Study on the expressions of PHD and HIF in placentas from normal pregnant women and patients with preeclampsia.

OBJECTIVE: To investigate the relationship between oxygen sensitivity of trophoblast and hypoxia in preeclamptic placenta by the study on the expressions of hypoxia-inducible factor prolyl 4-hydroxylase (PHD) and hypoxia-inducible factor (HIF) in placentas from normal pregnant women and patients with pre-eclampsia. METHODS: Subjects were chosen from the in-patients or the out-patients from May 2003 to May 2004. They were divided into 5 groups: early pregnancy group (EP), 13 cases; middle pregnancy group (MP), 9 cases; late pregnancy group (LP, or control group), 12 cases; preeclampsia (PE) group, 20 cases; gestational hypertension group (GH), 10 cases. The mRNA expressions of PHD-1 and -2 and -3 in placentas from all the subjects were assessed by in situ hybridization and Real-time PCR. The expressions of HIF-1α and -2α in placentas from different groups were assessed by immunohistochemistry and western blot. RESULTS: PHD-1,-2 and -3 mRNA were mainly expressed in cytoplasm of trophoblast, especially strongly expressed in extravillous trophoblast. During the progress of pregnancy, the expression of PHD-1 increased significantly (R=0.616, P<0.001). The PHD-1mRNA expression in placentas from PE group decreased significantly compared with that from control group, P<0.05. A significant direct correlation between the PHD-1 mRNA expression in placentas from PE group and their placenta weight was found (R=0.457, P<0.05). The HIF-2α, not the HIF-1α expression, from PE group was significantly higher than that from control group, P<0.01; The HIF-2α expression in trophoblast from PE was inversely correlated to the date of the onset of the disease (R=-0.730, P<0.01). CONCLUSIONS: PHD-1 played an important role in hypoxic response pathway of trophoblast through modulating the level of HIF-2α. The overly activated hypoxic response pathway of trophoblast in preeclamptic placenta, which is manifested as the result of HIF-2α over-expression, is the key point to hypoxic dysfunction of trophoblast.

Dysregulation of macrophage activation by decidual regulatory T cells in unexplained recurrent miscarriage patients.

CD4(+)CD25(+) T cells (Treg cells) and macrophages play roles in the maintenance of maternal-fetal immunological tolerance. Treg cells suppress the function of macrophages via mechanisms mediated by cell-cell contact and production of soluble factors. The purpose of this study was to investigate regulation of macrophages by Treg cells within decidua from patients with unexplained recurrent miscarriage (RM) and normal control women during early pregnancy. Treg cells and macrophages were isolated from deciduas of unexplained RM (n=15) and control women (n=15) by magnetic cell separation and co-cultured for six days. Regulation of macrophages by Treg cells was assessed in the presence and absence of neutralizing anti-TGFß antibodies and in transwell experiments. Expression of CD80, CD86, IL10, and IFNγ by macrophages was measured by flow cytometry or ELISA. Macrophage expression of CD80 and CD86 was higher in deciduas of unexplained RM patients compared with controls whereas the expression of IL10 was lower. There was no difference in the expression of IFNγ by macrophages between the two groups. Treg cells inhibited macrophage expression of CD80, CD86 and IFNγ and increased the expression of IL10. The regulatory effects of Treg cells were abrogated in the presence of neutralizing anti-TGFß antibodies or by transwell culture. The phenotype of macrophages therefore differed in unexplained RM patients compared with normal early pregnant subjects. Macrophage regulation by Treg cells was shown to be mediated by cell-cell contact and TGFß and this capacity was decreased in unexplained RM patients.

Pathogenesis, diagnosis, and treatment of recurrent spontaneous abortion with immune type.

Recurrent spontaneous abortion (RSA), defined as three or more consecutive pregnancy losses before 20 weeks of gestation, is difficult to treat in the clinical setting. It affects 1%-5% of women of reproductive age. In the investigations of immunopathogenesis, diagnosis, and treatment of RSA since the late 1980s, it was found that RSA was associated with abnormal maternal local or systemic immune response. The pathogenesis of autoimmune RSA was mainly associated with antiphospholipid antibody (APA), while that of alloimmune RSA was due to the disturbance of maternofetal immunological tolerance. Systemic etiological screening process and diagnosis systems of RSA with immune type were developed, and anticardiolipin (ACL or ACA) + anti-ß2-GP1 antibody combining multiple assays for effective diagnosis of RSA with autoimmune type was first established. According to the dynamic monitoring of clinical parameters before and during gestation, low-dose, short-course, and individual immunosuppressive therapy and lymphocyte immunotherapy for RSA with immune type were carried out. The outcomes of the offsprings of patients with RSA were followed up, and the safety and validity of the therapies were confirmed. The research achievement leads to great progress in the diagnosis and treatment of RSA in China.

