Inhibition of the Hedgehog Signaling Pathway Depresses the Cigarette Smoke-Induced Malignant Transformation of 16HBE Cells on a Microfluidic Chip.

BACKGROUND: The hedgehog signaling system (HHS) plays an important role in the regulation of cell proliferation and differentiation during the embryonic phases. However, little is known about the involvement of HHS in the malignant transformation of cells. This study aimed to detect the role of HHS in the malignant transformation of human bronchial epithelial (16HBE) cells. METHODS: In this study, two microfluidic chips were designed to investigate cigarette smoke extract (CSE)-induced malignant transformation of cells. Chip A contained a concentration gradient generator, while chip B had four cell chambers with a central channel. The 16HBE cells cultured in chip A were used to determine the optimal concentration of CSE for inducing malignant transformation. The 16HBE cells in chip B were cultured with 12.25% CSE (Group A), 12.25% CSE + 5 µmol/L cyclopamine (Group B), or normal complete medium as control for 8 months (Group C), to establish the in vitro lung inflammatory-cancer transformation model. The transformed cells were inoculated into 20 nude mice as cells alone (Group 1) or cells with cyclopamine (Group 2) for tumorigenesis testing. Expression of HHS proteins was detected by Western blot. Data were expressed as mean ± standard deviation. The t-test was used for paired samples, and the difference among groups was analyzed using a one-way analysis of variance. RESULTS: The optimal concentration of CSE was 12.25%. Expression of HHS proteins increased during the process of malignant transformation (Group B vs. Group A, F = 7.65, P < 0.05). After CSE exposure for 8 months, there were significant changes in cellular morphology, which allowed the transformed cells to grow into tumors in 40 days after being inoculated into nude mice. Cyclopamine could effectively depress the expression of HHS proteins (Group C vs. Group B, F = 6.47, P < 0.05) and prevent tumor growth in nude mice (Group 2 vs. Group 1, t = 31.59, P < 0.01). CONCLUSIONS: The activity of HHS is upregulated during the CSE-induced malignant transformation of 16HBE cells. Cyclopamine can effectively depress expression of HHS proteins in vitro and prevent tumor growth of the transformed cells in vivo.

[Effects of insulin-like growth factor-1 and insulin on modulating glucose metabolism and functions of lung fibroblasts].

OBJECTIVE: It is recognized that lung fibroblasts (LF) act as a common pathway in the development of fibrosis from alveolitis of whatever etiology. The study was undertaken to identify the modulating effects of insulin-like growth factor-1 (IGF-1) on LF glucose metabolism and functions, to explore possible therapeutic targets through modulating LF for the treatment of pulmonary fibrosis. METHODS: Human embryonic lung (HEL) diploid fibroblast cells were cultured for 24 h with blank control, 100 ng/ml IGF-1, 200 ng/ml IGF-1, 100 ng/ml IGF-1 + 100 ng/ml insulin, 100 ng/ml IGF-1 + 200 ng/ml insulin, 100ng/ml insulin, 200 ng/ml insulin, respectively. The culture supernatants and cell pellets were collected to extract proteins and mRNAs. After quantitated with ultraviolet spectrometer, RT-PCR and Northern blotting were performed to determine the content of glucose transporter-4 (Glut-4), hexokinase II, elastin and collagen-IV. RESULTS: RT-PCR showed that the general mean absorbance ratios of Glut-4 and HK-IImRNA in IGF-1 group were (0.67 +/- 0.25) and (0.60 +/- 0.19), significantly increased as compared with that of control group [(0.61 +/- 0.12), (0.55 +/- 0.19)]; but decreased as compared with that of insulin group [(0.74 +/- 0.26), (0.71 +/- 0.23)]. The expression of elastin, collagen protein in IGF-1 group [(174.3 +/- 4.2), (142.1 +/- 1.0)] were significantly stronger than that of insulin group. CONCLUSIONS: IGF-1 can stimulate LF glucose metabolism, companied with increased expression of elastin and collagen IV in a concentration-dependent manner. IGF-1 can decrease the effect of insulin on LF glucose metabolism, whereas insulin can attenuate the effect of IGF-1 on LF in elastin and collagen expression.

