SUPPRESSOR OF OVEREXPRESSION OF CO 1(SOC1)编码MADS-box转录因子,具有诱导植物成花、防止花分生组织过早成熟和调控花器官发育的重要功能,是调控植物开花的整合因子。SOC1转录因子受蛋白或核酸调控,或与其他转录因子形成复合物进入...SUPPRESSOR OF OVEREXPRESSION OF CO 1(SOC1)编码MADS-box转录因子,具有诱导植物成花、防止花分生组织过早成熟和调控花器官发育的重要功能,是调控植物开花的整合因子。SOC1转录因子受蛋白或核酸调控,或与其他转录因子形成复合物进入细胞核,靶向特异开花基因从而调控植物开花,其调控网络十分复杂。本研究主要分析SOC1及其编码蛋白的结构特征,预测SOC1的蛋白互作网络,并阐明SOC1调控植物开花时间的分子机制,SOC1的表达受FT、CO、FLC及其同源基因MAFs、SVP、SPL和AGL24的直接调控,并受DELLA、miR156、miR172、AP2类转录因子、MYC3蛋白及营养物质信号的间接调控,且与AGL24形成二聚体上调下游花分生组织基因LFY的表达来调控植物的开花诱导过程,为今后研究SOC1调控其他植物的开花时间提供参考。展开更多
AIM:To investigate the stability of the seven housekeeping genes:beta-actin(ActB),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),18s ribosomal unit 5(18s),cyclophilin A(CycA),hypoxanthine-guanine phosphoribosyl trans...AIM:To investigate the stability of the seven housekeeping genes:beta-actin(ActB),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),18s ribosomal unit 5(18s),cyclophilin A(CycA),hypoxanthine-guanine phosphoribosyl transferase(HPRT),ribosomal protein large P0(36B4)and terminal uridylyl transferase 1(U6)in the diabetic retinal tissue of rat model.METHODS:The expression of these seven genes in rat retinal tissues was determined using real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)in two groups;normal control rats and streptozotocininduced diabetic rats.The stability analysis of gene expression was investigated using geNorm,NormFinder,BestKeeper,and comparative delta-Ct(ΔCt)algorithms.RESULTS:The 36B4 gene was stably expressed in the retinal tissues of normal control animals;however,it was less stable in diabetic retinas.The 18s gene was expressed consistently in both normal control and diabetic rats’retinal tissue.That this gene was the best reference for data normalisation in RT-qPCR studies that used the retinal tissue of streptozotocin-induced diabetic rats.Furthermore,there was no ideal gene stably expressed for use in all experimental settings.CONCLUSION:Identifying relevant genes is a need for achieving RT-qPCR validity and reliability and must be appropriately achieved based on a specific experimental setting.展开更多
In eukaryotes,MEDIATOR is a conserved multisubunit complex that links transcription factors and RNA polymerase II and that thereby facilitates transcriptional initiation.Although the composition of MEDIATOR has been w...In eukaryotes,MEDIATOR is a conserved multisubunit complex that links transcription factors and RNA polymerase II and that thereby facilitates transcriptional initiation.Although the composition of MEDIATOR has been well studied in yeast and mammals,relatively little is known about the composition of MEDIATOR in plants.By affinity purification followed by mass spectrometry,we identified 28 conserved MEDIATOR subunits in Arabidopsis thaliana,including putative MEDIATOR subunits that were not previously validated.Our results indicated that MED34,MED35,MED36,and MED37 are not Arabidopsis MEDIATOR subunits,as previously proposed.Our results also revealed that two homologous CBP/p300 histone acetyltransferases,HAC1 and HAC5(HAC1/5)are in fact plant-specific MEDIATOR subunits.The MEDIATOR subunits MED8 and MED25(MED8/25)are partially responsible for the association of MEDIATOR with HAC1/5,MED8/25 and HAC1/5 co-regulate gene expression and thereby affect flowering time and floral development.Our in vitro observations indicated that MED8 and HAC1 form liquid-like droplets by phase separation,and our in vivo observations indicated that these droplets co-localize in the nuclear bodies at a subset of nuclei.The formation of liquid-like droplets is required for MED8 to interact with RNA polymerase II.In summary,we have identified all of the components of Arabidopsis MEDIATOR and revealed the mechanism underlying the link of histone acetylation and transcriptional regulation.展开更多
The mature seed of Paris polyphylla var.chinensis(PPC)is morphophysiologically dormant and develops differently under warm and cold temperatures.To elucidate the molecular mechanisms underlying temperature-dependent r...The mature seed of Paris polyphylla var.chinensis(PPC)is morphophysiologically dormant and develops differently under warm and cold temperatures.To elucidate the molecular mechanisms underlying temperature-dependent regulation of PPC seed dormancy and germination,we investigated the dynamic changes in PPC seed transcript levels under warm and cold temperature stratifications(WS and CS,respectively)by time-resolved RNA sequencing,focusing on genes related to hormone metabolism and signaling and cell wall remodeling(CWRM)and encoding transcription factors/regulators(TFs/TRs).A total of 48765 and 47836 differentially expressed genes(DEGs)were associated with WS and CS,respectively.Of these,17581 and 16652 DEGs,respectively,unique to WS and CS,and 5386 were common to both temperature stratifications across nine analyzed growth stages.The expression of hormone metabolism and