In insects,20-hydroxyecdysone(20E)and insulin-like growth factor-like peptides(IGFLPs)regulate the development o f imaginal discs.However,how IGFLPs are up-regulated to impact the development of the pupal wing disc is...In insects,20-hydroxyecdysone(20E)and insulin-like growth factor-like peptides(IGFLPs)regulate the development o f imaginal discs.However,how IGFLPs are up-regulated to impact the development of the pupal wing disc is still unclear.In this study,we investigated the expression regulation of IGFLP in the pupal wing disc of silkworm,Bombyx mori.We confirmed that B.mori IGFLP(BmIGFLP)was mainly expressed in the pupal wing disc and the expression of BmIGFLP could be significantly induced by 20E.Bioinformatics analysis of BmIGFLP promoter sequence revealed three m-regulation elements(CREs)of signal transducer and activator of transcription(STAT),which is a key component in the Janus-activated kinase/STAT pathway.Luciferase activity assays showed that two CREs enhanced the transcriptional activity o f BmIGFLP.Electrophoretic mobility shift and chromatin immunoprecipitation assays demonstrated that BmSTAT proteins in the nuclear extracts o f B.mori pupal wing discs and BmN cells could only bind to the STAT CRE3,indicating that STAT CRE3 activated by BmSTAT enhances BmIGFLP expression at pupal stages.Although 20E could not enhance the expression of BmSTAT,20E enhanced the nucleus translocation o f BmSTAT to bind with the STAT CRE3 in the BmIGFLP promoter.The increase of transcriptional activity of the STAT CRE3 by overexpression of BmSTAT and addition o f 20E in BmN cells confirmed this result.Taken together,all data indicate that BmSTAT is one o f the transcription factors activating 20E-induced BmIGFLP expression in the pupal wing disc.展开更多
Actin is a ubiquitous protein and plays essential roles on cellular structure maintenance and cellular motility in both muscle and non-muscle tissues.Multiple genes encoding muscle actin have been identified from the ...Actin is a ubiquitous protein and plays essential roles on cellular structure maintenance and cellular motility in both muscle and non-muscle tissues.Multiple genes encoding muscle actin have been identified from the ascidians,including those expressed in the larval tail muscle,the adult body-wall muscle,and adult heart muscle.In this study,a novel striated non-tail muscle actin gene was identified from the RNA-seq data of Ciona savignyi embryos.Phylogenetic analysis,alignment of the N-terminal amino acid sequences and comparation of diagnostic residues provided evidence that it had high similarity with vertebrate cardiac and skeletal muscle actin.In situ hybridization and promoter-driven GFP reporter assay revealed that it was specifically expressed in the primordia of the oral and atrial siphon.We hereby defined it as siphon-specific muscle actin coding gene(Cs-SMA).A 201 bp(−1350 bp to−1150 bp)sequence containing T-box and Six1/2 binding motif within the upstream region of Cs-SMA confined the expression of GFP in the siphons of electroporated embryos.Six1/2 binding motif was experimentally confirmed to play indispensable role in controlling the siphon-specific expression of Cs-SMA.The tissue-specific expression of Cs-SMA in the siphon primordia indicated its potential crucial roles in Ciona embryogenesis and organogenesis.展开更多
Objective To investigate the combination of Epstein-Barr virus nuclear protein 3C (EBNA3C) with Gemin3 and its effect on Gemin3 expression. Methods Co-immunoprecipitation, GST pull-down and immunofluorescent assay wer...Objective To investigate the combination of Epstein-Barr virus nuclear protein 3C (EBNA3C) with Gemin3 and its effect on Gemin3 expression. Methods Co-immunoprecipitation, GST pull-down and immunofluorescent assay were used to determine the combination of EBNA3C and Gemin3 and their combining domain. Stable EBNA3C knockdown cell lines were made by lentivirus-delivered small hairpin RNA and then puromycin selection. Western blot was used to check the effect of EBNA3C on Gemin3 expression. Results EBNA3C and Gemin3 combined with each other in vivo and in vitro through their C-terminals. EBNA3C up-regulated Gemin3 gene expression. Conclusion EBNA3C forms complex with Gemin3 and up-regulates its expression.展开更多
It is observed by in situ stain that LDH (1 5) ...nNAD + can probably enter the nucleopore and can be bound bound specifically with the genes that encode them. During the in vitro expression, the dilution of heart nuc...It is observed by in situ stain that LDH (1 5) ...nNAD + can probably enter the nucleopore and can be bound bound specifically with the genes that encode them. During the in vitro expression, the dilution of heart nuclear DNA fragments could enhance the expression activity of LDH/DNA and the amount of expressed LDH (1 5) is in proportion to the amount of dissociable LDH (1 5) on the LDH/DNA. With the integration of 14C Leu to the proteins, it is also observed that the addition of LDH (1 5) ...nNAD + can suppress the in vitro expression activity of LDH/DNA. AFM observation shows that the regulation sequence at the both ends of active genes may be bound with such active factors as proteins encoded by the genes which probably is the main molecular switch of gene expression and regulation we have been always searching for. Our work shows the prospective application of the combination of AFM and isotope labeling in the research of biological reaction.展开更多
Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migra...Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migration of DCs both in vitro and in vivo. Exposure of DCs to clinical concentrations of CsA neither induces apoptosis nor alters development but does impair cytokine secretion, chemokine receptor expression, and migration. In vitro, CsA impairs the migration of mouse bone marrow-derived DCs toward macrophage inflammatory protein-3beta (MIP-3beta) and induces them to retain responsiveness to MIP-1alpha after lipopolysaccharide (LPS)-stimulated DC maturation, while in vivo administration of CsA inhibits the migration of DCs out of skin and into the secondary lymphoid organs. CsA impairs chemokine receptor and cyclooxygenase-2 (COX-2) expression normally triggered in LPS-stimulated DCs; administration of exogenous prostaglandin E2 (PGE2) reverses the effects of CsA on chemokine receptor expression and DC migration. Inhibition of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) pathway signaling by CsA may be responsible for the CsA-mediated effects on the regulation of chemokine receptor and cyclooxygenase-2 (COX-2) expression. Impairment of DC migration due to inhibition of PGE2 production and regulation of chemokine receptor expression may contribute, in part, to CsA-mediated immunosuppression.展开更多
