Functionalized graphene oxide NPs as a nanocarrier for drug delivery system in quercetin/ lurbinectedin as dual sensitive therapeutics for A549 lung cancer treatment.

Functionalized graphene oxide nanoparticles (NPs) have emerged as promising nanocarriers for drug delivery in lung cancer therapy. Quercetin and lurbinectedin encapsulated in graphene oxide (GO) NPs are tested for treating A549 lung cancer cells. Spectroscopic analyses show that graphene oxide functionalization creates a transparent, smooth surface for drug loading. Treatment with quercetin/lurbinectedin-loaded GO NPs induces notable cytotoxic effects in lung cancer cells, as evidenced by distinct morphological alterations and confirmed apoptotic cellular death observed through fluorescence microscopy. Additionally, our study highlights the impact of this approach on lung cancer metastasis, supported by qRT-PCR analysis of relative gene expression levels, including p53, Bax, Caspase-3, and Bcl 2, revealing robust molecular mechanisms underlying therapeutic efficacy against A549 and PC9 cell lines. Flow cytometric analyses further confirm the induction of cellular death in lung cancer cells following administration of the nanoformulation. Our findings show that quercetin/lurbinectedin-loaded GO NPs may be a promising lung cancer treatment, opening new avenues for targeted and effective therapies.

Evaluation of the Chinese versions of the Minnesota nicotine withdrawal scale and the questionnaire on smoking urges-brief.

OBJECTIVES: To evaluate the reliability and validity of the Chinese versions of Minnesota Nicotine Withdrawal Scale and Questionnaire on Smoking Urges-Brief (MNWS-C and QSU-Brief-C) in Chinese smokers. DESIGN AND METHODS: MNWS and QSU-Brief were translated into Chinese version through 10 steps. The reliability and validity of Chinese versions of the two scales were evaluated based on the data collected from 354 subjects. Cronbach's alpha coefficient was calculated to assess the reliability. Construct validity were evaluated with confirmatory factor analysis. Criteria validity was examined with correlation analysis between scale scores and patient-evaluated scores of craving and tobacco withdrawal symptoms. RESULTS: Cronbach's alpha coefficient of QSU-Brief-C was .96. Confirmatory factor analysis demonstrated that the fit indexes (adjusted goodness-of-fit index [AGFI], comparative fit index [CFI], normed fit index [NFI], and non-normed fit index [NNFI]) exceeded or approached 0.9. The correlation between QSU-Brief-C scores and patient-evaluated craving scores were statistically significant(r = .75, p < .0001). Cronbach's alpha coefficient of MNWS-C was .90. Confirmatory factor analysis demonstrated that the fit indexes (AGFI, CFI, NFI, and NNFI) exceeded 0.9. The correlation between MNWS-C scores and patient-evaluated discomfort scores were statistically significant(r = .68, p < .0001). CONCLUSION: Both QSU-Brief-C and MNWS-C have satisfactory validity and reliability and retain the two dimensions identified in their corresponding original English versions, which suggest that QSU-Brief-C and MNWS-C can be used in further research and clinical evaluation in Chinese smoking population with acceptable validity and reliability.

Long non-coding RNA SDPR-AS affects the development of non-small cell lung cancer by regulating SDPR through p38 MAPK/ERK signals.

We aimed to determine the roles and possible mechanism of long non-coding RNA SDPR-AS in the processes of non-small cell lung cancer (NSCLC). The expressions of SDPR-AS and SDPR in different subtypes of lung cancer (AC, SCC, LCC and SCLC) tissues and cells were determined. Three NSCLC cells were infused with pcDNA-SDPR-AS, pcDNA3.1, sh-SDPR-AS, sh-NC, si-SDPR and si-NC. The effects of SDPR-AS dysregulation on cell behaviors (including cell viability, colony forming ability, migration, invasion and apoptosis) were assessed. Moreover, the combined effects of SDPR-AS overexpression and SDPR-AS knockdown on H522 cell behaviors and the levels of p-p38 and p-ERK were investigated. SDPR-AS was lowly expressed in NSCLC tissues and cells, but had no changes in SCLC tissues and cells. Down-regulation of SDPR-AS enhanced the proliferation, migration and invasion and inhibited apoptosis of NSCLC cells (H522, H661 and H520). Overexpression of SDPR-AS exhibited opposite effects. Moreover, SDPR was positively regulated by SDPR-AS and effects of SDPR-AS on the cell biological processes of NSCLC cells were through regulation of SDPR. Besides, the levels of p-p38 and p-ERK were significantly decreased after SDPR-AS overexpression, which were dramatically changeover by SDPR knockdownsimultaneously. Our findings indicate that SDPR-AS was lowly expressed in NSCLC cells and down-regulation of SDPR-AS may promote the malignant behaviors of NSCLC cells possible through regulating SDPR expression and involving in p38 MAPK/ERK signaling pathway. SDPR-AS may serve as a prospective target for NSCLC diagnosis and therapy.