Effect of antioxidants on amelioration of high-risk factors inducing hypertensive disorders in pregnancy.

BACKGROUND: This is a prospective clinical study based on a large sample gathered from multiple centers in China, subordinating to 10th Five-Year Plan of National Science & Technology Progression. We analyzed the high-risk factors inducing hypertensive disorders in pregnancy (HDP) and estimated the potential effect of anti-oxidants administration, including vitamin C (VC), vitamin E (VE) and Salvia Miltiorrhiza L (SML), a Chinese herb medicine, in amelioration of the high-risk factors in pregnancy. METHODS: From April 2005 to July 2006, 4814 pregnant women from 24 national wide cooperative hospitals were involved in this prospective research. The participants were randomly divided into two groups: 1607 cases were in anti-oxidants group with administration of vitamins and SML; 3207 cases were in control group without any medicine given. Every participant was under monitoring for the morbidity of HDP and the high-risk factors were investigated in HDP cases in each group. RESULTS: (1) The morbidity of HDP was 3.55% in anti-oxidants group vs. 4.18% in control group. No statistical difference existed between the two groups (P > 0.05). (2) In anti-oxidants group, the HDP morbidities among three subgroups: VC + VE + SML, VC + VE and SML only, were 5.51%, 3.05% and 5% respectively. It showed no statistical difference among three remedies (P > 0.05). (3) The related index of factors affecting HDP showed in intensity sequence as follows: family HDP history > profession > education level > age > body weight. The incidence of HDP in normal population was 3.51%, and the incidence of HDP in high-risk pregnant women (family HDP history, heavy physical labor, low education level (middle school and below), age ≥ 40, body mass index ≥ 24) was 5.84%, which was obviously higher than that in normal population (P < 0.01). In anti-oxidants group, the probability of HDP in women with high-risk factors was 3.81%, which was obviously lower than that in control group with high-risk factors at 7.14% (P < 0.01). (4) In control group, the morbidity of HDP in women with family HDP history (especially with sisters'), heavy physical labor, middle school and below, age ≥ 35 was: 50.00%, 15.22%, 6.33%, 26.28% and 5.75%, respectively, and that in anti-oxidants group was 0, 7.69%, 3.74%, 9.27% and 2.67%, respectively, which was obviously lower than that in control group. CONCLUSIONS: The high-risk factors prone to induce HDP included: family history of HDP, heavy physical labor, low education level, aging and obesity. No impressive effect of anti-oxidants application was found in preventing HDP in general population but the remedy demonstrated positive effect on preventing HDP in pregnant women with high-risk factors.

Pregnancy outcomes in women with heart disease.

BACKGROUND: As the Shanghai Obstetrical Cardiology Intensive Care Center, our hospital has accumulated a large number of clinical data of pregnant women with heart disease. This paper is a retrospective analysis of 1142 pregnancies in women with heart disease so as to evaluate the maternal and fetal outcomes of these patients. METHODS: A retrospective analysis was carried out for pregnancies in 1142 women with heart disease who delivered in Shanghai Obstetrical Cardiology Intensive Care Center between 1993 and 2007. RESULTS: In this study, main heart diseases in pregnancy were arrhythmia (n = 359, 31.4%), congenital heart disease (CHD; n = 291, 25.5%), and myocarditis and its sequelae (n = 284, 24.9%); based on the functional classification criteria of New York Heart Association (NYHA), more than half (n = 678, 59.4%) of patients were classified NYHA Class I; pregnant women in NHYA Class I-II (n = 951, 83.3%) commonly had arrhythmia, myocarditis and its sequelae, while those in NHYA Class III-IV (n = 191, 16.7%) mainly had CHD, rheumatic heart disease (RHD), cardiopathy induced by hypertensive disorders complicating pregnancy, and peripartum cardiomyopathy (PPCM). Cardiac failure occurred in 97 (8.5%) patients, and 8 (0.7%) maternal deaths and 12 (1.1%) perinatal deaths were reported in this study. Compared with those in NHYA Class I-II, women in NHYA Class III-IV had a significantly lower gestational age at birth (P < 0.05), lower birth weight (P < 0.01), and higher incidence of preterm delivery, small for gestational age and perinatal death (P < 0.01). The incidence of cardiac failure in pregnant women with cardiopathy induced by hypertensive disorders complicating pregnancy and PPCM was relatively high, with a rate of 80% and 52.2%, respectively. After cardiac operation, 131 (90.3%) women were in classified NHYA Class I-II and 14 (9.7%) in NHYA Class III-IV. CONCLUSIONS: Arrhythmia is the type of heart disease that has a highest incidence in patients with heart disease in pregnancy, while main types of heart disease that impair cardiac function are CHD and RHD; cardiac failure is more frequently caused by cardiopathy induced by hypertensive disorders complicating pregnancy and PPCM; impaired cardiac function increases perinatal morbidity; cardiac surgery before pregnancy could improve the cardiac function.

