Objective To analyze the relationship between tumor necrosis factor-alpha (TNFα) gene promoter -308 polymorphism and myasthenia gravis (MG) in Chinese and analyze secretion of TNFα in peripheral blood mononuclear ce...Objective To analyze the relationship between tumor necrosis factor-alpha (TNFα) gene promoter -308 polymorphism and myasthenia gravis (MG) in Chinese and analyze secretion of TNFα in peripheral blood mononuclear cells (PBMC) in MG patients. Methods A biallelic polymorphism at position -308 in the promoter of TNFα gene was screened by PCR amplification and NcoI recognition site. One hundred and twenty-three MG cases and 115 healthy controls were included in this study. MG patients were classified to different groups according to clinical type, age at onset, and sex respectively. PBMC were isolated from 20 patients and 20 healthy controls, and then cultured in the presence or absence of phytohemag- glutinin (PHA) and acetycholine receptors (AchR). The supernatants were harvested after incubation and stored until TNFα was assayed by enzyme-linked immunosorbent assay. Results The frequency of TNFα-308 allele 2 (A) was found significantly increase in MG patients and showed a trend especially in late onset (≥ 40 years) and male patients (P < 0.05). The allele A had no relationship with thymic pathogenesis in MG patients. But frequency of allele A was significantly higher in general type than in ocular type (P < 0.05). MG patients had a higher inducible level of TNFα by PHA and AchR, and could be down regulated after treatment. Conclusion Polymorphism in TNFα gene promoter -308 is associated with onset of MG. The microsatellite allele TNFα2 confer risk for the development of MG in Chinese patients. MG patients have a higher inducible level of TNFα.展开更多
Objective: To explore the levels of serum and ascitic fluid soluble tumor necrosis factor receptor-p55 (sTNFR-p55) and understand their clinical implication in primary hepatocellular carcinoma (HCC) patients. Methods:...Objective: To explore the levels of serum and ascitic fluid soluble tumor necrosis factor receptor-p55 (sTNFR-p55) and understand their clinical implication in primary hepatocellular carcinoma (HCC) patients. Methods: Enzyme-linked immunosorbent assay (ELISA) was used to examine the levels of sTNFR-p55 in the serum and ascitic fluid in 25 HCC patients and 25 patients with liver cirrhosis (LC). The test was also performed on the serum of 30 healthy subjects who served as control group. To assess the clinical effects of increased serum concentrations of sTNFR-p55, four parameters were analyzed by logistic regression. Results: Serum and ascitic fluid levels of sTNFR-p55 in HCC patients were significantly higher than those in LC patients and controls (P=0. 001). No significant difference was found between serum sTNFR-p55 levels in the latter 2 groups (P = 0. 19), and positive correlation between serum levels of sTNFR-p55 and that in ascitic fluid was noted in the 2 patient groups (r=1. 000, P<0. 001). Levels of the sTNFR-p55 positively correlated with TBIL and AFP in the peripheral blood of HCC patients (r=0. 524, P = 0. 01 and r=0. 234, P = 0. 03, respectively). Conclusion: Increased levels of sTNFRs-p55 in the serum and ascitic fluid could reflect the abnormal immune status of the HCC patients and may help predict the development of the tumor.展开更多
Acute myocardial infarction （AMI） is an acute cardiovascular emergency. This study was undertaken to assess the effect of tumor necrosis factor-a （TNF-a） on ventricular arrhythmias induced byAMI in rats in vivo. ...Acute myocardial infarction （AMI） is an acute cardiovascular emergency. This study was undertaken to assess the effect of tumor necrosis factor-a （TNF-a） on ventricular arrhythmias induced byAMI in rats in vivo. Two hundred and forty male Wistar rats were randomized into a sham- operation group, an AMI group, and a recombinant human tumor necrosis factor receptor：Fc fusion protein（rhTNFR：Fc） group. Acute anterior wall myocardial infarction was produced in the AMI group by ligating the left anterior descending coronary artery （LAD）, and there was no ligation but operation in the sham-operation group. The rhTNFR：Fc group was treated with rhTNFR：Fc（10 mg/kg）, a TNF-a antagonist, 24 hours before LAD ligation. The spontaneous and induced programmed electrical stimulation ventricular arrhythmias were recorded at baseline and 10 minutes, 20 minutes, 30 minutes, 60 minutes, 3 hours, 6 hours and 12 hours after ligation. At the same time the protein and mRNA expression levels of TNF-a among different groups were detected by histochemistry and real-time fluorescent quantitative PCR. Expression of TNF-a increased markedly from 10 minutes after infarction, peaked at 20-30 minutes, and returned to baseline gradually in the AMI group and rhTNFR：Fc group. The time- windows of spontaneous and induced ventricular arrhythmias were similar. Compared with the AMI group, the rhTNFR：Fc group showed a lesser expression of TNF-a protein and a lower incidence of ventricular arrhythmias （P〈0.05）. There was no obvious change in the sham-operation group. The expression of TNF-a induced by AMI could contribute to the onset of ventricular arrhythmias.展开更多
In order to assess the role of tumor necrosis factor (TNF) in neonatal sepsis, plasma TNF levels were determined by a method using L929 cells at the time of septic work-up in 67 neonates. Thirty-three patients, with s...In order to assess the role of tumor necrosis factor (TNF) in neonatal sepsis, plasma TNF levels were determined by a method using L929 cells at the time of septic work-up in 67 neonates. Thirty-three patients, with sepsis were found to have significantly higher TNF levels (533. 33 ±468. 74 U/ml ; 1 U corresponding to 1. 67 pg recombinant TNF) as compared with 34 non-sepsis patients (100. 0±188, 97 U/ml) and 30 healthy newborns (27. 33±1 6. 1 7 U/ml, P<0. 05. respectively) . The upper limit of normal plasma TNF levels was 60 U/ml and the best cutoff value for predicting neonatal sepsis was 160 U/ml. This had remarkable sensitivity (88%), specificity (82%). positive predictive value (83%). and negative predictive value (88%). Plasma TNF levels were significantly associated with the occurrence of shock,organ failure. sclerema and outcome. Thus, anti-TNF antibodies might be used in protecting newborns from septic death.展开更多
