Objective To explore the effect and potential mechanism of total lignans of syringae ramuls(TLS)against liver cancer u-sing in vitro cellular assays,network pharmacological and molecular docking.Methods Flow cytometry was performed to detect the effect of TLS on apoptosis and cycle of HepG2 cells.TL components were collected by literature search;drug-like properties and synthetic scores of components were assessed by ADMETlab;toxicological parameters of components were predicted by ProTox-Ⅱ;TLS component targets were predicted by Swiss Target Prediction website;GeneCards,OMIM,DisGeNET,and PharmGKB databases were used to screen the related targets of liver cancer;The"component-disease-target"network diagram was constructed by Cytoscape software and the"protein-protein interaction"(PPI)network diagram was constructed by STRING platform;and DAVID database was used to analyse GO,KEGG and WIKI metabolic pathways.Results TLS at 50-200μg/ml could significantly inhibit the proliferation of HepG2 cells,induce apoptosis and block the cell cycle.The main components of TLS in anti-liver cancer were 31 active components,such as(-)matairesinol,(-)-secoisolariciresinol,and pinnatifolin A,which mainly acted on 82 key targets such as Akt1,Bel-2 and EGFR,which were mainly enriched in the cancer pathway,EGFR tyrosine kinase inhibitor resistance,cycle and apoptosis signaling pathways.Conclusion TLS may play an anti-liver cancer effect through multi-components,multi-targets,and multi-pathway.

Aim To explore the potential mechanism of metformin on ATP-binding cassette transport A1(ABCA1)expression.Methods J774A.1 macrophages were treated with metformin and cycloheximide,and ABCA1 expression was determined by Western blot.His-tagged ABCA1 and HA-tagged Ub plasmids were co-transferred into HEK293 cells and stimulated with metformin.Co-immunoprecipitation(Co-IP)was used to test the binding ability of ABCA1 and ubiquitin.Candidate E3 ubiquitin-protein ligases(CE3)of ABCA1 were identified through Co-IP-based pro-teomics.The MIB1 plasmid was constructed and transferred into HEK293 cells,and Western blot was used to determine the effect of metformin and MIB1 on ABCA1 expression.Results Metformin increased the expression of ABCA1 in J774A.1 cells(P<0.01),and inhibited ABCA1 degradation(P<0.05).Metformin disrupted the binding of ABCA1 to ubiquitin(P<0.05).The proteins regulated by metformin in ABCA1 expression were primarily enriched in pathways re-lated to cell development,inflammation and immune defense.Metformin may upregulate ABCA1 protein expression via MIB1(P<0.05).Conclusion Metformin inhibits the degradation of ABCA1 by blocking the ubiquitin-proteasome system(UPS),and MIB1 might act as a candidate E3 ubiquitin-protein ligase(CE3)for ABCA1.

Objective To explore the effect and potential mechanism of total lignans of syringae ramuls(TLS)against liver cancer u-sing in vitro cellular assays,network pharmacological and molecular docking.Methods Flow cytometry was performed to detect the effect of TLS on apoptosis and cycle of HepG2 cells.TL components were collected by literature search;drug-like properties and synthetic scores of components were assessed by ADMETlab;toxicological parameters of components were predicted by ProTox-Ⅱ;TLS component targets were predicted by Swiss Target Prediction website;GeneCards,OMIM,DisGeNET,and PharmGKB databases were used to screen the related targets of liver cancer;The"component-disease-target"network diagram was constructed by Cytoscape software and the"protein-protein interaction"(PPI)network diagram was constructed by STRING platform;and DAVID database was used to analyse GO,KEGG and WIKI metabolic pathways.Results TLS at 50-200μg/ml could significantly inhibit the proliferation of HepG2 cells,induce apoptosis and block the cell cycle.The main components of TLS in anti-liver cancer were 31 active components,such as(-)matairesinol,(-)-secoisolariciresinol,and pinnatifolin A,which mainly acted on 82 key targets such as Akt1,Bel-2 and EGFR,which were mainly enriched in the cancer pathway,EGFR tyrosine kinase inhibitor resistance,cycle and apoptosis signaling pathways.Conclusion TLS may play an anti-liver cancer effect through multi-components,multi-targets,and multi-pathway.

