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Aim To investigate the regulation and mechanism of inflammatory cytokines interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) on the drug metabolism enzyme cytochrome P450 2E1 (CYP2E1) during immune-mediated liver injury. Methods By injection of Bacillus Calmette-Guerin(BCG, 125 mg·kg-1) prepared rat model of immunological liver damage. To detection of CYP2E1 probe drug chlorzoxazone in blood concentration changes of rats over time reflects its metabolic activity by high-performance liquid chromatography (HPLC) assay. The changes of IL-1β and TNF-α in liver tissue of rats were detected by ELISA. The expression of CYP2E1 protein was determined by Western blot analysis. Results After 14 days of BCG injection of rats, a large number of mononuclear cells and lymphocytes infiltrated around the liver parenchyma and the portal area, resulting in a large number of granuloma masses with different sizes and diffuse distribution, IL-1β and TNF-α high expression (P < 0.01), CYP2E1 metabolic activity and protein expression were significantly decreased (P < 0.05). The protein content of CYP2E1 was significantly negatively correlated with IL-1β (r=0.222, P=0.027). Activation of PDTC-mediated nuclear factor-kappa B (NF-κB) inhibited the high expression of IL-1β and TNF-α in liver tissue and slowed down the down-regulation of CYP2E1 metabolism and protein expression. Conclusions NF-κB may be involved in the regulation of CYP2E1 by regulating the inflammatory cytokines IL-1β and TNF-α.
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Aim To investigate the anti-inflammatory effect of salidroside on lipopolysaccharide (LPS)-induced BV2 microglia and its mechanism. Methods The model of LPS-induced injury in BV2 microglia was established. The expression of cytokines IL-6, IL-1β and TNF-α mRNA was detected by qPCR, and the expression of Akt, p-Akt, nuclear protein NF-κB p50 protein was dectected by Western blot with different concentrations of salidroside treatment. The model was sequentially treated with PI3K inhibitor LY294002 for 30 min, and after the treatment of salidroside, the expressions of Akt, p-Akt, nuclear protein NF-κB p50, IL-6, IL-1β, TNF-α and other indicators were dectected by Western blot. Results Compared with model, salidroside reduced the expression of IL-6, IL-1β and TNF-α mRNA in BV2 microglia, improved the expression of p-Akt protein, and reduced the nuclear protein NF-κB p50. After the treatment of LY294002, salidroside had no obvious effect on the expression of p-Akt, nuclear protein NF-κB p50, IL-6, IL-1β. Conclusions Salidroside can inhibit LPS-induced BV2 microglia inflammatory reaction by activating PI3K/Akt signaling pathway, promoting Akt phosphorylation, inhibiting NF-κB p50 nuclear transcription, and inhibiting cytokines.
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Background & objectives: Scrub typhus is a chigger-borne disease caused by Orientia tsutsugamushi. The immunological reactions to O. tsutsugamushi infection are not completely understood. In this study, we investigated the response of dendritic cells (DCs) to a major 56-kDa scrub typhus antigen Sta56. Methods: Monocyte-derived human DCs were incubated with different concentrations of recombinant Sta56 and analyzed for maturation based on phagocytic capacity, the ability to induce T-cell proliferation, expression of surface markers, cytokine secretion and activation of toll-like receptor (TLR)-dependent signalling pathways. Results: Treatment of DCs with Sta56 induced cell surface expression of CD80, CD83, CD86 and MHC Class II increased the production of interleukin-12 (IL-12) p40, IL-12 p70 and IL-10 and decreased DC phagocytic capacity. Furthermore, Sta56 increased the ability of DCs to activate T-cell proliferation and interferon-? secretion. TLR4-specific antibodies neutralized Sta56-elicited effects on DC maturation, suggesting direct interaction between Sta56 and TLR4. Moreover, Sta56 activated nuclear factor (NF)-?B and p38 mitogen-activated protein kinase (MAPK) signalling as evidenced by decrease in Sta56-induced cytokine production and surface marker expression by specific inhibitors helenalin and SB203580, respectively, and increase in I?B? and p38 phosphorylation and NF-?B-DNA binding. Interpretation & conclusions: Our results showed that the surface antigen of O. tsutsugamushi activated DCs through interaction with TLR4 and activation of MAPK and NF-?B signalling, suggesting Sta56 as a potential candidate molecule for the development of vaccine against scrub typhus.