signaling,TF/TR,and CWRM genes were closely associated with temperature.More genes related to gibberellin(GA),cytokinin,auxin,and brassinosteroid biosynthetic were upregulated in WS as compared to CS seeds,while genes related to dormancy release and germination were downregulated in WS seeds.However,only GA and abscisic acid levels were altered in PPC seeds breaking morphophysiological dormancy(MPD).Overall,37 TF and five TR families were upregulated whereas 24 TF and 16 TR families were downregulated in WS as compared to CS seeds.Most CWRM families were highly expressed under WS as compared to CS,suggesting that they promote endosperm weakening and embryo growth of WS seeds and facilitate MPD release and germination.A coexpression analysis revealed positive correlations between TFs/TRs and DEGs involved in hormone metabolism and signaling and CWRM.These results provided a basis for investigating the interaction between the endosperm and underdeveloped embryo in the regulation of PPC seed germination and seedling emergence.展开更多
Monocarpic senescence,characterized by whole-plant senescence following a single flowering phase,is widespread in seed plants,particularly in crops,determining seed harvest time and quality.However,how external and in...Monocarpic senescence,characterized by whole-plant senescence following a single flowering phase,is widespread in seed plants,particularly in crops,determining seed harvest time and quality.However,how external and internal signals are systemically integrated into monocarpic senescence remains largely unknown.Here,we report that the Arabidopsis thaliana transcription factor WRKY1 plays essential roles in multiple key steps of monocarpic senescence.WRKY1 expression is induced by age,salicylic acid(SA),and nitrogen(N)deficiency.Flowering and leaf senescence are accelerated in the WRKY1 overexpression lines but are delayed in the wrky1 mutants.The combined DNA affinity purification sequencing and RNA sequencing analyses uncover the direct target genes of WRKY1.Further studies show that WRKY1 coordinately regulates three processes in monocarpic senescence:(1)suppressing FLOWERING LOCUS C gene expression to initiate flowering,(2)inducing SA biosynthesis genes to promote leaf senescence,and(3)activating the N assimilation and transport genes to trigger N remobilization.In summary,our study reveals how one stress-responsive transcription factor,WRKY1,integrates flowering,leaf senescence,and N remobilization processes into monocarpic senescence,providing important insights into plant lifetime regulation.展开更多
The purpose of this study was to screen key survival-related genes from patients with colorectal cancer and explore signal transduction network of the involved genes.In a previous study,survival-related genes of patie...The purpose of this study was to screen key survival-related genes from patients with colorectal cancer and explore signal transduction network of the involved genes.In a previous study,survival-related genes of patients with colorectal cancer were selected by colorectal cancer-related expression data GSE17538 using the Significance Analysis of Microarrays(SAM3.01)software,and 235 genes related to the survival of patients with colorectal cancer were obtained.Therefore,the following screening and analysis were conducted on these 235 genes in this study.First,the enrichment analysis of transcription factor binding sites was conducted on the 235 genes.Genes with more than seven transcription factor binding sites were screened.Then,these genes and upregulated genes in colorectal cancer were intersected.Finally,survival analysis and regulatory network analysis were conducted on the screened genes.This allowed clarification of the relationship between these genes and the survival of patients with colorectal cancer and the signaling network involving these genes in the cell signal transduction network of colorectal cancer.Through the above analysis,six upregulated genes in colorectal cancer related to the survival of colorectal cancer patients and highly regulated by transcription factors were selected,namely STX2,PODXL,KLK6,GRB10,EHBP1 and CREB5.These genes are involved in signal regulatory networks related to colorectal cancer metastasis-related signaling pathways.Therefore,the survival of patients with colorectal cancer is closely correlated with colorectal cancer metastasis.The six survival-related genes affect the survival of patients by regulating colorectal cancer metastasis-associated signaling pathways.展开更多
