Objective Comparison of three different molecular assays for the detection and molecular characterization of circulating tumor cells in breast cancer .Methods The retrospective study compared three different molecular assays to detect CTC in the peripheral blood of 30 healthy individuals and 71 benign breast disease patients and 83 early and 84 metastatic breast cancer patients .All samples were collected at the outpatient , inpatient and physical examination department of Sichuan Provincial People ′s Hospital from January 2011 to June 2014.The same cDNAs were analyzed by:singleplex RT-qPCR assay for BCL-2, multiplex RT-qPCR for BCL-2, HER-2, HMAM, and a commercially available molecular assay (AdnaTest BreastCancer ) for GA733-2, MUC-1, HER-2.The positive of CTC were compared among healthy individuals and benign breast disease patients and breast cancer patients .Chi square test was used to compare the expression of gene markers among the three groups , and the agreement of Kappa test was used to evaluate the method.Results (1) Detection rates of early breast cancer by single RT-qPCR, Adna kits and multiple RT-qPCR were 13.3%, 16.9% and 18.1%, respectively , and the detection of metastatic breast cancer were 31.0%, 42.9%and 35.7%, respectively.There were significant differences in the positive of CTC by three molecular assays between healthy individuals and benign breast disease patients and early breast cancer patients ( The test values were 4.235 and 4.301, 5.367 and 5.474, 5.894 and 6.023 respectively, P<0.05).There were no differences between benign breast disease patients and early breast cancer patients (The test values were 0.891,0.748 and 0.701 respectively,P >0.05) .There were significant differences between metastasis breast cancer patients and healthy individuals and benign breast disease patients and early breast cancer patients ( The test values were 8.429,7.553 and 7.061;10.24, 9.025 and 8.745; 9.658, 8.417 and 8.201 respectively,P<0.05).(2) In early breast cancer: The concordance between AdnaTest and single RT-qPCR was 79.5%while between AdnaTest and multiplex RT-qPCR was 77.1%.No agreement was found among them ( The test values were 1.065 and 1.871, P were 0.371 and 0.258 ) .The concordance between single RT-qPCR and multiplex RT-qPCR was 80.7%.No agreement was found between them (The test values was 2.814, P was 0.156).(3) In patients with overt metastasis:The concordance between AdnaTest and single RT-qPCR was 78.6%( The test values was 10.986).While between AdnaTest and multiplex RT-qPCR was 80.9%( The test values was 9.251 ) . Agreements were found among them ( P was 0.002 and 0.005 respectively ) .The concordance between single and multiplex RT-qPCR was 88.1%( The test values was 12.364 ) .Agreement was found between them (P was 0.001).Conclusions No correlations were found among different molecular methods to detect CTC in the early primary breast cancer , but correlations were found in the metastatic breast cancer , suggesting that different rate of CTC caused by the number of CTC and its heterogeneity should be considered to the clinical diagnosis and treatment of breast cancer while molecular method is used .

OBJECTIVE To study the effects of bergapten in the treatment of liver fibrosis and its mechanism based on serum metabolomics. METHODS Forty mice were divided into normal control group （0.5% carboxymethyl cellulose sodium solution）， model group （0.5% carboxymethyl cellulose sodium solution）， and BP low-dose and high-dose groups （50， 100 mg/kg）， with 10 mice in each group. Except for the normal control group， the other three groups were all treated with carbon tetrachloride to induce liver fibrosis model； they were given relevant medicine/solution intragastrically， once a day， for consecutive 8 weeks. After the last medication， the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum were detected， and liver pathological changes were observed； the expressions of α-smooth muscle actin （α-SMA） and Collagen Ⅰ were detected in liver tissue； the serum of the mice was collected for metabolomics analysis. RESULTS Compared with the model group， serum levels of ALT and AST and protein expressions of α-SMA and Collagen Ⅰ in liver tissue were decreased significantly in BP high-dose and low-dose groups （P＜0.05）， while liver fibrosis was improved significantly. Meanwhile， metabolomics analyses showed that there were a total of 175 serum differential metabolites in the BP high-dose group and model group， of which 18 substances were upregulated and 157 substances were downregulated； the main metabolic pathways involved in bergapten intervention were pyrimidine metabolism， butanoate metabolism， fatty acid synthesis， tyrosine metabolism， β-alanine metabolism， nicotinic acid and nicotinamide metabolism， glutathione metabolism， etc. CONCLUSIONS BP is effective in the treatment of liver fibrosis by regulating pyrimidine metabolism， butanoate metabolism， glutathione metabolism and so on in rats with liver fibrosis.

OBJECTIVE To investigate the improvement mechanism of hyperoside （HYP） on non-alcoholic fatty liver disease （NAFLD）. METHODS Male C57BL/6 mice were randomly divided into normal （NFD） group， model （HFD） group and HYP group， with 8 mice in each group. Except for NFD group， the mice in other groups were fed with HF60 high-fat diet to establish NAFLD model； HYP group was simultaneously given HYP 100 mg/kg intragastrically every day， for 16 consecutive weeks. The body weight and liver weight of mice in each group were recorded 16 h after the last medication； the histopathological changes and lipid accumulation in the liver were observed， and the contents of triglyceride （TAG） in liver tissue and serum contents of TAG， aspartate transaminase （AST） and alanine transaminase （ALT） were measured； LC-MS/MS method was adopted to detect lipid changes in the liver tissue of mice for lipidomics analysis， and protein expressions of lipid synthesis-associated proteins peroxisome proliferator-activated receptor α （PPARα） were also tested. Human hepatocellular carcinoma cell line HepG2 was divided into normal control group， model group， HYP low-concentration group （50 μmol/L）， HYP high-concentration group （100 μmol/L）， HYP low-concentration+GW6471 （PPARαinhibitor） group， and HYP high-concentration+GW6471 group. Except for normal control group， the remaining cells were induced with oleic acid and palmitic acid to establish a high-fat cell model. The accumulation of lipid droplets in each group of cells was observed， and the TAG content was detected. RESULTS Compared with HFD group， HYP group exhibited significant reductions in liver fat vacuoles， lipid accumulation， liver weight， and TAG content in liver tissue， as well as serum contents of ALT， AST and TAG （P＜0.05）. Additionally， the expression of PPARα protein in liver tissue was significantly increased （P＜0.05）， and the pathological morphological changes associated with NAFLD were alleviated. Lipidomic analysis revealed that HYP significantly reduced the levels of TAG， diacylglycerol and other lipids in the liver. Compared with model group， cellular lipid droplet accumulation and TAG content decreased significantly in HYP low- and high-concentration groups （P＜0.05）； GW6471 could significantly reverse the improvement effect of HYP on above indicators （P＜0.05）. CONCLUSIONS HYP can effectively ameliorate NAFLD induced by a high-fat diet in mice， and the mechanism may be related to the activation of PPARα to regulate hepatic lipid synthesis.