Objective.In this study,we investigated the hypothesis that tumor necrosis factor(TNF)α-308gene polymorphism might be of the genetic predisposition to asthma and asthma phenotypes.Methods.TNFα-308gene polymorphism w...Objective.In this study,we investigated the hypothesis that tumor necrosis factor(TNF)α-308gene polymorphism might be of the genetic predisposition to asthma and asthma phenotypes.Methods.TNFα-308gene polymorphism was genotyped in221random unrelated Northern Chinese population(comprising125asthmatics and96healthy controls)and52individuals from12asthmatic families with Han ethnic by using polymerase chain reaction(PCR)-restriction fragment length polymor-phism(RFLP).Methacholine(Mch)broncho-challenge test,bronchial reversibility test and lung function were underwent in all asthmatics.Results.TNFα-3082homozygosity was present at a significantly higher frequency in asthmatics than that in controls(20.8%vs11.4%,P<0.05,OR2.259),the TNF allele2was also higher in asthmatics compared with controls(0.42vs0.33,P<0.01).TNFα-3082homozygosity was an weak independent risk factor for asthma etiology(OR0.226,P<0.05).Moreover,patients carrying TNFα-3082homozy-gosity had less responsive to inhaledβ 2 -agonist in20minutes than patients carrying other two genotypes(24.1%vs29.5%vs38.8%,P<0.05).Linkage analysis didn’t support that TNFαgene was linked to asthma (Likelihood of odds,LOD<1)based on familial data.Conclusion These results suggest that TNFα-3082homozygosity may be of a component contribut-ing to the genetic predisposition to asthma ,and airway responsiveness toβ2 -agonist.展开更多
Objective: To investigate membrane tumor necrosis factor receptor 1 protein expression level in decidua and concentration of soluble tumor necrosis factor receptor 1 in serum in women with unexplained early spontaneou...Objective: To investigate membrane tumor necrosis factor receptor 1 protein expression level in decidua and concentration of soluble tumor necrosis factor receptor 1 in serum in women with unexplained early spontaneous abortion, threatened abortion, and compare the levels with healthy pregnant women. Methods: Thirty-seven women with unexplained early spontaneous abortion, 27 women with threatened abortion, and 34 healthy pregnant women undergoing artificial abortion of pregnancy at 6 - 10 weeks of gestation were selected. Decidual samples were collected when women were undergoing artificial abortion, and blood samples were collected at the same time. The level of membrane tumor necrosis factor receptor 1 in decidua was detected by flow cytometer, and the concentration of soluble tumor necrosis factor receptor 1 in sera was measured with an enzyme-linked immunosorbent assay. Results: The percentages of membrane tumor necrosis factor receptor 1 positive decidual cells were 16.42 ± 7.10 Mean ± SD for women with unexplained early spontaneous abortion and 13. 14 ± 6.30 for healthy pregnant women ( P < 0.05). Serum concentration of soluble tumor necrosis factor receptor 1 was significantly higher in women with unexplained early spontaneous abortion than in healthy pregnant women and in women with threatened abortion, and no difference was found between healthy pregnant women and women with threatened abortion. Conclusion: Women with unexplained early spontaneous abortion present significantly higher expression of tumor necrosis factor receptor 1 than healthy pregnant women, suggesting that over-expression of tumor necrosis factor receptor 1 may contribute to the development of early spontaneous abortion.展开更多
Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and it...Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms. Methods The expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFNγ, TRAIL, IFNγ ＋ TRAIL, IFNγ ＋ Caspase 8 inhibitor ＋ TRAIL, IFNγ ＋ cisplatin ＋ TRAIL, and IFNγ ＋ etoposide ＋ TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay. Results Caspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNγ. SH-SY5Y ceils themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNγ were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFNγ ＋ TRAIL group was significantly higher than those of control group, IFNγ group, TRAIL group, and inhibitor group （ P 〈 0. 01 ）. There was no significant difference among IFNγ ＋ TRAIL group, IFNγ ＋ cisplatin ＋ TRAIL group, and IFNγ ＋ etoposide ＋ TRAIL group. Conclusions IFNγ could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the up-regulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.展开更多
