Scorpio and centipede ameliorate asthma by inhibiting the crosstalk between ferroptosis and inflammation in airway epithelial cells.

Objectives: We previously revealed that scorpio and centipede (SC) improve the inflammatory response in asthma, whereas it is unclear whether ferroptosis is involved in this process. Materials and Methods: The asthmatic mouse model was established and lung tissues were collected for histopathological examination. The levels of tumor necrosis factor-α (TNF-α), interleukin- (IL-)1ß, Fe2+, malondialdehyde (MDA), glutathione peroxidase 4 (GPX4), ferritin heavy chain 1(FTH1), and reactive oxygen species (ROS) were assessed in asthmatic mice and mouse airway epithelial cells. Results: Our results showed that ferroptosis was induced in asthmatic mice, as evidenced by the reduction of FTH1 and GPX4 expression and the increase of MDA and Fe2+ levels (all P<0.05). Ferrostatin-1 repressed inflammation and ferroptosis of asthmatic mice. Additionally, SC significantly inhibited the levels of TNF-α, IL-1ß, MDA, and Fe2+, while enhancing FTH1 and GPX4 expression. However, SC plus erastin showed the reverse results. Moreover, ferroptosis remarkably increased in asthmatic airway epithelial cells, while SC suppressed ferroptosis of the cells (all P<0.05). Conclusion: SC ameliorated asthma by inhibiting the crosstalk between ferroptosis and inflammation in airway epithelial cells.

A Model for Predicting the Duration of Viral Shedding in Patients Who Had Been Hospitalized with Mild COVID-19: A Single-Center Retrospective Study.

Background: Clinical decision-making is enhanced by the development of a mathematical model for prognosis prediction. Screening criteria associated with viral shedding time and developing a prediction model facilitate clinical decision-making and are, thus, of great medical value. Methods: This study comprised 631 patients who were hospitalized with mild COVID-19 from a single center and 30 independent variables included. The data set was randomly divided into the training set (80%) and the validation set (20%). The outcome variable included viral shedding time and whether the viral shedding time >14 days, LASSO was used to screen the influencing factors. Results: There were 321 males and 310 females among the 631 cases, with an average age of 62.1 years; the median viral shedding time was 12 days, and 68.8% of patients experienced viral shedding within 14 days, with fever (50.9%) and cough (44.2%) being the most common clinical manifestations. Using LASSO with viral shedding time as the outcome variable, the model with lambda as 0.1592 (λ = 0.1592) and 13 variables (eg the time from diagnosis to admission, constipation, cough, hs-CRP, IL-8, IL-1ß, etc.) was more accurate. Factors were screened by LASSO and multivariable logistic regression with whether the viral shedding time >14 days as the outcome variable, five variables, including the time from diagnosis to admission, CD4 cell count, Ct value of ORF1ab, constipation, and IL-8, were included, and a nomogram was drawn; after model validation, the consistency index was 0.888, the AUC was 0.847, the sensitivity was 0.744, and the specificity was 0.830. Conclusion: A clinical model developed after LASSO regression was used to identify the factors that influence the viral shedding time. The predicted performance of the model was good, and it was useful for the allocation of medical resources.

Elucidation of the mechanisms and molecular targets of KeChuanLiuWei-Mixture for treatment of severe asthma based on network pharmacology.

KeChuanLiuWei-Mixture (KCLW) is widely used as a Chinese medicine prescription to treat severe asthma. However, the underlying therapeutic mechanism of KCLW remains unclear. In this study, a network pharmacology method was used to identify the chemical constituents of KCLW by the TCMSP database and ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry. Differential expression identification, protein-protein interaction (PPI) network and functional enrichment analysis were used to screen key targets of KCLW for severe asthma. Our results confirmed that quercetin, luteolin, kaempferol, and wogonin are the most critical active ingredients in KCLW. Moreover, the 16 relevant severe asthma-related targets of KCLW were obtained by overlapping the PPI networks of the KCLW putative targets and severe asthma-related genes, among which the most important targets were IL-6, NOS2, VEGFA, CXCL2, and PLAT. Functionally, the 16-targets and their interacting differentially expressed genes were primarily related to biological functions and pathways related to immunity and inflammation, such as inflammatory response, T cell differentiation, Nrf2/HO-1 signaling pathway, TGF-ß/Smad signaling pathway, and NF-κB signaling pathway. KCLW inhibited inflammation in PDGF-BB-induced airway smooth muscle cells. In summary, this study demonstrates the active substance and potential therapeutic mechanism of KCLW in severe asthma, and offers a clinical direction for KCLW against severe asthma.

