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Objective::To investigate the effect of ancient recipe Yanghetang and its drug-contained serum on apoptosis of triple negative breast cancer cell line MDA-MB-231 based on the signal pathway of early growth response gene 1 (Egr1)/cyclin dependent inhibitor (p21). Method::The drug-containing serum of Yanghetang was prepared from rats, and MDA-MB-231 cells were cultured in vitro. The blank control group was set, and MDA-MB-231 cells in Yanghetang high, middle, and low dose groups (23.4, 11.7, 5.85 g·kg-1) were intervened for 24, 48, 72 h, respectively. After that, the cell counting kit-8(CCK-8) method was used to detect the cell proliferation of each group. The blank control group was set, while MDA-MB-231 cells in Yanghetang high, middle, and low dose groups (23.4, 11.7, 5.85 g·kg-1) were treated for 48 h, and then flow cytometry was used to detect the apoptosis of each group and the distribution of cell cycle. The expression of Egr1 and p21 mRNA in each group was detected by quantificational Real-time polymerase chain reaction (Real-time PCR), while the expression of Egr1 and p21 protein in each group was detected by Western blot. Result::After MDA-MB-231 cells were intervened by Yanghetang for 24, 48, 72 h, MDA-MB-231 cell proliferation was significantly inhibited in Yanghetang high and middle dose groups as compared with the blank control group (P<0.01). After MDA-MB-231 cells were intervened by Yanghetang for 48 h, the apoptosis was significantly increased in Yanghetang high and middle dose groups as compared with the blank control group (P<0.05, P<0.01). In the Yanghetang high, middle dose groups, the proportion of cell cycle G0/G1 phase decreased, and the proportion of G2/M phase increased (P<0.05, P<0.01). The mRNA and protein expressions of Egr1, p21 were increased in Yanghetang high and middle groups (P<0.05, P<0.01). Conclusion::Yanghetang can activate Egr1/p21 signaling pathway in MDA-MB-231 cells, increase the expression of Egr1 and p21, and cause G2/M cell cycle arrest, thereby inducing apoptosis of MDA-MB-231 cells.
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Objective: To investigate the association between early growth response gene 1 (EGR1) and Alzheimer's disease (AD) in Han Chinese people. Methods: A total of 715 AD patients and 760 health controls were recruited in two independent samples from Eastern China (382 AD patients and 426 normal individuals) and Southwest China (333 AD patients and 334 normal individuals). SNaPshot technique was utilized to analyse the single nucleotide polymorphism (SNP) of rs11743810. A public database was used to explore whether EGR1 gene was differentially expressed in the brain of AD patients and health controls. Then the protein-protein interaction (PPI) assessment was conducted using the STRING database, and the brain eQTL (expression quantitative trait loci) analysis was used to explore the difference in rs11743810 expression between different genotypes in different brain regions. Results: Cross-platform normalized data showed that there was significant difference of EGR1 expression in temporal cortex between AD patients and control subjects (|log2FC|=0.780, P=0.000 before FDR corrected; P=0.001 after FDR corrected). PPI analysis revealed that EGR1 was physically connected with amyloid precursor protein (APP) and clusterin (CLU) protein in the network. However, different genotypes of rs11743810 showed no significant difference in expression in 10 brain regions, and no significant difference in the genotype and allele frequency of rs11743810 between AD patients and controls were found in our two independent samples. Conclusion: The rs11743810 in EGR1 may not be major susceptibility gene site for AD in Han Chinese people.
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Early growth response protein (Egr) belongs to the immediate response factor , whose family members include Egr1, Egr2, Egr3 and Egr4.The Egr protein is mainly involved in the growth ,differentiation and apoptosis of cells .It is reported the Egr2/Egr3 protein can not only effect the T /B cell's proliferation and differentiation, but also participate in multiple signaling path -ways, such as JAK/STAT and NF-κB).With the in-depth study, it is found that Egr2 and Egr3 have an indispensable relationship with the occurrence of various autoimmune diseases , including Systemic Lupus Erythematosus , Scleroderma, etc.Therefore, Egr2 and Egr3 may be a potential therapy target for autoimmune diseases .This paper reviews the biological effects of Egr 2/Egr3 and its research pro-gress in relation to autoimmune diseases .
