OBJECTIVETo determine the clinical significance of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha( TNF-alpha) in expressed prostatic secretions(EPS) for chronic prostatitis.

METHODSProstatic secretions IL-1beta and TNF-alpha were evaluated for 34 patients with chronic prostatitis, 10 with asymptomatic inflammatory prostatitis, 12 with benign prostatic hyperplasia (BPH) and 8 health controls by enzyme-linked immunosorbent assay (ELISA).

RESULTSIL-1beta and TNF-alpha levels in EPS in the patients of chronic prostatitis with WBC > or = 10/HP and asymptomatic inflammatory prostatitis were obviously higher than those of chronic prostatitis with WBC < 10/HP, BPH and health controls, (P < 0.05 and P < 0.02). There was a correlation between IL-1beta and TNF-alpha (P < 0.003) but none between WBC and IL-1beta or TNF-alpha.

CONCLUSIONCytokines are frequently elevated in EPS in men of chronic prostatitis with high WBC and asymptomatic inflammatory prostatitis, which provides a novel means different from traditional methods based on WBC for the identification of men with chronic prostatitis.

Aged ; Chronic Disease ; Humans ; Interleukin-1 ; analysis ; Male ; Middle Aged ; Prostate ; chemistry ; secretion ; Prostatitis ; immunology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVEBy using RAW 264.7 macrophage cell line, we studied the dose-effect relationship of endotoxin induced RAW 264.7 cells to release TNF-alpha, and then detected the content of endotoxin in 8 kinds of injections, so that we can investigate the feasibility and the interference factors of the novel test.

METHODBy using endotoxin of different concentrations to induce RAW 264. 7 cells to release TNF-a, we drew the curve of dose-effect relationship between endotoxin and generated TNF-alpha. Then we detected the content of TNF-alpha in yuxingcao, shuanghuanglian, qingkailing, gegensu, xiangdan, qianrongmei and jiangxianmei injections and shuanghuanglian powder injection, and calculated their content of endotoxin.

RESULTThe endotoxin could induce the cells to release TNF-alpha in a good dose-dependent manner, even at a very low concentration. In the range of maximum available dilution multiple, the content of endotoxin in the rest 7 kinds of injections was less than 1.0 EU x mL(-1) except qingkailing injection of two batch.

CONCLUSIONCytokine revulsion has the advantage of wide detection range, high sensitivity, simple operation, and the detected endotoxin is of bioactivity. This method provides another technical mean for pyrogen test of injections.

Animals ; Biological Assay ; methods ; Cell Line ; Drug Contamination ; Drugs, Chinese Herbal ; analysis ; Endotoxins ; adverse effects ; analysis ; Macrophages ; drug effects ; immunology ; Mice ; Tumor Necrosis Factor-alpha ; analysis ; immunology

OBJECTIVETo study the levels and roles of cytokines TNF-&alpha;, IL-6 and IL-10 in bronchoalveolar lavage fluid (BALF) in children with Mycoplasma pneumoniae pneumonia (MPP).

METHODSThe levels of TNF-&alpha;, IL-6 and IL-10 in BALF were measured using ELISA in children with MPP at acute stage (n=45) and at remission stage (n=30). Twenty children without lung lesions severed as the control group.

RESULTSThe TNF-&alpha;, IL-6 and IL-10 levels in BALF were higher in children with MPP at acute stage than those in the control group (P<0.05). The levels of TNF-&alpha; and IL-6 in BALF at remission stage were reduced to the levels similar to the control group and were significantly lower than those at the acute stage in children with MPP. However, the levels of IL-10 in BALF remained at higher levels at remission stage in children with MPP.

CONCLUSIONSThe levels of TNF-&alpha;, IL-6 and IL-10 in BALF increase in children with MPP at acute stage, suggesting that the cytokines may be involved in the pathogenesis of MPP.

Adolescent ; Bronchoalveolar Lavage Fluid ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Interleukin-10 ; analysis ; Interleukin-6 ; analysis ; Male ; Pneumonia, Mycoplasma ; etiology ; immunology ; Tumor Necrosis Factor-alpha ; analysis

