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Objectives:To examine whether lipopolysaccharide (LPS) induced apoptosis correlates with TNF ? release by type Ⅱ alveolar epithelial cells (AEC Ⅱ), whether TNF ? knockout (TNF KO) abrogates the induction of apoptosis by LPS and whether TNF ? is sufficient to induce apoptosis in this cell type. Methods:AEC Ⅱ was isolated from wild type mice and TNF KO mice. Cells were stimulated with LPS or recombinant murine TNF ? for 24 h. TNF ? in culture supernatant was determined by ELISA following LPS stimulation. Apoptosis was determined by the TUNEL assay after treatment with either LPS or TNF ?. Results:LPS induced apoptosis in wild type AEC Ⅱ in a concentration dependent manner. LPS induced AEC Ⅱ apoptosis was accompanied by a 11 fold increase from (0.073?0.065) ng/ml in controls to( 0.94?0.14)ng/ml in 50 ?g/ml of LPS( P
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Objective To study the changes of transport protein Mrp2 excreting bilirubin across the canalicular membrane in obstructive jaundice rats. Methods The Mrp2 transcription and protein levels of hepatocytes was measured by RT PCR and immunofluorescence microscopy in obstructive jaundice rats. A correlation analysis of the transcription and protein levels with TNF ? and serum bilirubin was conducted. Results The Mrp2 transcription and protein levels were significantly lower in hepatocytes following bile duct obstruction and negatively correlated with TNF ? and bilirubin in rat serum. Conclusion Overexpression of inflammatory factor TNF ? may cause downregulation of transcription and protein levels of Mrp2 gene in obstructive jaundice rats. Bilirubin secretion across canalicular membrane can be inhibited. These may lead to or exaggerate hyperbilirubinemia.
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AIM To study the boosting effect of Maximin 2 on expression of HLA DR and HLA ABC by cultured bladder cancer cell lines and compare with TNF ??IFN ?. METHODS Cell culture and FCM were used to dectect the effects of Maximin 2,TNF and IFN on the cell growth of three types of bladder cancer cell lines and HLA DR and HLA ABC expression under the IC 50 culture concentration. RESULTS Maximin 2 had show inhibition to cells growth under the small dosage. TNF ??IFN ? also had the effect on restrain all 3 cell lines growth. The expression of HLA DR was not any change in all 3 cell lines after being stimulated by Maximin 2?TNF ? and IFN ?, Maximin 2 and TNF ? can not change HLA ABC expression, IFN ? can up regulate HLA ABC expression. CONCLUSION Maximin 2 show the dose dependent effect on cell growth suppression; The mechanism of antitumor of Maximin 2 seems not relate to the enhanced expression of HLA DR or HLA ABC, TNF ? and IFN ? also not found effect to HLA DR expression. Since IFN ? are able to increase the expression of HLA ABC expression,enhanced recognization and cytotoxity of CTL to bladder cancer cells. It is may be one of the mechanism of antitumor of IFN ?.
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Objective To investigate the relation of inflammation markers to acute coronary syndrome and the clinical significance by observing the changes of P selectin、CRP 、IL 6、TNF ?、ICAM 1 during unstable agina pectoris and acute myocardial infarction(AMI). Methods 45 patients were selected as acute coronery syndrome(ACS)group, including 25 patients with unstable agina pectoris (UAP) and 20 patients with AMI; 25 patients as stable agina pectoris (SAP)and 20 persons as control group. The blood sample of the control group are drawn from venous in the morning, the agina pectoris group are drown from venous in the morning of the second day after hospitalized, however, the AMIgroup are drown from venous at the point of 6 hour、 12 hour、24 hour、48 hour、72 hour after episode. The concentration of CRP is determined by Scatter Turbidimetry , and the concentration of P selectin、IL 6 、TNF ?、ICAM 1 by ELISA. Results (1) SAP vs control ,the levels of P selectin、CRP 、IL 6、TNF ?、ICAM 1 all raise significantly ( P 0.05) ; (4) AMI vs SAP , the levels of P selectin、CRP 、IL 6、TNF ?、ICAM 1 all raise significantly ( P
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Objective To investigate the regulatory effect of TNF ? on the expression of plasminogen activator inhibitor 2(PAI 2) and its biological significance. Methods Using immunocytochemistry(ICC), terminal deoxynucleotidyl transferase mediated end labeling (TUNEL) and ICC/TUNEL double label methods, the expression of PAI 2 in and the apoptosis of the cultured epidermal keratinocytes treated by TNF ? were detected. Results After the mouse epidermal keratinocytes were incubated with TNF ?, compared with the unshed cells, the expression of PAI 2 in shed cells and the cellular apoptosis increased obviously. The increased PAI 2 expression also appeared in the apoptotic cells, especially in larger and multi angled apoptotic cells. Conclusion In highly differentiated epidermal keratinocytes, TNF ? can enhance the expression of PAI 2 as well as apoptosis. Furthermore, according to the relation with the keratinocytes apoptosis, there are two types of PAI 2 induced by TNF ?, one of which is related to apoptosis, but the other is not.
