[miRNA-148b-3p Influences Glucose Metabolism of Offspring with Maternal Cholestasis by Regulating GLUT1 Expression in Placental Trophoblast Cells].

OBJECTIVE: To investigate the expression of miRNA-148b-3p and its target gene in the placenta between normal pregnant women and pregnant women with intrahepatic cholestasis of pregnancy (ICP) and to explore the possible mechanism of glucose metabolism of offspring with maternal cholestasis. METHODS: There were 30 cases of normal pregnant women and 30 cases of pregnant women with ICP recruited in the study, all of whom underwent cesarean delivery from Mar. 2017 to Jan. 2018. Placenta tissues, maternal blood and cord blood were collected in each case. Maternal blood and cord blood were sent for biochemical detection. miRNA of placenta tissues was extracted and qRT-PCR was used to measure the expression of miR-148b-3p in the placenta. Normal HTR-8 cells were transfected with miR-148b-3p inhibitor/mimics wrapped with lipofectaine3000. qRT-PCR was used to measure the expression of miR-148b-3p, and Western blot was used to measure the expression of glucose transporter 1 (GLUT1) after transfection. RESULTS: Maternal fasting blood glucose (FPG) and its fetal cord blood insulin levels in the ICP group were significantly higher than those of control. The expression of miR-148b-3p in the placenta of ICP group was lower than that of control group ( P<0.05). Compared with inhibitor control group, the expression of miR-148b-3p was decreased in HTR-8 cells transfected with miR-148b-3p inhibitor ( P<0.05), while the expression of GLUT1 was increased ( P<0.05). Compared with mimics control group, the expression of miR-148b-3p was increased in HTR-8 cells transfected with miR-148b-3p mimics ( P<0.05), while the expression of GLUT1 was decreased ( P<0.05). CONCLUSION: miR-148b-3p might participate in glucose metabolism of offspring with maternal cholestasis through the negative regulation of GLUT1 expression in placental trophoblast cells.

[The Application of Internal Iliac Artery Balloon Occlusion in Pernicious Placenta Previa].

OBJECTIVES: To evaluate the clinical application value of internal iliac artery balloon occlusion in pernicious placenta previa. METHODS: We retrospectively reviewed the medical records of the patients of pernicious placenta previa in a single center from Jan, 2010 to Jan, 2015. The patients were divided into two groups, internal iliac artery balloon occlusion group and the control group without endovascular intervention. Blood loss in operation, volume of transfused blood products, caesarean hysterectomy, operating time, hospital days after operation and postoperative morbidity were compared between the two groups. RESULTS: The balloon occlusion group had significantly less blood loss, the volume of transfused blood products, caesarean hysterectomy, hospital day after operation than the control group had. There was no statistical difference in operating time, intensive care units (ICU), hypotension, infection, hypoxemia, bladder injury, bowel obstruction, neonatal asphyxia between the two groups. The balloon occlusion group had significantly higher rate in coagulopathy, hypoalbuminemia, electrolyte imbalance. Among the patients whose uterus were preserved, the blood loss was not significantly difference between the two groups. Among the patients with the complication of placenta accreta, caesarean hysterectomy was less in balloon group, and blood loss between the two groups was not significantly different. Among the patients without placenta accrete, the blood loss was less in balloon group, and caesarean hysterectomy between the two groups was not significantly different. The risk of hysterectomy in balloon group was related to placenta accreta, uterine arteries engorgement, placental invasive serosa, taking placenta by hand, placental invasive bladder, barrel-shaped thickening of lower uterine segment, unable to remove placenta. CONCLUSIONS: Internal iliac artery balloon occlusion is an effective treatment for pernicious placenta previa.

[Perinatal Management for Monochorionic Twin Reversed Arterial Perfusion Sequence].