Prostate cancer is the most commonly diagnosed cancer and the second leading cause of cancer mortality in men in the Western World. The effects of androgens are mediated by the Androgen Receptor (AR). Therefore, studi...Prostate cancer is the most commonly diagnosed cancer and the second leading cause of cancer mortality in men in the Western World. The effects of androgens are mediated by the Androgen Receptor (AR). Therefore, studies focus on the identification of AR-regulated genes that are also highly expressed in the prostate. STAMP family genes STAMP1/STEAP2 and STAMP2/STEAP4 have only expressed in androgen receptor-positive cells, the role of AR in STAMP family gene expression is an important question. STEAP (Six Transmembrane Epithelial Antigens of Prostate) is the first characterized prostate enriched six transmembrane genes, expressed in metastatic prostate cancer samples, it is tempting to speculate that STAMP/STEAP family genes may be involved in similar functions with a role for both the normal biology and pathophysiology of the prostate. Using siRNA technology in LNCaP cells expressing STAMP genes per se, an apoptosis panel including pro-apoptotic and/or apoptotic molecules was assayed by RT-PCR. In this research project, the prostate-specific STAMP gene family and its regulatory effects on the nuclear factor kappa B and caspase-related pathways were characterized. Considering that the beta-actin response in the control group was high in the immunolabeling studies, an increase in the induction of Tumor Necrosis Factor (TNF) was detected in the signals received with the vital proteins NFkB and akt, which were silenced by siRNA, which means that STAMP genes potentiate vital proteins.展开更多
MicroRNAs(miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression post-transcriptionally. Recent studies have demonstrated that miRNAs are involved in the pathogenesis of hypertrophy.We in...MicroRNAs(miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression post-transcriptionally. Recent studies have demonstrated that miRNAs are involved in the pathogenesis of hypertrophy.We investigated miR-16 expression and their potential roles in a rat model of hypertrophy induced by abdominal artery constriction (AAC).miR-16 expression was significantly decreased, and CCND1 and CCND2 protein were markedly increased without obvious change of its mRNA level after hypertrophy induction.CCND1 and CCND2 levels were increased without changing their transcript levels in neonatal rat ventricular cardiomyocytes(NRVC) induced by PE,and miR-16 was down-regulated in this process with significantly up-regulatedβ-MHC,ANF and MLC-2 expression.Conversely,introduction of functional miR-16,CCND1 siRNA or CCND2 siRNA into NRVCs could repress cardiomyocyte hypertrophy.These results implicate that miR-16 is involved in contributing to cardiac hypertrophy,one of the mechanisms may be resulted from post-transcriptional regulation of CCND1 and CCND2.展开更多
Objective: In order to explore the radioprotective effects of the expression of hematopoietic growth factors regulated by radio-inducible promoter on radiation injury. Methods:The human FL (Flt3 ligand) cDNA and EGFP ...Objective: In order to explore the radioprotective effects of the expression of hematopoietic growth factors regulated by radio-inducible promoter on radiation injury. Methods:The human FL (Flt3 ligand) cDNA and EGFP (enhanced green fluorescent protein) cDNA were linked together with IRES and then inserted into the eukaryotic expression vector pCI-Egr, which was constructed by substituting CMV promoter in pCIneo with the Egr-1 promoter (Egr-EF). The vector was transferred into human bone marrow stromal ...展开更多
Osteoarthritis (OA), the most common form of joint disease, is characterized clinically by joint pain, stiffness,and deformity. OA is now considered a whole joint disease;however, the breakdown of the articular cartil...Osteoarthritis (OA), the most common form of joint disease, is characterized clinically by joint pain, stiffness,and deformity. OA is now considered a whole joint disease;however, the breakdown of the articular cartilage remains themajor hallmark of the disease. Current treatments targeting OA symptoms have a limited impact on impeding orreversing the OA progression. Understanding the molecular and cellular mechanisms underlying OA development isa critical barrier to progress in OA therapy. Recent studies by the current authors’ group and others have revealedthat the nuclear factor of activated T cell 1 (NFAT1), a member of the NFAT family of transcription factors, regulatesthe expression of many anabolic and catabolic genes in articular chondrocytes of adult mice. Mice lacking NFAT1exhibit normal skeletal development but display OA in both appendicular and spinal facet joints as adults. Thisreview mainly focuses on the recent advances in the regulatory role of NFAT1 transcription factor in the activities ofarticular chondrocytes and its implication in the pathogenesis of OA.展开更多
BACKGROUND Sepsis is a major medical challenge.Magnolol is an active constituent of Houpu that improves tissue function and exerts strong anti-endotoxin and anti-inflammatory effects,but the mechanism by which it redu...BACKGROUND Sepsis is a major medical challenge.Magnolol is an active constituent of Houpu that improves tissue function and exerts strong anti-endotoxin and anti-inflammatory effects,but the mechanism by which it reduces intestinal inflammation in sepsis is yet unclear.AIM To assess the protective effect of magnolol on intestinal mucosal epithelial cells in sepsis and elucidate the underlying mechanisms.METHODS Enzyme-linked immunosorbent assay was used to measure tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,and regulated on activation,normal T-cell expressed and secreted(RANTES)levels in serum and ileal tissue in animal studies.The histopathological changes of the ileal mucosa in different groups were observed under a microscope.Cell Counting Kit-8 and