[Inhibition of M2 macrophage polarization and reduction of airway inflammation in asthmatic mice with lncRNA MRAK088388 knockout].

Objective To investigate the long non-coding RNA(lncRNA) MRAK08838 regulates macrophage function to influence the development of asthmatic airway inflammation. Methods MRAK088388 gene knockout (MRAK088388-/-) mouse model was prepared and allergic asthma was induced by dust mite protein Dermatophagoides farinae 1 (Der f1). The mice were sacrificed after 28 days of modeling, and serum was collected to measure IgE and IgG. The FinePointe RC system was used to measure airway hyperresponsiveness and evaluate lung function in mice. Lung tissue was taken for HE staining, and periodic acid-Schiff (PAS) staining was used to evaluate inflammatory infiltration and mucus secretion in mouse lungs. Fluorescence quantitative PCR was used to detect the expression level of lncRNA MRAK08838 in bronchoalveolar lavage fluid (BALF) cells and lung tissue of asthmatic mice. ELISA was used to detect the levels of inflammatory cytokines IFN-γ, IL-4, IL-5, IL-13, IL-10 and IL-17A. Flow cytometry was used to evaluate the phenotype of macrophages in BALF and lung tissue, as well as the proportion of neutrophils, eosinophils, and alveolar macrophages. The changes of the above indicators were detected in mice by adoptive transfer of bone marrow-derived macrophages (BMDM). Results Under the challengle of Der f1, MRAK088388-/- mice showed reduced allergic airway inflammation, including reduced eosinophils in BALF and reduced production of IgE and IgG1. In addition, Der f1-treated MRAK088388-/- mice had fewer M2 macrophages than wild-type asthmatic mice. Wild-type mouse BMDM (M0) and Der f1-treated MRAK088388-/- mice also showed mild inflammatory response. Conclusion Knockout of MRAK088388 alleviates airway inflammation in asthmatic mice by inhibiting M2 polarization of airway macrophages.

Linc00511 Knockdown Inhibited TGF-ß1-Induced Epithelial-Mesenchymal Transition of Bronchial Epithelial Cells by Targeting miR-16-5p/Smad3.

BACKGROUND: Airway remodeling in patients with asthma was correlated with induced epithelial-mesenchymal transition (EMT) of bronchial epithelial cells. OBJECTIVE: This study examined the mechanism of Linc00511 on induced EMT of bronchial epithelial cells after transforming growth factor-ß1 (TGF-ß1) induction. METHODS: The human bronchial epithelial cell 16HBE was treated with 10 ng/mL TGF-ß1 for 12 h, 24 h, or 48 h to induce EMT. Cell proliferation and migration rate were detected using CCK8 and wound healing assays, respectively. The expression of key markers of EMT (E-cadherin, N-cadherin, Small mothers against decapentaplegic family member 3 [Smad3], and slug) was tested by Western blot. RESULTS: We found that Linc00511 was time dependently increased in TGF-ß-treated 16HBE cells. Silencing Linc00511 reduced 16HBE cell proliferation, migration, and EMT progress. In addition, the dual-luciferase reporter assay showed Linc00511 was a molecular sponge for miR-16-5p. MiR-16-5p decreased the expression of Smad3 by targeting its 3'-untranslated region (3'UTR). After TGF-ß1 exposure, miR-16-5p silencing counteracted the decreases of 16HBE cell proliferation, migration, and EMT induced by Linc00511 knockdown. And Smad3 overexpression also reversed the inhibitory effect of Linc00511 knockdown on proliferation, migration, and EMT progression in TGF-ß1-induced human bronchial epithelial cells. CONCLUSION: Linc00511 may be a valuable biomarker for asthma therapy.

Interspecific interactions alter plant functional strategies in a revegetated shrub-dominated community in the Mu Us Desert, China.

