OBJECTIVETo study the impacts of positive antisperm antibody (AsAb) in seminal plasma on acrosomal enzyme activity, nitric oxide synthase (NOS) and superoxide dismutase (SOD) levels of spermatozoa.

METHODSSwatch from 40 infertile patients with positive AsAb in seminal plasma as experimental group, and 40 fertile men as control group. Acrosomal enzyme activity was detected by the BAEE/ADH unitive method, NOS was detected by the redoxreaction assay, and SOD level was measured xanthine oxidase method.

RESULTSCompared with control group, acrosomal enzyme activity of spermatozoa of experimental group was significantly decreased (P <0.01), NOS activity was apparently increased (P < 0.01), and SOD level in seminal plasma was markedly decreased (P<0.01).

CONCLUSIONIt may be possible that the positive AsAb in seminal plasma beget infertility through the changes of acrosomal enzyme of spermatozoa, SOD and NOS activities in seminal plasma.

Acrosin ; metabolism ; Adult ; Autoantibodies ; analysis ; Case-Control Studies ; Humans ; Infertility, Male ; enzymology ; immunology ; Male ; Nitric Oxide Synthase ; metabolism ; Semen ; enzymology ; Spermatozoa ; enzymology ; immunology ; Superoxide Dismutase ; metabolism

Excessive production of nitric oxide (NO) and proinflammatory cytokines from activated microglia play an important role in human neurodegenerative disorders. Here, we investigated whether celastrol, which has been used as a potent anti-inflammatory and anti-oxidative agent in Chinese medicine, attenuates excessive production of NO and proinflammatory cytokines such as TNF-alpha and IL-1beta in LPS-stimulated BV-2 cells, a mouse microglial cell line. We report here that the LPS-elicited excessive production of NO, TNF-alpha, and IL-1beta in BV-2 cells was largely inhibited in the presence of celastrol, and the attenuation of inducible iNOS and these cytokines resulted from the reduced expression of mRNAs of iNOS and these cytokines, respectively. The molecular mechanisms that underlie celastrol-mediated attenuation were the inhibition of LPS-induced phosphorylation of MAPK/ERK1/2 and the DNA binding activity of NF-kappaB in BV-2 cells. The results indicate that celastrol effectively attenuated NO and proinflammatory cytokine production via the inhibition of ERK1/2 phosphorylation and NF-kappaB activation in LPS-activated microglia. Thus, celastrol may be an effective therapeutic candidate for use in the treatment of neurodegenerative human brain disorders.
Animals ; Cell Line ; Cytokines/*biosynthesis/drug effects ; Gene Expression Regulation, Enzymologic/drug effects/immunology ; Inflammation/immunology ; Inflammation Mediators/immunology ; Mice ; Microglia/*drug effects/immunology ; Mitogen-Activated Protein Kinases/*physiology ; NF-kappa B/metabolism/*physiology ; Nitric Oxide/*metabolism ; Nitric Oxide Synthase Type II/biosynthesis/drug effects ; RNA, Messenger/analysis ; Signal Transduction/*drug effects/physiology ; Transcription, Genetic/drug effects/immunology ; Triterpenes/*pharmacology

OBJECTIVETo investigate the clinical value of combined determination of in vitro allergens and fractional exhaled nitric oxide (FeNO) in indentifying children at a high risk of asthma among those with recurrent wheezing.

METHODSA total of 148 children with recurrent wheezing (0.5-6 years old) were enrolled as study subjects, and 80 healthy children who underwent physical examination were enrolled as the control group. Pharmacia UniCAP immunoassay analyzer was used to measure specific immunoglobulin E (sIgE). Nano Coulomb Nitric Oxide Analyzer was used to measure FeNO. The asthma predictive index (API) was evaluated.

RESULTSThe recurrent wheezing group had a significantly higher proportion of children with positive sIgE than the control group [68.9% (102/148) vs 11.3% (9/80); P<0.05]. The recurrent wheezing group also had significantly higher levels and positive rate of FeNO than the control group (P<0.05). The overall positive rate of API in children with wheezing was 32.4%, and the API-positive children had a significantly higher FeNO value than the API-negative children (51±6 ppb vs 13±5 ppb; P<0.05). The detection rate of API was 40.2% (41/102) in positive-sIgE children and 50.1% (38/73) in FeNO-positive children, and there was no significant difference between these two groups. The children with positive sIgE and FeNO had a significantly higher detection rate of API (81.4%) than those with positive sIgE or FeNO (P<0.05).

CONCLUSIONSCombined determination of FeNO and in vitro allergens is more sensitive in detecting children at a high risk of asthma than FeNO or in vitro allergens determination alone and provides a good method for early identification, diagnosis, and intervention of asthma in children.