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BACKGROUND/AIMS: The role of Elk-3 in the epithelial-mesenchymal transition (EMT) during liver fibrogenesis remains unclear. Here, we determined the expression of Elk-3 in in vitro and in vivo models and in human liver fibrotic tissues. We also investigated the molecular relationships among Elk-3, early growth response-1 (Egr-1), and the mitogen activated protein kinases (MAPK) pathway during EMT in hepatocytes. METHODS: We established anin vitro EMT model in which normal mouse hepatocyte cell lines were treated with transforming growth factor (TGF)-β1 and a CCl4-induced liver fibrosis model. Characteristics of EMT were determined by evaluating the expression levels of related markers. The expression of Elk-3 and its target Egr-1 were analyzed using Western blotting. Gene silencing of Elk-3 was performed using an siRNA knockdown system. RESULTS: The expression levels of mesenchymal markers were increased during TGF-β1-induced EMT of hepatocytes. The expression levels of Elk-3 and Egr-1 were significantly (p<0.05) increased during the EMT of hepatocytes, in CCl₄-induced mouse liver fibrotic tissues, and in human liver cirrhotic tissues. Silencing of Elk-3 and inhibition of the Ras-Elk-3 pathway with an inhibitor suppressed the expression of EMT-related markers. Moreover, Elk-3 expression was regulated by p38 MAPK phosphorylation during EMT. CONCLUSIONS: Elk-3 contributes to the progression of liver fibrosis by modulating the EMT via the regulation of Egr-1 under MAPK signaling.
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Animals , Humans , Mice , Blotting, Western , Cell Line , Epithelial-Mesenchymal Transition , Gene Silencing , Hepatocytes , In Vitro Techniques , Liver Cirrhosis , Liver , Mitogen-Activated Protein Kinases , p38 Mitogen-Activated Protein Kinases , Phosphorylation , RNA, Small Interfering , Transforming Growth FactorsABSTRACT
Objective To explore the effects of early growth response gene-1 (Egr-1) on bone marrow mesenchymal stem cells (BMSC) proliferation and osteogenic differentiation,which is aimed at providing new molecular targets for the treatment of osteoporosis.Methods Bone marrow was collected from adult men and the BMSCs were cultured primarily and observed by microscope.Meanwhile,flow cytometry was used for BMSCs phenotypic identification;After transfection of pcDNA3.1/Egr-1 into BM SCs,the level of BMSCs proliferation was determined by MTT respectively on the 2 d,4 d and 6 d;On the 7 d after transfection,the ALP activity assay was used for testing the ALP activity in BMSCs.And then,alizarin red S-calcium kit was used for measuring the calcified knots respectively on the 7 d,14 d and 21 d;On the 21 d after transfection,real-time qPCR and Western blotting were used respectively for measuring the expression of mRNA and protein of Egr-1,Runx2 and NDRG1;Further,BMSCs were transfected with Egr-1 siRNA,and the content of calcium nodules,ALP activity,the expression of Egr-1,Runx2 and NDRG1 were detected as above methods.Results The cells cultured in vitro showed high level of CD90 and CD29 and very low level of CD34 and CD45,which is accorded with the characteristic of BMSCs.The pcDNA3.1/Egr-1 transfection for BMSCs had no effect on cells prolifera tion.However,the calcified knots,ALP activity and the expression of Egr 1,Runx2 and NDRG1 were increased after transfection of pcDNA3.1/Egr-1 for BMSCs.In addition,Egr-1 siRNA showed the opposite effect with pcDNA3.1/Egr-1 transfection for BMSCs.Conclusion Egr-1 induces osteogenic differentiation of BMSCs by promoting NDRG1 but has no effects on proliferation of BMSCs.
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OBJECTIVE Basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) produced by hepatocellular carcinoma (HCC) cells are responsible for the cell growth. Accumu?lating evidence shows that insulin-like growth factor-binding protein-3 (IGFBP-3) suppresses HCC cell proliferation in both IGF- dependent and independent manners. The present study is to investigate whether treatment with exogenous IGFBP-3 inhibits bFGF and PDGF production and the cell proliferation of HCC cells. METHODS Cell Counting Kit 8 assay were designed to detect HCC cell proliferation, transcription factor early growth response- 1 (EGR1) involving in IGFBP- 3 regulation of bFGF and PDGF were detected by RT-PCR and Western blot assays. Western blot assay was adopted to detect the IGFBP- 3 regulating insulin- like growth factor 1 receptor (IGF- 1R) signaling pathway. RESULTS The present study demonstrates that IGFBP-3 suppressed IGF-1-induced bFGF and PDGF expression while it does not affect their expression in the absence of IGF-1. To delineate the underlying mechanism, Western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1 (EGR1) is involved in IGFBP-3 regulation of bFGF and PDGF. IGFBP-3 inhibition of type 1 insulin-like growth factor receptor (IGF1R), ERK and AKT activation is IGF- 1- dependent. Furthermore, transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1, bFGF and PDGF expression. CONCLUSION In conclusion, these findings suggest that IGFBP- 3 suppresses transcription of EGR1 and its target genes bFGF and PDGF through inhibiting IGF- 1-dependent ERK and AKT activation. It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation, suggesting that IGFBP-3 could be a target for the treatment of HCC.
