Association between DSCR4 gene methylation in plasma in early pregnancy and Down's syndrome.

Down's syndrome (DS), a chromosomal abnormal genetic disease caused by a local or total copy of chromosome 21, leads to patients suffering from delayed body growth, special facies, mild to moderate mental retardation and other symptoms, seriously affecting the life of patients. The aim of the present study was to examine the association between Down's syndrome critical region 4 (DSCR4) gene methylation in plasma in high-risk pregnant women with DS in early pregnancy (hereinafter referred to as pregnant women in early pregnancy) and DS, in order to screen new epigenetic markers for the clinical diagnosis of DS. DNA in peripheral blood cells and plasma in pregnant women in early pregnancy were treated with hydrosulphite. DSCR4 genes with different methylation levels were amplified by methylation-specific polymerase chain reaction (PCR), and the methylation difference of the CpG site of the DSCR4 amplification product in peripheral blood DNA was verified via restriction endonuclease analysis. The expression of DSCR4 with different methylation levels in peripheral blood of pregnant women in early pregnancy were detected via reverse transcriptase-quantitative PCR (RT-qPCR), and the DSCR4 gene functions were studied via the intervention in DSCR4 expression with small interfering RNA (siRNA). Methylation-specific PCR and restriction endonuclease analysis revealed that DSCR4 genes were differentially methylated in peripheral blood DNA in pregnant women in early pregnancy. Additionally, DSCR4 showed a low methylation status in plasma but a high methylation status in peripheral blood cells. RT-qPCR revealed that non-methylated DSCR4 was highly expressed in the peripheral blood of pregnant women in early pregnancy, and thus was an epigenetic marker of fetal DS. siRNA results showed that the downregulation of DSCR4 inhibited cell migration and invasion, but had no effect on cell proliferation. The results suggest that the DSCR4 gene was differentially methylated in peripheral blood DNA in pregnant women in early pregnancy. Furthermore, DSCR4 exists in a non-methylated state in plasma and in a hyper-methylated state in blood cells. DSCR4 can therefore promote the migration and invasion of trophocytes and serve as an epigenetic marker of non invasive clinical diagnosis of DS.

APOBEC3 deletion polymorphism is associated with epithelial ovarian cancer risk among Chinese women.

Ovarian cancer remains the leading cause of death from gynecological malignancies and the second most common gynecological malignancy among women worldwide. However, the etiology still remains largely unknown. Previous studies identified APOBEC3 gene deletions were significantly associated with higher breast cancer risk in both European and Chinese women. Considering both breast and ovarian cancers being hormonally driven and sharing multiple risk factors, we performed this case-control study to evaluate the association between APOBEC3 deletion and epithelial ovarian cancer (EOC) risk. We analyzed the APOBEC3 deletion in a case-control study of 2,938 Chinese women (including 1,374 EOC cases and 1,564 healthy controls). All participants were genotyped using real-time qualitative PCR (qPCR). APOBEC3 deletion was significant associated with EOC risk, with ORs (95 % CIs) of 1.46 (1.14-1.86) associated with one copy deletion and 2.53 (0.91-7.06) associated with two copy deletion compared with subjects with no deletion (P for trend =1.50 × 10(-3)). Additional adjustments and stratified analyses did not change the results materially. Our data suggests that the loss genotypes of APOBEC3 deletion predispose their carriers to EOC.

[Identification of contraction related proteins in corpus myometrium at labor].

OBJECTIVE: To explore the molecular mechanism of human corpus myometrium contraction related proteins and parturition. METHODS: The proteins of human corpus of myometrium tissues from full term non-in labor (38- 41 weeks amenorrhea) and full term in labor (38-41 weeks amenorrhea) gravidas were separated by 2-dimensional gel electrophoresis (2-GE), respectively. Then gels were stained by Coomassie brilliant blue G250, scanned by Image scanner and analyzed with PDQuest software. The differentially expressed protein spots of corpus myometrium between the 2 groups were identified by peptide mass finger print based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. Three identified differentially expressed proteins were confirmed by Western blot. RESULTS: Well resolved and reproducible 2D-GE maps of human corpus myometrium from non-in labor and in-labor gravidas were acquired. Twenty more than 2-fold differentially expressed protein spots were identified by MALDI-TOF-MS. These proteins were involved in cell structure, calcium binding, chaperone, energy metabolism, signal transduction, and antioxidant. CONCLUSION: Twenty contraction related proteins of human corpus myometrium have been identified, indicating that cell structure and calcium-binding are important reasons for the contraction of human corpus myometrium.