OBJECTIVE: To evaluate the perinatal managementof monochorionic twin pregnancies complicated by twin reversed arterial perfusion (TRAP) sequence. METHODS: A retrospectively study was performed on the management and perinatal outcome of monochorionic multiple pregnanciescomplicated by TRAP sequence at West China Second University Hospital from May 2010 to May 2014. RESULTS: Thirteen cases of TRAP sequence were identified during the study period, included 4 monochorionicmonoamniotic (MCMA) twins, 7 monochorionic diamniotic(MCDA) twins,1 monochorionic-triamniotic (MCTA) triplet pregnancy and 1 MCDA triplet pregnancy. Gestational age at diagnosis of TRAP sequence was from 11+5 to 31+6 gestational weeks in 12 cases. TRAP sequence was diagnosed by post-mortem examination in the case of MCDA triplet pregnancy transferred to our hospital with inevitable abortion at 21+3 weeks. 9 cases underwent conservative management. In the conservative management group, intrauterine death of the pump twin occurred in two MCMA twins and 7 cases delivered a healthy pump twin between 31+3 and 39+5 weeks of gestation. 2 cases were treated with bipolar cord coagulation of acardiac twin and delivered a healthy pump twin at 32+1 and 33+5 weeks of gestation. CONCLUSION: Early antenatal diagnosis of TRAP sequence is very important. Consultation with the parents is recommended as to the options of conservative management or intervention. Conservative management with close monitoring may be a safe option for TRAP sequence with a small acardiac twin. Bipolar cord coagulation of acardiac twin is a relatively safe and effective procedure in TRAP sequence with indications to intervention.

[Perinatal management of twins with discordant congenital defects].

OBJECTIVE: To review the outcomes of perinatal management of twins with discordant congenital defects. METHODS: We retrospectively examined the cases of twins with discordant congenital defects treated in the West China Second University Hospital from December 2011 to December 2013. RESULTS: There were 26 cases of twins (14 dichorionic and 12 monochorionic) with one anomalous fetus. Of those twins, 16 were conceived by nature and 10 by in vitro fertilization and embryo tansfer (IVF-ET). Counselling services were offered to the parents by a multidisciplinary team about options of pregnancy. Termination of pregnancy was chosen on three monochorionic twins. Twelve pairs of twin were delivered at 26(+3)-37(+6) weeks gestation. One pair ended with neonatal death, and another one with gastroschisis was given intrapartum fetal operation. Selective termination was chosen on 11 cases using intracardiac injection of potassium chloride under ultrasonographic guidance (9 cases) or bipolar cord coagulation (2 cases). This resulted in ten live births delivered at 25(+5)-38(+4) gustation and one neonatal death. CONCLUSION: Early diagnosis of twins with discordant congenital defects is important. Multidisciplinary counselling services to parents are recommended for determination of options. Intensive prenatal care is essential in management of twins with discordant congenital defects.

REDD1 expression in placenta during human gestation.

The aim of the study was to detect the regulated in development and DNA responses (REDD1) in human placentas throughout different gestational ages (GAs) and to correlate REDD1 with preeclampsia (PE). In experiments, REDD1 messenger RNA and protein expression levels throughout the gestation were determined using immunohistochemistry, quantitative reverse transcriptase polymerase chain reaction, and Western blot. Furthermore, REDD1 protein levels in placenta were also compared between normal outcome pregnancies (n = 20, GA >37 weeks) and PE pregnancies (n = 29), which includes early onset PE; n = 15, GA: 24-33 weeks) and late onset PE (n = 14, GA: 34-39 weeks) by Western blot. As a result, REDD1 protein was predominantly observed in the cytoplasm of trophoblasts. Moreover, higher levels of REDD1 were found not only at earlier gestational stage but also in the PE groups (P < .05). In conclusion, REDD1 may play an important role in maintaining the normal function of placenta during various stages of gestation and predicted that the increase in REDD1 is related to the pathogenesis of PE.

[Sequential preparation of microvlllous and basal membranes from human placenta].

OBJECTIVE: To improve the technology of isolating paired fractions of the maternal-facing membranes (MVM) and fetal-facing plasma membranes (BM) from a term placenta. METHODS: The component of buffer was improved based on Illsley method. The time of Mg2+ -aggregated basal membranes was extended. MVM were obtained from the supernatant of low speed centrifugation while BM were further purified on a sucrose step gradient. RESULTS: Yield for MVM and BM prepared by the method were (0.55 +/- 10.10) mg/g and (0.54 +/- 0.02) mg/g wet weight of placenta. They were enriched 16.87-fold and 11.19-fold as determined by the membrane marker enzymes, alkaline phosphatase (MVM) and adenylate cyclase (BM). CONCLUSION: The modified Illsley method can easily produce both MVM and BM of satisfied quantity from human placenta. It could be applied as a cell molecular model of maternal-fetal exchange interface.