Lentivirus-mediated overexpression of suppressor of cytokine signaling-3 reduces neutrophilic airway inflammation by suppressing T-helper 17 responses in mice with chronic Pseudomonas aeruginosa lung infections.

The aim of the present study was to explore the effect of overexpressed suppressor of cytokine signaling­3 (SOCS3) on T-helper (Th)17 cell responses and neutrophilic airway inflammation in mice with chronic Pseudomonas aeruginosa (PA) infections. SOCS3 expression was enhanced via the administration of tail vein injections of therapeutic lentivirus in mice with chronic PA lung infections. SOCS3 expression in the blood and lung tissue was assessed using reverse transcription­quantitative polymerase chain reaction (RT­qPCR) and western blot analysis. Total and differential cell numbers and myeloperoxidase levels in the bronchoalveolar lavage (BAL) fluid were assessed, as well as the number of bacterial colonies in the lungs. Histological analysis of lung tissue was performed using hematoxylin and eosin staining and phosphorylated­signal transducer and activator of transcription­3 (p­STAT3) expression was measured by western blot analysis and immunohistochemistry. The expression of STAT3 mRNA and retinoid­related orphan receptor (ROR)γt were measured by RT­qPCR. The percentage of interleukin (IL)­17+ cells among cluster of differentiation (CD)4+ cells was calculated using flow cytometry and levels of IL­17A and IL­6 were assessed by ELISA. The expression of SOCS3 was significantly increased in CD4+ T cells following lentivirus injection and the inflammation of neutrophilic airways was notably ameliorated. Enhanced SOCS3 expression was associated with a significant decrease in the expression of p­STAT3 and RORγt in CD4+ T cells. Additionally, the percentage of IL­17+ cells among CD4+ T cells and the IL­17 contents in the BAL fluid were significantly decreased. Lentivirus­mediated overexpression of SOCS3 was revealed to ameliorate neutrophilic airway inflammation by inhibiting pulmonary Th17 responses in mice with chronic PA lung infections.

Plasma follistatin-like protein 1 is correlated with disease severity in patients with acute pulmonary embolism and predicts short-term mortality.

We investigated the plasma levels of follistatin-like protein 1 (FSTL1) in patients with acute pulmonary embolism (PE) and whether it could predict short-term mortality. A prospective observational cohort study was conducted in patients with acute PE (n = 220). FSTL1 was measured in plasma samples using enzyme-linked immunosorbent assay. Plasma FSTL1 levels were significantly increased in patients with acute PE, and positively correlated with disease severity (rs = 0.7171). Sensitivity and specificity rates for high-risk PE at a specific FSTL1 cutoff point (23 ng/ml) were 79.3% and 92.9%. Multivariant Cox regression analysis showed that FSTL1 was independently associated with 30-day mortality rate. Addition of FSTL1 to the PE severity index (PESI) scoring system significantly improved the predictive value for 30-day mortality. These results indicate that the plasma level of FSTL1 is correlated with disease severity in patients with acute PE and predicts short-term mortality.

Upregulation of SOCS3 in lung CD4+ T cells in a mouse model of chronic PA lung infection and suppression of Th17­mediated neutrophil recruitment in exogenous SOCS3 transfer in vitro.