OBJECTIVE To establish a content determination method for multiple components in Sophora flavescens from different origins and to evaluate its quality by combining with chemometrics. METHODS Thirteen batches （No. K1-K13） of S. flavescens from different origins were selected as test samples. A high-performance liquid chromatography-tandem triple quadrupole mass spectrometry （HPLC-MS/MS） method was established to determine the contents of 12 components， including matrine， oxymatrine， betaine， cytisine， N-methylcytisine， sophoridine， genistein， sophoricoside， sophorone， formononetin， sophorolone Ⅰ and norkurarinone in S. flavescens. Chromatographic separation was performed on a Shim-pack GIST-HP C18 column with a mobile phase consisting of methanol （A） and water containing 0.1% formic acid （B）， using gradient elution at a flow rate of 0.25 mL/min， column temperature of 35 ℃， and an injection volume of 3 μL. Mass spectrometry was conducted using an electrospray ionization source with positive and negative ion scanning. Data were collected in segments using the multiple reaction monitoring mode. Technique for order preference by similarity to ideal solution （TOPSIS） and grey relational analysis （GRA）methods were employed to compare and comprehensively evaluate the 13 batches of S. flavescens from different origins. RESULTS The methodological validation for the content determination met the relevant regulatory requirements. The contents of the 12 components were 490.66-1 231.00， 11 088.10- 18 021.50， 7.91-25.38， 903.97-1 713.64， 336.08-1 485.54，1 065.33-2 075.50， 27.52-71.80， 109.36-517.83， 6 034.55-10 632.73， 21.26-145.35， 814.84-1 911.32， 1 040.87-3 446.37 μg/g）， respectively. TOPSIS results showed that the top 7 samples in Euclidean distance ranking were K6， K12， K11， K3， K5， K10， K13. The GRA results showed that the top 7 samples in the relative correlation ranking were K12， K11， K10， K6， K13， K5， K3. CONCLUSIONS The established HPLC-MS/MS method is rapid， accurate， highly sensitive， stable and reliable. Combined with chemometrics methods， it can be used for the quality control and evaluation of S. flavescens. The comprehensive quality of samples K3， K5， K6（ from Hebei）， K10（ from Sichuan）， K11-K13（ from Shanxi）， etc. is relatively superior.

Aim To explore the potential mechanism of metformin on ATP-binding cassette transport A1(ABCA1)expression.Methods J774A.1 macrophages were treated with metformin and cycloheximide,and ABCA1 expression was determined by Western blot.His-tagged ABCA1 and HA-tagged Ub plasmids were co-transferred into HEK293 cells and stimulated with metformin.Co-immunoprecipitation(Co-IP)was used to test the binding ability of ABCA1 and ubiquitin.Candidate E3 ubiquitin-protein ligases(CE3)of ABCA1 were identified through Co-IP-based pro-teomics.The MIB1 plasmid was constructed and transferred into HEK293 cells,and Western blot was used to determine the effect of metformin and MIB1 on ABCA1 expression.Results Metformin increased the expression of ABCA1 in J774A.1 cells(P<0.01),and inhibited ABCA1 degradation(P<0.05).Metformin disrupted the binding of ABCA1 to ubiquitin(P<0.05).The proteins regulated by metformin in ABCA1 expression were primarily enriched in pathways re-lated to cell development,inflammation and immune defense.Metformin may upregulate ABCA1 protein expression via MIB1(P<0.05).Conclusion Metformin inhibits the degradation of ABCA1 by blocking the ubiquitin-proteasome system(UPS),and MIB1 might act as a candidate E3 ubiquitin-protein ligase(CE3)for ABCA1.

Objective To analyze the interactions between different structural types of volatile oil compo-nents(VOCs)and skin lipid molecules,and investigate the mechanism of volatile oil in Chi-nese materia medica(VOCMM)as penetration enhancers. Methods In this study,210 different structural types of VOCs were selected from the VOCMM penetration enhancer database,and the molecular docking experiments were conducted with three main lipid molecules of skin:ceramide 2(CER2),cholesterol(CHL),and free fatty acid(FFA).Each VOC was docked individually with each lipid molecule.Cluster analysis was used to explore the relationship between the binding energy of VOCs and their molecular struc-tures.Nine specific pathogen-free(SPF)Sprague Dawley(SD)rats were randomly divided in-to Control,Nootkatone,and 3-Butylidenephthalide groups for in vitro percutaneous experi-ments,with three rats in each group.The donor pool solutions were 3%gastrodin,3%gas-trodin+3%nootkatone,and 3%gastrodin+3%3-butylidenephthalide,respectively.The pen-etration enhancing effects of VOCs with higher binding energy were evaluated by comparing the 12-hour cumulative percutaneous absorption of gastrodin(Q12,μg/cm2). Results(i)Most of the VOCs were non-hydrogen bonded to the hydrophobic parts of CHL and FFA,and hydrogen bonded to the head group of CER2.Among them,sesquiterpene ox-ides showed the most pronounced binding affinity to CER2.The VOCs with 2-4 rings(in-cluding carbon rings,benzene rings,and heterocycles)demonstrated stronger binding affini-ty for three skin lipid molecules compared with the VOCs without intramolecular rings(P<0.01).(ii)According to the cluster analysis,most of the VOCs that bond well to CER2 had 2-3 intramolecular rings.The non-oxygenated VOCs were bonded to CER2 in a hydrophobic manner.The oxygenated VOCs were mostly bonded to CER2 by hydrogen bonding.(iii)The results of Franz diffusion cell experiment showed that the Q12 of Control group was 260.60±25.09 μg/cm2,and the transdermal absorption of gastrodin was significantly increased in Nootkatone group(Q12=5 503.00±1 080.00 μg/cm2,P<0.01).The transdermal absorption of gastrodin was also increased in 3-Butylidenephthalide group(Q12=495.40±56.98 μg/cm2,P>0.05).(iv)The type of oxygen-containing functional groups in VOCs was also an influencing factor of binding affinity to CER2. Conclusion The interactions between different types of VOCs with different structures in the VOCMM and three skin lipid molecules in the stratum corneum were investigated at the molecular level in this paper.This research provided theoretical guidance and data support for the screening of volatile oil-based penetration enhancers,and a simple and rapid method for studying the penetration-enhancing mechanism of volatile oils.

