however, they showed stronger inhibitory effect on root elongation than Ag＋ . The results of transmission electron microscopy and metal content analysis showed that AgNPs could be accumulated in leaves. These absorbed AgNPs disrupted the thylakoid membrane structure and decreased chlorophyll content, which can inhibit plant growth. By comparison, a small amount of Ag＋ was absorbed by seedlings, and it did not pronouncedly affect chloroplast structure and other metal ion absorption as AgNPs did. Compared with Ag＋ , AgNPs could alter the transcription of antioxidant and aquaporin genes, indicating that AgNPs changed the balance between the oxidant and antioxidant systems, and also affected the homeostasis of water and other small molecules within the plant body. All the data from physiological, ultrastructural and molecular levels suggest that AgNPs were more toxic than Ag＋ .展开更多
Background:Gastric cancer(GC)is one of the most common malignancies worldwide,particularly in China.DNA damage-inducible transcript 4(DDIT4)is a mammalian target of rapamycin inhibitor and is induced by various cellul...Background:Gastric cancer(GC)is one of the most common malignancies worldwide,particularly in China.DNA damage-inducible transcript 4(DDIT4)is a mammalian target of rapamycin inhibitor and is induced by various cellular stresses;however,its critical role in GC remains poorly understood.The present study aimed to investigate the poten-tial relationship and the underlying mechanism between DDIT4 and GC development.Methods:We used western blotting,real-time polymerase chain reaction,and immunohistochemical or immunoflu-orescence to determine DDIT4 expression in GC cells and tissues.High-content screening,cell counting kit-8 assays,colony formation,and in vivo tumorigenesis assays were performed to evaluate cell proliferation.Flow cytometry was used to investigate cell apoptosis and cell cycle distribution.Results:DDIT4 was upregulated in GC cells and tissue.Furthermore,downregulating DDIT4 in GC cells inhibited proliferation both in vitro and in vivo and increased 5-fluorouracil-induced apoptosis and cell cycle arrest.In contrast,ectopic expression of DDIT4 in normal gastric epithelial cells promoted proliferation and attenuated chemosensitivity.Further analysis indicated that the mitogen-activated protein kinase and p53 signaling pathways were involved in the suppression of proliferation,and increased chemosensitivity upon DDIT4 downregulation.Conclusion:DDIT4 promotes GC proliferation and tumorigenesis,providing new insights into the role of DDIT4 in the tumorigenesis of human GC.展开更多
The plasma membrane Na＋/H＋-antiporter salt overly sensitive1 （SOS1） from the halophytic Arabidopsis-relative Thellungiella halophila （ThSOS1） shows conserved sequence and domain structure with the orthologous ge...The plasma membrane Na＋/H＋-antiporter salt overly sensitive1 （SOS1） from the halophytic Arabidopsis-relative Thellungiella halophila （ThSOS1） shows conserved sequence and domain structure with the orthologous genes from Arabidopsis thaliana and other plants. When expression of ThSOSt was reduced by RNA interference （RNAi）, pronounced characteristics of salt-sensitivity were observed. We were interested in monitoring altered transcriptional responses between Thellungiella wild type and thsost-4, a representative RNAi line with particular emphasis on root responses to salt stress at 350 mmol/L NaCI, a concentration that is only moderately stressful for mature wild type plants. Transcript profiling revealed several functional categories of genes that were differently affected in wild-type and RNAi plants. Down-regulation of SOS1 resulted in different gene expression even in the absence of stress. The pattern of gene induction in the RNAi plant under salt stress was similar to that of glycophytic Arabidopsis rather than that of wild type Thellungiella. The RNAi plants failed to down-regulate functions that are normally reduced in wild type Thellungiella upon stress and did not up-regulate functions that characterize the Thellungiella salt stress response. Metabolite changes observed in wild type Thellungiella after salt stress were less pronounced or absent in RNAi plants. Transcript and metabolite behavior suggested SOS1 functions including but also extending its established function as a sodium transporter. The down-regulation of ThSOS1 converted the halophyte Thellungiella into a salt-sensitive plant.展开更多
Background Folic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of fotic acid performing its biological function. Therefore, the dysfunction of di...Background Folic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of fotic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase （DHFR） gene knock-down zebrafish embryos, investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 （HAND2） and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations. Methods Morpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression. Results DHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos （P〈0.05）. Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos. Conclusions DHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation.展开更多
