Direct antiglobulin test-negative autoimmune hemolytic anemia in a patient with ß-thalassemia minor during pregnancy: A case report.

BACKGROUND: Severe refractory anemia during pregnancy can cause serious maternal and fetal complications. If the cause cannot be identified in time and accurately, blind symptomatic support treatment may cause serious economic burden. Thalassemia minor pregnancy is commonly considered uneventful, and the condition of anemia rarely progresses during pregnancy. Autoimmune hemolytic anemia (AIHA) is rare during pregnancy with no exact incidence available. CASE SUMMARY: We report the case of a 30-year-old ß-thalassemia minor multiparous patient experiencing severe refractory anemia throughout pregnancy. We monitored the patient closely, carried out a full differential diagnosis, made a diagnosis of direct antiglobulin test-negative AIHA, and treated her with prednisone and intravenous immunoglobulin. The patient gave birth to a healthy full-term baby. CONCLUSION: Coombs-negative AIHA should be suspected in cases of severe hemolytic anemia in pregnant patients with and without other hematological diseases.

GNG7 silencing promotes the proliferation and differentiation of placental cytotrophoblasts in preeclampsia rats through activation of the mTOR signaling pathway.

Preeclampsia (PE) is a pathological condition that manifests during pregnancy as the occurrence of an abnormal urine protein level and increased blood pressure due to inadequate cytotrophoblast invasion. To elucidate the mechanism underlying PE, the present study primarily focused on the regulatory effects and mechanism of the G protein γ 7 (GNG7) on placental cytotrophoblasts in a rat PE model. Initially, the PE model was established with 45 specific pathogen­free Sprague­Dawley rats (30 females and 15 males). The expression patterns of GNG7, 4E­binding protein 1 (4E­BP1), phosphoprotein 70 ribosomal protein S6 kinase (p70S6K) and mammalian target of rapamycin (mTOR) were examined in the PE rats. Placental cytotrophoblasts isolated from normal and PE rats were treated with a small interfering RNA against GNG7, mTOR signaling pathway activator (HIV­1 Tat) or inhibitor (rapamycin). Following treatment, cell proliferation, differentiation and apoptosis were evaluated, and mTOR signaling pathway­related factors (4E­BP1, p70S6K and mTOR), cell proliferation­related factors (vascular endothelial growth factor and transforming growth factor­ß1), differentiation­related factors [activator protein­2 (AP­2)α and AP­2γ], and apoptosis­related factors [B­cell lymphoma 2 (Bcl­2) and Bcl­2­associated X protein] were determined. Finally, soluble fms­like tyrosine kinase 1 (sFlt­1) and soluble endoglin (sEng) levels were measured via enzyme­linked immunosorbent assay. Initially, the mTOR signaling pathway was inactivated in the placental tissues and cytotrophoblasts in the PE rats. Silencing GNG7 reduced the levels of sFlt­1 and sEng and activated the mTOR signaling pathway. Silencing of GNG7 or activation of the mTOR signaling pathway enhanced cell proliferation and differentiation, but inhibited the apoptosis of placental cytotrophoblasts in the PE rats. Taken together, the results showed that GNG7 silencing repressed apoptosis and enhanced the proliferation and differentiation of placental cytotrophoblasts in PE rats through activation of the mTOR signaling pathway.

Akt-mTOR Signaling Mediates Abnormalities in the Proliferation and Apoptosis of Ovarian Granulosa Cells in Patients with Polycystic Ovary Syndrome.

BACKGROUND/AIMS: Abnormal apoptosis of granulosa cells (GCs) is thought to involve in the pathogenesis of polycystic ovary syndrome (PCOS); however, the associated cellular and molecular mechanisms remain unclear. METHODS: Primary GCs were obtained from healthy women and women with PCOS. The cell proliferation and apoptosis were analyzed in insulin-stimulated and insulin receptor gene (INSR) siRNA-transfected GCs. The protein expression of Akt-mTOR-S6K1 signal molecules was measured by Western blot. RESULTS: This study showed that 1 nM of insulin significantly stimulated cell proliferation, induced cell apoptosis, and decreased the telomerase activity in GCs from both the healthy women and PCOS patients (p < 0.001), but silencing of INSR expression blocked the effects of insulin. Insulin induced significantly more apoptosis in GCs from PCOS patients than from healthy women (p < 0.01). Insulin significantly increased the ratio of p-Akt/Akt, the expression of mTOR protein, and the ratio of p-S6K1/S6K1 in GCs from normal control than in cells from PCOS patients (p < 0.001). CONCLUSION: Insulin-induced apoptosis of GCs, less activation of Akt-mTOR signaling, and reduction of telomerase activity may be associated with the pathogenesis of PCOS.

miR-148a-mediated estrogen-induced cholestasis in intrahepatic cholestasis of pregnancy: Role of PXR/MRP3.