cell permeability assays were used to determine the concentration of drug-containing serum that did not affect the activity of Caco2 cells but inhibited lipopolysaccharide(LPS)-induced decrease in permeability.Immunofluorescence and Western blot assays were used to detect the levels of RANTES,inhibitor of nuclear factor kappa-B kinaseβ(IKKβ),phosphorylated IKKβ(p-IKKβ),inhibitor of nuclear factor kappa-B kinaseα(IκBα),p65,and p-p65 proteins in different groups in vitro.RESULTS In rats treated with LPS by intravenous tail injection in the presence or absence of magnolol,magnolol inhibited the expression of proinflammatory cytokines,IL-1β,IL-6,and TNF-αin a dose-dependent manner.In addition,magnolol suppressed the production of RANTES in LPS-stimulated sepsis rats.Moreover,in vitro studies suggested that magnolol inhibited the increase of p65 nucleation,thereby markedly downregulating the production of the phosphorylated form of IKKβin LPS-treated Caco2 cells.Specifically,magnolol inhibited the translocation of the transcription factor nuclear factor-kappa B(NF-κB)from the cytosol into the nucleus and down-regulated the expression level of the chemokine RANTES in LPS-stimulated Caco2 cells.CONCLUSION Magnolol down-regulates RANTES levels by inhibiting the LPS/NF-κB signaling pathways,thereby suppressing IL-1β,IL-6,and TNF-αexpression to alleviate the mucosal barrier dysfunction in sepsis.展开更多
Upstream regulatory region and flanking DNA of yellow gene wereisolated and cloned from a Drosophila genomic library. A vector containing yellow gene and regulatory elements was constructed using the recombinant DNA t...Upstream regulatory region and flanking DNA of yellow gene wereisolated and cloned from a Drosophila genomic library. A vector containing yellow gene and regulatory elements was constructed using the recombinant DNA technique. Then this vector was integrated into Drosophila genome by genetic transformation. Using both FLP/FRT and Cre/LoxP site-specific recombination systems, two new yellow alleles were created at the same position in the genome of transgenic flies. Results from genetic and molecular analysis indicated that transcriptional enhancers regulate the developmental expression of the transgene. Furthermore, interactions between new-created yellow alleles were observed. Such interactions can influence markedly the expression of yellow gene during development. This effect may also be a form of enhancer-mediated gene expression.展开更多
Streptomyces is an important treasure trove for natural products discovery.In recent years,many scientists focused on the genetic modification and metabolic regulation of Streptomyces to obtain diverse bioactive compo...Streptomyces is an important treasure trove for natural products discovery.In recent years,many scientists focused on the genetic modification and metabolic regulation of Streptomyces to obtain diverse bioactive compounds with high yields.This review summarized the commonly used regulatory strategies for natural products discovery and overproduction in Streptomyces from three main aspects,including regulator-related strategies,promoter engineering,as well as other strategies employing transposons,signal factors,or feedback regulations.It is expected that the metabolic regulation network of Streptomyces will be elucidated more comprehensively to shed light on natural products research in the future.展开更多
The glutathione S-transferases(GSTs)are a superfamily of enzymes that function in cellular protection against toxic substances and oxidative stress.Bivalves could accumulate high concentration of paralytic shellfish t...The glutathione S-transferases(GSTs)are a superfamily of enzymes that function in cellular protection against toxic substances and oxidative stress.Bivalves could accumulate high concentration of paralytic shellfish toxins(PSTs)from harmful algae.To understand the possible involvement of GSTs in protecting bivalves during PST accumulation and metabolism,the GST genes were systemically analyzed in two cultured scallops,Azumapecten farreri and Mizuhopecten yessoensis,which were reported for PST deposition during harmful algae bloom.A total of 35 and 37 GSTs were identified in A.farreri(AfGSTs)and M.yessoensis(MyGSTs)genome,respectively,and the expansion of the sigma class from the cytosolic subfamily was observed.In both scallop species,sigma class GSTs showed higher expression than other members.The high GSTs expression was detected mainly during/after larvae stages and in the two most toxic organs,hepatopancreas and kidney.After ingesting PST-producing dinoflagellates,all the regulated AfGSTs in the hepatopancrcas were from the sigma class,but with opposite regulation pattern between Alexandrium catenella and A.minutum exposure.In scallop kidneys,where PSTs transformed into higher toxicity,more AfGSTs were regulated than in the hepatopancreas,and most of them were from the sigma class,with similar regulation pattern between A.catenella and A.minutum exposure.In M.yessoensis exposed to A.catenella,MyGST-σ2 was the only up-regulated MyGST in both hepatopancreas and kidney.Our results suggested the possible diverse function of scallop GSTs and the importance of sigma class in the defense against PSTs,which would contribute to the adaptive evolution of scallops in marine environments.展开更多
We detected loss of heterozygosity(LOH)and microsatellite instabilities(MSI),as well as extron expression of the fragile histidine triad(FHIT)gene in gastric carcinoma(GC),in order to evaluate their association with c...We detected loss of heterozygosity(LOH)and microsatellite instabilities(MSI),as well as extron expression of the fragile histidine triad(FHIT)gene in gastric carcinoma(GC),in order to evaluate their association with clinicopathological processes in gastric carcinogenesis.LOH and MSI of the FHIT were detected by using PCR at 4 microsatellite loci:D3S 1300,D3S 4103,D3S 1481,D3S 1234 in cancer tissues from 50 patients with primary GC,with normal mucosa acting as matched controls.FHIT transcripts were detected by nested RT-PCR in 30 cases of GC and their products were sequenced.Results show that the average frequencies of LOH and MSI of the FHIT gene in GC were 32.4%and 26.4%,respectively.There was no correlation between LOH and MSI of the FHIT gene in GC and the histological characteristics of gastric carcinoma(Bormann’s or Lauren’s classification).LOH of the FHIT gene in GC was related to depth invasiveness,and its frequency in GC where serosa was penetrated was significantly higher than that in GC without serosa penetration(73.5%vs 37.5%,P<0.05).The frequency of MSI in GC without lymph node metastasis was significantly higher than that in GC with lymph node metastasis(66.7%vs 34.3%,P<0.05).Aberrant transcripts were found in 11/30 GC tissues.Sequencing analysis of the aberrant fragments found a RT-PCR product missing exons 5–7 in one case of GC,and another product missing exons 4–7.Four of 10(40.0%)cases of primary GC showed absent or decreased expression of the FHIT protein as compared to their matched normal tissues.The findings in this study suggest that LOH and MSI of FHIT gene may induce aberrant extron expression,which might play a role in gastric carcinogenesis.展开更多
