Carcinoembryonic antigen in pleural effusion of patients with lung adenocarcinoma: a predictive marker for EGFR mutation.

BACKGROUND: Carcinoembryonic antigen (CEA) can reflect tumor growth, recurrence and metastasis, and also predict the clinical efficacy of the epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKI). In the present study, we investigated the association between CEA in serum and pleural effusion (PE) and EGFR mutations in patients with lung adenocarcinoma. METHODS: We retrospectively investigated 114 lung adenocarcinoma patients with malignant pleural effusion (MPE). CEA levels in serum and MPE were measured by immunoradiometric assay, we analysed the correlation between CEA and EGFR mutation status. RESULTS: Fifty-three cases had EGFR mutation (46.5%). EGFR mutations were more common in females, patients with high levels of PE (≥107.2 ng/mL) and serum CEA (≥87 ng/mL). There was no significant difference in EGFR mutation rate between in tumor tissue and PE samples (49.3% vs. 41.9%, P=0.440). The result of receiver operating characteristic (ROC) indicated that the cut off value of CEA in MPE was 107.2 ng/mL, which had the highest sensitivity (SEN) and specificity (SPE) for predicting EGFR mutation [SEN 66%, and SPE 62.3%, AUC =0.668, 95% confidence interval (CI): 0.569-0.767, P=0.025]. The combination of gender, smoking history, serum and MPE CEA level had a higher calculated AUC (0.718, 95% CI: 0.622-0.813, P=0.000). Moreover, multivariate analysis showed that CEA level in MPE but not in serum was confirmed as the only independent factor associated with EGFR gene mutation status (P=0.026) with an odds ratio of 2.885 (95% CI: 1.137-7.317). CONCLUSIONS: MPE CEA can probably serve as a predictive marker for EGFR mutation in advanced lung adenocarcinoma. Combining gender, smoking history, and CEA has a relatively better predictive value. However, detecting EGFR mutations in lung adenocarcinomas is necessary for determining EGFR-TKI treatment in clinic.

Aberrant Peripheral Immune Function in a Good Syndrome Patient.

Good's syndrome (GS) is often accompanied by recurrent respiratory infections and chronic diarrhea. The main purpose was to evaluate the peripheral immune status of a GS patient after thymoma resection. Twenty healthy volunteers were recruited as healthy controls (HCs). Flow cytometry was applied to determine the proportions of circuiting CD4+ T cells, CD8+ T cells, γδT cells, and regulatory T (Treg) cells in our GS patient. We also examined the proliferation capability of ex vivo CD4+ T cells and detected the levels of cytokines interferon- (IFN-) γ and interleukin-17A secreted by ex vivo immune cells from this GS patient. Compared with healthy control subjects, this GS patient had fewer B cells, an inverted ratio of CD4+/CD8+ cells, and more Treg cells in his peripheral blood. Additionally, the patient's Vδ2 T cell levels were significantly decreased despite having a normal percentage of γδT cells. Ex vivo peripheral CD4+ T cells from the patient showed insufficient proliferation and division potential as well as excessive expression of PD-1. Moreover, IFN-γ was predominantly derived from CD8+ T cells in this GS patient, rather than from CD4+ T cells and γδT cells. This GS patient had impaired T and B cell immunological alternations and cytokine disruptions after thymectomy. Detailed research should focus on therapies that can adjust the immune status in such patients for a better outcome.

Glomus tumors of the trachea: 2 case reports and a review of the literature.

Glomus tumors (GTs) of the trachea are very rare neoplasms that usually arise from the distal portion of the respiratory tree. The origin of these tumors is modified smooth muscle cells of glomus bodies. In this study, we describe two cases of GT of the trachea, as well as the histologic features of these tumors and their treatments. One tumor was diagnosed via bronchoscopic biopsy, and the other tumor was diagnosed via surgery. Clinical follow-up showed that the two patients are alive and well after 8 and 15 months post-treatment, respectively. We also review the literature regarding GTs and discuss the clinical presentation, histologic features, differential diagnosis, treatment and prognosis of these tumors.

Anaplastic lymphoma kinase-positive lung adenocarcinoma patient with development of sick sinus syndrome while on targeted treatment with crizotinib.

