Periplogenin inhibits pathologic synovial proliferation and infiltration in rheumatoid arthritis by regulating the JAK2/3-STAT3 pathway.

Rheumatoid arthritis (RA) is an autoimmune disease that affects joints, causing inflammation, synovitis, and erosion of cartilage and bone. Periplogenin is an active ingredient in the anti-rheumatic and anti-inflammatory herb, cortex periplocae. We conducted a study using a CIA model and an in vitro model of fibroblast-like synoviocytes (FLS) induced by Tumor Necrosis Factor-alpha (TNF-α) stimulation. We evaluated cell activity, proliferation, and migration using the CCK8 test, EDU kit, and transwell assays, as well as network pharmacokinetic analysis of periplogenin targets and RA-related effects. Furthermore, we measured inflammatory factors and matrix metalloproteinases (MMPs) expression using ELISA and qRT-PCR assays. We also evaluated joint destruction using HE and Safranin O-Fast Green Staining and examined the changes in the JAK2/3-STAT3 pathway using western blot. The results indicated that periplogenin can effectively inhibit the secretion of inflammatory factors, suppress the JAK2/3-STAT3 pathway, and impede the proliferation and migration of RA FLS. Thus, periplogenin alleviated the Synovial inflammatory infiltration of RA.

MetNC: Predicting Metabolites in vivo for Natural Compounds.

Natural compounds (NCs) undergo complicated biotransformation in vivo to produce diverse forms of metabolites dynamically, many of which are of high medicinal value. Predicting the profiles of chemical products may help to narrow down possible candidates, yet current computational methods for predicting biotransformation largely focus on synthetic compounds. Here, we proposed a method of MetNC, a tailor-made method for NC biotransformation prediction, after exploring the overall patterns of NC in vivo metabolism. Based on 850 pairs of the biotransformation dataset validated by comprehensive in vivo experiments with sourcing compounds from medicinal plants, MetNC was designed to produce a list of potential metabolites through simulating in vivo biotransformation and then prioritize true metabolites into the top list according to the functional groups in compound structures and steric hindrance around the reaction sites. Among the well-known peers of GLORYx and BioTransformer, MetNC gave the highest performance in both the metabolite coverage and the ability to short-list true products. More importantly, MetNC seemed to display an extra advantage in recommending the microbiota-transformed metabolites, suggesting its potential usefulness in the overall metabolism estimation. In summary, complemented to those techniques focusing on synthetic compounds, MetNC may help to fill the gap of natural compound metabolism and narrow down those products likely to be identified in vivo.

Different Smoking Statuses on Survival and Emphysema in Patients with Acute Exacerbation of Chronic Obstructive Pulmonary Disease.

Objective: To investigate the association between different smoking statuses and survival and emphysema in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Methods: This retrospective study included patients admitted from October 2014 to September 2017. Demographic, clinical, laboratory, imaging, impulse oscillometry, and traditional pulmonary function data were collected. The relationship between smoking and EI was analyzed via binary logistic regression after adjusting for other factors. Survival was analyzed using the Kaplan-Meier method and the log rank test. Results: The patients with AECOPD (357 cases) were identified (and stratified into three groups: never smoked (NS; n=83), former smokers (FS, n=118), and current smokers (CS; n=156). Compared with CS, NS were older and predominantly female. No differences were observed in respiratory symptoms and acute exacerbation between CS and NS. NS had higher resistance and reaction in the central and peripheral airways, while CS exhibited more severe diffuse dysfunction. CS demonstrated more severe and extensive emphysema. Smoking was an independent risk factor for emphysema after adjusting for age, forced expiratory volume in the first second over predicted value, BMI, leukocyte count, and carbon monoxide transfer coefficient. No difference in 5-year survival rates between NS and CS was established. Conclusion: CS has the worst pulmonary function, suggesting a more important destruction of the lung parenchyma, while AECOPD without smoking risk factors mostly affects the airways. Impulse oscillometry can be used for imaging airway-dominant AECOPD. There was no difference in the 5-year survival rate.