BACKGROUND AND AIMS: Previous studies investigating plant-plant interactions have focused on plant growth, context dependence and shifts in interactive outcomes. However, changes in functional traits in the context of interactions have been inadequately explored; few studies have focused on the effects of interactions on the plasticity of functional strategies. METHODS: We conducted a 4-year removal experiment for the xeric shrub Artemisia ordosica and perennial graminoids (PGs) in the Mu Us Desert, northern China. Soil nutrient content, biomass and 12 functional traits related to plant morphology and nutrient status were measured for the shrub species and a dominant PG species (i.e. Leymus secalinus) in the presence and absence of shrubs and PGs. KEY RESULTS: Shrubs affected the functional traits of L. secalinus, reducing leaf dry matter content and increasing plant height, which probably promoted the functional strategy of L. secalinus towards a more resource-acquisitive and competitive strategy. In contrast, when the shrubs were affected by PGs, they shifted towards a resource-conservative and stress-tolerative strategy, by increasing leaf dry matter content and decreasing specific leaf area. Moreover, the shrub species relied more on internal nutrient recycling (higher nitrogen resorption efficiency) rather than on external nitrogen uptake under nitrogen competition; instead, L. secalinus tended to exhibit higher external nitrogen uptake from soil during nitrogen shortages. CONCLUSIONS: This study indicated that the functional strategies and nutrient cycling of the shrub species and the dominant PG were altered by each other. The shifts in functional traits may help plants to coexist in the community for a relatively long time. Our findings highlighted that interspecific interactions alter plant functional strategies and provided new insights into community assembly and succession mechanisms in a revegetated shrubland for ecological restoration of drylands.

N enrichment, increased precipitation, and the effect of shrubs collectively shape the plant community in a desert ecosystem in northern China.

Understanding the responses of biological communities to global climate change is pivotal to accurately forecasting future dynamics and developing effective strategies for the adaptive ecological management of desert ecosystems. Although direct demographic responses of plant species to climatic factors have been widely acknowledged, they are also regulated by interspecific interactions (i.e., the effects of shrubs on herbaceous plants). The magnitude and direction of regulation of such interspecific interactions remain unclear. To address this knowledge gap, a full factorial field experiment simulating three levels of N enrichment (ambient, 10 kg N ha-1 yr-1, and 60 kg N ha-1 yr-1) and three levels of precipitation (ambient, 20% increase, and 40% increase) were conducted in the Mu Us Desert, northern China. N enrichment and increased precipitation significantly increased herbaceous productivity by improving the soil water content and nutrient availability (e.g., soil DIN:SAP) when shrubs were not present. Taller species responded to N enrichment, whereas those with a greater specific leaf area responded to increased precipitation. When shrubs were present, they acted as a 'buffer islands' that moderated the responses of herbaceous species to N enrichment and increased precipitation by weakening the effect of the improved soil water status. The magnitude of the effect of shrubs on herbaceous biomass and richness was comparable to that of N enrichment and increased precipitation. Our results highlight the importance and complexity of both large-scale environmental changes and small-scale interspecific interactions in structuring plant communities in desert ecosystems. Moreover, abiotic environmental factors and biotic interactions should be integrated in efforts to predict the responses of plant communities to future global change in desert ecosystems.

Fixed Ratio Versus Lower Limit of Normal: Health Status and Risk Factors for COPD Overdiagnosis.