Objective: The aim of this study was to determine the etiology and prevalence of pediatric CAP in Beijing using a real-time polymerase chain reaction (PCR) technique. Methods: Between February 15, 2011 and January 18...Objective: The aim of this study was to determine the etiology and prevalence of pediatric CAP in Beijing using a real-time polymerase chain reaction (PCR) technique. Methods: Between February 15, 2011 and January 18, 2012, 371 pediatric patients with CAP were enrolled at Beijing Children's Hospital. Sixteen respiratory viruses and two bacteria were detected from tracheal aspirate specimens using commercially available multiplex real-time reverse transcription PCR (RT-PCR) kits. Results: A single viral pathogen was detected in 35.3%of enrolled patients, multiple viruses in 11.6%, and virus/bacteria co-infection in 17.8%. In contrast, only 6.5%of patients had a single bacterial pathogen and 2.2%were infected with multiple bacteria. The etiological agent was unknown for 26.7% of patients. The most common viruses were respiratory syncytial virus (RSV) (43.9%), rhinovirus (14.8%), parainfluenza virus (9.4%), and adenovirus (8.6%). In patients under three years of age, RSV (44.6%), rhinovirus (12.8%), and Streptococcus pneumoniae (9.9%) were the most frequent pathogens. In children aged 3e7 years, S. pneumoniae (38.9%), RSV (30.6%), Haemophilus influenzae (19.4%), and adenovirus (19.4%) were most prevalent. Finally in children over seven years, RSV (47.3%), S. pneumoniae (41.9%), and rhinovirus (21.5%) infections were most frequent. Conclusions: Viral pathogens, specifically RSV, were responsible for the majority of CAP in pediatric patients. However, both S. pneumoniae and H. influenzae contributed as major causes of disease. Commercially available multiplexing real-time PCR allowed for rapid detection of the etiological agent. Copyright ? 2015, Chinese Medical Association Production. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).展开更多
Flowering time is crucial for successful reproduction in plants, the onset and progression of which are strictly controlled. However, flowering time is a complex and environmentally responsive history trait and the un...Flowering time is crucial for successful reproduction in plants, the onset and progression of which are strictly controlled. However, flowering time is a complex and environmentally responsive history trait and the underlying mechanisms still need to be fully characterized. Post-translational regulation of the activities of transcription factors(TFs) is a dynamic and essential mechanism for plant growth and development. CRL3 BPME3 ligase is a CULLIN3-based E3 ligase involved in orchestrating protein stability via the ubiquitin proteasome pathway. Our study shows that the mutation of MYB106 induced early flowering phenotype while over-expression of MYB106 delayed Arabidopsis flowering. Transcriptome analysis of myb106 mutants reveals 257 differentially expressed genes between wild type and myb106-1 mutants, including Flowering Locus T(FT) which is related to flowering time. Moreover, in vitro electrophoretic mobility shift assays(EMSA), in vivo chromatin immunoprecipitation quantitative polymerase chain reaction(ChIP-q PCR) assays and dual luciferase assays demonstrate that MYB106 directly binds to the promoter of FT to suppress its expression. Furthermore, we confirm that MYB106 interacts with BPM proteins which are further identified by CRL3 BPME3 ligases as the substrate. Taken together, we have identified MYB106 as a negative regulator in the control of flowering time and a new substrate for CRL3 BPM E3 ligases in Arabidopsis.展开更多
A quantitative real time reverse-transcription polymerase chain reaction （qRT-PCR） assay with specific primers recommended by the World Health Organization （WHO） has been widely used successfully for detection and...A quantitative real time reverse-transcription polymerase chain reaction （qRT-PCR） assay with specific primers recommended by the World Health Organization （WHO） has been widely used successfully for detection and monitoring of the pandemic H1N1/2009 influenza A virus. In this study, we report the design and characterization of a novel set of primers to be used in a qRT-PCR assay for detecting the pandemic H1N1/2009 virus. The newly designed primers target three regions that are highly conserved among the hemagglutinin （HA） genes of the pandemic HlN1/2009 viruses and are different from those targeted by the WHO-recommended primers. The qRT-PCR assays with the newly designed primers are highly specific, and as specific as the WHO-recommended primers for detecting pandemic H1N1/2009 viruses and other influenza viruses including influenza B viruses and influenza A viruses of human, swine, and raccoon dog origin. Furthermore, the qRT-PCR assays with the newly designed primers appeared to be at least 10-fold more sensitive than those with the WHO-recommended primers as the detection limits of the assays with our primers and the WHO-recommended primers were 2.5 and 25 copies of target RNA per reaction, respectively. When tested with 83 clinical samples, 32 were detected to be positive using the qRT-PCR assays with our designed primers, while only 25 were positive by the assays with the WHO-recommended primers. These results suggest that the qRT-PCR system with the newly designed primers represent a highly sensitive assay for diagnosis of the pandemic HIN1/2009 virus infection.展开更多
