[Expression of vasohibin-1, MACC1 and KA11 proteins in serous ovarian cancer and their clinical significance].

OBJECTIVE: To examine the expression of vasohibin-1, metastasis-associated in colon cancer-1 (MACC1) and KAI1 proteins in serous ovarian cancer and their clinical significance.
 Methods: In 124 specimens of serous ovarian cancer (serous ovarian cancer group) and 30 specimens of ovarian serous cystadenoma (ovarian serous cystadenoma group), the expression of vasohibin-1, MACC1 and KAI1 protiens were detected by immunohistochemistry ElivisionTM method.
 Results: In the serous ovarian cancer group, the positive rates of vasohibin-1 and MACC1 proteins were 48.4% and 58.1%, respectively, which were both higher than those in the ovarian serous cystadenoma group (10.0% and 13.3%, respectively); while the positive rate of KAI1 protein in the serous ovarian cancer group was 33.9%, which was lower than that in the ovarian serous cystadenoma group (86.7%), there were significant differences between the 2 groups (all P<0.05). In the serous ovarian cancer group, the expression of the 3 proteins were closely related to the pathological grade, Federation International of Gynecology and Obstetrics (FIGO) stage and pelvic lymph node metastasis (all P<0.05). The KAI1 protein was negatively correlated with the levels of vasohibin-1 and MACC1 (r=-0.500, -0.600, respectively, both P<0.01); while there was a positive correlation between the vasohibin-1 and the MACC1 (r=0.518, P<0.01). Kaplan-Meier survival analysis showed that the over-expression of vasohibin-1, MACC1 and the low-expression of KAI1 protein were related to the survival rates (all P<0.05). Multi-factor analysis showed that the expression of vasohibin-1, KAI1 protein and the FIGO stage were independent prognosis factors for radical operation of serous ovarian cancer (RR=2.185, 3.893, 0.413; 95% CI=1.263-3.779, 2.190-6.921, 0.251-0.681; all P<0.05).
 Conclusion: The up-regulation of vasohibin-1, MACC1 and down-regulation of KAI1 in serous ovarian cancer are related to the tumor differentiation, clinical stage, metastasis and prognosis. Combined detection of these indexes is useful in predicting the progression and prognosis of serous ovarian cancer.

Porcine reproductive and respiratory syndrome virus inhibits MARC-145 proliferation via inducing apoptosis and G2/M arrest by activation of Chk/Cdc25C and p53/p21 pathway.

Porcine reproductive and respiratory syndrome virus(PRRSV) is an important immunosuppressive virus which can suppresses infected cells proliferation. In this work, we examined PRRSV ability to manipulate cell cycle progression of MARC-145 cells and explored the potential molecular mechanisms. The results showed that PRRSV infection imposed a growth-inhibitory effect on MARC-145 cells by inducing cell cycle arrest at G2/M phase. This arrest was due to the significant decrease of Cdc2-cyclinB1 complex activity in PRRSV-infected cells and the activity reduction was a result of Cdc2 Tyr15 phosphorylation and the accumulation of Cdc2 and cyclinB1 in the nucleus. Not only elevated Wee1 and Myt1 expression and inactivated Cdc25C, but also increase of p21 and 14-3-3σ in a p53-dependent manner caused the inhibitory Tyr15 phosphorylation of Cdc2. PRRSV infection also activated Chk1. Our data suggest PRRSV infection induces G2/M arrest via various molecular regulatory mechanisms. These results provide a new insights for PRRSV pathogenesis.

A Double-Antibody Sandwich ELISA for Sensitive and Specific Detection of Swine Fibrinogen-Like Protein 1.

Fibrinogen-like protein 1 (FGL1), a member of the fibrinogen family, is a specific hepatocyte mitogen. Recently, it has been reported that FGL1 is the main inhibitory ligand of lymphocyte activating gene 3 (LAG3). Furthermore, the FGL1-LAG3 pathway has a synergistic effect with programmed death 1 (PD-1)/programmed death ligand 1 (PD-L1) pathway and is regarded as a promising immunotherapeutic target. However, swine FGL1 (sFGL1) has not been characterized and its detection method is lacking. In the study, the sFGL1 gene was amplified from the liver tissue of swine and then inserted into a prokaryotic expression vector, pQE-30. The recombinant plasmid pQE30-sFGL1 was transformed into JM109 competent cells. The recombinant sFGL1 was induced expression by isopropyl-ß-d-thiogalactoside (IPTG) and the purified sFGL1 was used as an antigen to produce mouse monoclonal antibody (mAb) and rabbit polyclonal antibody (pAb). After identification, a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for sensitive and specific detection of sFGL1 was developed. Swine FGL1 in samples was captured by anti-sFGL1 mAb followed by detection with anti-sFGL1 rabbit pAb and HRP-conjugated goat anti-rabbit IgG. The limit of detection of the developed sFLG1-DAS-ELISA is 35 pg/ml with recombinant sFLG1. Besides, it does not show cross-reactivity with the control protein. Then serum samples of PRRSV-negative and -positive pigs were tested with the established DAS-ELISA and calculated according to the equation of y=0.0735x+0.0737. The results showed that PRRSV infection enhanced the serum FGL1 levels significantly. Our research provides a platform for the research on the functional roles of swine FGL1.

[Expressions of OCT4, Notch1 and DLL4 and their clinical implications in epithelial ovarian cancer].

OBJECTIVE: To investigate the correlations among OCT4, Notch1 and DLL4 and their association with the clinicopathological features of patients with epithelial ovarian cancer (EOC). METHODS: A total of 207 specimens of EOC and 65 specimens of benign ovarian epithelial tumor tissues were examined for expressions of OCT4, Notch1 and DLL4 proteins using immunohistochemistry. RESULTS: The positivity rates of OCT4, Notch1 and DLL4 in EOC tissues were 60.0%, 61.8% and 60.9%, respectively, significantly higher than the rates in benign epithelial tumor tissues (9.2%, 6.2%, and 0, respectively; P<0.05). The expressions of OCT4, Notch1 and DLL4 in EOC were significantly correlated with tumor differentiation, FIGO stage, and lymph node metastasis (P<0.05). DLL4 was positively correlated with OCT4 and Notch1 expressions (r=0.758 and 0.704, respectively, P<0.001), and the latter two were also positively correlated (r=0.645, P<0.001). Overexpressions of OCT4, Notch1 and DLL4 were associated with a poor prognosis, and the survival rate was significantly lower in patients positive for OCT4, Notch1, and DLL4 than in the negative patients (P<0.05). FIGO stage and expressions of OCT4 and DLL4 were independent prognostic factors of EOC (P<0.05). CONCLUSION: The expressions of OCT4, Notch1 and DLL4 are correlated with the differentiation, lymph node metastasis, clinical stage and prognosis of EOC. Combined detection of the 3 proteins has an important value in predicting the progression and prognosis of EOC.