Effects of Notch2 and Notch3 on Cell Proliferation and Apoptosis of Trophoblast Cell Lines.

AIMS: To investigate the effect of Notch2 and Notch3 on cell proliferation and apoptosis of two trophoblast cell lines, BeWo and JAR. METHODS: Notch2 and Notch3 expression in BeWo and JAR cells was upregulated or downregulated using lentivirus-mediated overexpression or RNA interference. The effect of Notch2 and Notch3 on cell proliferation was assessed by the CCK-8 assay. The effect of Notch2 and Notch3 on the apoptosis of BeWo and JAR cells was evaluated by flow cytometry using the Annexin V-PE Apoptosis kit. Lentivirus-based overexpression vectors were constructed by cloning the full-length coding sequences of human Notch2 and Notch3 C-terminally tagged with GFP or GFP alone (control) into a lentivirus-based expression vector. Lentivirus-based gene silencing vectors were prepared by cloning small interfering sequences targeting human Notch2 and Notch3 and scrambled control RNA sequence into a lentivirus-based gene knockdown vector. The effect of Notch2 and Notch3 on cell proliferation was assessed by the CCK-8 assay. And the effect of Notch2 and Notch3 on the apoptosis of BeWo and JAR cells was evaluated by flow cytometry using the Annexin V PE Apoptosis kit. RESULTS: We found that the downregulation of Notch2 and Notch3 gene expression in BeWo and JAR cells resulted in an increase in cell proliferation, while upregulation of Notch3 and Notch2 expression led to a decrease in cell proliferation. Moreover, the overexpression of Notch3 and Notch2 in BeWo and JAR cells reduced apoptosis in these trophoblast cell lines, whereas apoptosis was increased in the cells in which the expression of Notch3 and Notch2 was downregulated. CONCLUSIONS: Notch2 and Notch3 inhibited both cell proliferation and cell apoptosis in BeWo and JAR trophoblast cell lines.

Expression of notch family proteins in placentas from patients with early-onset severe preeclampsia.

OBJECTIVES: This study is aimed to identify the expression of Notch family proteins in placentas from patients with early-onset severe preeclampsia. STUDY DESIGN: The expression of Notch family proteins in placentas was investigated by immunohistochemistry, Western blotting, and real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The profile of distribution of all Notch family proteins in placentas from patients with early-onset severe preeclampsia is similar to that in normal placentas. All Notch family proteins are expressed in placental trophoblasts. Moreover, Notch1 and Jagged1 (Jag1) are detected in placental endothelial cells. Real-time RT-PCR showed that messenger RNA levels of Notch2 and Delta-like4 (Dll4) in placentas from patients with early-onset severe preeclampsia are lower than that of normal placentas. Western blotting showed a significant increase in Notch3 expression and a significant decrease in Notch2 expression in placentas from patients with early-onset severe preeclampsia relative to those in normal placentas. CONCLUSION: The results suggest that Notch2 and Notch3 may play some roles in the pathogenesis of preeclampsia.

[Analysis of clinical features of pregnant women with hypertrophic cardiomyopathy].

OBJECTIVE: To discuss the clinical features of pregnant women with hypertrophic cardiomyopathy (HCM). METHODS: There were 28 patients with HCM who delivered in Renji hospital of Shanghai Jiaotong University from January 2000 to August 2012. Clinical data were analyzed, including diagnosis, cardiac functional grading, gestational weeks of delivery, delivery mode, birth weight, Apgar scores, etc. RESULTS: (1) Of all the 28 patients, 14 (50%) were diagnosed before pregnancy and others (50%) were diagnosed during pregnancy.(2) Four cases were obstructive HCM (14%), 3 with cardiac function grade I and 1 with grade II. Twenty four cases were non-obstructive HCM (86%), 14 with cardiac function grade I, 9 with grade II and 1 with grade IV. (3) Of all the 28 patients, 4 had family history, 18 (64%) had clinical symptoms or signs which occurred in 8-32 gestational weeks. Twenty-three cases had abnormal ECG (82%). Among them 21 had non-obstructive HCM (88%), with average interventricular septal thickness of (22 ± 3) mm. The other 2 patients had obstructive HCM, with average interventricular septal thickness of (23 ± 4) mm.7 patients (7/28, 25%) had mild-to-moderate pulmonary hypertension [6 with non-obstructive HCM (6/24, 25%) ], and 10 patients had abnormal myocardial enzyme spectrum or troponin levels [9 with non-obstructive HCM (9/24, 38%) ].(4) Among all the patients, only one had vaginal delivery and others received cesarean section. Twenty-two patients had term pregnancies and 6 had preterm birth. The average gestational weeks of delivery in non-obstructive HCM and obstructive HCM were (36.5 ± 2.5) and (38.5 ± 0.4) weeks, respectively. The average birth weight of neonates were (2684 ± 563) and (3164 ± 321) g, and Apgar scores were 9.9 and 10 (10 minutes) respectively. Patients transferred to NICU after delivery were 8 and 0. There was 1 maternal death (with non-obstructive HCM whose ejection fraction was only 26%) and no perinatal death. CONCLUSIONS: More attention should be paid to the clinical signs and abnormal ECG.HCM could be definitely diagnosed by timely echocardiography. Patients with hypertrophic cardiomyopathy were mainly non-obstructive HCM, with cardiac function grade I and II. Monitoring the change of ejection fraction during pregnancy would help. Perinatal outcomes were fine.

Risk factors and risk index of cardiac events in pregnant women with heart disease.

BACKGROUND: Pregnant women with heart disease are at high risk. Studies of risk factors of these patients are of great significance to improve maternal and fetal outcomes. In this paper, we try to discuss the main risk factors of cardiac events in pregnant women with heart disease and to establish a risk assessment system. METHODS: A retrospective analysis was carried out for pregnancies in 1741 women with heart disease who delivered in Shanghai Obstetrical Cardiology Intensive Care Center between January 1993 and September 2010. A Logistic regression model was used to identify independent risk factors of cardiac events and calculate the risk index in pregnant women with heart disease. RESULTS: The composition of heart disease in pregnant women was arrhythmia (n = 662, 38.00%), congenital heart disease (CHD; n = 529, 30.40%), cardiomyopathy (n = 327, 18.80%), rheumatic heart disease (RHD; n = 151, 8.70%), and cardiopathy induced by pre-eclampsia (n = 53, 3.00%). Main cardiac events were heart failure (n = 110, 6.32%), symptomatic arrhythmia needing medication (n = 43, 2.47%), cardiac arrest (n = 2, 0.11%), syncope (n = 3, 0.17%), and maternal death (n = 10, 0.57%). Six independent risk factors to predict cardiac events in pregnant women with heart disease were cardiac events before pregnancy (heart failure, severe arrhythmia, cardiac shock, etc., P = 0.000), New York Heart Association (NYHA) class > II (P = 0.000), oxygen saturation < 90% (P = 0.018), pulmonary artery hypertention (PAH) > 50 mmHg (P = 0.025), cyanotic heart disease without surgical correction (P = 0.015), and reduced left ventricular systolic function (ejection fraction < 40%, P = 0.003). Every risk factor was calculated as 1 score. The incidence of cardiac events in patients with scores 0, 1, 2, 3, and ≥ 4 was 2.10%, 31.61%, 61.25%, 68.97%, and 100.00% respectively. CONCLUSIONS: Pregnancy with heart disease could lead to undesirable pregnancy outcomes. The risk of cardiac events in pregnant women with heart disease could be assessed by risk index.