BACKGROUND The global outbreak of coronavirus disease 2019(COVID-19)leads to the development of accessible and cost-effective rapid antigen-detection tests(RATs),as quick and accurate diagnosis is crucial to curb the ...BACKGROUND The global outbreak of coronavirus disease 2019(COVID-19)leads to the development of accessible and cost-effective rapid antigen-detection tests(RATs),as quick and accurate diagnosis is crucial to curb the pandemic.AIM To evaluate the Humasis COVID-19 Ag Test(Humasis Co.,Ltd.,Gyeonggi-do,Republic of Korea)in the diagnosis of severe acute respiratory syndrome coronavirus-2(SARS-CoV-2).METHODS This retrospective study was carried out at the Croatian Institute of Public Health and included patients with clinical symptoms of COVID-19 lasting no longer than 5 d prior to testing,whose nasopharyngeal swabs were primarily tested with RAT.Negative RAT samples underwent confirmatory real-time reverse transcription-polymerase chain reaction(RT-PCR).Diagnostic efficacy was determined compared to RT-PCR.The patients were divided into three age groups(<18,19-65,>65 years).Statistical analysis was performed with the significance level set at P<0.05.RESULTSIn total,2490 symptomatic patients were tested;953 samples were positive on RAT,and 1537 werenegative.All negative RAT samples were subjected to RT-PCR;266 samples were positive andmarked as false-negative results on RAT.The calculated negative predictive value as a measure ofRAT efficacy was 82.69%.The χ^(2) test and Kruskal-Wallis test showed a significant difference in theproportion of false negatives(P<0.001)and RT-PCR cycle(Ct)values for false-negative RATs(P=0.012)among the age groups.The young age group was significantly less likely to be falsenegative,whereas the false negatives from the elderly group experienced significantly lower Ctvalues than the other two age groups.CONCLUSIONEvaluated RAT demonstrated satisfactory performance with more reliable results in youngerpatients.Humasis COVID-19 Ag RAT is potentially a valuable tool in areas where access tomolecular methods is limited;however,RT-PCR remains a gold standard for SARS-CoV-2detection.展开更多
The technique of SYBR Green-based quantitative real-time reverse transcription polymerase chain reaction(real-time RT-PCR)was applied to quantitative detect a 764 bp nucleotide sequence containing total coat protein(c...The technique of SYBR Green-based quantitative real-time reverse transcription polymerase chain reaction(real-time RT-PCR)was applied to quantitative detect a 764 bp nucleotide sequence containing total coat protein(cp)gene of Cymbidium mosaic virus(CyMV).The plasmid containing the target sequence was constructed to prepare the standard curve and detect the sensitivity.The standard curve was drawn based on the linear relationship between the logarithm(base 10)of the quantity of target sequence and cycle threshold[C(T)].While the concentration of plasmid DNA falling within the range of 2.6×10^(7)to 2.6×10^(2)copies per tube established a regression equation,y=-0.3583x+10.32,and related coefficient:r^(2)=0.995.The real-time RT-PCR assay for CyMV had a minimum detectable quantity of two copies per tube.The naturally infected samples of Phalaenopsis sp.and the artificially inoculated samples of Arachnis sp.with trace CyMV were quantitatively detected using this method.CyMVin the positive samples of Phalaenopsis sp.and Arachnis sp.was confirmed by DNA sequencing and cp gene homeology blast.The results showed that CyMV extracted from the leaves of orchid in Hangzhou,Zhejiang Province,China,could be derived from Kunming city(KM),Yunnan Province,China.This method characterized by high sensitivity,specificity,and precision is suitable for early diagnosis and quantitative detection of CyMV.展开更多
Defining con-founders that affect the reliability of diagnostic tests for coronavirus disease 2019 is vital to breaking the chain of infection.The elderly population is a higher risk group for the emerging virus.Howev...Defining con-founders that affect the reliability of diagnostic tests for coronavirus disease 2019 is vital to breaking the chain of infection.The elderly population is a higher risk group for the emerging virus.However,gender seems to exert a critical role in modifying the infection risk among women owing to hormonal changes.The menopause transition is an exceptional period for older women where the protective and immunomodulatory effects of the estrogen hormone are lost.Accordingly,attention should be given to postmenopausal women since they will have an increased risk compared to their pre-menopausal peers.展开更多