BACKGROUND: The threshold of the lower limit of the normal range of lung function has been suggested to be more accurate than the 0.7 fixed ratio (FEV1/FVC < 0.7) for a diagnosis of COPD. We aimed to explore the health status and risk factors of patients overdiagnosed with COPD when using the lower limit of the normal range as a diagnostic reference. METHODS: Subjects with COPD diagnosed by a pulmonologist according to guidelines of the Global Initiative for Chronic Obstructive Lung Disease were recruited from October 2016 to April 2018. Overdiagnosed COPD was defined as FEV1/FVC that meets the criterion of the 0.7 fixed ratio but not the the lower limit of the normal range criterion. Spirometry and questionnaires were performed by eligible subjects. RESULTS: Of the 513 subjects included in the final analysis, 20 (3.9%) were overdiagnosed when using the lower limit of the normal range as the diagnostic reference. The subjects who were overdiagnosed were older, weighed more, had better lung function, lower modified Medical British Research Council scores, and higher St. George's Respiratory Questionnaire and 36-item Short Form Survey scores than the subjects who were correctly diagnosed. Older age, heavier weight, exposure to cooking oil fumes, or a new-built or newly renovated home were associated with an increased risk of overdiagnosis of COPD (age adjusted odds ratio (OR) 1.17, 95% CI 1.09-1.26; weight adjusted OR 1.08, 95% CI 1.03-1.13; exposure to cooking oil fumes adjusted OR 3.00, 95% CI, 1.04-8.68; exposure to new-built or newly renovated home adjusted OR 10.88, 95% CI 1.46-80.87. CONCLUSIONS: The subjects with overdiagnosed COPD had a better health status and lung function than the subjects who were correctly diagnosed. Older age, heavier weight, and exposure to cooking oil fumes or a new-built or newly renovated home were factors associated with the overdiagnosis of COPD. These findings may help reduce overdiagnosis of COPD.

Nebulized Inhalation of Anti-Nerve Growth Factor Microspheres Inhibits Airway Remodeling in an Ovalbumin-Induced Rat Asthma Model.

BACKGROUND: Airway remodeling is considered an important factor in refractory and uncontrollable asthma. Previous studies have confirmed that anti-nerve growth factor (NGF) antibody can ameliorate airway remodeling. However, whether nebulized inhalation of anti-NGF microspheres (NANM) can inhibit airway remodeling is not clear. The purpose of this study was to investigate the effects of NANM on ovalbumin (OVA)-induced airway remodeling, and the mechanisms involved. METHODS: Anti-NGF microspheres were produced using a polymer alloy method. OVA was used to establish a rat model of asthma airway remodeling. Rats were treated with inhalation atomized anti-NGF antibody or NANM. Airway inflammation, airway reactivity, and airway remodeling were measured. Lung tissue P-Smad3 and tumor growth factor (TGF)-ß1 mRNA and protein expression were also measured. RESULTS: The anti-NGF antibody microsphere encapsulation rate was high, and the release time was long. NANM markedly attenuated OVA-induced airway remodeling, such as collagen deposition, average pulmonary resistance, the WAm/Pbm, WAt/Pbm, and Wcol/Pbm ratios (WAt, bronchial wall area; Pbm, perimeter of basement membrane; WAm, smooth muscle wall area; Wcol, airway collagen fiber area). Compared with the anti-NGF antibody group and the OVA group, the expression of TGF-ß1 mRNA, TGF-ß1 protein, and P-Smad3 in the NANM group were markedly decreased. CONCLUSIONS: NANM ameliorated OVA-induced airway remodeling, partly through regulation of the TGF-ß1/Smad3 pathway.

Nitrogen enrichment alters nutrient resorption and exacerbates phosphorus limitation in the desert shrub Artemisia ordosica.

Increasing nitrogen (N) deposition and precipitation are major drivers of global changes that are expected to influence plant nutrient resorption in desert ecosystems, where plant growth is often nutrient and water limited. However, knowledge on the effects of increased N and precipitation on them remain poorly understood. This study determined the effects of increased N (ambient, 60 kg N ha-1 year-1) and water supply (ambient, +20%, +40%), and their combination on the leaf nutrient resorption of Artemisia ordosica, a dominant shrub in the Mu Us Desert of northern China. After 2 years of treatments, only N addition significantly decreased the N resorption efficiency of A. ordosica. Both N and water addition had no effect on the phosphorus (P) resorption efficiency of this shrub, and there were no interactive effects of N and water availability on shrub nutrient resorption. The responses of shrub leaf N:P ratio tended to saturate as soil available N:P increased. The aboveground net primary productivity of A. ordosica was positively correlated with leaf P resorption efficiency, rather than N resorption efficiency. Our results suggest that N addition exacerbated the P limitation of the shrub growth and played a more fundamental role than water addition in controlling the nutrient resorption process of the desert shrub A. ordosica. This information contributes to understand the relationship between nutrient conservation strategy and plant growth of desert shrub species under global environmental changes.