Intrahepatic cholestasis of pregnancy (ICP) is an idiopathic liver disease while the biochemical characteristic is the elevated level of total bile acid (TBA). The present study investigated whether miR-148a mediates the induced effect of estrogen on the development of ICP and the proper mechanism: PXR/MRP3 signal pathway. mRNA expression was detected by qPCR, protein expression was detected by western blotting, the concentration of estrogen and TBA were detected by reagent kit respectively. In the cinical research, it was found that miR-148a expression was positive related with the concentration of TBA in the serum of ICP patients. In in vitro research, estradiol (500 nmol/L, 12 h) significantly upregulated miR-148a expression and LV-148a-siRNA inhibited the function of estradiol (500 nmol/L, 48 h) on TBA secretion. In addition, gene silence of miR-148a upregulated PXR expression which was inhibited by estradiol in LO2 cells. Pretreatment of rifampin (10 µmol/L), the agonist of PXR alleviated the TBA secretion induced by estradiol (500 nmol/L, 48 h). miR-148a-siRNA and PXR had a synergistic action on TBA secretion of LO2. Both of miR-148a-siRNA and rifampin (10 µmol/L) inhibited the upregulated effect of estradiol on MRP3 expression. This research has demonstrated that miR-148a may be involved in the induction of estrogen on ICP via PXR signal pathway, and MRP3 may be involved.

Nexrutine inhibits cancer cell growth as a consequence of mitochondrial damage and mitophagy.

BACKGROUND/AIMS: Nexrutine is an herbal extract of Phellodendron amurense and has been used as nutrient supplement in China as well as America. Potential protection effect of Nexrutine has been reported. METHODS: To investigate the mechanism of Nexrutine, we used the HeLa, U2OS and HCT116 as a model. Based on the acidification of cell culture media, we examined the lactate, mitochondria damage as well as mitophagy status by corresponding assay. RESULTS: Our data suggest that Nexrutine alters the cellular glucose metabolism to promote lactate production. This effect is caused by mitochondrial damage, not an alteration to lactate dehydrogenase activity. As a result of the mitochondrial damage, cell proliferation was inhibited and was associated with an elevation in p21/p27 proteins, which are both important cell cycle inhibitors. As another consequence of the mitochondrial damage, mitophagy was highly activated in Nexrutine-treated cells in a dose-dependent manner. When the autophagy pathway was blocked by siRNAs against BECN1 or ATG7, the growth inhibition caused by Nexrutine was reversed. CONCLUSION: Our study revealed that autophagy plays an important role in the inhibition of cancer cell proliferation by Nexrutine.

The impact of intrahepatic cholestasis of pregnancy with hepatitis B virus infection on perinatal outcomes.

INTRODUCTION: To investigate the impact of intrahepatic cholestasis of pregnancy (ICP) with hepatitis B virus (HBV) infection on perinatal outcomes. METHODS: In the study, 200 pregnant women were divided into four groups, including 50 cases with ICP and HBV infection, 50 cases with ICP, 50 cases with HBV infection, and 50 healthy pregnancies. The delivery process and perinatal outcomes were analyzed among different groups. RESULTS: When compared to the healthy pregnancy group, significantly increased rates of premature rupture of membranes, meconium-stained amniotic fluid, and cesarean section were observed in cases of ICP, HBV infection, or ICP patients with HBV (P<0.05). Specifically, the rates of HBV infection in the newborn, fetal distress, neonatal asphyxia, and birth defects in the newborn, and infant Apgar scores were higher in ICP pregnancies with HBV (56%, 48%, 16%, and 48%, respectively) than in the other groups (P<0.05). CONCLUSION: ICP combined with HBV infection has a clear influence on perinatal infant outcomes.

Two cases of acute aortic dissection following preeclampsia in non-Marfan patients.

Aortic dissection accompanying with preeclampsia during pregnancy can be lethal to both the mother and the fetus and carries a high mortality. Of the 2 preeclampsia patients with aortic dissection, one was Type B aortic dissection, occurring in postpartum period. The patient was treated medically and underwent catheter-based stent-graft treatment with fenestration technique. Another patient was Type A acute dissection, occurring in the third trimester. This patient was undiagnosed and both died. Although extremely rare, aortic dissection might be a possibility in preeclampsia pregnant women, the differential diagnosis of chest and/or epigastric pain in preeclampia patient should be thoroughly investigated and treated.

[Relationship between total bile acid concentration and fetal pulmonary surfactant in intrahepatic cholestasis of pregnancy].