AIM To clarify the mechanisms of HOX transcript antisense intergenic RNA(HOTAIR) in gastric cancer(GC) migration and invasion.METHODS Quantitative real-time polymerase chain reaction(qp CR) was used to detect the expr...AIM To clarify the mechanisms of HOX transcript antisense intergenic RNA(HOTAIR) in gastric cancer(GC) migration and invasion.METHODS Quantitative real-time polymerase chain reaction(qp CR) was used to detect the expression level of HOTAIR in GC tissues. The correlation of its expression with clinicopathological features was analyzed. Area under receiver operating characteristic curve(AUCROC) was constructed to evaluate the diagnostic value of HOTAIR. Wound-healing assay and Transwell assay were performed to detect the biological effects of HOTAIR in GC cells. qp CR,western blot and immunohistochemistry were used to evaluate the m RNA and protein expression of E-cadherin. RNAbinding protein immunoprecipitation was used for the analysis of EZH2 interactions with HOTAIR. Chromatin immunoprecipitation assay was performed to investigate direct interactions between EZH2 and E-cadherin.RESULTS The expression of HOTAIR was up-regulated in GC tumorous tissues compared with the para-tumorous tissues(p < 0.001). Its over-expression was correlated with tumor-node-metastasis(TNM) stage(p = 0.024),tumor invasion(p = 0.018),lymph node metastasis(p = 0.023),and poor prognosis(p < 0.001). Multivariate Cox regression analysis confirmed expression of HOTAIR as an independent predictor of overall survival(p = 0.033),together with TNM stage(p = 0.002) and lymph node metastasis(p = 0.002). The AUCROC was up to 0.709(95%CI: 0.623-0.785,p < 0.001). Knockdown of HOTAIR by si RNA in GC cells suppressed the migration and invasion of GC cells. Significantly negative correlation between HOTAIR and E-cadherin was found in GC tissues and cell lines,and HOTAIR contributed to the regulation of E-cadherin through binding to EZH2 with the E-cadherin promoter. CONCLUSION HOTAIR may play a pivotal role in tumor cell migration and invasion. It can be used as a potential diagnostic and prognostic biomarker for GC.展开更多
Objective： To study the expression level and role of apoptosis-associated speck-like protein containing a caspase recruitment domain （PYCARD） gene transcript variant mRNA in peripheral blood mononuclear cells （PB...Objective： To study the expression level and role of apoptosis-associated speck-like protein containing a caspase recruitment domain （PYCARD） gene transcript variant mRNA in peripheral blood mononuclear cells （PBMCs） of primary gout （PG） patients with different Chinese medicine （CM） syndromes. Methods： The expressions of PYCARD gene transcript variant mRNA and interleukin-1β （IL-1β） mRNA in PBMCs were investigated in 96 PG patients with acute phase （APPG, 44 cases） and non-acute phase （NAPPG, 52 cases） and 30 healthy controls （HCs） by reverse transcription-polymerase chain reaction （PCR） and/or real-time quantitative PCR. PYCARD and nuclear factor-κB （p50） [NF-κB （p50）] protein was detected by Western blot in PBMCs respectively. IL-1β, IL-4 and IL-10 protein levels in plasma of HCs and PG patients were measured by enzyme-linked immuno sorbent assay. Results： The main CM syndromes in APPG patients were obstruction of dampness and heat syndrome （ODHS, 36.36%） and intermingled phlegm-blood stasis syndrome （IPBSS, 27.27%）, while in NAPPG patients were Pi （Spleen）-deficiency induced dampness syndrome （PDIDS, 40.38%） and qi-blood deficiency syndrome （QBDS, 26.92%）. It showed statistical significances of the expressions of PYCARD gene and its transcript variant mRNA, the protein of PYCARD and NF-κB （p50） and the plasma IL-1β, IL-4 and IL-10 in APPG, NAPPG, ODHS, IPBSS, PDIDS and QBDS groups, compared with the HC group respectively （P〈0.05 or P〈0.01）. There were also significant differences of mRNA expressions of PYCARD-1 and PYCARD-2 as well as protein expressions of IL-1β, IL-4 and IL-10 among the 4 CM syndromes groups （P〈0.05 or P〈0.01）. Correlation analysis showed positive correlation between the mRNA expressions of PYCARD-1 gene transcript variant and IL-1β in APPG patients （r=0.3088, P=0.0183）. Conclusion： PYCARD gene and its transcript variant may play a critical and regulative role in the inflammatory response展开更多