[Study on relationship between expression of familial intrahepatic cholestasis 1 mRNA in placenta and Intrahepatic cholestasis of pregnancy].

OBJECTIVE: By detecting the expression of placental familial intrahepatic cholestasis 1 (FIC1) and levels of total bile acids (TBA) and cholylglycine (CG) in maternal and umbilical cord serum, the effect of placental bile salt transporter on fetal pathology of intrahepatic cholestasis of pregnancy (ICP) is explored. METHODS: TBA and CG levels in maternal and umbilical cord serum of 20 gravidas complicated with ICP (ICP group) and 20 normal gravidas (control group) of late pregnancy were measured by velocimetry and radioimmunoassay respectively. The placental FIC1 mRNA expression was tested by real time RT-PCR. Meanwhile FIC1 mRNA expression of 4 random placentas were localized by in-situ hybridization. RESULTS (1) TBA and CG levels in both maternal and umbilical cord serum of ICP group were significantly higher than those of control group (P < 0.05). TBA and CG levels in maternal serum were significantly higher than those in umbilical cord serum of ICP group (25.77 +/- 16.64) micromol/L vs (8.55 +/- 5.48) micromol/L for TBA, and (3416.09 +/- 1986.04) microg/dL vs (821.84 +/- 673.17) microg/dL for CG, P < 0.05). However, there were no significant differences between TBA and CG levels in maternal serum and umbilical cord serum from control group (3.4 +/- 2.51) micromol/L vs (4. 36 +/- 3. 26) micromol/L for TBA, and (342.74 +/- 234.88) microg/dL vs (309.32 +/- 145.20) pg/dL for CG, P > 0.05). (2) FIC1 mRNA was expressed and localized to syncytiotrophoblast in both ICP and control placentas. Placental FIC1 mRNA expression was significantly decreased in ICP group than in control group (P < 0.05). (3) There were no significant correlations between TBA and CG levels of maternal or umbilical cord serum and expression of placental FIC1 mRNA in both groups (r = -0.229-0.163, P > 0.05). CONCLUSION: Our research results suggest that the decreased expression of FIC1 mRNA in placenta may be one of actor of disturbing placental bile salt transport and resulting in fetal cholestasis in ICP.

[The expression of human leukocyte antigen G and E on human first trimester placenta and its relationship with recurrent spontaneous abortion].

OBJECTIVE: To investigate the distribution of human leukocyte antigen (HLA)-G and E on human first trimester placenta and its relationship with unexplained recurrent spontaneous abortion (RSA). METHODS: Fifteen women with normal first trimester pregnancy and fifteen patients with RSA were included in this study. In situ hybridization and immunohistochemical staining were employed to detect the mRNA and protein of HLA-G, E in first trimester placenta. RESULTS: The HLA-G, E mRNA were detected in all trophoblasts, including syncytiotrophoblast, villous and extravillous cytotrophoblast. HLA-G protein was only found at extravillous trophoblast by mAb 4H84 specific to HLA-G. The same expression sites were observed for both HLA- E protein and its mRNA. The expression of HLA-G, E mRNA in the RSA group were much lower than those in the control group. The HLA-G, E protein were also expressed at low level in RSA group. CONCLUSION: The inconsistency of the distribution of HLA-G mRNA and protein was probably due to the monoclone antibody 4H84. There is strong correlation between lack of the mRNA and protein expression of HLA-G, E found in the trophoblasts at the fetal-maternal interface and RSA.

[Study of fetal lymphocyte of intrahepatic cholestasis of pregnancy].

