Plasma Inflammatory Cytokine IL-4, IL-8, IL-10, and TNF-α Levels Correlate with Pulmonary Function in Patients with Asthma-Chronic Obstructive Pulmonary Disease (COPD) Overlap Syndrome.

BACKGROUND The aim of this study was to investigate the plasma inflammatory cytokine levels and their correlations with pulmonary function in patients with asthma-chronic obstructive pulmonary disease overlap syndrome (ACOS). MATERIAL AND METHODS Between January 2013 and December 2014, a total of 96 patients with asthma, acute exacerbation of chronic obstructive pulmonary disease (AECOPD), or ACOS were enrolled, and 35 healthy people were included as a control group. Fasting plasma interleukin (IL)-4, IL-8, IL-10, and tumor necrosis factor alpha (TNF-α) levels were detected using enzyme-linked immunosorbent assay (ELISA). Correlations between the plasma inflammatory cytokine levels and forced expiratory volume in 1 second (FEV1), FEV1/predicted value ratio (FEV1%pred), and FEV1/forced vital capacity (FVC) were analyzed. RESULTS IL-4 and IL-8 levels showed statistically significant differences among the 3 groups of patients (both P<0.001); IL-4 level was significantly lower, while IL-8 level was significantly higher in the AECOPD group and ACOS group than those in the asthma group (all P<0.05). IL-10 level and TNF-α level were significantly different among the 3 patient groups (both P<0.001). IL-10 level was significantly different between each of the 2 groups (all P<0.001). TNF-α level in the asthma group was higher than in the AECOPD group and ACOS group (both P<0.001). IL-4 and IL-10 were positively and IL-8 and TNF-α were negatively related with FEV1, FEV1%pred, and FEV1/FVC. CONCLUSIONS Plasma levels of inflammatory cytokines IL-4, IL-8, IL-10, and TNF-α are related with severity of airway diseases and could be potential markers for the evaluation of asthma, COPD, and ACOS.

Copper amine oxidase-catalysed hydrogen peroxide involves production of nitric oxide in darkness-induced stomatal closure in broad bean.

Hydrogen peroxide is an important intermediate in darkness-induced stomatal closure. In the present work, we provide evidence that copper amine oxidase (CuAO) was involved in H2O2 production in darkness-induced stomatal closure in Vicia faba L. Darkness activated CuAO in intercellular washing fluid from leaves. Aminoguanidine (AG) and 2-bromoethylamine (BEA), which were both irreversible inhibitors of CuAO, significantly suppressed darkness-induced stomatal closure and H2O2 generation. The effects of AG and BEA were reversed only by H2O2 but not by other products of CuAO. These results indicate that CuAO participates in darkness-induced stomatal closure through its reaction product, H2O2. Furthermore, darkness-induced nitric oxide (NO) production and cytosolic alkalinisation were obviously inhibited by AG and BEA, and only H2O2, among the products of CuAO, could reverse the effects, implying that the CuAO-catalysed product H2O2 is required for NO production and cytosolic alkalinisation to a large extent in darkness-induced stomatal closure. In addition, butyric acid blocked but methylamine enhanced the ability of H2O2 to reverse the effect of BEA on NO production, suggesting that cytosolic alkalinisation is involved in CuAO-mediated NO generation in darkness-induced stomatal closure.

Inhibition of ABA-induced stomatal closure by fusicoccin is associated with cytosolic acidification-mediated hydrogen peroxide removal.

BACKGROUND: Fusicoccin (FC), a fungal phytotoxin produced by Fusicoccum amygdale, causes the inhibition of ABA-induced stomatal closure. The mechanism of inhibition is remaining unclear. We analyzed the role of hydrogen peroxide (H2O2) and relationship between H2O2 removal and cytosolic pH changes during inhibition of ABA-induced stomatal closure by FC. RESULTS: According to the results, ABA treatment induced H2O2 production and stomatal closure, but FC inhibited the effects of ABA on these two parameters. Treatment with catalase (CAT) and NADPH oxidase inhibitor diphenylene iodonium (DPI) mimicked the effect of FC. These data suggest that inhibition of ABA effect by FC is related to the decrease of H2O2 levels in guard cells. Furthermore, similar to CAT, FC not only suppressed stomatal closure and H2O2 levels in guard cells treated with exogenous H2O2, but also reopened the stomata which had been closed by ABA and reduced the level of H2O2 that had been produced by ABA, indicating that FC causes H2O2 removal in guard cells. The butyric acid treatment simulated the effects of FC on the stomatal aperture and H2O2 levels in guard cells treated with exogenous H2O2 and had been closed by ABA, and both FC and butyric acid reduced cytosolic pH in guard cells of stomata treated with H2O2 and had been closed by ABA, which demonstrate that cytosolic acidification mediates FC-induced H2O2 removal. CONCLUSION: These results suggest that FC causes cytosolic acidification in guard cells, then induces H2O2 removal and reduces H2O2 levels in guard cells, finally inhibits stomatal closure induced by ABA.

