Adaptive promising zone design for cancer immunotherapy with heterogeneous delayed treatment effect.

Indirect mechanisms of cancer immunotherapies result in delayed treatment effects that vary among patients. Consequently, the use of the log-rank test in trial design and analysis can lead to significant power loss and pose additional challenges for interim decisions in adaptive designs. In this paper, we describe patients' survival using a piecewise proportional hazard model with random lag time and propose an adaptive promising zone design for cancer immunotherapy with heterogeneous delayed effects. We provide solutions for calculating conditional power and adjusting the critical value for the log-rank test with interim data. We divide the sample space into three zones - unfavourable, promising, and favourable -based on re-estimations of the survival parameters, the log-rank test statistic at the interim analysis, and the initial and maximum sample sizes. If the interim results fall into the promising zone, the sample size is increased; otherwise, it remains unchanged. We show through simulations that our proposed approach has greater overall power than the fixed sample design and similar power to the matched group sequential trial. Furthermore, we confirm that critical value adjustment effectively controls the type I error rate inflation. Finally, we provide recommendations on the implementation of our proposed method in cancer immunotherapy trials.

Electrochemiluminescence imaging of a membrane carcinoembryonic antigen at single tissue sections.

Histopathological molecular testing of tissue sections is an essential step in tumor diagnosis; however, the commonly used immunohistochemical methods have problems such as low specificity and the subjective bias of the observer. Here, we report an electrochemiluminescence (ECL) imaging method to detect a membrane carcinoembryonic antigen (CEA) at the single tissue sections of cancer patients. By permeabilizing the tissue attached to a glassy carbon electrode, Ru(bpy)32+ tagged at the membrane CEA of the tissue could electrochemically react with TPrA in solution to emit ECL that has near-zero background and an extremely high signal-to-background ratio. Using the established ECL method, the expression differences and distribution characteristics of the CEA protein in the carcinoma and paracancerous tissues of pancreatic ductal carcinoma (PDAC) and lung adenocarcinoma (LUAD) patients are investigated. The images reveal that CEA proteins are mostly distributed in the acini and surrounding areas both in PDAC and LUAD tissues. Therefore, the presented approach could be able to provide a new molecular recognition method for the diagnosis of adenocarcinoma and other tumors.

Risk of waning humoral responses after inactivated or subunit recombinant SARS-CoV-2 vaccination in patients with chronic diseases: Findings from a prospective observational study in China.

Heterogeneity of antibody responses has been reported in SARS-CoV-2 vaccination recipients with underlying diseases. We investigated the impact of the presence of comorbidities on the humoral response to SARS-CoV-2 vaccination in patients with chronic disease (PWCD) and assessed the effect of the number of comorbidities on the humoral response to vaccination. In this study, neutralizing antibodies (NAbs) and IgG antibodies against the receptor-binding domain (RBD-IgG) were monitored following a full-course vaccination. In total, 1400 PWCD (82.7%, inactivated vaccines; 17.3%, subunit recombinant vaccine) and 245 healthy controls (65.7% inactivated vaccines, 34.3% subunit recombinant vaccine) vaccinated with inactivated or subunit recombinant SARS-CoV-2 vaccines, were included. The seroconversion and antibody levels of the NAbs and RBD-IgG were different in the PWCD group compared with those in the control group. Chronic hepatitis B (odds ratio [OR]: 0.65; 95% confidence interval [CI]: 0.46-0.93), cancer (OR: 0.65; 95% CI: 0.42-0.99), and diabetes (OR: 0.50; 95% CI: 0.28-0.89) were associated with lower seroconversion of NAbs. Chronic kidney disease (OR: 0.29; 95% CI: 0.11-0.76), cancer (OR: 0.38; 95% CI: 0.23-0.62), and diabetes (OR: 0.37; 95% CI: 0.20-0.69) were associated with lower seroconversion of RBD-IgG. Only the presence of autoimmune disease showed significantly lower NAbs and RBD-IgG titers. Patients with most types of chronic diseases showed similar responses to the controls, but humoral responses were still significantly associated with the presence of ≥2 coexisting diseases. Our study suggested that humoral responses following SARS-CoV-2 vaccination are impaired in patients with certain chronic diseases.

High-Coverage Serum Metabolomics Reveals Metabolic Pathway Dysregulation in Diabetic Retinopathy: A Propensity Score-Matched Study.