Potential Value of Expiratory CT in Quantitative Assessment of Pulmonary Vessels in COPD.

Objective: To investigate the associations between intrapulmonary vascular volume (IPVV) depicted on inspiratory and expiratory CT scans and disease severity in COPD patients, and to determine which CT parameters can be used to predict IPVV. Methods: We retrospectively collected 89 CT examinations acquired on COPD patients from an available database. All subjects underwent both inspiratory and expiratory CT scans. We quantified the IPVV, airway wall thickness (WT), the percentage of the airway wall area (WA%), and the extent of emphysema (LAA%-950) using an available pulmonary image analysis tool. The underlying relationship between IPVV and COPD severity, which was defined as mild COPD (GOLD stage I and II) and severe COPD (GOLD stage III and IV), was analyzed using the Student's t-test (or Mann-Whitney U-test). The correlations of IPVV with pulmonary function tests (PFTs), LAA%-950, and airway parameters for the third to sixth generation bronchus were analyzed using the Pearson or Spearman's rank correlation coefficients and multiple stepwise regression. Results: In the subgroup with only inspiratory examinations, the correlation coefficients between IPVV and PFT measures were -0.215 ~ -0.292 (p < 0.05), the correlation coefficients between IPVV and WT3-6 were 0.233 ~ 0.557 (p < 0.05), and the correlation coefficient between IPVV and LAA%-950 were 0.238 ~ 0.409 (p < 0.05). In the subgroup with only expiratory scan, the correlation coefficients between IPVV and PFT measures were -0.238 ~ -0.360 (p < 0.05), the correlation coefficients between IPVV and WT3-6 were 0.260 ~ 0.566 (p < 0.05), and the correlation coefficient between IPVV and LAA%-950 were 0.241 ~ 0.362 (p < 0.05). The multiple stepwise regression analyses demonstrated that WT were independently associated with IPVV (P < 0.05). Conclusion: The expiratory CT scans can provide a more accurate assessment of COPD than the inspiratory CT scans, and the airway wall thickness maybe an independent predictor of pulmonary vascular alteration in patients with COPD.

Role of the Emphysema Index Combined with the Chronic Obstructive Pulmonary Disease Assessment Test Score in the Evaluation of Chronic Obstructive Pulmonary Disease.

Background: This study aimed to evaluate the efficacy of the emphysema index (EI) in distinguishing chronic bronchitis (CB) from chronic obstructive pulmonary disease (COPD) and its role, combined with the COPD Assessment Test (CAT) score, in the evaluation of COPD. Methods: A total of 92 patients with CB and 277 patients with COPD were enrolled in this study. Receiver operating characteristic (ROC) curves were analyzed to evaluate whether the EI can preliminarily distinguish chronic bronchitis from COPD. Considering the heterogeneity of COPD, there might be missed diagnosis of some patients with bronchitis type when differentiating COPD patients only by EI. Therefore, patients with COPD were classified according to the CAT score and EI into four groups: Group 1 (EI < 16%, CAT < 10), Group 2 (EI < 16%, CAT ≥ 10), Group 3 (EI ≥ 16%, CAT < 10), and Group 4 (EI ≥ 16%, CAT ≥ 10). The records of pulmonary function and quantitative computed tomography findings were retrospectively analyzed. Results: ROC curve analysis showed that EI = 16.2% was the cutoff value for distinguishing COPD from CB. Groups 1 and 2 exhibited significantly higher maximal voluntary ventilation (MVV) percent predicted (pred), forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC), maximal midexpiratory flow of 25-75% pred, carbon monoxide-diffusing capacity (DLCO)/alveolar ventilation (VA), FEV1 % pred (p ≤ 0.013), and maximal expiratory flow 50% pred (all p < 0.05) than Group 4. FEV1/FVC and DLCO/VA were significantly lower in Group 3 than in Group 2 (p=0.002 and p < 0.001, respectively). The residual volume/total lung capacity was higher in Group 3 than in Groups 1 and 2 (p < 0.05). Conclusions: The combination of EI and CAT was effective in the evaluation of COPD.