OBJECTIVE: To explore the relationship between total bile acid (TBA) concentration and fetal pulmonary surfactant in intrahepatic cholestasis of pregnancy (ICP). METHODS: Fifty five patients with ICP (ICP group) who received cesarean section from April 2008 to February 2010 in Second Xiangya Hospital, Central South University, were recruited. The general conditions of the neonates within 7 days after birth in ICP group were recorded. Those with fetal distress, neonatal asphyxia, or neonatal respiratory distress syndrome were referred as pathological neonates, others were referred as normal neonates. Over the same period, 23 healthy gravidas were recruited as control group. Enzymatic method was used to detect the TBA concentrations in maternal blood, cord blood and amniotic fluid. ELISA was employed to measure the urfactant protein A (SP-A) concentration in cord blood. High performance liquid chromatography system was used to detect the concentrations of phosphatidylcholine (PC), phosphatidylinositol (PI), lysophosphatidylcholine (LPC), and sphingomyelin (SM) in amniotic fluid. RESULTS: (1) The concentrations of TBA in maternal blood, cord blood and amniotic fluid were (30.1 ± 7.9), (9.3 ± 3.3) and (4.4 ± 1.5) mmol/L in ICP group, (4.8 ± 2.2), (4.9 ± 0.9) and (1.4 ± 1.1) mmol/L in control group, respectively. The differences between the two groups were significant (P < 0.05). (2) The SP-A concentration in cord blood in ICP group was (29.5 ± 6.4) µg/L, significantly higher than that in control group, which was (22.6 ± 7.4) µg/L (P < 0.05). (3) There were 20 pathological neonates and 35 normal neonates in ICP group. In pathological neonates, the concentrations of TBA and SP-A in cord blood were (10.9 ± 2.2) mmol/L, (37.0 ± 5.9) µg/L, respectively; and were (8.0 ± 2.8) mmol/L, (26.7 ± 4.8) µg/L in normal neonates. The differences were significant (P < 0.05). (4) There was a positive correlation between TBA concentration in cord blood and in maternal blood (r(1) = 0.706, P < 0.05). The TBA concentration in cord blood was positively correlated with SP-A concentration as well (r(3) = 0.494, P < 0.05). (5) The PC and PI concentrations in amniotic fluid were (65.4 ± 7.2) mg/L and (3.8 ± 0.6) mg/L in ICP group, (69.7 ± 3.7) mg/L and (4.3 ± 0.7) mg/L in control group, respectively. The differences were significant (P < 0.05). The concentration of LPC in amniotic fluid in ICP group was (4.8 ± 0.9) mg/L, significantly higher than that in control group (P < 0.05), which was (4.2 ± 0.6) mg/L. The concentration of SM in amniotic fluid was (3.5 ± 0.8) mg/L in ICP group, (4.0 ± 0.5) mg/L in control group, with no significant difference (P > 0.05). (6)The ratio of PC/LPC in ICP group (14.2 ± 3.2) was significantly lower than that in control group (16.9 ± 2.5) (P < 0.05). (7)The TBA concentration in cord blood was negatively correlated with PC and PI concentrations (r(1) = -0.561, r(2) = -0.407, P < 0.05), and had no correlation with LPC concentration (r(3) = 0.260, P > 0.05). CONCLUSIONS: (1) The fetal TBA concentrations in both cord blood and amniotic fluid of patients with ICP was higher than those of healthy gravidas, they were also positively correlated with maternal TBA concentration. (2) ICP resulted in the change of fetal pulmonary surfactant and this change was associated with TBA concentrations in both cord blood and amniotic fluid.

[Effect of highly active anti-retroviral therapy on prevention of mother to child transmission of HIV and on infant growth and development].