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Background: Diabetic nephropathy is a leading cause of end-stage renal failure worldwide. Purpose: The methanol leaf extract of Vernonia amygdalina (MLVA) was thus investigated for its nephroprotective effects in diabetes.Materials and Methods: Diabetes was induced in male Wistar rats by a single intraperitoneal (I.P) injection of a freshly prepared solution of Alloxan monohydrate (100 mg/kg). Forty-eight hours after alloxan administration, rats with fasting blood glucose levels of 200 mg/dl and above were used for the study. Animals were grouped into five (A-E) of nine animals each. Group A was non-diabetic non treated control; Group B animals were the diabetic untreated control rats while groups C, D and E animals were diabetic and treated with glibenclamide, MLVA 200 mg/kg and MVLA 400 mg/kg respectively. Biochemical changes were evaluated by measuring the serum markers of kidney damage (creatinine and blood urea nitrogen). Markers of oxidative stress and antioxidant activities were measured in renal tissues. Histopathological and immunohistochemical changes were also evaluated.Results: Four-week administration of MLVA produced significant (p<0.05) decrease in serum creatinine, urea, and oxidative markers but it caused a significant increase in enzymic and nonenzymic antioxidant as well as downregulation of Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-?B) and upregulation of B-cell lymphoma 2 (Bcl-2).Conclusion: MLVA ameliorates diabetic nephropathy through its antioxidant, anti-inflammatory and antiapoptotic effects
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Cocaine is a widely used drug and its abuse is associated with physical, psychiatric and social problems. Abnormalities in newborns have been demonstrated to be due to the toxic effects of cocaine during fetal development. The mechanism by which cocaine causes neurological damage is complex and involves interactions of the drug with several neurotransmitter systems, such as the increase of extracellular levels of dopamine and free radicals, and modulation of transcription factors. The aim of this review was to evaluate the importance of the dopaminergic system and the participation of inflammatory signaling in cocaine neurotoxicity. Our study showed that cocaine activates the transcription factors NF-κB and CREB, which regulate genes involved in cellular death. GBR 12909 (an inhibitor of dopamine reuptake), lidocaine (a local anesthetic), and dopamine did not activate NF-κB in the same way as cocaine. However, the attenuation of NF-κB activity after the pretreatment of the cells with SCH 23390, a D1 receptor antagonist, suggests that the activation of NF-κB by cocaine is, at least partially, due to activation of D1 receptors. NF-κB seems to have a protective role in these cells because its inhibition increased cellular death caused by cocaine. The increase in BDNF (brain-derived neurotrophic factor) mRNA can also be related to the protective role of both CREB and NF-κB transcription factors. An understanding of the mechanisms by which cocaine induces cell death in the brain will contribute to the development of new therapies for drug abusers, which can help to slow down the progress of degenerative processes.