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Objective:Study the effect of Acanthopanax gracilistylus extract(Age) on monocyte,and reveal the anti trmor mechanism.Methods:The content of TNF ?,IL 12 were examined by using biological method and ELISA,respectively.The expression of monocytic cytokine mRNA are investigated by RT PCR.The effect of Age on the phagocytosis of monocytes is studied using TdR incorporation method.Results:After cultured with Age,the production of cytokines by monocytes such as TNF ?,IL 12 are markedly increased with good dose dependent manner.After monocytes cultured with LPS in the presence or absence of Age,Age markedly enhances LPS induced TNF ? production.Treatment with Age,the production of cytokines and the expression of their mRNA are increased simultaneously.The results in vivo also show:Age can enhance the productions and the expressions of TNF ? and IL 12.Conclusion:The anti tumor active component of Age has obvious stimulative activity on monocytes, increasing the production of cytokines, enhancing the phagocytosis.These results product the scientific evidence for the study of anti tumor mechanism and clinical ultilization.
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Object To investigate the effects of ethanol sediments obtained from the tuber of Angelica sinensis (Oliv). Diels (ESA) and its litmusless component (ESA 1) on the secretion of TNF ? and IL 1 by mice peritoneal macrophages in vitro. Methods L929 cell line cytotoxicity was used for the assay of TNF ?. The proliferation of L929 cell line was used for the assay of IL 1. Results The secretion of TNF ? and IL 1 by mice peritoneal macrophages which were co cultured with ESA or ESA 1 in vitro can be significantly promoted. At the concentrations in range of 5～20 ?g/mL, there is a dose dependence in the action of ESA, while there is not the similar effect of ESA 1, even though it showed the marked effect. Conclusion ESA and ESA 1 can enhance the secreting TNF ? and IL 1 of mice peritoneal macrophages in vitro.
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Object To study the effect of emodin, the active principle in rhubarb on the secretion of TNF ?, IL 1, IL 6, and [Ca 2+ ] i by rat peritoneal macrophage under different conditions Methods Hypernomic inflammation models of isolated rat peritoneal macrophage were prepared by lipopolysaccharide excitation and the levels of [Ca 2+ ] i and the proinflammatory cytokines TNF ?, IL 1 and IL 6 secreated determined by fluorescence spectrophotometry and bioassay Results Emodin showed a dual regulatory effect on the secretion of proinflammatory cytokines and [Ca 2+ ] i Conclusion It seemed that emodin has a biphasic immuno regulatory effect
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Objective To evaluate the effect of cutting balloon angioplasty and stent implantation on serum TNF ?, IL 6 and IL 8, and investigate the relationship between the concentrations of serum TNF ?, IL 6 and IL 8 after operation and the incidences of late cardiac events after intervention Methods Eighty patients underwent PTCA with normal balloon or cutting balloon Values of serum TNF ?, IL 6 and IL 8 before and after intervention were measured using enzyme linked immuno sorbent assay Results The concentrations of serum TNF ?, IL 6 and IL 8 were increased significantly in the restenosis group, especially in the POBA group There was no obvious difference in no restenosis group Conclusion The lower restenosis rate in cutting balloon group was related to its lower injury and inflammation of blood vessels The concentrations of serum TNF ?, IL 6 and IL 8 6 hours after intervention may be the predictable sign of restenosis after intervention
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AIM To study the effects of RAPA on lymphocyte proliferation induced by mitogen and in mixed lymphocyte reaction(MLR), and the effects on T subpopulation and the content of IL 2,IFN ?,TNF ? in mice. METHODS Lymphocyte proliferation induced by mitogens and MLR were detected with MTT colometeric assay. T cell subsets were measured with fluorescence activated cell sorter (FACs). IL 2,IFN ? and TNF ? were detected by ELISA and biological assay. RESULTS RAPA inhibited murine lymphocyte proliferation induced by Con A, PHA or LPS, and also inhibited lymphocyte proliferation stimulated in MLR. RAPA had no significant effect on murine spleenic T lymphocyte subpopulation. RAPA inhibited ConA induced production of IL 2 of IFN ? by murine splenocytes, but had not inhibited LPS induced TNF ? production by murine peritoneal macrophage. CONCLUSION The immunosuppressive mechanism of RAPA is distinct from CsA.
