Objective To explore the protective effect of hesperidin pretreatment on concanavalin A (Con A)-induced acute liver injury and the effect on expression of TNF-α and IFN-γ. Methods Seventy-two SPF C57BL/ 6 mice were randomly divided into three groups: normal control group, model control group and hesperidin group. Acute liver injury model was established by injected with Con A. The hesperidin group was treated intragastrically with 1 000 mg·kg-1 hesperidin for 10 days. Model control group was treated intragastrically with the same volume of 0. 5% of sodium carboxymethyl cellulose. Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase ( AST) were measured. Pathological changes in hepatic tissue were observed under microscope. The expression of TNF-α and IFN-γ mRNAs in hepatic tissue was measured by reverse transcription polymerase chain reaction ( RT-PCR). The contents of TNF-α and IFN-γ in serum were detected by ELISA. Results Compared with model control group, the contents of ALT and AST in serum were significantly decreased (P<0. 01) in hesperidin group. Pathological changes in hepatic tissue were markedly improved. The expression of TNF-α and IFN-γ in the hepatic tissues and serum were significantly downregulated (P<0. 01). The concentrations of TNF-α and IFN-γ in hesperidin group were (717. 05±205. 22) and(611. 06±92. 82)pg·mL-1 in 2 h,(811. 56±167. 47)and(786. 19±215. 44)pg·mL-1 in 6 h. Compared with model control group, the expressions of TNF-α and IFN-γ in the hesperidin group were significantly downregulated (P<0. 01). But there was no significant difference between hesperidin group and model control group in 6 h after treated with Con A(P>0. 05). Conclusion Hesperidin pretreatment protects mice from Con A-induced acute liver injury possibly by inhibiting the expression of TNF-α and IFN-γ in the liver of mice.

Objective To investigate the effect of resveratrol (RSV) on epithelial-mesenchymal transition of MDA-MB-231 cells by down-regulating POLD1 expression. Methods CCK-8 was used to detect the effect of RSV on the activity of MDA-MB-231 cells. POLD1-OE and POLD1-NC cell lines were constructed by transfecting MDA-MB-231 cells with recombinant lentivirus. Western blot was used to detect the expression of POLD1, E-cadherin, N-cadherin and Vimentin after RSV treatment. Transwell invasion experiment and the scratch test were used to detect the cells invasion and migration abilities of each experimental group. Results RSV could significantly inhibit the survival of MDA-MB-231 cells, reduce the expression of POLD1, N-cadherin and Vimentin, increase the expression of E-cadherin, and inhibit the abilities of cell invasion and migration. Increasing the POLD1 expression could reduce the above-mentioned biological effects of RSV on MDA-MB-231 cells. Conclusion RSV could significantly inhibit the viability and EMT of MDA-MB-231 cells by down-regulating the expression of POLD1.

Objective To explore the effects and mechanism of triptolide on proliferation and apoptosis of breast cancer MCF-7 cells.Methods MCF-7 cells were treated by different concentrations of triptolide.CCK-8 as-say was employed to detect the cell proliferation. The morphological changes were observed by an inverted micro-scope.The apoptosis rate was detected by flow cytometry.Expressions of Bcl-2,Bax,Survivin and Caspase-3 were measured by qRT-PCR and Western blot. Results Triptolide inhibited the proliferation of MCF-7 cells in a dose and time-dependent manner at a suitable range.Triptolide induced morphological changes and apoptosis.Triptolide also down-regulated Bcl-2 and Survivin expressions and up-regulated Bax and Caspase-3 expressions. Conclu-sions Triptolide inhibits proliferation and induces apoptosis of MCF-7 cells,and its mechanism may be related to down-regulation of Bcl-2 and Survivin expressions and up-regulation of Bax and Caspase-3 expressions.