OBJECTIVE: To identify the effect of highly active anti-retroviral therapy (HAART) on prevention of mother to child transmission (PMTCT) of HIV and on infant growth and development. METHODS: A total of 16 HIV-infected women or pregnant women selected in this study received HAART before or 18 - 24 weeks after pregnancy. The treatment included taking Zidovudine (AZT) 0.3 g each time, twice a day, Lamivudine (3TC) 0.3 g each time, once a day and Nevirapine (NVP) 0.2 g each time, twice a day or Efavirenz (EFV) 0.6 g each time, once a day, as well as labor intervention and artificial feeding. The growth index for 17 infants from HIV-infected mothers (experimental group) and 16 normal infants (control group) were observed for 18 months. Neonatal hemoglobin (Hb), liver and kidney function, serum iron and calcium were detected at neonatal period and at 12(th) month, respectively. RESULTS: All the pregnant women were in good conditions and had tolerance with HAART. The birth weight, length and Apgar score of the newborns in the experimental group were (3.5 ± 0.9) kg, (54.2 ± 3.8) cm and 7 - 10 scores respectively, however those in the control group were (3.6 ± 0.8) kg, (55.6 ± 3.6) cm and 8 - 10 scores (t(weight) = 1.01, t(length) = 6.98, P > 0.05). Weight and length of infants in experimental group were (9.36 ± 1.8) kg and (76.3 ± 2.7) cm at 12(th) month, while those in control group were (9.86 ± 2.5) kg and (76.8 ± 2.9) cm (t(weight) = 0.83, t(length) = 1.00, P > 0.05). The level of Hb in experimental group was (126.2 ± 16.7) g/L, and was (148.6 ± 20.5) g/L in control group (t = -5.89, P = 0.11). At 12(th) month, the levels of Hb and the total bilirubin (TB) were (125.9 ± 19.8) g/L and (11.7 ± 3.5) µmol/L in experimental group; and those in the control group were (130.1 ± 18.7) g/L and (13.2 ± 3.7) µmol/L (t(Hb) = -3.82, t(TB) = -2.14, P > 0.05). Serum iron and calcium were (25.4 ± 5.7) µmol/L and (26.4 ± 7.2) µmol/L at neonatal period and were (2.3 ± 0.6) mol/L and (2.8 ± 0.6) mol/L at 12(th) month in experimental group, while those were (26.2 ± 4.9) µmol/L and (28.1 ± 6.9) µmol/L at neonatal period and were (2.6 ± 0.5) mol/L and (3.1 ± 0.5) mol/L at 12(th) month in the control group (t(Fe) = 0.80 and t(Ca) = -3.00 in neonatal period, t(Fe) = -1.50 and t(Ca) = -1.00 at 12(th) month, P > 0.05). All infants of HIV-infected mothers were not infected with HIV when they were 18 months old. CONCLUSION: HAART can prevent mother to child transmission of HIV and it was not found to influence the baby's growth and development in this study.

[Influence of total bile acid in maternal serum and cord blood on neonatal cardiac function from intrahepatic cholestasis of pregnancy].

OBJECTIVE: To study the influence of total bile acid (TBA) of maternal serum and cord blood on neonatal cardiac function in women with intrahepatic cholestasis of pregnancy (ICP). METHODS: The concentration of TBA in the maternal peripheral blood and neonatal cord blood were measured by enzyme in 30 women with ICP as patients group matched with 30 normogravidas as control group. The concentration of cardiac troponin I (cTnI) in cord blood was detected by monoclonal enzyme-linked immunosorbent assay. Fetal left ventricle Tei index was evaluated by color Doppler ultrasonoscope. RESULTS: (1) TBA concentration of maternal serum and cord blood at ICP group were significantly higher than that of control group [(36.0+/-9.6) micromol/L vs. (3.8+/-0.9) micromol/L, (10.1+/-2.0) micromol/L vs. (5.5+/-0.4) micromol/L, P<0.01]. TBA concentration of maternal serum at ICP group was significantly higher than that of neonatal cord blood (P<0.01). Meanwhile, TBA concentration of maternal serum at control group was significantly lower than that of neonatal cord blood (P<0.01). (2) The Tei index of neonatal left ventricle at ICP group were significantly higher than that of control (0.58+/-0.04 vs. 0.43+/-0.03, P<0.01). (3) The concentration of cTnI from cord blood (0.92+/-0.23) microg/L at ICP group were obviously higher than that of control group [(0.52+/-0.10) microg/L, P<0.01]. (4) The TBA concentration of cord blood at ICP group showed positive correlation with maternal blood TBA, cord blood cTnI and fetal left ventricle Tei index respectively (r=0.769, 0.635, 0.758, P<0.01). In the mean time, the positive correlation between the concentration of cTnI from cord blood and fetal left ventricle Tei index was also observed (r=0.637, P<0.01). CONCLUSIONS: Left ventricular dysfunction and myocardial injury were shown in the neonates with ICP, which might be associated with with the elevated TBA level of maternal blood. The Tei index of fetal left ventricle could be used as a reliable parameter to monitor cardiac function and the degree of injured myocardial muscles.

[Relationship of fetal total bile acid and the change of fetal pancreas endocrine secretion and its impact on fetal growth and development in intrahepatic cholestasis of pregnancy].

