Extracellular Adenosine Diphosphate Stimulates CXCL10-Mediated Mast Cell Infiltration Through P2Y1 Receptor to Aggravate Airway Inflammation in Asthmatic Mice.

Asthma is an inflammatory disease associated with variable airflow obstruction and airway inflammation. This study aimed to explore the role and mechanism of extracellular adenosine diphosphate (ADP) in the occurrence of airway inflammation in asthma. The expression of ADP in broncho-alveolar lavage fluid (BALF) of asthmatic patients was determined by enzyme linked immunosorbent assay (ELISA) and the expression of P2Y1 receptor in lung tissues was determined by reverse transcription-quantitative polymerase chain reaction. Asthmatic mouse model was induced using ovalbumin and the mice were treated with ADP to assess its effects on the airway inflammation and infiltration of mast cells (MCs). Additionally, alveolar epithelial cells were stimulated with ADP, and the levels of interleukin-13 (IL-13) and C-X-C motif chemokine ligand 10 (CXCL10) were measured by ELISA. We finally analyzed involvement of NF-κB signaling pathway in the release of CXCL10 in ADP-stimulated alveolar epithelial cells. The extracellular ADP was enriched in BALF of asthmatic patients, and P2Y1 receptor is highly expressed in lung tissues of asthmatic patients. In the OVA-induced asthma model, extracellular ADP aggravated airway inflammation and induced MC infiltration. Furthermore, ADP stimulated alveolar epithelial cells to secrete chemokine CXCL10 by activating P2Y1 receptor, whereby promoting asthma airway inflammation. Additionally, ADP activated the NF-κB signaling pathway to promote CXCL10 release. As a "danger signal" extracellular ADP could trigger and maintain airway inflammation in asthma by activating P2Y1 receptor. This study highlights the extracellular ADP as a promising anti-inflammatory target for the treatment of asthma.

Breaking the Summer Dormancy of Pinellia ternata by Introducing a Heat Tolerance Receptor-Like Kinase ERECTA Gene.

Pinellia ternata is a perennial traditional Chinese medicinal plant that undergoes different phenological patterns of dormancy depending on where it is growing. Plants grown in central and southern China typically display two growth cycles every year before and after hot summer days, exhibiting a summer dormancy. However, germplasms from these areas do not go into a dormancy phase in northern China where the summer monthly average temperatures range from 29-31°C. The northern China herbal growers prefer plant stocks from central China due to their longer growing quality and better tuber harvests. Here, we introduced a heat responsive receptor-like kinase ERECTA (ER) gene into P. ternata to explore changes in the growth cycle which were aimed at disrupting the summer dormancy. The 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene was also co-transformed with ER to improve the commercial trait. For the thermo-tolerance evaluation, all plants were treated with high temperatures (35°C/40°C) in a growth chamber or grown in natural field temperature in an isolated field before measurement of different agricultural, biochemical and physiological traits. The transgenics showed significantly (P < 0.05) higher heat tolerance, maintaining healthy vegetative growth unlike the empty vector (EV) harboring controls that became chlorotic and necrotic. Better performance in some of the monitored physiological traits was evident for overexpression lines exposed to the heat stress. In open isolated field trials, the transgenic genotypes did not show a summer dormancy but had a survival rate of 84-95%. The tuber biomasses were also significantly (P < 0.05) higher for the transgenic lines as compared to the EV controls, except for line ER118. Metabolites analysis indicated that the HMGR overexpressing lines (HMGR orHMGR + ER) exhibited significantly higher amounts of bioactive compounds including aromadendrene-4, 10-diol and 4, 8, 13-cyclotetradecatriene-1, 3-diol, 1, 5, 9-trimethyl-12-(1-methylethyl). Our findings show that the summer dormancy of P. ternata which is a naturally evolved trait, can be removed by a single heat responsive gene. The study contributes to generating heat tolerant new Pinellia varieties with enhanced commercially valuable chemicals.

Circular RNA circABCC4 regulates lung adenocarcinoma progression via miR-3186-3p/TNRC6B axis.

Lung adenocarcinoma (LUAD), a general kind of bronchogenic malignancy globally, is depicted as one of the most critical factors affecting human health severely. Featured with loop structure, circular RNA (circRNA) has been described as an essential regulator of multiple human malignancies. Nevertheless, knowledge concerning the regulatory function of circRNA in LUAD progression remains limited. Identified as a novel circRNA, circABCC4 has not been studied in LUAD as yet. This is the first time to probe into the underlying role of circABCC4 in LUAD. In this study, a notably elevated expression of circABCC4 was found in LUAD tissues and cells. Besides, circABCC4 is verified to be characterized with a circular structure in LUAD. Functional assays elucidated that knockdown of circABCC4 significantly impaired LUAD cell proliferation, migration as well as accelerated cell apoptosis. Molecular mechanism experiments later revealed that circABCC4 could bind with miR-3186-3p and miR-3186-3p was a tumor suppressor in LUAD. Moreover, TNRC6B was validated to combine with miR-3186-3p, and its expression was respectively negatively and positively regulated by miR-3186-3p and circABCC4 in LUAD. Final rescue experiments further delineated that TNRC6B upregulation partially restored circABCC4 downregulation-mediated effect on LUAD progression. In sum, circABCC4 regulates LUAD progression via miR-3186-3p/TNRC6B axis.

LncRNA BLACAT1 is involved in chemoresistance of non­small cell lung cancer cells by regulating autophagy.