The B-cell lymphoma/leukemia 11A （BCL11A） gene is essential for normal lymphoid development and has been associated with hematological malignancies. In the current study, the relative expression level of BCL11A in m...The B-cell lymphoma/leukemia 11A （BCL11A） gene is essential for normal lymphoid development and has been associated with hematological malignancies. In the current study, the relative expression level of BCL11A in malignant hematological cell lines was evaluated through real-time quantitative reverse transcription polymerase chain reaction （qRT-PCR）. METHODS The relative expression level of BCLllA mRNA in malignant hematological cell lines was determined through qRT- PCR using SYBR Green I dye. Glyceraldehyde-3-phosphate dehydro- genase was used as the reference gene to confirm the relative expression level of BCL11A gene mRNA. RESULTS The relative expression level of BCL11A mRNA in cell lines from B-cell malignancies was significantly higher compared with that from acute rnyeloid leukemia （P 〈 0.05）. Different cell lines with malignant B-cells exhibited a wide range of BCL11A expressions ranging from 27.37 to 93.38. CONCLUSION The overexpression of BCL11A gene mRNA in malignant B-cells might play a role in B-cell lymphoma/leukemia.展开更多
文摘SUPPRESSOR OF OVEREXPRESSION OF CO 1(SOC1)编码MADS-box转录因子,具有诱导植物成花、防止花分生组织过早成熟和调控花器官发育的重要功能,是调控植物开花的整合因子。SOC1转录因子受蛋白或核酸调控,或与其他转录因子形成复合物进入细胞核,靶向特异开花基因从而调控植物开花,其调控网络十分复杂。本研究主要分析SOC1及其编码蛋白的结构特征,预测SOC1的蛋白互作网络,并阐明SOC1调控植物开花时间的分子机制,SOC1的表达受FT、CO、FLC及其同源基因MAFs、SVP、SPL和AGL24的直接调控,并受DELLA、miR156、miR172、AP2类转录因子、MYC3蛋白及营养物质信号的间接调控,且与AGL24形成二聚体上调下游花分生组织基因LFY的表达来调控植物的开花诱导过程,为今后研究SOC1调控其他植物的开花时间提供参考。
基金Supported by grant from Fundamental Research Grant Scheme by Ministry of Higher Education(MoHE)600-IRMI/FRGS 5/3(101/2019).
文摘AIM:To investigate the stability of the seven housekeeping genes:beta-actin(ActB),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),18s ribosomal unit 5(18s),cyclophilin A(CycA),hypoxanthine-guanine phosphoribosyl transferase(HPRT),ribosomal protein large P0(36B4)and terminal uridylyl transferase 1(U6)in the diabetic retinal tissue of rat model.METHODS:The expression of these seven genes in rat retinal tissues was determined using real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)in two groups;normal control rats and streptozotocininduced diabetic rats.The stability analysis of gene expression was investigated using geNorm,NormFinder,BestKeeper,and comparative delta-Ct(ΔCt)algorithms.RESULTS:The 36B4 gene was stably expressed in the retinal tissues of normal control animals;however,it was less stable in diabetic retinas.The 18s gene was expressed consistently in both normal control and diabetic rats’retinal tissue.That this gene was the best reference for data normalisation in RT-qPCR studies that used the retinal tissue of streptozotocin-induced diabetic rats.Furthermore,there was no ideal gene stably expressed for use in all experimental settings.CONCLUSION:Identifying relevant genes is a need for achieving RT-qPCR validity and reliability and must be appropriately achieved based on a specific experimental setting.
基金supported by the National Natural Science Foundation of China(32025003)by the National Key Research and Development Program of China(2016YFA0500801)from the Chinese Ministry of Science and Technology。
文摘In eukaryotes,MEDIATOR is a conserved multisubunit complex that links transcription factors and RNA polymerase II and that thereby facilitates transcriptional initiation.Although the composition of MEDIATOR has been well studied in yeast and mammals,relatively little is known about the composition of MEDIATOR in plants.By affinity purification followed by mass spectrometry,we identified 28 conserved MEDIATOR subunits in Arabidopsis thaliana,including putative MEDIATOR subunits that were not previously validated.Our results indicated that MED34,MED35,MED36,and MED37 are not Arabidopsis MEDIATOR subunits,as previously proposed.Our results also revealed that two homologous CBP/p300 histone acetyltransferases,HAC1 and HAC5(HAC1/5)are in fact plant-specific MEDIATOR subunits.The MEDIATOR subunits MED8 and MED25(MED8/25)are partially responsible for the association of MEDIATOR with HAC1/5,MED8/25 and HAC1/5 co-regulate gene expression and thereby affect flowering time and floral development.Our in vitro observations indicated that MED8 and HAC1 form liquid-like droplets by phase separation,and our in vivo observations indicated that these droplets co-localize in the nuclear bodies at a subset of nuclei.The formation of liquid-like droplets is required for MED8 to interact with RNA polymerase II.In summary,we have identified all of the components of Arabidopsis MEDIATOR and revealed the mechanism underlying the link of histone acetylation and transcriptional regulation.
基金supported by the CAMS Innovation Fund for Medical Sciences (CIFMS) (2017-I2M-3-013)the National Key Research and Development Program of China (Grant No. 2017YFC1700706)the National Natural Science Foundation of China (Grant No. 31471575).