To investigate the expression of JWA after hemin and(or)thermal stress exposure,we treated K562(chronic myelogenous leukemia cells)cells with different doses of hemin and thermal stress using different exposure times....To investigate the expression of JWA after hemin and(or)thermal stress exposure,we treated K562(chronic myelogenous leukemia cells)cells with different doses of hemin and thermal stress using different exposure times.The expression of JWA protein was determined by Western blot analysis.Reverse transcription-polymerase chain reaction was carried out to determine JWA mRNA expression.JWA promoter transcription activity analysis was performed by chloramphenicol acetyl transferase-enzyme linked immunosorbent assay(CAT-ELISA).The expression of JWA protein was significantly increased by up to(3.23±0.57)folds compared to the control in K562 cells after hemin treatment(50μM for one week),and a similar pattern was observed in the cells after treatment with thermal stress(42℃)for 2 hours[increased by(8.00±1.73)folds].The expression of JWA mRNA was also significantly elevated by up to(1.37±0.06)folds in K562 cells treated with hemin(30μM for 48 hours),and a similar regulatory pattern[increased by(1.87±0.13)folds]was observed with thermal stress exposure(42℃)for 30 minutes.However,a combined antagonistic effect was observed in the treatment of K562 cells with hemin(30μM,48 h)followed by thermal stress(42℃,30 min).CAT-ELISA further confirmed that either hemin or thermal stress treatment could up-regulate JWA transcription activity,however,the effects could be counteracted partly by treatment with a combination of both.Hemin and thermal stress might regulate JWA expression via distinct intracellular signal transduction pathways.展开更多
Methyl-CpG binding protein 2(MeCP2)has a crucial role in transcriptional regulation and neural development(Ausio et al.,2014).Loss of function mutations of MECP2 in human lead to Rett syndrome(RTT),a severe neurodevel...Methyl-CpG binding protein 2(MeCP2)has a crucial role in transcriptional regulation and neural development(Ausio et al.,2014).Loss of function mutations of MECP2 in human lead to Rett syndrome(RTT),a severe neurodevelopmental disorders(Amir et al.,1999),whereas individuals with the chromosomal展开更多
microRNA(miRNA)is a type of small non-coding RNA that can participate in cell proliferation and apoptosis by regulating gene expression.More and more evidences indicate that miRNA-200a is involved in the occurrence an...microRNA(miRNA)is a type of small non-coding RNA that can participate in cell proliferation and apoptosis by regulating gene expression.More and more evidences indicate that miRNA-200a is involved in the occurrence and development of non-alcoholic fatty liver disease,alcoholic liver disease,drug-induced liver injury,liver fibrosis,and hepatocellular carcinoma.Downstream target genes of serotonin,regulating related signal pathways and playing different roles in the progression of a variety of liver diseases,provide a reference for exploring the mechanism of a variety of chronic liver diseases.展开更多
Pervasive transcription of the mammalian genome produces hundreds of thousands of noncoding RNAs(ncRNAs).Numerous studies have suggested that some of these ncRNAs regulate multiple cellular processes and play importan...Pervasive transcription of the mammalian genome produces hundreds of thousands of noncoding RNAs(ncRNAs).Numerous studies have suggested that some of these ncRNAs regulate multiple cellular processes and play important roles in physiological and pathological processes.Notably,a large subset of ncRNAs is enriched on chromatin and participates in regulating gene expression and the dynamics of chromatin structure and status.In this review,we summarize recent advances in the functional study of chromatin-associated ncRNAs and mechanistic insights into how these ncRNAs associate with chromatin.We also discuss the potential future challenges which still need to be overcome in this field.展开更多
SKA2(spindle and KT associated 2),also referred to as FAM33A(family with sequence similarity 33,member A),is a recently identified gene involved in cell cycle regulation,and growing evidence is implicating its roles i...SKA2(spindle and KT associated 2),also referred to as FAM33A(family with sequence similarity 33,member A),is a recently identified gene involved in cell cycle regulation,and growing evidence is implicating its roles in tumorigenesis and psychiatric disorders.It has been demonstrated that SKA2,along with its coworkers SKA1 and SKA3,constitutes the SKA complex which plays a critical role in the maintenance of the metaphase plate and/or spindle checkpoint silencing during mitosis.SKA2 is over-expressed both in cancer cell lines and clinical samples including small cell lung cancer and breast cancer,whereas downregulation of SKA2 is associated with depression and suicidal ideation.The expression of SKA2 is regulated by transcription factors including NF-kB and CREB,miRNAs as well as DNA methylation.In this review,we provide an overview of studies that reveal SKA2 gene and protein characteristics as well as physiological function,with a special focus on its transcription regulatory mechanisms,and also provide a summary regarding the translational opportunity of the SKA2 gene as a clinical biomarker for cancers and psychiatric disorders.展开更多
基金the National Basic Research Program of China(No.282012CB114603)the Scientific Research Cultivation Fund for Young Faculty of South China Normal University(Grant No.15KJ08).
