Identification of cough-variant asthma phenotypes based on clinical and pathophysiologic data.

BACKGROUND: Cough-variant asthma (CVA) may respond differently to antiasthmatic treatment. There are limited data on the heterogeneity of CVA. OBJECTIVE: We aimed to classify patients with CVA using cluster analysis based on clinicophysiologic parameters and to unveil the underlying molecular pathways of these phenotypes with transcriptomic data of sputum cells. METHODS: We applied k-mean clustering to 342 newly physician-diagnosed patients with CVA from a prospective multicenter observational cohort using 10 prespecified baseline clinical and pathophysiologic variables. The clusters were compared according to clinical features, treatment response, and sputum transcriptomic data. RESULTS: Three stable CVA clusters were identified. Cluster 1 (n = 176) was characterized by female predominance, late onset, normal lung function, and a low proportion of complete resolution of cough (60.8%) after antiasthmatic treatment. Patients in cluster 2 (n = 105) presented with young, nocturnal cough, atopy, high type 2 inflammation, and a high proportion of complete resolution of cough (73.3%) with a highly upregulated coexpression gene network that related to type 2 immunity. Patients in cluster 3 (n = 61) had high body mass index, long disease duration, family history of asthma, low lung function, and low proportion of complete resolution of cough (54.1%). TH17 immunity and type 2 immunity coexpression gene networks were both upregulated in clusters 1 and 3. CONCLUSION: Three clusters of CVA were identified with different clinical, pathophysiologic, and transcriptomic features and responses to antiasthmatics treatment, which may improve our understanding of pathogenesis and help clinicians develop individualized cough treatment in asthma.

The role of HDAC2 in cigarette smoke-induced airway inflammation in a murine model of asthma and the effect of intervention with roxithromycin.

BACKGROUND: Cigarette smoke is well known to worsen asthma symptoms in asthmatic patients and to make them refractory to treatment, but the underling molecular mechanism is unclear. We hypothesized that cigarette smoke can reduce the expression of HDAC2 in asthma and the process was achieved by activating the PI3K-δ/Akt signaling pathway. We further hypothesized that roxithromycin (RXM) can alleviate the impacts by cigarette smoke. METHODS: A murine model of asthma induced by ovalbumin (OVA) and cigarette smoke has been established. The infiltration of inflammatory cells and inflammatory factors was examined in this model. Finally, we evaluated the expression of HDAC2, Akt phosphorylation levels, and the effects of RXM treatment on the model described earlier. RESULTS: Cigarette smoke exposure reduced HDAC2 protein expression by enhancing the phosphorylation of Akt in PI3K-δ/Akt signaling pathway. Furthermore, RMX reduced the airway inflammation and improved the level of expression of HDAC2 in the cigarette smoke-exposed asthma mice. CONCLUSIONS: This study provides a novel insight into the mechanism of cigarette smoke exposure in asthma and the effects of RXM treatment on this condition. These results may be helpful for treating refractory asthma and emphasizing the need for a smoke-free environment for asthmatic patients.

Two indole-2-carboxamide derivatives attenuate lipopolysaccharide-induced acute lung injury by inhibiting inflammatory response.

Acute lung injury (ALI) is the leading cause of mortality in the intensive care unit. Currently, there is no effective pharmacological treatment for ALI. In our previous study, we reported that Lg25 and Lg26, two indole-2-carboxamide derivatives, inhibited the lipopolysaccharide (LPS)-induced inflammatory cytokines in vitro and attenuated LPS-induced sepsis in vivo. In the present study, we confirmed data from previous studies that LPS significantly induced pulmonary edema and pathological changes in lung tissue, increased protein concentration and number of inflammatory cells in bronchoalveolar lavage fluids (BALF), and increased inflammatory cytokine TNF-α expression in serum and BALF, pro-inflammatory genes expression, and macrophages infiltration in lung tissue. However, pretreatment with Lg25 and Lg26 significantly attenuated the LPS-induced changes in mice. Taken together, these data indicate that the newly discovered indole-2-carboxamide derivatives could be particularly useful in the treatment of inflammatory diseases such as ALI.

Targeting myeloid differentiation protein 2 by the new chalcone L2H21 protects LPS-induced acute lung injury.