Resource Availability Drives Responses of Soil Microbial Communities to Short-term Precipitation and Nitrogen Addition in a Desert Shrubland.

Desert microbes are expected to be substantially sensitive to global environmental changes, such as precipitation changes and elevated nitrogen deposition. However, the effects of precipitation changes and nitrogen enrichment on their diversity and community composition remain poorly understood. We conducted a field experiment over 2 years with multi-level precipitation and nitrogen addition in a desert shrubland of northern China, to examine the responses of soil bacteria and fungi in terms of diversity and community composition and to explore the roles of plant and soil factors in structuring microbial communities. Water addition significantly increased soil bacterial diversity and altered the community composition by increasing the relative abundances of stress-tolerant (dormant) taxa (e.g., Acidobacteria and Planctomycetes); however, nitrogen addition had no substantial effects. Increased precipitation and nitrogen did not impact soil fungal diversity, but significantly shifted the fungal community composition. Specifically, water addition reduced the relative abundances of drought-tolerant taxa (e.g., the orders Pezizales, Verrucariales, and Agaricales), whereas nitrogen enrichment decreased those of oligotrophic taxa (e.g., the orders Agaricales and Sordariales). Shifts in microbial community composition under water and nitrogen addition occurred primarily through changing resource availability rather than plant community. Our results suggest that water and nitrogen addition affected desert microbes in different ways, with watering shifting stress-tolerant traits and fertilization altering copiotrophic/oligotrophic traits of the microbial communities. These findings highlight the importance of resource availability in driving the desert microbial responses to short-term environmental changes.

Airway reversibility in asthma and phenotypes of Th2-biomarkers, lung function and disease control.

BACKGROUND: High bronchodilator reversibility in adult asthma is associated with distinct clinical characteristics. In this study, we aim to make a comparison with T-helper 2 (Th2)-related biomarkers, lung function and asthma control between asthmatic patients with high airway reversibility (HR) and low airway reversibility (LR). METHODS: Patients with asthma diagnosed by pulmonologist according to Global Initiative for Asthma guidelines were recruited from the outpatient department of our hospital from August 2014 to July 2017. Patients were divided into HR and LR subgroups based on their response to bronchodilators of lung function (HR = Δforced expiratory volume in one second (FEV1) postbronchodilator ≥ 20%). Blood eosinophil count and serum IgE level, which are biomarkers of T-helper (Th)-2 phenotypes, were detected for patients. Asthma Control Test (ACT) was used to assess asthma control after the first-month initial treatment. RESULTS: A total of 265 patients with asthma were followed 1 month after initial treatment. HR group shows a higher level of Th2-high biomarkers (blood eosinophil count (10^9/L): 0.49 ± 0.28 vs 0.36 ± 0.19, P < 0.01; IgE (ng/ml): 1306 ± 842 vs 413 ± 261, P < 0.01), lower baseline lung function (FEV1%pred: 51.91 ± 19.34% vs 60.42 ± 19.22%, P < 0.01; forced expiratory flow (FEF)25-75: 0.76 ± 0.37 vs 1.00 ± 0.67, P < 0.01; FEF25-75%pred: 21.15 ± 10.09% vs 29.06 ± 16.50%, P < 0.01), and better asthma control (ACT score: 22 ± 4 vs 20 ± 4, P = 0.01) than LR group. HR was associated with a decreased risk of uncontrolled asthma after the first-month initial treatment (adjusted OR: 0.12 [95% confidence intervals: 0.03-0.50]). CONCLUSIONS: HR is a physiologic indicator of lower lung function and severer small airway obstruction, and is more related with an increased level of Th2-biomarkers than LR. Moreover, HR may indicate controlled asthma after the first-month initial treatment. This finding may contribute to identification of asthma endotype.