文摘In insects,20-hydroxyecdysone(20E)and insulin-like growth factor-like peptides(IGFLPs)regulate the development o f imaginal discs.However,how IGFLPs are up-regulated to impact the development of the pupal wing disc is still unclear.In this study,we investigated the expression regulation of IGFLP in the pupal wing disc of silkworm,Bombyx mori.We confirmed that B.mori IGFLP(BmIGFLP)was mainly expressed in the pupal wing disc and the expression of BmIGFLP could be significantly induced by 20E.Bioinformatics analysis of BmIGFLP promoter sequence revealed three m-regulation elements(CREs)of signal transducer and activator of transcription(STAT),which is a key component in the Janus-activated kinase/STAT pathway.Luciferase activity assays showed that two CREs enhanced the transcriptional activity o f BmIGFLP.Electrophoretic mobility shift and chromatin immunoprecipitation assays demonstrated that BmSTAT proteins in the nuclear extracts o f B.mori pupal wing discs and BmN cells could only bind to the STAT CRE3,indicating that STAT CRE3 activated by BmSTAT enhances BmIGFLP expression at pupal stages.Although 20E could not enhance the expression of BmSTAT,20E enhanced the nucleus translocation o f BmSTAT to bind with the STAT CRE3 in the BmIGFLP promoter.The increase of transcriptional activity of the STAT CRE3 by overexpression of BmSTAT and addition o f 20E in BmN cells confirmed this result.Taken together,all data indicate that BmSTAT is one o f the transcription factors activating 20E-induced BmIGFLP expression in the pupal wing disc.
基金funded by the National Key Research and Development Program of China(Nos.2019YFE0190900,2018YFD0900705).
文摘Actin is a ubiquitous protein and plays essential roles on cellular structure maintenance and cellular motility in both muscle and non-muscle tissues.Multiple genes encoding muscle actin have been identified from the ascidians,including those expressed in the larval tail muscle,the adult body-wall muscle,and adult heart muscle.In this study,a novel striated non-tail muscle actin gene was identified from the RNA-seq data of Ciona savignyi embryos.Phylogenetic analysis,alignment of the N-terminal amino acid sequences and comparation of diagnostic residues provided evidence that it had high similarity with vertebrate cardiac and skeletal muscle actin.In situ hybridization and promoter-driven GFP reporter assay revealed that it was specifically expressed in the primordia of the oral and atrial siphon.We hereby defined it as siphon-specific muscle actin coding gene(Cs-SMA).A 201 bp(−1350 bp to−1150 bp)sequence containing T-box and Six1/2 binding motif within the upstream region of Cs-SMA confined the expression of GFP in the siphons of electroporated embryos.Six1/2 binding motif was experimentally confirmed to play indispensable role in controlling the siphon-specific expression of Cs-SMA.The tissue-specific expression of Cs-SMA in the siphon primordia indicated its potential crucial roles in Ciona embryogenesis and organogenesis.
文摘Objective To investigate the combination of Epstein-Barr virus nuclear protein 3C (EBNA3C) with Gemin3 and its effect on Gemin3 expression. Methods Co-immunoprecipitation, GST pull-down and immunofluorescent assay were used to determine the combination of EBNA3C and Gemin3 and their combining domain. Stable EBNA3C knockdown cell lines were made by lentivirus-delivered small hairpin RNA and then puromycin selection. Western blot was used to check the effect of EBNA3C on Gemin3 expression. Results EBNA3C and Gemin3 combined with each other in vivo and in vitro through their C-terminals. EBNA3C up-regulated Gemin3 gene expression. Conclusion EBNA3C forms complex with Gemin3 and up-regulates its expression.
文摘It is observed by in situ stain that LDH (1 5) ...nNAD + can probably enter the nucleopore and can be bound bound specifically with the genes that encode them. During the in vitro expression, the dilution of heart nuclear DNA fragments could enhance the expression activity of LDH/DNA and the amount of expressed LDH (1 5) is in proportion to the amount of dissociable LDH (1 5) on the LDH/DNA. With the integration of 14C Leu to the proteins, it is also observed that the addition of LDH (1 5) ...nNAD + can suppress the in vitro expression activity of LDH/DNA. AFM observation shows that the regulation sequence at the both ends of active genes may be bound with such active factors as proteins encoded by the genes which probably is the main molecular switch of gene expression and regulation we have been always searching for. Our work shows the prospective application of the combination of AFM and isotope labeling in the research of biological reaction.