BACKGROUND： The tumor necrosis factor recepter associated factor （TRAF） 6 is an important intracellular adapter protein that plays a pivotal role in activating multiple inflammatory and immune related processes ind...BACKGROUND： The tumor necrosis factor recepter associated factor （TRAF） 6 is an important intracellular adapter protein that plays a pivotal role in activating multiple inflammatory and immune related processes induced by cytokines. TRAF6 represents a strong candidate susceptibility factor for sepsis. We investigated whether polymorphisms at the TRAF6 gene are associated with the susceptibility to and severity of sepsis.METHODS： A hospital-based case-control study was conducted with 255 patients with sepsis and 260 controls who were recruited from Zhengzhou, China. Haplotype tagging single nucleotide polymorphisms （htSNPs） were selected from the HapMap database and genotyped using the SNPstream genotyping platform. The associations with the susceptibility and disease severity of sepsis were estimated by logistic regression, and adjusted for age, sex, smoking, drinking, chronic diseases status, APACHEII score and critical illness status.RESULTS： A total of 13 TRAF6 SNPs were tagged by 7 htSNPs. Five htSNPs （rs5030490, rs5030411, rs5030416, rs5030445 and rs3740961） were genotyped in the case control study. Genotype frequencies of the htSNPs were conformed to the Hardy-Weinberg equilibrium in both patients and controls. No significant association was found between the 5 htSNPs and the susceptibility to and severity of sepsis. Compared with the main haplotype -11120A/-10688T/-9423A/805G/12967G, no certain haplotype was associated with the signi？ cantly susceptibility to or severity of sepsis.CONCLUSION： TRAF6 gene polymorphisms might not play a major role in mediating the susceptibility to and severity of sepsis in the Chinese population. A larger population-based case-control study is warranted.展开更多
Objective: To study the molecular mechanisms of TNF--a expression induced by lipopolysaccharide (LPS) for exploring novel methods to prevent or treat clinical patients with endotoxic shock. Methods:Protein kinase assa...Objective: To study the molecular mechanisms of TNF--a expression induced by lipopolysaccharide (LPS) for exploring novel methods to prevent or treat clinical patients with endotoxic shock. Methods:Protein kinase assay was used to detect the kinase activity stimulated by LPS; Con focal laser scan technique was used to show the translocation of p38 on the activation; RT PCR and reporter gene system were used to study the molecular mechanism of TNF -a gene transcription. Results: In RAW cells it was found that p38 was activated on the stimulation of LPS. and activated p38 moved into nucleus from cytosol; TNF--a mRNA increased on the stimulation of LPS and the increased promoter transactivity induced by LPS could be inhibited significantly by specific inhibitor for p38. Conclusion: p38 mitogen activated protein kinase (MAPK ) was activated by the stimulation of LPS,which brought about its entry to the nucleus to act on transcription factors to regulate cellular processes. p38 MAPK Is an important regulator of TNF--a gene expression induced by LPS.展开更多
Background Cilostazol, an anti-platelet drug for treating coronary heart disease, has been reported to modulate immune cell functions Plasmacytoid dendritic cells （pDCs） have been found to participate in the progres...Background Cilostazol, an anti-platelet drug for treating coronary heart disease, has been reported to modulate immune cell functions Plasmacytoid dendritic cells （pDCs） have been found to participate in the progression of atherosclerosis mainly through interferon ct （IFN-ct） production. Whether cilostazol influences pDCs activation is still not clear. In this study, we aimed to investigate the effects of cilostazol on cell activation and antigen presentation ofpDCs in vitro in this study. Methods Peripheral blood mononuclear cells isolated by Ficoll cen- trifugation and pDCs sorted by flow cytometry were used in this study. After pretreated with cilostazol for 2 h, cells were stimulated with CpG-A, R848 or virus for 6 h or 20 h, or stimulated with CpG-B for 48 h and then co-cultured with naive T cell for five days. Cytokines in supernatant and intracellular cytokines were analyzed by ELISA or flow cytometry respectively. Results Our data indicated that cilostazol could inhibit IFN-α and tumor necrosis factor α （TNF-α） production from pDCs in a dose-dependent manner. In addition, the ability of priming na ve T cells of pDCs was also impaired by cilostazol. The inhibitory effect was not due to cell killing since the viability of pDCs did not change upon cilostazol treatment. Conclusion Cilostazol inhibits pDCs cell activation and antigen presentation in vitro, which may explain how cilostazol protects against atherosclerosis.展开更多
Objective: To study the role of p38MAPK in mediating TNF-α-induced apoptosis of rat glioma cell line C6. Methods: Effect of TNF-α on the proliferation of C6 cells was determined by MTT assay. The TNF-α induced apop...Objective: To study the role of p38MAPK in mediating TNF-α-induced apoptosis of rat glioma cell line C6. Methods: Effect of TNF-α on the proliferation of C6 cells was determined by MTT assay. The TNF-α induced apoptosis was detected by transmission electron microscopy and flow cytometry. The expression of p38MAPK was detected by SABC method and Western-blot. The effect of SB202190, a specific inhibitor of p38MAPK, on TNF-α-induced apoptosis was observed by flow cytometry and SABC method. Results: Inhibitory rate of TNF-α(2×105 U/L) on C6 cells was 43.75%. In the TNF-α treated group, apoptotic cells were observed by transmission electron microscopy and the apoptotic rate was 37.5% by flow cytometry. p38MAPK positive signals were detected by SABC method and Western-blot. In the SB202190 treated group, the apoptotic rate was 7.0% and no p38MAPK signals were found. Conclusion: Apoptosis of C6 cells and expression of p38MAPK can be induced by TNF-α. The activation of p38MAPK promotes the apoptosis of C6 cells.展开更多