Allergens ; immunology ; Asthma ; diagnosis ; Breath Tests ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin E ; blood ; Infant ; Male ; Nitric Oxide ; analysis ; Recurrence ; Respiratory Sounds ; diagnosis

BACKGROUNDThe airway inflammation could be assessed by some noninvasive approaches. To investigate the value of eosinophil counts in induced sputum and fractional concentration of exhaled nitric oxide (FENO) for the regimen adjustment in patients with asthma, the correlation was analyzed between the two parameters and lung function parameter (forced expiratory volume in one second (FEV(1))).

METHODSSixty-five outpatients with mild to moderate non-exacerbation asthma from Beijing Chao-Yang Hospital were enrolled as treatment group. Combined medications of inhaled corticosteroids plus long-acting beta-2 agonist were administered for one year. Lung function parameters, eosinophil counts in induced sputum, concentration of exhaled nitric oxide and the Asthma Control Test scores were recorded, at regular intervals in the follow-up period. Twenty-one healthy volunteers were enrolled as control group and underwent examination of eosinophil counts in induced sputum, lung function and concentration of exhaled nitric oxide.

RESULTSSixty-three subjects from treatment group completed follow-up period for one year or longer. Mean FEV(1) value of the 63 subjects was (2.75 ± 0.54) L at baseline, (2.97 ± 0.56) L and (3.07 ± 0.52) L at month 3 and month 6, respectively, and maintained as (3.14 ± 0.51) L in the following six months. Mean FENO decreased from (61 ± 25) parts per billion (ppb) at baseline to (32 ± 19) ppb at month 3 (P < 0.05), and continued to decrease to (22 ± 12) ppb at month 6, the difference being significant when compared to both baseline and control group ((13 ± 8) ppb). Mean eosinophil counts decreased to (0.032 ± 0.011) × 10(6)/ml at month 3, which was significantly different from baseline ((0.093 ± 0.023)×10(6)/ml) and the control group ((0.005 ± 0.003)×10(6)/ml (both P < 0.05). The eosinophil counts in induced sputum correlated positively with concentration of FENO in the first six months (all P < 0.05). The concentration of FENO had a significant negative correlation with FEV(1) value (all P < 0.05) in any time point in the follow-up period. The Asthma Control Test scores were 18 ± 5, 19 ± 7, 23 ± 2, 24 ± 1 and 24 ± 1 at months 1, 3, 6, 9 and 12, respectively, which were significantly different from the score at baseline (14 ± 3) (P < 0.05). The most rapid clinical effect was observed at the second month after treatment.

CONCLUSIONEosinophil counts in induced sputum and FENO are sensitive parameters to detect airway inflammation and may be useful in evaluating the efficacy of treatment and adjusting medication regimens.

Adult ; Asthma ; immunology ; physiopathology ; Breath Tests ; Eosinophils ; physiology ; Female ; Humans ; Leukocyte Count ; Lung ; physiopathology ; Male ; Middle Aged ; Nitric Oxide ; analysis ; Sputum ; cytology

OBJECTIVETo investigate the effect of activated greater omental milky spots and peritoneal macrophages in mice on tumoricidal activity against gastric carcinoma SGC-7901, following intraperitoneal (i.p.) injection of INF-gamma, staphylococcin aureus or NDV-L.

METHODSThe quantitative changes of milky spots were determined by activated carbon, the number of the macrophage in milky spots was assessed by nonspecific esterase stain and the number of peritoneal macrophages was counted by trypan blue exclusion. The morphology of peritoneal macrophages was observed by scanning electron microscope, the amount of TNF-alpha and iNOS mRNA expressed by peritoneal macrophages was measured by fluorescence quantitative PCR and the cytotoxicity of peritoneal macrophages supernatant against SGC-7901 was evaluated by MTT assay.

RESULTSIt was found in the treated groups that: 1. The amount of greater omental milky spots and the macrophages in milky spots increased, 2. The number of peritoneal macrophages increased. The peritoneal macrophages were in activated status. The effect TNF-alpha and iNOS mRNA expression increased and 3. The cytotoxicity against in vitro SGC-7901 increased.

CONCLUSIONIntraperitoneal injection of IFN-gamma, staphylococcin aureus or NDV-L could activate the milky spots of the greater omentum and the macrophages in peritoneal cavity in mice, with IFN-gamma being the best. The supernatant of activated peritoneal macrophages has cytotoxicity against SGC-7901. Administration of LPS to macrophages cultured in vitro could amplify the activation and enhance the cytotoxicity of the supernatant against SGC-7901.