Scorpion and centipede alleviates severe asthma through M2 macrophage-derived exosomal miR-30b-5p.

Asthma is one of the most common chronic inflammatory diseases. Although the scorpion and centipede (SC) significantly ameliorates asthma and changes exosomal miRNAs, the molecular mechanism is still obscure. Here, we show that SC improves inflammation in asthmatic mice and increases M2 macrophage-derived exosomes (M2Φ-Exos) by promoting M2 macrophage polarization. The M2Φ-Exos remarkably inhibits airway epithelial cell pyroptosis by reducing the expression of NLRP3, caspase-1, and LI-1ß and mitochondrial swelling. Furthermore, miR-30b-5p is up-regulated in M2Φ-Exos compared with M1Φ-Exos. Overexpression of miR-30b-5p in M2Φ-Exos prevents airway epithelial cell pyroptosis, while down-regulation of miR-30b-5p promotes pyroptosis. We also uncover that pyroptosis is increased in asthmatic mice, while SC blocks pyroptosis. Moreover, miR-30b-5p overexpressed M2Φ-Exos further enhances the ameliorative effect of SC, which significantly down-regulates IRF7 expression. Our results collectively reveal that M2Φ-Exos induced by SC could carry miR-30b-5p to mitigate severe asthma by inhibiting airway epithelial cell pyroptosis. Most importantly, our findings may provide a potential clinical application of M2Φ-Exos for treating severe asthma.

Small RNA Sequencing Reveals Exosomal miRNAs Involved in the Treatment of Asthma by Scorpio and Centipede.

Asthma is a common respiratory disease with inflammation in the lungs. Exosomes and microRNAs (miRNAs) play crucial role in inflammation, whereas the role of exosomal miRNA in asthma remains unknown. Here, we aimed to identify the key exosomal miRNAs and their underlying mechanisms involved in scorpio and centipede (SC) treatment in asthma. Eighteen mice were randomly divided into three groups: control group, asthma group, and SC treatment group. Effect of SC was assessed by hematoxylin-eosin staining and real-time PCR. Exosomes from asthma and SC treatment groups were analyzed by small RNA-seq. Results revealed SC significantly alleviated the pathogenesis of asthma and suppressed the release of inflammatory cytokines. A total of 328 exosomal miRNAs were differentially expressed between the exosomes from asthma and SC-treated mice, including 118 up- and 210 downregulated in SC-treated mice. The altered exosomal miRNAs were primarily involved in the function of transcription, apoptotic process, and cell adhesion; and pathway of calcium, Wnt, and MAPK signaling. Real-time PCR verified exosomal miR-147 was downregulated, while miR-98-5p and miR-10a-5p were upregulated in SC-treated mice compared to asthma mice. Moreover, the target genes of miR-147-3p, miR-98-5p, and miR-10a-5p were mainly enriched in Wnt and MAPK inflammatory signaling. miR-10a-5p promoted the proliferation of mouse lung epithelial cells and downregulated the expression of Nfat5 and Map2k6. These data suggest SC-induced exosomal miRNAs might mediate the inflammatory signaling and might be involved in the SC treatment in asthma. The exosomal miRNAs might be promising candidates for the treatment of asthma.

α-Asarone suppresses the proliferation and migration of ASMCs through targeting the lncRNA-PVT1/miR-203a/E2F3 signal pathway in RSV-infected rats.

Asthma is a chronic inflammatory pulmonary disease and respiratory syncytial virus (RSV) infection is a common cause of lower respiratory tract illness in infants and young children. α-Asarone presents many pharmacological effects and has been demonstrated to be useful in treating asthma. However, the functional mechanism of α-asarone in RSV-infected asthma has not been investigated. Long non-coding RNAs (lncRNAs) have been reported to play critical roles in many biological processes. Although many lncRNAs have been characterized, few were reported in asthma, especially in RSV-induced asthma. Currently, a novel post-transcriptional regulation has been proposed in which lncRNAs function as competing endogenous RNAs (ceRNAs) to competitively sponge miRNAs, thereby regulating the target genes. In the present study, we established an RSV-infected Sprague-Dawley rat model and demonstrated that lncRNA-PVT1 is involved in the mechanism of α-asarone in treating RSV-induced asthma, and lncRNA-PVT1 regulates the expression of E2F3 by functioning as a ceRNA which competitively sponges miR-203a.