The development of the systematic evolution of ligands by exponential enrichment (SELEX) process has made it possible to isolate oligonucleotide sequences with the capacity of recognizing virtually any class of target molecules with high affinity and specificity. These oligonucleotide sequences, referred to as "aptamers", are useful as a class of molecules that rival antibodies in diagnostic applications. Aptamers are different from antibodies, yet they mimic properties of antibodies in a variety of diagnostic formats. To meet the shortcomings of antibodies, aptamers have the following advantages. Aptamer does not depend on animals, cells, or even in vivo conditions and produced by chemical synthesis with extreme accuracy and reproducibility. Once denatured, functional aptamers could be regenerated easily within minutes. They are stable to long-term storage and can be transported at ambient temperature. We describe here an enzyme -linked oligonucleotide assay that use a SELEX-derived RNA aptamer to detect hTNFalpha. In order to protect from nuclease attack, the RNA aptamer was modified by replacement of 2'-NH2 for 2'-OH at all ribo-purines. In a sandwich micro-plate assay, hTNFalpha monoclonal antibody was coated on the surface of the plate, biotin-labeled RNA aptamer was used as a detect molecle. HTNFalpha was diluted by pooled human serum as standard, and streptavidin-horseradish peroxidase-substrate system was added for detection. Accuracy, precision, sensitivity, specificity of ELONA method were analyzed. The levels of hTNF-alpha in normal human serum samples were assayed by the ELONA and the ELISA processes. The resultes demonstrate that a sandwich assay using a SELEX-derived RNA aptamer has parameters for accuracy, precision, sensitivity, specificity well within the limits expected of a typical enzyme-linked assay. There is no significant difference between the results of ELONA and ELISA. The minimum detection level was 100 pg/mL. This method will be useful for detection of almost all the cytokines and other protein molecules.
Enzyme-Linked Immunosorbent Assay ; Humans ; SELEX Aptamer Technique ; methods ; Sensitivity and Specificity ; Tumor Necrosis Factor-alpha ; analysis ; immunology

Adult ; Female ; Hepatitis B ; immunology ; Humans ; Interferon-gamma ; blood ; Interleukin-12 ; blood ; Male ; Middle Aged ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo observe the effect of recombinant human erythropoietin (rHuEPO) on immune function of premature rats.

METHODSRHuEPO of 250 IU/(kg.t) or 500 IU/(kg.t) was administered to premature rats every other day for nineteen days. The control premature rats were received normal saline. The changes of hemoglobin (Hb), serum erythropoietin (EPO), red blood cell (RBC) immune function, T lymphocyte proliferative responsiveness, and production of tumor necrosis factor alpha (TNF-alpha) were observed.

RESULTSPremature rats showed lower levels on Hb, RBC immune function, T cell responsiveness and production of TNF-alpha compared with mature rats at birth. The postnatal declines of Hb and RBC immune function were lessened in the treated groups of premature rats, the higher dosage group of 500 IU/(kg.t) was more significant than the lower dosage group of 250 IU/(kg.t). When experiments were over, Hb of control premature rats was (7.72 +/- 0.89) g/dl, Hb of premature rats received 500 IU/(kg.t) was (10.08 +/- 0.90) g/dl (P < 0.01). C3b-R% of control premature rats was (11.00 +/- 0.95)%, C3b-R% of premature rats received 500 IU/(kg.t) was (17.75 +/- 1.04)% (P < 0.01). IC-R% in control premature rats was (12.83 +/- 1.33)%, IC-R% of premature rats received 500 IU/(kg.t) was (10.50 +/- 1.67)% (P < 0.01). The postnatal rise of T cell responsiveness and the production of TNF-alpha in premature rats increased in the treated groups, which was more significant in the higher dosage group of 500 IU/(kg.t) than in the lower dosage group of 250 IU/(kg.t). The OD index of control premature rats was 0.159 +/- 0.014, the OD index of premature rats received 500 IU/(kg.t) was 0.354 +/- 0.050 (P < 0.01). TNF-alpha in control premature rats was (0.270 +/- 0.014) ng/ml, TNF-alpha of premature rats received 500 IU/(kg.t) was (0.415 +/- 0.010) ng/ml (P < 0.01).

CONCLUSIONS(1) Premature rats had lower RBC immune function and T cell responsiveness and underproduction of TNF-alpha at birth. (2) Premature rats had an improvement with the RBC immune function after rHuEPO administration. (3) Premature rats had improvements with T cell responsiveness and TNF-alpha after rHuEPO administration, and there was a positive correction between the RBC immune function and T cell responsiveness with the production of TNF-alpha.

Animals ; Erythrocyte Count ; Erythrocytes ; drug effects ; immunology ; Erythropoietin ; blood ; pharmacology ; Female ; Hemoglobins ; analysis ; Pregnancy ; Rats ; Recombinant Proteins ; T-Lymphocytes ; drug effects ; immunology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo investigate the regulating effect of Ginkgo biloba extract 50 (GBE50) on pre-inflammatory factors interleukin-1 beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and anti-inflammatory factors interleukin-4 (IL-4), interleukin-10 (IL-10) of hippocampus in senile rats, in order to explore the protective mechanism of GBE50 on central nervous system of senile animals.

METHODSD rats were randomly divided into four groups: the normal group, the model group, the GBE50 group and the EGB761 group. Rats were intraperitoneally injected with 100 mg x kg(-1) D-galactose every day for 42 days to establish the senile rat model. At the 21st day, the GBE50 group and the EGB761 group were orally administered with 60 mg x kg(-1) for 21 days. IL-1beta mRNA and TNF-alpha mRNA expressions were detected by real-time fluorescence quantitative PCR assay, IL-1beta and TNF-alpha protein expressions were detected by immunohistochemistry, IL-4 and IL-10 protein contents were detected by ELISA.