Actin microfilaments and vacuoles are downstream targets of H2O2 signalling pathways in hyperosmotic stress-induced stomatal closure.

Changes in osmotic pressure can induce stomatal closure to reduce transpirational water loss from plants. In the present work, we investigated the mechanism underlying the perception and transduction of extracellular changes in osmotic pressure in Vicia faba L. guard cells. Using an epidermal strip bioassay and laser-scanning confocal microscopy, we provide evidence that hyperosmotic stress treatment led to stomatal closure and the rapid promotion of hydrogen peroxide (H2O2) production in V. faba guard cells. The effects were largely reduced by H2O2 scavengers ASA, CAT, NADPH oxidase inhibitor DPI and cell wall peroxidase inhibitor SHAM. These results indicate that hyperosmotic stress induces stomatal closure by promoting H2O2 production. Cytochalasin B (CB), latrunculin B (Lat B) and jasplakinolide (JK) inhibited stomatal closure induced by hyperosmotic stress but didn't prevent the increase of endogenous H2O2 levels, suggesting that microfilaments reorganisation participates in stomatal closure induced by hyperosmotic stress, and may act downstream of H2O2 signalling processes. In addition, we observed splitting of big vacuoles into many small vacuoles in response to hyperosmotic stress and H2O2 treatment, and CB inhibited these changes of vacuoles; stomatal closure was also inhibited. Taken together these results indicate that the stomatal closure in response to hyperosmotic stress may initiate H2O2 generation, and that reorganisation of microfilaments and the changing of vacuoles occurs downstream of H2O2 signalling processes.

Inhibition of dark-induced stomatal closure by fusicoccin involves a removal of hydrogen peroxide in guard cells of Vicia faba.

Fusicoccin (FC) treatment prevents dark-induced stomatal closure, the mechanism of which is still obscure. By using pharmacological approaches and laser-scanning confocal microscopy, the relationship between FC inhibition of dark-induced stomatal closure and the hydrogen peroxide (H2O2) levels in guard cells in broad bean was studied. Like ascorbic acid (ASA), a scavenger of H2O2 and diphenylene iodonium (DPI), an inhibitor of H2O2-generating enzyme NADPH oxidase, FC was found to inhibit stomatal closure and reduce H2O2 levels in guard cells in darkness, indicating that FC-caused inhibition of dark-induced stomatal closure is related to the reduction of H2O2 levels in guard cells. Furthermore, like ASA, FC not only suppressed H2O2-induced stomatal closure and H2O2 levels in guard cells treated with H2O2 in light, but also reopened the stomata which had been closed by darkness and reduced the level of H2O2 that had been generated by darkness, showing that FC causes H2O2 removal in guard cells. The butyric acid treatment simulated the effects of FC on the stomata treated with H2O2 and had been closed by dark, and on H2O2 levels in guard cells of stomata treated with H2O2 and had been closed by dark, and both FC and butyric acid reduced cytosol pH in guard cells of stomata treated with H2O2 and had been closed by dark, which demonstrates that cytosolic acidification mediates FC-induced H2O2 removal. Taken together, our results provide evidence that FC causes cytosolic acidification, consequently induces H2O2 removal, and finally prevents dark-induced stomatal closure.

Nitric oxide, actin reorganization and vacuoles change are involved in PEG 6000-induced stomatal closure in Vicia faba.

Water deficit and the resulting osmotic stress affect stomatal movement. There are two types of signals, hydraulic and chemical signals, involving in the regulation of stomatal behavior responses to osmotic stress. Compared with the chemical signals, little has been known about the hydraulic signals and the corresponding signal transduction network and regulatory mechanisms. Here, using an epidermal-strip bioassay and laser-scanning confocal microscopy, we provide evidence that nitric oxide (NO) generation in Vicia faba guard cells can be induced by hydraulic signals. We used polyethylene glycol (PEG) 600 to simulate hypertonic conditions. This hydraulic signal led to stomatal closure and rapid promotion of NO production in guard cells. The effects were decreased by NO scavenger 2-(4-carboxyphenyl)-4,4,5, 5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO) and NO synthase (Enzyme Commission 1.14.13.39) inhibitor N(G)-nitro-L-Arg-methyl ester (L-NAME). These results indicate that PEG 6000 induces stomatal closure by promoting NO production. Cytochalasin B (CB) inhibited stomatal closure induced by PEG 6000 but did not prevent the increase of endogenous NO levels, indicating that microfilaments polymerization participate in stomatal closure induced by PEG 6000, and may act downstream of NO signaling. In addition, big vacuoles split into many small vacuoles were observed in response to PEG 6000 and sodium nitroprusside (SNP) treatment, and CB inhibited these changes of vacuoles, the stomatal closure was also been inhibited. Collectively, these results suggest that the stomatal closure induced by PEG 6000 may be intimately associated with NO levels, reorganization of actin filaments and the changes of vacuoles, showing a crude outline of guard-cells signaling process in response to hydraulic signals.