Objective： To study whether the anti-AS effect of ginkgo biloba extract （GbE） was related with inhibitory effects on the expression of IL-1β, TNF-α and up-regulation of IL-10 and IL-10R in the heart of atheroscle...Objective： To study whether the anti-AS effect of ginkgo biloba extract （GbE） was related with inhibitory effects on the expression of IL-1β, TNF-α and up-regulation of IL-10 and IL-10R in the heart of atherosclerotic （AS） rats. Methods： The experimental model of AS rats were established by intraperitioneal injection of vitamin D3 with high fat and cholesterol diet. All rats were divided into 3 groups： control, AS and GbE. GbE （100 mg/kg） was administered to rats by ig. After 8 weeks, the expression of IL-1β, TNF-α, IL-10, and IL-10R in the heart of AS rats were detected by enzyme-linked immunosorbant assay, reverse transcriptase polymerasechain reaction and Western blotting. Results： The protein and mRNA expressions of IL-1β, TNF-α, IL-10 and IL-10R were markedly higher in AS group than those in control group （P〈0.01）. The protein and mRNA expressions of IL-1β and TNF-α were markedly lower in GbE group than those in AS group; while the protein and mRNA expressions of IL-10 and IL-10R were markedly higher in GbE group than those in AS group （P〈0.01）. Conclusion： GbE has significant inhibitory effects on proinflammatory cytokine IL-1β, TNF-α. The up-regulation of anti-inflammatory cytokine IL-10, IL-10R that may he partially responsible for its anti-AS effects.展开更多
Objective: To observe the changes of plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin in children undergoing cardiac procedure and to st...Objective: To observe the changes of plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin in children undergoing cardiac procedure and to study the effects of cardiopulmonary bypass (CPB) on the injury or activation of endothelial cells and vascular permeability. Methods: Twenty children undergoing cardiac operation with CPB were selected in the study. Plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin were measured after anesthetic induction (baseline), bypass for 20 minutes, at the end of CPB, and at 2, 4, and 18 h after the end of CPB. Results: The plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, and urinary microalbumin began to increase at 2 h after the end of CPB, and remained higher than that of the baseline, while the concentration of tumor necrosis factor α increased only at the end CPB and at 2 h after the end of CPB. Conclusion: Cardiopulmonary bypass can induce inflammatory response, resulting in the activation or injury of vascular endothelial cells, and can increase the vascular permeability.展开更多
Objective：To evaluate the effects of beating-heart and arrested heart intracardiac procedure on the expression of tumor necrosis factor alpha （TNF-α mRNA in myocardium. Methods： Thirty congenital ventricular septa...Objective：To evaluate the effects of beating-heart and arrested heart intracardiac procedure on the expression of tumor necrosis factor alpha （TNF-α mRNA in myocardium. Methods： Thirty congenital ventricular septal defect （VSD） patients aged from 5 to 10 years old were randomly divided into 2 groups equally. Group A underwent traditional arrested heart intracardiac procedures ; group B underwent beating-heart procedures. Specimens of myocardium were obtained at the onset （baseline） and the end of cardiopulmonary bypass （CPB） for the determination of TNF a mRNA. Concentration of TNF-α was respectively measured after anesthetic induction （T1）, 20 min after the beginning of CPB （T2）, at the end of CPB （T3） and 6, 12, 24 h after CPB （T4-6） in all patierits： After separating polymorphonuclear leucocyte （PMN）, we distilled nuclear protein and mensurated the activation of nuclear factor-κB （NF-κB） by elec-trophoretic mobility shift assay （EMSA）. Results ：Compared with baseline, the expression of TNF-κ mRNA significantly increased in both groups （P〈0. 05）. TNF-α mRNA level of group A was significantly higher than that of group B at the end of CPB （P〈0.05）. The plasma concentration of TNF-α and neutrophil NF-κB activity in group A was significantly higher than that of group B at T,4-6（P〈0.05）. Conclusion：Compared with traditional arrested CPB, beating heart intracadiac procedure can effectively reduce the expression and release of TNF-α; it will benefit the protection of pediatric myocardial during CPB.展开更多
A 27 year-old male patient was diagnosed with Behcet＇s disease cortex）, （TNF） which resulted in reduced visual acuity and field of vision. This alpha blocker therapy on Behcet＇s cerebral vasculitis （BD） case a...A 27 year-old male patient was diagnosed with Behcet＇s disease cortex）, （TNF） which resulted in reduced visual acuity and field of vision. This alpha blocker therapy on Behcet＇s cerebral vasculitis （BD） case and posterior visual pathway demonstrates the effect of tum affection （visual or necrosis factor展开更多
The present study was designed to investigate the production of tumor necrosis factor a (TNFα) in-duced by low-dose (1μg/kg) lipopolysaccharide (LPS) and its cellular source after hemorrhagic shock(HS) in rats, and ...The present study was designed to investigate the production of tumor necrosis factor a (TNFα) in-duced by low-dose (1μg/kg) lipopolysaccharide (LPS) and its cellular source after hemorrhagic shock(HS) in rats, and to further analyze the mechanism for increased sensitivity to LPS through looking at ex-pression of lipopolysaccharide -binding protein (LBP ) mRN A in t he liver, lungs and kidneys. lt wa s foundin uiuo that plasma TNFa levels in the HS+LPS group were 20-fold higher than that in the HS group (P<0. 01 ), and 2. 7-fold higher than that in the LPS group (P<0. 05). lt was shown in ndro that the ca-pacity of peripheral white blood cells to produce TNFa in response to LPS stimulation was significantly de-creased by 126 % (P<0. 01 ) and 57% (P<0. 05) compared with pre-shock levels and the sham group re-spectively at the end of resuscitation following shock, and was still markedly decreased 3 hours after resus-citation, while the capacity of Kupffer Cells was significantly increased by 110% compared with the shamgroup (P<0. 01) after shock and resuscitation. Results from RT-PCR showed that expression of LBPmRNA in the liver, lungs and kidneys was increased after shock and resuscitation. It is suggested thathemorrhagic shock could significantly enhance endotoxin-induced TNFa production, which might be due toup-regulation of LBP expression in tissues after shock, and tissue macrophages might be the main source ofcytokine production.展开更多