Length-dependent ß-sheet growth mechanisms of polyalanine peptides in water and on hydrophobic surfaces.

Fibrillar assemblies by peptides are becoming one of the most promising nanomaterials due to their exceptional properties. The self-assembly of peptides into ß sheets is a critical step in the fibrillization pathway. We investigated the length-dependent ß-sheet growth mechanisms of polyalanine [poly(A)] peptides consisting of 6 to 24 alanines (A6 to A24) in water and on the hydrophobic surface, respectively, by molecular dynamics simulations. ß-sheet growth behavior in water fits negative exponential growth model, showing that ß-sheet growth rate decays exponentially with time. Meanwhile, increasing chain length leads to an accelerated decay of the ß-sheet growth rate. By contrast, ß-sheet growth on the surface from A6 to A18 occurs in two consecutive stages, both of which fit linear growth models. ß-sheet growth rate in the first stage increases as chain length is increased, while the intermediate length peptide A12 has the highest ß-sheet growth rate in the second stage. ß-sheet growth behavior of A24 on the surface still fits negative exponential model. Overall, the hydrophobic surface accelerates ß-sheet growth by enhancing local concentration and reducing conformational entropy of poly(A) peptide, and the ß-sheet growth of the intermediate length peptide A12 is the fastest on the surface. Our simulation results shed light on understanding the accelerated peptide fibrillization on the hydrophobic surface.

Effect of "yang-warming and kidney essence-replenishing" herbal paste on cold-related asthma exacerbation.

OBJECTIVE: To observe the effect of "Yang-warming and kidney essence-replenishing" herbal paste on cold-related asthma exacerbation. METHODS: One hundred and fifty one patients with moderate to severe persistent asthma were randomly divided into a treatment group (n = 74) and a control group (n = 69). Both groups were given basic treatment according to the Global Initiative for Asthma scheme. The treatment group was also treated with herbal paste in the winter. The frequency of asthma exacerbation, cold and cold-related asthma exacerbation, scores of the asthma control test (ACT), and kidney-deficiency syndrome during the one-year follow-up in each group were recorded. RESULTS: Compared with the control group, the onset frequencies of catching cold, asthma exacerbation, cold-related asthma exacerbation, and kidney-deficiency syndrome score were significantly reduced. There was no significant difference in scores of ACT between the two groups. CONCLUSION: "Yang-warming and kidney essence-replenishing" herbal paste could reduce cold-related asthma exacerbation.

[Phenotype change of airway smooth muscle cell in bronchial asthma rats and the intervention of Zhichuan capsule on it].

OBJECTIVE: To investigate the phenotype change of airway smooth muscle cell (SMC) in bronchial asthma and the effect of Zhichuan Capsule (ZCC) on it. METHODS: Sixty male SD rats were randomized into 5 groups equally, the normal control group, the model group, and three treated groups treated with ZCC, Western medicine (WM) and ZCC + WM respectively. Excepting rats in the control group, they were all made into bronchial asthma model rats and received respective treatment. Histomorphology of the airway SMC was observed by electron microsope at the 4th week, and the expressions of smalpha-actin and sm-MHC were examined at the 2nd and the 4th week after provocation. RESULTS: As compared with the control group, moyfilaments in SMC were lesser, secretive organelles were more and levels of smalpha-actin and sm-MHC expressions at the 4th week were lower in the model group (P < 0.05). While histomorphology of SMC in the WM group was as narmal, but some hypoxia changes were seen, such as the fracture of mitochondria cristae and deplation of matrix; histomorphology of SMC in the ZCC treated group and the ZCC + WM treated group were similar to that in the normal control group, and their expressions of smalpha-actin and sm-MHC were enhanced as compared to those in the model group (P < 0.05). CONCLUSION: Airway SMC changes from contractile phenotype to synthetic phenotype in asthma rats, but the progress of change may be inhibited by ZCC.