Objective: Primary and secondary school students in a unique stage of development, has a unique psychological age characteristics, their mental health screening should be appropriate for their psychological development characteristics. The contents of Middle School Students Mental Health Scale (MSSMHS) were explored and revised, and the reliability and validity of the revised scale were tested. Methods: The middle school and high school students in Shiyan city of Hubei province were selected by random cluster sampling, and the revised mental health scale for middle school students was used to investigate. The 57 items of the original scale were revised, and SPSS25.0 statistical software was used for statistical analysis of the data. Cronbach's Alpha coefficient under Alpha was used to test the reliability of the scale. Factor analysis was used for structural validity analysis. Results: The results of factor analysis, correlation analysis and internal consistency coefficient showed that the revised scale had high reliability, and its validity also met the requirements of psychological measurement. Conclusion The revised content of "MSSMHS" reasonably avoids cultural taboos and sensitive words of "suicide", and has good reliability and validity, which is worth popularizing in the practice of psychological assessment of middle school students.

Objective To explore the feasibility of 7,12?dimethylbenz[ a] anthracene ( DMBA) induced tree shrew breast cancer model, and compare the effects of two administration modes by gavage and mammary gland injection. Methods A total of 40 tree shrews were randomly divided into two groups (20 animals per group): DMBA gavage group and mammary gland injection group. DMBA was dissolved in edible vegetable oil. For gavage group, tree shrews were administered with DMBA solutions (15 mg／kg) by gavage once a day. For mammary gland injection group, DMBA solution (10 mg／kg) was injected into the mammary fat pad of tree shrews, and the injection was performed for a total of 3 times. From the first administration of DMBA, medroxyprogesterone acetate (MPA, 100 mg／kg) was intramuscularly injected into the muscles of the lateral thighs of tree shrews at the same time, for a total of 5 times. The tumorigenesis and survival of tree shrews were monitored. The tumor histological morphology was observed by HE staining. The expression of estrogen receptor (ER), progesterone receptor ( PR), cytokeratin5／6 ( CK5／6) and human epidermal factor receptor?2 (HER?2) was detected by immunohistochemical staining.Results In the gavage group, there were 10 deaths, and 4 tree shrews developed mammary tumors with 20.0%( 4／20) tumor formation rate. The success rate of mammary cancer modeling was 10.0%(2／20), and the tumor formation time was 197.3±15.1 days. In the mammary gland injection group, there were 8 tree shrews died, and 9 tree shrews formed tumors with 45.0%( 9／20) tumor formation rate. The success rate of mammary cancer modeling was 40.0%(8／20), and the tumor formation time was 71.8±19.0 days. There was no significant difference in mortality and tumor formation rate (P＞0.05) between the two groups (all P＞0.05). However, in the mammary gland injection group, the success rate of mammary cancer modeling was significantly higher than that in the gavage group (P＜0.05), whereas the tumor formation time was markedly shorter than that in the gavage group ( P＜0.01). The pathological types in the gavage group included ductal hyperplasia, intraductal papilloma and ductal carcinoma in situ, while those in the breast injection group included intraductal papilloma and ductal carcinoma in situ. In both groups, immunohistochemical staining showed the negative expression of HER?2 but positive expression of ER, PR and CK5／6 with varying degrees. Conclusion Both the DMBA gavage and mammary gland injection can successfully establish the tree shrew breast cancer model, and the modeling effect of mammary gland injection is better than gavage.