文摘The mature seed of Paris polyphylla var.chinensis(PPC)is morphophysiologically dormant and develops differently under warm and cold temperatures.To elucidate the molecular mechanisms underlying temperature-dependent regulation of PPC seed dormancy and germination,we investigated the dynamic changes in PPC seed transcript levels under warm and cold temperature stratifications(WS and CS,respectively)by time-resolved RNA sequencing,focusing on genes related to hormone metabolism and signaling and cell wall remodeling(CWRM)and encoding transcription factors/regulators(TFs/TRs).A total of 48765 and 47836 differentially expressed genes(DEGs)were associated with WS and CS,respectively.Of these,17581 and 16652 DEGs,respectively,unique to WS and CS,and 5386 were common to both temperature stratifications across nine analyzed growth stages.The expression of hormone metabolism and signaling,TF/TR,and CWRM genes were closely associated with temperature.More genes related to gibberellin(GA),cytokinin,auxin,and brassinosteroid biosynthetic were upregulated in WS as compared to CS seeds,while genes related to dormancy release and germination were downregulated in WS seeds.However,only GA and abscisic acid levels were altered in PPC seeds breaking morphophysiological dormancy(MPD).Overall,37 TF and five TR families were upregulated whereas 24 TF and 16 TR families were downregulated in WS as compared to CS seeds.Most CWRM families were highly expressed under WS as compared to CS,suggesting that they promote endosperm weakening and embryo growth of WS seeds and facilitate MPD release and germination.A coexpression analysis revealed positive correlations between TFs/TRs and DEGs involved in hormone metabolism and signaling and CWRM.These results provided a basis for investigating the interaction between the endosperm and underdeveloped embryo in the regulation of PPC seed germination and seedling emergence.
基金the Zhejiang Provincial Natural Science Foundation of China(grant LZ23C020001 to K.Z.)the National Natural Science Foundation of China(grant 31670277 to K.Z.).
文摘Monocarpic senescence,characterized by whole-plant senescence following a single flowering phase,is widespread in seed plants,particularly in crops,determining seed harvest time and quality.However,how external and internal signals are systemically integrated into monocarpic senescence remains largely unknown.Here,we report that the Arabidopsis thaliana transcription factor WRKY1 plays essential roles in multiple key steps of monocarpic senescence.WRKY1 expression is induced by age,salicylic acid(SA),and nitrogen(N)deficiency.Flowering and leaf senescence are accelerated in the WRKY1 overexpression lines but are delayed in the wrky1 mutants.The combined DNA affinity purification sequencing and RNA sequencing analyses uncover the direct target genes of WRKY1.Further studies show that WRKY1 coordinately regulates three processes in monocarpic senescence:(1)suppressing FLOWERING LOCUS C gene expression to initiate flowering,(2)inducing SA biosynthesis genes to promote leaf senescence,and(3)activating the N assimilation and transport genes to trigger N remobilization.In summary,our study reveals how one stress-responsive transcription factor,WRKY1,integrates flowering,leaf senescence,and N remobilization processes into monocarpic senescence,providing important insights into plant lifetime regulation.
基金supported by the Joint Funds of the National Natural Science Foundation of China(U1201226)the National Natural Science Foundation of China(30670967)
文摘The purpose of this study was to screen key survival-related genes from patients with colorectal cancer and explore signal transduction network of the involved genes.In a previous study,survival-related genes of patients with colorectal cancer were selected by colorectal cancer-related expression data GSE17538 using the Significance Analysis of Microarrays(SAM3.01)software,and 235 genes related to the survival of patients with colorectal cancer were obtained.Therefore,the following screening and analysis were conducted on these 235 genes in this study.First,the enrichment analysis of transcription factor binding sites was conducted on the 235 genes.Genes with more than seven transcription factor binding sites were screened.Then,these genes and upregulated genes in colorectal cancer were intersected.Finally,survival analysis and regulatory network analysis were conducted on the screened genes.This allowed clarification of the relationship between these genes and the survival of patients with colorectal cancer and the signaling network involving these genes in the cell signal transduction network of colorectal cancer.Through the above analysis,six upregulated genes in colorectal cancer related to the survival of colorectal cancer patients and highly regulated by transcription factors were selected,namely STX2,PODXL,KLK6,GRB10,EHBP1 and CREB5.These genes are involved in signal regulatory networks related to colorectal cancer metastasis-related signaling pathways.Therefore,the survival of patients with colorectal cancer is closely correlated with colorectal cancer metastasis.The six survival-related genes affect the survival of patients by regulating colorectal cancer metastasis-associated signaling pathways.
