[Network pharmacological analysis and experimental study of Pulsatilla chinensis against inflammatory injury caused by pneumonia in mice infected with influenza virus FM_1].

Network pharmacology and the mouse model of viral pneumonia caused by influenza virus FM_1 were employed to explore the main active components and the mechanism of Pulsatilla chinensis against the inflammatory injury of influenza virus-induced pneumonia. The components and targets of P. chinensis were searched from TCMSP, and the targets associated with influenza virus-induced pneumonia were searched from GeneCards. The common targets between P. chinensis and influenza virus-induced pneumonia were identified with Venn diagram established in Venny 2.1. The herb-component-disease-target(H-C-D-T) network was constructed by Cytoscape 3.7.2. The above data were imported into STRING for PPI network analysis. Gene Ontology(GO) enrichment and KEGG pathway enrichment were performed with DAVID. BALB/cAnN mice were infected with the influenza virus FM_1 by nasal drip to gene-rate the mouse model of pneumonia. Immunohistochemistry was adopted to the expression profiling of inflammatory cytokines in the lung tissues of mice in the blank group, model group, and P. chinensis group 1, 3, 5, and 7 days after infection. The pathological changes of lung and trachea of mice in blank group, model group, and P. chinensis group were observed with light microscope and scanning electron microscope at all the time points. The network pharmacological analysis indicated that 9 compounds of P. chinensis were screened out, with a total of 57 targets, 22 of which were overlapped with those of influenza virus-induced pneumonia. A total of 112 GO terms(P<0.05) were enriched, including 81 terms of biological processes, 11 terms of cell components, and 20 terms of molecular functions. A total of 53 KEGG signaling pathways(P<0.05) were enriched, including TNF signaling pathway, influenza A signaling pathway, NF-κB signaling pathway, MAPK signaling pathway and other signaling pathways related to influenza/inflammation. In the P. chinensis group, the expression of TNF-α and IL-1 in the lung tissue was down-regulated on the 3 rd day after infection, and that of IL-6 in the lung tissue was down-regulated on the 5 th day after infection. Light microscopy and scanning electron microscopy showed that P. chinensis significantly alleviated the pathological damage of lung and trachea compared with the model group. This study reflects the multi-components, multi-targets, and multi-pathways of P. chinensis against influenza virus-induced pneumonia. P. chinensis may reduce the production of proinflammatory cytokines and mediators and block the pro-inflammatory signaling pathways to alleviate viral pneumonia, which provides reference for future research.

The characteristics of the frequent exacerbator with chronic bronchitis phenotype and non-exacerbator phenotype in patients with chronic obstructive pulmonary disease: a meta-analysis and system review.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) patients with different phenotypes show different clinical characteristics. Therefore, we conducted a meta-analysis to explore the clinical characteristics between the non-exacerbator (NE) phenotype and the frequent exacerbator with chronic bronchitis (FE-CB) phenotype among patients with COPD. METHODS: CNKI, Wan fang, Chongqing VIP, China Biology Medicine disc, PubMed, Cochrane Library, and EMBASE databases were searched from the times of their inception to April 30, 2019. All studies that reported the clinical characteristics of the COPD phenotypes and which met the inclusion criteria were included. The quality assessment was analyzed by Cross-Sectional/Prevalence Study Quality recommendations. The meta-analysis was carried out using RevMan5.3. RESULTS: Ten cross-sectional observation studies (n = 8848) were included. Compared with the NE phenotype, patients with the FE-CB phenotype showed significantly lower forced expiratory volume in 1 s percent predicted (FEV1%pred) (mean difference (MD) -8.50, 95% CI -11.36--5.65, P < 0.001, I2 = 91%), forced vital capacity percent predicted (FVC%pred) [MD - 6.69, 95% confidence interval (CI) -7.73--5.65, P < 0.001, I2 = 5%], and forced expiratory volume in 1 s/forced vital capacity (FEV1/FVC) (MD -3.76, 95% CI -4.58--2.95,P < 0.001, I2 = 0%); in contrast, Charlson comorbidity index (MD 0.47, 95% CI 0.37-0.58, P < 0.001, I2 = 0], COPD assessment test (CAT) score (MD 5.61, 95% CI 4.62-6.60, P < 0.001, I2 = 80%), the quantity of cigarettes smoked (pack-years) (MD 3.09, 95% CI 1.60-4.58, P < 0.001, I2 = 41%), exacerbations in previous year (2.65, 95% CI 2.32-2.97, P < 0.001, I2 = 91%), modified Medical British Research Council (mMRC) score (MD 0.72, 95% CI 0.63-0.82, P < 0.001, I2 = 57%), and body mass index (BMI), obstruction, dyspnea, exacerbations (BODEx) (MD 1.78, 95% CI 1.28-2.28, P < 0.001, I2 = 91%), I2 = 34%) were significantly higher in patients with FE-CB phenotype. No significant between-group difference was observed with respect to BMI (MD-0.14, 95% CI -0.70-0.42, P = 0.62, I2 = 75%). CONCLUSION: COPD patients with the FE-CB phenotype had worse pulmonary function and higher CAT score, mMRC scores, frequency of acute exacerbations, and the quantity of cigarettes smoked (pack-years) than those with the NE phenotype.