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A sepse é a principal causa de morte em unidades de terapia intensiva (UTIs) no mundo. A reduzida disponibilidade do aminoácido mais abundante do organismo, a glutamina contribui para o complicado estado catabólico da sepse. No presente estudo investigamos os efeitos da suplementação oral com L-glutamina e L-alanina (GLN+ALA), ambos na norma livre e como dipeptídeo, L-alanil-L-glutamina (DIP), sobre o eixo glutamina-glutationa (GSH), sistema imune, inflamação, proteínas de choque térmico (HSPs) e expressão de genes envolvidos com vias de sinalização proteica em animais endotoxêmicos. Camundongos C57/B6 foram submetidos à endotoxemia (Escherichia coli LPS, 5 mg.kg-1, grupo LPS) e suplementados por 48 horas com L-glutamina (1 g.kg-1) e L-alanina (0,61 g.kg-1, grupo GLN+ALA-LPS) ou 1,49 g.kg-1 de DIP (grupo DIP-LPS). A endotoxemia promoveu depleção da concentração de glutamina no plasma (71%), músculo esquelético (50%) e fígado (49%), quando comparado ao grupo CTRL, sendo restauradas nos grupos DIP-LPS e GLN+ALA-LPS (P<0,05), fato que atenuou a redução da GSH e o estado redox (taxa GSSG/GSH) em eritrócitos circulantes, musculo e fígado (P<0,05). A suplementação em animais endotoxêmicos resultou em uma upregulation dos genes GSR, GPX1 e GCLC no músculo e fígado. A concentração das citocinas plasmáticasTNF-α, IL-6, IL-1ß e IL-10 foi atenuada pelas suplementações, bem como a expressão de mRNAs envolvidos com a resposta inflamatória, ativadas pela via do NF-κB(P<0,05). Concomitantemente, verificou-se aumento da capacidade proliferativa de linfócitos T e B circulantes nos grupos GLN+ALA-LPS e DIP-LPS. A expressão de mRNAs e a concentração de HSPs no tecido muscular foi restabelecida pelas suplementações, contudo, a expressão mRNAs relacionados às vias de síntese e degradação proteica foi somente estimulada no tecido hepático(P<0,05). Os resultados do presente estudo demonstram que a suplementação por via oral com GLN+ALA ou DIP podem ser utilizados clinicamente como métodos nutricionais em reverter o quadro de depressão da disponibilidade de glutamina corporal da sepse induzida por LPS, tendo impacto no eixo glutamina-glutationa, sistema imune e inflamatório
Sepsis is the leading cause of death inintensive care units (ICUs) in the world.The availability ofthe most abundant amino acid in the body, glutamine, is reduced in this situation, fact that contribute to the complicated catabolic state of sepsis. In the present study, we investigated the effects of oral supplementation with L-glutamine and L-alanine (GLN+ALA), both in their free form and as a dipeptide, L-alanyl-L-glutamine (DIP) on glutamine-glutathione axis (GSH), immune and inflammatory system, heat shock proteins (HSPs) expression and gene expressions involved in protein signaling pathways during endotoxemia. C57/B6 mice were subjected to endotoxemia (Escherichia coli LPS, 5 mg.kg-1, LPS group) and supplemented for 48 hours with L-glutamine (1 g.kg-1) plus L-alanine(0.61 g.kg-1, GLN+ALA-LPS group) or 1.49 g.kg-1of DIP (DIP-LPS group). Endotoxemia promoted depletion glutamine concentration in plasma (71%), skeletal muscle (50%) and liver (49%), when compared to the CTRL group, and was restored in the DIP-LPS e GLN+ALA-LPS (P<0.05), fact that attenuate the reduction of GSH and the redox state (GSSG/GSH rate) in circulating erythrocytes, liver and muscle (P<0.05). Supplementations in endotoxemic mice resulted in upregulation of GSR, GCLC and GPX1 genes in muscle and liver. Plasma concentration of TNF-α, IL-6, IL-1ß and IL-10 were attenuated by supplementation as well as the expression of mRNAs involved in the inflammatory response, activated by NFκ-B pathway (P <0.05). At the same time, high proliferative capacity of circulating T and B lymphocytes GLN+ALA-LPS e DIP-LPS were observed. HSPs (protein and mRNAs) and in muscle were restored by the supplements, however, the mRNAs expression related to the synthesis and degradation of protein pathways was only stimulated in the liver (P <0.05). Our results demonstrate that oral supplementation with GLN+ALA or DIP can be used as clinically nutritional methods to reverse the depression of body glutamine availability during sepsis induced by LPS, impacting on the glutamine-glutathione axis, immune and inflammatory system
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Animals , Mice , Endotoxemia/blood , Dipeptides/adverse effects , Glutamine/adverse effects , Immune System/abnormalities , Amino Acids , Glutathione Transferase , Heat-Shock Proteins , Nutritional and Metabolic DiseasesABSTRACT
Objective: To investigate the effects of transfection of antisense oligodeoxynucleotide (ASODN) of nuclear transcription factor κB (NF-κB) on tumorigenesis and angiogenesis of human gastric cancer cell line SGC7901 in nude mice. Methods: ASODN targeted P65 promoter region of NF-κB was designed, synthesized, and transfected into gastric cancer cell line SGC-7901. Fluorescent staining was used to observe transfection efficacy. The growth-inhibiting rate of cells was measured by MTT assay in vitro. The subcutaneous xenograft model of human gastric cancer cells was established in nude mice and the tumor growth curve was observed. The microvessel density (MVD) was detected by HE staining, and the apoptotic index (AI) was quantitatively determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL). Results: The cell proliferation was significantly inhibited in P65 ASODN-transfected group in vitro (P < 0.05). In vivo tumor formation test showed that the tumor volume in nude mice in ASODN group was obviously smaller than that of other groups (P < 0.05) and the AI was significantly higher than that of other groups (P < 0.001). The MVD in ASODN group was markedly lower than that of other groups (P < 0.01). Conclusion: Transfection of P65 ASODN significantly inhibited the proliferation of SGC-7901 cells in vitro. The mechanism may be related to induction of cell apoptosis and inhibition of angiogenesis caused by P65 ASODN.
