Effects of TLR9/NF-κB on oxidative stress and inflammation in IPEC-J2 cells.

BACKGROUND: Oxidative stress is one of the most important factors affecting large-scale breeding, especially the performance of pigs. Oxidative stress plays a role by affecting various genes in pigs, which can cause serious body damage, functional degradation and reduce production performance. OBJECTIVE: The purpose of this study was to investigate the effect of Toll like receptor 9 (TLR9) pathway on IPEC-J2 cells under oxidative stress and to provide reference for the growth development of Dapulian pigs. METHODS: In this study, Diquat was used as a source of oxidative stress to study the effects on Dapulian pigs by detecting relevant indicators. Then the IPEC-J2 cells were selected to verify the TLR9 signaling pathway in oxidative stress. RESULTS: Compared with the control group, superoxide dismutase (SOD) in experimental group decreased significantly, malondialdehyde (MDA) was significantly increased, accompanied by inflammatory reaction, and inflammatory factors were significantly increased in the experimental group. Oxidative stress model was constructed by H2O2 incubating IPEC-J2 cells. The interference and overexpression vectors of TLR9 and myeloid differentiation primary response protein 88 (MyD88) were constructed to detect the activity of antioxidant enzymes and related proteins. The results showed that overexpression of TLR9 enhanced the activity of antioxidant enzymes, decreased the secretion of inflammatory factors, and decreased the activity of MDA，reactive oxygen species (ROS); the results were opposite after TLR9 interference. This study also showed that H2O2 can activate the nuclear factor-κB (NF-κB) pathway and promote the translocation of NF-κB into the nucleus. After co-transfection with TLR9 and MyD88, the results showed that TLR9 regulated the expression of NF-κB through MyD88. CONCLUSION: The study showed that TLR9 pathway had a significant positive effect on antioxidant.

Two New 3D Supramolecular Complexes: Photocatalytic Properties and Treatment Activity for Gastric Cancer.

Two new coordination polymers, [Cu2(5-MeO-Hip)4(py)4]n (1) and [Co(5-EtO-ip)(4,4'-bipy)]n·n(MeOH) (2) (5-MeO-H2ip is 5-methoxyisophthalic acid, 5-EtO-H2ip is 5-ethoxyisophthalic acid, py is pyridine, and 4,4'-bipy is 4,4'-bipyridine), were created via solvothermal self-assembly. The thermal steadiness and photocatalytic functions of 1 and 2 were detected, and their application values in gastric cancer and the related mechanism were discussed. CCK-8 assay was used to determine the inhibitory activity toward gastric cancer cells' viability, and real-time RT-PCR was employed to examine the gastric cancer cells' Notch signaling pathway activity.

Role of IFNLR1 gene in PRRSV infection of PAM cells.

BACKGROUND: Interferon lambda receptor 1 (IFNLR1) is a type II cytokine receptor that clings to interleukins IL-28A, IL29B, and IL-29 referred to as type III IFNs (IFN-λs). IFN-λs act through the JAK-STAT signaling pathway to exert antiviral effects related to preventing and curing an infection. Although the immune function of IFN-λs in virus invasion has been described, the molecular mechanism of IFNLR1 in that process is unclear. OBJECTIVES: The purpose of this study was to elucidate the role of IFNLR1 in the pathogenesis and treatment of porcine reproductive and respiratory syndrome virus (PRRSV). METHODS: The effects of IFNLR1 on the proliferation of porcine alveolar macrophages (PAMs) during PRRSV infection were investigated using interference and overexpression methods. RESULTS: In this study, the expressions of the IFNLR1 gene in the liver, large intestine, small intestine, kidney, and lung tissues of Dapulian pigs were significantly higher than those in Landrace pigs. It was determined that porcine IFNLR1 overexpression suppresses PRRSV replication. The qRT-PCR results revealed that overexpression of IFNLR1 upregulated antiviral and IFN-stimulated genes. IFNLR1 overexpression inhibits the proliferation of PAMs and upregulation of p-STAT1. By contrast, knockdown of IFNLR1 expression promotes PAMs proliferation. The G0/G1 phase proportion in IFNLR1-overexpressing cells increased, and the opposite change was observed in IFNLR1-underexpressing cells. After inhibition of the JAK/STAT signaling pathway, the G2/M phase proportion in the IFNLR1-overexpressing cells showed a significant increasing trend. In conclusion, overexpression of IFNLR1 induces activation of the JAK/STAT pathway, thereby inhibiting the proliferation of PAMs infected with PRRSV. CONCLUSION: Expression of the IFNLR1 gene has an important regulatory role in PRRSV-infected PAMs, indicating it has potential as a molecular target in developing a new strategy for the treatment of PRRSV.