Nitidine chloride inhibits fibroblast like synoviocytes-mediated rheumatoid synovial inflammation and joint destruction by targeting KCNH1.

OBJECTIVE: Nitidine chloride (NC), a natural small molecular compound from traditional Chinese herbal medicine zanthoxylum nitidum, has been shown to exhibit anti-tumor effect. However, its role in autoimmune diseases such as rheumatoid arthritis (RA) is unknown. Here, we investigate the effect of NC in controlling fibroblast-like synoviocytes (FLS)-mediated synovial inflammation and joint destruction in RA and further explore its underlying mechanism(s). METHODS: FLSs were separated from synovial tissues obtained from patients with RA. Protein expression was analyzed by Western blot or immunohistochemistry. Gene expression was measured using quantitative RT-PCR. ELISA was used to measure the levels of cytokines and MMPs. Cell proliferation was detected using EdU incorporation. Migration and invasion were evaluated by Boyden chamber assay. RNA sequencing analysis was used to identify the target of NC. Collagen-induced arthritis (CIA) model was used to evaluate the in vivo effect of NC. RESULTS: NC treatment reduced the proliferation, migration, invasion, and lamellipodia formation but not apoptosis of RA FLSs. We also demonstrated the inhibitory effect of NC on TNF-α-induced expression and secretion of IL-6, IL-8, CCL-2, MMP-1 and MMP-13. Furthermore, we identified KCNH1, a gene that encodes ether-à-go-go-1 channel, as a novel targeting gene of NC in RA FLSs. KCNH1 expression was increased in FLSs and synovial tissues from patients with RA compared to healthy controls. KCNH1 knockdown or NC treatment decreased the TNF-α-induced phosphorylation of AKT. Interestingly, NC treatment ameliorated the severity of arthritis and reduced synovial KCNH1 expression in mice with CIA. CONCLUSIONS: Our data demonstrate that NC treatment inhibits aggressive and inflammatory actions of RA FLSs by targeting KCNH1 and sequential inhibition of AKT phosphorylation. Our findings suggest that NC might control FLS-mediated rheumatoid synovial inflammation and joint destruction, and be a novel therapeutic agent for RA.

ß-Sitosterol Inhibits Rheumatoid Synovial Angiogenesis Through Suppressing VEGF Signaling Pathway.

Background: Rheumatoid arthritis (RA) is a chronic disabling inflammatory disease that causes synovial angiogenesis in an invasive manner and leads to joint destruction. Currently available pharmacotherapy for RA has unwanted side effects and limitations. Although anti-angiogenic therapy is regarded as a new potential treatment for RA, only a few anti-angiogenic drugs are available. An increasing number of studies have shown that ß-sitosterol (BSS) may exert inhibitory effects against angiogenesis. However, the mechanisms involved are still unclear. Methods: Based on the results of the gene set enrichment analysis (GSEA) of the transcriptome data of endothelial cells from RA patients, we evaluated the pharmacological effects of BSS on the tube formation, cell proliferation, and migration of human umbilical vein endothelial cells (HUVECs). Furthermore, the effects of BSS treatment on vascular endothelial growth factor receptor 2 (VEGFR2) were determined using molecular docking and Western blotting. Additionally, in the presence or absence of BSS, synovial angiogenesis and joint destruction of the ankle were investigated in collagen-induced arthritis (CIA) mice. The effect of BSS treatment on VEGFR2/p-VEGFR2 expression was verified through immunohistochemical staining. Results: The immunohistochemistry results revealed that BSS treatment inhibited angiogenesis both in vitro and in vivo. In addition, the results of 5-ethynyl-2'-deoxyuridine and cell cycle analysis showed that BSS treatment suppressed the proliferation of HUVECs, while the Transwell migration and stress fiber assays demonstrated that BSS treatment inhibited the migration of HUVECs. Notably, the inhibitory effect of BSS treatment on VEGFR2/p-VEGFR2 was similar to that of axitinib. In CIA mice, BSS also exerted therapeutic effects on the ankles by reducing the degree of swelling, ameliorating bone and cartilage damage, preventing synovial angiogenesis, and inhibiting VEGFR2 and p-VEGFR2 expression. Conclusion: Therefore, our findings demonstrate that BSS exerts an inhibitory effect on synovial angiogenesis by suppressing the proliferation and migration of endothelial cells, thereby alleviating joint swelling and bone destruction in CIA mice. Furthermore, the underlying therapeutic mechanisms may involve the inhibition of VEGF signaling pathway activation.

