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Objective: The chemical constituents in the methanol extract from Farfarae Flos were rapidly identified using UPLC-Q-TOF-MS in positive and negative ion modes. Methods: The analysis was performed on an Agilent Poroshell 120 EC-C18 chromatographic column (100 mm × 2.1 mm, 2.7 μm). The mobile phase consisted of acetonitrile and 0.1% aq formic acid. In positive ion mode, gradient elution: 0-1 min, 5%-17% B; 1-3 min, 17%-19% B; 3-14 min, 19%-44% B; 14-16 min, 44%-66% B; 16-26 min, 66%-87% B; 26-28 min, 87%-95% B; 28-33 min, 95% B. In negative ion mode, gradient elution: 0-2 min, 5%-14% B; 2-10 min, 14%-32% B; 10-15 min, 32% B. The flow rate was 0.4 mL/min, and the injection volume was 5 μL. The information of the compounds was analyzed by positive and negative ion modes mass spectrum information, elements composition, reference substance retention time or mass spectrum parameters of compounds in literature. Results: Thirty-four compounds in Farfarae Flos extracts were identified, combined with provided accurate molecular weight compounds by UPLC-Q-TOF-MS, including 12 kinds of terpenoids, eight kinds of flavonoids, seven kinds of phenolic acids, two kinds of pyran compounds, one kind of phenolic ketones, one kind of fat ketones, and three kinds of alkaloids. Conclusion: The method provides the theory basis for quality control and the clinical reasonable application and provides the reference for clarifying its efficacy material base.
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<p><b>OBJECTIVE</b>To investigate the BRAF V600E mutation in Chinese patients with langerhans cell histiocytosis (LCH) and its clinical significance.</p><p><b>METHODS</b>The clinical records of 50 pathology-diagnosed LCH cases were analyzed retrospectively and the Paraffin-embedded tissue blocks were retrieved to test the BRAF V600E mutation by immunohistochemical method with a specific BRAF V600E protein antibody. BRAF V600E mutation was also tested by High-resolution Melting Analysis (HRM) followed by Sanger sequence in 31 cases.</p><p><b>RESULTS</b>BRAF V600E mutation was found in 58% (29/50) LCH cases by gene test or protein test. BRAF V600E expression was identified in 56% (28/50) LCH cases by immunohistochemical analysis alone while 54.8% (17/31) by HRM-Sanger alone. Two out of 14 cases, who were negative for BRAF V600E by HRM-Sanger analysis, were positive by immunohistochemical tests (14.3%). Otherwhile, only 1 out of 17 LCH cases with a positive BRAF V600E mutation by gene test were negative by immunohistochemical analysis. The status of BRAF V600E did not show significant relevance with age, sex, clinical stage or response. Also, BRAF V600E had no effect on the 3-year survival or event-free survival in this group.</p><p><b>CONCLUSION</b>Immunohistochemical tests for BRAF V600E in Paraffin-embedded tissue is of ideal sensitivity, 58% Chinese LCH patients have BRAF V600E positive and so LCH is a clonal disease but the exact role of BRAF V600E in LCH needs further research.</p>