The involvement of osteopontin and ß3 integrin in implantation and endometrial receptivity in an early mouse pregnancy model.

OBJECTIVE: To explore the roles of osteopontin and ß3 integrin in successful implantation. STUDY DESIGN: In this study, an early pregnant mouse model was established by peritoneal injection of pregnant mare serum gonadotropin and human chorionic gonadotropin (PMSG+hCG). The expression of osteopontin (OPN) and ß3 integrin on the endometrium was measured by immunohistochemistry, RT-PCR, and western blot. The function of OPN and ß3 integrin in implantation was investigated by intrauterine injection of OPN and ß3 integrin antibody. RESULTS: We found that PMSG+hCG injection significantly increased the number of blastocysts during implantation as well as the concentration of estradiol and progesterone in serum and endometrium tissues. OPN and ß3 integrin were co-expressed in luminal epithelium and their levels dynamically changed from day 4 to day 8 of pregnancy with peak expression on day 5. The percentages of OPN and ß3 integrin positive cells in the luminal epithelium were significantly higher in PMSG+hCG-stimulated mice on day 5 than in control mice. Functional blockade of OPN and ß3 integrin significantly inhibited implantation. CONCLUSIONS: This study suggests that co-expression of OPN and ß3 integrin is a biological marker for good endometrial receptivity and that both proteins play a crucial role in blastocyst implantation.

Reversal of paclitaxel resistance in epithelial ovarian carcinoma cells by a MUC1 aptamer-let-7i chimera.

The purpose of this study was to establish tumor tissue specific delivery of let-7i miRNA to reverse paclitaxel-induced chemoresistance. A chimera that combines MUC1 aptamer and let-7i miRNA was tested in OVCAR-3 ovarian cancer cells. Results demonstrated that the chimera can specifically be delivered into OVCAR-3 cells and the released let-7i significantly sensitized the role of paclitaxel in inhibiting cell proliferation, inducing cell apoptosis, and decreasing long-term cell survival. The chimera achieved reversal of chemoresistance through downregulation of cyclin D1, cyclin D2, Dicer 1, and PGRMC1 expressions. Our study indicated that this MUC1/let-7i chimera can specifically reverse chemoresistance to paclitaxel.

Phosphorylation of h1 calponin by PKC epsilon may contribute to facilitate the contraction of uterine myometrium in mice during pregnancy and labor.

BACKGROUND: The timely onset of powerful uterine contractions during parturition occurs through thick and thin filament interactions, similar to other smooth muscle tissues. Calponin is one of the thin filament proteins. Phosphorylation of calponin induced by PKC-epsilon can promote the contraction of vascular smooth muscle. While the mechanism by which calponin regulates the contraction of pregnant myometrium has rarely been explored. Here, we explore whether PKC-epsilon/h1 calponin pathway contribute to regulation of myometrial contractility and development of parturition. METHODS: We detected the expression of h1 calponin, phosphorylated h1 calponin, PKC-epsilon and phosphorylated PKC-epsilon in the different stages of mice during pregnancy and in labor by the method of western blot and recorded the contraction activity of myometrium strips at the 19th day during pregnancy with different treatments by the organ bath experiments. RESULTS: The level of the four proteins including h1 calponin, phosphorylated h1 calponin, PKC-epsilon and phosphorylated PKC-epsilon was significantly increased in pregnant mice myometrium as compared with that in nonpregnant mice. The ratios of phosphorylated h1 calponin/h1 calponin and phosphorylated PKC-epsilon/PKC-epsilon were reached the peak after the onset of labor in myometrium in the mice. After the treatment of more than 10(9-) mol/L Psi-RACK (PKC-epsilon activator), the contractility of myometrium strips from mice was reinforced and the level of phosphorylated h1 calponin increased at the same time which could be interrupted by the specific inhibitor of PKC-epsilon. Meanwhile, the change of the ratio of phosphorylated h1 calponin/h1 calponin was consistent with that of contraction force of mice myometrium strips. CONCLUSIONS: These data suggest that in mice myometrium, phosphorylation of h1 calponin induced by the PKC-epsilon might facilitate the contraction of uterine in labor and regulate pregnant myometrial contractility.