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Objective To investigate the influence of early growth response gene-1 (EGR1) on the autophagy of host cells following infection with human T cell leukemia virus type 1 (HTLV-1).MethodsA HTLV-1-positive cell line MT2 was co-cultured with HeLa cells for 24 h to construct the virus early infection model.Immunoblotting assay was used to detect the expression of HTLV-1 core protein p19 and EGR1.Luciferase reporter gene analysis was used to detect the transcriptional activity of 5′-regulatory sequence of EGR1 at different time points after co-culturing.An effective small interfering RNA (siRNA) targeting EGR1 was screened out and transfected into HeLa cells by Lipofectamine 2000.Then the transfected HeLa cells were co-cultured with the HTLV-1-positive cell line MT2 for 24 h.Immunoblotting assay was used to detect HTLV-1 core protein p19, EGR1 and autophagy-related protein LC3.Real-time PCR was performed to detect viral load.Autophagosome was analyzed by immunofluorescence after co-culturing.Results The expression of EGR1 and the transcriptional activity of pEGR1-luc gradually increased after co-culturing HeLa cells with MT2 cells for 8 h (P<0.01).The expression of EGR1 was positively correlated with host cell autophagy following HTLV-1 infection.The effective siRNA for silencing the expression of EGR1 was obtained and named as siE2.The viral load, the expression of HTLV-1 core protein p19 and the proportion of LC3B/LC3A in the co-culture model were markedly down-regulated by RNA interference with siE2, which was concomitant with a persistent decrease of intracellular autophagosome (P<0.01).Conclusion EGR1 is associated with host cell autophagy and viral replication in HTLV-1 infection.
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PURPOSE: Accumulating evidence has shown that dysregulation of microRNA-191 (miR-191) is closely associated with tumorigenesis and progression in a wide range of cancers. This study aimed to explore the potential role of miR-191 in esophageal squamous cell carcinoma (ESCC). MATERIALS AND METHODS: miR-191 expression was assessed in 93 ESCC tissue specimens by real-time polymerase chain reaction, and survival analysis was performed via Kaplan-Meier and Cox regression analyses. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, plate colony-forming, BrdU, and Transwell assays were conducted to observe the effect of miR-191 on ESCC proliferation and invasion. Luciferase reporter and western blot assays were taken to identify target genes of miR-191. RESULTS: miR-191 was overexpressed in 93 cases of ESCC, compared with adjacent normal tissues, and miR-191 expression was significantly related to differentiation, depth of invasion, TNM stage, lymph node metastasis, and distant metastasis of tumor. Kaplan-Meier and Cox regression analyses demonstrated that overexpression of miR-191 was an independent and significant predictor of ESCC prognosis. Both gain-of-function and loss-of-function experiments showed that miR-191 promoted ESCC cell proliferation and invasion activities in vitro. Early growth response 1 (EGR1), a tumor suppressor, was predicted as a direct target of miR-191. Luciferase reporter and western blot assays proved that miR-191 reduced EGR1 expression by directly binding its 3' untranslated region. Moreover, EGR1 knockdown by siRNA enhanced ESCC cell growth and invasion. CONCLUSION: Our findings provide specific biological roles of miR-191 in ESCC survival and progression. Targeting the novel miR-191/EGR1 axis represents a potential new therapeutic way to block ESCC development.
