Analysis of p16 gene mutations and their expression using exhaled breath condensate in non-small-cell lung cancer.

The aim of the present study was to investigate the mutational status of exons 1 and 2 of the p16 gene in the exhaled breath condensate (EBC) of patients with non-small-cell lung cancer (NSCLC) and determine the feasibility and clinical significance of applying EBC in the diagnosis of NSCLC. Polymerase chain reaction and DNA sequencing were applied to detect exon 1 and 2 alterations of the p16 gene in EBC by comparing 58 samples from NSCLC patients and 30 from healthy controls. Of the 58 EBC samples from NSCLC patients, 54 were successfully tested and 8 cases of mutations were identified, of which 3 were in exon 1 and 5 in exon 2. The mutation rate was 14.81% (8/54). There were no p16 gene mutations in the 30 samples obtained from healthy controls. EBC p16 gene mutations exhibited no statistically significant differences according to gender, smoking history, pathological type, degree of differentiation and presence or absence of lymph node metastasis. The p16 gene mutation rate was proportional to the tumor stage (P<0.05). Therefore, the detection of the p16 gene mutation in EBC may be used as a novel molecular marker to assist in the diagnosis of NSCLC.

Methylation of P16 in exhaled breath condensate for diagnosis of non-small cell lung cancer.

BACKGROUND: Non-small cell lung cancer is the most frequently cause of cancer-related death in the world. To explore the technical feasibility, we detected aberrant promoter methylation of P16 in exhaled breath condensate which was a new, non-invasive tool for diagnosis and screening program of NSCLC. METHODS: We analyzed aberrant promoter methylation of P16 in 180 samples from 60 individuals, including 30 NSCLC patients (cancer tissues, adjacent normal lung tissues, blood plasma, and EBC), and 30 healthy controls (blood plasma and EBC) by fluorescent quantitative methylation-specific polymerase chain reaction (F-MSP). RESULTS: The positive rate of aberrant promoter methylation of P16 was 26 of 30 (86.66%) in tumor tissues, 15 of 30 (50%) in blood plasma, and 12 of 30 (40%) in EBC, we have not observed the positive methylation of P16 in the adjacent normal lung tissues, or in EBC or blood plasma from the healthy control group. CONCLUSION: We found that detected promoter methylation of P16 in EBC was feasibility, it should be an useful biomarker for diagnosis of NSCLC, it have potential prospect that detected the gene molecular in EBC because of noninvasive, specificity, convenient and repeatable.

[Effects of xuebijing on nitric oxide and VEGF-A in exhaled breath condensate of patients with ALI/ARDS].

OBJECTIVE: To observe the clinical significance of changes of nitric oxide (NO) and vascular endothelial growth factor A (VEGF-A) in exhaled breath condensate (EBC) of patients with acute lung injury/acute respiratory distress syndrome (ALI/ARDS) after they were treated by Xuebijing (XBJ), and to evaluate the effect of the EBC detection technology. METHODS: Totally 32 ALI/ARDS patients receiving mechanical ventilation at intensive care unit (ICU) were randomly assigned to the treatment group and the control group, 16 cases in each group. Patients in the control group were treated by routine therapy, while those in the treatment group were treated by routine therapy + XBJ. The therapeutic course for all was 5 days. The EBC sample was collected by improved EcoScreen condenser within 24 h after confirmed diagnosis of ALI/ARDS and on the fifth day of medication. The levels of NO and VEGF-A were measured by EIA in EBC and serum. The changes of NO and VEGF-A in EBC were observed before and after treatment. RESULTS: Compared with before treatment, the level of NO in EBC and serum decreased and VEGF-A increased after treatment, showing statistical difference (P < 0.05, P < 0.01). After treatment the level of NO in EBC and serum was lower in the treatment group than in the control group (P < 0.05). The VEGF-A in EBC was higher in the treatment group than in the control group (P < 0.05). There was no statistical difference in the serum VEGF-A level between the two groups (P > 0.05). CONCLUSIONS: XBJ was an effective therapeutic drug capable to control the in vivo inflammation reaction in patients with ALI/ARDS. The detection of changes of VEGF-A and NO levels by EBC could judge the inflammatory reaction degree in ALI/ARDS patients, and help evaluating the therapeutic effect.

