Objective To investigate the protective effect of insulin-like growth factor-1 (IGF-1) on 6-hydroxy dopamine (6-OHDA) induced oxidative damage in PC-12 cells. Methods PC12 cells were treated with 6-OHDA (concentrations of 25, 50, 100, 150 and 200 μmol/L). In order to select the optimal experimental concentration and treatment time, the activity of PC12 cells was detected by MTT at different time points of 12 h, 24 h and 48 h after treatment. PC12 cells were divided into three groups: control group, 6-OHDA group and IGF-1+6-OHDA group. The activity of PC12 cells was detected by MTT assay. Reactive oxygen species (ROS) level of PC12 cells was detected by immunofluorescence staining, and apoptosis was detected by Hoechst33342 / PI double staining method. Results With the increased concentration and the prolongation of the action time of 6-OHDA, the activity of PC12 cells decreased gradually. The concentration of 150 μmol/L and action time of 24 h of 6-OHDA were selected as the optimal experimental concentration and observation time for this study. Compared with 6-OHDA group, the activity of PC12 cells increased, the expression level of ROS and the apoptosis decreased in IGF-1+6-OHDA group. Conclusion IGF-1 pretreatment can reduce 6-OHDA induced oxidative damage and apoptosis of PC12 cells, also can increase cell activity, which can provide a potential strategy for the prevention and treatment of Parkinson’s disease.

Objective To investigate the inhibitory effect and molecular mechanism of delphinidin (Dp) on HER-2 positive breast cancer. Methods The HER-2 positive breast cancer cells (MDA-MB-453 and BT-474) were treated with different concentrations of delphinidin (10，20，40，80 and 160 μmol/L) for 48 hours. The same concentration of DMSO was used as the solvent control. CCK-8 method was used to measure the effect of Dp on cell activity, and half maximal inhibitory concentration (IC50) was calculated to determine the concentration of subsequent experiments. The cells were treated with different concentrations of Dp (20, 40 and 80 μmol/L) for 48 hours. HE staining was used to observe the cell morphological changes. Flow cytometry was used to analyze the cell cycle. Cell apoptosis rate was detected by TUNEL assay. The phosphorylation levels of NF - κB signaling pathway related proteins were determined by Western blot assay. Results Delphinidin inhibited the proliferation of MDA-MB-453 and BT-474 in the concentration ranges of 10，20，40，80 and 160 μmol/L (IC50: 41.02 and 60.97 μmol/L). In the concentrations of 20，40 and 80 μmol/L, compared with the control group, it was found that some cells were detached, floated and lysed, and the cell volume was decreased, the proportion of cells in G2/ M phase and the apoptosis rate were increased in DP treatment groups. Compared with the control group, the expression levels of p-NF-κB/p65, p-IκBα, p-IKKα/β and p-PKCα were significantly decreased in the 40 and 80 μmol/L Dp treatment groups, while the expression levels of IκBα, IKKα, IKKβ and PKCα were increased in the Dp treatment groups (P＜0.05). Conclusion Delphinine can inhibit the proliferation of breast cancer cells by blocking the NF-κB signaling pathway and inducing the G2/M cycle arrest and apoptosis of MDA-MB-453 and BT-474 cells. Key words：receptor, epidermal growth factor; antineoplastic agents, phytogenic; breast neoplasms; cell cycle; delphinidin;HER-2 positive breast cancer; NF-κB signaling pathway