[A multi-center study to evaluate the dynamic changes of uterine artery and umbilical artery flow in a normal pregnancy and hypertensive disorders in pregnancy].

OBJECTIVE: To investigate the dynamic changes of uterine artery and umbilical artery in the first, second, and third trimester of normal pregnancy and hypertensive disorders in pregnancy (HDP). METHODS: A multi-center prospective study was conducted on 1098 women with normal singleton pregnancies at the first prenatal visit in the Second West China Hospital of Sichuan University, First Affiliated Hospital with Nanjing Medical University, Beijing Obstetrics and Gynecology Hospital Affiliated to Capital Medical University, Wuhan Union Hospital Affiliated to Medical School of Huazhong University of Science and Technology and Renji Hospital Affiliated to Medical School of Shanghai Jiao Tong University from April 2005 to July 2006, with the average age of (28.3 ± 3.3). The pulsatility indices (PI), resistance indices (RI) and systolic to diastolic velocity ratios (S/D) of uterine artery and umbilical artery were measured for all subjects in the first (10th-14th gestational weeks), second (20th-26th gestational weeks) and third trimester (30th-36th gestational weeks), respectively. In this longitudinal study, women who developed HDP were classified into HDP group, and the rest into normal pregnancy group. RESULTS: (1) Among the 1098 pregnant women, 44 developed HDP during the index pregnancy, including 20 gestational hypertension, 15 mild pre-eclampsia and 9 severe pre-eclampsia, giving an incidence of 4.17%, and the rest 1054 were normal until delivery. (2) In the normal pregnancy group, the RI, PI and S/D of uterine artery were decreased with the progress of pregnancy (RI: 0.64, 0.57, 0.50; PI: 1.24, 0.98, 0.80; S/D: 3.26, 2.58, 2.20; P < 0.01). However, the above indices showed an increasing trend with the increase of gestations in the HDP group (RI: 0.55, 0.67, 0.64; PI: 1.22, 1.36, 1.20; S/D: 3.18, 3.41, 3.05; P < 0.01). In the second and third trimester, the RI, PI and S/D of uterine artery in the HDP group were higher than those in the normal pregnancy group (P < 0.01). (3) In the normal pregnancy group, the RI, PI and S/D of the umbilical artery decreased from the second to the third trimester (RI: 0.71 and 0.58; PI: 1.16 and 0.87; S/D: 3.58 and 2.48; P < 0.01). However, no significant difference was found in the RI, PI and S/D value of umbilical artery in the second and third trimester between the normal and HDP group (RI: 0.71 and 0.63; PI: 1.20 and 0.95; S/D: 3.71 and 2.69; P > 0.05, respectively), despite the decreasing trend in the HDP group. CONCLUSIONS: In uncomplicated pregnancies, the blood flow resistance of uterine artery decreases and the end-diastolic blood flow of uterine artery increases with the progress of pregnancy. However, in pregnant women with HDP, the blood flow resistance of uterine artery increases significantly with the increase of gestations which shows significant difference to that in normal pregnancies. The blood flow resistance of umbilical artery decreases in both normal and HDP pregnant women with the increasing gestational age.

Role of cyclooxygenase-2 signaling pathway dysfunction in unexplained recurrent spontaneous abortion.

BACKGROUND: Experimental evidence indicates that cyclooxygenase-2 (COX-2) plays a critical role in blastocyst implantation; however, little is known of the role of COX-2 in unexplained recurrent spontaneous abortion (URSA). METHODS: We evaluated the expression level and potential signaling pathway of COX-2 in 30 cases of URSA who were excluded the abnormality of chromosomes, anatomy, endocrine, infectious, autoimmune diseases and in 30 normal pregnancies. RESULTS: The mRNA and the protein expression level of COX-2 in the URSA group (-0.238 +/- 0.848, 0.368 +/- 0.089, respectively) were significantly lower than that in the control group (1.943 +/- 3.845, 1.046 +/- 0.108, respectively) (both, P < 0.01). The expression of prostaglandins PGF(2a), PGD(2), PGE(2), and PGI(2), in the URSA group ((2326.0 +/- 295.6) pg/ml, (2164.0 +/- 240.5) pg/ml, (238.7 +/- 26.4) pg/ml, (2337.0 +/- 263.0) pg/ml, respectively) were significantly lower than that in the control group ((3450.0 +/- 421.7) pg/ml, (3174.0 +/- 415.6) pg/ml, (323.5 +/- 43.8) pg/ml, (3623.0 +/- 460.4) pg/ml, respectively) (P < 0.05). The mRNA expression level of PPARbeta and RXRalpha (0.859 +/- 0.653, -0.172 +/- 0.752, respectively) in URSA group was significantly lower than that in the control group (1.554 +/- 1.735, 0.777 +/- 2.482, respectively) (both P< 0.05). The mRNA and protein expression levels of vascular endothelial growth factor-A (VEGF-A) in the URSA group (2.010 +/- 1.522, 0.35 +/- 0.46) was significantly lower than that in the control group (4.569 +/- 2.430, 0.750 +/- 0.350) (both P < 0.05). CONCLUSIONS: COX-2 and the COX-2-derived PGI(2) signaling pathway possibly play an important role in successful embryo implantation, and their decreased expression may result in URSA. The decreased expression may influence the expression of VEGF-A which interferes with placental angiogenesis causing failure of embryo implantation, leading to spontaneous abortion.