Plasticity in Meristem Allocation as an Adaptive Strategy of a Desert Shrub under Contrasting Environments.

The pattern of resource allocation to reproduction vs. vegetative growth is a core component of a plant's life-history strategy. Plants can modify their biomass allocation patterns to adapt to contrasting environments. Meristems can have alternative fates to commit to vegetative growth, reproduction, or remaining inactive (dormant or senescent/dead). However, knowledge about whether meristem fates can interpret adaptive changes in biomass allocation remains largely unknown. We measured aboveground plant biomass (a proxy of plant size) and meristem number of a dominant shrub Artemisia ordosica in three populations occupying different habitats in the Mu Us Desert of northern China. Size-dependent biomass allocation and meristem allocation among habitats were compared. The size-dependent biomass allocation and meristem allocation of A. ordosica strongly varied across habitats. There were significant positive linear relationships between meristem allocation and biomass allocation in all habitats, indicating that meristem allocation is an indicator of the estimated resource allocation to reproductive and vegetative organs in this species. Plasticity in meristem allocation was more likely caused by larger individuals having less active meristems due to environmental stress. Vegetative meristems (VM) were likely more vulnerable to environmental limitation than reproductive ones, resulting in the ratio of resource investment between vegetative and reproductive functions exhibiting plasticity in different habitats. A. ordosica invested a higher fraction of its resource to reproduction in the adverse habitat, while more resource to vegetative growth in the favorable habitat. A. ordosica adopts different resource allocation patterns to adapt to contrasting habitat conditions through altering its meristem fates. Our results suggest that the arid-adapted shrub A. ordosica deactivates more VM than reproductive ones to hedge against environmental stress, representing an important adaptive strategy. This information contributes to understand the life-history strategies of long-lived plants under stressful environments.

Increased Precipitation and Nitrogen Alter Shrub Architecture in a Desert Shrubland: Implications for Primary Production.

Shrublands are one of the major types of ecosystems in the desert regions of northern China, which is expected to be substantially more sensitive to global environmental changes, such as widespread nitrogen enrichment and precipitation changes, than other ecosystem types. However, the interactive effects of nitrogen and precipitation on them remain poorly understood. We conducted a fully factorial field experiment simulating three levels of precipitation (ambient, +20%, +40%) and with two levels of nitrogen deposition (ambient, 60 kg N ha-1 yr-1) in a desert shrubland in the Mu Us Desert of northern China. We used plant architectural traits (plant cover, volume, twig size and number) as proxies to predict aboveground net primary productivity (ANPP) of the dominant shrub (Artemisia ordosica Krasch), and assessed the responses of plant productivity and architectural traits to water and nitrogen addition. We found significant differences in twig size and number of A. ordosica under water and nitrogen treatments but not in shrub cover/volume, which suggest that twig size and number of the shrub species were more sensitive to environmental changes. The productivity of the overall community was sensitive to increased precipitation and nitrogen, and shrubs played a more important role than herbaceous plants in driving productivity in this ecosystem. Precipitation- and nitrogen-induced increases in vegetation production were positively associated with increases in twig size and number of the dominant shrub. Water addition enhanced the twig length of A. ordosica, while nitrogen addition resulted in increased twig density (the number of twigs per square meter). Water and nitrogen interacted to affect twig length, but not twig number and shrub ANPP. The trade-off, defined as negative covariance between twig size and number, was likely the mechanism underlying the responses of twig length and shrub ANPP to water and nitrogen interactions. Our results highlight the sensitivity of twig size and number as indicators to estimate shrub production and the mechanism underpinning desert shrub ANPP response to global environmental changes.

Patterns and possible mechanisms of soil CO2 uptake in sandy soil.