Acute inflammatory diseases are the leading causes of mortality in intensive care units. Myeloid differentiation 2 (MD-2) is required for recognizing lipopolysaccharide (LPS) by toll-like receptor 4 (TLR4), and represents an attractive therapeutic target for LPS-induced inflammatory diseases. In this study, we report a chalcone derivative, L2H21, as a new MD2 inhibitor, which could inhibit LPS-induced inflammation both in vitro and in vivo. We identify that L2H21 as a direct inhibitor of MD-2 by binding to Arg90 and Tyr102 residues in MD-2 hydrophobic pocket using a series of biochemical experiments, including surface plasmon response, molecular docking and amino acid mutation. L2H21 dose dependently inhibited LPS-induced inflammatory cytokine expression in primary macrophages. In mice with LPS intratracheal instillation, L2H21 significantly decreased LPS-induced pulmonary oedema, pathological changes in lung tissue, protein concentration increase in bronchoalveolar lavage fluid, inflammatory cells infiltration and inflammatory gene expression, accompanied with the decrease in pulmonary TLR4/MD-2 complex. Meanwhile, administration with L2H21 protects mice from LPS-induced mortality at a degree of 100%. Taken together, this study identifies a new MD2 inhibitor L2H21 as a promising candidate for the treatment of acute lung injury (ALI) and sepsis, and validates that inhibition of MD-2 is a potential therapeutic strategy for ALI.

Curcumin Analog L48H37 Prevents Lipopolysaccharide-Induced TLR4 Signaling Pathway Activation and Sepsis via Targeting MD2.

Endotoxin-induced acute inflammatory diseases such as sepsis, mediated by excessive production of various proinflammatory cytokines, remain the leading cause of mortality in critically ill patients. Lipopolysaccharide (LPS), the characteristic endotoxin found in the outer membrane of Gram-negative bacteria, can induce the innate immunity system and through the myeloid differentiation protein 2 (MD2) and Toll-like receptor 4 (TLR4) complex, increase the production of inflammatory mediators. Our previous studies have found that a curcumin analog, L48H37 [1-ethyl-3,5-bis(3,4,5-trimethoxybenzylidene)piperidin-4-one], was able to inhibit LPS-induced inflammation, particularly tumor necrosis factor α and interleukin 6 production and gene expression in mouse macrophages. In this study, a series of biochemical experiments demonstrate L48H37 specifically targets MD2 and inhibits the interaction and signaling transduction of LPS-TLR4/MD2. L48H37 binds to the hydrophobic region of MD2 pocket and forms hydrogen bond interactions with Arg(90) and Tyr(102). Subsequently, L48H37 was shown to suppress LPS-induced mitogen-activated protein kinase phosphorylation and nuclear factor κB activation in macrophages; it also dose dependently inhibits the cytokine expression in macrophages and human peripheral blood mononuclear cells stimulated by LPS. In LPS-induced septic mice, both pretreatment and treatment with L48H37 significantly improved survival and protected lung injury. Taken together, this work identified a new MD2 specific inhibitor, L48H37, as a potential candidate in the treatment of sepsis.

Anti-inflammatory effects of novel curcumin analogs in experimental acute lung injury.

BACKGROUND: Acute lung injury (ALI) and its most severe form acute respiratory distress syndrome (ARDS) have been the leading cause of morbidity and mortality in intensive care units (ICU). Currently, there is no effective pharmacological treatment for acute lung injury. Curcumin, extracted from turmeric, exhibits broad anti-inflammatory properties through down-regulating inflammatory cytokines. However, the instability of curcumin limits its clinical application. METHODS: A series of new curcumin analogs were synthesized and screened for their inhibitory effects on the production of TNF-α and IL-6 in mouse peritoneal macrophages by ELISA. The evaluation of stability and mechanism of active compounds was determined using UV-assay and Western Blot, respectively. In vivo, SD rats were pretreatment with c26 for seven days and then intratracheally injected with LPS to induce ALI. Pulmonary edema, protein concentration in BALF, injury of lung tissue, inflammatory cytokines in serum and BALF, inflammatory cell infiltration, inflammatory cytokines mRNA expression, and MAPKs phosphorylation were analyzed. We also measured the inflammatory gene expression in human pulmonary epithelial cells. RESULTS: In the study, we synthesized 30 curcumin analogs. The bioscreeening assay showed that most compounds inhibited LPS-induced production of TNF-α and IL-6. The active compounds, a17, a18, c9 and c26, exhibited their anti-inflammatory activity in a dose-dependent manner and exhibited greater stability than curcumin in vitro. Furthermore, the active compound c26 dose-dependently inhibited ERK phosphorylation. In vivo, LPS significantly increased protein concentration and number of inflammatory cells in BALF, pulmonary edema, pathological changes of lung tissue, inflammatory cytokines in serum and BALF, macrophage infiltration, inflammatory gene expression, and MAPKs phosphorylation . However, pretreatment with c26 attenuated the LPS induced increase through ERK pathway in vivo. Meanwhile, compound c26 reduced the LPS-induced inflammatory gene expression in human pulmonary epithelial cells. CONCLUSIONS: These results suggest that the novel curcumin analog c26 has remarkable protective effects on LPS-induced ALI in rat. These effects may be related to its ability to suppress production of inflammatory cytokines through ERK pathway. Compound c26, with improved chemical stability and bioactivity, may have the potential to be further developed into an anti-inflammatory candidate for the prevention and treatment of ALI.

