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Objective The FMS-related tyrosine kinase 3 ( FLT-3 ) gene is a hematopoietic growth factor receptor gene, an independent negative prognostic factor, which affects the proliferation and differentiation of stem cells or hematopoietic progenitor cells. Patients with FLT-3 gene mutations have a worse prognosis and responsiveness to chemotherapy than those without these mutations. Our study aims to establish a conventional detection method for FLT3-ITD and D835 mutations in patients with acute myeloid leukemia (AML). Materials and methods In this study, we recruited 100 patients with AML. Primers were designed to distinguish between wild-type FLT-3 , FLT3-ITD , and D835 variants. Methods using a polymerase chain reaction (PCR)-Agilent 2100 Bioanalyzer, PCR-ABI PRISM 3100 Genetic Analyzer, and PCR–agarose gel electrophoresis were compared. Results A high-accuracy, easily operated, low-cost technique to detect the FLT-3 variation with 99.9% specificity was established in this study. The PCR platform, the Agilent 2100 Bioanalyzer (plus DNA 1000 LabChip kit) chip analysis platform, and the ABI PRISM 3100 Genetic Analyzer (plus GeneScan-500 size standard) short tandem repeat (STR) fluorescence analysis platform were used in different experimental comparisons. The ABI PRISM 3100 Genetic Analyzer (plus GeneScan-500 size standard) STR fluorescence analysis platform was the most suitable method to detect FLT3 variants. This method has a high degree of sensitivity, accuracy, and a specificity of 99.9%. Conclusion AML with the homozygous mutated FLT-3 may have a worse cure rate than AML with heterozygous mutation. This mutation is not related to drug resistance, but is a factor in a high risk of relapse; it is also related to unfavorable overall survival. Our designed detection methods should provide key information to develop personalized medicine for AML patients.

Background To identify a novel therapeutic agent for hepatocellular carcinoma (HCC), for which no promising therapeutic agent exists, we screened a panel of plants and found that Juniperus chinensis exhibited potential antiangiogenic and anti-HCC activities. We further investigated the antiangiogenic and anti-HCC effects of the active ingredient of J. chinensis extract, CBT-143-S-F6F7, both in vitro and in vivo. Methods A tube formation assay conducted using human umbilical vein endothelial cells (HUVECs) was first performed to identify the active ingredient of CBT-143-S-F6F7. A series of angiogenesis studies, including HUVEC migration, Matrigel plug, and chorioallantoic membrane (CAM) assays, were then performed to confirm the effects of CBT-143-S-F6F7 on angiogenesis. The effects of CBT-143-S-F6F7 on tumor growth were investigated using a subcutaneous and orthotopic mouse model of HCC. In vitro studies were performed to investigate the effects of CBT-143-S-F6F7 on the cell cycle and apoptosis in HCC cells. Moreover, protein arrays for angiogenesis and apoptosis were used to discover biomarkers that may be influenced by CBT-143-S-F6F7. Finally, nuclear magnetic resonance analysis was conducted to identify the compounds of CBT-143-S-F6F7. Results CBT-143-S-F6F7 showed significantly antiangiogenic activity in various assays, including HUVEC tube formation and migration, CAM, and Matrigel plug assays. In in vivo studies, gavage with CBT-143-S-F6F7 significantly repressed subcutaneous Huh7 tumor growth in severe combined immunodeficient (SCID) mice, and prolonged the survival of orthotopic Huh7 tumor-bearing SCID mice (a 40 % increase in median survival duration compared with the vehicle-treated mice). Immunohistochemical staining of subcutaneous Huh7 tumors in CBT-143-S-F6F7-treated mice showed a significantly decrease in the cell cycle regulatory protein cyclin D1, cellular proliferation marker Ki-67, and endothelial marker CD31. CBT-143-S-F6F7 caused arrest of the G2/M phase and induced Huh7 cell apoptosis, possibly contributing to the inhibition of HCC tumors. Protein array analysis revealed that several angiogenic and antiapoptotic factors were suppressed in CBT-143-S-F6F7-treated Huh7 cells. Finally, five compounds from CBT-143-S-F6F7 were identified. Conclusions According to these results, we report for the first time the antiangiogenic and anti-HCC activities of CBT-143-S-F6F7, the active fractional extract of J. chinensis. We believe that CBT-143-S-F6F7 warrants further evaluation as a new anti-HCC drug. Electronic supplementary material The online version of this article (doi:10.1186/s12906-016-1250-6) contains supplementary material, which is available to authorized users.

Angiogenesis and apoptosis array analysis of CBT-143-S-F6F7-treated Huh7 cells. (PPT 1587 kb)