Fifteen hybridoma cell lines producing monoclonal antiboclies (McAb) against recombinant human tu-mor necrosis factor a (rhTNFa) have been established by fusing SP 2/0 cells with spleen cells from aBALB/c mouse immuni...Fifteen hybridoma cell lines producing monoclonal antiboclies (McAb) against recombinant human tu-mor necrosis factor a (rhTNFa) have been established by fusing SP 2/0 cells with spleen cells from aBALB/c mouse immunized with rhTNFa. Following J M Davis’s Works, semi-solid medium was usedfor initial cloning. Five of them were studied further. Their main chromosome- numbers range were 96 to105, all of them were IgG1 subclass. The affinities of these McAbs were estimated to be 1. 25 ×108 mol/L, 1. 12×108 mol/L, 2. 34×108 mol/L, 8. 55 × 107 mol/L, 1. 04×108 mol/L, respectively.Two groups of mice challenging with E Coli (107 organisms), one group treated with 2mg/kg anti-TNF monoclonal antibody, the other did not. There was a higher survival rate in treated group, the serumTNF level was significantly lower too, and the untreated mice had severe pathologic changes in vlscera.展开更多
文摘Objective To analyze the relationship between tumor necrosis factor-alpha (TNFα) gene promoter -308 polymorphism and myasthenia gravis (MG) in Chinese and analyze secretion of TNFα in peripheral blood mononuclear cells (PBMC) in MG patients. Methods A biallelic polymorphism at position -308 in the promoter of TNFα gene was screened by PCR amplification and NcoI recognition site. One hundred and twenty-three MG cases and 115 healthy controls were included in this study. MG patients were classified to different groups according to clinical type, age at onset, and sex respectively. PBMC were isolated from 20 patients and 20 healthy controls, and then cultured in the presence or absence of phytohemag- glutinin (PHA) and acetycholine receptors (AchR). The supernatants were harvested after incubation and stored until TNFα was assayed by enzyme-linked immunosorbent assay. Results The frequency of TNFα-308 allele 2 (A) was found significantly increase in MG patients and showed a trend especially in late onset (≥ 40 years) and male patients (P < 0.05). The allele A had no relationship with thymic pathogenesis in MG patients. But frequency of allele A was significantly higher in general type than in ocular type (P < 0.05). MG patients had a higher inducible level of TNFα by PHA and AchR, and could be down regulated after treatment. Conclusion Polymorphism in TNFα gene promoter -308 is associated with onset of MG. The microsatellite allele TNFα2 confer risk for the development of MG in Chinese patients. MG patients have a higher inducible level of TNFα.
文摘Objective: To explore the levels of serum and ascitic fluid soluble tumor necrosis factor receptor-p55 (sTNFR-p55) and understand their clinical implication in primary hepatocellular carcinoma (HCC) patients. Methods: Enzyme-linked immunosorbent assay (ELISA) was used to examine the levels of sTNFR-p55 in the serum and ascitic fluid in 25 HCC patients and 25 patients with liver cirrhosis (LC). The test was also performed on the serum of 30 healthy subjects who served as control group. To assess the clinical effects of increased serum concentrations of sTNFR-p55, four parameters were analyzed by logistic regression. Results: Serum and ascitic fluid levels of sTNFR-p55 in HCC patients were significantly higher than those in LC patients and controls (P=0. 001). No significant difference was found between serum sTNFR-p55 levels in the latter 2 groups (P = 0. 19), and positive correlation between serum levels of sTNFR-p55 and that in ascitic fluid was noted in the 2 patient groups (r=1. 000, P<0. 001). Levels of the sTNFR-p55 positively correlated with TBIL and AFP in the peripheral blood of HCC patients (r=0. 524, P = 0. 01 and r=0. 234, P = 0. 03, respectively). Conclusion: Increased levels of sTNFRs-p55 in the serum and ascitic fluid could reflect the abnormal immune status of the HCC patients and may help predict the development of the tumor.
文摘Acute myocardial infarction （AMI） is an acute cardiovascular emergency. This study was undertaken to assess the effect of tumor necrosis factor-a （TNF-a） on ventricular arrhythmias induced byAMI in rats in vivo. Two hundred and forty male Wistar rats were randomized into a sham- operation group, an AMI group, and a recombinant human tumor necrosis factor receptor：Fc fusion protein（rhTNFR：Fc） group. Acute anterior wall myocardial infarction was produced in the AMI group by ligating the left anterior descending coronary artery （LAD）, and there was no ligation but operation in the sham-operation group. The rhTNFR：Fc group was treated with rhTNFR：Fc（10 mg/kg）, a TNF-a antagonist, 24 hours before LAD ligation. The spontaneous and induced programmed electrical stimulation ventricular arrhythmias were recorded at baseline and 10 minutes, 20 minutes, 30 minutes, 60 minutes, 3 hours, 6 hours and 12 hours after ligation. At the same time the protein and mRNA expression levels of TNF-a among different groups were detected by histochemistry and real-time fluorescent quantitative PCR. Expression of TNF-a increased markedly from 10 minutes after infarction, peaked at 20-30 minutes, and returned to baseline gradually in the AMI group and rhTNFR：Fc group. The time- windows of spontaneous and induced ventricular arrhythmias were similar. Compared with the AMI group, the rhTNFR：Fc group showed a lesser expression of TNF-a protein and a lower incidence of ventricular arrhythmias （P〈0.05）. There was no obvious change in the sham-operation group. The expression of TNF-a induced by AMI could contribute to the onset of ventricular arrhythmias.