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Progesterone is an essential hormone for pregnancy maintenance. This hormone acts by binding to its intracellular receptor or by rapid non-genomic actions to regulate a wide variety of biological functions in the feto-placental unit. Progesterone regulates blastocyst implantation and placental development by inducing immunosupression through type Th2 cytokines secretion. This review summarizes current research about the role of progesterone as critical regulator of expression and secretion of cytokines by T-cell and other placental cells.
La progesterona es una hormona esteroide muy versátil y esencial para el mantenimiento del embarazo. El principal mecanismo de acción de la progesterona es el clásico, vía receptor intracelular, regulando diversas funciones, aspectos celulares y vías moleculares implicadas en el proceso de la implantación. Asimismo existen mecanismos adicionales que no dependen de la interacción del complejo hormona receptor con la maquinaria transcripcional y que son capaces de regular rápidamente cascadas de señalización que determinarán la respuesta de la célula. En particular se ha demostrado que la progesterona ejerce efectos inmunosupresores durante la gestación al favorecer la secreción de citocinas de tipo Th2 por los linfocitos T, evento importante para regular el sistema inmunológico materno y evitar el rechazo de la placenta. El objetivo de esta revisión se centra en analizar la influencia de la progesterona en la interfase materno-fetal sobre la expresión y secreción de citocinas por las células T y no T como es el caso del trofoblasto.
Subject(s)
Animals , Female , Mice , Pregnancy , Pregnancy Maintenance/immunology , Progesterone/physiology , Blastocyst , Cytokines/physiology , Embryo Implantation/immunology , Embryo Implantation/physiology , Gene Expression Regulation , Immune Tolerance , Inflammation , Labor, Obstetric/physiology , Lymphocytes/metabolism , Models, Biological , Maternal-Fetal Exchange/immunology , NF-kappa B/physiology , Placenta/growth & development , Placenta/immunology , Pregnancy Maintenance/physiology , Pregnancy Proteins/physiology , Receptors, Progesterone/physiology , Suppressor Factors, Immunologic , Spleen/metabolismABSTRACT
Objective To study the adverse effects of PM2.5 from Beijing urban areas on human lung adenocarcinoma cells(A549) and the expression of NF-?B in the cells. Methods A549 cells were treated with PM2.5 at 25,50,100 and 200 ?g/ml for 24 h. Cytotoxicity of PM2.5 was measured by MTT assay. The activity of NF-?B was measured by ELISA assay. Western blot method was used to detect the expression of NF-?B and NO levels was determined by using the nitrate reductase method. Results PM2.5 could induce A549 cell proliferation at low doses,but inhibit cell proliferation at high doses. The activity of NF-?B increased in the cell nucleus,but decreased in the cytoplasm after exposure to PM2.5 with a significant dose-dependent manner(P
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OBJECTIVE:To explore the effect of sodium selenite intervention on the activity of NF-?B in renal tissue of rats with different stages of diabetes mellitus(DM).METHODS:SD rats were randomly divided into these groups after the diabetics model were established successfully:DM2 group(DM rats at 2 months),DM4 group(DM rats at 4 months),DM+Se2 group(Se-treated DM rats at 2 months),DM+Se4 group(Se-treated DM rats at 4 months),and set up CON2 group(normal control group at 2 months)and CON4 group(normal control group at 4 months)stimultaneously.All these groups were treated by corresponding methods after feeding different diets for 2 or 4 months,UAER was computed;Scr,BUN,Ucr and the activity of NF-?B in renal tissue were detected.RESULTS:UAER level and activity of NF-?B in DM+Se2 group were all higher than in CON2 group(P