Alpha-1-antitrypsin acts as a preeclampsia-related protein: a proteomic study.

AIMS: To screen the preeclampsia-related protein by proteomics. METHODS: Proteomics was performed to identify differential protein expression profiles between normal full-term pregnancy, early-onset severe preeclampsia (ES-PE) or late-onset severe preeclampsia (LS-PE; n = 10 per group). Real-time quantitative PCR and immunohistochemistry were conducted to confirm the expression of α(1)-antitrypsin (α(1)-AT) in the decidual tissues of different subjects. ELISA was employed to detect the α(1)-AT content in the peripheral blood of 90 women (n = 30 per group). RESULTS: We successfully constructed two-dimensional electrophoresis maps of decidual tissues, and a total of 20 differentially expressed proteins were identified. The α(1)-AT expression was different among the three groups. The normal full-term pregnancy women expressed the most α(1)-AT, and the LS-PE women expressed the least amount of α(1)-AT. The difference in the α(1)-AT expression was consistent with the proteomics data. The peripheral α(1)-AT content was the highest in the normal full-term pregnancy group (1.85 ± 0.15 g/l), moderate in the ES-PE group (0.77 ± 0.14 g/l) and lowest in the LS-PE group (0.42 ± 0.07 g/l; p < 0.05). CONCLUSION: Using 2D PAGE, we identified twenty proteins with significantly altered expression in PE. These differentially expressed proteins include prevention protein, in which α(1)-AT is downregulated in PE.

[Expression of Rho-GDP dissociation inhibitor in the decidual tissues of preeclampsia patients and its clinical implication].

OBJECTIVE: To investigate the expression of Rho-GDI in the decidual tissues of patients preeclampsia and explore its clinical implication. METHODS: Real-time PCR, Western blotting and immunohistochemistry were used to detect the mRNA and protein expressions of Rho-GDI in the decidual tissues from 30 normal women with full-term pregnancy, 30 patients with early-onset severe preeclampsia and 30 with late-onset severe preeclampsia. RESULTS: Rho-GDI expression was found mainly on the cell membrane and in the cytoplasm and nuclei of the decidual cells, occasionally occurring in the stroma. Both the mRNA and protein expressions of Rho-GDI in the decidual tissues were significantly higher in the normal pregnancy group than in the two severe preeclampsia groups (P<0.05), and the patients with late-onset severe preeclampsia had the lowest expressions of Rho-GDI. CONCLUSION: The lowered expression of Rho-GDI in the deciduas might be involved in the pathogenesis and progression of preeclampsia.

Proportional changes of CD4+CD25+Foxp3+ regulatory T cells in maternal peripheral blood during pregnancy and labor at term and preterm.

PURPOSE: To evaluate the proportional changes of CD4+CD25+Foxp3+ regulatory T cells (Tregs) in maternal peripheral blood during pregnancy and labor at term and preterm. METHODS: Peripheral blood was collected from 20 non-pregnant controls and 139 pregnant women (60 at different gestational ages, 48 at term with or without labor, and 31 in threatened or actual preterm labor). CD4+CD25+Foxp3+ Tregs in peripheral blood samples were analyzed in peripheral blood samples by flow cytometry. Placentas from preterm women were examined for the presence of histological chorioamnionitis (HC). RESULTS: The percentage of circulating CD4+CD25+Foxp3+ Tregs was significantly increased in women during the first trimester compared with non-pregnant controls (P < 0.0001), peaking during the second trimester and then declining slightly in the third trimester. There was a significantly lower level of CD4+CD25+Foxp3+ Tregs in women with term labor than in those at term without labor (P < 0.0001). Women admitted in preterm labor had a lower proportion of CD4+CD25+Foxp3+ cells than those admitted with threatening preterm labor (P < 0.0001). Preterm women with evidence of HC had decreased proportion of CD4+CD25+Foxp3+ cells than those without HC in preterm deliveries (P=0.008). Moreover, the percentages of CD4+CD25+Foxp3+ cells in preterm subjects, irrespective of the HC status, were significantly diminished compared with women with normal pregnancy at the same gestational age (P < 0.0001). CONCLUSION: Our data reveal a marked elevation of peripheral blood CD4+CD25+Foxp3+ Tregs during early pregnancy, but a dramatic decline during labor, either at term or preterm, suggesting their involvement in the maintenance of pregnancy and the initiation of labor.