The aim of the present study was to determine the effect of the long non­coding RNA (lncRNA) bladder cancer­associated transcript 1 (BLACAT1) in chemoresistance of non­small cell lung cancer (NSCLC) cells. Expression of lncRNA BLACAT1, microRNA (miR)­17, autophagy­related protein 7 (ATG7), multidrug­resistance protein 1 (MRP1), and the autophagy­associated proteins light chain 3 (LC3)­II/LC3­I and Beclin 1 were detected using the reverse transcription­quantitative polymerase chain reaction and western blot analysis. Cell viability was determined using an MTT assay. The interaction between BLACAT1 and miR­17 was determined using RNA immunoprecipitation and RNA pull­down assays. A cisplatin (DDP)­resistant NSCLC cell A549/DDP xenograft model in nude mice was established to investigate the effect of BLACAT1 on the chemoresistance of NSCLC cells. Compared with in DDP­sensitive NSCLC cells, expression of BLACAT1, ATG7, MRP1, LC3­II/LC3­I and Beclin 1 was significantly upregulated in DDP­resistant NSCLC cells, whereas miR­17 was downregulated in DDP­resistant NSCLC cells. Short interfering RNA against BLACAT1 decreased the viability of DDP­resistant NSCLC cells. In addition, BLACAT1 interacted with miR­17, and negatively regulated miR­17. BLACAT1 promoted ATG7 expression through miR­17, and facilitated autophagy and promoted chemoresistance of NSCLC cells through miR­17/ATG7. Finally, in vivo experiments indicated that inhibition of BLACAT1 ameliorated the chemoresistance of NSCLC. BLACAT1 was upregulated in DDP­resistant NSCLC cells, and promoted autophagy and chemoresistance of NSCLC cells through the miR­17/ATG7 signaling pathway.

Improved ganoderic acids production in Ganoderma lucidum by wood decaying components.

Ganoderma lucidum is a legendary Traditional Chinese Medicine (TCM) over a few thousands of years and one kind of its major active components are Ganoderic acids (GAs). GAs are largely produced in the mushroom primordium and fruiting body but much less in mycelium stage. However, little is known on the underlying regulatory mechanism. As a saprophytic fungus, G. lucidum solely obtains nutrients by wood decaying. Wood in general contains sophisticated chemical components with diverse structural units. To explore a strategy that extensively leads to GAs induction in the submerged liquid fermentation, all chemical components that might be possibly from the wood decaying were tested individually as GAs inducers. It was found that GAs production increased 85.96% by 1.5% microcrystalline cellulose (MCC) and 63.90% by 0.5% D-galactose. The transcription level of a few rate-limiting or chemically diverting enzymes responsible for GAs biosynthesis was greatly induced by MCC and D-galactose. The concentration and time-course titration study indicated that these two chemicals might not be utilized as carbon sources but they played a comprehensive role in the secondary metabolites synthesis. Our data indicated that MCC and D-galactose might be further industrialized for higher GAs production in G. lucidum in submerged fermentation.

Erythromycin enhances the anti-inflammatory activity of budesonide in COPD rat model.

Glucocorticoids (GCs) have been widely applied to treat patients with chronic obstructive pulmonary disease (COPD). But the effect of GCs was not ideal. This study was to observe whether erythromycin could enhance the anti-inflammatory activity of budesonide in COPD model rats and to explore the mechanism involved. In this study, male Sprague-Dawley rats were divided into five groups: healthy control group (H group), COPD model group (C group), erythromycin group (E group), budesonide group (B group) and erythromycin + budesonide group (E+B group). The rats in groups of C, E, B and E+B were developed into COPD models. Different groups were given different drug interventions. The levels of 8-iso-PGF2α, IL-8, and TNF-α in BALF and serum were measured with ELISA. The protein expression levels of HDAC2, PI3K, and p-AKT in lung tissue were measured with Western-blot and immunohistochemistry. The levels of 8-iso-PGF2α, IL-8, and TNF-α in BALF and serum were lower in E+B group than those in B group and C group (all P<0.001).The protein expression level of HDAC2 was higher and PI3K and p-AKT were lower in E+B group than those in B group and C group (all P<0.001). Moreover, the expression levels of HDAC2 were negatively correlated with the levels of 8-iso-PGF2α, IL-8 and TNF-α both in serum and BALF and the expression levels of PI3K and p-AKT among the five groups, with all P<0.001. We conclude that erythromycin can enhance the anti-inflammatory activity of budesonide in COPD model rats, possibly through inhibiting the PI3K/AKT pathway and enhancing the activity of HDAC2.

MiR-449c targets c-Myc and inhibits NSCLC cell progression.

MicroRNAs (miRNA) play an important role in tumorigenesis, proliferation, and differentiation. Altered miRNA expression in cancer indicates that miRNAs can function as tumor suppressors or oncogenes. MiR-449c downregulation in non-small cell lung cancer (NSCLC) compared with normal lung tissues was investigated in this study. NSCLC cell proliferation and invasion assays indicate that transfection of miR-449c expression plasmid inhibits the proliferation and invasion ability of NCI-H23 and NCI-H838 cells. In addition, miR-449c overexpression could suppress tumor growth in vivo. Morever, c-Myc was identified as a direct target gene of miR-449c. These findings clearly suggest that miR-449c downregulation and c-Myc amplification may be involved in the development of NSCLC.