文摘Objective: The aim of this study was to determine the etiology and prevalence of pediatric CAP in Beijing using a real-time polymerase chain reaction (PCR) technique. Methods: Between February 15, 2011 and January 18, 2012, 371 pediatric patients with CAP were enrolled at Beijing Children's Hospital. Sixteen respiratory viruses and two bacteria were detected from tracheal aspirate specimens using commercially available multiplex real-time reverse transcription PCR (RT-PCR) kits. Results: A single viral pathogen was detected in 35.3%of enrolled patients, multiple viruses in 11.6%, and virus/bacteria co-infection in 17.8%. In contrast, only 6.5%of patients had a single bacterial pathogen and 2.2%were infected with multiple bacteria. The etiological agent was unknown for 26.7% of patients. The most common viruses were respiratory syncytial virus (RSV) (43.9%), rhinovirus (14.8%), parainfluenza virus (9.4%), and adenovirus (8.6%). In patients under three years of age, RSV (44.6%), rhinovirus (12.8%), and Streptococcus pneumoniae (9.9%) were the most frequent pathogens. In children aged 3e7 years, S. pneumoniae (38.9%), RSV (30.6%), Haemophilus influenzae (19.4%), and adenovirus (19.4%) were most prevalent. Finally in children over seven years, RSV (47.3%), S. pneumoniae (41.9%), and rhinovirus (21.5%) infections were most frequent. Conclusions: Viral pathogens, specifically RSV, were responsible for the majority of CAP in pediatric patients. However, both S. pneumoniae and H. influenzae contributed as major causes of disease. Commercially available multiplexing real-time PCR allowed for rapid detection of the etiological agent. Copyright ? 2015, Chinese Medical Association Production. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
基金supported by National Natural Science Foundation of China(31670179,and 91854201)the Research Grants Council of Hong Kong(Ao E/M-05/12,CUHK14104716,and C4002-17G)to L.J.+1 种基金RGC(CUHK14104716)CUHK Direct Grants(4053143,4053174,4053243)to L.C.
文摘Flowering time is crucial for successful reproduction in plants, the onset and progression of which are strictly controlled. However, flowering time is a complex and environmentally responsive history trait and the underlying mechanisms still need to be fully characterized. Post-translational regulation of the activities of transcription factors(TFs) is a dynamic and essential mechanism for plant growth and development. CRL3 BPME3 ligase is a CULLIN3-based E3 ligase involved in orchestrating protein stability via the ubiquitin proteasome pathway. Our study shows that the mutation of MYB106 induced early flowering phenotype while over-expression of MYB106 delayed Arabidopsis flowering. Transcriptome analysis of myb106 mutants reveals 257 differentially expressed genes between wild type and myb106-1 mutants, including Flowering Locus T(FT) which is related to flowering time. Moreover, in vitro electrophoretic mobility shift assays(EMSA), in vivo chromatin immunoprecipitation quantitative polymerase chain reaction(ChIP-q PCR) assays and dual luciferase assays demonstrate that MYB106 directly binds to the promoter of FT to suppress its expression. Furthermore, we confirm that MYB106 interacts with BPM proteins which are further identified by CRL3 BPME3 ligases as the substrate. Taken together, we have identified MYB106 as a negative regulator in the control of flowering time and a new substrate for CRL3 BPM E3 ligases in Arabidopsis.
基金supported by grants from National Basic Research Program of China (No.2011CB504800)National Natural Science Foundation of China (No. 31100128 and 81030031)+3 种基金National Mega Project on Major Drug Development (2009ZX09103-678)National Small Business Innovation and Research (SBIR) Program of Chinathe Technology R & D Program of Jiangsu Province, China (BG20077035 and BG2008662)NIH (RO1-AI041927,RO1-AI050468, RO1-DE014145, and RO1-DE014842)
文摘A quantitative real time reverse-transcription polymerase chain reaction （qRT-PCR） assay with specific primers recommended by the World Health Organization （WHO） has been widely used successfully for detection and monitoring of the pandemic H1N1/2009 influenza A virus. In this study, we report the design and characterization of a novel set of primers to be used in a qRT-PCR assay for detecting the pandemic H1N1/2009 virus. The newly designed primers target three regions that are highly conserved among the hemagglutinin （HA） genes of the pandemic HlN1/2009 viruses and are different from those targeted by the WHO-recommended primers. The qRT-PCR assays with the newly designed primers are highly specific, and as specific as the WHO-recommended primers for detecting pandemic H1N1/2009 viruses and other influenza viruses including influenza B viruses and influenza A viruses of human, swine, and raccoon dog origin. Furthermore, the qRT-PCR assays with the newly designed primers appeared to be at least 10-fold more sensitive than those with the WHO-recommended primers as the detection limits of the assays with our primers and the WHO-recommended primers were 2.5 and 25 copies of target RNA per reaction, respectively. When tested with 83 clinical samples, 32 were detected to be positive using the qRT-PCR assays with our designed primers, while only 25 were positive by the assays with the WHO-recommended primers. These results suggest that the qRT-PCR system with the newly designed primers represent a highly sensitive assay for diagnosis of the pandemic HIN1/2009 virus infection.