OBJECTIVE: To explore effect of fetal lymphocyte on pathogenesis of intrahepatic cholestasis of pregnancy (ICP). METHODS: Twenty pregnant women with ICP and 20 normal pregnant women were enrolled in the study. The single mixed lymphocyte culture/reaction (MLC/MLR) was conducted using inactive lymphocyte obtained from maternal peripheral blood and lymphocyte of cord blood from fetus. Antigen-induced-lymphocyte-proliferation-reaction was used for dermic soluble antigen and decidual soluble antigen obtained from maternal blood and cord blood from fetus. The intense of proliferation was calculated and compared between normal and ICP-complicated pregnancies. RESULTS: (1) The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 2.75 +/- 0.36 than those of normal control group 1.45 +/- 0.19 in single mixed lymphocyte culture (P < 0.05). (2) The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 1.45 +/- 0.19 than those of normal control group 0.67 +/- 0.24 in decidual soluble antigen induced lymphocyte proliferation reaction (P < 0.05). (3) The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group (1.22 +/- 0.44) than those of normal control group (0.66 +/- 0.27) in dermic soluble antigen induced lymphocyte proliferation reaction. CONCLUSIONS: (1) The fetal lymphocyte may be one of the effector cells in pathogenesis of ICP. (2) The disturbance of fatal-maternal immune-tolerance is one of the important mechanisms underlying ICP.

[Expression and localization of estrogen receptor ERalpha and the kinds of ERbeta mRNA on placenta].

OBJECTIVE: To investigate the kinds of ERa, ERbeta mRNA and the localization of ERa, ERbeta on normal human placenta. METHODS: The expression and localization of ERa,ERbeta on normal human placenta were detected by reverse transcription-ploymerase chain reaction (RT-PCR) and immunhistochemistry (SP-assay). RESULTS: There was mRNA expression of ERalpha and ERbeta on human term placenta, and ERbeta was identified with ERbeta5. The ERa immunoreactivity on human term placenta was in villous cytotrophoblastic nucleus, and that of ERbeta was in syncytiotrophoblastic cytoplasm. CONCLUSION: The findings indicate that ERbeta5 is the kind of ERbeta on placenta. It is localizated in the cytoplasm of syncytiotrophoblast and is probably connected with the production of estrogen.

[VCAM-1 and E-selectin expression in extravillous cytotrophoblasts of severe preeclampsia].

OBJECTIVE: To investigate the expression pattern of vascular cell adhesion molecule-1(VCAM-1) and endothelial selectin (E-selectin) in the placental bed extravillous cytotrophoblast (EVCT) as well as its relationship with preeclampsia. METHODS: Placentas from severe preeclampsia (n=20) and gestional age-matched normal pregnancies (n=20) were studied. The staining intensity and the Mean Optical Density (MOD) of VCAM-1 and E-selectin in EVCT of placental bed deciduas were measured by immunohistochemistry. RESULTS: The staining intensity and MOD of VCAM-1 and E-selectin on placental bed EVCT in severe preeclampsia were significantly lower than those in the control group respectively (P<0.05, P<0.01). CONCLUSION: The decreased EVCT expression of vascular adhesion phenotype VCAM-1 and E-selectin may account for the shallow trophoblast invasion in preeclampsia, which is the major pathological change of this disease.

[Study on the association between estrogen receptor alpha gene polymorphism and intrahepatic cholestasis of pregnancy].

OBJECTIVE: To investigate the relationship between estrogen receptor alpha (ERalpha) gene polymorphism and intrahepatic cholestasis of pregnancy (ICP). METHODS: The Xba I and Pvu II polymorphisms in intron 1 of ER alpha gene were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 100 pregnant women with ICP (ICP group) and 100 normal pregnant women (control group). RESULTS: (1) The frequencies of XX, Xx and xx genotypes of Xba I polymorphism were 6%, 33% and 61% respectively in control group, and were 2%, 33% and 65% respectively in ICP group. There was no significant difference in the distribution of Xba I genotypes between the two groups (P > 0.05). The frequencies of the two alleles X and x were 23% and 78% in control group, and were 19% and 82% in ICP group, respectively. There was also no significant difference in the frequency of Xba I alleles between the two groups (P > 0.05). (2) The frequencies of PP, Pp and pp genotypes of Pvu II polymorphism were 18%, 42% and 40% respectively in control group, and were 12%, 53% and 35% respectively in ICP group. There was no significant difference in the distribution of Pvu II genotypes between the two groups (P > 0.05). The frequencies of the two alleles P and p were 39% and 61% in control group, and were 39% and 62% in ICP group, respectively. There was also no significant difference in the frequency of Pvu II alleles between the two groups (P > 0.05). (3) There was also no significant difference in the distribution of Xba I and Pvu II combinative genotype between the two groups (P > 0.05). CONCLUSION: The ERalpha gene polymorphism is not associated with the risk of ICP.