Design, synthesis and biological evaluation of paralleled Aza resveratrol-chalcone compounds as potential anti-inflammatory agents for the treatment of acute lung injury.

Acute lung injury (ALI) is a major cause of acute respiratory failure in critically-ill patients. It has been reported that both resveratrol and chalcone derivatives could ameliorate lung injury induced by inflammation. A series of paralleled Aza resveratrol-chalcone compounds (5a-5m, 6a-6i) were designed, synthesized and screened for anti-inflammatory activity. A majority showed potent inhibition on the IL-6 and TNF-α expression-stimulated by LPS in macrophages, of which compound 6b is the most potent analog by inhibition of LPS-induced IL-6 release in a dose-dependent manner. Moreover, 6b exhibited protection against LPS-induced acute lung injury in vivo. These results offer further insight into the use of Aza resveratrol-chalcone compounds for the treatment of inflammatory diseases, and the use of compound 6b as a lead compound for the development of anti-ALI agents.

[Prognostic value of extra-vascular lung water index and pulmonary vascular permeability index in patients with ARDS].

OBJECTIVE: To investigate the correlation of the extra-vascular lung water index (EVLWI) and the pulmonary vascular permeability index (PVPI) with disease severity and their prognostic value in patients with acute respiratory distress syndrome (ARDS). METHODS: A total of 44 patients with ARDS from October 2012 to June 2014 admitted in the Second Affiliated Hospital of Wenzhou Medical University were recruited in this study. According to the severity, patients were divided into three groups (Mild group, Moderate group and Severe group); the acute physiology and chronic health evaluation system II score (APACHE II), the lung injury score (LIS), the pulse contour curve continuous cardiac output (PiCCO) and other clinical indicators were respectively monitored in the period of 24, 48, 72 hrs after admission; then the correlation of EVLWI, PVPI and oxygenation index (OI) among groups were analyzed; According to the prognosis, patients were divided into the survival group and the death group, both given the univariate and multivariate Logistic regression analysis; EVLWI, PVPI, APACHE II score, LIS and lactic acid were admitted into the receiver operating characteristic (ROC) curve analysis, and the prognosis was evaluated respectively. RESULTS: With the increase of disease severity, LIS and lactic acid gradually increased, the difference was significant among the three groups of Mild, Moderate and Severe (P<0.05). And the APACHE II score also increased gradually with the severity, but the difference was statistically significant only between the Mild group and the Severe group (P<0.01). And likewise, mild, moderate, severe ARDS patients had 1, 6, 9 cases of death, respectively. The 28-day mortality rate increased gradually after admission, with a significant difference between the Mild group and the Severe group (P<0.05). When all the 44 patients of three severities (during the 24 hrs period and during the 72 hrs period) were compared, the OI gradually decreased with the increase of severity of ARDS, while EVLWI and PVPI ascended, and differences between any two groups were statistically significant (P<0.05). In addition, there was a significant negative correlation between EVLWI and OI or between PVPI and OI (r=-0.666, -0.763, all P<0.01), and a significant positive correlation between EVLWI and PVPI, the APACHE II score or LIS (r=0.929, 0.895, 0.661, all P<0.01). Besides, OI was a predictive protection factor of ARDS, whereas lactic acid, EVLWI and PVPI were risk factors. Multivariate Logistic regression analysis showed that EVLWI and lactic acid were risk factors for ARDS death (all P<0.05). ROC curve analysis results suggested EVLWI and lactic acid were risk factors, (odd ratio (OR)> 1, and 95%CI: 1.071-5.201, 5.201-99.852, all P<0.05). CONCLUSION: EVLWI, PVPI were positively correlated with the severity of ARDS illness; EVLWI can be used as an independent risk factor for forecasting ARDS death, jointing EVLWI with PVPI could improve the accuracy of ARDS death forecasting.