Background: Diabetic retinopathy (DR) is a major diabetes-related disease linked to metabolism. However, the cognition of metabolic pathway alterations in DR remains scarce. We aimed to corroborate alterations of metabolic pathways identified in prior studies and investigate novel metabolic dysregulations that may lead to new prevention and treatment strategies for DR. Methods: In this case-control study, we tested 613 serum metabolites in 69 pairs of type 2 diabetic patients (T2DM) with DR and propensity score-matched T2DM without DR via ultra-performance liquid chromatography-tandem mass spectrometry system. Metabolic pathway dysregulation in DR was thoroughly investigated by metabolic pathway analysis, chemical similarity enrichment analysis (ChemRICH), and integrated pathway analysis. The associations of ChemRICH-screened key metabolites with DR were further estimated with restricted cubic spline analyses. Results: A total of 89 differentially expressed metabolites were identified by paired univariate analysis and partial least squares discriminant analysis. We corroborated biosynthesis of unsaturated fatty acids, glycine, serine and threonine metabolism, glutamate and cysteine-related pathways, and nucleotide-related pathways were significantly perturbed in DR, which were identified in prior studies. We also found some novel metabolic alterations associated with DR, including the disturbance of thiamine metabolism and tryptophan metabolism, decreased trehalose, and increased choline and indole derivatives in DR. Conclusions: The results suggest that the metabolism disorder in DR can be better understood through integrating multiple biological knowledge databases. The progression of DR is associated with the disturbance of thiamine metabolism and tryptophan metabolism, decreased trehalose, and increased choline and indole derivatives.

Comparison of CRP, Procalcitonin, Neutrophil Counts, Eosinophil Counts, sTREM-1, and OPN between Pneumonic and Nonpneumonic Exacerbations in COPD Patients.

Introduction: The patients with community-acquired pneumonia (CAP) and acute exacerbations of COPD (AECOPD) could have a higher risk of acute and severe respiratory illness than those without CAP in AECOPD. Consequently, early identification of pneumonia in AECOPD is quite important. Methods. 52 subjects with AECOPD + CAP and 93 subjects with AECOPD from two clinical centers were enrolled in this prospective observational study. The values of osteopontin (OPN), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), C-reactive protein (CRP), procalcitonin (PCT), eosinophil (EOS) counts, and neutrophil (Neu) counts in blood on the first day of admission and clinical symptoms were compared in AECOPD and AECOPD + CAP. In addition, subgroup analyses of biomarker difference were conducted based on the current use of inhaled glucocorticoids (ICS) or systemic corticosteroids (SCS). Results: Patients with AECOPD + CAP had increased sputum volume, sputum purulence, diabetes mellitus, and longer hospital stays than AECOPD patients (p < 0.05). A clinical logistic regression model showed among the common clinical symptoms, purulent sputum can independently predict pneumonia in AECOPD patients after adjusting for a history of diabetes. At day 1, AECOPD + CAP patients had higher values of Neu, CRP, PCT, and OPN, while serum sTREM-1 levels and EOS counts were similar in the two groups. CRP fared best at predicting AECOPD with CAP (p < 0.05 for the test of difference), while OPN had similar accuracy with Neu, PCT, and purulent sputum (p > 0.05 for the test of difference). Multivariate analysis, including clinical symptoms and biomarkers, suggested that CRP ≥15.8 mg/dL at day 1 was a only promising predictor of pneumonia in AECOPD. CRP and OPN were not affected by ICS or SCS. Conclusions: CRP ≥15.8 mg/dL is an ideal promising predictor of pneumonia in AECOPD, and its plasma level is not affected by ICS or SCS. The diagnostic performance of CRP is not significantly improved when combined with clinical symptoms or other markers (OPN, PCT, and Neu).

Chordin-like 1 is a novel prognostic biomarker and correlative with immune cell infiltration in lung adenocarcinoma.