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Objective:To investigate the expression of NF?B p65 and PTEN in laryngeal squamous cell carcinoma(LSCC) and their clinical significance.Methods:Immunohistochemical staining was used to examine the expression of NF?B p65 and PTEN in 52 cases of LSCC and 30 cases of adjacent normal tissues.Results:The expression positive rates of NF?B p65 in LSCC were obviously higher than those in adjacent normal tissues(P0.05).The Spearman analysis indicated that the expression level of NF?B p65 was negatively correlated with that of PTEN significantly(rs=-0.580,P
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16.7mmol/L)48hours after injection.The successful diabetic rats models were randomly divided into diabetic mellitus(DM) group and taurine(TA) group.DM and TA groups were subdivided into DM1,DM2 and TA1,TA2 groups according to the duration of diabetes.The expression of Caspase-3 and NF-?B mRNA were studied by reverse-transcriptase polymerase chain reaction(RT-PCR) method.Results:NO obvious expression of Caspase-3 and NF-?B mRNA were found in the nomal groups compared with DM group after the 4th and 8th week.Under the intervention of taurine,the expression of Caspase-3 and NF-?B mRNA were inhibited compared with those in model.group(P
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NF-?B, a group of important transcription factors, are introduced and discussed here on several aspects: their component and molecular structure; their activity control by I?B and IKK, the mechanism of their activation; their important roles in transcriptional regulation for large numbers of genes; and their importance in immunity, inflammation, cell survival, proliferation and apoptosis. The analysis of the relation between NF-?B and disease occurrence, the analysis of the relation between NF-?B and disease therapy, and the application prospect of the new strategy regarding the novel drug design and correlative diseases therapy on the basis of NF-?B as the target, are also included.
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Mice was stimulated by peptidoglycan from Lactobacillus sp and detect cytokines production。It was found that PG induced the production of inflammatory cytokines (IL-1,TNF-? in peritoneal macrophages,IFN-? in spleen cell)and did not induce the IL-2 production in spleen cell. Affymetrix MOE430A genechip was used to analyze changed gene expression of immune cells. It was found that expression of cytokines and related genes were changed under peptidoglycan administration. This might induced by activation of TLR-NF-?B signal pathway.
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Objective To probe the effects of nuclear factor kappa B (NF-?B) on cell apoptosis and left ventricular segmental function in acute ischemia repeffusion in dogs.Method Twenty-four dogs were randomly divided into three groups:without left anterior artery (lAD) ligation group (C group),LAD was occluded 30 min following reperfusion 120 minutes in isehemical reperfusion group (IR group),and dogs were administered with PDTC before LAD ligation in ischemical reperfusion plus pyorrole dithitocarbamate group (PDTC group).The left ventricular segmental function was detected by echo cardiography using strain rate anlysis software.EF measured by Simpson's method.Cardiac myocyte apoptosis numbers were determined by terminal deoxynudeotidy transferease-mediated biotinylated deoxyuridine triphosphate nick end labeling (TUNEL).lmmunohistochemistry and western-blot anylysis of NF-?B protein expression.Results NF-?B was obviously expression on injury myocardium of IR group,and increased significantly in contrast to control group (P0.05)Conclusions NF-?B might play an important role in acute myocardial ischemia reperfusion.PDTC reduces myocardial iscbemia/repeffasion injury by preventing expression of factor NF-?B.