[Significance of fractional exhaled nitric oxide in the diagnosis of asthma in pregnant women].

OBJECTIVE: To assess the variability of fractional exhaled nitric oxide (FeNO) and to explore the significances of FeNO in the diagnosis and treatment of asthma in pregnant women. METHODS: In a prospective study, 65 healthy pregnant women, 55 asthmatic non-pregnant women, 40 asthmatic pregnant women and 60 healthy non-pregnant women were enrolled in the study from Oct 2012 to Apr 2014.FeNO levels of the 4 groups were compared, and the variability of FeNO in different pregnancy periods of 15 healthy pregnant women were examined.We also analyzed the correlation between the level of FeNO, FEV1% pred, asthma control test (ACT) scores andblood eosinophils in patients of asthmatic pregnant women. RESULTS: The difference in the levels of FeNO between healthy pregnant women (12 ± 6) ppb and healthy non-pregnant women (14 ± 5) ppb were no significant (t = 0.508, P > 0.05) . The levels of FeNO in different pregnancy periods of healthy pregnant women were not significantly different (F = 0.656, P > 0.05). Compared with healthy pregnant women (12 ± 6) ppb, the level of FeNO was significantly higher in asthmatic pregnant patients (43 ± 21) ppb (t = 2.981, P < 0.05) , but the difference of the levels of FeNO between asthmatic non-pregnant (51 ± 32) ppb and pregnant patients (43 ± 21) ppb were no significant (t = 0.366, P > 0.05) . There was no significant correlation between FeNO level and FEV1% pred (r = 0.164, P > 0.05), nor between FeNO level and ACT scores (r = 0.272, P > 0.05) , but there was significant correlation between FeNO level and eosinophils (r = 0.723, P < 0.05). CONCLUSIONS: FeNO level is not influenced by pregnancy and different gestation periods.FeNO level in asthmatic pregnant women is increased, and FeNO is an important examination in the diagnosis of asthma during pregnancy.

Roxithromycin treatment inhibits TGF-ß1-induced activation of ERK and AKT and down-regulation of caveolin-1 in rat airway smooth muscle cells.

BACKGROUND: Roxithromycin (RXM) has been widely used in asthma treatment; however, the mechanism has not been fully understood. The aim of our study was to investigate the underlying mechanism of RXM treatment in mediating the effect of transforming growth factor (TGF)-ß1 on airway smooth muscle cells (ASMCs) proliferation and caveolinn-1 expression. METHODS: Firstly, the rat ovalbumin (OVA) model was built according to the previous papers. Rat ASMCs were prepared and cultured, and then TGF-ß1 production in ASMCs was measured by enzyme-linked immunosorbent assay (ELISA). Moreover, the proliferation of ASMCs was determined using cell counting kit (CCK-8) assay. Additionally, the expressions of caveolin-1, phosphorylated-ERK1/2 (p-ERK1/2) and phosphorylated-AKT (p-AKT) in ASMCs treated with or without PD98059 (an ERK1/2 inhibitor), wortannin (a PI3K inhibitor), ß-cyclodextrin (ß-CD) and RXM were measured by Western blot. Finally, data were evaluated using t-test or one-way ANOVA, and then a P value < 0.05 was set as a threshold. RESULTS: Compared with normal control, TGF-ß1 secretion was significantly increased in asthmatic ASMCs; meanwhile, TGF-ß1 promoted ASMCs proliferation (P < 0.05). However, ASMCs proliferation was remarkably inhibited by RXM, ß-CD, PD98059 and wortmannin (P < 0.05). Moreover, the expressions of p-ERK1/2 and p-AKT were increased and peaked at 20 min after TGF-ß1 stimulation, and then suppressed by RXM. Further, caveolin-1 level was down-regulated by TGF-ß1 and up-regulated by inhibitors and RXM. CONCLUSION: Our findings demonstrate that RXM treatment inhibits TGF-ß1-induced activation of ERK and AKT and down-regulation of caveolin-1, which may be the potential mechanism of RXM protection from chronic inflammatory diseases, including bronchial asthma.