The deregulation of regulatory T cells on interleukin-17-producing T helper cells in patients with unexplained early recurrent miscarriage.

BACKGROUND: CD4(+)CD25(+) regulatory T cells (Tregs) are important for the maintenance of immune homeostasis by virtue of their ability to control T-cell proliferation in the peripheral blood (PB). We recently demonstrated that the prevalence of Tregs is decreased, whereas that of Th17 cells is increased, in the PB and decidua samples of patients with unexplained recurrent miscarriage (RM). In this study, we investigated whether the cytokine production of Th17 cells can be suppressed by the Tregs and elucidated the mechanism by which Tregs exert this suppressive effect. METHODS: Flow cytometry was used to analyze the surface phenotype and cytokine production of Th17 cells in the PB of women with unexplained RM (n = 17) and healthy women in early stages of pregnancy who underwent elective abortion (n = 20). The suppressive ability of Tregs on Th17 cells was assessed in in vitro co-cultures and transwell experiments. The amount of secreted interleukin-17 (IL-17) in the supernatants was measured by enzyme-linked immunosorbent assay (ELISA). The inhibitory activity of transforming growth factor-ß (TGF-ß) and IL-10 on IL-17 expression in CD4(+) T cells was assessed using ELISA. RESULTS: The proportions of IL-17-positive CD4(+) T cells, CC chemokine receptor type 6 (CCR6)-positive CD4(+) T cells and CCR6 expression of IL-17-positive CD4(+) T cells were higher in the PB samples of patients with unexplained RM than in PB of healthy control subjects. In vitro, Tregs could inhibit the expression of IL-17; more Th17 cells were inhibited in the control group than in the unexplained RM group. High-dose TGF-ß inhibited the expression of IL-17, whereas IL-10 inhibited IL-17 expression in a dose-dependent manner. CONCLUSIONS: IL-17 expression can be inhibited by Tregs. The suppressive activity of Tregs on Th17 cells was decreased in patients with unexplained RM. The ability of Tregs to suppress cytokine secretion might be effected by a cell-cell contact. TGF-ß and IL-10 could inhibit the expression of IL-17.

Increased prevalence of T helper 17 (Th17) cells in peripheral blood and decidua in unexplained recurrent spontaneous abortion patients.

T helper 17 (Th17) cells and regulatory T (Treg) cells are two distinct subsets of CD4(+) T cells which have opposite effects on inflammation, autoimmunity and immunological rejection of foreign tissue. Treg cells have been shown to be important in maintaining materno-fetal tolerance, but the role of Th17 cells in human pregnancy and pathological pregnancy, especially in relation to unexplained recurrent spontaneous abortion (RSA), has not been investigated. In this study, we showed that the proportion of Th17 cells in the peripheral blood and decidua was significantly higher in unexplained RSA patients compared to normal, early pregnant women. Meanwhile, there was an inverse relationship between Th17 cells and Treg cells in the peripheral blood lymphocytes (PBL) and decidua in unexplained RSA. The expression of Th17 related factors, IL-17, IL-23 and retinoid orphan nuclear receptor (RORC), in PBL and decidua in unexplained RSA patients was significantly higher than normal, early pregnant women. This study is the first to define the occurrence of Th17 cells in unexplained RSA patients and in normal, early pregnant women. We suggest that these highly pro-inflammatory cells contribute to unexplained RSA, and the balance between Th17 cells and Treg cells may be critical to pregnancy outcomes.

[Multi-center study of motherwort injection to prevent postpartum hemorrhage after caesarian section].