[Clinical study on treatment of childhood Henoch-Schonlein purpura nephritis with colquhounia root tablet].

OBJECTIVE: To study the effect of Colquhounia root tablet (CRT) in treating childhood Henoch-Schonlein purpura nephritis (HSPN) and compared with tripterygium wilfordii multiglycoside tablet (TWMGT). METHODS: Eighty-two children with HSPN were divided into the CRT group and the control group. To patients of nephritic type, treatment with CRT and TWMGT was applied separately, but to those of nephrotic type, the treatment was given together with prednisone. Therapeutic effect and changes of related indexes, including urinary protein, retinal-binding protein (RBP), N-acetyl-beta-glucosaminidase (NAG), were observed after patients received treatment for 6 months. RESULTS: The complete remission (CR) rate and partial remission (PR) rate in patients of nephritic type in the treated group was 58.8% and 41.2% respectively, while those in the control group was 20.8% and 66.7% respectively, and the ineffective rate being 12.5%. Comparison of the therapeutic effect between the two groups showed significant difference (P < 0.05). Although the CR rate in patients of nephrotic type in the treated group was higher than that in the control group, the difference was insignificant (P > 0.05). The urinary levels of protein, RBP and NAG lowered in both groups, but level of urinary protein in the treated group of nephritic type after 6 months of treatment was significantly lower than that in the control group (P < 0.05). CONCLUSION: CRT shows obvious effect in improving childhood HSPN of nephritic type, nephrotic type and attenuating the tubulointerstitial lesions, compared with TWMGT, CRT is more effective in lowering proteinuria and better in total effective rate.

[A clinico-pathological study comparing Henoch-Schonlein purpura nephritis with IgA nephropathy in children].

OBJECTIVE: Henoch-Schonlein purpura nephritis (HSPN) and IgA nephropathy are very similar in immunopathological changes, and therefore some nephrologists considered that they are substantially one disease entity caused by IgA immune abnormalities, and IgA nephropathy is, in fact, a kind of HSPN without rashes. The present study aimed to characterize their relationship through clinico-pathological comparison between IgA nephropathy and HSPN. METHODS: Thirty-one children with IgA nephropathy aged from 3 to 15 years and 120 children with HSPN aged between 4 to 15 years were enrolled in this study. Their clinical manifestations, blood biochemical test, serum immunology and follow-up data were collected and analyzed. Renal pathological findings in light microscopy, immunofluorescence and electron microscopy were analyzed and also compared between 31 children with IgA nephropathy and 32 children with HSPN. RESULTS: The age of onset was over 12 years in 25.8% children with IgA nephropathy, but only in 10% with HSPN, and the difference was significant (P < 0.05). The clinical patterns of IgA nephropathy and HSPN were similar, but extra-renal manifestations were seen more often in HSPN, all of them had skin purpura, 59% had gastrointestinal symptoms and 47% suffered from arthralgia. While the abdominal pain occurred only in 3.2% children with IgA nephropathy. The renal pathological investigation showed global sclerosis in 35.5% of IgA nephropathy and 3.1% of HSPN, mesangial sclerosis in 41.9% of IgA nephropathy and 6.3% of HSPN, but endothelial proliferation in 65.6% of HSPN and 29% of IgA nephropathy. The differences were extremely significant (P < 0.01). Thin basement membrane nephropathy were only found in 6.5% children with IgA nephropathy, but in none with HSPN. The electronic dense deposits in HSPN were sparse, loose and widely spread in glomerular mesangium, subendothelial area and even intra basement membrane. While the deposits were dense, lumpy and mostly limited in mesangium and paramesangium in IgA nephropathy. IgG was found in glomerular immune deposits in 71.9% of HSPN, but only 19.4% of IgA nephropathy. No IgG deposit was observed in 81.6% of IgA nephropathy, among them most showed IgA and IgM and/or C(3) deposit. Predominant IgG deposits were found in 12.5% of HSPN with relatively weak IgA deposit, moreover 6.3% of HSPN showed linear IgG deposits in glomerular capillary wall, which couldn't be found in IgA nephropathy. The follow-up data of average 20 months showed complete remission in 72.5% of HSPN and 19.4% in IgA nephropathy after 34 months follow-up. Moreover, 64.5% of IgA nephropathy had consistent hematuria and proteinuria, 16.1% had active nephritides, the difference was significant (P < 0.05). CONCLUSION: Significant clinico-pathological differences were found between HSPN and IgA nephropathy, which does not support the one disease entity hypothesis. HSPN and IgA nephropathy are probably two diseases with similar immune abnormalities.