Animals ; Cell Line, Tumor ; Cytotoxicity, Immunologic ; Female ; Interferon-gamma ; pharmacology ; Macrophages, Peritoneal ; drug effects ; immunology ; ultrastructure ; Mice ; Nitric Oxide Synthase Type II ; genetics ; Omentum ; drug effects ; immunology ; ultrastructure ; RNA, Messenger ; analysis ; Stomach Neoplasms ; immunology ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo establish an HPLC-ELSD fingerprint of the whole herbs of Morina nepalensis and perform the correlation analysis of chemical components of the herb and nitric oxide (NO) production inhibition.

METHODHPLC-ELSD assay was performed to evaluate 10 batches of M. nepalensis herbs. The chromatographic conditions were as following: Eclipse XDB C18 column (4.6 mm x 150 mm, 5 microm), water (A) and acetonitrile (B) as a gradient mobile phases, flow rate 1.0 mL x min(-1), and column temperature at 35 degress C. Evaporative light-detection conditions: atomization temperature at 104 degrees C, the flow rate of N2 2.8 L x min(-1) and 10 microL sample injection. Chromatographic fingerprint was developed, and the inhibition activity of production of NO in lipopolysaccharide-induced macrophages was also analyzed. The similarity and correlation analysis between the HPLC-ELSD fingerprints and NO production inhibition were carried out by PLS method.

RESULTThe common mode for M. nepalensis herb fingerprint was established, including 15 common characteristic peaks. Among them, 7 peaks were positively correlated with the NO production inhibition. According to the assessment on the similarity of 10 batches of samples, a similarity of over 0.90 were shown in HPLC-ELSD fingerprint and all samples were separated into two groups.

CONCLUSIONThis method can be used to assess the quality of M. nepalensis, which provides a reliable method for scientific assessment and quality control.

Animals ; Anti-Inflammatory Agents ; analysis ; pharmacology ; Caprifoliaceae ; chemistry ; Cell Line ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Macrophages ; drug effects ; immunology ; Medicine, Tibetan Traditional ; Mice ; Nitric Oxide ; immunology

We investigated the immunostimulatory effects of a novel beta-glucan purified from Paenibacillus (P.) polymyxa JB115 on bone marrow-derived dendritic cells (DCs), a type of potent antigen-presenting cells. beta-glucan isolated from P. polymyxa JB115 enhanced the viability and induced the maturation of DCs. beta-glucan markedly increased the cytokine production of DCs and surface expression of DC markers. In addition, DCs treated with beta-glucan showed a higher capacity to stimulate allogeneic spleen cell proliferation compared to those treated with medium alone. These results demonstrate the effect of beta-glucan on DC maturation and may increase the use of beta-glucan.
Animals ; Bone Marrow Cells/cytology/*drug effects/*immunology ; Cell Survival/drug effects/*immunology ; Dendritic Cells/cytology/*drug effects/*immunology ; Flow Cytometry ; Immunophenotyping/methods ; Interleukin-12/analysis/immunology ; Mice ; Mice, Inbred BALB C ; Nitric Oxide/analysis/immunology ; Paenibacillus/*chemistry ; Tumor Necrosis Factor-alpha/analysis/immunology ; beta-Glucans/isolation & purification/*pharmacology

Exhaled nitric oxide (eNO) has been proposed as a noninvasive marker of airway inflammation in asthma. In asthmatic patients, exhaled NO levels have been shown to relate with other markers of eosinophilic recruitment, which are detected in blood, sputum, bronchoalveolar lavage fluid and bronchial biopsy samples. The purpose of this study was to assess the possible relationship between eNO and allergic inflammation or sensitization in childhood asthma and allergic rhinitis. Subjects consisted of 118 asthmatic children, 79 patients with allergic rhinitis, and 74 controls. Their age ranged from 6 to 15 yr old. eNO level, peripheral blood eosinophil count, eosinophil cationic protein (ECP), serum total IgE level and specific IgE levels were measured. Methacholine challenge test and allergic skin prick test for common allergens were performed in all subjects. Atopic group (n = 206, 44.48 +/- 30.45 ppb) had higher eNO values than non-atopic group (n = 65, 20.54 +/- 16.57 ppb, P < 0.001). eNO level was significantly higher in patients with asthma (42.84 +/- 31.92 ppb) and in those with allergic rhinitis (43.59 +/- 29.84 ppb) than in healthy controls (27.01 +/- 21.34 ppb, P < 0.001) but there was no difference between asthma and allergic rhinitis group. eNO also had significant positive correlations with Dermatophagoides pteronyssinus IgE level (r = 0.348, P < 0.001), Dermatophagoides farinae IgE level (r = 0.376, P < 0.001), and the number of positive allergens in skin prick test (r = 0.329, P = 0.001). eNO had significant positive correlations with peripheral blood eosinophil count (r = 0.356, P < 0.001), serum total IgE level (r = 0.221, P < 0.001), and ECP (r = 0.436, P < 0.001). This study reveals that eNO level is associated with allergic inflammation and the degree of allergic sensitization.
Adolescent ; Allergens/immunology ; Animals ; Asthma/*immunology ; *Breath Tests ; Bronchial Provocation Tests ; Child ; Dermatophagoides pteronyssinus/immunology ; Eosinophil Cationic Protein/analysis/blood/immunology ; Eosinophils ; Exhalation ; Female ; Humans ; Hypersensitivity, Immediate/*immunology ; Immunoglobulin E/blood ; Leukocyte Count ; Male ; Nitric Oxide/*analysis ; Rhinitis, Allergic, Seasonal/*immunology