Effect of acupoint application on T lymphocyte subsets in patients with chronic obstructive pulmonary disease: A meta-analysis.

BACKGROUND: The development of chronic obstructive pulmonary disease (COPD) is related to the T lymphocyte mediated inflammatory immune response and immune imbalance. The purpose of this systematic review was to evaluate the clinical efficacy and safety of acupoint application on T lymphocyte subsets in patients with COPD. METHODS: We searched CNKI, Wan fang, Chongqing VIP, China Biology Medicine disc, PubMed, the Cochrane Library, and EMBASE for studies published as of Oct. 31, 2019. All randomized controlled trials of acupoint application on COPD patients that met the inclusion criteria were included. The Cochrane bias risk assessment tool was used for literature evaluation. RevMan5.3 software was used for meta-analysis. RESULTS: Eight studies (combined n = 524) qualified based on the inclusion criteria. Compared with routine treatment alone, acupoint application combined with routine treatment can significantly increase the T lymphocyte CD4/CD8 ratio (MD 0.12, 95% CI 0.03-0.21, P < .01, I = 49%), reduce CD8 T-cells (MD-0.99, 95% CI-1.70-0.28, P < .001, I = 37%), reduce the times of acute exacerbations (MD-0.28, 95% CI-0.35-0.21, P < .001, I = 0), and improve the clinical efficacy (MD 1.30, 95% CI 1.14-1.48, P < .001, I = 39%). CONCLUSION: Acupoint application can improve the CD4/CD8 ratio and CD8 T-cells in patients with COPD and has an auxiliary effect in reducing the times of acute exacerbations and improving clinical efficacy.

The characteristics of the frequent exacerbators with chronic bronchitis phenotype and the asthma-chronic obstructive pulmonary disease overlap syndrome phenotype in chronic obstructive pulmonary disease patients: A meta-analysis and system review.