Objective:To investigate the effect of esculin on the proliferation of triple negative breast cancer cells and its molecular mechanism.Methods:MDA-MB-231 cells were treated with 28, 56, 112, 225, 450 and 900 μmol/L of esculin for 24, 48 and 72 h, respectively, and the cell viability was detected by cell counting kit 8 (CCK-8) assay. In addition, MDA-MB-231 cells were treated with 0, 225, 450 and 900 μmol/L of esculin for 48 h. And then the changes in cell morphology were observed by inverted microscope. The clone-forming ability was detected by colony formation assay. The mRNA expression levels of FBI-1, p53 and p21 were detected using real-time fluorescence quantitative polymerase chain reaction. The protein expression levels of FBI-1, p53, p21 and Ki67 were detected by western blot.Results:Compared with the blank control group, the cell viability of MDA-MB-231 cells that treated with esculin significantly decreased in a dose-dependent and time-dependent manners. After treatment with esculin, MDA-MB-231 cells shrunk, flattened, adhered poorly to the culture dish and the cell spacing became larger. Meanwhile, shedding and incomplete cells appeared, of which 900 μmol/L of esculin treatment group showed the most dramatic changes. In addition, the colony formation ratios were decreased to (77.18±5.13)%, (65.94±4.98)% and (45.92±3.70)% in the 225, 450 and 900 μmol/L of esculin treatment groups compared with blank control, respectively ( P<0.01). Furthermore, the mRNA and protein expressions of FBI-1 increased, while the levels of p53 and p21 mRNA and protein, as well as the protein expression of Ki67 decreased in a concentration-dependent manner ( P<0.01). Conclusion:Esculin may regulate cell cycle-related p53-p21 pathway via FBI-1 mediated DNA replication, thus inhibit the proliferation of triple negative breast cancer cells.

Objective To explore the feasibility of 7,12?dimethylbenz[ a] anthracene ( DMBA) induced tree shrew breast cancer model, and compare the effects of two administration modes by gavage and mammary gland injection. Methods A total of 40 tree shrews were randomly divided into two groups (20 animals per group): DMBA gavage group and mammary gland injection group. DMBA was dissolved in edible vegetable oil. For gavage group, tree shrews were administered with DMBA solutions (15 mg／kg) by gavage once a day. For mammary gland injection group, DMBA solution (10 mg／kg) was injected into the mammary fat pad of tree shrews, and the injection was performed for a total of 3 times. From the first administration of DMBA, medroxyprogesterone acetate (MPA, 100 mg／kg) was intramuscularly injected into the muscles of the lateral thighs of tree shrews at the same time, for a total of 5 times. The tumorigenesis and survival of tree shrews were monitored. The tumor histological morphology was observed by HE staining. The expression of estrogen receptor (ER), progesterone receptor ( PR), cytokeratin5／6 ( CK5／6) and human epidermal factor receptor?2 (HER?2) was detected by immunohistochemical staining.Results In the gavage group, there were 10 deaths, and 4 tree shrews developed mammary tumors with 20.0%( 4／20) tumor formation rate. The success rate of mammary cancer modeling was 10.0%(2／20), and the tumor formation time was 197.3±15.1 days. In the mammary gland injection group, there were 8 tree shrews died, and 9 tree shrews formed tumors with 45.0%( 9／20) tumor formation rate. The success rate of mammary cancer modeling was 40.0%(8／20), and the tumor formation time was 71.8±19.0 days. There was no significant difference in mortality and tumor formation rate (P＞0.05) between the two groups (all P＞0.05). However, in the mammary gland injection group, the success rate of mammary cancer modeling was significantly higher than that in the gavage group (P＜0.05), whereas the tumor formation time was markedly shorter than that in the gavage group ( P＜0.01). The pathological types in the gavage group included ductal hyperplasia, intraductal papilloma and ductal carcinoma in situ, while those in the breast injection group included intraductal papilloma and ductal carcinoma in situ. In both groups, immunohistochemical staining showed the negative expression of HER?2 but positive expression of ER, PR and CK5／6 with varying degrees. Conclusion Both the DMBA gavage and mammary gland injection can successfully establish the tree shrew breast cancer model, and the modeling effect of mammary gland injection is better than gavage.