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3' Untranslated Regions , Blotting, Western , Bromodeoxyuridine , Carcinogenesis , Carcinoma, Squamous Cell , Cell Proliferation , Epithelial Cells , In Vitro Techniques , Luciferases , Lymph Nodes , Neoplasm Metastasis , Prognosis , Real-Time Polymerase Chain Reaction , RNA, Small InterferingABSTRACT
AIM:To explore whether histamine can regulate the expression of early growth response factor-1 (Egr-1) in the cerebral cortex astrocytes.METHODS:Normal wild-type (WT) mice, histidine decarboxylase knockout ( HDC-KO) mice and histamine treated HDC-KO mice were sacrificed for extracting the total RNA of the cerebral cortex. Primary cultured rat cortical astrocytes were treated with histamine at concentrations of 10 -8 , 10 -7 , 10 -6 , 10 -5 or 10 -4 mol/L for 15, 30, 60, 120 or 240 min.H1 or H2 receptor antagonists were pretreated for 15 min before histamine treat-ment.After histamine treatment, the cell total RNA or protein was extracted.The expression of Egr-1 at mRNA and protein levels was determined by real-time PCR and Western blot.RESULTS:The mRNA level of Egr-1 in cerebral cortex of HDC-KO mice was significantly lower than that in WT mice, while exogenous histamine induced the mRNA expression of Egr-1 in HDC-KO mice.In cultured astrocytes, histamine induced the mRNA expression of Egr-1.The maximum increase in the mRNA level of Egr-1 was produced by histamine at concentration of 10 -5 mol/L.In addition, histamine-induced Egr-1 mRNA accumulation peaked at 30 min after commencing stimulation, while histamine significantly increased Egr-1 protein expression at 60 min.Furthermore, histamine-induced Egr-1 expression was inhibited by H1 receptor antagonist but not H2 receptor antagonist.CONCLUSION:Histamine up-regulates the Egr-1 expression in cerebral cortex and cultured astrocytes, which may attribute to H1 receptor activation.
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Background:Hepatic veno-occlusive disease( HVOD) is a disease characterized by hepatomegaly,jaundice, ascites,weight gain and lack of effective treatment currently. Our prophase research showed that ligustrazine had therapeutic effect on Sedum aizoon induced HVOD in mice. Aims:To investigate the mechanism of therapeutic effect of ligustrazine on Sedum aizoon induced HVOD in mice. Methods:A total of 115 mice were randomly divided into 4 groups:mice in group A were intragastrically administrated with 30 mg·kg-1 ·d-1 Sedum aizoon to induce HVOD and served as model group;mice in group B were given 30 mg·kg-1 ·d-1 Sedum aizoon + 100 mg·kg-1 ·d-1 ligustrazine and served as low dose ligustrazine intervention group;mice in group C were given 30 mg·kg-1 ·d-1 Sedum aizoon + 200 mg·kg-1 ·d-1 ligustrazine and served as high dose ligustrazine intervention group;mice in group D were given 30 mg·kg-1 ·d-1 PBS and served as normal control group. After 30 days,all the mice were sacrificed. HE staining and Masson staining were performed for histological examination. The mRNA and protein expressions of tissue factor(TF),nuclear factor(NF)-κBp65 and early growth response factor( Egr)-1 in liver tissue were determined by RT-PCR and Western blotting, respectively. Results:HE staining and Masson staining histological examination showed that ligustrazine could obviously ameliorate the pathological injury of liver tissue in HVOD mice. Compared with group D,the mRNA and protein expressions of TF,NF-κBp65,Egr-1 were significantly increased in group A( P 0. 05). Conclusions:Ligustrazine has therapeutic effect on HVOD,the possible mechanism is that ligustrazine could interrupt the activation of coagulation system by reducing the expression of TF via down regulating the expressions of NF-κBp65 and Egr-1,especially in high dose ligustrazine group.
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Objective To investigate changes of serum silent information regulator 1 (Sirt1) and inflammatory cytokines in type 2 diabetes patients with different stages of diabetic nephropathy.To explore the relationship between serum Sirt1 level and inflammatory cytokines in type 2 diabetic patients with different urinary albumin excretion rates.Methods A total of 436 cases with type 2 diabetes were divided into three groups:normoalbuminuric [D1,n =168],microalbuminuric [D2;n =152],and macroalbuminuric [D3,n =116].Serum Sirt1,hypoxia-inducible factor-1α (HIF-1α),early growth response protein 1 (EGR1),insulin-like growth factors-Ⅰ (IGF-Ⅰ),and monocyte chemotactic protein-1 (MCP-1) were determined by enzyme-linked immunosorbent assay (ELISA).Results The levels of serum Sirt1 in type 2 diabetes patients were significantly lower than that in control group,and with the increase of urinary protein excretion rate,the levels of serum Sirt1 in group D1,D2 and D3 were decreased gradually (P ＜ 0.01).Compared to control,serum inflammatory cytokines (HIF-1α,EGR1,IGF-Ⅰ,and MCP-1) levels were significantly increased in type 2 diabetic patients and gradually increased in the patients of D1,D2 and D3 groups (P ＜0.01).Furthermore,Serum Sirt1 was negatively correlated with the levels of inflammatory cytokines.Age,duration,fasting blood glucose (FBG),fasting insulin (FINS),homeostasis model assessment insulin resistance index (HOMA-IR),glycosylated hemoglobin (HbA1c),low density lipoprotein (LDL),total cholesterol (TC),triglyceride (TG),serum creatinine (Scr),blood urea nitrogen (BUN),uric acid (UA),HIF-1α,EGR1,IGF-Ⅰ,and MCP-1 were positively correlated with Ln Koc of urinary albumin to creatinine ratio [Ln(ACR)] (P ＜ 0.05);and Sirt1 were negatively correlated with Ln(ACR)(P ＜ 0.01).HIF-1α,MCP-1,IGF-Ⅰ,duration,BUN,Sirt1,UA,LDL,and EGR1 were independent factors that significantly influenced Ln (ACR) (P ＜ 0.05).Conclusions Serum Sirt1 might be a new target for the treatment of diabetic nephropathy.Enhancing serum Sirt1 levels might have a role in delaying the progression of diabetic nephropathy.