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Objective To investigate the effects of Ginaton on acute lung injury (ALI) induced by lipopolysaccaride (LPS).Methods The acute lung injury rat model was induced by receiving 5 mg/kg LPS injection in tail vein.A total of 63 Wistar rats were divide into 3 groups without caring sex.Saline control group,LPS treated group and Ginaton treated group.The samples of different groups were collected 2 h,6 h,and 10 h after tail vein administration.Serum soluble cell ashesion molecule-1 (sICAM-1) was measured by ELISA,immunohistochemical staining and Western blotting of NF-kB were performed on sections of lung specimens.Results The acute lung injury rat model was successfully induced by receiving LPS injection in tail vein.The sICAM-1 levels increased more in Ginaton treated group than those in Saline control group (P＜0.01),and the increase in LPS treated group were the highest.By immunohistochemical staining,the positive NF-kB cells in Ginaton treated group were much less than those in LPS treated group (P＜0.01),and in Saline control group were least (P＜0.01).The results of the Western-blot method were consistent with immunohistochemical method.Conclusion ALI could be induced by LPS,Ginaton showed a protective effect through probably inhibiting activation of NF-kB on LPS-induced-ALI in this animal model.
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Objective To study the relationship among apoptosis,NF-KB activation and uPA expres- sion in human colon carcinoma cell line HCTll6 induced by 5-fluorouracil,and to observe the effect of in- hibiting activity of NF-KB by PDTC on apoptosis as well as expression of uPA.Methods Cell apoptosis was analysed by Annexin V-FITC.Fluctuation of NF-KB and uPA was detected by semi-quantitative immuno- histochemistry.Results 5-fluorouracil could induce apoptosis and activate NF-KB.PDTC could significantly increase the apoptosis and suppress the activation of NF-KB induced by 5-fluorouracil.There was a positive correlation between the changes of uPA and NF-KB.Conclusion 5-fluorouracil could induce apoptosis,ac- tivate NF-KB and up-regulate expression of uPA of HCT116 cells.The mechanism of enhanced apoptosis by PDTC may be related to suppressing activation of NF-?B and down-regulating expression of uPA.
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Objective To investigate the effect of PARP inhibitor 5-AIQ on PARP/NF-?B P65 complex and the NF-?B activity in murine colon carcinoma CT26 cells.Methods The murine colon carcinoma CT26 cells were treated with 5-AIQ in vitro.The expressions of PARP and NF-?B P65 were analyzed by Western Blot.The PARP/NF-?B P65 complex was analyzed by co-immunoprecipitation.The NF-?B binding activity was detected by electrophoretic mobility shift assay(EMSA).Results Compared with 5-AIQ-untreated group,the expressions of PARP and NF-?B P65 were markedly decreased in 5-AIQ-treated colon carcinoma CT26 cell groups(P
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Objective To investigate cell proliferation and invasiveness of cervical cancer HeLa229 cell after knockdown of NF-?B signaling pathway by P65 siRNA.Methods RNA interference was employed for specific inhibition of the expression of P65.HeLa229 cell was divided into transfected group and untransfected group.Cell viability was detected by MTT after the HeLa229 cells were transfected with or without P65 siRNA for 24,48,72 h.The sensitivity to 5-Fu of the HeLa229 cell,transfected with or without P65 siRNA,was evaluated also by MTT.Boyden chamber experiment in vitro was used to detect the invasion of HeLa229 cell.Results P65 siRNA inhibited the cell proliferation as compared with the untransfected cells.Proliferations of both cells transfected with and without P65 siRNA were inhibited in a concentration-dependent manner,while at the same concentration of 5-Fu the viability of transfected HeLa229 cells was significantly suppressed(P
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Objective To investigate the incidence of systemic inflammatory response syndrome(SIRS) after cardiopulmonary resuscitation(CPR) and to observe the effect of Ulinastain in inhibition of inflammatory mediator.Methods Forty patients surviving more than 48 hours after CPR were divided into Ulinastain and control groups randomly. Activity of nuclear factor kappa B(NF-?B), IL-6,TNF-? of the patients was detected .All patients were evaluated by SIRS diagnosis standard and their general organ function was examined. All data were compared between two groups.Results Activities of NF-?B, IL-6,TNF-? of patients after CPR was significantly higher than that of normal people (P