OBJECTIVE: To evaluate the efficacy and safety of motherwort (herba leonuri/leonurus heterophyllus sweet) injection for preventing postpartum hemorrhage after caesarian section. METHODS: The prospective study was designed as a randomized and single blind multi-center research matched with positive agent as controls from Apr 2007 to Aug 2007. 440 women underwent caesarian section (CS) indicated by obstetric factors were enrolled from 15 teaching hospitals in China and assigned into three groups: group of motherwort: 147 cases were administered by motherwort 40 mg uterine injection during CS and 20 mg intramuscular injection per 12 hours 3 times after CS; group of motherwort+oxytocin: 144 cases were administered by motherwort 40 mg and oxytocin 10 U uterine injection during CS and motherwort 20 mg intramuscular injection per 12 hours 3 times after CS and group of oxytocin: 149 cases were administered by oxytocin 10 U uterine injection and oxytocin 10 U+5% glucose 500 ml intravenously injection during operation and oxytocin 10 U intramuscular injection per 12 hours 3 times after CS. The following clinical parameter were collected and analyzed: (1) The amount of blood loss during operation, at 2, 6, 12, 24, 48 hours after operation. (2) The total amount of blood loss in 24 hours after CS and the incidence of postpartum hemorrhage. (3) The change of level of hemoglobin (Hb) and counting of red blood cell (RBC) from prepartum to postpartum. (4) Adverse reaction. RESULTS: (1) The mean amount of blood loss during operation were (368+/-258) ml in group of motherwort, (255+/-114) ml in group of motherwort+oxytocin and (269+/-141) ml in group of oxytocin, which exhibited significant difference among three groups (P<0.01). Meanwhile, no statistical different amount of blood loss among three groups were observed at 2, 6, 12, 24, 48 hours after CS. (2) The amount of blood loss of postpartum at 24 hours were (480+/-276) ml in group of motherwort, (361+/-179) ml in group of motherwort+oxytocin, (381+/-179) ml in group of oxytocin, which showed significant difference among 3 groups (P<0.01). (3) The incidence of postpartum hemorrhage were 32.0% (47/147) in group of motherwort, 11.1% (16/144) in group of motherwort+oxytocin, and 18.8% in (28/149) in group of oxytocin. When comparing the lowest rate of postpartum blood loss in group of motherwort+oxytocin and the highest rate in group of motherwort, it displayed statistical difference (P<0.01). (4) The decreased level of RBC and Hb were shown that RBC (0.3+/-0.5)x10(12)/L and Hb (9+/-13) g/L in group of motherwort, RBC (0.2+/-0.4)x10(12)/L and Hb (6+/-10) g/L in group of motherwort+oxytocin and RBC (0.2+/-0.4)x10(12)/L and Hb (7+/-30) g/L in group of oxytocin respectively. However, the comparison of different value of RBC and Hb in group of oxytocin and motherwort+oxytocin showed significant difference (P<0.05). (5) Two cases with allery reaction was observed. CONCLUSION: It is safe and efficacious that combined use of motherwort injection and oxytocin was to prevent postpartum hemorrhage during or after caesarian section.

[Regulative effects of regulatory T cells on dendric cells in peripheral blood and deciduas from unexplained recurrent spontaneous abortion patients].

OBJECTIVE: To study the effect of CD(4)(+)CD(25)(+) regulatory T(Tr) cells on dendric cells (DC) in peripheral blood and deciduas from unexplained recurrent spontaneous abortion (URSA) patients. METHODS: Four URSA patients (abortion group) and 4 normal early pregnant women (control group) were enrolled in this study. Tr cells and DC in the peripheral blood and deciduas were isolated using Ficoll density gradient centrifugation and magnetic cell sorting (MACS). DC were cultured alone (DC alone) or in combination with Tr cells (DC + Tr) for 6 days, during which the release of interferon (IFN)-gamma and interleukin (IL)-10 in the medium was subsequently measured by enzyme linked immunoadsorbent assay (ELISA). RESULTS: (1) Peripheral blood: there was no significant difference in IFN-gamma level between DC alone (23.2 +/- 0.7) ng/L and DC + Tr (22.5 +/- 3.0) ng/L in abortion group (P > 0.05). The similar level of IL-10 was observed between DC alone (37 +/- 7) ng/L and DC + Tr (35 +/- 4) ng/L in abortion group (P > 0.05). IL-10 level, but not IFN-gamma, was significantly higher in DC alone (54 +/- 20) ng/L than that in DC + Tr (36 +/- 9) ng/L in control group (P < 0.01). (2) Deciduas: there was no significant difference in IFN-gamma level between DC alone (23.4 +/- 2.6) ng/L and DC + Tr (24.4 +/- 2.5) ng/L in abortion group (P > 0.05). Moreover, Similar IL-10 level was found between DC alone (28 +/- 7) ng/L and DC + Tr (25 +/- 5) ng/L in abortion group (P > 0.05). IFN-gamma level in DC alone (30.7 +/- 4.6) ng/L was significantly higher than that in DC + Tr (22.6 +/- 3.8) ng/L in control group (P < 0.01); whereas IL-10 level was much lower in DC alone (27 +/- 6) ng/L than that in DC + Tr (31 +/- 9) ng/L in control group (P < 0.05). CONCLUSION: The decreasing of immunosuppressive function of Tr cell of URSA patients affect its regulation on DC, resulting in imbalance of Th1/Th2 and abnormality of maternal-fetal immuno-tolerence in URSA.