OBJECTIVE: To investigate the relationship of fetal total bile acid (TBA) concentration with the change of fetal pancreas endocrine secretion and its impact on fetal growth and development in intrahepatic cholestasis of pregnancy (ICP). METHODS: The concentrations of TBA, insulin, glucagon and glucose in the cord blood were measured in 30 fetuses with maternal ICP (case group) and 30 fetuses of normogravidas (control group) after elective cesarean section during the same period in the Department of Obstetrics of Xiangya Second Hospital of Central South University from March 2007 to February 2008. The cord blood TBA concentration was investigated by enzyme method and the concentrations of insulin and glucagon were investigated by radioimmunoassay. The glucose was measured by oxidase-superoxide method. The neonatal weight, length and the ponderal index (PI) were measured after parturition. RESULTS: (1) The cord blood insulin concentration (9.0 +/- 3.3) mU/L and the ratio of insulin over glucagon 0.048 +/- 0.028 in the case group was significantly lower than that of controls (10.1 +/- 3.7) mU/L, 0.050 +/- 0.020 (P < 0.05). The concentrations of TBA (10.3 +/- 3.8) miromol/L and glucagon (235 +/- 57) ng/L in case group were obviously higher than that in controls (4.1 +/- 1.3) micromol/L, (205 +/-34) ng/L (P < 0.05). But no difference was shown in the glucose concentration in cord blood between the case and control groups [(3.4 +/- 1.1) mmol/L vs. (3.6 +/- 1.2) mmol/L, P > 0.05]. (2) The neonatal weight and length in case group were significantly lower than that of control [(3163 +/- 478) g vs. (3498 +/- 393) g, (46.5 +/- 2.3) cm vs. (49.3 +/- 1.9) cm, P < 0.01]; while the Ponderal index in case group was significantly higher than that of control group (3.13 +/- 0.23 vs. 2.92 +/- 0.29, P < 0.01). (3) The cord blood TBA concentration respectively showed a linear relationship with the cord blood insulin concentration, the cord blood glucagon concentration and the ratio of insulin over glucagon in the case group. With the increase in cord blood TBA concentration, the cord insulin concentration and the ratio of insulin over glucagon decreased; meanwhile the cord blood glucagon concentration rose (P < 0.01). The cord blood insulin concentration and the ratio of insulin over glucagon in case group were respectively positively correlated with the neonatal weight and length, and were negatively correlated with the PI (P < 0.01); while the cord glucagon concentration was respectively negatively correlated with the neonatal weight and length, and positively correlated with the PI (P < 0.01). CONCLUSIONS: In ICP fetus pancreas, there are hypoinsulinism, glucagon oversecretion, and decrease of the ratio of insulin over glucagon, which is closely correlated with fetal TBA concentration. The endocrine function of fetal pancreas affects the fetal growth and development.

[Correlation between expression of RhoA and Rho kinase in uterine smooth muscle and the onset of labor].

OBJECTIVE: To investigate the role of RhoA and Rho kinase system in the onset of labor. METHODS: Forty term pregnant women, who delivered through cesarean section at the Xiangya Second Hospital of Central South University from February 2007 to November 2007, were selected and divided into 2 groups: 20 in labor group and 20 in non-labor group. Another 20 non-pregnant women undergoing hysterectomy due to cervical intraepithelial neoplasia were chosen as the control. Real-time fluorescence quantitative RT-PCR and western blot were applied to detect the expression RhoA and ROCK I mRNA and protein in uterine smooth muscle tissue and the correlation between the mRNA and protein expression of RhoA and ROCK I were analyzed. RESULTS: (1) The mRNA expressions of both RhoA and ROCK I were detected in all groups, and higher levels were found in the labor group than in the non-labor group and the control [RhoA mRNA: (3.51 +/- 0.56) x 10(-3) vs. (2.75 +/- 0.52) x 10(-3) and (2.11 +/- 0.54) x 10(-3); ROCK I mRNA: (4.07 +/- 0.66) x 10(-3) vs. (2.71 +/- 0.52) x 10(-3) and (2.01 +/- 0.23) x 10(-3), P < 0.01]. (2) RhoA and ROCK I proteins were also identified in all three groups, and the expressions in the labor and non-labor group were higher than those of the control (RhoA protein: 0.72 +/- 0.23 and 0.64 +/- 0.17 vs. 0.46 +/- 0.15; ROCK I protein: 0.56 +/- 0.14 and 0.42 +/- 0.16 vs. 0.29 +/- 0.08, P < 0.01). (3) The expression of RhoA mRNA and ROCK I mRNA were positively correlated in each of the three groups (r = 0.73, P < 0.01), and the same was found in the expression of RhoA protein and ROCKI protein (r = 0.37, P < 0.01). CONCLUSION: The increased expression of RhoA and Rho kinase may play an important role in the initiation of labor.

[Comparison of five different ultrasonographic parameters for diagnosis of lethal fetal pulmonary hypoplasia].