[Determination and clinical implication of leukotriene B(4) and tumor necrosis factor-alpha in condensate of exhaled breath of chronic obstructive pulmonary disease patients].

OBJECTIVE: To study the changes and clinical implication of leukotriene B(4) (LTB(4)) and tumor necrosis factor-alpha (TNF-alpha) in exhaled breath condensate (EBC) of chronic obstructive pulmonary disease (COPD) patients. METHODS: EBC of 20 patients in acute episode of COPD (AECOPD), 20 patients in period of remission of COPD, and 20 persons who were having regular check-up (healthy control group) were enrolled. The concentrations of LTB(4) and TNF-alpha in EBC were assayed. Forced expiratory volume in one second (FEV1) and peak expiratory flow (PEF) of COPD patients were observed at the same time, pH, oxygenation index (PaO(2)/FiO(2)), partial pressure of oxygen in arterial blood (PaO(2)) and leukocyte count were also determined. RESULTS: (1)The concentrations of LTB(4) in EBC of AECOPD (35.43+/-14.19)ng/L and remission of COPD(24.39+/-13.75)ng/L, were significantly higher than that of the healthy control group (16.75+/-7.44)ng/L, and the concentration of LTB(4) in EBC during remission of COPD was significantly lower than that of AECOPD (all P<0.05). (2)The concentration of TNF-alpha in EBC of AECOPD (9.35+/-8.66) ng/L was significantly higher than that of remission of COPD (4.42+/-4.11)ng/L and healthy control group (4.45+/-3.92) ng/L, and the differences had statistical significance (both P<0.05). The concentration of TNF-alpha in EBC showed no significant difference between patients in remission of COPD and healthy control group. (3)The concentration of LTB(4) in EBC had negative correlation with FEV1 of AECOPD patients. Regression equation was y=or-0.51 x+0.22, r=-0.481 (P<0.05). CONCLUSION: The concentrations of LTB4 and TNF-alpha in EBC of AECOPD patients are raised when oxidation stress is reinforced, and its level reflects the severity and prognosis of COPD.

[Development and application of the software for analysis of arterial blood gases in treatment suggestions and training].

OBJECTIVE: Suggestions and training in developing a computer software for analysis of arterial blood gases in treatment to be used in clinic and teaching. METHODS: The software was edited by means of Delphi 7.0 and closely combined with Windows operating system. The knowledge base was built with Microsoft Access and HTML format files. The treatment suggestions and knowledge base of the software were built according to the correlated diagnosis and treatment guidelines of Chinese Medical Association, Practical Medicine (edition 12) edited by CHEN Hao-zhu, Medicine (edition 6) edited by YE Ren-gao, Modern Clinical Blood Gases Analysis edited by QIAN Gui-sheng, Concise Clinical Blood Gas Analysis edited by LUO Yan-jie, and the experience of the experts of our hospital. RESULTS: (1)The model of treatment suggestions consisted of the treatment objective and the treatment suggestions,and its contents included simple acid-base disturbance, mixed acid-base disturbance, respiratory failure,metabolic disturbances of sodium potassium and chloride. The calculation formulae included calculation formulae for the correction of acid-base disturbance and electrolyte disturbance. (2)Arterial blood gas analysis training software contained knowledge database of arterial blood gas analysis, clinical opinions of arterial blood gas analysis, exam database of arterial blood gas analysis, knowledge of arterial blood gas analysis graph, calculation formulas, correlated books and websites,assistance and so on. CONCLUSION: The software could be used as a reference by clinical doctors in clinic, it could also be used for teaching and training. It is scientific, state-of-the art, and valuable in clinical practice.

