Objective To investigate the sensitization effect of Ailanthone (AIL) on cisplatin-resistant A549/DDP cells of non-small cell lung cancer. Methods The effects of AIL on the viability of A549 and A549/DDP cells were detected by MTT. Analysis of the combined drug use index(CI) of Ailanone and cisplatin using Chou-Talalay. Cell cycle and apoptosis were detected by flow cytometry, and the expression of apoptosis-related proteins were detected by Western blotting. Results AIL (0.6μmol/L) and DDP (50μg/mL) had synergistic effect. Cell cycle arrest and apoptosis rate increased in G1 phase. The protein expression of caspase-3 and bcl-2 decreased, but the protein expression of Cleaved Caspase3, Bax, CDK4 and CyclinD1 increased. Conclusion AIL enhanced the sensitivity of DDP to A549/DDP, and increased the growth inhibition and apoptosis of DDP on A549/DDP.

Objective To analyze the expression of miR-125b-5p in lung adenocarcinoma (LUAD), and predict its target genes and prognostic value by bioinformatics analysis. Methods Tissue and cell chips of cisplatin resistance-related LUAD in Gene Expression Omnibus (GEO) database were obtained, and differentially expressed miRNAs were screened by R software. The miRBase database was used to analyze conservativeness, and the dbDEMC3.0 database was used to evaluate the expression level. The target genes were predicted by TargetScan, miRDB and miRTarBase databases, and were analyzed for functional term enrichment with the DAVID database. Combined with the survival information of LUAD patients in the Cancer Genome Atlas (TCGA) database, prognostic analysis was performed. Quantitative Real-time PCR (qRT-PCR) detection technology was used to detect the expression level of miR-125b-5p in human normal bronchial epithelial cell line 16HBE, LUAD cell line A549, and cisplatin-resistance cell line A549/DDP. Results miR-125b-5p, closely related to cisplatin resistant of LUAD, was obtained by analysis of the chip data. The sequence of miR-125b-5p was highly conserved among species, and miR-125b-5p was significantly down-regulated in various tumors including LUAD. Fifty-four target genes of miR-125b-5p were predicted. The results of enrichment analysis showed that the target genes of miR-125b-5p were mainly involved in the regulation of gene expression, cellular macromolecule biosynthetic process, and mainly enriched on MicroRNAs in cancer, Protein processing in endoplasmic reticulum, and HIF-1 signaling pathway. Survival analysis indicated that miR-125b-5p was significantly associated with poor prognosis of patients with LUAD. qRT-PCR results showed that the expression level of miR-125b-5p in A549 was significantly lower than in 16HBE (P=0.008). Compared to parental cell, the expression level of miR-125b-5p was significantly down-regulated in A549/DDP cell (P=0.023). Conclusion miR-125b-5p was abnormally expressed in LUAD, and its target genes involved in multiple biological processes and signal pathways, which might be used as a new biomarker for prognosis in LUAD.

Objective To elucidate the long-term effect of isoproterenol on inducing anxiety-like behavior under chronic stress in kunming mice. Methods A total of 50 Kunming mice were randomly divided into control, chronic stress, isoproterenol (ISO) group, chronic stress +ICI 118, 551 (ICI) group and ISO+ICI 118, 551(ISO+ICI) group (n = 10 in each group). Control group: normal saline was intraperitoneally injected every day; Chronic stress group: Chronic stress treatment was given every day. ISO treatment group: ISO (5 mg/kg) was injected intraperitoneally every day. Chronic stress +ICI treatment group: ICI (0.2 mg/kg) was injected intraperitoneally every day after chronic stress treatment. ISO+ICI treatment group: ISO(5mg/kg) and ICI(0.2 mg/kg) were given intraperitoneally every day. Results The open arm residence time, the number of entering open arm, the percentage of open arm residence time and the number of entering open arm in the chronic stress group and ISO group were significantly lower than those in the control group (P<0.05). The open-box retention time, percentage of open-box retention time, open-box activity times and shuttle times in the chronic stress group and ISO treatment group were significantly lower than those in the control group (P<0.05). The retention time of dark box was significantly higher than that of the control group (P<0.05). Conclusion After long-term treatment with β-AR agonist isoproterenol, kunming mice developed anxiety-like behavior under chronic stress.