Synthesis and evaluation of a series of novel asymmetrical curcumin analogs for the treatment of inflammation.

Curcumin has been reported to possess multiple bioactivities, such as antioxidant, anticancer, and anti-inflammatory properties, however the clinical application of curcumin has been significantly limited by its instability and poor metabolism. Modification of curcumin has led to discovery and development of lots of novel therapeutic candidates. In recent years acute and chronic inflammation has been the focus of numerous studies in various diseases. Here, we synthesized a series of asymmetrical curcumin analogs with high in vitro chemical stability, and their anti-inflammatory activity was evaluated in LPS-stimulated macrophages. According to the bio-screening results and QSAR analysis, these analogs exhibited potent activities against LPS-induced TNF-α and IL-6 release. Among the analogs of the potent anti-inflammatory activity, compounds 3b8 and 3b9 exhibited significant protection and possess enhanced anti-inflammatory activity thereby attenuated the LPS-induced septic death in mice.

[Regulatory effects of luteolin on airway inflammation in asthmatic rats].

OBJECTIVE: To explore the regulatory effects of luteolin on airway inflammation in asthmatic rats. METHODS: A total of 48 male Sprague-Dawley (SD) rats were randomly divided into 3 groups of control, asthmatic and luteolin(n = 16 each). The rat model of bronchial asthma was established in asthmatic and luteolin groups. The model was induced by intraperitoneally injecting a mixture of ovalbumin and aluminum hydroxide at Day 1 and 8. After two weeks, aomization excitation of normal saline (containing 1% ovalbumin) was induced thrice weekly. The treatment lasted 8 weeks. In control group, the mixture of ovalbumin, aluminum hydroxide and normal saline containing 1% ovalbumin was replaced by normal saline. At 30 min after aomization excitation, normal saline was given to rats in control and asthmatic groups, while 1 mg/kg luteolin was given intraperitoneally to luteolin group. The inflammatory cell number and level of interleukin-4 (IL-4) were measured in bronchoalveolar lavage fluid (BALF). The histopathological changes were observed under light microscope. The activities of peroxisome proliferator-activated receptors (PPARγ) and p38 mitogen-activated protein kinases (p38MAPK) in pulmonary tissues were detected by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The bronchial wall thickness of asthma group, along with smooth muscle thickness ((93.3 ± 7.4), (34.9 ± 2.3) µm) was more than that of control ((61.9 ± 8.2), (19.3 ± 1.5) µm) and luteolin ((76.6 ± 6.7), (25.4 ± 4.6) µm) groups (all P < 0.05). The total cell count ((5.61 ± 0.63)×10(9)/L), neutrophil count ((1.83 ± 0.09)×10(9)/L), eosinophil count ((0.59 ± 0.09)×10(9)/L) and level of IL-4 ((78.23 ± 12.73) pg/ml) in BALF of asthmatic group were markedly higher than those of control ((1.53 ± 0.31)×10(9)/L, (0.45 ± 0.21)×10(9)/L, (0.07 ± 0.03) ×10(9)/L and (21.21 ± 2.53) pg/ml) and luteolin ((3.24 ± 0.25)×10(9)/L, (1.54 ± 0.10)×10(9)/L, (0.33 ± 0.05)×10(9)/L and (43.24 ± 8.65) pg/ml) groups (all P < 0.05). The results of semi-quantitative immunohistochemical analysis showed that the p38 protein level in control group (0.143 ± 0.017) and luteolin group (0.251 ± 0.021) was significantly less than that in asthmatic group (0.362 ± 0.008) (both P < 0.01). As compared with asthmatic group, the expression of PPARγ protein markedly increased (0.247 ± 0.034) in control (0.331 ± 0.056) and luteolin (0.442 ± 0.031) groups (all P < 0.05). The level of p38 mRNA in asthmatic group (0.718 ± 0.064) was significantly higher than that of control (0.312 ± 0.052) and luteolin (0.426 ± 0.067) groups (all P < 0.01). However, the PPARγ mRNA level in asthmatic group (0.266 ± 0.036) was much less than that in control (0.573 ± 0.042) and luteolin (0.687 ± 0.054) groups (all P < 0.01). CONCLUSION: The anti-inflammatory effects of luteolin may be associated with the regulation of PPARγ expression and p38MAPK signaling pathway in asthmatic rats.