Objective To explore the potential pathogenic mechanism of adrenocortical carcinoma (ACT), and screen out genes that may be related to biological targets. Methods In this study, the gene expression datasets of ACT were obtained from the Gene Expression Omnibus (GEO) with the accession number of GSE75415. Through R programming software, the microarray preprocessing and differential expression analysis of 18 ACT tissue samples (experimental group) and 7 normal adrenocortical tissue samples (control group) were conducted to identify potential biomarkers for ACT in different stages. Besides, through functional enrichment and Kaplan-Meier analysis, several more reliable biomarkers for ACT were identified. At the same time, the two generation sequencing data of the TCGA database, including 79 ACT samples were analyzed, and the genes that can affect the survival of ACT patients were screened. Results There were 248, 334, 315 and 561 differentially expressed genes in stage1-4 respectively. There were 73 overlapping genes (OLDEGs) among the different grading samples. Central genes HSPA13, GARS, STXBP1, AKIRIN1 and TUBB3, were up-regulated in all of stages of ACT samples compared with those of normal samples, while, central genes ADH1B, DCN, RASSF2, PDGFRA, PLAT, C3 and FOS were down-regulated in ACT samples. They were found to be significantly associated with pathways of immune response, cell cycle, phosphorylation and cruor, which were all closely related with ACT progression. Besides, Kaplan-Meier analysis of 73 OLDEGs in 79 ACT samples from TCGA database identified several genes, including XPO1, RACGAP1, PDGFD, NR4A2, MXRA5, VPS51, TMED3, NDFIP1 and CDKN1C, which were significantly associated with ACT overall survival. Conclusion Differentially expressed genes and survival related genes in all of stages can serve as new targets for ACT therapy, and which should be helpful for the understanding of its pathogenesis and prognosis.

Objective To investigate the effect of dual-specificity phosphatase-2 (DUSP2) on the cell proliferation and apoptosis in the gastric cancer and its mechanisms. Methods Firstly, the effects of different expressions of DUSP2 on the overall survival of 876 gastric cancer patients were analyzed by online analysis tool KM plotter, and the expressions of DUSP2 in various gastric cancer cell lines (MKN-45, SGC-7901, HGC-27 and N-87) were verified. Secondly, DUSP2 overexpressed lentiviral vector was constructed, and MKN-45 was transfected by packaged virus. DUSP2-overexpression gastric cancer cell line was gained by drug screening. Meanwhile, gastric cancer cells infected with empty vector virus were used as control. Then the effect of DUSP2 upregulation on the proliferation ability of gastric cancer cells was evaluated by MTS cell proliferation assay, and the apoptosis was determined by Annexin V-FITC / PI double staining. The protein expressions of DUSP2, ERK, p-ERK (Thr202/Tyr204), P38 and p-P38 were tested by the Western blot analysis. Results Gastric cancer patients with high DUSP2 expression showed a significant survival advantage compared with those with low DUSP2 expression, and DUSP2 levels were decreased in several gastric cancer cell lines. The Western blot analysis revealed that the expression of DUSP2 markedly increased in overexpressed DUSP2 group (experimental group) compared with that of control group. The MTS experiment showed that the cell viability was significantly decreased in experimental group than that of the control group. Correspondingly, the cell apoptosis test showed that the cell apoptosis rate was obviously higher in the experimental group than that of the control group. The results of Western blot assay indicated that p-ERK (Thr202/Tyr204) and p-38 were significantly down-regulated in the experimental group compared with those of control group. Conclusion The over-expression of DUSP2 can efficiently inhibit cell proliferation and promote its apoptosis in gastric cancer cells, and the mechanism is related to DUSP2 inhibiting the phosphorylation levels of ERK and P38.

Objective To investigate the dynamics of cortical granules exocytosis (CGE), and the role of Synaptotagmin1 (Syt1) in mouse fertilization. Methods The dynamics of mouse CGE were filmed on Perkin Elmer precisely Ulta VIEW VOX confocal Imaging System. The Syt1 functions on mouse fertilization and expression in zygotes were analyzed by immunofluorescence, Western blot assy and qRT-PCR. Results The dynamic process of mouse CGE was observed after oocyte activation, and the exact pot seems to be close to the polar body. The Syt1 expression was gradually increased after fertilization. The mouse fertilization rate after Syt1 knockdown was significantly lower than that of the control group (P＜ 0.05). Conclusion The dynamic process of CGE is studied. It is found that Syt1 is involved in mouse fertilization process.