Imaging Features of Chronic Bronchitis with Preserved Ratio and Impaired Spirometry (PRISm).

PURPOSE: The purpose of the study was to investigate the quantitative chest tomographic features of chronic bronchitis with preserved ratio and impaired spirometry (PRISm), including airway wall area, emphysema index, and lung capacity. METHODS: An observational, cross-sectional study of 343 patients at the Ninth Hospital of Xi'an Affiliated Hospital of Xi'an Jiaotong University between October 2014 and September 2017. The patients were divided into three groups: 77 cases of chronic bronchitis with normal lung function (forced expiratory volume in one second/forced vital capacity) (FEV1/FVC > 70%, FEV1%pred > 80%), 80 cases of chronic bronchitis with PRISm (FEV1/FVC > 70%, FEV1%pred < 80%), and 186 cases of the early chronic obstructive pulmonary disease (COPD) (FEV1/FVC < 70%, FEV1%pred > 50%, that is, Global Initiative for Chronic Obstructive Lung Disease (GOLD) grade 1 + 2). We compared and analyzed the differences in imaging between the chronic bronchitis with PRISm and the other two groups. RESULTS: Compared with the early COPD group, the PRISm group revealed significant differences in airway wall area, emphysema index, and lung capacity (P < 0.05). Compared with the chronic bronchitis with normal lung function group, the PRISm group showed increased WA%LUL5, decreased lung capacity, and higher mean lung density. CONCLUSION: In terms of airway wall area and emphysema index, patients with chronic bronchitis with PRISm were essentially no different than those with chronic bronchitis without abnormal spirometry, whereas for symptoms, they are more like GOLD 1 and 2 patients. Our findings show that it is not yet clear whether it constitutes an intermediate stage of chronic bronchitis with normal lung function that progression to early COPD.

Impulse oscillometry system as an alternative diagnostic method for chronic obstructive pulmonary disease.

We aimed to compare impulse oscillation system (IOS) and traditional pulmonary function tests (PFTs) for the assessment of the severity of chronic obstructive pulmonary disease (COPD), and to assess the use of IOS parameters to identify patients who were forced expiratory volume in 1 second (FEV1)%pred < 50%.Patients with COPD (n = 215) were enrolled at the Ninth Hospital of Xi'an Affiliated Hospital of Xi'an Jiaotong University between October 2014 and September 2016. All patients were assessed by traditional PFT and IOS. Diagnostic performance of IOS parameters to determine indication for patients of FEV1%pred < 50% was assessed on receiver-operating characteristics (ROC) curve analysis.Out of 215 patients, 18, 83, 78, and 36 patients were classified as grade 1, 2, 3, and 4, respectively, according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) severity grading. On Spearman correlation analysis, FEV1%pred, MMEF 75%-25%, and residual volume/total lung capacity (RV/TLC) correlated with total respiratory impedance (Z5)%pred, resistance at 5 Hz (R5)-resistance at 20 Hz (R20), R5-R20% R5, R5, R5%pred, frequency response (Fres), reactance area (Ax), and reactance at 5 Hz (X5). On ROC curve analysis, the area under the curve (AUC) of X5 absolute value, Fres, Ax, Z5%pred, R5-R20, and R5-R20% R5 were 0.748, 0.755, 0.760, 0.705, 0.715, and 0.735, respectively, for COPD patients who required inhalational glucocorticoid therapy.IOS parameters showed a good correlation with traditional pulmonary function parameters; reactance parameters showed a stronger correlation than that of the resistance parameters. IOS can be used as an alternative method for pulmonary function assessment in patients with COPD with FEV1%pred < 50% who need inhalational glucocorticoid therapy. CLINICAL TRIAL REGISTRATION NUMBER: ChiCTR-OCH-14004904.