BACKGROUND Anti-programmed death therapy has thrust immunotherapy into the spotlight.However,such therapy has a modest response in hepatocellular carcinoma(HCC).Epigenetic immunomodulation is a suggestive combinatoria...BACKGROUND Anti-programmed death therapy has thrust immunotherapy into the spotlight.However,such therapy has a modest response in hepatocellular carcinoma(HCC).Epigenetic immunomodulation is a suggestive combinatorial therapy with immune checkpoint blockade.Non-coding ribonucleic acid(ncRNA)driven regulation is a major mechanism of epigenetic modulation.Given the wide range of ncRNAs that co-opt in programmed cell-death protein 1(PD-1)/programmed death ligand 1(PD-L1)regulation,and based on the literature,we hypothesized that miR-155-5p,miR-194-5p and long non-coding RNAs(lncRNAs)X-inactive specific transcript(XIST)and MALAT-1 are involved in a regulatory upstream pathway for PD-1/PD-L1.Recently,nutraceutical therapeutics in cancers have received increasing attention.Thus,it is interesting to study the impact of oleuropein on the respective study key players.AIM To explore potential upstream regulatory ncRNAs for the immune checkpoint PD-1/PD-L1.METHODS Bioinformatics tools including microrna.org and lnCeDB software were adopted to detect targeting of miR-155-5p,miR-194-5p and lncRNAs XIST and MALAT-1 to PD-L1 mRNA,respectively.In addition,Diana tool was used to predict targeting of both aforementioned miRNAs to lncRNAs XIST and MALAT-1.HCC and normal tissue samples were collected for scanning of PD-L1,XIST and MALAT-1 expression.To study the interaction among miR-155-5p,miR-194-5p,lncRNAs XIST and MALAT-1,as well as PD-L1 mRNA,a series of transfections of the Huh-7 cell line was carried out.RESULTS Bioinformatics software predicted that miR-155-5p and miR-194-5p can target PDL1,MALAT-1 and XIST.MALAT-1 and XIST were predicted to target PD-L1 mRNA.PD-L1 and XIST were significantly upregulated in 23 HCC biopsies compared to healthy controls;however,MALAT-1 was barely detected.MiR-194 induced expression elevated the expression of PD-L1,XIST and MALAT-1.However,overexpression of miR-155-5p induced the upregulation of PD-L1 and XIST,while it had a negative impact on MALAT-1 expression.Knockdown of XIST did have an i展开更多
Plasmacytoid dendritic cells(pDCs),also known as type I interferon(IFN)-producing cells,are specialized immune cells characterized by their extraordinary capabilities of mounting rapid and massive type I IFN response ...Plasmacytoid dendritic cells(pDCs),also known as type I interferon(IFN)-producing cells,are specialized immune cells characterized by their extraordinary capabilities of mounting rapid and massive type I IFN response to nu-cleic acids derived from virus,bacteria or dead cells.PDCs selectively express endosomal Toll-like receptor(TLR)7 and TLR9,which sense viral RNA and DNA re-spectively.Following type I IFN and cytokine responses,pDCs differentiate into antigen presenting cells and ac-quire the ability to regulate T cell-mediated adaptive immunity.The functions of pDCs have been implicated not only in antiviral innate immunity but also in immune tolerance,inflammation and tumor microenvironments.In this review,we will focus on TLR7/9 signaling and their regulation by pDC-specific receptors.展开更多
Phytohormones play crucial roles in fruit set regulation and development.Here,gibberellins(GA4+7),but not GA3,induced pear parthenocarpy.To systematically investigate the changes upon GA4+7 induced pear parthenocarpy,...Phytohormones play crucial roles in fruit set regulation and development.Here,gibberellins(GA4+7),but not GA3,induced pear parthenocarpy.To systematically investigate the changes upon GA4+7 induced pear parthenocarpy,dynamic changes in histology,hormone and transcript levels were observed and identified in unpollinated,pollinated and GA4+7-treated ovaries.Mesocarp cells continued developing in both GA4+7-treated and pollinated ovaries.In unpollinated ovaries,mesocarp cells stopped developing 14 days after anthesis.During fruit set process,GA4+7,but not GA1+3,increased after pollination.Abscisic acid(ABA)accumulation was significantly repressed by GA4+7 or pollination,but under unpollinated conditions,ABA was produced in large quantities.Moreover,indole-3-acetic acid biosynthesis was not induced by GA4+7 or pollination treatments.Details of this GA–auxin–ABA cross-linked gene network were determined by a comparative transcriptome analysis.The indole-3-acetic acid transport-related genes,mainly auxin efflux carrier component genes,were induced in both GA4+7-treated and pollinated ovaries.ABA biosynthetic genes of the 9-cis-epoxycarotenoid dioxygenase family were repressed by GA4+7 and pollination.Moreover,directly related genes in the downstream parthenocarpy network involved in cell division and expansion(upregulated),and MADS-box family genes(downregulated),were also identified.Thus,a model of GA-induced hormonal balance and its effects on parthenocarpy were established.展开更多
基金supported by a grant from the United States-Israel Binational Science Foundation(BSF 2007052,to W.J.L.and S.W.)by a Postdoctoral Fellowship for Research Abroad from the Japanese Society for the Promotion of Science(awarded to Michitaka Notaguchi).