[Perinatal outcomes of intrahepatic cholestasis of pregnancy: analysis of 1210 cases].

OBJECTIVE: To study the perinatal outcomes of intrahepatic cholestasis of pregnancy (ICP). METHODS: The clinical data of 1210 cases of ICP in recent ten years were retrospectively analyzed. RESULTS: The incidence rates of perinatal outcomes of ICP were as follows: 19.0% (230/1210) for threatened premature labor, 24.0% (290/1210) for premature delivery; 23.2% (281/1210) for meconium stained amniotic fluid, 7.1% (86/1210) for neonatal asphyxia, 22.5 per thousand (27/1210) for perinatal mortality, 85.9% (1039/1210) for cesarean section, 0.9% (11/1210) for fetal growth restriction (FGR), 1.4% (17/1210) for postpartum hemorrhage, and 8.1% (101/1210) for preeclampsia. Threatened premature labor occurred beyond the gestation gestation period of 32 weeks in 88.7% (204/230) of the patients, and the fetal death rate in threatened premature labor was 46.7% (7/15). Premature delivery occurred after 34 weeks of gestation in 96.2% of the patients (279/290) 89.7% (260/290) of which were caused by cesarean section because of abnormal fetal monitoring. 41.3% of the cases with meconium stained amniotic fluid (116/281) occurred before the onset of labor. Fetal death accounted for 56% (15/27) of perinatal death, 80% (12/15) of which happened after the gestation week of 35 (36.5 +/- 1.2) with normal fetal heart rate monitoring. 95% (19/20) of the fetal death and stillbirth occurred after threatened premature labor and occasional uterine contractions, or at the early stage of labor. CONCLUSION: The rates of FGR, postpartum hemorrhage, and preeclampsia in ICP are almost the same as those of the normal pregnancy. Routine fetal heart rate monitoring methods cannot predict fetal death. The important measures to decrease the perinatal mortality include paying attention to fetal monitoring when threatened premature labor, occasional uterine contractions and prenatal meconium occur, and at the early stage of labor, and management of threatened premature labor and timely intervention of pregnancy (at the gestation period of 34 - 37 weeks).

[Study of estradiol on treatment of preeclampsia in rat model].

OBJECTIVE: To evaluate whether estradiol can inhibit and cure the inflammation of experimental preeclampsia in rats. METHODS: Experimental preeclampsia was induced in 14-day-pregnant rats by infusion of endotoxin (1.0 microg/kg). Rats with normal pregnancy were infused with sodium chloride solution. A group of preeclampsia rats was injected with 17beta-estradiol (17beta-E(2), 1 mg.kg(-1).d(-1)). Blood pressure, albuminuria, inflammation associated adhesion molecule CD(49d) and tumor necrosis factor-alpha (TNF-alpha) were assessed. RESULTS: On pregnant day 19, for normal pregnancy group (group C) the blood pressure was (120.4 +/- 2.0) mm Hg (1 mm Hg = 0.133 kPa), urinary protein (0.47 +/- 0.06) mg/24 hours; for experimental preeclampsia group (group A) blood pressure was (134.2 +/- 2.4) mm Hg, urinary protein (0.79 +/- 0.10) mg/24 hours; for experimental preeclampsia with 17beta-E(2) treatment group (group B) blood pressure was (123.3 +/- 1.7) mm Hg, urinary protein (0.51 +/- 0.08) mg/24 hours. A significant increase of blood pressure and urinary albumin was observed in group A. CD(49d) expression and TNF-alpha concentration were also increased. 17beta-E(2) reduced the expression of CD(49d), concentration of TNF-alpha, blood pressure and albuminuria of experimental preeclampsia. However, the weight of fetuses in 17beta-E(2) treatment group were less than that in other groups. CONCLUSION: 17beta-E(2) can improve the symptoms of experimental preeclampsia, but its effects on fetus need to be further studied.