RESULTD-galactose caused imbalance between pre-inflammatory factors and anti-inflammatory factors of hippocampus in senile rats, GBE50 and EGB761 reduced IL-1beta mRNA expression (P < 0.05) and TNF-alpha and IL-1beta protein level (P < 0.01) and up-regulated IL-10 protein content (P < 0.01, P < 0.05).

CONCLUSIONThe mechanism of GBE50 in protecting central nervous system is probably related to its effect in mitigating inflammatory of central nervous system.

Animals ; Ginkgo biloba ; Hippocampus ; drug effects ; immunology ; Interleukin-1beta ; analysis ; Male ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; analysis

To explore the role of immune regulating cytokines in pathogenesis of the idiopathic thrombocytopenic purpura (ITP) and its clinical significance, the levels of IL-18, TNF-alpha and Sc5b-9 in plasma of 32 ITP patients and 18 normal individuals were detected using ELISA methods. The results showed that IL-18, TNF-alpha and sC5b-9 levels in plasma of ITP patients were higher than that in normal individuals. The level of IL-18 was positively correlated with the levels of TNF-alpha and sC5b-9. In conclusion, The rising levels of the IL-18, TNF-alpha and sC5b-9 were correlated with disorder of Th1/Th2 subsets, and may contribute to the immune dysfunction in ITP patients. The dynamic observation of these cytokines may be useful in directing the clinical treatment for ITP patients.
Adolescent ; Adult ; Child ; Child, Preschool ; Complement Membrane Attack Complex ; analysis ; Female ; Humans ; Interleukin-18 ; blood ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo investigate the proinflammatory cytocine factors in patients with coronary artery disease by different treatments during perioperation.

METHODSTNF-alpha, IL-6, c-reactive protein (CRP) were measured in 37 patients with coronary artery disease (CAD) and 10 patients with mitral valve replacement (MVR, control group) before operation and aortic clamping or before coronary artery bypass grafting (CABG) in the patients with cardiopulmonary bypass, or before transmyocardial laser reperfusion (TMLR), when aortic declamping and 3, 6, 24 hours after operation.

RESULTSThe level of proinflammatory factors increased more significantly after operation than before operation. TNF-alpha increased more significantly in the patients with CPB than in those without CPB after operation [(4.10 +/- 0.71) pg/ml vs. (1.34 +/- 0.29) pg/ml, P < 0.05)]. IL-6 was not the different among all groups after operation. CRP was higher in the CAD patients with CPB than in the control group after operation [(12.89 +/- 0.29) pg/ml vs. (12.00 +/- 0.31) pg/ml, P < 0.05].

CONCLUSIONInflammatory response can be seen after operation in all CAD patients. In patients undergoing CABG without CPB or undergoing TMLR, the changes in flammatory response are milder than those in patients with CPB. Thus patients should undergo CABG without CPB if they are indicated.

Aged ; C-Reactive Protein ; analysis ; Cardiopulmonary Bypass ; Coronary Artery Bypass ; Coronary Disease ; immunology ; surgery ; Female ; Humans ; Interleukin-6 ; blood ; Male ; Middle Aged ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo understand the effects of the IL-1, TNF-alpha cytokines on the pathogenesis of periapical lesions by investigating its gene expression in rat.

METHODSThe model was established by surgically exposing mandibular molar teeth and left open to permit infection from the oral environment. The SD rats were killed at 1, 2, 3, 4 weeks and mandibular molar teeth X-ray were taken. In situ hybridization was used to test IL-1alpha, IL-1beta, TNF-alpha mRNA expression in periapical area and the kinds of positive cells were identified. Using image analysis system analyzed gene expression semi-quantified.

RESULTSIL-1alpha, IL-1beta and TNF-alpha mRNA were all expressed beginning at 1 week, peaked at 3 weeks, and declined somewhat at 4 weeks, but IL-1beta mRNA was expressed at much lower levels with the same kinetics (P<0.01). Most of the staining occurred in areas that had heavy inflammatory infiltrate, fibroblasts, or endothelial cells. There was a statistically significant correlations between the area of periapical lesion and the number of positively stained cells for IL-1alpha and for TNF-alpha (IL-1alpha: r=0.875, P<0.001; TNF-alpha: r=0.858, P<0.001), and between the number of positive cells for IL-1alpha and that of for TNF-alpha (r=0.969, P<0.001).

CONCLUSIONSIL-1alpha and TNF-alpha genes are highly expressed in developing periapical lesions in rat, which supports the hypothesis that these two cytokines play a key role in pulpal and periapical pathogenesis, including the concomitant bone destruction.

Animals ; In Situ Hybridization ; Interleukin-1 ; genetics ; physiology ; Periapical Periodontitis ; etiology ; immunology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; physiology