Chordin-like 1 (CHRDL1), an inhibitor of bone morphogenetic proteins(BMPs), has been recently reported to participate in the progression of numerous tumors, however, its role in lung adenocarcinoma (LUAD) remains unclear. Our study aimed to demonstrate relationship between CHRDL1 and LUAD based on data from The Cancer Genome Atlas (TCGA). Among them, CHRDL1 expression revealed promising power for distinguishing LUAD tissues form normal sample. Low CHRDL1 was correlated with poor clinicopathologic features, including high T stage (OR=0.45, P<0.001), high N stage (OR=0.57, P<0.003), bad treatment effect (OR=0.64, P=0.047), positive tumor status (OR=0.63, P=0.018), and TP53 mutation (OR=0.49, P<0.001). The survival curve illustrated that low CHRDL1 was significantly correlative with a poor overall survival (HR=0.60, P<0.001). At multivariate Cox regression analysis, CHRDL1 remained independently correlative with overall survival. GSEA identified that the CHRDL1 expression was related to cell cycle and immunoregulation. Immune infiltration analysis suggested that CHRDL1 was significantly correlative with 7 kinds of immune cells. Immunohistochemical validation showed that CHRDL1 was abnormally elevated and negatively correlated with Th2 cells in LUAD tissues. In conclusion, CHRDL1 might become a novel prognostic biomarker and therapy target in LUAD. Moreover, CHRDL1 may improve the effectiveness of immunotherapy by regulating immune infiltration.

Ultrasound-mediated nanobubble destruction (UMND) facilitates the delivery of VEGFR2-targeted CD-TK-loaded cationic nanobubbles in the treatment of bladder cancer.

BACKGROUND: The CD-TK double suicide gene has become an effective therapy for bladder cancer. A novel molecular-targeted ultrasound (US) method has been developed to precisely guide nanobubbles loaded with this gene to regions within bladder tumor cells and is widely used due to its efficiency in delivering drugs to the target tumor. METHODS: Uniform nanoscaled nanobubbles loaded with CD-TK double suicide gene were developed using a thin-film hydration sonication, carbodiimide chemistry approaches, and electrostatic adsorption methods. RESULTS: In the present study, we synthesized CD-TK double suicide gene-loaded cationic nanobubbles conjugated with anti-VEGFR2 that can bind with VEGFR2-positive cells. Fluorescence and flow cytometry evidence show that CD-TK double suicide gene-loaded nanobubbles were successfully developed. CD-TK-CNBs delivered via US-mediated nanobubble destruction (UMND) enhanced transfection efficiency, overexpression of CD-TK double suicide gene, and tumor cell apoptosis, and inhibited tumor cell growth in vitro. CONCLUSIONS: These CD-TK-CNBs may become a novel treatment for bladder cancer.

Prostate-Specific Antigen Test Utilization in a Major Canadian City.

OBJECTIVES: To review the utilization of prostate-specific antigen (PSA) testing in Winnipeg, a major Canadian city, and to compare PSA testing rates between Winnipeg and Calgary, another major Canadian city of comparable size. METHODS: PSA testing results were reviewed by year and age group. We focused our studies in years 2011 and 2016, for which census demographic data are available. RESULTS: In Winnipeg, the PSA testing rates (patients with one or two PSA tests divided by the male population) showed a declining trend over years from 2008 to 2017. For almost all age groups, PSA testing rates in 2016 decreased in comparison to those in 2011. For age older than 40 years, the relative percentage decreases were 14% to 20%.In 2011, Winnipeg PSA testing rates were consistently higher than those in Calgary for all age groups. For age older than 40 years, the relative percentage differences were 36% to 50%.In addition, 41% and 40% of patients in Winnipeg who underwent PSA testing were younger than 50 years or older than 69 years in 2011 and 2016, respectively. CONCLUSIONS: PSA testing utilization may be falling short of optimal rates. There is a need to reinforce the optimal use of clinical recommendations.

TMEM16A as a Potential Biomarker in the Diagnosis and Prognosis of Lung Cancer.

BACKGROUND: Transmembrane protein 16A (TMEM16A), also known as ANO1 (anoctamin-1), was reported to be vital in the growth and invasion of several malignancies. However, role of TMEM16A in lung cancer remained unclear. The aim of this study was to evaluate the expression of TMEM16A and its significance in lung cancer. METHODS: qRT-PCR and Western blots were performed to evaluate the TMEM16A mRNA and protein expression. Proliferation and invasion of H1299 cancer cells were evaluated by CCK-8 and transwell assays. Tumor volumes in nude mice implanted with H1299 cells were assessed once every week for 5 weeks by measuring 2 perpendicular dimensions. Immunofluorescent staining revealed expression of TMEM16A in nude mice cancer tissues. RESULTS: Our findings provided compelling evidence that TMEM16A production in H1299 cells is 2.1 times higher than observations in HBE16 cells. We showed that overexpression of TMEM16A contributed to the proliferation of H1299 cells. Moreover, T16Ainh-A01, a specific TMEM16A inhibitor or shRNA targeting TMEM16A somewhat inhibited lung tumor cell growth and invasion as evident from in vitro studies and from in vivo xenograft-tumor growth. Inhibition of TMEM16A strongly suppressed EGFR phosphorylation and growth of lung cancer cells. Furthermore, a reduction of p-RAS and p-ERK1/2 was also observed. CONCLUSION: TMEM16A promoted growth and invasion in lung cancer cells via an EGFR/ MAPK-dependent signaling pathway. So we infer TMEM16A membrane protein may have potential to serve as a biomarker in lung cancer.

