Guan Xin Shu ( GXS ) is a heparinoid.Quails were randomly divided into 2 groups. All of them were fed with inducer diet containing 1% cholesterol and 20% fat for 6 weeks. 1 group was treated with GXS (200mg/kg/d, ip ) , the other with 0.9% NaCl(0.2ml/100g/d, ip ) . 4 and 6 weeks after administration, serum cholesterol level was significantly reduced (mean 3l%, P

Objective:To study the effect of astrocyte elevated gene-1 (AEG-1) on proliferation and metastasis of papillary thyroid cancer (PTC) by regulating cell autophagy and epithelial-mesenchymal transition (EMT).Methods:Normal thyroid cells Nthy-ori3-1 and PTC cells TPC-1, FTC-133, B-CPAP and SW579 were cultured in vitro. Real time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of AEG-1 in PTC cells. The PTC cells with the highest AEG-1 expression were selected for AEG-1 shRNA infection, and then divided into sh-NC group and sh-AEG-1 group. Cell counting kit-8 (CCK8) and EdU (5-ethynyl-2 ′- deoxyuridine) experiments were used to detect the effect of AEG-1 on the proliferation of PTC cells; Transwell test was used to detect the effect of AEG-1 on the metastasis of PTC cells; subcutaneous tumorigenesis test in nude mice was used to detect the effect of AEG-1 on the expression of autophagy related proteins and EMT related proteins. PTC cells in sh-NC group and sh-AEG-1 group were treated with rapamycin and transforming growth factor-β (TGF-β), respectively. CCK8 and transwell assay were used to detect the cell proliferation and metastasis ability of each group of cells, respectively. Results:Compared with normal thyroid cells Nthy-ori3-1, the expression level of AEG-1 in PTC cells was increased, with the highest expression in TPC-1 cells. After AEG-1 shRNA was transfected into TPC-1 cells, cell proliferation, metastasis and tumorigenicity in vivo were reduced ( P<0.05). Compared with the sh-NC group, the expression of autophagy-related proteins P62 and Beclin1 were increased and the expression of LC3B protein was decreased, and EMT-related proteins E-cadherin expression were increased and N-cadherin and Vimentin protein expression were decreased in the sh-AEG-1 group ( P<0.05). CCK8 and Transwell experiments showed that treatment with autophagy inducer Rapamycin and EMT inducer TGF-β attenuated the inhibitory effect of sh-AEG-1 on proliferation and metastasis ability of PTC cell ( P<0.05). Conclusions:AEG-1 promotes the proliferation and metastasis of PTC cells by inducing cell autophagy and EMT.

OBJECTIVE To establish a method for simultaneous determination of 15 bile acids in Tongren niuhuang qingxin pills， and to determine the contents of 15 batches of samples. METHODS Using dehydrocholic acid as internal standard， the determination was performed by ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method. The determination was performed on Hypersil GOLD C18 column with methanol-0.1% formic acid solution as the mobile phase by gradient elution at the flow rate of 0.2 mL/min. The column temperature was 40 ℃ ， and the sample size was 2 µL. Using heated electrospray ion source， parallel reaction monitoring mode scanning was performed in negative ion mode. SPSS 24.0 software was used for chemical pattern recognition analysis of content determination results. RESULTS The 15 bile acid components had a good linear relationship with peak area （all R2≥0.998 9）； their precision， repeatability and stability were all good （all RSD≤5.49%）； the average recoveries were 93.8%-105.7% （RSD was 0.5%-5.8%）. The average contents of taurocholic acid， 7-oxodeoxycholic acid， 12-dehydrocholic acid， glycocholic acid， 3-oxo-7α， 12α-hydroxy-5β-cholanoic acid， taurochenodeoxycholic acid， 3α-hydroxy- 7-oxo-5β -cholanic acid， hyocholic acid， taurodeoxycholic acid sodium salt hydrate， hyodeoxycholic acid， cholic acid， glycochenodeoxycholic acid， glycodeoxycholic acid， chenodeoxycholic acid and deoxycholic acid were 670.56， 25.97， 10.54， 280.12， 4.04， 29.81， 182.98， 813.55， 120.95， 220.31， 797.37， 18.37， 68.59， 30.13， 59.82 μg/g， respectively. Both cluster analysis and principal component analysis divided 15 batches of Tongren niuhuang qingxin pills into 2 categories， S1-S12 as one category and S13-S15 as the other category. CONCLUSIONS The established method is accurate， sensitive and specific， and can determine many types of bile acids. It also can quickly achieve the quantitative analysis of 15 bile acids in Tongren niuhuang qingxin pills， which is suitable for the quality control of this drug.