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The aim of this study was to evaluate the phytotoxic activity of aqueous extracts from mature leaves, stem bark and roots of Drimys brasiliensis Miers on germination and initial seedling development of two target species, Triticum aestivum L. (wheat) and Raphanus sativus L. (radish). The aqueous extract was prepared in a proportion of 10 g of plant powder, dissolved in 100 mL of distilled water, resulting in a 10% extract concentrate. Dilutions were made with distilled water to 7.5; 5.0; 2.5% and 0% (control). Germination and seedling growth bioassays were carried out under controlled laboratory conditions. The different plant parts exerted inhibitory effects on germination and early growth of wheat and radish, however, the extract obtained from the leaves was more effective in inhibiting the germination of radish. All plant parts of D. brasiliensis may constitute a promising source in the search for compounds capable of acting as natural phytotoxins.
O objetivo deste trabalho foi avaliar o potencial fitotóxico dos extratos aquosos de folhas maduras, cascas do caule e raízes de Drimys brasiliensis Miers sobre a germinação e desenvolvimento inicial de plântulas de duas espécies receptoras, Triticum aestivumL. (trigo) e Raphanus sativus L. (rabanete). A partir do pó de cada material vegetal foram preparados extratos-tratamentos na proporção de 10 g de material vegetal para 100 mL de água destilada, produzindo-se a concentração de 10%. A partir desta, foram feitas diluições em água destilada para 7,5; 5,0; 2,5%, e 0% (controle). Bioensaios de germinação de diásporos e crescimento de plântulas foram desenvolvidos em condições controladas de laboratório. Os diferentes órgãos de D. brasiliensis exerceram efeitos inibitórios sobre os processos de germinação e crescimento inicial de trigo e rabanete, porém o extrato obtido a partir das folhas foi mais eficiente na inibição do processo de germinação do rabanete. Todos os órgãos de D. brasiliensis podem constituir uma fonte promissora na busca de compostos capazes de atuar como fitotoxinas naturais.
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Triticum , Germination , Raphanus , DrimysABSTRACT
Objective To report the clinical,electrophysiological and genetic features in a family with Charcot-Marie-Tooth disease type 1D (CMT1 D).Methods The proband,a 53-year-old man who was found with pes cavus when he was 15 years old,presented with weakness in both lower limbs at the age of 37,aggravated and numbness in legs at the age of 50.His daughter was confirmed pes cavus in her teens and weakness in both lower limbs at the age of 18.Electrophysiology and next generation sequencing were performed in the proband.Results Electrophysiological results of the proband showed demyelinating change in motor and sensory nerves.Latency prolongation was found in bilateral waves Ⅲ,V and abnormal differentiation in bilateral waves Ⅰ of brainstem auditory evoked potential,while both interpeak latencics of Ⅲ-Ⅴ were normal.DNA analysis revealed a heterozygous 1141C ＞ T mutation in exon 1 of early growth response 2 (EGR2) gene in both of the proband and his daughter.Conclusions The onset age of Arg381Cys mutation in EGR2 gene could be at juvenile with weakness in both lower limbs.The phenotype of CMT1D is mild and progressive slowly.