文摘Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migration of DCs both in vitro and in vivo. Exposure of DCs to clinical concentrations of CsA neither induces apoptosis nor alters development but does impair cytokine secretion, chemokine receptor expression, and migration. In vitro, CsA impairs the migration of mouse bone marrow-derived DCs toward macrophage inflammatory protein-3beta (MIP-3beta) and induces them to retain responsiveness to MIP-1alpha after lipopolysaccharide (LPS)-stimulated DC maturation, while in vivo administration of CsA inhibits the migration of DCs out of skin and into the secondary lymphoid organs. CsA impairs chemokine receptor and cyclooxygenase-2 (COX-2) expression normally triggered in LPS-stimulated DCs; administration of exogenous prostaglandin E2 (PGE2) reverses the effects of CsA on chemokine receptor expression and DC migration. Inhibition of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) pathway signaling by CsA may be responsible for the CsA-mediated effects on the regulation of chemokine receptor and cyclooxygenase-2 (COX-2) expression. Impairment of DC migration due to inhibition of PGE2 production and regulation of chemokine receptor expression may contribute, in part, to CsA-mediated immunosuppression.
文摘Prostate cancer is the most commonly diagnosed cancer and the second leading cause of cancer mortality in men in the Western World. The effects of androgens are mediated by the Androgen Receptor (AR). Therefore, studies focus on the identification of AR-regulated genes that are also highly expressed in the prostate. STAMP family genes STAMP1/STEAP2 and STAMP2/STEAP4 have only expressed in androgen receptor-positive cells, the role of AR in STAMP family gene expression is an important question. STEAP (Six Transmembrane Epithelial Antigens of Prostate) is the first characterized prostate enriched six transmembrane genes, expressed in metastatic prostate cancer samples, it is tempting to speculate that STAMP/STEAP family genes may be involved in similar functions with a role for both the normal biology and pathophysiology of the prostate. Using siRNA technology in LNCaP cells expressing STAMP genes per se, an apoptosis panel including pro-apoptotic and/or apoptotic molecules was assayed by RT-PCR. In this research project, the prostate-specific STAMP gene family and its regulatory effects on the nuclear factor kappa B and caspase-related pathways were characterized. Considering that the beta-actin response in the control group was high in the immunolabeling studies, an increase in the induction of Tumor Necrosis Factor (TNF) was detected in the signals received with the vital proteins NFkB and akt, which were silenced by siRNA, which means that STAMP genes potentiate vital proteins.
文摘MicroRNAs(miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression post-transcriptionally. Recent studies have demonstrated that miRNAs are involved in the pathogenesis of hypertrophy.We investigated miR-16 expression and their potential roles in a rat model of hypertrophy induced by abdominal artery constriction (AAC).miR-16 expression was significantly decreased, and CCND1 and CCND2 protein were markedly increased without obvious change of its mRNA level after hypertrophy induction.CCND1 and CCND2 levels were increased without changing their transcript levels in neonatal rat ventricular cardiomyocytes(NRVC) induced by PE,and miR-16 was down-regulated in this process with significantly up-regulatedβ-MHC,ANF and MLC-2 expression.Conversely,introduction of functional miR-16,CCND1 siRNA or CCND2 siRNA into NRVCs could repress cardiomyocyte hypertrophy.These results implicate that miR-16 is involved in contributing to cardiac hypertrophy,one of the mechanisms may be resulted from post-transcriptional regulation of CCND1 and CCND2.
基金National Natural Science Foundation of China(No 39900040)Natiorlal Natural Science Outstanding Youth Foundation of China(No 39825111).
文摘Objective: In order to explore the radioprotective effects of the expression of hematopoietic growth factors regulated by radio-inducible promoter on radiation injury. Methods:The human FL (Flt3 ligand) cDNA and EGFP (enhanced green fluorescent protein) cDNA were linked together with IRES and then inserted into the eukaryotic expression vector pCI-Egr, which was constructed by substituting CMV promoter in pCIneo with the Egr-1 promoter (Egr-EF). The vector was transferred into human bone marrow stromal ...
基金supported by the United States National Institutes of Health(NIH)under Award Number R01 AR059088(to J.W.)the Mary A.and Paul R.Harrington Distinguished Professorship Endowment.
文摘Osteoarthritis (OA), the most common form of joint disease, is characterized clinically by joint pain, stiffness,and deformity. OA is now considered a whole joint disease;however, the breakdown of the articular cartilage remains themajor hallmark of the disease. Current treatments targeting OA symptoms have a limited impact on impeding orreversing the OA progression. Understanding the molecular and cellular mechanisms underlying OA development isa critical barrier to progress in OA therapy. Recent studies by the current authors’ group and others have revealedthat the nuclear factor of activated T cell 1 (NFAT1), a member of the NFAT family of transcription factors, regulatesthe expression of many anabolic and catabolic genes in articular chondrocytes of adult mice. Mice lacking NFAT1exhibit normal skeletal development but display OA in both appendicular and spinal facet joints as adults. Thisreview mainly focuses on the recent advances in the regulatory role of NFAT1 transcription factor in the activities ofarticular chondrocytes and its implication in the pathogenesis of OA.
基金Basic Public Welfare Research Foundation of Zhejiang Province,China,No.GD21H290001and Traditional Chinese Medicine Science and Technology Project Foundation of Zhejiang Province,China,No.2020ZB072.