The effect of timing of oseltamivir chemoprophylaxis in controlling influenza A H3N2 outbreaks in long-term care facilities in Manitoba, Canada, 2014-2015: a retrospective cohort study.

OBJECTIVE: This study examined the effect of the timing of administration of oseltamivir chemoprophylaxis for the control of influenza A H3N2 outbreaks among residents in long-term care facilities (LTCFs) in Manitoba, Canada, during the 2014-2015 influenza season. METHODS: A retrospective cohort study was conducted of all LTCF influenza A H3N2 outbreaks (n=94) using a hierarchical logistic regression analysis. The main independent variable was how many days passed between the start of the outbreak and commencement of oseltamivir chemoprophylaxis. The dependent variable was whether each person in the institution developed influenza-like illness (yes or no). RESULTS: Delay of oseltamivir chemoprophylaxis was associated with increased odds of infection in both univariate (t=5·41; df=51; P<·0001) and multivariable analyses (t=6·04; df=49; P<·0001) with an adjusted odds ratio of 1.3 (95% confidence interval [CI], 1·2-1·5) per day for influenza A H3N2. CONCLUSIONS: The sooner chemoprophylaxis is initiated, the lower the odds of secondary infection with influenza in LTCFs during outbreaks caused by influenza A H3N2 in Manitoba. For every day that passed from the start of the outbreak to the initiation of oseltamivir, the odds of a resident at risk of infection in the facility developing symptomatic infection increased by 33%.

Plasma YKL-40 and NGAL are useful in distinguishing ACO from asthma and COPD.

BACKGROUND: Asthma-chronic obstructive pulmonary disorder (COPD) overlap (ACO) is characterized by the coexistence of features of both asthma and COPD and is associated with rapid progress and a poor prognosis. Thus, the early recognition of ACO is crucial. OBJECTIVES: We sought to explore the plasma levels of biomarkers associated with asthma (periostin, TSLP and YKL-40), COPD (NGAL) and their possible correlation with lung function, the bronchodilator response and radiographic imaging in patients with asthma, COPD and with features of ACO. METHODS: We enrolled 423 subjects from 6 clinical centers. All participants underwent blood collection, lung function measurements, bronchodilator response tests and high-resolution CT. Correlations of the plasma biomarkers with lung function, the bronchodilator response and percentemphysema were calculated by Spearman's rank correlation and multivariate stepwise regressionanalysis. RESULTS: 1) Patients with features of ACO had lower plasma YKL-40 than COPD patients and a moderate elevated plasma level of NGAL compared with asthma patients. 2) Patients with features of ACO had an intermediate degree of airflow obstruction, the bronchodilator response and emphysema between patients with COPD and asthma. 3) Plasma YKL-40 was negatively correlated with lung function and with the bronchodilator response, and plasma NGAL was positively correlated with the extent of emphysema. CONCLUSIONS: Plasma YKL-40 is a promising candidate for distinguishing between patients with features of ACO and COPD patients, while plasma NGAL may be a valuable biomarker for differentiating between patients with features of ACO and asthma patients. CLINICAL TRIAL REGISTRATION: ChiCTR-OOC-16009221.

Interleukin-13 stimulates MUC5AC expression via a STAT6-TMEM16A-ERK1/2 pathway in human airway epithelial cells.