文摘In plants, the phloem is the component of the vascular system that delivers nutrients and transmits signals from mature leaves to developing sink tissues. Recent studies have identified proteins, mRNA, and small RNA within the phloem sap of several plant species. It is now of considerable interest to elucidate the biological functions of these potential long-distance signal agents, to further our understanding of how plants coordinate their developmental programs at the whole-plant level. In this study, we developed a strategy for the functional analysis of phloem-mobile mRNA by focusing on IAA transcripts, whose mobility has previously been reported in melon （Cucumis melo cv. Hale＇s Best Jumbo）. Indoleacetic acid （IAA） proteins are key transcriptional regulators of auxin signaling, and are involved in a broad range of developmental processes including root development. We used a combination of vasculature-enriched sampling and hetero-grafting techniques to identify IAA18 and IAA28 as phloemmobile transcripts in the model plant Arabidopsis thaliana. Micro-grafting experiments were used to confirm that these IAA transcripts, which are generated in vascular tissues of mature leaves, are then transported into the root system where they negatively regulate lateral root formation. Based on these findings, we present a model in which auxin distribution, in combination with phloem-mobile Aux/IAA transcripts, can determine the sites of auxin action.
文摘Plant metabolomics developed as a powerful tool to examine gene functions and to gain deeper insight into the physiology of the plant cell. In this study, we screened Arabidopsis lines overexpressing rice full-length （FL） cDNAs （rice FOX Arabidopsis lines） using a gas chromatography-time-of-flight mass spectrometry （GC-TOF/MS）-based technique to identify rice genes that caused metabolic changes. This screening system allows fast and reliable identification of candi- date lines showing altered metabolite profiles. We performed metabolomic and transcriptomic analysis of a rice FOX Ara- bidopsis line that harbored the FL cDNA of the rice ortholog of the Lateral Organ Boundaries （LOB） Domain （LBD）/ Asymmetric Leaves2-1ike （ASL） gene of Arabidopsis, At-LBD37/ASL39. The investigated rice FOX Arabidopsis line showed prominent changes in the levels of metabolites related to nitrogen metabolism. The transcriptomic data as well as the results from the metabolite analysis of the Arabidopsis At-LBD37/ASL39-overexpressor plants were consistent with these findings. Furthermore, the metabolomic and transcriptomic analysis of the Os-LBD37/ASL39-overexpressing rice plants indicated that Os-LBD37/ASL39 is associated with processes related to nitrogen metabolism in rice. Thus, the combination of a metabolomics-based screening method and a gain-of-function approach is useful for rapid characterization of novel genes in both Arabidopsis and rice.