[Expression of Calponin-1 and Transgelin in human uterine smooth muscles in non-labor and labor situation].

OBJECTIVE: To investigate the expression of Calponin-1 and Transgenlin in the uterine smooth muscles during normal labor on-sets, and to evaluate their effect on initiating the normal labor. METHODS: A total of 14 uterine bodies and lower segments of human pregnancy were divided to a non-labor group (NIL) and a labor group(IL). Immunohistochemical technology and Western blot were used to determine the expression of Calponin-1 and Transgelin in the 2 groups. RESULTS: Immunohistochemical detection and Western blot showed that Calponin-1 protein in the uterine smooth muscle tissue of the body and the lower uterine segment of smooth muscle tissues had significant difference (P<0.05). The expression of Transgelin in the uterine body smooth muscle tissue in the IL was higher than that in the NIL(P<0.05). In the lower uterine segments of the smooth muscle, the expression of Transgelin was not significantly different in the 2 groups (P>0.05). CONCLUSION: Calponin-1 of the uterine smooth muscle and Transgelin of the uterine body smooth muscle may involve in the regulation of uterine smooth muscle contractility, which is closely related to child birth launch.

[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.

[Change of HIF-1alpha protein expression in the placenta bed and concentration of vWF in maternal peripheral blood of pre-eclampsia].

OBJECTIVE: To investigate the expression of HIF-1alpha protein in the placenta bed and the concentration of von Willrand factor (vWF) in maternal peripheral blood from pre-eclampsia and normal pregnancy, and to determine the effect of HIF-1alpha and vWF on the pathogenesis of pre-eclampsia. METHODS: Forty pre-eclampsia patients (20 mild and 20 severe) were recruited as 2 study groups, and another 20 normal pregnant women were served as a normal group.Western blot was used to detect the expression of HIF-1alpha protein in the placenta bed. ELISA was adopted to detect the concentration of vWF in the maternal peripheral blood.Correlation between HIF-1alpha protein expression and vWF level was analyzed by Spearman method. RESULTS: The expression of HIF-1alpha protein in the placenta bed was the highest in the severe pre-eclampsia patients among 3 groups, followed by the mild pre-eclampsia patients and the normal controls. There was significant difference among 3 groups (P<0.001). The concentration of vWF in the maternal peripheral blood was the highest in the severe pre-eclampsia patients, followed by the mild pre-eclampsia patients and the normal controls. There was significant difference among the 3 groups (P<0.05). Positive correlation was found between the expression of HIF-1alpha protein in the placenta bed and the concentration of vWF in the maternal peripheral blood in patients with pre-eclampsia (r1=0.65, P<0.001), and between the concentration of vWF in the maternal peripheral blood and the pathogenetic condition degree of pre-eclampsia (r(2)=0.61,P<0.001). CONCLUSION: HIF-1alpha in coordination with vWF may play an important role in the pathogenesis of pre-eclampsia.

[Effect of HPV16E6 on sensitivity of chemotherapy for cervical carcinoma in different p53 genotype cell lines].