Transmembrane protein 16A (TMEM16A), a channel underlying the calcium-activated chloride channel (CaCC) currents, has been shown to be a key regulator of mucus overproduction in airway epithelial cells. However, the precise molecular mechanism involved in the TMEM16A-mediated mucus secretion remains unclear. In the present study, we inquired into a novel signaling mechanism for TMEM16A driving mucin 5AC (MUC5AC) production in human airway epithelial cells. Following treatment for 24-48h with type 13 interleukin (IL-13), an upregulation of TMEM16A expression in both mRNA and protein levels was observed in human bronchial epithelial cell line (HBE16), while signal transducer and activator of transcription 6 (STAT6) inhibition could decrease this elevated expression, suggesting that the regulation of TMEM16A expression by IL-13 was via a STAT6-based transcriptional mechanism. Further investigation of the HBE16 cells revealed that TMEM16A knockdown or specific chloride channel inhibitor T16Ainh-A01 could suppress the CaCC currents and consequently reduce the extracellular regulated kinase (ERK1/2) phosphorylation, accompanying a dramatical decrease in MUC5AC expression. Moreover, pretreated with PD98059, an inhibitor of ERK1/2, the HB16 cells showed a remarkable diminution in TMEM16A-mediated MUC5AC expression. Altogether, STAT6-TMEM16A-ERK1/2 signal pathway and TMEM16A channel activity are required for the IL-13-induced TMEM16A mediated mucus production.

TMEM16A mediates the hypersecretion of mucus induced by Interleukin-13.

Previous studies showed that the Ca(2+)-activated Cl(-) channel (CaCC) was involved in the pathogenesis of mucus hypersecretion induced by Interleukin-13 (IL-13). However, the mechanisms underlying the process were unknown. Recently, transmembrane protein 16A (TMEM16A) was identified as the channel underlying the CaCC current. The aim of the current study was to investigate whether the TMEM16A channel is part of the mechanism underlying IL-13-induced mucus hypersecretion. We observed that both TMEM16A mRNA and protein expression were significantly up-regulated after treatment with IL-13 in human bronchial epithelial 16 (HBE 16) cells, which correlated with an increase in mucus production. Additionally, mucus hypersecretion in rat airways was induced by intratracheal instillation of IL-13 and similar increases were observed in the expression of TMEM16A mRNA and protein in the bronchial epithelium. Niflumic acid (NA), a selective antagonist of CaCC, markedly blocked IL-13-induced mucin (MUC) 5AC mRNA and protein production in vivo and in vitro. Further investigation with HBE16 cells revealed that TMEM16A overexpression clearly promoted mucus production, IκBα phosphorylation, and p65 accumulation in the nucleus. The loss of TMEM16A resulted in inhibition of mucus production, and the TMEM16A-mediated production of MUC5AC was significantly blocked by a nuclear factor-kappa B (NF-κB) inhibitor. Therefore, the TMEM16A channel acts upstream of NF-κB in the regulation of mucus production. This is the first demonstration that the TMEM16A-NF-κB pathway is positively involved in IL-13-induced mucus production, which provides novel insight into the molecular mechanism of mucin overproduction.

Effects of lifestyle intervention on dietary intake, physical activity level, and gestational weight gain in pregnant women with different pre-pregnancy Body Mass Index in a randomized control trial.

BACKGROUND: The objectives of this study were to assess the efficacy of lifestyle intervention on gestational weight gain in pregnant women with normal and above normal body mass index (BMI) in a randomized controlled trial. METHODS: A total of 116 pregnant women (<20 weeks of pregnancy) without diabetes were enrolled and 113 pregnant women completed the program. Participants were randomized into intervention and control groups. Women in the intervention group received weekly trainer-led group exercise sessions, instructed home exercise for 3-5-times/week during 20-36 weeks of gestation, and dietary counseling twice during pregnancy. Participants in the control group did not receive the intervention. All participants completed a physical activity questionnaire and a 3-day food record at enrolment and 2 months after enrolment. RESULTS: The participants in the intervention group with normal pre-pregnancy BMI (≤24.9 kg/M2, n = 30) had lower gestational weight gain (GWG), offspring birth weight and excessive gestational weight gain (EGWG) on pregnancy weight gain compared to the control group (n = 27, p < 0.05). Those weight related-changes were not detected between the intervention (n = 27) and control group (n = 29) in the above normal pre-pregnancy BMI participants. Intervention reduced total calorie, total fat, saturated fat and cholesterol intake were detected in women with normal or above normal pre-pregnancy BMI compared to the control group (p < 0.05 or 0.01). Increased physical activity and reduced carbohydrate intake were detected in women with normal (p < 0.05), but not above normal, pre-pregnancy BMI at 2 months after the onset of the intervention compared to the control group. CONCLUSION: The results of the present study demonstrated that the lifestyle intervention program decreased EGWG, GWG, offspring birth weight in pregnant women with normal, but not above normal, pre-pregnancy BMI, which was associated with increased physical activity and decreased carbohydrate intake. TRIAL REGISTRATION: NCT00486629.