Adolescent ; CD3 Complex ; blood ; CD4 Antigens ; blood ; CD8 Antigens ; blood ; Child ; Fetal Blood ; immunology ; Humans ; Infant, Newborn ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Male ; Nitric Oxide ; blood ; T-Lymphocytes ; immunology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVENuclear factor-kappa B (NF-kappaB) is a critical transcription factor governing the expression of many cytokines that are involved in the pathogenesis of inflammatory diseases, such as asthma and rheumatoid arthritis. Melatonin (MT), a relatively safe and potent antioxidant which has shown efficacy in several chronic inflammatory models, may inhibit the expression of NF-kappaB and therefore might have a therapeutic use in asthma. This study aimed at observing the effect of MT on the expression of NF-kappaB and airway inflammation in a rat model of bronchial asthma.

METHODSTwenty-four male Sprague-Dawley (SD) rats weighing 120 g to 170 g were randomly divided into three experimental groups (8 in each): (1) Asthmatic group: Rats were immunized on day 1 by intraperitoneal injection of 100 mg ovalbumin (OVA) in 1 ml of saline with 100 mg of alu minum hydroxide. From day 15 the animals were challenged with aerosolized OVA (1% in saline) for 20 minutes per day for 7 consecutive days. (2) MT group: OVA-sensitized rats were injected intraperitoneally with 10 mg/kg MT 30 minutes before each OVA challenge. (3) CONTROL GROUP: OVA for inhalation and MT for intraperitoneal injection was replaced with normal saline (NS). Airway responsiveness to aerosolized acetylcholine of 24 rats was detected six hours after the last challenge. Then the rats were lavaged and total and differentiated leukocytes counts in bronchoalveolar lavage fluid (BALF) were performed after staining with Wright-Giemsa staining. At the same time, levels of nitric oxide (NO) in BALF, inducible nitric oxide synthesis (iNOS) and constitute nitric oxide synthesis (cNOS) in the lung tissues were assessed with the use of nitrate reductase and chemical colorimetry, respectively. The expression of NF-kappaB in the lung tissues was observed by means of immunohistochemical staining.

RESULTS(1) After OVA challenge, there was a significant decrease in airway responsiveness, lymphocytes and eosinophils in BALF in MT group compared with asthmatic group (P < 0.01 respectively); (2) There was a significant decrease in amounts of NO(2)(-)/NO(3)(-) in the BALF and levels of iNOS in the lung tissues in MT group comparing with asthmatic group (P < 0.01 respectively); and the levels of iNOS in the lung tissues was correlated positively with NO(2)(-)/NO(3)(-) in the BALF (P < 0.01), but there were no significant differences in activity of cNOS in any of the groups analyzed. (3) There was a significant increase in expression of NF-kappaB in lung tissues in asthmatic group compared with the other groups (P < 0.01), and so was in MT group compared with control group (P < 0.05).

CONCLUSIONSMT could partially inhibit the expression of NF-kappaB and down-regulate the activity of iNOS in lung tissue, decrease the production of NO in BALF. These data suggest that the inhibitory effect of MT probably play a role in decreasing airway hyperresponsiveness and airway inflammation of asthmatic rats model.

Animals ; Antioxidants ; pharmacology ; Asthma ; drug therapy ; metabolism ; Bronchoalveolar Lavage Fluid ; cytology ; immunology ; Disease Models, Animal ; Lung ; chemistry ; pathology ; Male ; Melatonin ; pharmacology ; NF-kappa B ; analysis ; Nitric Oxide ; analysis ; Rats ; Rats, Sprague-Dawley