基金supported by the National Basic Research Program of China (No. 2010CB126100, 2009CB119006)the National Natural Science Foundation of China (No.21077093, 21277127)
文摘Silver nanoparticles （AgNPs） are one of the most widely used nanomaterials, but the mechanism of AgNP toxicity in terrestrial plants is still unclear. We compared the toxic effects of AgNPs and Ag＋ on Arabidopsis thaliana at the physiological, ultrastructural and molecular levels. AgNPs did not affect seed germination; however, they showed stronger inhibitory effect on root elongation than Ag＋ . The results of transmission electron microscopy and metal content analysis showed that AgNPs could be accumulated in leaves. These absorbed AgNPs disrupted the thylakoid membrane structure and decreased chlorophyll content, which can inhibit plant growth. By comparison, a small amount of Ag＋ was absorbed by seedlings, and it did not pronouncedly affect chloroplast structure and other metal ion absorption as AgNPs did. Compared with Ag＋ , AgNPs could alter the transcription of antioxidant and aquaporin genes, indicating that AgNPs changed the balance between the oxidant and antioxidant systems, and also affected the homeostasis of water and other small molecules within the plant body. All the data from physiological, ultrastructural and molecular levels suggest that AgNPs were more toxic than Ag＋ .
基金supported by the National Natural Science Foundation of China(Nos.81430072,81421003,81602641,81572929).
文摘Background:Gastric cancer(GC)is one of the most common malignancies worldwide,particularly in China.DNA damage-inducible transcript 4(DDIT4)is a mammalian target of rapamycin inhibitor and is induced by various cellular stresses;however,its critical role in GC remains poorly understood.The present study aimed to investigate the poten-tial relationship and the underlying mechanism between DDIT4 and GC development.Methods:We used western blotting,real-time polymerase chain reaction,and immunohistochemical or immunoflu-orescence to determine DDIT4 expression in GC cells and tissues.High-content screening,cell counting kit-8 assays,colony formation,and in vivo tumorigenesis assays were performed to evaluate cell proliferation.Flow cytometry was used to investigate cell apoptosis and cell cycle distribution.Results:DDIT4 was upregulated in GC cells and tissue.Furthermore,downregulating DDIT4 in GC cells inhibited proliferation both in vitro and in vivo and increased 5-fluorouracil-induced apoptosis and cell cycle arrest.In contrast,ectopic expression of DDIT4 in normal gastric epithelial cells promoted proliferation and attenuated chemosensitivity.Further analysis indicated that the mitogen-activated protein kinase and p53 signaling pathways were involved in the suppression of proliferation,and increased chemosensitivity upon DDIT4 downregulation.Conclusion:DDIT4 promotes GC proliferation and tumorigenesis,providing new insights into the role of DDIT4 in the tumorigenesis of human GC.
文摘The plasma membrane Na＋/H＋-antiporter salt overly sensitive1 （SOS1） from the halophytic Arabidopsis-relative Thellungiella halophila （ThSOS1） shows conserved sequence and domain structure with the orthologous genes from Arabidopsis thaliana and other plants. When expression of ThSOSt was reduced by RNA interference （RNAi）, pronounced characteristics of salt-sensitivity were observed. We were interested in monitoring altered transcriptional responses between Thellungiella wild type and thsost-4, a representative RNAi line with particular emphasis on root responses to salt stress at 350 mmol/L NaCI, a concentration that is only moderately stressful for mature wild type plants. Transcript profiling revealed several functional categories of genes that were differently affected in wild-type and RNAi plants. Down-regulation of SOS1 resulted in different gene expression even in the absence of stress. The pattern of gene induction in the RNAi plant under salt stress was similar to that of glycophytic Arabidopsis rather than that of wild type Thellungiella. The RNAi plants failed to down-regulate functions that are normally reduced in wild type Thellungiella upon stress and did not up-regulate functions that characterize the Thellungiella salt stress response. Metabolite changes observed in wild type Thellungiella after salt stress were less pronounced or absent in RNAi plants. Transcript and metabolite behavior suggested SOS1 functions including but also extending its established function as a sodium transporter. The down-regulation of ThSOS1 converted the halophyte Thellungiella into a salt-sensitive plant.
文摘Background Folic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of fotic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase （DHFR） gene knock-down zebrafish embryos, investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 （HAND2） and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations. Methods Morpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression. Results DHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos （P〈0.05）. Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos. Conclusions DHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation.