OBJECTIVE: To evaluate the clinic value of five different ultrasonographic fetal parameters for prenatal diagnosis of pulmonary hypoplasia. METHODS: Two hundred and seventy-one normal singleton pregnancies with well-established dates between 20 and 40 weeks of gestation were studied to establish normal reference range of five different ultrasonographic fetal parameters. The five parameters, which could reflect fetal lung mass, were as follows: lung area/body weight ratio, lung area, thoracic circumference/ abdominal circumference ratio, lung area/thoracic area ratio and lung area/head circumference. Thirty pregnancies with risk factors for pulmonary hyperplasia were studied for the usefulness of five parameters. Two or more standard deviations below the mean control group measurement were considered abnormal. The prenatal ultrasonic diagnoses of pulmonary hyperplasia were confirmed at neonatal follow-up examinations, on autopsy and by pathologic findings. RESULTS: Lung area and lung area/head circumference increased with gestational age, lung area /body weight ratio decreased with gestational age. The relationships among the two ratios (thoracic circumference/abdominal circumference ratio, lung area/thoracic area ratio) and gestational age were relatively constant. Abnormal lung area/body weight ratio had a higher diagnostic accuracy than other parameters. Sensitivity of the parameters, including lung area, lung area/body weight ratio, thoracic circumference/abdominal circumference ratio, lung area/thoracic area ratio and lung area/head circumference were 83%, 97%, 50%, 70% and 87% , respectively. Sensitivity of the lung area/body weight ratio was 95% (20/21 fetuses); specificity, 9/9 fetuses; positive predictive value, 100% (20/20 fetuses); negative predictive value, 9/10; and accuracy 97% (29/30 fetuses). CONCLUSION: Lung area/ body weight ratio is a good predictor of pulmonary hypoplasia.

[Relationship of the occurrence of fetal distress and change of umbilical cord and expression of vasoactive substance in umbilical vein in intrahepatic cholestasis of pregnancy].

OBJECTIVE: To investigate the changes of umbilical cord and the vasoactive substance in umbilical vein in intrahepatic cholestasis of pregnancy. METHODS: By HE staining method we analyzed the pathologic change of umbilical cord of 25 women with intrahepatic cholestasis of pregnancy (ICP) and fetal distress (ICP fetal distress group), 25 ICP women without fetal distress group (ICP control group) and 27 normal pregnancies (control group). The nitric oxide synthase (NOS) and endothelin-1 (ET-1) were detected in human umbilical vein endothelial cells (HUVEC) by immunohistochemistry method. Umbilical vein total bile acid (TBA) and NOS and ET-1 were measured. RESULTS: (1) A remarkable high TBA level was found in umbilical vein in ICP, and it was higher in ICP fetal distress group (19.0 +/- 2.3) micromol/L than in ICP control group (9.0 +/- 1.7) micromol/L (P < 0.05); it was higher in ICP control group than the control group (4.4 +/- 1.5) micromol/L (P < 0.05). (2) A significant difference was found in the endotheliocytes of umbilical vein in ICP fetal distress group compared with ICP control group. The ratio of cells with pathological changes in ICP fetal distress group (92%, 23/25) was higher than ICP control group (68%, 17/25; P < 0.05). The occurrence of the pathological changes was associated with TBA. (3) The expression of eNOS in ICP fetal distress group 0.09 +/- 0.06 was lower than in ICP control group 0.21 +/- 0.08 (P < 0.05), and it was lower in ICP control group than in control group 0.47 +/- 0.07 (P < 0.05). In contrast, the expression of ET-1 in ICP fetal distress group 0.49 +/- 0.08 was higher than in ICP control group 0.32 +/- 0.07 (P < 0.05), and it was higher in ICP control group than control group 0.14 +/- 0.06 (P < 0.05). The expression of inducible nitric oxide synthase (iNOS) in ICP fetal distress group 0.20 +/- 0.04 and ICP control group 0.21 +/- 0.05 was lower than in control group 0.26 +/- 0.04 (P < 0.05), but no significant difference was found in ICP fetal distress group and ICP control group (P > 0.05). (4) The expression of eNOS, iNOS and ET-1 was correlated with umbilical vein TBA in ICP (r1 = -0.88, r2 = -0.45, r3 = 0.9; P < 0.01), respectively. CONCLUSIONS: High level of TBA in ICP is harmful to the umbilical vein endothelium, which is correlated with the raised expression of ET-1, and the decreased expression of eNOS,and iNOS in human umbilical cord endothelium cells. All these changes of umbilical vein may be associated with the occurrence of fetal distress in ICP.

[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.

[Glycosylated hemoglobin test in gestational abnormal glucose metabolism].

