Objective:To study the effects of progesterone on the morphology, proliferation, and secretion of cytokines of human decidual stromal cells (DSCs) in late pregnancy, and to explore the mechanism of progesterone in preventing spontaneous preterm birth.Methods:Human decidual stromal cells in late pregnancy were cultured and treated with different concentrations of progesterone (in the experimental groups, 10 -6 mol/L, 10 -5 mol/L and 10 -4 mol/L progesterone was added to the culture medium respectively, and no progesterone was added to the culture medium of control group). The morphology of DSCs was observed under the microscope, the cell length/width ratio was measured, the proliferation was detected by methyl thiazolyl tetrazolium (MTT) method, and the expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected by RT-qPCR. Results:The length/width ratios of DSCs in progesterone 10 -4 mol/L (5.87±0.19) and 10 -5 mol/L (5.98±0.27) groups were lower than that in control group (6.42±0.19), the differences were statistically significant ( P<0.001, P=0.002). The length/width ratio in the 10 -4 mol/L group was lower than that in the 10 -6 mol/L group (6.28±0.32, P=0.005). The proliferation of DSCs in the 10 -5 mol/L (0.70±0.04) and 10 -4 mol/L (0.78±0.04) groups was higher than that in control group (0.59±0.05; P=0.027, P=0.002), and proliferation of DSCs in 10 -4 mol/L group was higher than that in 10 -6 mol/L group (0.61±0.01, P=0.004). The expression of TNF-α mRNA in each progesterone group was lower than that in control group (all P<0.001) and the expression of TNF-α mRNA in the 10 -5 mol/L and 10 -4 mol/L groups was lower than that in the 10 -6 mol/L group (all P<0.001) . The expressions of IL-6 mRNA decreased gradually in control group, 10 -6 mol/L, 10 -5 mol/L and 10 -4 mol/L groups, the differences were statistically significant (all P<0.001). Conclusion:Progesterone can make the decidual stromal cells wider, promote proliferation, and decrease the expressions of TNF-α and IL-6 mRNA, which may play an important role in the mechanism of progesterone preventing spontaneous preterm birth.

Objective:To study the effects of progesterone on the morphology, proliferation, and secretion of cytokines of human decidual stromal cells (DSCs) in late pregnancy, and to explore the mechanism of progesterone in preventing spontaneous preterm birth.Methods:Human decidual stromal cells in late pregnancy were cultured and treated with different concentrations of progesterone (in the experimental groups, 10 -6 mol/L, 10 -5 mol/L and 10 -4 mol/L progesterone was added to the culture medium respectively, and no progesterone was added to the culture medium of control group). The morphology of DSCs was observed under the microscope, the cell length/width ratio was measured, the proliferation was detected by methyl thiazolyl tetrazolium (MTT) method, and the expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected by RT-qPCR. Results:The length/width ratios of DSCs in progesterone 10 -4 mol/L (5.87±0.19) and 10 -5 mol/L (5.98±0.27) groups were lower than that in control group (6.42±0.19), the differences were statistically significant ( P<0.001, P=0.002). The length/width ratio in the 10 -4 mol/L group was lower than that in the 10 -6 mol/L group (6.28±0.32, P=0.005). The proliferation of DSCs in the 10 -5 mol/L (0.70±0.04) and 10 -4 mol/L (0.78±0.04) groups was higher than that in control group (0.59±0.05; P=0.027, P=0.002), and proliferation of DSCs in 10 -4 mol/L group was higher than that in 10 -6 mol/L group (0.61±0.01, P=0.004). The expression of TNF-α mRNA in each progesterone group was lower than that in control group (all P<0.001) and the expression of TNF-α mRNA in the 10 -5 mol/L and 10 -4 mol/L groups was lower than that in the 10 -6 mol/L group (all P<0.001) . The expressions of IL-6 mRNA decreased gradually in control group, 10 -6 mol/L, 10 -5 mol/L and 10 -4 mol/L groups, the differences were statistically significant (all P<0.001). Conclusion:Progesterone can make the decidual stromal cells wider, promote proliferation, and decrease the expressions of TNF-α and IL-6 mRNA, which may play an important role in the mechanism of progesterone preventing spontaneous preterm birth.

The aim of this study is to construct a bifunctional fusion protein, which can conjugate both human red blood cells and antibodies against classical swine fever virus (CSFV). We respectively amplified 2E8ScFv and mE2 genes from different recombinant vectors, in which 2E8ScFv gene is the single chain Fv gene against H antigen of human red blood cells, whereas mE2 gene is the main antigen coding region gene of CSFV E2 protein. We used overlap extension PCR to obtain an artificial fusion gene segment 2E8mE2 containing genes of Both 2E8ScFv and mE2, then ligated into the expression vector pET-DsbA and expressed in Escherichia coli BL21(DE3) PlysS host cells, after induced with IPTG the target fusion protein was successfully expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. We purified the fusion protein and renatured it from inclusion bodies to obtain a native state of well biological activity. The Erythrocyte agglutination test results indicated that the fusion protein can conjugate both human red blood cells and antibodies of CSFV.
Erythrocytes ; immunology ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunoglobulin Fragments ; biosynthesis ; genetics ; immunology ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Single-Chain Antibodies ; biosynthesis ; genetics ; Viral Envelope Proteins ; biosynthesis ; genetics