文摘BACKGROUND Sepsis is a major medical challenge.Magnolol is an active constituent of Houpu that improves tissue function and exerts strong anti-endotoxin and anti-inflammatory effects,but the mechanism by which it reduces intestinal inflammation in sepsis is yet unclear.AIM To assess the protective effect of magnolol on intestinal mucosal epithelial cells in sepsis and elucidate the underlying mechanisms.METHODS Enzyme-linked immunosorbent assay was used to measure tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,and regulated on activation,normal T-cell expressed and secreted(RANTES)levels in serum and ileal tissue in animal studies.The histopathological changes of the ileal mucosa in different groups were observed under a microscope.Cell Counting Kit-8 and cell permeability assays were used to determine the concentration of drug-containing serum that did not affect the activity of Caco2 cells but inhibited lipopolysaccharide(LPS)-induced decrease in permeability.Immunofluorescence and Western blot assays were used to detect the levels of RANTES,inhibitor of nuclear factor kappa-B kinaseβ(IKKβ),phosphorylated IKKβ(p-IKKβ),inhibitor of nuclear factor kappa-B kinaseα(IκBα),p65,and p-p65 proteins in different groups in vitro.RESULTS In rats treated with LPS by intravenous tail injection in the presence or absence of magnolol,magnolol inhibited the expression of proinflammatory cytokines,IL-1β,IL-6,and TNF-αin a dose-dependent manner.In addition,magnolol suppressed the production of RANTES in LPS-stimulated sepsis rats.Moreover,in vitro studies suggested that magnolol inhibited the increase of p65 nucleation,thereby markedly downregulating the production of the phosphorylated form of IKKβin LPS-treated Caco2 cells.Specifically,magnolol inhibited the translocation of the transcription factor nuclear factor-kappa B(NF-κB)from the cytosol into the nucleus and down-regulated the expression level of the chemokine RANTES in LPS-stimulated Caco2 cells.CONCLUSION Magnolol down-regulates RANTES levels by inhibiting the LPS/NF-κB signaling pathways,thereby suppressing IL-1β,IL-6,and TNF-αexpression to alleviate the mucosal barrier dysfunction in sepsis.
基金the National High Technology Program For Young Scientists and Life Science Specific Fund (stz98-2-05) and Knowledge Innovation Foundation of Chinese Academy of Sciences.
文摘Upstream regulatory region and flanking DNA of yellow gene wereisolated and cloned from a Drosophila genomic library. A vector containing yellow gene and regulatory elements was constructed using the recombinant DNA technique. Then this vector was integrated into Drosophila genome by genetic transformation. Using both FLP/FRT and Cre/LoxP site-specific recombination systems, two new yellow alleles were created at the same position in the genome of transgenic flies. Results from genetic and molecular analysis indicated that transcriptional enhancers regulate the developmental expression of the transgene. Furthermore, interactions between new-created yellow alleles were observed. Such interactions can influence markedly the expression of yellow gene during development. This effect may also be a form of enhancer-mediated gene expression.
基金This work was supported by the National Key R&D Program of China(2018YFA0903300)the Natural Science Foundation of Tianjin Province(19JCYBJC24200)the National Natural Science Foundation of China(81502966).
文摘Streptomyces is an important treasure trove for natural products discovery.In recent years,many scientists focused on the genetic modification and metabolic regulation of Streptomyces to obtain diverse bioactive compounds with high yields.This review summarized the commonly used regulatory strategies for natural products discovery and overproduction in Streptomyces from three main aspects,including regulator-related strategies,promoter engineering,as well as other strategies employing transposons,signal factors,or feedback regulations.It is expected that the metabolic regulation network of Streptomyces will be elucidated more comprehensively to shed light on natural products research in the future.
基金This work was funded by the National Key R&D Program of China(2019YFC1605704)the National Natural Science Foundation of China(31630081 and 31802292)Taishan Industry Leading Talent Project.
文摘The glutathione S-transferases(GSTs)are a superfamily of enzymes that function in cellular protection against toxic substances and oxidative stress.Bivalves could accumulate high concentration of paralytic shellfish toxins(PSTs)from harmful algae.To understand the possible involvement of GSTs in protecting bivalves during PST accumulation and metabolism,the GST genes were systemically analyzed in two cultured scallops,Azumapecten farreri and Mizuhopecten yessoensis,which were reported for PST deposition during harmful algae bloom.A total of 35 and 37 GSTs were identified in A.farreri(AfGSTs)and M.yessoensis(MyGSTs)genome,respectively,and the expansion of the sigma class from the cytosolic subfamily was observed.In both scallop species,sigma class GSTs showed higher expression than other members.The high GSTs expression was detected mainly during/after larvae stages and in the two most toxic organs,hepatopancreas and kidney.After ingesting PST-producing dinoflagellates,all the regulated AfGSTs in the hepatopancrcas were from the sigma class,but with opposite regulation pattern between Alexandrium catenella and A.minutum exposure.In scallop kidneys,where PSTs transformed into higher toxicity,more AfGSTs were regulated than in the hepatopancreas,and most of them were from the sigma class,with similar regulation pattern between A.catenella and A.minutum exposure.In M.yessoensis exposed to A.catenella,MyGST-σ2 was the only up-regulated MyGST in both hepatopancreas and kidney.Our results suggested the possible diverse function of scallop GSTs and the importance of sigma class in the defense against PSTs,which would contribute to the adaptive evolution of scallops in marine environments.
基金This project was supported by the National Natural Sciences Foundation of China(Grant Nos.30600286,No.30371607).
文摘We detected loss of heterozygosity(LOH)and microsatellite instabilities(MSI),as well as extron expression of the fragile histidine triad(FHIT)gene in gastric carcinoma(GC),in order to evaluate their association with clinicopathological processes in gastric carcinogenesis.LOH and MSI of the FHIT were detected by using PCR at 4 microsatellite loci:D3S 1300,D3S 4103,D3S 1481,D3S 1234 in cancer tissues from 50 patients with primary GC,with normal mucosa acting as matched controls.FHIT transcripts were detected by nested RT-PCR in 30 cases of GC and their products were sequenced.Results show that the average frequencies of LOH and MSI of the FHIT gene in GC were 32.4%and 26.4%,respectively.There was no correlation between LOH and MSI of the FHIT gene in GC and the histological characteristics of gastric carcinoma(Bormann’s or Lauren’s classification).LOH of the FHIT gene in GC was related to depth invasiveness,and its frequency in GC where serosa was penetrated was significantly higher than that in GC without serosa penetration(73.5%vs 37.5%,P<0.05).The frequency of MSI in GC without lymph node metastasis was significantly higher than that in GC with lymph node metastasis(66.7%vs 34.3%,P<0.05).Aberrant transcripts were found in 11/30 GC tissues.Sequencing analysis of the aberrant fragments found a RT-PCR product missing exons 5–7 in one case of GC,and another product missing exons 4–7.Four of 10(40.0%)cases of primary GC showed absent or decreased expression of the FHIT protein as compared to their matched normal tissues.The findings in this study suggest that LOH and MSI of FHIT gene may induce aberrant extron expression,which might play a role in gastric carcinogenesis.