Z-palatoplasty and tongue radiofrequency for patients with small tonsils.

OBJECTIVE: To explore the feasibility and efficacy of Z-palatoplasty for the management of severe obstructive sleep apnea-hypopnea syndrome in patients with tonsils. STUDY DESIGN: Case series and chart review. SETTING: University hospital. METHODS: Z-palatoplasty and coblation channeling of the tongue were performed in 36 patients with body mass index <40 kg/m(2) and size 1 or 2 tonsils. All patients' tonsils were preserved. Follow-up continued for at least 1 year. Success was defined as a postoperative apnea-hypopnea index <15 events per hour and at least 50% less than the preoperative value. RESULTS: The surgical success rate was 58.3% (21/36 patients). Furthermore, 66.7% (24/36 patients) had a ≥ 50% reduction in the apnea-hypopnea index to less than 20 episodes per hour. There were statistically significant differences in preoperative nadir oxygen saturation, percentage of time with oxygen saturation less than 90%, microarousal index, and Friedman tongue position between those who responded to surgery and those who did not. Six patients had temporary velopharyngeal insufficiency. After 3 months, all the patients had normal deglutition. No major perioperative complications occurred. CONCLUSION: Our findings suggest that Z-palatoplasty can improve disease-specific quality of life and sleep apnea symptoms in patients with obstructive sleep apnea-hypopnea syndrome and size 1 or 2 tonsils.

[Research on the relationship between obstructive sleep apnea hypopnea syndrome and gastroesophageal reflux].

OBJECTIVE: To study the relationship between obstructive sleep apnea and nocturnal gastroesophageal reflux (GER). METHODS: Seventy-six patients with obstructive sleep apnea hypopnea syndrome (OSAHS) underwent polysomnography (PSG) and nocturnal distal esophageal pH monitoring, to assess the prevalence of GER in OSAHS patients. Among these patients, thirty-two OSAHS patients with GER disease had been operated on. Surgical treatment included uvulopalatopharyngoplasty (UPPP), inferior turbinate reduction, nasal septoplasty et al. After 6 months of surgical treatment, the PSG and pH probe testing were rechecked. The pre and post operative apnea-hypopnea index (AHI), micro-arousal index (MAI), lowest artery oxygen saturation (LSaO2), acidified index of distal esophagus (pHI) and percentage of acid contact time of esophagus were compared. The correlation analysis was also employed. RESULTS: The AHI of 76 OSAHS patients was (38.6 +/- 29.5)/h (average +/- s), GER was present in 48 patients (63.2 %). The pre and post operative AHI, MAI, LSaO2, pHI and percentage of ACT in 32 OSAHS patients associated with GER were (51.2 +/- 23.1)/h and (17.3 +/- 10.3)/h, (38.3 +/- 21.4 )/h and (14.5 +/- 10.0)/h, 0.698 +/- 0.107 and 0.858 +/- 0.076, (12.5 +/- 6.6)/h and (6.3 +/- 4.2)/h, (10.3 +/- 5.2)% and (4.5 +/- 2.9)%. The differences were statistically significant by matching t test (t were 10.95, 7.82, 15.97, 6.12, 7.62 respectively, P < 0.001). There were positive relationships between the reductions of AHI, MAI and the reductions of pHI (r were 0.775, 0.764, P < 0.001). The improved levels of AHI and MAI were correlated with the reduction of the percentage of ACT (r were 0.607, 0.730, P < 0.001). CONCLUSIONS: GER is prevalent in OSAHS patients. Surgical treatment of OSAHS has significant reduction in AHI, MAI as well as marked improvement in the GER, which suggests that nocturnal gastroesophageal reflux is correlated with the disorder of sleep respiration and the sleep disturbances.