Objective To observe the effect of Xuebijing on immunosuppressed sepsis of model mice. Methods The 152 mice were randomly divided into control group (Control), immunosuppression group (IM), immunosuppressed sepsis model group (ISM) and Xuebijing treatment group (XT). There were thirty – eight mice for each group. For group IM, cyclosporine A was injected intraperitoneally along the median line of the lower abdomen for immunosuppression, 25 mg/kg， 1 time every other day for a total of 3 times. For group ISM, after immunosuppression, 300 μL escherichia coli 44102 with a concentration of 1×109 CFU/mL was injected intraperitoneally along the midline of the lower abdomen. The 30 minutes after the establishment of the immunosuppressive sepsis model, Xuebijing 4 mL/kg was intraperitoneally injected along the midline of the lower abdomen in the ISM group. After 30 minutes, the same dose of Xuebijing injection was repeated once. Control group was injected intraperitoneally with the same amount of normal saline. (1) After 8 h, 4 mice in each group were taken for blood bacterial culture. (2) After 12 h, 10 mice in each group were taken for detecting CD3+CD4+ and CD3+CD8+ in peripheral blood using flow cytometry. (3) After 12 h, 10 mice in each group were taken for detecting blood levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) by enzyme-linked immunosorbent assay (ELISA). (4) After 12 h, 4 mice in each group were taken for detecting high-mobility group protein (HMGB1) by Western blot assay. (5) After 12 h, 10 mice in each group were taken for detecting alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN) and serum creatinine (CR) levels by automatic biochemical analyzer. Results Compared with the ISM group, the ratio of CD3+CD4+/CD3+CD8+ was significantly increased, the number of blood bacteria culture decreased obviously in the XT group. Liver and kidney function indicators ALT, AST, CR, BUN and inflammatory factor indicators TNF-α, IL-6 and HMGB1 were also significantly decreased in the XT group. Conclusion Xuebijing can obviously modulate the immunosuppression, against bacteria, inhibit the inflammatory reaction, and protect the vital organs.

Objective To explore the relationship between expressions of high mobility group B1 and N1(HMGB1 and HMGN1) and clinical pathological parameters and prognosis in patients with non-small cell lung cancer (NSCLC). Methods Ninety-one postoperative tumor tissue specimens from the patients with NSCLC were collected in Tianjin Medical University Cancer Institute and Hospital from January 2004 to May 2011. Immune histochemical assay was used to detect the expressions of HMGB1 and HMGN1. According to the staining intensity, expressions of HMGB1 and HMGN1 were divided into positive and negative groups. Kaplan-Meier was used to analyze the correlation between the expressions of HMGB1/HMGN1 and clinical pathological parameters/prognosis. Results The cytoplasmic expressions of HMGB1 (40%, 36/91) was positively correlated with cytoplasmic expression of HMGN1 (31%, 28/91, rs=0.319，P<0.001). The expression level of HMGN1 was significantly higher in patients with late stage of NSCLC (Ⅲ~Ⅳ) than that in patients with early stage of NSCLC (Ⅰ~Ⅱ, P＜0.05). It was also found that the expression level of HMGN1 was significantly higher in patients with lymph node metastasis than that in patients without lymph node metastasis (P＜0.05). The poor prognosis of NSCLC was slightly lower in patients with high expression of HMGN1 than that in patients with low expression of HMGN1, but the difference was not statistically significant. Conclusion The HMGN1 can be used as a promising prognostic biomarker for predicting the prognosis of NSCLC patients.

Objective To investigate the relationship between single nucleotide polymorphisms (SNPs) of betaine homocysteine methyltransferase (BHMT) and reduced folate carrier 1 (RFC1, SLC19A1) genes and susceptibility to neural tube defects (NTDs) in children of Han population in northern China. Methods A total of 152 NTDs patients, and 169 controls were used in this study. Sequenom Mass-ARRAY was used to genotype 9 SNPs in 2 genes, and Haploview 4.2 software was used for Haplotype analysis. Results The allele frequency of rs3733890 in BHMT was significantly correlated with the incidence of NTDs, and the children with A allele were at higher risk of NTDs compared with those with G allele (OR=1.408, 95%CI: 1.013-1.956). The distribution of allele and genotype frequencies of rs1051266 in RFC1 was significantly associated with the incidence of NTDs, and the children with G allele and GG genotype were associated with higher risk of NTDs compared with those with A allele and AA genotype (OR=1.492, 95%CI: 1.089-2.044; OR=2.020, 95%CI: 1.081-3.780). The allele frequency of rs3788200 in RFC1 was significantly correlated with the incidence of NTDs, and the children with G allele were at higher risk of NTDs than those with A allele (OR=1.368, 95%CI: 1.002-1.868). Meanwhile, Haplotype analysis showed that C-A-A-A haplotype (rs567754-rs3733890-rs558133-rs585800) of BHMT and G-G-G-T haplotype (rs1051296-rs3788200-rs1051266-rs4819130) of RFC1 were associated with the increased risk of NTDs, but C-G-A-A haplotype of BHMT gene was associated with the decreased risk of NTDs. Conclusion The rs3733890 of BHMT gene, rs1051266 and rs3788200 of RFC1 gene are associated with susceptibility to NTDs in children of Han population in northern China.