The anaplastic lymphoma kinase (ALK) positive non-small cell lung cancer (NSCLC) patients are younger and have never smoked, while pathologically are predominately adenocarcinomas. Crizotinib as an ALK inhibitor has been used in treating ALK positive NSCLC patients for several years and some adverse effects should be paid attention to. We now describe a case of ALK positive NSCLC patient with development of sick sinus syndrome (SSS) while on targeted treatment with crizotinib. A 46-year-old non-smoking woman with right hilar mass and underwent transesophageal endoscopic ultrasonography lymph node biopsy showed low differentiation adenocarcinoma, immunohistochemistry (IHC) of tumor samples revealed the ALK overexpression. The severe sinus bradycardia and RR interval prolongation were detected 3 months after crizotinib treatment, she underwent pacemaker implantation. Although the severe sinus bradycardia and RR interval prolongation were unusual adverse effects, physicians should be aware of these side effects when using crizotinib.

[The effects of imiquimod on an animal model of asthma].

OBJECTIVE: To study the mechanism of imiquimod on asthma animals. METHODS: (1) 40 mice and 48 rats were divided into 4 groups: control, asthma, dexamethasone and imiquimod groups. The asthma model was established. The mice and rats in the imiquimod group were exposed to an aerosol of 0.15% imiquimod. Lung inflammation and airway responsiveness were measured 24 h after the last ovalbumin (OVA) challenge. The expression of Interleukin-4 (IL-4), interferon gamma (IFN-gamma), eotaxin, macrophage-derived chemokine (MDC), thymus and activation-regulated chemokine (TARC), T-bet, GATA-3, STAT6 mRNA in the lung were determined by reverse transcription polymerase chain reaction (RT-PCR). The levels of eotaxin, MDC, and TARC in sera were tested by enzyme linked immunosorbent assay (ELISA). The expression of T-bet, GATA-3 and STAT6 proteins in the lung were measured by Western blot. (2) Parabronchial lymphnodes (PBLN) were isolated and cultured. The PBLN cells were divided into blank control, positive control, dexamethasone and drug groups (1 - 3 subgroups), cultured for different hours, and the expressions of IL-4 and IFN-gamma in supernatants were determined by ELISA, The mRNA expressions of the cytokines in cells weredetected by RT-PCR. (3) Flow cytometry was used to detect intracellular IL-4 and IFN-gamma production in spleen T lymphocytes. (4) CD(4)(+) T cell of spleen pellets were subject to assessment of T-bet and GATA-3 protein and mRNA expression respectively. RESULTS: The expiration resistance was determined before and after injection of acetylcholine chloride (20 - 160 microg/ml), and expiration resistances of the asthmatic group (6.26 +/- 0.85), (11.55 +/- 3.09), (28.74 +/- 5.94), (3710.83 +/- 197.49) cm H(2)Oxml(-1)xs(-1), were significantly elevated compared with those of the control group (1.34 +/- 0.16), (3.47 +/- 0.49), (9.29 +/- 1.27), (25.22 +/- 5.44) cm H(2)Oxml(-1)xs(-1), D = 88.98, 56.00, 45.00, 108.00, all P < 0.01). The numbers of eosinophils and lymphocytes, the thicknesses of WA/Pi and ASM/Pi in the asthmatic group [(26.0 +/- 1.6)/mm(2), (45.2 +/- 3.2)/mm(2), 12.0 +/- 1.4, 6.7 +/- 0.6] were all significantly higher than those of the imiquimod