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AIM:To observe the effects of Egr-1 gene transfection on the expression of tumor necrosis factor-α( TNF-α) and intercellular adhesion molecule-1 ( ICAM-1) , and to investigate the role of Egr-1 in the pathogenesis of dia-betic nephropathy .METHODS:The diabetic mouse model was established .Ten mice were randomly selected as the dia-betic group .The remaining 40 mice were injected with empty plasmid , Egr-1 expression plasmid or Egr-1 siRNA plasmid via the tail vein once a week.The normal control group was also set up .The animals were sacrificed at the end of the 4th week.The renal tissues were harvested .The expressions of Egr-1, TNF-αand ICAM-1 were detected by immunohisto-chemistry and Western blot .The pathological changes were observed under electron microscope .RESULTS: In diabetic mouse kidney, the expression of Egr-1, TNF-αand ICAM-1 was increased, and irregular thickening of glomerular basement membrane , mesangial expansion and fusion of foot were observed .The change trend was more significant in Egr-1 gene transfection group , and these changes in siRNA plasmid transfection group were obviously reduced compared with diabetes group.CONCLUSION:Egr-1 up-regulates the expression of TNF-αand ICAM-1, and induces mesangial cell proliferation and mesangial extracellular matrix accumulation , which is probably one of the mechanisms of accelerating glomerulosclerosis .
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Background Flicker light can induce myopia,but its mechanism remains unclear.As one of immediate early genes,early growth response-1 (Egr-1) gene can generate rapid response to visual stimulation,however,its effect on the formation and development of myopia is below understood.Objective This study was to investigate the dynamic expression of Egr-1 gene in retinas of flicker light-induced eyes (FL) and compare the results with form deprived eyes (FD).Methods One hundred and fifty 28-day-old C57BL/6J mice were randomly assigned to the normal control group,FD group and FL group.The right eyes of mice were occluded with a semitransparent hemispherical thin plastic shell for 2 weeks in the FD group,and the right eyes of mice were stimulated by 2 Hz flicker light for 2 weeks in the FL group,and then the mice were fed in the normal light environment for 1 week.The refractive state and axial length of the model eyes were measured by murine-specific eccentric infrared photorefraction and A-scan ultrasonography before modeling and 1 hour,I day,1 week,2 weeks after modeling as well as 1 week after termination,respectively.The mice were sacrificed in above-mentioned time points to isolate the retinas.The expressions and location of Egr-1 protein and mRNA in the retinas were detected by Western blot,and reverse transcription PCR (RT-PCR) and immunochemistry.The expressions of Egr-1 markers,neuron and protein kinase C (PKC)-α,in the retinas were assayed by using immunofluorescence.The care and use of the animals followed the administration regulations for experimental animals of Jiangsu Province.Results Two weeks after modeling,the refraction of the FL group was (0.32±0.14) D,which was significantly lower than (-0.66±0.43)D in the FD group (t=6.78,P=0.00).One hour after modeling,The expression levels of Egr-1 mRNA in mouse retinas were 0.626±0.044 and 0.695±0.058 in the FD group and FL group,which were significantly declined in comparison with 1.009±0.089 of the normal group (t=14.81,P=0.01;t=9.15,P=0.03).In 2 weeks after modeling,the expression levels of Egr-1 mRNA were still lower in the FD group and F:L group compared with the normal group (all at P＜0.05).However,the expression levels were significantly elevated in the FD group and FL group compared with the normal group (t=4.13,P=0.01;t=4.26,P=0.01) at 1 week after termination.Western blot showed a dynamic decrease in the expressions of Egr-1 protein with lapse of time in the FD group and FL group with the lowest expressing level in the second week after modeling.In I week after termination of modeling,the expressing level was raised in the FD group or the FL group,but it was still lower than that ir the normal group (t =6.32,P=0.00;t =5.45,P=0.01).Egr-1 protein was mainly expressed in the retinal ganglion cell (RGC) layer,inner nuclear layer and photoreceptor layer in the normal mice,and the expression intensity was obviously weaker in the FD mice and FL mice 2 weeks after modeling.Htowever,the expression was enhanced in 1 week after termination of modeling.Neuron and PKC-α were strongly expressed in the RGCs and bipolar cells in the normal mice.Conclusions The eyes show a myopic trend after induce of flicker light in B6 mice.The expression level of Egr-1 gene in the retina down-regulates with the reduce of refraction in FL eyes,and its dynamic expressing change is consistent between the FD eyes and FL eyes.