To investigate the difference of clinical characteristics between chronic obstructive pulmonary disease (COPD) patients with the frequent exacerbators with chronic bronchitis (FE-CB) phenotype and those with the asthma-COPD overlap syndrome (ACO) phenotype.We searched CNKI, Wan Fang, Chongqing VIP, China Biology Medicine disc, PubMed, Cochrane Library, and EMBASE databases for studies published as of April 30, 2019. All studies that investigated COPD patients with the FE-CB and ACO phenotypes and which qualified the inclusion criteria were included. Cross-sectional/prevalence study quality recommendations were used to measure methodological quality. RevMan5.3 software was used for meta-analysis.Ten studies (combined n = 4568) qualified the inclusion criteria. The FE-CB phenotype of COPD was associated with significantly lower forced vital capacity percent predicted (mean difference [MD] -9.05, 95% confidence interval [CI] [-12.00, -6.10], P < .001, I = 66%), forced expiratory volume in 1 second (FEV1) (MD -407.18, 95% CI [-438.63, -375.72], P < .001, I = 33%), forced expiratory volume in 1 second percent predicted (MD -9.71, 95% CI [-12.79, -6.63], P < .001, I = 87%), FEV1/forced vital capacity (MD -5.4, 95% CI [-6.49, -4.30], P < .001, I = 0%), and body mass index (BMI) (MD -0.81, 95% CI [-1.18, -0.45], P < .001, I = 44%) as compared to the ACO phenotype. However, FE-CB phenotype was associated with higher quantity of cigarettes smoked (pack-years) (MD 6.45, 95% CI [1.82, 11.09], P < .001, I = 73%), COPD assessment test score (CAT) (MD 4.04, 95% CI [3.46, 4.61], P < .001, I = 0%), mMRC score (MD 0.54, 95% CI [0.46, 0.62], P < .001, I = 34%), exacerbations in previous year (1.34, 95% CI [0.98, 1.71], P < .001, I = 68%), and BMI, obstruction, dyspnea, exacerbations (BODEx) (MD 1.59, 95% CI [1.00, 2.18], P < .001, I = 86%) as compared to the ACO phenotype.Compared with the ACO phenotype, COPD patients with the FE-CB phenotype had poorer pulmonary function, lower BMI, and higher CAT score, quantity of cigarettes smoked (pack-years), exacerbations in previous year, mMRC score, and BODEx.This study is an analysis of published literature, which belongs to the second study. Therefore, this study does not require the approval of the ethics committee. The findings will be disseminated through a peer-reviewed journal publication or conference presentation.

[Effect of Scutellariae Radix on expression of inflammatory cytokine protein and gene in lung of mice with viral pneumonia caused by influenza virus FM1 infection].

Mice models of viral pneumonia were induced by pulmonary adaptive strain FM1 of influenza A virus in Asian mice.RT-PCR and immunohistochemistry were used to dynamically observe the effect of Scutellariae Radix on the protein and gene expression of inflammatory cytokine in the lungs of the model mice infected by influenza virus FM1 at different phases. The partial mechanism of Scutellariae Radix in repairing the immune inflammatory damage of target organs of pneumonia caused by influenza virus was further explored. The results showed that Scutellariae Radix reduced protein and gene expression of proinflammatory cytokines tumor necrosis factor( TNF-α),interleukin IL-1,IL-6 in lung tissues from 3 rd to 5 th day after infection,and increased protein and gene expression of IL-10 and IFN-γ in lung tissues on the 5 th day after infection. Scutellariae Radix may inhibit excessive release of pro-inflammatory cytokines and promote the expression of anti-inflammatory cytokines,thereby inhibiting the systemic inflammatory response syndrome,reducing the immunoinflammatory pathological damage of lung caused by influenza virus FM1 infection,and promoting lung repair of tissue inflammatory lesions.

[Mechanism of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture against influenza immune inflammatory injury].

OBJECTIVE: To observe the impact of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture on mRNA expression of lung inflammatory cytokines and pulmonary pathological injury of mice infected by influenza virus, in order to discuss the mechanism of tonifying Qi traditional Chinese medicines against pulmonary immune inflammatory injury of infected mice. METHOD: In different time phases after mice were infected with influenza virus FM1, the RT-PCR method was adopted to observe the impact of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture on five inflammatory cytokines TNF-α, IL-1, IL-6, IL-10 and IFN-γ, and the changes in pulmonary pathological injury of mice with viral pneumonia after intervention with tonifying qi traditional Chinese medicines. RESULT: (1) Tonifying Qi traditional Chinese medicines significantly reduced the mRNA expression of TNF-α at 1-5 d and IL-1 mRNA expression at 7 d, may increase IL-1 mRNA expression in mouse lung at 3 d, significantly reduced IL-6 mRNA expression in mouse lung and increased IL-10 mRNA expression at 3-7 d, and significantly increased IFN-γ mRNA expression at 1 d. (2) Tonifying Qi traditional Chinese medicines could significantly inhibited and repaired pulmonary immune inflammatory injury of mice infected by FM1, which was most remarkable at 3-7 d after the infection with influenza virus FM1. CONCLUSION: Tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture could resist pulmonary immune inflammatory injury and repair inflammatory injury by regulating the mRNA expression of imbalance inflammatory cytokines of organisms infected with influenza virus.

[The influence of representative herbs of clearing and detoxifying drugs effect on inflammatory cytokines expression of mice lung homogenate infected by influenza virus FM1].