OBJECTIVE: To investigate the clinical significance of the determination of glycosylated hemoglobin (HbAlc) in gestational abnormal glucose metabolism. METHODS: The level of fasting plasma glucose (FPG) and HbAlc in 540 normal gravida and 387 pregnant women with abnormal glucose metabolism was determined. Glucose challenge test (GCT) with 50 g glucose was done to those whose level of FPG was normal and 75 g glucose tolerance test (OGTT) was done to those whose GCT was abnormal. The levels of HbAlc of all subjects were assessed. And all subjects were divided into some groups according to the levels of HbA1c, to investigate the relationship between the complication and the levels of HbAlc. RESULTS: The positive rate of HbAlc in abnormal glucose metabolism pregnant women was 20.9%. It was not sensitive if diagnosed only by this sign. Diagnosis by both FPG and this sign could solve this problem. The incidence of complication was increasing with the rise of HbA1c titre in the study group. CONCLUSION: The determination of HbAlc is important in the screening, diagnosing and assessing the prognoses of the gestational abnormal glucose metabolism.

[Bcl-2, TGFbeta1, and apoptosis in placenta tissues in patients with hypertensive disorder complicating pregnancy].

OBJECTIVE: To determine the apoptosis in placenta tissues of patients with hypertensive disorder complicating pregnancy and its relationship with Bcl-2, TGFbeta1, and to explore the etiology of hypertensive disorder complicating pregnancy. METHODS: Forty-five placenta samples were obtained from pregnancies with hypertensive disorder (15 gestational hypertension, 15 mild preeclampsia, and 15 severe preeclampsia) and 45 normal placenta tissues were enrolled from the third-trimester pregnancies. Immunohistochemistry (SP method) was used to study the expression of Bcl-2 and TGFbeta1 in human trophoblasts. Terminal deoxynucleotidyl transferase-dUTP nick end-labeling (TUNEL) was used to quantify the incidence of apoptosis in human trophoblasts. RESULTS: The apoptosis rate and TGFbeta1 expression in hypertensive disorder complicating pregnancy group was higher than that in the control group, but the Bcl-2 expression was significantly lower than the control group (all Ps<0.01). With the aggravation of this illness, the apoptosis rate and TGFbeta1 expression in the gestational hypertension group, mild preeclampsia group, and severe preeclampsia tended to be increasing, but the Bcl-2 expression was decreasing (P<0.001). The apoptosis of placenta villi and TGFbeta1 expression were positively correlated in the severe preeclampsia group and mild preeclampsia group,but the apoptosis of placenta villi and Bcl-2 were negatively correlated (all Ps<0.05). TGFbeta1 and Bcl-2 expressions in the severe preeclampsia group and mild preeclampsia group were negatively correlated (P<0.05). CONCLUSION: Apoptosis of the placental trophoblasts of pregnancies with hypertensive disorder is evidently enhanced. The TGFbeta1 expression increases and the Bcl-2 expression decreases. The imbalance between TGFbeta1 and Bcl-2 expression may induce the hypertensive disorder.

Corticotropin-releasing hormone activates connexin 43 via activator protein-1 transcription factor in human myometrial smooth muscle cells.

Corticotropin-releasing hormone (CRH) and connexin 43 (Cx43) play crucial roles in uterine contraction and the onset of labor. The aim of the present study was to investigate the regulatory effects of CRH on Cx43 expression in human myometrial smooth muscle cells (SMCs) and, potentially, its activation of the c-Fos/activator protein (AP)-1 signaling pathway. Human myometrial SMCs collected from nonpregnant women were treated with different concentrations of CRH. Transient transfection of AP-1 decoy oligodeoxynucleotide (ODN) was used to block AP-1 sites of Cx43. The transcriptional activity of AP-1 was detected by luciferase assay. Cx43 protein expression was visualized by immunofluorescence staining. mRNA and protein expression of c-Fos and Cx43 were demonstrated by real-time quantitative RT-PCR and Western blot, respectively. CRH facilitated Cx43 expression and enhanced AP-1 promoter activity in human uterine SMCs. After CRH treatment, Cx43 expression in the cytoplasm increased significantly. CRH significantly increased mRNA and protein expression of c-Fos and Cx43 in a dose-dependent manner (P < 0.01). A transient transfection of AP-1 decoy ODN did not affect CRH regulation of c-Fos (P > 0.05) but almost completely abolished CRH-induced enhancement of Cx43 expression (P < 0.01). In human primary myometrial SMCs, CRH enhances Cx43 mRNA and protein expression through upregulation of c-Fos expression. Blockade of AP-1 sites to the Cx43 promoter can neutralize the CRH-induced upregulation of Cx43.

