[Effect of Wnt signaling suppression on gefitinib in non small cell lung cancer cell lines].

OBJECTIVE: To explore the effect of Wnt signaling suppression on proliferation of non small cell lung cancer to gefitinib, and its related mechanisms. METHODS: PC9 and PC9/AB2 cells of both gefitinib sensitive and resistant were treated with different concentrations of gefitinib, and the proliferation index was measured using CCK8 kit. The members of Wnt signaling pathway were detected by Western blot. Dual luciferase reportor gene assay (TOP Flash) was used to document the transcriptional level of ß-catenin. ß-catenin siRNA was transfected into PC9/AB2 cells to suppress the Wnt signaling transcription, followed by treatment with different concentrations of gefitinib. Western blot was then used to detect the expression of EGFR and its downstream signaling after inhibit the expression of ß-catenin. RESULTS: Treating with different concentrations of gefitinib, the resistance of PC9/AB2 cells to gefitinib was significantly increased (P < 0.05). The members of Wnt signaling expressed at higher level in PC9/AB2 cells than in PC9 cells (t = 24.590, P = 0.000). TOP Flash examination showed that the endogenous transcriptional activity of Wnt signaling was higher in PC9/AB2 cell than that in PC9 cell (t = 4.983, P = 0.008). Compared with the negative control group, apoptotic rate and sensitivity to gefitinib significantly increased in interfered group (P < 0.05). The expression of p-ERK1/2 significantly decreased after Wnt signaling suppression, although other proteins showed no significant alterations. CONCLUSION: Suppressing the activity of Wnt signaling can partly reverse the celluar resistance to gefitinib in non small cell lung cancer.

[The ultrastructural features of sputum deposition and its value in the diagnosis of pulmonary alveolar proteinosis].

OBJECTIVE: To investigate the ultrastructural features of sputum deposition (SD) and its value in the diagnosis of pulmonary alveolar proteinosis (PAP). METHODS: Seven patients with PAP diagnosed by lung biopsy and cytology were enrolled in this study. The patients consisted of 5 men and 2 women, whose median age was 48 years (range 36 to 73). SD and bronchoalveolar lavage fluid (BALF) sediment were made into ultrathin sections and observed under transmission electron microscope (TEM), respectively. Seven cases of control group composed of 4 men and 3 women whose median age was 49 years (range 39 to 68) including 3 cases of bacterial pneumonia, two cases of COPD and 2 cases of exudative pulmonary tuberculosis. Each SD was made into ultrathin section, and compared with the experimental group. RESULTS: In PAP group, Periodic acid-schiff (PAS) staining was performed on 7 sputum smears and none of them was tested positive for any components with diagnostic interest. Four cases from the 7 paraffin-embed sections of BALF sediment by microscopic examination suggested PAP. Under TEM, BALF sediment showed that many lamellar bodies existed in and outside alveolar epithelial cells, and 5 specimens were consistent with PAP diagnosis. Compared with BALF sediment, SD had apparent degeneration with more myelin phagosomes in the cytoplasm of macrophages, more lamellar bodies in alveolar epithelial cells, and lots of lamellar bodies in the shape of concentric circle in the extracellular spaces. Four from the 7 SD samples were consistent with the diagnosis of PAP. No significant difference was found between SD and BALF in the diagnosis of PAP by electronic examination (P > 0.05). In the 7 cases of control group no drifting osmiophilic lamellar bodies in extracellular space were detected. CONCLUSIONS: The osmiophilic lamellar bodies with diagnostic value were found in SD and BALF of patients with PAP. TEM of SD in combination with clinical manifestations and radiologic findings can make a definitive diagnosis of PAP, especially for those patients who have contraindications to lung biopsy and lung lavage.

[A study on the efficacy of glucocorticoid therapy for idiopathic nonspecific interstitial pneumonia].

OBJECTIVE: To investigate the efficacy of glucocorticoid therapy in idiopathic nonspecific interstitial pneumonia (INSIP). METHODS: Twenty-nine cases of INSIP confirmed by clinical- radiological-pathological (CRP) diagnosis were collected and classified into 2 groups according to the degree of fibrosis: INSIP-1 (including 9 cases of cellular type) and INSIP-2 (20 cases of mixed and fibrotic type). Thirty cases of usual interstitial pneumonia (UIP) confirmed by CRP diagnosis served as the control. Clinical and pathological features, therapeutic effects of glucocorticoids and the follow-up results were retrospectively analyzed and the survival curves were evaluated by Kaplan-Meier method. RESULTS: The mean age at onset of INSIP-1 group [(48 ± 5) years] was significantly younger than INSIP-2 group [(52 ± 11) years] and the UIP group [(57 ± 14) years]. The course of disease in INSIP-1 group [(60 ± 28) months] was longer than that in INSIP-2 group [(48 ± 33) months] and that in the UIP group [(44 ± 23) months], but the differences were not statistically significant (F = 1.22, all P > 0.05). The efficacy rate of glucocorticoid treatment in INSIP-1 group (9/9) was higher than that in INSIP-2 group (11/20) and that in the UIP group (2/30), the differences being statistically significant (all P < 0.05). The follow-up period for INSIP-1 group [(56 ± 27) months] was significantly longer than for INSIP-2 group [(23 ± 18) months] and for the UIP group [(25 ± 17) months], and the rate of significant improvement (6/9) was higher than that of the INSIP-2 group (9/20) and the UIP group (0/30), the differences being statistically significant (F = 9.224, all P < 0.05). The mortality of INSIP-1 group (0/9) was lower than that in INSIP-2 group (4/20) and the UIP group (16/30), the difference being statistically significant (exact probability value 0.000 - 0.005, P < 0.05). CONCLUSIONS: The degree of fibrosis of INSIP is closely correlated with the effect of glucocorticoid therapy and prognosis. The cellular type has a favorable reaction to glucocorticoid therapy and a better prognosis as compared to the fibrotic type.