[Astragalus polysaccharide alleviates lipopolysaccharide-induced lung injury in rats by inhibiting neutrophil activation].

Objective To study the effects of Astragalus polysaccharide (APS) on rat lung injury induced by lipopolysaccharide (LPS). Methods A total of 36 adult SD rats were randomly divided into control group, LPS model group and APS administration group. Acute lung injury was induced by a single intratracheal injection of LPS. Lung injury in rats was observed by HE staining. The changes of proinflammatory factors such as tumor necrosis factor α (TNF-α) and interleukin 1ß (IL-1ß) were detected by ELISA. The numbers of white blood cells (WBCs) and neutrophils in bronchoalveolar lavage fluid (BALF) were determined by cell counting. The activity of myeloperoxidase (MPO) and reactive oxygen species (ROS) were tested by the respective kits. Results Compared with the control group, the LPS-induced model group showed pulmonary edema and increased vascular permeability, upregulated levels of TNF-α and IL-1ß in BALF, significantly ascended numbers of WBCs and neutrophils, enhanced MPO activity and increased ROS level. However, administration of APS suppressed LPS-induced vascular permeability, tissue injury and inflammation in the rat lung. APS also suppressed LPS-induced neutrophilic inflammation in BALF. MPO activity and ROS level decreased significantly as well. Conclusion Administration of APS suppresses LPS-induced lung injury via inhibiting of neutrophil activation in rats.

miR-26a desensitizes non-small cell lung cancer cells to tyrosine kinase inhibitors by targeting PTPN13.

Epidermal growth factor receptor (EGFR)-targeted tyrosine kinase inhibitors (TKIs) have emerged as first-line drugs for non-small cell lung cancers (NSCLCs). However, the resistance to TKIs represents the key limitation for their therapeutic efficacy. We found that miR-26a was upregulated in gefitinib-refractory NSCLCs; miR-26a is downstream of EGFR signaling and directly targets and silences protein tyrosine phosphatase non-receptor type 13 (PTPN13) to maintain the activation of Src, a dephosphorylation substrate of PTPN13, thus reinforcing EGFR pathway in a regulatory circuit. miR-26a inhibition significantly improved NSCLC responses to gefitinib. These data revealed a novel mechanism of NSCLC resistance to TKI treatment.

TOPK promotes lung cancer resistance to EGFR tyrosine kinase inhibitors by phosphorylating and activating c-Jun.

Tyrosine kinase inhibitors (TKIs) targeting the epidermal growth factor receptor (EGFR) have shown promising clinical efficacy in non-squamous non-small cell lung cancer (NSCLC); however, resistance is frequently observed in malignant cells, operating through a mechanism that remains largely unknown. The present study shows that T-lymphokine-activated killer cell-originated protein kinase (TOPK) is upregulated in NSCLC and excessively activated in TKI-refractory cells. TOPK dictates the responsiveness of lung cancers to the EGFR-targeted TKI gefitinib through the transcription factor AP-1 component c-Jun. TOPK binds directly to and phosphorylates c-Jun, which consequently activates the transcription of AP-1 target genes, including CCND1 and CDC2. TOPK silencing sensitizes EGFR-TKI-resistant lung cancer cells to gefitinib and increases gefitinib efficacy in preclinical lung adenocarcinoma xenograft models. These findings represent a novel mechanism of lung cancer resistance to TKIs and suggest that TOPK may have value both as a predictive biomarker and as a therapeutic target: TOPK-targeted therapy may synergize with EGFR-targeted therapy in lung cancers.