group [(12.4 +/- 2.9)/mm(2), (24.2 +/- 3.7)/mm(2), 9.2 +/- 0.6, 4.0 +/- 0.5, D or q = 193.00, 16.92, 185.50, 7.66, all P < 0.01]. In the imiquimod group, the mRNA and protein expressions of T-bet (0.48 +/- 0.08, 0.48 +/- 0.17) were significantly increased compared with those of the asthmatic group (0.08 +/- 0.12, 0.18 +/- 0.06, D = 120.96, 177.98, all P < 0.01), the mRNA and protein expressions of GATA-3 in the imiquimod group were both significantly decreased compared with those of the asthmatic group (D = 166.96, 310.97, all P < 0.01). In the control group, only low concentrations of IFN-gamma [(22 +/- 5, 31 +/- 5) pg/ml] were detected in PBLN cell cultures. After 24 or 48 h stimulation, the concentrations of IFN-gamma in drug 2 subgroup [(149 +/- 31), (154 +/- 28) pg/ml] and drug 3 subgroup [(166 +/- 30), (158 +/- 31) pg/ml] were increased significantly; Levels of IL-4 [druug 2 subgroup: (23 +/- 5), (39 +/- 11) pg/ml, drug 3 subgroup: (43 +/- 13), (56 +/- 12) pg/ml] were increased slowly compared with those in the OVA group (drug 2 subgroup 24 h IL-4, D = 9.90; drug 3 subgroup 24 h IL-4, D = 8.79, drug 2 subgroup 48 h IL-4, D = 8.80, drug 3 subgroup 48 h IL-4, D = 8.10, drug 2 subgroup 24 h IFN-gamma, q = 4.80, drug 3 subgroup 24 h IFN-gamma, q = 6.40, drug 2 subgroup 48 h IFN-gamma, q = 3.95, drug 3 subgroup 48 h IFN-gamma, q = 4.31, all P < 0.05). After imiquimod treatment, the mRNA and protein levels of T-bet in imiquimod group CD(4)(+) T cells were increased significantly compared with those in OVA group, and the mRNA and protein levels of GATA-3 were decreased significantly in CD(4)(+) T cells of imiquimod group compared with those in OVA group. The eotaxin, MDC and TARC levels of serum in asthma group [(593 +/- 41) pg/ml, (170 +/- 20) pg/ml, (221 +/- 25) pg/ml] were significant different from those in control group [(288 +/- 66) pg/ml, (100 +/- 33) pg/ml, (84 +/- 49) pg/ml], (eotaxin: q = 12.20, MDC: q = 8.00, TARC: q = 10.50, all P < 0.01). MDC and TARC levels of serum in imiquimod group [(84 +/- 13) pg/ml, (163 +/- 35) pg/ml] decreased as compared with those in asthma group (MDC: q = 9.80, TARC: q = 4.50, all P < 0.01) and MDC levels in imiquimod group were no different with normal group (q = 1.80, P > 0.05). eotaxin levels of serum in imiquimod group [(501 +/- 76) pg/ml] increased as compared with those from normal group (q = 8.50, P < 0.01), and decreased as compared with those from asthma group (q = 3.70, P < 0.05). (4) The expression of eoaxin, MDC, TARC and STAT(6) on the bronchial epithelium in imiquimod group was decreased as compared with asthma group, but increased as compared with normal group. The eotaxin, MDC and TARC mRNA expression of the lung in asthma group (0.85 +/- 0.11, 0.96 +/- 0.10, 0.94 +/- 0.28) had significant differences from those in the control group (0.45 +/- 0.08, 0.39 +/- 0.09, 0.24 +/- 0.08, eotaxin: q = 3.00, MDC: q = 15.40, TARC: q = 5.90, all P < 0.01) and those in imiquimod group (0.65 +/- 0.17, 0.66 +/- 0.12, 0.66 +/- 0.34, eotaxin: q = 1.50, MDC: q = 8.10, TARC: q = 2.40, all P < 0.05). CONCLUSION: These findings suggested that imiquimod can inhibit the airway inflammation of asthma animals by reducing GATA-3 mRNA and protein expression and increasing T-bet, STAT(6) mRNA and protein expression.

