[Effects of calponin-1 gene silencing on the biological behavior of uterine smooth muscle cells].

OBJECTIVE: To study the effects of calponin-1 expression inhibition on the proliferation , invasiveness, apoptosis and cytoskeleton of uterine smooth muscle cells, and explore the molecular mechanism of calponin-1 in the uterine smooth muscle cells for labor onset. METHODS: siRNA-calponin-1 adenovirus plasmid was constructed and transfected into primarily cultured uterine smooth muscle cells. The proliferation, invasiveness and apoptosis of the cells were determined by MTT assay, matrigel invasion assays and flow cytometry, respectively. Rhodamine-Phalloidin was used for labeling filamentous actin (F-actin), and the morphology and the distribution of F-actin was observed under fluorescence microscopy and analyzed quantitatively. RESULTS: The motor ability of uterine smooth muscle cells decreased significantly after transfection with siRNA-calponin-1 adenovirus plasmid (P<0.05). The transfected cells showed thinner, loosened and irregular F-actin microfibers, and the cells in the empty vector and blank control groups showed thicker and longer F-actin microfibers. CONCLUSION: Inhibition of calponin-1 expression can inhibit uterine smooth muscle cell migration and cause the morphological change and rearrangement of F-actin without affecting its proliferation and apoptosis in vitro, suggesting that the morphological change and rearrangement of F-actin of uterine smooth muscle cell may be one of the important mechanisms in the labor onset.

[Study of the role of nuclear factor-kappa B in preterm birth with subclinical chorioamnionitis].

OBJECTIVE: To investigate the role of nuclear factor-kappa B (NF-kappaB) in preterm birth with subclinical chorioamnionitis. METHODS: From October 2005 to October 2006, 111 cases including 36 cases of preterm birth in labor, 37 cases of full term gravida with spontaneous labor and 38 cases of full term gravida without threatened labor in the Hunan Province People's Hospital, third Xiangya Hospital of Central South University and Changsha Maternal and Child Care Service Center were enrolled in the study. After delivery, by pathology results of fetal membrane they were divided into two groups: subclinical chorioamnionitis group (subclinical infectious group) and non-infectious group. Immunohistochemical staining and RT-PCR were used to observe the change of the p65 subunit of NF-kappaB family in maternal blood and fetal membrane in subclinical infectious group and non-infectious group. RESULTS: (1) The incidence of subclinical chorioamnionitis: there were 24 cases of subclinical chorioamnionitis in the 36 cases of preterm birth in labor (67%), 7 cases in the 37 cases of full term gravida with spontaneous labor group (19%) and 3 cases in the 38 cases of full term gravida without threatened labor group (8%). There was a significant difference among the three groups (P < 0.01). In the totally 111 cases, 34 cases were classified as subclinical infectious group and 77 cases as non-infectious group. (2) In fetal membrane, the median of the average staining intensity of NF-kappaB p65 protein was higher in the subclinical chorioamnionitis group (8.0) than those in non-infectious group (4.0). Similarly, the average staining intensity of NF-kappaB p65 mRNA was higher in the subclinical infectious group (47.5 +/- 17.2) than those in non-infectious group (31.3 +/- 13.6). There was a significant difference between the two groups (P < 0.01). (3) In maternal blood, the expression of NF-kappaB p65 protein and mRNA was higher in subclinical chorioamnionitis group(8.0 and 42.6)than those in non-infectious group(4.0 and 23.6).There was a significant difference between the two groups (P < 0.01). (4) The concentration of NF-kappaB p65 protein in fetal membrane was positively correlated with that of maternal blood (r = 0.581, P < 0.01) and the concentration of NF-kappaB p65 mRNA in fetal membrane was positively correlated with that of maternal blood (r = 0.571, P < 0.01). CONCLUSION: The expression of NF-kappaB in maternal blood and fetal membrane in preterm birth with subclinical chorioamnionitis is higher and the two are correlated with each other. NF-kappaB p65 could be an accurate biochemical marker in predicting subclinical chorioamnionitis in preterm birth. NF-kappaB p65 plays an important role in the pathogenesis of subclinical chorioamnionitis in preterm birth.

[Killing activity of cytotoxic T lymphocytes stimulated by dendritic cell vaccine loaded with autologous cervical cancer antigen].

BACKGROUND & OBJECTIVE: Dendritic cells (DCs), the strongest antigen-presenting cells (APCs), can present antigens to T lymphocytes in vivo and in vitro, and induce cytotoxic T lymphocyte (CTL) reaction. This study was designed to investigate the killing activity of CTLs stimulated by Dcs loaded with autologous cervical cancer antigen in vitro. METHODS: Tumor antigens were made from frozen-thawed cervical cancer cells from patients after operation. DCs were isolated from peripheral blood mononuclear cells of patients with cervical cancer, cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4), loaded with tumor antigen to prepare DC vaccine, and used to stimulate autologous T lymphocytes to prepare antigen-specific CTLs. The killing activities of CTLs on autologous cervical cancer cells and HeLa, HepG2, MCF7, A549, and MGC803 cells were observed. RESULTS: CTLs stimulated by the DC vaccine had high killing activity on autologous cervical cancer cells, with killing rates of 79.32%-89.27% which were obviously higher than that of lymphokine-activated killing cells (t> or =2.89, P<0.05). The killing activity of CTLs was significantly weaker on HeLa cells (40.35%-58.09%) than on autologous cervical cancer cells (t> or =2.97, P<0.05). The specific CTLs had no obvious killing activity on HepG2, MCF7, A549, and MGC803 cells. CONCLUSIONS: CTLs stimulated by autologous cervical cancer antigen-loaded DCs have highly efficient and specific immune activity on autologous cervical cancer cells. It may be used in biotherapy for cervical cancer.