[Effects of smoking on expressions of HIF-1α and HDAC2 in asthmatic mice].

OBJECTIVE: To explore the effects of smoking on airway inflammation through the expressions of hypoxia-inducible factor-1α (HIF-1α) and histone deacetylase-2 (HDAC2) in asthmatic mice. METHODS: A total of 30 female SPF BALB/c mice were divided randomly into 3 groups of control (C), asthma (A) and smoking asthma (S). The latter two groups were sensitized and challenged with ovalbumin (OVA) for asthmatic modeling. The mice of group S were placed into a self-made fumigating box for passive smoking. While group S was sensitized and challenged with normal saline instead of OVA. The pathological changes of different groups were observed. The different cell counts of bronchoalveolar fluid (BALF) were analyzed. The expressions of HIF-1α and HDAC2 were detected by immunohistochemistry. The level of interleukin (IL)-8 in BALF was detected by enzyme-linked immunosorbent assay (ELISA). And the levels of HIF-1α and HDAC2 in lung homogenate were measured by Western blot. RESULTS: The ratios of eosinophil (EOS) to total cell numbers of BALF in groups A and S were significantly higher than that in group C ((8.90 ± 1.60)%, (7.52 ± 0.63)% vs (0.60 ± 0.10)%, both P < 0.01), while the ratio of neutrophile (NEU) in group S was higher than that in group A ((18.24 ± 5.19)% vs (8.46 ± 1.58)%, P < 0.01). Western blot showed that HIF-1α expressions in lung homogenate of groups A and S were significantly elevated than that in groups C (0.144 ± 0.008, 0.238 ± 0.015 vs 0.081 ± 0.005, both P < 0.01). While the HIF-1α level of group S was higher than that of group A (P < 0.01). And the expressions of HDAC2 in groups A and S significantly decreased than that in group C (0.287 ± 0.008, 0.164 ± 0.015 vs 0.452 ± 0.041, both P < 0.01). While the HDAC2 level of group S was lower than that of group A (P < 0.01). The BALF level of IL-8 in group S was higher than those in groups A and C ((42.07 ± 4.54) vs (21.66 ± 2.78), (14.33 ± 3.73) pg/ml, both P < 0.01). There were significantly negative correlations between the expressions of HIF-1α and HDAC2 (r = -0.950, P < 0.01) in lung as well as HDAC2 in lung and IL-8 (r = -0.855, P < 0.01) in BALF. CONCLUSION: Cigarette smoking aggravates the airway inflammation through a down-regulated expression of HDAC2 by activating HIF-1α.

Costunolide attenuates LPS-induced inflammation and lung injury through inhibiting IKK/NF-κB signaling.

Inflammation is an important pathological process of many acute and chronic diseases, such as sepsis, arthritis, and cancer. Many factors can lead to an inflammatory state of the body, among which bacterial infection plays an important role. Bacterial infection often leads to sepsis, acute lung injury (ALI), or its more serious form of acute respiratory distress syndrome, which are the main fatal diseases in intensive care units. Costunolide has been reported to possess excellent anti-inflammatory activity; however, whether it can affect inflammation induced by gram-negative bacterial is still unclear. Lipopolysaccharide (LPS) stimulated mouse peritoneal macrophages (MPMs) to release proinflammatory cytokines was used as the cell model. The mouse model of sepsis and ALI was built through injecting intravenously and intratracheally of LPS. In the present study, costunolide inhibited LPS-induced inflammatory response through IKK/NF-κB signaling pathway in macrophages. In vivo, costunolide attenuated LPS-induced septic death in mice. Meanwhile, costunolide treatment alleviated LPS-induced lung injury and inflammation via inhibiting the infiltration of inflammatory cells and the expression of inflammatory cytokines. Taken together, these results demonstrated that costunolide could attenuate gram-negative bacterial induced inflammation and diseases and might be a potential candidate for the treatment of inflammatory diseases.

Chalcone-derivative L6H21 attenuates the OVA-induced asthma by targeting MD2.