Prognostic significance of G2/M arrest signaling pathway proteins in advanced non-small cell lung cancer patients.

The aim of the present study was to retrospectively assess the correlation between the expression levels of proteins involved in G2/M arrest signaling pathways in non-small cell lung cancer (NSCLC) tissue, as determined by immunohistochemical (IHC) methods, and the overall survival of patients with advanced stage NSCLC. IHC analysis of advanced NSCLC specimens was used to determine the expression levels of proteins involved in G2/M arrest signaling pathways, including ataxia telangiectasia mutated (ATM) kinase, ataxia telangiectasia and Rad3-related (ATR) kinase, checkpoint kinase (Chk) 1, Chk2, cell division cycle 25C (Cdc25C), total cyclin-dependent kinase 1 (Cdk1) and active Cdk1 signaling pathways, the latter of which refers to dephospho-Cdk1 (Tyr15) and phospho-Cdk1 (Thr161). Patients were enrolled continuously and followed up for ≥2 years. Univariate analysis demonstrated that the protein expression levels of dephospho-Cdk1 (P=0.015) and phospho-Cdk1 (P=0.012) exhibited prognostic significance, while the expression of the other proteins was not significantly associated with patient survival (ATM, P=0.843; ATR, P=0.245; Chk1, P=0.341; Chk2, P=0.559; Cdc25C, P=0.649; total Cdk1, P=0.093). Furthermore, the patients with tumors exhibiting low expression levels of active Cdk1 survived significantly longer than those with tumors exhibiting high active Cdk1 expression levels (P<0.05). In addition, Cox regression analysis demonstrated that the expression of active Cdk1 [odds ratio (OR), 0.624; 95% confidence ratio (CI), 0.400-0.973; P=0.038] and the pathological tumor-node-metastasis stage (OR, 0.515; 95% CI, 0.297-0.894; P=0.018) were significant independent prognostic factors for NSCLC. Therefore, the results of the present study indicated that active Cdk1 protein is an independent prognostic factor for advanced NSCLC and may validate Cdk1 as a therapeutic target for advanced NSCLC patients.

[Influence of ablactation rats' high calcium intake to the expression of uncoupling protein 2 mRNA after adulthood obesity induction].

OBJECTIVE: To study the influence of early high calcium intake to adulthood obesity through detection the expression of uncoupling protein 2 ( UCP2) mRNA in muscle by a reverse transcription polymerase chain reaction ( RT-PCR) technique. METHODS: 120 male Wistar rats were divided randomly into normal control group, high dose calcium group, medium dose calcium group and low doses calcium group, with basic diet and high calcium diet for 4 weeks. After intervention, blood fat was compared. Subsequently, all rats were fed basic diet for 3 weeks and blood fat was compared. Then, normal feed rats were randomly divided into normal controls and obesity induction group. Obesity induction group, high dose calcium group, medium dose calcium group and low doses calcium group were fed high-fat food. After 8 weeks, RT-PCR was used for analysis After obesity induction, weight the expression of UCP2 mRNA in muscle. RESULTS: growth of three high calcium groups were below obesity induction group, weight growth of low doses and high doses of high calcium groups were no difference with normal control group. Serum triglyceride levels of high dose high calcium group were significantly lower than obesity induction group and no difference with normal control group. Expression level of UCP2 mRNA of obesity induction group and high doses of high calcium group were obviously lower than normal controls, low dose and medium dose high calcium group, medium dose high calcium group' expression was significantly higher than normal control group. CONCLUSION: In rats' early life high calcium intake can continue to affect adulthood obesity induced by high-fat feed, increase expression level of UCP2 mRNA, improve the disorder of blood fat metabolism.