Relationship between expression of chemokine receptors CCR3, CCR5 and CXCR3 on CD4(+) T cells and spontaneous abortion in mice.

BACKGROUND: Previous studies have shown that local immune cells in the feto-maternal interface are recruited from peripheral blood, and that chemokines and their receptors play an initial and key role in this recruitment process. In this study, we aimed to determine whether spontaneous abortion is associated with the expression of chemokine receptors CCR3, CCR5, and CXCR3 on CD4(+) T cells. METHODS: Peripheral blood, spleen, and thymus were collected from the spontaneous abortion mouse model CBA/JxDBA/2 (SA group, n = 14), the normal pregnant mouse model CBA/JxBALB/c (NP group, n = 13), and normal non-pregnant CBA/J mice (NNP group, n = 11). The number of chemokine receptors CCR3, CCR5, and CXCR3 expressed on CD4(+) T cells was measured by double-label flow cytometry (FCM) method. RESULTS: In peripheral blood, the SA group had significantly lower CCR3 expression (P < 0.01) and higher CCR5 and CXCR3 expression (P < 0.01) on CD4(+) T cells than did the NP group. But comparing these chemokines between the SA and NNP groups, there was no significant difference (P > 0.05). In spleen, the SA group expressed significantly lower CCR3 expression (P < 0.01) and higher CCR5 and CXCR3 expression (P < 0.05) on CD4(+) T cells than did the NP group. When compared with the NNP group, the SA group had significantly higher CCR3 expression (P < 0.01), but was not statistically different with regards to the other two chemokines (P > 0.05). In thymus, the SA group had significantly lower CCR3 expression (P < 0.05) and higher CXCR3 expression (P < 0.05) on CD4(+) T cells than the NP group, with no significant difference in CCR5 expression (P > 0.05). Compared with the NNP group, the SA group had higher CCR3 expression (P < 0.01), but there was no statistical difference in CXCR3 and CCR5 expression (P > 0.05) between the two groups. CONCLUSION: The abnormal expression of CCR3, CCR5 and CXCR3 on CD4(+) T cells may play an important role in the pathogenesis of spontaneous abortion.

Coagulation and fibrinolysis related cytokine imbalance in preeclampsia: the role of placental trophoblasts.

OBJECTIVE: Cytokine imbalance might have a pivotal role in hypercoagulability seen in preeclampsia. Our objective was to determine the relationship of blood coagulation related factors in placental tissue and peripheral blood in preeclamptic and normal pregnancies. METHODS: We compared mRNA and protein levels of tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1), and tissue factor (TF) in the placenta of normal and preeclamptic pregnancies. Placental and peripheral blood t-PA and PAI-1 levels were examined. Trophoblasts were used to study the effects of hypoxia, hypoxia-reperfusion, and inflammatory cytokines on t-PA, PAI-1, tissue factor pathway inhibitor (TFPI), and TF. RESULTS: PAI-1 and TF mRNA and protein levels were higher in placental tissue of preeclamptic pregnancies and in the peripheral blood of patients with preeclampsia. mRNA and protein secretion of TF, TFPI, PAI-1, but not t-PA, was increased in trophoblast cell culture under hypoxia and hypoxia-reoxygenation. Cell cultures with high levels of tumor necrosis factor-alpha (TNF-alpha) exhibited increased expression and secretion of TF and PAI-1, decreased TFPI, and no significant change of t-PA. CONCLUSIONS: Imbalanced synthesis of t-PA, PAI-1, TFPI, and TF in trophoblasts may contribute to hypercoagulability in patients with preeclampsia.

Effects of letrozole and clomiphene citrate on the expression of HOXA10 and integrin alpha v beta 3 in uterine epithelium of rats.

OBJECTIVE: To compare the effects of letrozole and clomiphene citrate on the expression of HOXA10 and integrin alpha(v)beta(3) in the uterine epithelium in rats. DESIGN: Controlled prospective study. SETTING: Teaching hospital and university research laboratory. ANIMAL(S): Sixty sexually mature female Wistar-Albino rats that were 20 weeks of age. INTERVENTION(S): Letrozole, 5 mg/kg of body weight daily (20 rats); clomiphene citrate, 100 microg/kg daily (20 rats); and saline solution, 2 mL/day (20 rats). After 2 days, rats were killed and hysterectomized. MAIN OUTCOME MEASURE(S): Expression of integrin alpha(v)beta(3) and HOXA10. RESULT(S): The expression of integrin alpha(v)beta(3) in the clomiphene citrate group was statistically significantly lower than in the letrozole and saline solution groups. The expression of HOXA10 was statistically significantly higher in the saline solution group than in the letrozole group, and the letrozole group showed a statistically significantly higher expression of HOXA10 compared with the clomiphene citrate group. CONCLUSION(S): In rats, letrozole affects the expression of HOXA10 in uterine epithelium but has no effect on the expression of integrin alpha(v)beta(3), which suggests that clomiphene suppresses endometrial receptivity more than letrozole. In the future, letrozole may be an appropriate drug for ovulation induction.