文摘BACKGROUND The global outbreak of coronavirus disease 2019(COVID-19)leads to the development of accessible and cost-effective rapid antigen-detection tests(RATs),as quick and accurate diagnosis is crucial to curb the pandemic.AIM To evaluate the Humasis COVID-19 Ag Test(Humasis Co.,Ltd.,Gyeonggi-do,Republic of Korea)in the diagnosis of severe acute respiratory syndrome coronavirus-2(SARS-CoV-2).METHODS This retrospective study was carried out at the Croatian Institute of Public Health and included patients with clinical symptoms of COVID-19 lasting no longer than 5 d prior to testing,whose nasopharyngeal swabs were primarily tested with RAT.Negative RAT samples underwent confirmatory real-time reverse transcription-polymerase chain reaction(RT-PCR).Diagnostic efficacy was determined compared to RT-PCR.The patients were divided into three age groups(<18,19-65,>65 years).Statistical analysis was performed with the significance level set at P<0.05.RESULTSIn total,2490 symptomatic patients were tested;953 samples were positive on RAT,and 1537 werenegative.All negative RAT samples were subjected to RT-PCR;266 samples were positive andmarked as false-negative results on RAT.The calculated negative predictive value as a measure ofRAT efficacy was 82.69%.The χ^(2) test and Kruskal-Wallis test showed a significant difference in theproportion of false negatives(P<0.001)and RT-PCR cycle(Ct)values for false-negative RATs(P=0.012)among the age groups.The young age group was significantly less likely to be falsenegative,whereas the false negatives from the elderly group experienced significantly lower Ctvalues than the other two age groups.CONCLUSIONEvaluated RAT demonstrated satisfactory performance with more reliable results in youngerpatients.Humasis COVID-19 Ag RAT is potentially a valuable tool in areas where access tomolecular methods is limited;however,RT-PCR remains a gold standard for SARS-CoV-2detection.
文摘The technique of SYBR Green-based quantitative real-time reverse transcription polymerase chain reaction(real-time RT-PCR)was applied to quantitative detect a 764 bp nucleotide sequence containing total coat protein(cp)gene of Cymbidium mosaic virus(CyMV).The plasmid containing the target sequence was constructed to prepare the standard curve and detect the sensitivity.The standard curve was drawn based on the linear relationship between the logarithm(base 10)of the quantity of target sequence and cycle threshold[C(T)].While the concentration of plasmid DNA falling within the range of 2.6×10^(7)to 2.6×10^(2)copies per tube established a regression equation,y=-0.3583x+10.32,and related coefficient:r^(2)=0.995.The real-time RT-PCR assay for CyMV had a minimum detectable quantity of two copies per tube.The naturally infected samples of Phalaenopsis sp.and the artificially inoculated samples of Arachnis sp.with trace CyMV were quantitatively detected using this method.CyMVin the positive samples of Phalaenopsis sp.and Arachnis sp.was confirmed by DNA sequencing and cp gene homeology blast.The results showed that CyMV extracted from the leaves of orchid in Hangzhou,Zhejiang Province,China,could be derived from Kunming city(KM),Yunnan Province,China.This method characterized by high sensitivity,specificity,and precision is suitable for early diagnosis and quantitative detection of CyMV.
文摘Defining con-founders that affect the reliability of diagnostic tests for coronavirus disease 2019 is vital to breaking the chain of infection.The elderly population is a higher risk group for the emerging virus.However,gender seems to exert a critical role in modifying the infection risk among women owing to hormonal changes.The menopause transition is an exceptional period for older women where the protective and immunomodulatory effects of the estrogen hormone are lost.Accordingly,attention should be given to postmenopausal women since they will have an increased risk compared to their pre-menopausal peers.
文摘The B-cell lymphoma/leukemia 11A （BCL11A） gene is essential for normal lymphoid development and has been associated with hematological malignancies. In the current study, the relative expression level of BCL11A in malignant hematological cell lines was evaluated through real-time quantitative reverse transcription polymerase chain reaction （qRT-PCR）. METHODS The relative expression level of BCLllA mRNA in malignant hematological cell lines was determined through qRT- PCR using SYBR Green I dye. Glyceraldehyde-3-phosphate dehydro- genase was used as the reference gene to confirm the relative expression level of BCL11A gene mRNA. RESULTS The relative expression level of BCL11A mRNA in cell lines from B-cell malignancies was significantly higher compared with that from acute rnyeloid leukemia （P 〈 0.05）. Different cell lines with malignant B-cells exhibited a wide range of BCL11A expressions ranging from 27.37 to 93.38. CONCLUSION The overexpression of BCL11A gene mRNA in malignant B-cells might play a role in B-cell lymphoma/leukemia.