Neutrophilic airway inflammation in chronic lung infections caused by Pseudomonas aeruginosa (PA) is associated with T helper (Th)17 responses. Suppressor of cytokine signaling 3 (SOCS3) is the major negative modulator of Th17 function through the suppression of signal transducer and activator of transcription (STAT)3 activation. The aim of the present study was to investigate the expression of SOCS3 in lung CD4+ T cells in a mouse model of chronic PA lung infection and the effect of exogenous SOCS3 on Th17­mediated neutrophil recruitment in vitro. A mouse model of chronic PA lung infection was established and the activation of STAT3 and Th17 response in lung tissues and lung CD4+ T cells was assessed. The protein and mRNA expression of SOCS3 in lung CD4+ T cells was analyzed by western blotting and reverse transcription­quantitative polymerase chain reaction. The authors constructed a recombinant lentivirus carrying the SOCS3 gene and transferred it into lung CD4+ T cells isolated from a mouse model. These transfected cells were stimulated with interleukin (IL)­23 in vitro and the protein level of p­STAT3 and retinoid­related orphan receptor (ROR)γt was determined by western blotting. The expression of IL­17A+ cells was analyzed by flow cytometry and the level of IL­17A in cell culture supernatant was measured by ELISA. The mouse lung epithelial cell line, MLE­12, was cocultured with lung CD4+ T cells that overexpressed the SOCS3 gene and the culture supernatant was harvested and used for a chemotaxis assay. Compared with control mice, mice with chronic PA lung infection had significantly higher level of p­STAT3 and Th17 response in both lung tissues and lung CD4+ T cells. The protein and mRNA level of SOCS3 in lung CD4+ T cells increased as the chronic PA lung infection developed. Exogenous SOCS3 gene transfer in PA­infected lung CD4+ T cells decreased p­STAT3 and RORγt expression and suppressed the level of IL­17A+ cells in vitro. MLE­12 cells cocultured with SOCS3­overexpressing lung CD4+ T cells expressed a significantly lower level of neutrophil chemoattractants chemokine (C­X­C motif) ligand (CXCL) 1 and CXCL5, and recruited significantly smaller numbers of migrating neutrophils than those cocultured with control cells. SOCS3 was upregulated in lung CD4+ T cells following the activation of STAT3/Th17 axis in a mouse model of chronic PA lung infection. Exogenous SOCS3 transfer in PA­infected lung CD4+ T cells suppresses Th17­mediated neutrophil recruitment in vitro.

Follistatin-like protein 1 is elevated in systemic autoimmune diseases and correlated with disease activity in patients with rheumatoid arthritis.

INTRODUCTION: Follistatin-like protein 1 (FSTL1) is a proinflammation mediator implicated in arthritis in rodent animal models. The present study is aimed at assessing FSTL1 levels in systemic autoimmune diseases and correlating them with disease activity in patients with rheumatoid arthritis (RA). METHODS: Serum FSTL1 levels from 487 patients with systemic autoimmune diseases and 69 healthy individuals were measured by enzyme-linked immunosorbent assay (ELISA). FSTL1 expression in synovial fluid (SF) and synovial tissues (STs) was determined by ELISA, immunohistochemistry, real-time polymerase chain reaction (RT-PCR) and western blot analysis in RA patients and trauma controls. FSTL1 levels in fibroblast-like synoviocytes (FLSs) from RA patients were determined by real-time PCR and western blot analysis. RESULTS: Serum FSTL1 levels were significantly elevated in patients with RA, ulcerative colitis, systemic lupus erythematosus, Sjögren's syndrome (SS), systemic sclerosis and polymyositis/dermatomyositis. Serum FSTL1 levels in the RA and secondary SS patients were substantially higher than those in other patients. Serum FSTL1 levels were increased in early RA, rheumatoid factor (RF)- and anti-cyclic citrullinated peptide antibody (ACPA)-negative patients compared to healthy controls. Moreover, serum FSTL1 concentrations were significantly higher in long-standing RA patients than in early RA patients and in the RF- and ACPA-positive RA patients than in RF- and ACPA-negative RA patients. Elevated FSTL1 levels in the STs and SF of RA patients were also observed. FSTL1 levels in serum were markedly higher than those in SF in RA patients. The strongest FSTL1 staining was detected in the cytoplasm of synovial and capillary endothelial cells from RA synovium. Furthermore, FSTL1 was induced in FLSs by inflammatory mediators. Importantly, serum FSTL1 levels were correlated with several important biologic and clinical markers of disease activity, including erythrocyte sedimentation rate, C-reactive protein, RF, ACPA, swollen joint count, patient global visual analogue scale score and Disease Activity Score 28 in the adult RA patient population. Notably, serum FSTL1 levels were significantly diminished following successful treatment and clinical improvement. CONCLUSIONS: Elevated FSTL1 levels reflect not only joint diseases but also inflammation and tissue degradation in systemic autoimmune diseases. Serum FSTL1 levels may thus serve as a serological inflammatory marker of disease activity in RA patients.