Risk Assessment and Clinical Risk Management for Young Antisocial Children: The Forgotten Group / Evaluación del riesgo y manejo del riesgo clínico en jóvenes antisociales: el grupo olvidado

Centre for Children Committing Offences (CCCO), at Child Development Institute (CDI) in Toronto, Canada, developed Early Assessment Risk Lists (EARL-20B for boys; EARL-21G for girls), for young children at-risk for future criminality. In this first EARL prospective longitudinal study, 573 boys and 294 girls who participated in SNAP®, a gender-specific evidence-based model for at-risk children (6-11 years), 8.2% of boys and 3.1% of girls had registered criminal offences at follow up (mean age 14.9 and 14.6 respectively). EARL Total, Family, Child, and Responsivity domain scores, including two gender-specific risk items and Overall Clinical Judgment predicted early onset of criminal activity. Findings suggest that gender-sensitive clinical risk assessment and management tools are important for effectively identifying and potentially reducing criminal outcomes.

En el Centro para Niños Infractores (CCCO), del Instituto de Desarrollo Infantil (CDI) en Toronto (Canadá), se desarrollaron las Listas de Evaluación de Riesgos Tempranos (EARL-20B para niños; EARL-21G para niñas), para niños en riesgo de desarrollar criminalidad. En este primer estudio longitudinal de las EARL, 573 niños y 294 niñas que participaron en SNAP un modelo basado en evidencia de género específico para riesgo en niños (6-11 años), 8.2 % de niños y 3.1 % de las niñas registraron delitos criminales durante el seguimiento (M = 14.9 y 14.6, respectivamente). Los puntajes de EARL Total, Familia, Niños y Responsividad, incluyendo dos ítems de riesgo específicos de género, y el Juicio Clínico General predicen el inicio temprano de actividad criminal. Los resultados sugieren que la evaluación del riesgo clínico sensible al género y el manejo de herramientas son importantes para la identificación efectiva y potencialmente reducen los resultados criminales.

The inhibition of aldose reductase on mucus production induced by interleukin-13 in the human bronchial epithelial cells.

This study investigated whether aldose reductase (AR) inhibition affects interleukin (IL)-13-induced mucus production in the human bronchial epithelial cell line-16 (HBE16) cells. The HBE16 cells were cultured with AR inhibitors (zopolrestat) or were transfected with an AR small interfering (si)RNA. Subsequently, the cells were stimulated with 10 ng/ml IL-13 for 2h. The levels of mucin (MUC)5AC mRNA and protein were measured by using RT-PCR or ELISA. Intracellular reactive oxygen species (ROS) were measured fluorimetrically with the CM-H2DCFDA probe. Western blotting was performed to determine the levels of AR, phosphorylated signal transducer and activator of transcription 6 (p-STAT6) and phosphorylated Janus kinase 2 (p-JAK2). The results show that treatment with zopolrestat or transfection with AR siRNA significantly suppressed IL-13-stimulated MUC5AC mRNA and protein in the HBE16 cells (P<0.05). AR inhibition could suppress IL-13-induced ROS generation, the phosphorylation of JAK2/STAT6 pathway and the activation of nuclear factor (NF)-kappa B, thereby decreasing mucus production in vitro (all P<0.05). Therefore, the inhibition of AR could be a therapeutic target for mucus hypersecretion in chronic inflammation lung disease.

Effects of scutellarin on MUC5AC mucin production induced by human neutrophil elastase or interleukin 13 on airway epithelial cells.