[Expression and roles of corticotropin-releasing hormone, cortisol and dehydroepiandrosterone sulfate in preterm labour].

OBJECTIVE: To investigate the effect of corticotropin-releasing hormone (CRH), cortisol and dehydroepiandrosterone sulfate in pathogenesis of preterm labor. METHODS: A cross-sectional study was conducted including 80 pregnant women in the following categories: (1) preterm delivery in labor (PL, n=26), (2) term in labor (TL, n=29), (3) term not in labor (n=25). The expression and localizations of CRH mRNA in placentas and fetal membranes in three groups respectively were examined by in situ hybridization. Radioimmunoassay was used to detect the levels of corticotropin releasing hormone, cortisol and dehydroepiandrosterone sulfate in fetal umbilical cord blood in three groups. RESULTS: (1) By in situ hybridization, we localized CRH mRNA to the syncytiotrophoblasts of placenta, the amniotic epithelial cells and chorion cells. (2) The positive index of expression of CRH mRNA in fetal membranes of PL (5.4 +/- 1.4) and TL, (5.4 +/- 1.5) was higher than that in term not in labor, (2.0 +/- 1.4, P<0.01). And there was no difference between PL and TL (P>0.05). The positive index of expression of CRH mRNA in placentas of PL (5.5 +/- 1.4) and TL (5.4 +/- 1.5) was higher than that in term not in labor (2.7 +/- 1.5, P<0.01). And there was no difference between PL and TL (P>0.05). The expression of CRH mRNA was not distinct between placentas and fetal membranes in three groups (P>0.05). (3) The levels of CRH and DHEA-S in umbilical cord blood of PL (7.8 +/- 3.3) ng/L, and (514 +/- 295) microg/L, respectively and of TL (7.7 +/- 4.1) ng/L, and (483 +/- 207) microg/L, were higher than that in term not in labor (4.8 +/- 2.4) ng/L, and (360 +/- 80) microg/L, respectively (P<0.05). And there was no difference between PL and TL (P>0.05). In PL, the level of CRH in umbilical cord blood and the expression of CRH mRNA in placentas and fetal membranes were correlated with each other (r=0.935 and 0.853, P<0.01). Also in TL, the levels of CRH and CRH mRNA were correlated with each other (r=0.902 and 0.825, P<0.01). (4) The level of cortisol in umbilical cord blood of PL (246 +/- 117) microg/L was higher than that in TL (172 +/- 72) microg/L (P<0.05) and term not in labor (126 +/- 60) microg/L (P<0.01). And the levels of cortisol in umbilical cord blood of TL were higher than that in term not in labor (P<0.05). In PL, the levels of CRH and cortisol and dehydroepiandrosterone sulfate in umbilical cord blood were correlated respectively with each other (r=0.523 and 0.424, P<0.05), and in TL the level of CRH was correlated with cortisol and dehydroepiandrosterone sulfate respectively (r=0.438 and 0.354, P<0.05). CONCLUSIONS: (1) CRH may participate in onset of labor. Increase in CRH may be an important stimulator of preterm labor. (2) Cortisol and dehydroepiandrosterone sulfate play an important role in the initiation of labor.

[Expression and activity change of angiotensin converting enzyme in the placenta of preeclampsia].

OBJECTIVE: To investigate the effect of angiotensin converting enzyme (ACE) in the pathogenesis of preeclampsia. METHODS: A cross-sectional study was conducted with 42 pregnant women in the following categories: 30 cases of preeclampsia (mild preeclampsia, n=15; severe preeclampsia, n=15), and normal pregnancy (control group,n=12). The expression and localization of ACE mRNA in the placenta of the 3 groups were respectively examined by in situ hybridization. Ultraviolet radiation colorimetry was used to detect the activity of ACE in the placenta tissue homogenate and the mothers' serum in the 3 groups. RESULTS: The expression of ACE mRNA was found in the endothelial cells of villus and trophoblasts in the placenta. The positive index of ACE mRNA in the placenta of preeclampsia(3.12+/-0.94) was higher than that in the normal pregnancies(1.65+/-0.67) (P<0.05), and there was significant difference between severe preeclampsia and mild preeclampsia (P<0.05). The levels of ACE activity in the placenta tissue homogenate and the maternal serum of preeclampsia were higher than those in the normal pregnancies (P<0.05), and there was significant difference between severe preeclampsia and mild preeclampsia (P<0.05). The placenta tissue homogenate ACE activity was correlated with ACE activity of the maternal serum (r=0.781,P<0.05). CONCLUSION: The expression and activity of local ACE in the placenta tissue may play an important role in preeclampsia and contribute to the development of preeclampsia.