The amino acid and carnitine concentration changes in bronchoalveolar lavage fluid from lung cancer patients.

OBJECTIVE: To screen out potential biomarkers by analyzing fundamental nutrients in the bronchoalveolar lavage fluid (BALF) before confirming the lung cancer. METHODS: In this study, 44 patients were enrolled with clinical information. The concentrations of 23 amino acids and 35 carnitines in their BALF were detected with the high-performance liquid chromatography-mass spectrometry (HPLC-MS). Combined with clinicopathological diagnosis, the patients were divided into the lung cancer group (grades I & II and III & IV) and the non-cancer group for standard statistical analysis. RESULTS: The partial least squares-discriminant analysis (PLS-DA), the Shapiro-Wilk test, and the Bonferroni correction results showed that the serine concentration was higher and the butane-diacyl-carnitine (C4DC) concentration was lower in the lung cancer group, further showing the same changing trend continuously through the non-cancer stage, grades I & II stage and grades III & IV stage. Those two potential biomarkers have been identified. CONCLUSION: The HPLC-MS target detection in clinic for nutrient concentration levels is a promising technique to find the changing concentration of serine and C4DC in BALF, which provides an economical and practical way for early warning of lung cancer.

Graphitic carbon nitride quantum dots as analytical probe for viewing sialic acid on the surface of cells and tissues.

The abnormal expression of sialic acids (SAs) on cells and tissues is closely related to various pathophysiological states. Here we applied phenylboronic acid (PBA) functionalized graphitic carbon nitride fluorescent quantum dots (PCQDs) with sizes from 3 to 5 nm in efficient and selective labeling SAs on the surface of living cells and tissues. With abundant PBA in their structure, the water soluble PCQDs showed the relative SA level on the cell surface via selectively and efficiently staining different cell lines in 30 min and revealed that M1 macrophages may express more SAs on their surfaces compared with M0 and M2. The distinct demarcation of cancerous and para-noncancerous areas on cancer tissue sections was showed by PCQDs staining. PCQDs with their high selectivity, stable photoluminescence, low cost, and nontoxicity can be an ideal SA fluorescent probe for living cells and tissues.

In situ detecting changes in membrane lipid phenotypes of macrophages cultured in different cancer microenvironments using mass spectrometry.

Macrophages, the important cells of immune system, have exhibited distinct gene phenotypes with diverse functions in different microenvironments. In the present study, macrophages RAW264.7 (M0 macrophages) and lipopolysaccharide (LPS) plus interferon gamma (INF-γ)-treated M0 macrophages (M1 macrophages) were cultured in different lung cell-derived culture supernatants (CSs) as imitative tumor microenvironments. The lipids (mainly from cell membrane) of intact macrophages were in situ detected by matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry. Approximately 300 of small molecules were observed in negative ion mode. Partial least square-discriminant analysis (PLS-DA) suggested that two types of the macrophages have different membrane lipid phenotypes. Changes in the levels of phosphatidylethanolamine PE(16:1/18:0), PE(18:1/18:0), PE(36:2), PE-Cer(d36:1), and PE(P-16:0/18:1) were closely associated with membrane phenotypes of macrophages. The heatmap also revealed that directional induction to classically activated macrophages (M1 macrophages) in vitro had greater impact on the membrane lipid phenotypes of macrophages than different lung cell-derived CSs. The results are consistent with the data obtained by biological technologies.

Simultaneous Quantification of Serum Nonesterified and Esterified Fatty Acids as Potential Biomarkers to Differentiate Benign Lung Diseases from Lung Cancer.

In this study, we have employed graphene oxide as a matrix to simultaneously and directly quantify serum nonesterified and esterified fatty acids (FAs) using matrix-assisted laser/desorption ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS). Twelve serum nonesterified FAs combined with their individual esterified FAs (i.e., C16:0, C16:1, C18:0, C18:1, C18:2, C18:3, C20:2, C20:3, C20:4, C20:5, C22:5, and C22:6) were quantified based on their calibration curves with the correlation coefficients of >0.99, along with the analytical time of <1 min each sample. As a result, serum levels of twelve total FAs (TFAs) in 1440 serum samples from 487 healthy controls (HCs), 479 patients with benign lung diseases (BLDs) and 474 patients with lung cancer (LC) were determined. Statistical analysis indicated that significantly increased levels of C16:0, C16:1, C18:0, C18:1, C18:3, C20:3, and C22:6 and decreased levels of C20:5 were observed in LC patients compared with BLDs. Receiver operating characteristic (ROC) analysis revealed that panel a (C18:2, C20:3, C20:4, C20:5, C22:5, and C22:6), panel b (C18:0, C20:4, C20:5, and C22:6), and panel c (C16:1, C18:0, C18:1, C20:3, and C22:6) have exhibited good diagnostic ability to differentiate BLDs from LC relative to clinical uses of tumor markers (CEA and Cyfra 21-1).