Objective To screen key genes and pathways associated with tumorigenesis and progression of triple-negative breast cancer (TNBC) by using bioinformatics analysis. Methods Two gene expression profilings containing TNBC (GSE76124) and normal mammary (GSE112825) tissue samples were obtained from Gene Expression Omnibus (GEO). R software was used to identify differentially expressed genes. Gene Ontology (GO) function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were further performed by DAVID. STRING and Cytoscape were used to construct protein-protein interaction(PPI) network. Furthermore, hub genes, core modules and seed genes were screened out. Results A total of 1,091 differentially expressed genes were screened out. The results of GO function analysis and KEGG pathway maps showed that these genes were associated with aromatic compound biosynthetic process, heterocycle biosynthetic process, and mainly enriched on the pathways in cancer and PI3K-Akt signaling pathway.We identified 10 hub genes, 2 core modules and 19 seed genes in PPI network. Conclusion A total of 10 hub genes and 19 seed genes were screened, which may play potential roles in the tumorigenesis and progression of TNBC. This study provids a bioinformatic analysis reference for further research.

Objective: To establish human choriocarcinoma JAR cells xenograft model in nude mice and observe its biological characteristics. Methods: 12 female Balb/c nude mice were randomly divided into normal control group and model group. The xenograft model was established by subcutaneous injection of choriocarcinoma JAR cells suspension; While the mice in normal control group were injected saline subcuatneouslly at the same site. The tumor formation rate and the survival time of nude mice were recorded; HE staining was used to observe the pathological changes of transplanted tumors and main organs, SP immunohistochemical staining to detect the β-human chorionic gonadotropin (β-HCG) expression in transplanted tumors of mice in model group, ELISA to detect the serum β-HCG level of mice in model group and normal control group. Results: The xenograft tumor formation rate in this study was 100% and the survival time of nude mice was (24.75±3.41) days. Gross morphological observation and histopathological observation showed that numerous trophoblastic proliferation with hemorrhagic necrosis in xenograft tumor, the cells markedly heteromorphic, with large and deeply stained nuclei. No obvious metastasis was observed in major organ. β-HCG protein positive expression could be observed in transplanted tumors of mice in model group. The serum β-HCG level of mice in model group was obviously higher than normal control group (P<0.05). Conclusions: The nude mice xenograft model is established using human choriocarcinoma JAR cells successfully in this study, which could provide an experimental animal model for choriocarcinoma studying.

Objective: To investigate the effects of dihydromyricetin on matrix metalloproteinase 2 (MMP-2), phosphatidylinositol3-kinase (PI3K) and protein kinase B (Akt) mRNA expression in choriocarcinoma JAR cells. Methods: Real time fluorescence quantitative PCR was used to detect the MMP-2, PI3K and Akt mRNA expression in JAR cells after the JAR cells were treated with dihydromyricetin (60μg/ml) for 36h; as well as the correlations between MMP-2 mRNA and PI3K, Akt mRNA in JAR cells were analyzed. Results: After the JAR cells were treated with dihydromyricetin (60μg/ml) for 36h, the MMP-2, PI3K and Akt mRNA expression in JAR cells significantly decreased compared with control group (P<0.05). Moreover, the MMP-2 mRNA expression in JAR cells was positively correlated with PI3K and Akt mRNA expression (r=0.957, 0.953, P<0.05). Conclusions: Dihydromyricetin may influence the invasion and metastasis of JAR cells by down regulating the MMP-2 gene expression in JAR cells, the mechanisms maybe related to the effects of dihydromyricetin on PI3K/Akt signaling pathway.

Objective: To investigate the effects of dihydromyricetin (DMY) on MicroRNA373 expression in choriocarcinoma JAR cells. Methods: JAR cells in logarithmic growth phase were treated with different concentrations of DMY (0μg/ml, 60μg/ml, 80μg/ml, 100μg/ml) for 24 hours, then the MicroRNA373 mRNA expression in JAR cells were detected by real time fluorescence quantitative PCR. Results: The MicroRNA373 expression of JAR cells in 3 DMY groups was all obviously higher than blank control group. Moreover, the MicroRNA373 expression gradually increased with increasing of DMY concentration (P<0.05). Conclusions: DMY can increase the MicroRNA373 expression in choriocarcinoma JAR cells in concentration dependent manner.