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Early growth is associated with later health outcomes including cardiovascular disease and type 2 diabetes.It has been suggested that these associations are mediated through classical risk factors , for example dyslipidemia .It has confirmed that post-prandial levels of many risk factors have been proposed to be more important than fasting levels in the initiation and progression of a -dulthood diseases .The studies about the effect of early growth on postprandial responses have attracted considerable attention .This re-view focuses on the relationship between early growth and postprandial responses , and the role of both in adulthood diseases , aiming to further discuss the relationship and possible mechanism between early growth and later diseases .
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Objective To explore the expression of early growth response gene-1 (EGR-1) in T cells that were positive for Tax protein of human T-cell leukemia virus type 1 (HTLV-1) and its possible reg-ulatory mechanism .Methods A series of expression structures carrying the regulatory elements of EGR-1 in different length and luciferase reporter genes were constructed .TaxP cells were transfected with the con-structs containing reporter genes and cultured with 5μmol/L of NF-κB inhibitor BAY 11-7082 or equal vol-ume of DMSO.After cultured for 24 hours the cells were collected to test the luciferase activity .BAY 11-7082 or equal volume of DMSO was added into the supernatant of TaxP cell culture to test the expression of EGR-1 protein by Western blot after 24 hours of culture .Tax and its mutants M22 and M47 were transfected into 293 T cells respectively to test the expression of EGR-1 protein by Western blot after 24 hours of culture . Results The expression structures carrying the regulatory elements of EGR-1 in different length and their mutants followed by luciferase reporter genes were successfully constructed .The luciferase activity in the cells transfected with the constructs containing the elements E 1 and E2 were higher than that transfected with E3, DelE and MutE, but the reporter gene expressions were decreased with the interference of BAY 11-7082 (P<0.01).However, there were no significant changes with the luciferase activity in the cells transfected by elements E3, DelE and MutE.Western blot analysis indicated that the expression of EGR-1 protein was significantly decreased with the interference of BAY 10-7082 .The expression of EGR-1 protein in M22 mu-tants-transfected 293 T cells were decreased significantly in comparison with those by wild type tax-and M47-transfected cells .Conclusion NF-κB was the key nuclear factor in regulating the expression of EGR-1 pro-tein in Tax-positive T cells .
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Objective To construct a recombinant baculovirus dual expression vector containing NIS gene under the control of human telomerase reverse transcriptase (hTERT) promoter and plasminogen kringle 5 (K5) gene driven by early growth response 1 (Egr1) promoter,and to explore the feasibility of targeting both tumor and tumor vessel with combination of radioiodide and antiangiogenic therapy.Methods The hTERT-NIS gene and Egr1-K5 gene fragments were subcloned into baculovirus vector,then packaged and amplified in the sf9 cells to obtain recombinant baculovirus Bac-hTERT-NIS-Egr1-K5.Bac-CMV-NISEgr1-K5,Bac-hTERT-0-Egr1-K5 and Bac-hTERT-NIS-Egr1-0 were constructed as controls.The expression of NIS and K5 genes in human cervix cancers cells (HeLa) was examined by Western blot and quantitative real-time PCR.Functional NIS activity was confirmed by the uptake of 125I,the inhibition of NaClO4 and the cytotoxicity of 131I.The apoptotic effect of 131I-inducedK5 on human umbilical veins endothelial cells (HUVEC)was analyzed by an apoptosis assay using flow cytometry.Statistical analysis was performed using the analysis of variance.Results The recombinant baculovirus Bac-hTERT-NIS-Egr1-K5 was successfully constructed.The NIS gene under the control of hTERT promoter was specifically expressed in HeLa cells.The baculovirusinfected HeLa cells showed a significant increase of 125I uptake,which was significantly inhibited by NaClO4(F199.296,P＜0.05).Furthermore,a notable decreased cell survival rate (38.3％) was found after 131I treatment.The expression of K5 gene induced by 131I was elevated in a dose or time dependent manner and resulted in obvious inhibition with cell survival rate of 30.8％ in baculovirus-infected HUVEC cells,which was significantly higher than that in the control groups (11.2％ and 10.9％ respectively,F=19.926,45.409;both P＜0.05).Conclusions A recombinant baculovirus dual expression vector containing the NIS and K5 genes has been successfully constructed.This study suggests the feasibility of a synergistic strategy of NISbased raidoiodide therapy and K5-based antiangiogenic therapy in vitro,and make it possible to perform in vivo study in the near future.