文摘In order to assess the role of tumor necrosis factor (TNF) in neonatal sepsis, plasma TNF levels were determined by a method using L929 cells at the time of septic work-up in 67 neonates. Thirty-three patients, with sepsis were found to have significantly higher TNF levels (533. 33 ±468. 74 U/ml ; 1 U corresponding to 1. 67 pg recombinant TNF) as compared with 34 non-sepsis patients (100. 0±188, 97 U/ml) and 30 healthy newborns (27. 33±1 6. 1 7 U/ml, P<0. 05. respectively) . The upper limit of normal plasma TNF levels was 60 U/ml and the best cutoff value for predicting neonatal sepsis was 160 U/ml. This had remarkable sensitivity (88%), specificity (82%). positive predictive value (83%). and negative predictive value (88%). Plasma TNF levels were significantly associated with the occurrence of shock,organ failure. sclerema and outcome. Thus, anti-TNF antibodies might be used in protecting newborns from septic death.
文摘Objective.In this study,we investigated the hypothesis that tumor necrosis factor(TNF)α-308gene polymorphism might be of the genetic predisposition to asthma and asthma phenotypes.Methods.TNFα-308gene polymorphism was genotyped in221random unrelated Northern Chinese population(comprising125asthmatics and96healthy controls)and52individuals from12asthmatic families with Han ethnic by using polymerase chain reaction(PCR)-restriction fragment length polymor-phism(RFLP).Methacholine(Mch)broncho-challenge test,bronchial reversibility test and lung function were underwent in all asthmatics.Results.TNFα-3082homozygosity was present at a significantly higher frequency in asthmatics than that in controls(20.8%vs11.4%,P<0.05,OR2.259),the TNF allele2was also higher in asthmatics compared with controls(0.42vs0.33,P<0.01).TNFα-3082homozygosity was an weak independent risk factor for asthma etiology(OR0.226,P<0.05).Moreover,patients carrying TNFα-3082homozy-gosity had less responsive to inhaledβ 2 -agonist in20minutes than patients carrying other two genotypes(24.1%vs29.5%vs38.8%,P<0.05).Linkage analysis didn’t support that TNFαgene was linked to asthma (Likelihood of odds,LOD<1)based on familial data.Conclusion These results suggest that TNFα-3082homozygosity may be of a component contribut-ing to the genetic predisposition to asthma ,and airway responsiveness toβ2 -agonist.
文摘Objective: To investigate membrane tumor necrosis factor receptor 1 protein expression level in decidua and concentration of soluble tumor necrosis factor receptor 1 in serum in women with unexplained early spontaneous abortion, threatened abortion, and compare the levels with healthy pregnant women. Methods: Thirty-seven women with unexplained early spontaneous abortion, 27 women with threatened abortion, and 34 healthy pregnant women undergoing artificial abortion of pregnancy at 6 - 10 weeks of gestation were selected. Decidual samples were collected when women were undergoing artificial abortion, and blood samples were collected at the same time. The level of membrane tumor necrosis factor receptor 1 in decidua was detected by flow cytometer, and the concentration of soluble tumor necrosis factor receptor 1 in sera was measured with an enzyme-linked immunosorbent assay. Results: The percentages of membrane tumor necrosis factor receptor 1 positive decidual cells were 16.42 ± 7.10 Mean ± SD for women with unexplained early spontaneous abortion and 13. 14 ± 6.30 for healthy pregnant women ( P < 0.05). Serum concentration of soluble tumor necrosis factor receptor 1 was significantly higher in women with unexplained early spontaneous abortion than in healthy pregnant women and in women with threatened abortion, and no difference was found between healthy pregnant women and women with threatened abortion. Conclusion: Women with unexplained early spontaneous abortion present significantly higher expression of tumor necrosis factor receptor 1 than healthy pregnant women, suggesting that over-expression of tumor necrosis factor receptor 1 may contribute to the development of early spontaneous abortion.
基金the National Natural Sciences Foundation of China(39470739)the Ministry of Public Health Research Foundation(20122167)the Doctor Startup-Natural Science Foundation of Li-aoning Province (20041047)
文摘Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms. Methods The expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFNγ, TRAIL, IFNγ ＋ TRAIL, IFNγ ＋ Caspase 8 inhibitor ＋ TRAIL, IFNγ ＋ cisplatin ＋ TRAIL, and IFNγ ＋ etoposide ＋ TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay. Results Caspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNγ. SH-SY5Y ceils themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNγ were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFNγ ＋ TRAIL group was significantly higher than those of control group, IFNγ group, TRAIL group, and inhibitor group （ P 〈 0. 01 ）. There was no significant difference among IFNγ ＋ TRAIL group, IFNγ ＋ cisplatin ＋ TRAIL group, and IFNγ ＋ etoposide ＋ TRAIL group. Conclusions IFNγ could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the up-regulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.