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Objective:To investigate the changes and clinical implications of T cell subsets, immunoglobulins and tumor necrosis factor - ? (TNF- ?) of serum in schizophrenia. Methods: APAAP was used to detect T cell subsets and single immunodiffusion was used to detect immunoglobulins and ELISA was used to detect TNF- ?. Results :In 41 patients, CD3+ T cell,CD4+ T cell and CD4+CD8+T cell rates were significantly lower than normal controls(P
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Objective:To determine the region of human transmembrane tumor necrosis factor alpha (TM TNF?),essential for cytotoxic activity against mouse L929 cell.Methods:Single amino acid substituted TM TNF? mutant proteins(muteins) were produced by in vitro transcription linked translation techniques.An expression cDNA for TM TNF? was site directed mutagenized by recombinant PCR.7 single amino acid substituted TM TNF? muteins were generated and assayed for cytotoxic activity.Results:The cytotoxic activities of TM TNF? muteins,eg,TM TNF? -71/Lys was
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Objective:To investigate EB PAOA's effects on the level of cytokines produced by the mice transplanted with S 180 cells and induction actions for apoptosis of colon cancer CL 187 cells so as to probe into EB PAOA's antineoplastic active mechanism preliminarily Methods:MTT assay was used to detect EB PAOA's effects on the level of cytokines,including IL 2?IFN ??IL 1 and TNF ?,produced by the mice transplanted with S 180 cells Then, human colon cancer CL 187 cell strains were cultured in vitro and MTT assay was adopted to determine the inhibition rate to CL 187 cells Apoptotic cells were observed and confirmed with an electronmicrograph Apoptotic rate were then detected through a flow cytometer Results:The ability of the mice transplanted with S 180 cells to produce IL 2?IL 1 and TNF ? after treated with EB PAOA was strengthened EB PAOA can inhibit the proliferation of CL 187 cells Large number of apoptotic cells were found under the electronmicrograph There was an apoptotic peak appeared in DNA histogram of the flow cytometer The apoptotic rate was of time dependent.Conclusion:The antineoplastic mechanism of EB PAOA may be realized by stimulating the splenocytes of the mice transplanted with S 180 cells to secrete IL 2 and macrophages to secrete IL 1 and TNF ? to strengthen cellular immune functions and induce the apoptosis of tumor cells directly
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Objective:To study the expression of P selectin in the colonic mucosa and on surfaces of platelets in circulation, and determine its association with IL 8 and TNF ? in trinitrobenzene sulphonic acid (TNBS) induced colitis in rat, in order to investigate the effects of P selectin on inflammation and thrombosis in inflammatory bowel disease Methods:TNBS was instilled into the colon of the rats to induce colitis Animals were sacrificed at the 24th hour, the 1st and the 2nd week after production Blood and colonic tissues were obtained P selectin on the platelets were detected by radioimmunoassay IL 8 in serum was detected by ELISA method Expression of P selectin and TNF ? in colonic mucosa were assessed with immunohistochemistry Results:P selectin in mucosa was increased in different time points compared with control group The positive expression was above 50% P selectin on the platelets wasn't increased at the 24th hour, but increased at the 1st and 2nd week (P
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Objective:To study the influence and mechanism of thalidomide on bone marrow hematopoietic progenitor cells in multiple myeloma patients and in normal controls.Methods:Bone marrow GM CFU, E CFU and MK CFU from multiple myeloma patients and from normal controls were cultured in methylcellulose semisolid medium in vitro after being treated by different concentration of thalidomide. IFN ? and TNF ? concentration in cell suspension of bone marrow hematopoietic progenitor cells from multiple myeloma patients and from normal controls were measured by ELISA.Results:Thalidomide inhibited GM CFU in multiple myeloma patients and in normal controls at concentration higher than 200 and 100 ?g/ml, respectively, and inhibited MK CFU in multiple myeloma patients and in normal controls at concentration higher than 300 ?g/ml and 150 ?g/ml, respectively. However, there was no significantly influence of thalidomide on E CFU in multiple myeloma patients and in normal controls at concentration between 10 and 400 ?g/ml. Thalidomide increased IFN ? concentration in cell suspension of bone marrow hematopoietic progenitor cells from multiple myeloma patients and from normal controls at concentration greater than 100 and 200 ?g/ml, respectively. The level of thalidomide of higher than 300 ?g/ml reduced TNF ? concentration in cell suspension of bone marrow hematopoietic progenitor cells from multiple myeloma patients.Conclusion:Different concentrations of thalidomide have different influences on the hematopoietic function of bone marrow hematopoietic progenitor cells in multiple myeloma patients and in normal controls, probably mediated through increased IFN ? by thalidomide.