OBJECTIVE： To screen the hepatoprotective active fractions from Ixeris chinensis and study its chemical constituents. METHODS：The petroleum ether，ethyl acetate，n-butanol and residual water fractions from 70％ ethanol extract of I.chinensis were extracted by systematic solvent method. Human hepatocytes HL-7702 were induced by acetaminophen to induce liver injury model. MTT method was used to detect the protective effect of the above fractions（40 μg/mL，by the dosage of crude drug）on injured cells，and the active fractions were screened. The active fractions were separated and purified by silica gel column and Sephadex column chromatography. The structure of the compounds were identified by physical and chemical properties and spectral data （hydrogen spectrum，carbon spectrum）. RESULTS：After treated with different fractions of I. chinensis，the cell survival rate of each administration group was increased significantly，compared with model group（P＜0.01），and the n-butanol and water fractions had the strongest activity （the cell survival rates were 49.3％ and 52.2％ ，respectively）. Six compoundswere isolated from n-butanol fraction and identified as sonchifolignan A（Ⅰ），apigenin-7-O-β-D-glucopyranoside methyl ester（Ⅱ），luteolin-7-O-β-D-glucuronopyranoside methyl ester （Ⅲ），luteolin-7-O-β-D-glucopyranoside （Ⅳ），apigenin-7-O-β-Dglucopyranoside（Ⅴ）and luteolin（Ⅵ）. CONCLUSIONS：The n-butanol fraction is regarded as an effective position for protecting liver，and flavonoids are the main active omponents.KEYWORDS Ixeris chinensis；Hepatoprotective activi

OBJECTIVE：To optim ize ethanol extraction technology of Mongolian medicine Naru- 3. METHODS ：The L 9（34） orthogonal design was used to optimize ethanol extraction technology of Mongolian medicine Naru- 3 with solid-liquid ratio ，ethanol volume fraction and extraction time as factors ，using comprehensive scores for the contents of benzoylaconitine ，benzoylneoaconitine， benzoylhypoaconitine，aconitine，neoaconitine，hypoaconitine，piperine and gallic acid as indexes. RESULTS ：The optimal ethanol extraction technology was that solid-liquid ratio of 1∶10（g/mL），ethanol volume fraction of 75％，extracting for 1.5 h. After 3 times of validation tests ，average contents of above 8 components in ethanol extract from Naru- 3 were 1.69，1.48，14.69，0.28， 0.05，0.08，26.01，17.33 mg/g（RSDs were 0-4.96％，n＝3），respectively. Average comprehensive score was 19.03（RSD＝1.42％， n＝3）. CONCLUSIONS ：The optimal ethanol extraction technology of Mongolian medicine Naru- 3 is stable and feasible.

Introduction: In a report from the WHO 2013 it states that, 23% of children between the ages of 13 and 17 had suicidal ideation and 9.3% attempted suicide in the last 12 months. According to the research of Bayarmaa V et al, the prevalence of behavioral and emotional disorder among adolescents is between 8.7% & 9.4%. As a result of these researches, it can be concluded that evaluating the mental health of adolescents and learning the risk factors that can affect it has become an urgent matter in Mongolia. Goal: To establish the prevalence, the factors that influence it and the type of signs and symptoms common to this behavioral and emotional disorder in adolescents of the Gobi-Altai province. Material and Method: No ethical errors were reported during the implementation of this study. We used various versions of the Strengths and Difficulties Questionnaire (SDQ) applicable for children, adolescents, parents and teachers and determined exhibition of emotional and behavioral problems by cross-sectional analysis. 2192 adolescents between the ages of 11-18 years old, 1808 parents and caregivers and 102 teachers from the secondary school in Gobi-Altai province were screened from March to December of 2018. Results: In our study, 50.5% (n=1107) of participants were boys, 49.5% (n=1085) were girls and totally 2192 participants were involved. Regarding the survey results, 58.6% of adolescents in Gobi-Altai province were healthy, 36.1% of them had emotional and behavioral problems and 5.2% of them had emotional and behavioral disorders. For the mental health of adolescent, peer bullying (child 1.4 times higher, parents 6.4 times higher), moving house (child 2 times higher), domestic disputes between parents (child 1.6 times higher), loneliness (child 1.4 times high), hormonal change (teacher 7.7 times high, parents 2 times high), obtaining a qualification for a better life (teacher 6.4 times high) are the factors affecting their mental health negativ ely and creating a mental health problem. Conclusion 58.6% of adolescents in Gobi-Altai province were healthy, 36.1% of them had emotional and behavioral problems and 5.2% of them were with emotional and behavioral disorders. Peer bullying, moving house, domestic disputes between parents, loneliness, hormonal change are the factors affecting the mental health of adolescents negatively.