[Imbalanced T cell-specific transcription factors T-bet and GATA-3 and relationship to airway inflammation in asthmatic rats].

OBJECTIVE: To identify the change of the mRNA and protein expression of T-bet and GATA-3 in lung tissues, and to investigate the association between the imbalanced T cell-specific transcription factors T-bet/GATA-3 and the airway inflammation in asthmatic rats. METHODS: Twenty-four male SD rats were randomly divided into a control group and an asthmatic group. Airway responsiveness was measured and the change of airway histology was observed. The concentrations of interleukin-4 (IL-4), IL-5, and interferon-gamma (IFN-gamma) in bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expressions of IL-4, IL-5, IFN-gamma, T-bet and GATA-3 in the lungs were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively. RESULTS: The expiration resistance after injection of acetylcholine chloride (20, 40, 80, 160 microg/ml) in the asthmatic group was (6.26 +/- 0.85), (11.55 +/- 3.09), (28.74 +/- 5.94), (3,710.83 +/- 197.49) cm H2O.ml(-1).s(-1) respectively; and that in the control group was (1.51 +/- 0.18), (2.15 +/- 0.36), (6.08 +/- 1.06), (37.17 +/- 6.12) cm H2O.ml(-1).s(-1) respectively; the difference being significant between the two groups (all P < 0.01). In the asthmatic group, the numbers of eosinophils and lymphocytes, the thicknesses of WA/Pi and ASM/Pi were (26.0 +/- 1.6)/mm(2), (45.2 +/- 3.2)/mm(2), 12.0 +/- 1.4, 6.7 +/- 0.6, respectively; and those of the control group were (2.9 +/- 1.2)/mm(2), (8.8 +/- 1.8)/mm(2), 6.4 +/- 0.8, 2.7 +/- 0.5, respectively; all were significantly different between the two groups (all P < 0.01). In the asthmatic group, the concentrations of IL-4, IL-5, and IFN-gamma in BALF were (23.4 +/- 0.7) pg/ml, (24.8 +/- 0.5) pg/ml, (21.7 +/- 1.1) pg/ml, respectively, and those of the control group were (9.3 +/- 0.3) pg/ml, (12.5 +/- 0.3) pg/ml, (65.8 +/- 2.1) pg/ml, respectively; all were significantly different between the two groups (all P < 0.01). In the control group, the mRNA ratio of T-bet to GATA-3 (0.73 +/- 0.32) was significantly increased compared with the asthmatic group (0.06 +/- 0.09, P < 0.01). There was also a significant difference in the ratio of protein expression of T-bet to GATA-3 between the control group (0.75 +/- 0.25) and the asthmatic group (0.09 +/- 0.04, P < 0.01). The ratio of protein expression of T-bet and GATA-3 was correlated negatively with expiration resistance (r = -0.959, -0.919, -0.949, all P < 0.01), the numbers of eosinophils and lymphocytes in lung tissues (r = -0.832, -0.831, all P < 0.01), the thicknesses of WA/Pi and ASM/Pi (r = -0.837, -0.863, all P < 0.01) and the concentrations of IL-4, IL-5 in BALF (r = 0.921, 0.920, all P < 0.01), the mRNA of IL-4, IL-5 in lung tissues (r = -0.964, -0.931, all P < 0.01), but positively with the concentrations of IFN-gamma in BALF and the mRNA of IFN-gamma in lung tissues (r = -0.934, 0.983, all P < 0.01). CONCLUSION: Imbalance of transcription factors T-bet and GATA-3, a reflection of the immune imbalance in asthma, may play a key role in the formation of airway inflammation in the disease.

Treatment of allergic airway inflammation and hyperresponsiveness by imiquimod modulating transcription factors T-bet and GATA-3.

BACKGROUND: Imiquimod is an imidazoquinoline, which class of compounds are known to have antiviral and antitumoural properties. In recent studies, it was shown that imiquimod modulates the T helper cell type Th1/Th2 response by inducing the production of Th1 cytokines like IFN-gamma, and by inhibiting the Th2 cytokines like interleukin (IL)-4. Several investigators have shown that T-bet and GATA-3 are master Th1 and Th2 regulatory transcription factors. This study investigated whether imiquimod treatment inhibited airway inflammation by modulating transcription factors T-bet and GATA-3. METHODS: Thirty-six male SD rats were randomly divided into a control group, an asthmatic group, and an imiquimod group, which was exposed to an aerosol of 0.15% imiquimod. Twenty-four hours after the last ovalbumin (OVA) challenge, airway responsiveness was measured and changes in airway histology were observed. The concentrations of IL-4, IL-5 and IFN-gamma in bronchoalveolar lavage fluid (BALF) and serum were measured by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of IL-4, IL-5, IFN-gamma, T-bet and GATA-3 in lung and in CD4(+) T cells were determined by reverse transcription polymerase chain reaction (RT-PCR). The protein expressions of T-bet and GATA-3 were measured by Western blot. RESULTS: It was demonstrated that imiquimod 1) attenuated OVA induced airway inflammation; 2) diminished the degree of airway hyperresponsiveness (AHR); 3) decreased the Th2 type cytokines and increased Th1 type cytokines mRNA and protein levels; 4) modulated the Th1/Th2 reaction by inhibiting GATA-3 production and increasing T-bet production. CONCLUSION: Imiquimod treatment inhibits OVA induced airway inflammation by modulating key master switches GATA-3 and T-bet that result in committing T helper cells to a Th1 phenotype.