Scutellarin is a flavonoid extracted from a traditional Chinese herb, Erigeron breviscapus. The present study investigated the effect of scutellarin on MUC5AC mucin production and the possible mechanism. Human bronchial epithelial 16 (HBE16) cells were pretreated with scutellarin for 60 min, and then exposed to human neutrophil elastase (HNE) or interleukin (IL)-13 for 12 hr. RT-PCR and ELISA were performed to measure the amount of MUC5AC mucin production. The results showed that scutellarin inhibited MUC5AC expression both in mRNA and protein level induced by HNE in a concentration-dependent manner. However, scutellarin failed to inhibit MUC5AC mucin production induced by IL-13. To investigate the intracellular mechanisms associated with the effect of scutellarin on MUC5AC mucin production, western blotting was carried out to examine the phosphorylation of protein kinase C (PKC), signal transducer and activator of transcription 6 (STAT6) and extracellular signal-regulated kinase 1/2 (ERK1/2). The phosphorylation of PKC and ERK1/2 was attenuated after treatment with scutellarin, whereas STAT6 was not significantly affected. Therefore, it is suggested that scutellarin down-regulates MUC5AC mucin production on HBE16 cells via ERK-dependent and PKC-dependent pathways.

Feasibility and safety of adhesiolysis using transgastric NOTES approach: a pilot survival study in a porcine model.

OBJECTIVES: Although adhesions can be removed by adhesiolysis using laparotomy or laparoscopy, they typically recur sometimes with equal severity. It is suggested that minimizing the invasiveness of the operative technique by using natural orifice translumenal endoscopic surgery (NOTES) may reduce adhesion re-formation. The aim of the study was to evaluate the feasibility and safety of adhesiolysis by using a novel transgastric NOTES approach and collect pilot data on adhesion recurrence after transgastric NOTES adhesiolysis. METHODS: One nonsurvival and 5 survival female pigs were used in this experimental survival study. Interventions included (a) induction of adhesions by laparotomy, (b) 2 weeks survival, (c) transgastric NOTES adhesiolysis with endoscopic evaluation and scoring of adhesions before and immediately after adhesiolysis, (d) 2 weeks survival, and (e) necropsy with endoscopic and necroscopic evaluation and scoring of recurrent adhesions. Main outcome measures were (a) survival and complication rates and (b) assessment of adhesion formation and re-formation using the Hopkins Adhesion Formation Score. RESULTS: No mortality and no complications were observed. A total of 11 adhesions formed before the adhesiolysis in 5 survival study animals. All were successfully divided. The frequency of adhesions and median adhesion formation score decreased significantly immediately after adhesiolysis compared with that prior to the procedure (frequency, 11 vs 0, P = .011; the median score = 2.0 [range 1-3] vs 0.0 [range 0-0], P = .004). The treatment gains maintained at 2 weeks after the adhesiolysis. LIMITATIONS: The limitations of this study were the low number of study animals and short-term follow-up data. CONCLUSIONS: Adhesiolysis using NOTES transgastric approach is feasible, safe, and effective with minimal adhesion re-formation.

[Effects of secretary leukocyte protease inhibitor-transfected bone marrow mesenchymal stem cells on airway inflammation and mucus secretion in chronic obstructive pulmonary disease].

OBJECTIVE: To explore the effects of secretary leukocyte protease inhibitor (SLPI)-transfected bone marrow mesenchymal stem cells (BMSCs) transplantation on airway inflammation and mucus secretion in chronic obstructive pulmonary disease (COPD) rats. METHODS: Sixty rats were equally and randomly divided into negative control, COPD model, BMSCs and SLPI-transfected BMSCs groups. The COPD rat model was established in all rats with the exception of the negative control rats by smoking and intratracheal instillation of lipopolysaccharide (LPS). BMSCs with or without transfection of plasmid containing SLPI gene were delivered through caudal vein of rats at 30 days post-induction. The expression of SLPI was examined with Western blot. The levels of interleukin (IL)-8 and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay (ELISA). Goblet cell hyperplasia of lung pathological section was observed on. RESULTS: Compared with the negative control group, the expression of SLPI increased significantly after the administration of SLPI-transfected BMSCs (0.79 ± 0.06 vs 0.24 ± 0.02, P < 0.05). The levels of IL-8 and TNF-α in BMSCs and SLPI-transfected BMSCs group were lower than those in the COPD model group. Compared with the negative control group, the administration of SLPI-transfected BMSCs resulted in a further decrease in IL-8 and TNF-α in bronchoalveolar lavage fluid [(17.6 ± 1.7) vs (36.6 ± 4.0) ng/L, P < 0.05]. SLPI-transfected BMSCs transplantation also significantly attenuated goblet cell hyperplasia in rats (P < 0.05). CONCLUSION: There is a potential role for cell-based SLPI gene therapy in the treatment of COPD.