CXCL12 enhances exogenous CD4+CD25+ T cell migration and prevents embryo loss in non-obese diabetic mice.

OBJECTIVE: To investigate the possible role of CXCL12 in the migration of regulatory T (Treg) cells. DESIGN: Animal model-based study. SETTING: Academic. ANIMAL(S): Pregnant non-obese diabetic (NOD) mice were compared with non-immunodeficient mice. INTERVENTION(S): In vivo and in vitro CXCL12 induction. MAIN OUTCOME MEASURE(S): Flow cytometric analysis and Treg cell migratory assay. RESULT(S): A significantly high percentage of spontaneous embryo resorption was observed in both syngeneic and allogeneic pregnant NOD mice. The percentage of embryo loss in allogeneic pregnant NOD mice was significantly decreased by treatment with Treg cells and CXCL12 injection; however, no such effect was observed in syngeneic pregnant NOD mice. In addition, the migration of Treg cells induced by CXCL12 was confirmed by both in vitro and in vivo migratory assays. CXCR4, the specific receptor for CXCL12, was expressed more intensively on Treg cells than on non-Treg CD3(+) T cells, whereas CXCL12 was dominantly expressed in cytokeratin 7(+) trophoblast cells at an early stage of gestation, and its expression reduced gradually during pregnancy. CONCLUSION(S): The higher level of embryo loss in allogeneic pregnant NOD mice may be due to the lack of Treg cells. CXCL12 can cause CXCR4(+) Treg cells to migrate into the pregnant uterus and establish a beneficial microenvironment for the fetus.

The investigation on the value of repeat and combination test of ACA and anti-beta2-GPI antibody in women with recurrent spontaneous abortion.

PROBLEM: In order to investigate the value of anticardiolipin antibodies (ACA) and anti-beta2-GPI antibodies detection in screening autoimmune type recurrent spontaneous abortion and its clinic application in antiphospholipid syndrome diagnosis, we adopt repeat combined ACA and anti-beta2-GPI antibodies detection in this study. METHOD OF STUDY: Sera were collected from patients and work-up was done for detection of ACA and anti-beta2-GPI antibodies by enzyme-linked immunosorbent assay (ELISA). The work-up was done for detection of antibodies once in every 6 weeks for 14 times consecutively. RESULTS: The repeated and combined detection of ACA and anti-beta2-GPI antibodies detection could raise the positivity rate up to 21.8% (P < 0.05) in comparison with positive for ACA alone (14.1%), positive for anti-beta2-GPI alone (3.1%), and concurrently positive for both ACA and anti-beta2-GPI antibodies (4.6%). In 91 confirmed positive antiphospholipid antibodies (APA) patients, with more frequent screening for ACA and anti-beta2-GPI antibodies, more patients with APA were found. The positive rate of five and more screenings was over 81.32%, which was statistically significant (P < 0.05), in comparison with that of four or less screenings (68.13%). CONCLUSION: Our data implied that it would be appropriate to take over five or more screenings of combined ACA and anti-beta2-GPI antibodies detection in suspect patients to facilitate the positive diagnostic rate for autoimmune type RSA.

[Changes in proportion of decidua and peripheral blood CD(4)(+)CD(25)(+) regulatory T cells in unexplained recurrent spontaneous abortion patients].