OBJECTIVE: To investigate the effect of human papillomavirus types 16E6 on the sensitivity of chemotherapy for cervical carcinoma in different p53 genotype cell lines. METHODS: The apoptosis rates of each group were detected by AO/EB, immunofluorescence and Annexin V/PI stained methods. The expressions of protein HPV16E6 and p53(mt) after the treatments of different concentration of DDP were detected by Western blot. HPV16E6 mRNA in C33A, C33A-E6, C33A-P, and CaSki cell lines under different DDP treatments were detected by RT-PCR. RESULTS: AO/EB and Annexin V/PI stained tests showed that the apoptosis rates of C33A, C33A-E6, C33A-P, and CaSki cells were significantly increased when DDP concentration increased. Western blot showed that the HPV16E6 protein could be detected only in C33A-E6 and CaSki cell lines. The expression of HPV16E6 protein in C33A-E6 and CaSki cell lines gradually decreased and was hardly detected with increased dosage of DDP and the prolonged treatment time (P<0.01), and slightly increased in C33A-E6 and Caski cell lines without the treatment, but there was no significant difference between them (P>0.05). Protein p53(mt) persistently expressed in C33A-E6, C33A, and C33A-P cell lines following the increased dosage of DDP and the prolonged treatment time(P>0.05), while it couldn't be found in CaSki cell line. RT-PCR showed that without DDP intervention, there was no significant difference of HPV16E6 mRNA in C33A-E6 and CaSki cell lines within 24 h.The HPV16E6 mRNA in C33A-E6 cell line expressed much higher than that in CaSki (P<0.05), and HPV16E6 mRNA of 2 cell lines expressed much higher at 48 h than at 24 h (P<0.05).The expression of HPV16E6 mRNA in C33A-E6 and CaSki cell lines gradually decreased with the increased DDP and prolonged treatment time (P<0.01), while there was no significant difference between C33A-E6 and CaSki cell lines under the same DDP concentration (P>0.05). CONCLUSION: Effect of HPV16E6 on the sensitivity of chemotherapy for cervical carcinoma cell lines is not markedly related with the different p53 genotype forms(p53(mt)/p53wt ). HPV16E6 may affect the proliferation and sensitivity of chemotherapy in C33A cell line through other mechanism.

[Expression of annexin V in decidua tissues of preeclampsia patients].

OBJECTIVE: To investigate the expression of annexin V in the decidua tissues of preeclampsia patients and explore its clinical significance. METHODS: Real-time PCR, Western blotting and immunohistochemistry were employed to detect the mRNA and protein expressions of annexin V in the deciduas from 35 normal pregnant women at full term, 38 early onset severe preeclampsia patients and 33 late onset severe preeclampsia patients. RESULTS: Annexin V was found on the cell membrane and in the cytoplasm of the decidual cells and stroma. Both the mRNA and protein of annexin V expressions in the decidua tissues were significantly different between normal pregnancy group and early or late onset severe preeclampsia group (P<0.05), being the highest in normal pregnancy group and the lowest in early onset severe preeclampsia group. CONCLUSION: The low expression of annexin V in the deciduas might participate in the hypercoagulability state in preeclampsia patients.

[Expression of aspartyl-(asparaginyl) beta-hydroxylase in villi in patients with missed abortion].

OBJECTIVE: To determine the difference in aspartyl-(asparaginyl) beta-hydroxylase (AAH) expression level in villi between patients with missed abortion and normal women with early pregnancy, and to confirm the expression loci of AAH in villi. METHODS: A total of 50 patients of missed abortion were collected and categorized into a test group, which was subdivided into Group 1 and Group 2. Patients in Group 1 (n=20) were of confirmed etiological disorders while those in Group 2 (n=30) showed no obviously etiological clues. In addition, 20 women of early pregnancy with artificial abortion were categorized into a control group, whose embryos were sonographically confirmed alive before surgery. The 50 patients of missed abortion were also subdivided into a group within 4 weeks and a group over 4 weeks according to the time that the embryo stayed in utrine after death. Immunohistochemical technique and computer image analysis were used to detect the expression loci and the level of AAH in villi. RESULTS: AAH was expressed in the endochylema and nucleus of trephocyte both in missed abortion and normal early pregnancy. The expression level of AAH in villi of missed abortion was much lower than that of in villi of normal early pregnancy (P<0.05). The expression level had no difference between different groups of patients with missed abortion(P>0.05). CONCLUSION: Low expression of AAH in the endochylema and nucleus of trephocyte may play a role in patients with missed abortion.

[Construction of adenoviral vector encoding Calponin-1 siRNA and its effect on human myometrium cells in vitro].