Asthma represents a significant global challenge that affects individuals across all age groups and imposes substantial social and economic burden. Due to heterogeneity of the disease, not all patients obtain benefit with current treatments. The objective of this study was to explore the impact of MD2 on the progression of asthma using L6H21, a novel MD2 inhibitor, to identify potential targets and drug candidates for asthma treatment. To establish an asthma-related murine model and evaluate the effects of L6H21, ovalbumin (OVA) was used to sensitize and challenge mice. Pathological changes were examined with various staining techniques, such as H&E staining, glycogen staining, and Masson staining. Inflammatory cell infiltration and excessive cytokine secretion were evaluated by analyzing BALF cell count, RT-PCR, and ELISA. The TLR4/MD2 complex formation, as well as the activation of the MAPK and NF-кB pathways, was examined using western blot and co-IP. Treatment with L6H21 demonstrated alleviation of increased airway resistance, lung tissue injury, inflammatory cell infiltration and excessive cytokine secretion triggered by OVA. In addition, it also ameliorated mucus production and collagen deposition. In the L6H21 treatment group, inhibition of MAPK and NF-кB activation was observed, along with the disruption of TLR4/MD2 complex formation, in contrast to the model group. Thus, L6H21 effectively reduced the formation of the MD2 and TLR4 complex induced by OVA in a dose-dependent manner. This reduction resulted in the attenuation of MAPKs/NF-κB activation, enhanced suppression of inflammatory factor secretion, reduced excessive recruitment of inflammatory cells, and ultimately mitigated airway damage. MD2 emerges as a crucial target for asthma treatment, and L6H21, as an MD2 inhibitor, shows promise as a potential drug candidate for the treatment of asthma.

Efficacy and safety of mepolizumab in a Chinese population with severe asthma: a phase III, randomised, double-blind, placebo-controlled trial.

Background: In China, the prevalence of severe asthma with eosinophilic phenotype is rising, yet treatment options are limited. Mepolizumab is the first targeted biologic therapy for eosinophilic-driven disease in China. This study (clinicaltrials.gov identifier NCT03562195) evaluated efficacy and safety of mepolizumab in Chinese patients with severe asthma. Methods: The phase III, multicentre, randomised, placebo-controlled, double-blind, parallel-group study enrolled patients aged ≥12 years with severe asthma, with two or more exacerbations in the previous year, and on inhaled corticosteroids plus at least one controller medication. Following a 1-4-week run-in, patients were randomised 1:1 to mepolizumab 100 mg or placebo subcutaneously every 4 weeks for 52 weeks. The primary end-point was annualised rate of clinically significant exacerbations (CSEs) through week 52. Secondary end-points were time to first CSE, frequency of CSEs requiring hospitalisation/emergency department visits or hospitalisation over 52 weeks, mean change in St George's Respiratory Questionnaire (SGRQ) total score and pre-bronchodilator forced expiratory volume in 1 s (FEV1) at week 52; safety was evaluated. Results: The modified intention-to-treat population included 300 patients. At week 52 with mepolizumab versus placebo, annualised rate of CSEs was 65% lower (0.45 versus 1.31 events per year; rate ratio 0.35, 95% CI 0.24-0.50; p<0.001); time to first CSE longer (hazard ratio 0.38, 95% CI 0.26-0.56; p<0.001) and number of CSEs requiring hospitalisation/emergency department visit lower (rate ratio 0.30, 95% CI 0.12-0.77; p=0.012). From baseline to week 52, SGRQ score improved (p=0.001) and pre-bronchodilator FEV1 increased (p=0.006). Incidence of adverse events was similar between treatment groups. Conclusion: Mepolizumab provided clinical benefits to patients with severe asthma in China and showed a favourable benefit-risk profile.

A Phase 1b Study of Ivonescimab, a Programmed Cell Death Protein-1 and Vascular Endothelial Growth Factor Bispecific Antibody, as First- or Second-Line Therapy for Advanced or Metastatic Immunotherapy-Naive NSCLC.