基金This work was supported partially by the National Natural Science Foundation of China(No.30471430)National Key Program(973)for Basic Research(No.2002CB512900).
文摘To investigate the expression of JWA after hemin and(or)thermal stress exposure,we treated K562(chronic myelogenous leukemia cells)cells with different doses of hemin and thermal stress using different exposure times.The expression of JWA protein was determined by Western blot analysis.Reverse transcription-polymerase chain reaction was carried out to determine JWA mRNA expression.JWA promoter transcription activity analysis was performed by chloramphenicol acetyl transferase-enzyme linked immunosorbent assay(CAT-ELISA).The expression of JWA protein was significantly increased by up to(3.23±0.57)folds compared to the control in K562 cells after hemin treatment(50μM for one week),and a similar pattern was observed in the cells after treatment with thermal stress(42℃)for 2 hours[increased by(8.00±1.73)folds].The expression of JWA mRNA was also significantly elevated by up to(1.37±0.06)folds in K562 cells treated with hemin(30μM for 48 hours),and a similar regulatory pattern[increased by(1.87±0.13)folds]was observed with thermal stress exposure(42℃)for 30 minutes.However,a combined antagonistic effect was observed in the treatment of K562 cells with hemin(30μM,48 h)followed by thermal stress(42℃,30 min).CAT-ELISA further confirmed that either hemin or thermal stress treatment could up-regulate JWA transcription activity,however,the effects could be counteracted partly by treatment with a combination of both.Hemin and thermal stress might regulate JWA expression via distinct intracellular signal transduction pathways.
基金supported by the Strategic Priority Research Program of Chinese Academy of Sciences(Grant No.XDB02050400)the National Natural Science Foundation of China(Grant No. #91432111) to Z.Qiu
文摘Methyl-CpG binding protein 2(MeCP2)has a crucial role in transcriptional regulation and neural development(Ausio et al.,2014).Loss of function mutations of MECP2 in human lead to Rett syndrome(RTT),a severe neurodevelopmental disorders(Amir et al.,1999),whereas individuals with the chromosomal
基金National Natural Science Foundation of China(No.81860790)Guangxi Science and Technology Project(No.Guike AB20297002)+3 种基金Guangxi Natural Science Foundation(No.2020GXNSFAA297160)Guangxi Natural Science Foundation(No.2018GXNSFBA050050)Guangxi First-class Discipline Integrated Traditional Chinese and Western Medicine Cultivation Discipline(No.2019XK159)Guangxi Special Expert Special Project Funding(No.Gui Ren Cai Tong Zi(2019)13)。
文摘microRNA(miRNA)is a type of small non-coding RNA that can participate in cell proliferation and apoptosis by regulating gene expression.More and more evidences indicate that miRNA-200a is involved in the occurrence and development of non-alcoholic fatty liver disease,alcoholic liver disease,drug-induced liver injury,liver fibrosis,and hepatocellular carcinoma.Downstream target genes of serotonin,regulating related signal pathways and playing different roles in the progression of a variety of liver diseases,provide a reference for exploring the mechanism of a variety of chronic liver diseases.
基金the National Natural Science Foundation of China(31925015,32122019,32270582).
文摘Pervasive transcription of the mammalian genome produces hundreds of thousands of noncoding RNAs(ncRNAs).Numerous studies have suggested that some of these ncRNAs regulate multiple cellular processes and play important roles in physiological and pathological processes.Notably,a large subset of ncRNAs is enriched on chromatin and participates in regulating gene expression and the dynamics of chromatin structure and status.In this review,we summarize recent advances in the functional study of chromatin-associated ncRNAs and mechanistic insights into how these ncRNAs associate with chromatin.We also discuss the potential future challenges which still need to be overcome in this field.
基金This work was supported by the National Natural Science Foundation of China(No.81672301 to Youquan Bu)the Basic Sciences and Advanced Technology Key Project of CQ CSTC(No.cstc2017jcyjBX0069 to Youquan Bu).
文摘SKA2(spindle and KT associated 2),also referred to as FAM33A(family with sequence similarity 33,member A),is a recently identified gene involved in cell cycle regulation,and growing evidence is implicating its roles in tumorigenesis and psychiatric disorders.It has been demonstrated that SKA2,along with its coworkers SKA1 and SKA3,constitutes the SKA complex which plays a critical role in the maintenance of the metaphase plate and/or spindle checkpoint silencing during mitosis.SKA2 is over-expressed both in cancer cell lines and clinical samples including small cell lung cancer and breast cancer,whereas downregulation of SKA2 is associated with depression and suicidal ideation.The expression of SKA2 is regulated by transcription factors including NF-kB and CREB,miRNAs as well as DNA methylation.In this review,we provide an overview of studies that reveal SKA2 gene and protein characteristics as well as physiological function,with a special focus on its transcription regulatory mechanisms,and also provide a summary regarding the translational opportunity of the SKA2 gene as a clinical biomarker for cancers and psychiatric disorders.