Objective:To investigate the effect of esculin on the proliferation of triple negative breast cancer cells and its molecular mechanism.Methods:MDA-MB-231 cells were treated with 28, 56, 112, 225, 450 and 900 μmol/L of esculin for 24, 48 and 72 h, respectively, and the cell viability was detected by cell counting kit 8 (CCK-8) assay. In addition, MDA-MB-231 cells were treated with 0, 225, 450 and 900 μmol/L of esculin for 48 h. And then the changes in cell morphology were observed by inverted microscope. The clone-forming ability was detected by colony formation assay. The mRNA expression levels of FBI-1, p53 and p21 were detected using real-time fluorescence quantitative polymerase chain reaction. The protein expression levels of FBI-1, p53, p21 and Ki67 were detected by western blot.Results:Compared with the blank control group, the cell viability of MDA-MB-231 cells that treated with esculin significantly decreased in a dose-dependent and time-dependent manners. After treatment with esculin, MDA-MB-231 cells shrunk, flattened, adhered poorly to the culture dish and the cell spacing became larger. Meanwhile, shedding and incomplete cells appeared, of which 900 μmol/L of esculin treatment group showed the most dramatic changes. In addition, the colony formation ratios were decreased to (77.18±5.13)%, (65.94±4.98)% and (45.92±3.70)% in the 225, 450 and 900 μmol/L of esculin treatment groups compared with blank control, respectively ( P<0.01). Furthermore, the mRNA and protein expressions of FBI-1 increased, while the levels of p53 and p21 mRNA and protein, as well as the protein expression of Ki67 decreased in a concentration-dependent manner ( P<0.01). Conclusion:Esculin may regulate cell cycle-related p53-p21 pathway via FBI-1 mediated DNA replication, thus inhibit the proliferation of triple negative breast cancer cells.

Objective: To explore the feasibility of 7, 12-dimethylbenz[a] anthracene (DMBA) induced tree shrew breast cancer model, and compare the effects of two administration modes by gavage and mammary gland injection. Methods: A total of 40 tree shrews were randomly divided into two groups (20 animals per group): DMBA gavage group and mammary gland injection group. DMBA was dissolved in edible vegetable oil. For gavage group, tree shrews were administered with DMBA solutions (15 mg/kg) by gavage once a day. For mammary gland injection group, DMBA solution (10 mg/kg) was injected into the mammary fat pad of tree shrews, and the injection was performed for a total of 3 times. From the first administration of DMBA, medroxyprogesterone acetate (MPA, 100 mg/kg) was intramuscularly injected into the muscles of the lateral thighs of tree shrews at the same time, for a total of 5 times. The tumorigenesis and survival of tree shrews were monitored. The tumor histological morphology was observed by HE staining. The expression of estrogen receptor (ER), progesterone receptor (PR), cytokeratin5/6 (CK5/6) and human epidermal factor receptor-2 (HER-2) was detected by immunohistochemical staining. Results: In the gavage group, there were 10 deaths, and 4 tree shrews developed mammary tumors with 20.0% (4/20) tumor formation rate. The success rate of mammary cancer modeling was 10.0% (2/20), and the tumor formation time was 197.3±15.1 days. In the mammary gland injection group, there were 8 tree shrews died, and 9 tree shrews formed tumors with 45.0% (9/20) tumor formation rate. The success rate of mammary cancer modeling was 40.0% (8/20), and the tumor formation time was 71.8±19.0 days. There was no significant difference in mortality and tumor formation rate (P>0.05) between the two groups (all P>0.05). However, in the mammary gland injection group, the success rate of mammary cancer modeling was significantly higher than that in the gavage group (P<0.05), whereas the tumor formation time was markedly shorter than that in the gavage group (P<0.01). The pathological types in the gavage group included ductal hyperplasia, intraductal papilloma and ductal carcinoma in situ, while those in the breast injection group included intraductal papilloma and ductal carcinoma in situ. In both groups, immunohistochemical staining showed the negative expression of HER-2 but positive expression of ER, PR and CK5/6 with varying degrees. Conclusion Both the DMBA gavage and mammary gland injection can successfully establish the tree shrew breast cancer model, and the modeling effect of mammary gland injection is better than gavage.