INTRODUCTION: This study (HARMONi-5) aimed to evaluate the safety and efficacy of ivonescimab (a bispecific antibody against programmed cell death protein 1 and vascular endothelial growth factor) as first- or second-line monotherapy in patients with advanced immunotherapy-naive NSCLC. METHODS: Eligible patients received intravenous ivonescimab 10 mg/kg every 3 weeks (Q3W), 20 mg/kg every 2 weeks (Q2W), 20 mg/kg Q3W, or 30 mg/kg Q3W. The primary end points were safety and objective response rate (ORR) per Response Evaluation Criteria in Solid Tumors version 1.1. RESULTS: At data cutoff (October 5, 2022), 108 patients were enrolled and received ivonescimab. Programmed death ligand-1 tumor proportion score (TPS) was greater than or equal to 1% in 74 patients (68.5%), including 35 (32.4%) with TPS greater than or equal to 50%. The median follow-up was 10.4 months (range: 8.4-10.9 mo). For all patients, ORR and disease control rate were 39.8% and 86.1%, respectively. ORR by TPS was 14.7%, 51.4%, and 57.1% in patients with TPS less than 1%, greater than or equal to 1%, and greater than or equal to 50%, respectively. In the 67 programmed death ligand-1-positive patients receiving first-line ivonescimab, the ORR was 33.3%, 52.6%, 60.0%, and 75.0% at the doses of 10 mg/kg Q3W, 20 mg/kg Q2W, 20 mg/kg Q3W, and 30 mg/kg Q3W, respectively. Grade greater than or equal to 3 treatment-related adverse events (TRAEs) were observed in 24 patients (22.2%). TRAEs leading to treatment discontinuation occurred in one patient (0.9%). TRAEs leading to death occurred in three patients (2.8%) with squamous NSCLC. The occurrence of grade greater than or equal to 3 TRAEs and grade greater than or equal to 3 bleeding events in squamous versus nonsquamous NSCLC patients was 25.5% versus 18.9% and 0.0% versus 1.9%, respectively. CONCLUSIONS: Ivonescimab monotherapy was well tolerated and found to have a promising efficacy in patients with advanced or metastatic NSCLC. CLINICALTRIALS: gov identifier: NCT04900363.

[Functional role of caveolin-1 in airway smooth muscle cells proliferation and regulatory effect of roxithromycin].

OBJECTIVE: To explore the functional role of caveolin-1 in airway smooth muscle cells (ASMCs) proliferation and examine the regulatory effect of roxithromycin. METHODS: The rat model of bronchial asthma was established. Electron microscope was employed to observe the status of caveolae and light microscope for the histological changes in pulmonary tissues. The primarily cultured ASMCs were divided into 5 groups: control (group A), asthmatic ASMCs (group B), PD98059 (group C), roxithromycin (group D) and methyl-ß-cyclodextrin (group E). Cell proliferation was detected by Cell Counting Kit-8 (CCK-8). And the expressions of caveolin-1, extracellular regulated protein kinases (ERK) and monocyte chemotactic protein (MCP)-1 were detected by Western blot and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The cell proliferation of asthmatic ASMCs (0.68 ± 0.15, 0.63 ± 0.13) in groups C and D were significantly less than those in group B (0.96 ± 0.14) (both P < 0.05) while group E was more than group B (1.26 ± 0.11 vs 0.96 ± 0.14, P < 0.05). The content of caveolin-1 (0.392 ± 0.064, 0.332 ± 0.057) in groups C and D were higher than those in group B (0.237 ± 0.032) (both P < 0.05) while ERK1/2 protein level in groups C and D (0.241 ± 0.017, 0.268 ± 0.007) were less than those in group B (0.346 ± 0.009) (both P < 0.01). And MCP-1 protein level in groups C and D (0.198 ± 0.015, 0.286 ± 0.019) were less than those in group B (0.482 ± 0.026) (both P < 0.01). The ERK mRNA level in groups C and D (0.277 ± 0.043, 0.338 ± 0.026) were less than those in group B (0.591 ± 0.022) (both P < 0.01). And also MCP-1 mRNA in groups C and D (0.212 ± 0.042, 0.249 ± 0.032) were less than those in group B (0.676 ± 0.053) (all P < 0.01) CONCLUSIONS: Caveolin-1 preventing the proliferation of asthmatic ASMCs is most likely mediated by ERK1/2 signal pathway and a down-regulation of MCP-1 expression. And roxithromycin reduces the proliferation of asthmatic ASMCs through up-regulating the expression of caveolin-1 and inhibiting the expression of MCP-1.

Corrigendum to "Clinical Characteristics of Asymptomatic Patients with SARS-CoV-2 in Zhejiang: An Imperceptible Source of Infection".

[This corrects the article DOI: 10.1155/2020/2045341.].