OBJECTIVE: To study the changes in the percentage of CD(4)(+)CD(25)(+) regulatory T (Tr) cells in peripheral blood and deciduas in unexplained recurrent spontaneous abortion (URSA) patients, normal non-pregnant and pregnant women respectively. METHODS: The percentage of CD(4)(+)CD(25)(+) Tr cells in deciduas and peripheral blood from 25 URSA patients, 22 normal non-pregnant (NNP) women, and 34 normal early pregnant (NP) women were measured by double-staining followed by flow cytometric analysis. RESULTS: (1) The percentage of CD(4)(+)CD(25)(bright) T cells in peripheral blood in both URSA and NP [(1.55 +/- 0.77)%, (2.65 +/- 1.10)%, respectively] women were increased significantly than that in NNP women [(0.39 +/- 0.14)%, P < 0.05]. The percentage of CD(4)(+)CD(25)(bright) T cells in peripheral blood in URSA women was significantly lower than that in NP women (P < 0.05). (2) The percentage of CD(4)(+)CD(25)(bright) T cells in decidua in URSA women was significantly lower than that in NP women [(0.59 +/- 0.23)%, (1.24 +/- 0.55)%, respectively, P < 0.01]. There was no significant difference in the percentage of CD(4)(+)CD(25)(dim) T cells in decidua between URSA women and NP women [(4.23 +/- 1.52)%, (3.75 +/- 1.88)%, respectively, P > 0.05]. (3) The proportion of CD(4)(+)CD(25)(bright)/CD(4)(+) cells in deciduas was significantly higher than that in peripheral blood in NP women [(13.10 +/- 10.25)%, (5.59 +/- 2.62)%, respectively, P < 0.05]. However, a significant difference in the proportion of CD(4)(+)CD(25)(bright)/CD(4)(+) between decidua and peripheral blood was not found in URSA patients [(5.16 +/- 2.83)%, (4.64 +/- 2.07)%, respectively, P > 0.05)]. CONCLUSIONS: The number of CD(4)(+)CD(25)(+) Tr cells is increased in normal pregnancy and decreased in URSA. Therefore, CD(4)(+)CD(25)(+) Tr cells may play an important role in maintaining maternal-fetal tolerance and may be involved in the pathogenesis of URSA.

[The skewed usage of T cell receptor beta variable chain at the maternal-fetal interface of women with unexplained recurrent spontaneous abortion].

OBJECTIVE: To investigate T cell receptor (TCR) variable beta (BV) chain usage at the maternal-fetal interface and explore the relationship between the skewed TCR BV usage and unexplained recurrent spontaneous abortion (RSA). METHODS: Eighteen cases with unexplained RSA, together with matched 41 women with normal pregnancies in first trimester from Renji Hospital, Shanghai Jiao Tong University were studied. A high-resolution spectrum typing analysis of complementarity-determining region 3 (CDR3) was used to detect and compare the degree and frequency of TCR BV family expression in deciduas between RSA patients and normal controls. RESULTS: (1) The expression degree of BV19 (0.029 +/- 0.031 vs. 0.013 +/- 0.010, P = 0.038) in RSA group showed a higher usage, while BV5.2 (0.040 +/- 0.035 vs. 0.067 +/- 0.052, P = 0.046) showed a significantly lower usage when compared with normal controls. No significant difference in the expression of the other TCR BV families between RSA and controls were observed (P > 0.05). (2) TCR BV2, 3, 6, and 7 were the four most common BV families in deciduas of patients with RSA and normal controls, whose frequencies were all more than 50%. In RSA group, higher frequencies of BV15 (33.3% vs. 7.3%, P = 0.018), BV19 (38.9% vs. 14.6%, P = 0.049) and BV20 (33.3% vs. 7.3%, P = 0.018) were observed; meanuhile lower frequencies of BV4 (33.3% vs. 65.9%, P = 0.026) and BV7 (66.7% vs.92.7%, P = 0.018) distributions were observed. The other TCR BV families did not display significantly different freqencies of distribution (P > 0.05). CONCLUSIONS: It is suggested that a significant skewed TCR BV family occurs at the maternal-fetal interface in patients who undergo abortion. The specific skewed usages of TCR BV might be associated with the susceptibility to unexplained pregnancy loss.

[Study on the expression of transcription factor GATA-3 and T-bet mRNA in decidua of women with unexplained recurrent spontaneous abortion].

OBJECTIVE: To investigate the roles of transcription factor GATA-3 and T-bet at the fetal-maternal interface in the pathogenesis of unexplained recurrent spontaneous abortion (URSA). METHODS: The expression of GATA-3 and T-bet mRNA was examined by in situ hybridization. Decidua was obtained from 20 women with URSA and 20 normal pregnant (NP) women. RESULTS: (1) The number of GATA-3 positive cells per high power field in women with URSA (25 +/- 16) was significantly lower than those in NP women (38 +/- 16) (P < 0.05). (2) The number of T-bet positive cells per high power field in women with URSA (59 +/- 17) was significantly higher than those in NP women (46 +/- 18) (P < 0.05). (3) Numbers of cells expressing GATA-3 mRNA correlated significantly with T-bet mRNA positive cell numbers in decidua (r = -0.55, P < 0.01). CONCLUSIONS: The results demonstrate that upregulated expression of T-bet and downregulated expression of GATA-3 are found in the decidua of women with URSA. Imbalance of GATA-3/T-bet transcription factors might interfere with type 1 helper T cell (Th1)/type 2 helper T cell (Th2) differentiation at the fetal-maternal interface and cause URSA.