基金Supported by the Shandong Provincial Natural Science Foundation of China,No.ZR2016HQ08Shandong Province Medical Science and Technology Development Projects of China,No.2016WS0151the Jining Municipal Project on Science and Technology Development of China,No.2013jnwk58
文摘AIM To clarify the mechanisms of HOX transcript antisense intergenic RNA(HOTAIR) in gastric cancer(GC) migration and invasion.METHODS Quantitative real-time polymerase chain reaction(qp CR) was used to detect the expression level of HOTAIR in GC tissues. The correlation of its expression with clinicopathological features was analyzed. Area under receiver operating characteristic curve(AUCROC) was constructed to evaluate the diagnostic value of HOTAIR. Wound-healing assay and Transwell assay were performed to detect the biological effects of HOTAIR in GC cells. qp CR,western blot and immunohistochemistry were used to evaluate the m RNA and protein expression of E-cadherin. RNAbinding protein immunoprecipitation was used for the analysis of EZH2 interactions with HOTAIR. Chromatin immunoprecipitation assay was performed to investigate direct interactions between EZH2 and E-cadherin.RESULTS The expression of HOTAIR was up-regulated in GC tumorous tissues compared with the para-tumorous tissues(p < 0.001). Its over-expression was correlated with tumor-node-metastasis(TNM) stage(p = 0.024),tumor invasion(p = 0.018),lymph node metastasis(p = 0.023),and poor prognosis(p < 0.001). Multivariate Cox regression analysis confirmed expression of HOTAIR as an independent predictor of overall survival(p = 0.033),together with TNM stage(p = 0.002) and lymph node metastasis(p = 0.002). The AUCROC was up to 0.709(95%CI: 0.623-0.785,p < 0.001). Knockdown of HOTAIR by si RNA in GC cells suppressed the migration and invasion of GC cells. Significantly negative correlation between HOTAIR and E-cadherin was found in GC tissues and cell lines,and HOTAIR contributed to the regulation of E-cadherin through binding to EZH2 with the E-cadherin promoter. CONCLUSION HOTAIR may play a pivotal role in tumor cell migration and invasion. It can be used as a potential diagnostic and prognostic biomarker for GC.
基金Supported by the National Natural Science Foundation of China(No.81603441)Educational Commission of Sichuan Province(No.16ZA0282)the China Postdoctoral Science Foundation(No.2016M590872),China
文摘Objective： To study the expression level and role of apoptosis-associated speck-like protein containing a caspase recruitment domain （PYCARD） gene transcript variant mRNA in peripheral blood mononuclear cells （PBMCs） of primary gout （PG） patients with different Chinese medicine （CM） syndromes. Methods： The expressions of PYCARD gene transcript variant mRNA and interleukin-1β （IL-1β） mRNA in PBMCs were investigated in 96 PG patients with acute phase （APPG, 44 cases） and non-acute phase （NAPPG, 52 cases） and 30 healthy controls （HCs） by reverse transcription-polymerase chain reaction （PCR） and/or real-time quantitative PCR. PYCARD and nuclear factor-κB （p50） [NF-κB （p50）] protein was detected by Western blot in PBMCs respectively. IL-1β, IL-4 and IL-10 protein levels in plasma of HCs and PG patients were measured by enzyme-linked immuno sorbent assay. Results： The main CM syndromes in APPG patients were obstruction of dampness and heat syndrome （ODHS, 36.36%） and intermingled phlegm-blood stasis syndrome （IPBSS, 27.27%）, while in NAPPG patients were Pi （Spleen）-deficiency induced dampness syndrome （PDIDS, 40.38%） and qi-blood deficiency syndrome （QBDS, 26.92%）. It showed statistical significances of the expressions of PYCARD gene and its transcript variant mRNA, the protein of PYCARD and NF-κB （p50） and the plasma IL-1β, IL-4 and IL-10 in APPG, NAPPG, ODHS, IPBSS, PDIDS and QBDS groups, compared with the HC group respectively （P〈0.05 or P〈0.01）. There were also significant differences of mRNA expressions of PYCARD-1 and PYCARD-2 as well as protein expressions of IL-1β, IL-4 and IL-10 among the 4 CM syndromes groups （P〈0.05 or P〈0.01）. Correlation analysis showed positive correlation between the mRNA expressions of PYCARD-1 gene transcript variant and IL-1β in APPG patients （r=0.3088, P=0.0183）. Conclusion： PYCARD gene and its transcript variant may play a critical and regulative role in the inflammatory response