文摘BACKGROUND： The tumor necrosis factor recepter associated factor （TRAF） 6 is an important intracellular adapter protein that plays a pivotal role in activating multiple inflammatory and immune related processes induced by cytokines. TRAF6 represents a strong candidate susceptibility factor for sepsis. We investigated whether polymorphisms at the TRAF6 gene are associated with the susceptibility to and severity of sepsis.METHODS： A hospital-based case-control study was conducted with 255 patients with sepsis and 260 controls who were recruited from Zhengzhou, China. Haplotype tagging single nucleotide polymorphisms （htSNPs） were selected from the HapMap database and genotyped using the SNPstream genotyping platform. The associations with the susceptibility and disease severity of sepsis were estimated by logistic regression, and adjusted for age, sex, smoking, drinking, chronic diseases status, APACHEII score and critical illness status.RESULTS： A total of 13 TRAF6 SNPs were tagged by 7 htSNPs. Five htSNPs （rs5030490, rs5030411, rs5030416, rs5030445 and rs3740961） were genotyped in the case control study. Genotype frequencies of the htSNPs were conformed to the Hardy-Weinberg equilibrium in both patients and controls. No significant association was found between the 5 htSNPs and the susceptibility to and severity of sepsis. Compared with the main haplotype -11120A/-10688T/-9423A/805G/12967G, no certain haplotype was associated with the signi？ cantly susceptibility to or severity of sepsis.CONCLUSION： TRAF6 gene polymorphisms might not play a major role in mediating the susceptibility to and severity of sepsis in the Chinese population. A larger population-based case-control study is warranted.
文摘Objective: To study the molecular mechanisms of TNF--a expression induced by lipopolysaccharide (LPS) for exploring novel methods to prevent or treat clinical patients with endotoxic shock. Methods:Protein kinase assay was used to detect the kinase activity stimulated by LPS; Con focal laser scan technique was used to show the translocation of p38 on the activation; RT PCR and reporter gene system were used to study the molecular mechanism of TNF -a gene transcription. Results: In RAW cells it was found that p38 was activated on the stimulation of LPS. and activated p38 moved into nucleus from cytosol; TNF--a mRNA increased on the stimulation of LPS and the increased promoter transactivity induced by LPS could be inhibited significantly by specific inhibitor for p38. Conclusion: p38 mitogen activated protein kinase (MAPK ) was activated by the stimulation of LPS,which brought about its entry to the nucleus to act on transcription factors to regulate cellular processes. p38 MAPK Is an important regulator of TNF--a gene expression induced by LPS.
文摘Background Cilostazol, an anti-platelet drug for treating coronary heart disease, has been reported to modulate immune cell functions Plasmacytoid dendritic cells （pDCs） have been found to participate in the progression of atherosclerosis mainly through interferon ct （IFN-ct） production. Whether cilostazol influences pDCs activation is still not clear. In this study, we aimed to investigate the effects of cilostazol on cell activation and antigen presentation ofpDCs in vitro in this study. Methods Peripheral blood mononuclear cells isolated by Ficoll cen- trifugation and pDCs sorted by flow cytometry were used in this study. After pretreated with cilostazol for 2 h, cells were stimulated with CpG-A, R848 or virus for 6 h or 20 h, or stimulated with CpG-B for 48 h and then co-cultured with naive T cell for five days. Cytokines in supernatant and intracellular cytokines were analyzed by ELISA or flow cytometry respectively. Results Our data indicated that cilostazol could inhibit IFN-α and tumor necrosis factor α （TNF-α） production from pDCs in a dose-dependent manner. In addition, the ability of priming na ve T cells of pDCs was also impaired by cilostazol. The inhibitory effect was not due to cell killing since the viability of pDCs did not change upon cilostazol treatment. Conclusion Cilostazol inhibits pDCs cell activation and antigen presentation in vitro, which may explain how cilostazol protects against atherosclerosis.
文摘Objective: To study the role of p38MAPK in mediating TNF-α-induced apoptosis of rat glioma cell line C6. Methods: Effect of TNF-α on the proliferation of C6 cells was determined by MTT assay. The TNF-α induced apoptosis was detected by transmission electron microscopy and flow cytometry. The expression of p38MAPK was detected by SABC method and Western-blot. The effect of SB202190, a specific inhibitor of p38MAPK, on TNF-α-induced apoptosis was observed by flow cytometry and SABC method. Results: Inhibitory rate of TNF-α(2×105 U/L) on C6 cells was 43.75%. In the TNF-α treated group, apoptotic cells were observed by transmission electron microscopy and the apoptotic rate was 37.5% by flow cytometry. p38MAPK positive signals were detected by SABC method and Western-blot. In the SB202190 treated group, the apoptotic rate was 7.0% and no p38MAPK signals were found. Conclusion: Apoptosis of C6 cells and expression of p38MAPK can be induced by TNF-α. The activation of p38MAPK promotes the apoptosis of C6 cells.
基金Supported by the National Natural Science Foundation of China (No. 30271509)
文摘Objective： To study whether the anti-AS effect of ginkgo biloba extract （GbE） was related with inhibitory effects on the expression of IL-1β, TNF-α and up-regulation of IL-10 and IL-10R in the heart of atherosclerotic （AS） rats. Methods： The experimental model of AS rats were established by intraperitioneal injection of vitamin D3 with high fat and cholesterol diet. All rats were divided into 3 groups： control, AS and GbE. GbE （100 mg/kg） was administered to rats by ig. After 8 weeks, the expression of IL-1β, TNF-α, IL-10, and IL-10R in the heart of AS rats were detected by enzyme-linked immunosorbant assay, reverse transcriptase polymerasechain reaction and Western blotting. Results： The protein and mRNA expressions of IL-1β, TNF-α, IL-10 and IL-10R were markedly higher in AS group than those in control group （P〈0.01）. The protein and mRNA expressions of IL-1β and TNF-α were markedly lower in GbE group than those in AS group; while the protein and mRNA expressions of IL-10 and IL-10R were markedly higher in GbE group than those in AS group （P〈0.01）. Conclusion： GbE has significant inhibitory effects on proinflammatory cytokine IL-1β, TNF-α. The up-regulation of anti-inflammatory cytokine IL-10, IL-10R that may he partially responsible for its anti-AS effects.