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O vigor das sementes interfere significativamente na tolerância à deficiência hídrica e no rendimento de sementes. Assim, o objetivo no presente trabalho foi avaliar a influência da deficiência hídrica sobre o rendimento e a qualidade fisiológica das sementes de soja produzidas na geração F1. O trabalho foi desenvolvido utilizando-se sementes de soja da cultivar M-SOY 8008 RR, semeadas em vasos com capacidade de 15 kg de solo, coletado do horizonte A1 de um Planossolo Háplico Eutrófico solódico, em casa de vegetação. O delineamento experimental foi inteiramente casualizado, em esquema fatorial 2x2, sendo os fatores sem e com deficiência hídrica do 1° ao 10° dia após a emergência e nível de vigor das sementes (alto e baixo), totalizando quatro tratamentos, com cinco repetições. As variáveis analisadas foram a fitomassa seca da parte aérea e a área foliar aos 10, 20, 30 e 40 dias após a emergência, rendimento e qualidade fisiológica através dos testes de germinação, primeira contagem de germinação, envelhecimento acelerado e massa de mil sementes na geração F1. A utilização de sementes de alto vigor proporciona acréscimos superiores a 15% no rendimento, em relação ao uso das sementes de baixo vigor. A deficiência hídrica do 1° ao 10° dia após a emergência não interfere no rendimento e na qualidade fisiológica de sementes da geração F1, tanto daquelas de alto como de baixo vigor.
Seed vigor interferes significantly in tolerance to water deficit and seed yield. The objective of this study was to evaluate the influence of water stress on yield and physiological quality of seeds produced in the F1 generation. The study was conducted in a greenhouse using seeds of the cultivar M-SOY 8008RR, sown in pots with capacity of 15kg of soil A1 horizon of a Eutrophic Haplic Planosol Solodicin . The experimental design was completely randomized in a 2x2 factorial, with and without water deficit of 1st to 10th day after emergence and seed vigor level: high and low, totaling four treatments with five replicates. The variables analyzed were dry mass of shoots and leaf area at 10, 20, 30 and 40 days after emergence, yield and physiological quality through germination, first count, accelerated aging and weight of a thousand seeds of seeds in the F1 generation. The use of seed provides high force increases over 15% yield with respect to the use of low seed vigor. Water deficit from the 1st to the 10th day after emergence does not interfere in the yield and seed quality of the F1 generation, either for seeds of high and low vigor.
ABSTRACT
Objective To investigate the impact of hypoxia on the expression of early growth response-1 (Egr-1) and monocyte chemoattractant protein-1 (MCP-1) in mouse aorta,and to probe the underlying mechanism involving receptor for advanced glycation end-products (RAGE).Methods 3-month-old C57BL/6 mice were subjected to hypoxia [(6.0±0.5) ％ oxygen] to establish the global hypoxia model(n=6 rats for each).Aortas were dissected,Egr-1 mRNA and MCP-1 mRNA were detected by real time RT-PCR,Egr-1 and RAGE proteins were tested by Western blot,and Egr-1 DNA binding activity was assayed by electrophoretic mobility shift assay (EMSA).For blockade of RAGE,mice were pretreated with soluble RAGE (sRAGE) for 1 h by intra-peritoneal injection before they were exposed to hypoxia.Mice with normoxia were used as controls.Results After 30 minutes of hypoxic exposure,Egr-1 mRNA in aorta was increased to (28.3±0.9)folds compared with normoxic controls (F=617.17,P＜0.01),and the induction persisted for at least 3 hours.After 45 minutes of hypoxic exposure,Egr-1 proteins in aorta was increased to (5.7 ± 0.3) folds compared with normoxic controls (F =57.18,P＜ 0.01); the enhanced DNA binding activity of Egr-1 by hypoxia was attenuated by pretreatment with anti-Egr-1 lgG.After 4 hours of hypoxic exposure,MCP-1 mRNA expression in aorta was increased to(4.0±0.3)folds compared with normoxic controls (F=30.68,P＜0.01).RAGE antigen was increased significantly within 30 minutes of hypoxic exposure,with the peak at 15 minutes; hypoxia-induced Egr-1 mRNA expression was significantly attenuated by pretreatment with sRAGE (3.3 ± 0.2) folds compared with normoxic controls (F =30.20,P＜0.01).Conclusions Hypoxia significantly induces Egr-1 and MCP-1 upregulation expressions in mouse aorta,and blockade of RAGE significantly attenuates hypoxia-induced Egr-1 expression.Thcsc findings suggest RAGE signaling is involved in hypoxia-induced vascular inflammatory stress,and highlight this receptor as a potential therapeutic target to protect tissues injured by hypoxia.