OBJECTIVE: To investigate the effect of Calponin-1 suppression on human myometrium cells through adenovirus mediated siRNA. METHODS: Human uterine smooth muscle tissues were digested with enzymes, cultured and confirmed with immunocytochemistry. Adenovirus siRNA-Calponin-1 plasmid was transfected into primary cultured uterine smooth muscle cells in vitro. The expressions of Calponin-1 mRNA and protein were analyzed by RT-PCR and Western blot, respectively. RESULTS: The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid was successfully constructed, and Calponin-1 siRNA mediated by recombinant adenovirus resulted in markedly reduced expression of Calponin-1 mRNA and protein in human myometrium cells. The gray values of Calponin-1 mRNA in the uterine smooth muscle cells in the experimental, blank control, and empty vector groups were 316.3+/-39.2, 1048.5+/-126.4 and 1027.2+/-127.5, respectively. The gray values of Calponin-1 protein were 323.3+/-43.2, 1021.5+/-143.4, and 1019.2+/-144.5, respectively. The difference between the experimental group and the blank control group as well as the empty vector group was significant (P< 0.05). There was no significant difference between the empty vector group and the blank control group (P>0.05). CONCLUSION: The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid can inhibit the expression of Calponin-1 in human myometrium cells in vitro, which may be a useful approach to determine the role of Calponin-1 in delivery.

[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.

[Trophoblast cells invaing the placenta bed and change of spiral arteries and microvessels in pre-eclampsia].

OBJECTIVE: To investigate the invason of trophoblasts in the placenta bed and the change of spiral arteries and microvessels in pre-eclampsia and normal pregnancy. METHODS: Twenty cases of normal pregnancies, mild pre-eclampsia and severe pre-eclampsia were chosen as Group A, Group B, and Group C. HE staining and immunohistochemistry staining (SP method) were used to observe the depth and the density of trophoblasts invading the placenta bed and the change of spiral arteries and microvessels. RESULTS: The significant difference in the degree of invasion was in the superficial myometrial segment. Group C was the most superficial in the 3 groups (P<0.01). The density of trophoblasts which invaded the placenta bed in the lower half of the basal decidual segment and the myometrial segment showed us Group C was the lowest (P<0.01). There was statistical difference among the 3 groups (P<0.01). The average lumen area of the spiral arteries in the decidual segment and the superficial myometrial segment of the placenta bed was the smallest in Group C among the 3 groups(P<0.01) and there was statistical difference among the 3 groups (P<0.01). The spiral arteries were the thickest in Group C with statistical difference among the 3 groups (P<0.01). The physiological and pathological change of the spiral arteries was mainly in the superficial myometrial segment. The incidence rate of physiological changes in the spiral arteries was the lowest in Group C with statistical difference among the 3 groups (P<0.01). The incidence rate of pathological changes was the highest in Group C (P<0.01) and the normal group was the highest. There was significant difference among the 3 groups(P<0.01). There was positive correlation between the physiological change of the spiral arteries and the invaing degree of the trophoblasts (P<0.05), there was negative correlation between the pathological change of the spiral arteries and the invasion depth as well as the invasion density of the trophoblasts(P<0.05). There was negative correlation between the physiological change and the pathogenetic condition of pre-eclampsia(P<0.05)while there was positive correlation between the pathological change and the pathogenetic condition degree of pre-eclampsia(P<0.05). There was negative correlation between the invasion depth as well as density in uteruso superficial myometrial segment by trophoblast and the pathogenetic condition degree of pre-eclampsia(P<0.05). There was invasion trophoblast in 62.50% lumen wall of spiral arteries in uterus superficial myometrial segment of the placental bed in normal pregnancy while 27.5% was seen in severe pre-eclampsia. Microvascular density in the decidual segment and the superficial myometrial segment of the placenta bed in Group C was the lowest among the 3 groups with statistical difference (P<0.01). CONCLUSION: The invasion depth of the trophoblasts in pre-eclampsia was more superficial than normal pregnancy.The changes of the invasion of the trophoblasts and the pathological changes of the spiral arteries in the placenta bed mainly existed in the superficial myometrial segment which was closely related to the severity of the illness. That microvascular density in the placental bed of pre-eclampsia started to decrease from the basal decidual segment shows that the microvessel development in the placenta bed is impaired in pre-eclampsia.