文摘Objective: To observe the changes of plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin in children undergoing cardiac procedure and to study the effects of cardiopulmonary bypass (CPB) on the injury or activation of endothelial cells and vascular permeability. Methods: Twenty children undergoing cardiac operation with CPB were selected in the study. Plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin were measured after anesthetic induction (baseline), bypass for 20 minutes, at the end of CPB, and at 2, 4, and 18 h after the end of CPB. Results: The plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, and urinary microalbumin began to increase at 2 h after the end of CPB, and remained higher than that of the baseline, while the concentration of tumor necrosis factor α increased only at the end CPB and at 2 h after the end of CPB. Conclusion: Cardiopulmonary bypass can induce inflammatory response, resulting in the activation or injury of vascular endothelial cells, and can increase the vascular permeability.
文摘Objective：To evaluate the effects of beating-heart and arrested heart intracardiac procedure on the expression of tumor necrosis factor alpha （TNF-α mRNA in myocardium. Methods： Thirty congenital ventricular septal defect （VSD） patients aged from 5 to 10 years old were randomly divided into 2 groups equally. Group A underwent traditional arrested heart intracardiac procedures ; group B underwent beating-heart procedures. Specimens of myocardium were obtained at the onset （baseline） and the end of cardiopulmonary bypass （CPB） for the determination of TNF a mRNA. Concentration of TNF-α was respectively measured after anesthetic induction （T1）, 20 min after the beginning of CPB （T2）, at the end of CPB （T3） and 6, 12, 24 h after CPB （T4-6） in all patierits： After separating polymorphonuclear leucocyte （PMN）, we distilled nuclear protein and mensurated the activation of nuclear factor-κB （NF-κB） by elec-trophoretic mobility shift assay （EMSA）. Results ：Compared with baseline, the expression of TNF-κ mRNA significantly increased in both groups （P〈0. 05）. TNF-α mRNA level of group A was significantly higher than that of group B at the end of CPB （P〈0.05）. The plasma concentration of TNF-α and neutrophil NF-κB activity in group A was significantly higher than that of group B at T,4-6（P〈0.05）. Conclusion：Compared with traditional arrested CPB, beating heart intracadiac procedure can effectively reduce the expression and release of TNF-α; it will benefit the protection of pediatric myocardial during CPB.
文摘A 27 year-old male patient was diagnosed with Behcet＇s disease cortex）, （TNF） which resulted in reduced visual acuity and field of vision. This alpha blocker therapy on Behcet＇s cerebral vasculitis （BD） case and posterior visual pathway demonstrates the effect of tum affection （visual or necrosis factor
文摘The present study was designed to investigate the production of tumor necrosis factor a (TNFα) in-duced by low-dose (1μg/kg) lipopolysaccharide (LPS) and its cellular source after hemorrhagic shock(HS) in rats, and to further analyze the mechanism for increased sensitivity to LPS through looking at ex-pression of lipopolysaccharide -binding protein (LBP ) mRN A in t he liver, lungs and kidneys. lt wa s foundin uiuo that plasma TNFa levels in the HS+LPS group were 20-fold higher than that in the HS group (P<0. 01 ), and 2. 7-fold higher than that in the LPS group (P<0. 05). lt was shown in ndro that the ca-pacity of peripheral white blood cells to produce TNFa in response to LPS stimulation was significantly de-creased by 126 % (P<0. 01 ) and 57% (P<0. 05) compared with pre-shock levels and the sham group re-spectively at the end of resuscitation following shock, and was still markedly decreased 3 hours after resus-citation, while the capacity of Kupffer Cells was significantly increased by 110% compared with the shamgroup (P<0. 01) after shock and resuscitation. Results from RT-PCR showed that expression of LBPmRNA in the liver, lungs and kidneys was increased after shock and resuscitation. It is suggested thathemorrhagic shock could significantly enhance endotoxin-induced TNFa production, which might be due toup-regulation of LBP expression in tissues after shock, and tissue macrophages might be the main source ofcytokine production.
文摘Fifteen hybridoma cell lines producing monoclonal antiboclies (McAb) against recombinant human tu-mor necrosis factor a (rhTNFa) have been established by fusing SP 2/0 cells with spleen cells from aBALB/c mouse immunized with rhTNFa. Following J M Davis’s Works, semi-solid medium was usedfor initial cloning. Five of them were studied further. Their main chromosome- numbers range were 96 to105, all of them were IgG1 subclass. The affinities of these McAbs were estimated to be 1. 25 ×108 mol/L, 1. 12×108 mol/L, 2. 34×108 mol/L, 8. 55 × 107 mol/L, 1. 04×108 mol/L, respectively.Two groups of mice challenging with E Coli (107 organisms), one group treated with 2mg/kg anti-TNF monoclonal antibody, the other did not. There was a higher survival rate in treated group, the serumTNF level was significantly lower too, and the untreated mice had severe pathologic changes in vlscera.