[Correlation between the area of arterial blood gases graph and the acid-base disturbance in 92 cases with chronic obstructive pulmonary disease].

OBJECTIVE: To study the correlation between the area of arterial blood gases graph (ABGG) and the acid-base disturbance (ABD). METHODS: Using the software developed by ourselves, the results of arterial blood gases and concomitant electrolyte (K(+), Na(+), Cl(-), HCO(3)(-)) determination in 313 episodes were analyzed in 92 cases with chronic obstructive pulmonary diseases (COPD) during hospitalization, and the correlation was studied. RESULTS: (1)On admission, among 92 cases, in 9 cases the results of blood gas analysis were in the area of compensated ventilation and deranged gas exchange (area IV), in 82 cases the results were in the area of insufficient ventilation and deranged gas exchange (area V), and 1 case in the area of excessive ventilation and deranged gas exchange (area VI). There were 8 types of the ABDs, and respiratory acidosis (RAC)+metabolic alkalosis (MAL), RAC, RAC+metabolic acidosis (MAC) with increase in anion gap (AG) ranked in above order. (2)At the time of discharge, in 2 cases the distribution of ABGG was in the normal area (area I), 5 cases in the area IV, 20 cases in the area V, and 1 case in the area VI. There were 5 types of the ABDs. RAC+MAL and RAC ranked in the first two positions. (3)During hospitalization, the correlations between the area of ABGG and ABD in 313 episodes were as follows: ABD normal and MAC of AG increased in the area I; ABD normal and MAL, MAC with AG increased, RAC+MAC, respiratory alkalosis (RAL)+MAL, MAL+MAC of AG increased, RAC+MAL+MAC of AG increased and RAL+MAL+MAC of AG increased in the area IV; RAC, MAL, RAC+MAL, RAC+MAC of AG increased, MAL+MAC of AG increased, and RAC+MAL+MAC of AG increased in the area V; MAC of AG increased, RAC+MAC, RAL+MAL, MAL+MAC of AG increased, and RAL+MAL+MAC of AG increased in the area VI. No ABD occurred in the areas II and III. CONCLUSION: The correlation between ABGG and ABD can be analyzed easily and quickly and the accuracy of results is ensured by using the software.

[Clinical application of a software for the analysis of arterial blood gases graph in 231 patients with chronic obstructive pulmonary disease].

OBJECTIVE: To evaluate the clinical significance of a computer software for the analysis of arterial blood gases graph (ABGG) in chronic obstructive pulmonary disease(COPD). METHODS: The software was developed with Win98 as the operating platform and the visual software Delphi 5.0 from Borland Company, and it was used for evaluation of the changes in arterial blood gases (ABG) of 231 COPD cases. RESULTS: (1) With the software it took only (4.7+/-0.5)s to draw and analyze an ABGG of COPD patients during oxygen inhalation; the time was much shorter than manual analysis (90.2+/-4.9)s, P<0.001. (2) During acute attack, the distributions of the arterial blood gases parameters on ABGG were as follows: 55.4% of cases in the area of insufficient ventilation and deranged gas exchange (area 5), 22.9% of cases in the area of compensated ventilation and deranged gas exchange (area 4), 21.6% of cases in the area of excessive ventilation and deranged gas exchange (area 6). (3) When the COPD patient's condition improved, the location of ABG in ABGG shifted from area 5 to area 4 or area 6. The distributions of the arterial blood gases parameters on ABGG of 106 cases on admission were significantly different from those at the time of discharge. (4) With deterioration of patient's condition, it shifted to area 5. Before death, the arterial blood gases parameters were exclusively in the area 5. CONCLUSION: The computer software for ABGG shortened the time to draw and evaluate ABG during oxygen inhalation, and it could reflect the changes in patient's condition promptly.