文摘BACKGROUND Anti-programmed death therapy has thrust immunotherapy into the spotlight.However,such therapy has a modest response in hepatocellular carcinoma(HCC).Epigenetic immunomodulation is a suggestive combinatorial therapy with immune checkpoint blockade.Non-coding ribonucleic acid(ncRNA)driven regulation is a major mechanism of epigenetic modulation.Given the wide range of ncRNAs that co-opt in programmed cell-death protein 1(PD-1)/programmed death ligand 1(PD-L1)regulation,and based on the literature,we hypothesized that miR-155-5p,miR-194-5p and long non-coding RNAs(lncRNAs)X-inactive specific transcript(XIST)and MALAT-1 are involved in a regulatory upstream pathway for PD-1/PD-L1.Recently,nutraceutical therapeutics in cancers have received increasing attention.Thus,it is interesting to study the impact of oleuropein on the respective study key players.AIM To explore potential upstream regulatory ncRNAs for the immune checkpoint PD-1/PD-L1.METHODS Bioinformatics tools including microrna.org and lnCeDB software were adopted to detect targeting of miR-155-5p,miR-194-5p and lncRNAs XIST and MALAT-1 to PD-L1 mRNA,respectively.In addition,Diana tool was used to predict targeting of both aforementioned miRNAs to lncRNAs XIST and MALAT-1.HCC and normal tissue samples were collected for scanning of PD-L1,XIST and MALAT-1 expression.To study the interaction among miR-155-5p,miR-194-5p,lncRNAs XIST and MALAT-1,as well as PD-L1 mRNA,a series of transfections of the Huh-7 cell line was carried out.RESULTS Bioinformatics software predicted that miR-155-5p and miR-194-5p can target PDL1,MALAT-1 and XIST.MALAT-1 and XIST were predicted to target PD-L1 mRNA.PD-L1 and XIST were significantly upregulated in 23 HCC biopsies compared to healthy controls;however,MALAT-1 was barely detected.MiR-194 induced expression elevated the expression of PD-L1,XIST and MALAT-1.However,overexpression of miR-155-5p induced the upregulation of PD-L1 and XIST,while it had a negative impact on MALAT-1 expression.Knockdown of XIST did have an i
文摘Plasmacytoid dendritic cells(pDCs),also known as type I interferon(IFN)-producing cells,are specialized immune cells characterized by their extraordinary capabilities of mounting rapid and massive type I IFN response to nu-cleic acids derived from virus,bacteria or dead cells.PDCs selectively express endosomal Toll-like receptor(TLR)7 and TLR9,which sense viral RNA and DNA re-spectively.Following type I IFN and cytokine responses,pDCs differentiate into antigen presenting cells and ac-quire the ability to regulate T cell-mediated adaptive immunity.The functions of pDCs have been implicated not only in antiviral innate immunity but also in immune tolerance,inflammation and tumor microenvironments.In this review,we will focus on TLR7/9 signaling and their regulation by pDC-specific receptors.
基金This work was supported by the China Agriculture Research System(CARS-29-40)Weinan Experimental Station foundation of Northwest A&F University.
文摘Phytohormones play crucial roles in fruit set regulation and development.Here,gibberellins(GA4+7),but not GA3,induced pear parthenocarpy.To systematically investigate the changes upon GA4+7 induced pear parthenocarpy,dynamic changes in histology,hormone and transcript levels were observed and identified in unpollinated,pollinated and GA4+7-treated ovaries.Mesocarp cells continued developing in both GA4+7-treated and pollinated ovaries.In unpollinated ovaries,mesocarp cells stopped developing 14 days after anthesis.During fruit set process,GA4+7,but not GA1+3,increased after pollination.Abscisic acid(ABA)accumulation was significantly repressed by GA4+7 or pollination,but under unpollinated conditions,ABA was produced in large quantities.Moreover,indole-3-acetic acid biosynthesis was not induced by GA4+7 or pollination treatments.Details of this GA–auxin–ABA cross-linked gene network were determined by a comparative transcriptome analysis.The indole-3-acetic acid transport-related genes,mainly auxin efflux carrier component genes,were induced in both GA4+7-treated and pollinated ovaries.ABA biosynthetic genes of the 9-cis-epoxycarotenoid dioxygenase family were repressed by GA4+7 and pollination.Moreover,directly related genes in the downstream parthenocarpy network involved in cell division and expansion(upregulated),and MADS-box family genes(downregulated),were also identified.Thus,a model of GA-induced hormonal balance and its effects on parthenocarpy were established.