It has been reported that soils in drylands can absorb CO2, although the patterns and mechanisms of such a process remain under debate. To address this, we investigated the relationships between soil CO2 flux and meteorological factors and soil properties in Northwest China to reveal the reasons for "anomalous" soil CO2 flux in a desert ecosystem. Soil CO2 flux increased significantly and exponentially with surficial turbulence at the diel scale under dry conditions (P<0.05), whereas the relationship under wet conditions was insignificant. Furthermore, soil CO2 flux demonstrated remarkable negative correlation with soil air pressure (P<0.05) in both dry and wet conditions. Analysis considering Henry's Law indicated that soil water content was insufficient to dissolve the absorbed CO2 in dry conditions, but was sufficient in wet conditions. The concentration of soil HCO3(-) in the morning was higher than in the evening in dry conditions, but this pattern was reversed in wet conditions. These results imply that CO2 outgassing induced by turbulence, expansion of soil air, CO2 effusion from soil water, and carbonate precipitation during daytime can explain the abiotic diurnal CO2 release. Moreover, CO2 pumping from the atmosphere into the soil, caused mainly by carbonate dissolution, can account for nocturnal CO2 absorption in dry conditions. The abiotic soil CO2 flux pattern (CO2 absorption throughout the diel cycle) in wet conditions can be attributed to downward mass flow of soil CO2 and intensified soil air shrinkage, CO2 dissolving in soil water, and carbonate dissolution. These results provide a basis for determining the location of abiotic fixed carbon within soils in desert ecosystems.

Effects of N-linked glycosylation of the fusion protein on replication of human metapneumovirus in vitro and in mouse lungs.

The fusion (F) protein is an important membrane glycoprotein necessary for cellular entry and replication of human metapneumovirus (hMPV). Selective prevention of N-linked glycosylation may compromise the catalytic and fusion functions of the F protein. By using site-directed mutagenesis and reverse genetics, recombinant mutant viruses lacking one or two N-linked glycosylation sites in the F protein were constructed. M1, which lacked glycosylation at position 57 of the F protein, had slightly compromised replication, whereas M2 and M4, which lacked glycosylation at position(s) 172 or 57 and 172, respectively, showed profound impairment of replication when compared with wild-type (WT) NL/1/00-GFP virus in both Vero E6 cells and mouse lungs. M2 was less fit than WT virus in vitro and in immunocompromised mouse lungs. The F proteins of WT and mutant viruses were similarly expressed on the infected cell membrane, while the activated fusion protein subunits, F1 of M2 and M4, were produced in lower quantities compared with those of WT and M1 virus. The mutated viruses lacking N-linked glycosylation at position 353, either individually or together with other sites, could not be recovered. Thus, N-linked glycosylation may be involved in the catalysis of the fusion protein from F0 to F1 and F2, which is critical for fusion function. Strategies targeting N-linked glycosylation may be helpful for developing attenuated live vaccines or antiviral drugs for hMPV.

[Effects of vitamin C on A549 cell proliferation, apoptosis and expressions of Caspase, Survivin].

BACKGROUND AND OBJECTIVE: It was proven that Vitamin C could inhibit the growth of many types of tumors as an antioxidant. The aim of this study is to explore role of Vitamin C in proliferation and apoptosis of lung carcinoma cell line A549 and the underlying mechanism. METHODS: A549 cells were cultured in vitro and incubated with Vitamin C. The cell viability was measured by growth curve and clonogentic assay. Flow cytometry was used to analyze cell cycle and detect apoptosis. The levels of expression of Caspase-3 mRNA and Survivin mRNA were detected by RT-PCR. RESULTS: Vitamin C of 400 microg/mL, 4 mg/mL significantly inhibited the growth of A549 cell lines (P = 0.024, P = 0.015, respectively). Flow cytometry showed that the cells major stagnation stayed in the G0/G1 and S phase and the apoptotic rate increased with time prolonged. Vitamin C signifiantly up-regulated the expression of Caspase-3 mRNA, but had no effect on Survivin mRNA. CONCLUSION: Vitamin C can inhibit the proliferation of A549, block A549 cells in G0/G1 and S phase, and induce apoptosis of A549 cells. Apotosis occurred by up-regulated the expression of Caspase-3.