OBJECTIVE: In order to screen out a certain kind of traditional medicine which has a better role in immune regulatory, the influence of representatives of heat clearing and detoxicating herb on inflammatory cytokines protein expression of mice lung homogenate infected by FM1 have been observed. METHOD: Modeling mice infected by FM1. On the first, third, fifth and seventh day after FM1 infection, tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-10 (IL-10), and gamma-interferon (IFN-gamma) expression in mice lung homogenate of normal control group, model control group, scutellari group, isatidis group, pulsatilla group, polygonum cuspidatum group and oldenlandia group have been tested by ELISA method. RESULT: The expression of TNF-alpha, IL-6, IFN-gamma and IL-10 in mice lung homogenate reaches its peak on the third day after FM1 infection, significantly higher than the control group (P < 0.05). Scutellari and isatidis are two representatives of heat clearing and detoxicating herb, which can decrease the expression of TNF-alpha, IL-6 and IL-1 and increase the expression of IL-10, IFN-gamma. The effect are more pronounced and statistically significant (P < 0.05) on the third and fifth day after infection, pulsatilla, polygonum cuspidatum and oldenlandia can also regulate the inflammatory cytokines, but the effect are not so obvious as scutellari and isatidis. CONCLUSION: Scutellari and isatidis, two representatives of heat clearing and detoxicating herb, have a good intervention on immune damage caused by influenza virus through adjusting the balance of inflammatory cytokines and anti-inflammatory cytokines.

[Study on the effect of realgar nanoparticles anti-adenovirus in vitro].

OBJECTIVE: This study was to establish a model that adenovirus type 3 (HAdV-3) infected on Hep-2 cell in order to explore anti-adenovirus3 (HAdV-3) effect of Chinese medicine realgar in vitro. METHOD: Use high-energy ball milling with distilled water to prepare realgar nanoparticles. The concentration of nanometer realgar was tested by molybdenum blue staining method and realgar nanoparticles' particle size was tested on Nano Series. The technique of cell culture with ribavirin as positiv control was to observe anti-adenovirus effect through prevention, treatment and direct inactivation of three kinds of drug delivery. RESULT: This drug was found to be a potential inhibitor of HAdV-3 in a concentration-dependent manner with the median toxic concentration (TC50) of 0.649 microg/ml in Hep-2 Cell culture. The median inhibition concentration (IC50) was 0.255 microg/ml when drug was added before infection. The IC50 was 0.142 microg/ml when drug was added after virus infection, and it was 0.117 microg/ml as the drug was added after it mixed with virus. The therapeutic index (TI) was 2.55, 4.57 and 5.55 respectively. CONCLUSION: The direct inactivation effect of realgar nanoparticles is the most evident in three drug deliveries manner with the same concentration in vitro.

[The dynamic effect of yiqi qingwen jiedu heji on the protein expressions of cytokine IFN-gamma, TNF-alpha, IL-10 and IL-6 in the lungs of mice infected by IV FM1].

OBJECTIVE: To find the function and functioning mechanism of Yiqi Qingwen Jiedu Heji in resisting influenza immune damage by studying its effect on the expressions of cytokine. METHOD: Taking IV FM1 infected mice as its model and doing ELISA (double antibody sandwichenzyme linked immunosorbent assay), we dynamically observed the change of cytokine TNF-alpha, IL-6, IFN-gamma and IL-10 after giving Yiqi Qingwen Jiedu Heji treatment. RESULT AND CONCLUSION: After the mice are infected by influenza virus, their protein expressions of the model group are higher than those of the control group, of which TNF-alpha, IL-6, IFN-gamma reach the peak in three days. The three expressions of Yiqi Qingwen Jiedu Heji treated group are decreased and the decrease becomes remarkable on the third day, compared with those of the model group. However, the expression of IL-10 of the treated group is remarkably increased. It indicates that Yiqi Qingwen Jiedu Heji can resist the expressions of TNF-alpha, IL-6 and IFN-gamma pro-inflammatory cytokine,increase the expression of IL-10, and thus, alleviate inflammatory injury. So the clinical application of such medicine can shorten the course of disease.