Objective To compare the clinical outcomes of the first and the second-generation drug-eluting stents (DES) implanted in saphenous vein grafts (SVG) in patients after coronary artery bypass graft (CABG). Methods A total of 108 patients with coronary angiography and DES implanting in SVG due to ischemia symptoms after CABG were collected in this study, including 69 patients with the first-generation of DES (drug-eluting: sirolimus) and 39 patients with the secondgeneration of DES (drug-eluting: zotarolimus or everolimus). The success rate of stents and mortality in hospital were compared between two groups of patients. The major adverse cardiac events (MACE), such as all-cause death, target vessel revascularization (TVR) and acute myocardial infarction (AMI) in 2-year follow-up were also compared between the two groups of patients. The survival curve was drawn by Kaplan-Meier method, and the MACE free survival rates of two groups of patients were compared. Cox regression analysis was used to evaluate the risk factors for MACE in patients with SVG stent implantation. Results There were no significant differences in the success rate of stents and mortality in hospital between the two groups. In average 2-year follow-up, a total of 37 cases of MACE were performed. There was no statistical difference in the incidence of MACE between the two groups (34.8% vs. 33.3%, P＞0.05). The proportion of TVR was significantly lower in the second-generation group than that of the first-generation group (13.0% vs. 2.6%, P＜0.05). Kaplan-Meier survival analysis showed that there were no statistically differences in the survival rates of no-cumulative events between the two groups (81.2% vs. 79.5%, Log-rank χ2=0.029, P＞0.05). COX regression analysis showed that diabetes (HR=2.530, 95% CI: 1.008-6.345, P=0.041) and stent diameter (HR=1.143, 95% CI: 1.043-1.253, P=0.004) were independent predictors for the MACE in patients implanted stents in SVG. Conclusion There are no significant differences in mortality in hospital and the MACE in 2-year follow-up between the patients of two generations of DES implanting in the SVG after coronary artery bypass grafting. The proportion of TVR is lower in the second-generation DES group. Patients with diabetes and large diameter stents have a poor prognosis.

Objective To explore the pathogen distribution, changes of cardiorespiratory function and changes of serum inflammatory factors in old patients with cardiac failure complicated with pulmonary infection. Methods A total of 153 cardiac failure patients hospitalized in our hospital from March 2014 to September 2017 were retrospectively analyzed. The patients were divided into pulmonary infected group (n=76) and non-infected group (n=77). Another 82 healthy subjects were served as control group. The respiratory secretions were collected to detect pathogen distribution in infected group. The changes of cardiac and pulmonary functions and peripheral blood inflammatory factors were compared between the three groups. Results A total of 110 pathogens were isolated from infected group, which contained 81 (73.63%) gram negative strains, 27 (24.55%) gram positive strains and 2 (1.82%) fungus. Compared with the control group and non-infected group, the lung function index and pulmonary function decreased significantly in the infected group, containing forced expiratory volume in one second (FEV1), forced vital capacity (FVC), FEV1/FVC, carbon monoxide diffusing capacity (DLCO), maximum mid expiratory flow (MMEF), left ventricular end-diastolic dimension (LVEDD) and left ventricular end-systolic dimension (LVESD). Meanwhile, the serum levels of tumor necrosis factor (TNF) –α, interleukin (IL)-6 and procalcitonin (PCT) were significantly higher in the infected group than those in the non-infected group and the control group (P＜0.05). Conclusion Gram negative strains dominate in pulmonary infection of old patients with cardiac failure, and the cardiopulmonary function is significantly decreased due to the infection, whereas the serum levels of inflammatory factors are dramatically increased, which has an auxiliary value in the evaluation of pulmonary infection.