Effect of bevacizumab combined with docetaxel in the treatment of HER-2-negative recurrent metastatic breast cancer.

PURPOSE: To explore the efficacy and safety of bevacizumab combined with docetaxel in the treatment of human epidermal growth factor receptor-2 (HER-2)-negative recurrent metastatic breast cancer. METHODS: The clinical data of 128 patients with HER-2-negative recurrent metastatic breast cancer treated in our hospital from January 2015 to December 2016 were retrospectively analyzed. Sixty-four patients were treated with bevacizumab combined with docetaxel (Bevacizumab group), while the remaining 64 patients were treated with docetaxel alone (Docetaxel group). The clinical efficacy and adverse reactions were compared between the two groups, and the expressions of Ki-67, p53, matrix metalloproteinase-2 (MMP-2) and MMP-9 in breast cancer tissues were compared in both groups before and after treatment. The patient survival status and progression of disease were recorded through follow-up. RESULTS: In Bevacizumab group and Docetaxel group, the objective response rate (ORR) was 57.8% and 39.1%, and the clinical benefit rate (CBR) was 90.6% and 81.3%, respectively. The ORR was significantly better in Bevacizumab group than that in Docetaxel group. There was no statistically significant difference in the incidence rate of adverse reactions between the two groups. After treatment, the positive expression rates of Ki-67, p53, MMP-2 and MMP-9 obviously declined in both groups compared with those before treatment, showing statistically significant differences between the two groups. In Bevacizumab group and Docetaxel group, the mean overall survival (OS) was 13.3±5.5 months and 11.7±5.0 months, and the mean progression-free survival (PFS) was 7.1±2.6 months and 6.6±2.3 months, respectively. According to log-rank test, the OS rate was remarkably superior in Bevacizumab group to that in Docetaxel group (p=0.041), while the PFS rate had no statistically significant difference between the two groups (p=0.095). CONCLUSIONS: Bevacizumab combined with docetaxel has more excellent efficacy than docetaxel alone in the treatment of HER-2-negative recurrent metastatic breast cancer, and it prolongs the survival of patients, with tolerable adverse reactions, which is worthy of further clinical application.

[Construction of miR-331-3p overexpression vector and its effect on cell proliferation].

To investigate the effect of miR-331-3p on the proliferation of porcine renal epithelial cells (PK15) and its mechanism, the pcDNA 3.1(+) overexpression vector of miRNA-331-3p (pcDNA 3.1(+)-miR-331-3p) was constructed. PK15 cells were divided into four groups, including experimental group, experimental control group, inhibitor group and inhibitor control group. Experimental group and experimental control group were transfected with pcDNA 3.1(+)-miR-331-3p and pcDNA 3.1(+), respectively. Inhibitor group and inhibitor control group were transfected with miR-331-3p inhibitor and miR-331-3p negative control (miR-331-3p NC), respectively. Above all, CCK-8 reagent was used to plot the cell proliferation curve and Propidium (PI) staining was used to detect the proportion of cell stages. Secondly, its expression change were detected by quantitative real-time PCR that included the growth inhibitory protein family member 5 (ING5), cyclin-dependent kinase 2 (CDK2), cyclin-dependent kinase 3 (CDK3), cyclin-dependent kinase 4 (CDK4), Cyclin B and cyclin-dependent kinase inhibitor 1A (CDKN1A). The results showed that the expression of miRNA-331-3p was significantly increased in the experimental group. The cell proliferation curve showed that the number of cells in experimental group was significantly higher than that in experimental control group or inhibitor control group at 48 h and 72 h (P<0.05). Simultaneously, Inhibitor group was significantly lower than experimental control group or inhibitor control group in the number of cells at 48 h and 72 h (P<0.05), but there was no significant difference between the experimental group and the control group. Compared with the experimental control group, the proportion of cells of experimental group in G0/G1 phase decreased, the proportion of S phase and G2/M phase increased, and the inhibitor control group showed the opposite trend. Simultaneously, the expression levels of CDK2, CDK3, CDK4 and Cyclin B genes in the experimental group were significantly increased, while ING5 and CDKN1A genes inhibiting proliferation showed a significant downward trend. These results demonstrate that the miR-331-3p overexpression vector was successfully constructed, and miR-331-3p has the ability to promote the proliferation of PK15 cells. The study lays a solid foundation for further research for its role in pig growth and development.