Phosphoproteomics reveals the apoptotic regulation of aspirin in the placenta of preeclampsia-like mice.

Preeclampsia (PE) is a severe gestational complication, and dysfunctional placenta plays an essential role in PE pathogenesis. Although low-dose aspirin is currently the most promising prophylactic drug for PE prevention, the exact mechanism of aspirin remains unclear. A previous study reported that treatment with low-dose aspirin could ameliorate PE-like symptoms in lipopolysaccharide (LPS)-induced PE-like mouse model. This study aimed to uncover the potential mechanism of aspirin action in PE through quantitative phosphoproteomics comparison. We established the following four groups: a control (CTRL) group, an LPS-treated (L) group, an LPS + aspirin co-treatment (LA) group, and an aspirin-treated (A) group. A total of 4350 phosphosites and 4170 phosphopeptides from 1866 phosphoproteins were identified in the placenta on embryonic day 13.5. Among the significantly altered phosphoproteins identified, apoptosis-related pathways were significantly regulated in both the L group vs. CTRL group and the LA group vs. L group comparisons. We demonstrated that apoptosis was increased in the placenta of PE-like mice and was inhibited in the LA group by quantify the apoptosis-positive cells and the protein levels of cleaved caspase 3, 8, and 9. Moreover, the phosphorylation of HSP90ß (S254) and GSK3ß (Y216) may be a crucial factor in the aspirin-mediated regulation of apoptosis according to protein-protein interaction analysis. This study revealed that apoptosis regulation is a mechanism of aspirin action in PE, particularly in women with over-activated inflammation. The phosphorylation of HSP90ß (S254) and GSK3ß (Y216) could be the key intervention targets.

Lifestyle intervention for gestational diabetes mellitus prevention: A cluster-randomized controlled study.

OBJECTIVE: The study was to examine whether gestational diabetes mellitus (GDM) can be prevented by early trimester lifestyle counseling in a high-risk population. METHODS: From September 2012 to January 2013, 1664 pregnancies in the Department of Obstetrics and Gynecology of First Hospital of Peking University were enrolled in the study during their first prenatal care visit before the 8 gestational weeks visit and asked to fill out a questionnaire on GDM risk evaluation. According to the questionnaire and medical records, those with at least one risk factor of GDM were included in the intervention study and randomly allocated to two groups, intervention group and control group. Routine prenatal care was offered, while standardized two-step lifestyle intervention was provided to the intervention group during 6-8 gestational weeks, and at 12-13 gestational weeks, enforcement intervention based on maternal anthropometrics were offered. Both groups were followed until 75 g oral glucose tolerance test (OGTT) testing at 24-28 gestational weeks. The weight gain after intervention and the prevalence of GDM were used to evaluate the effect. RESULTS: (1) According to the International Association of Diabetes and Pregnancy Study Groups (IADPSG) criteria, the positive rate of GDM for the intervention group was 17.16% (23/134), lower than the control group which was 23.91% (33/138), P = 0.168. (2) The weight gain during the first and second trimester for the intervention group was (1.38 ± 2.34) kg and (5.51 ± 2.18) kg, lower than in the control group which was (1.41 ± 2.58) kg and (5.66 ± 2.25) kg, (P = 0.905, P = 0.567). (3) Positive rate of GDM for those fasting plasma glucose (FPG) ≥5.1 mmol/L during early pregnancy was 11/36 (30.55%) for the intervention group that was lower than 17/37 (45.95%) for the control group, but the statistical difference was not significant (P = 0.076). CONCLUSION: The positive rate of GDM could be reduced by a certain amount lifestyle intervention from the beginning of pregnancy. More validated effective intervention should be explored in the high-risk pregnant women.

Increased hepatic peroxisome proliferator-activated receptor coactivator-1α expression precedes the development of insulin resistance in offspring of rats from severe hyperglycemic mothers.

BACKGROUND: Prenatal hyperglycaemia may increase metabolic syndrome susceptibility of the offspring. An underlying component of the development of these morbidities is hepatic gluconeogenic molecular dysfunction. We hypothesized that maternal hyperglycaemia will influence her offsprings hepatic peroxisome proliferator-activated receptor coactivator-1α (PGC-1α) expression, a key regulator of glucose production in hepatocytes. METHOD: We established maternal hyperglycaemia by streptozotocin injection to induce the maternal hyperglycaemic Wistar rat model. Offspring from the severe hyperglycemia group (SDO) and control group (CO) were monitored until 180 days after birth. Blood pressure, lipid metabolism indicators and insulin resistance (IR) were measured. Hepatic PGC-1α expression was analyzed by reverse transcription polymerase chain reaction and Western blotting. mRNA expression of two key enzymes in gluconeogenesis, glucose-6-phosphatase (G-6-Pase) and phosphoenolpyruvate carboxykinase (PEPCK), were analyzed and compared. RESULTS: In the SDO group, PGC-1α expression at protein and mRNA levels were increased, so were expression of G-6-Pase and PEPCK (P < 0.05). The above effects were seen prior to the onset of IR. CONCLUSION: The hepatic gluconeogenic molecular dysfunction may contribute to the metabolic morbidities experienced by this population.

Role of Toll-like receptor 4 and human defensin 5 in primary endocervical epithelial cells.

BACKGROUND: Endocervical epithelial cells play early roles in the defense of upper female genital tract to pathogens. Toll-like receptors (TLRs) and human defensins (HD) have recently been identified as fundamental components of the innate immune responses to bacterial pathogens. We aimed to use in vitro model of human primary endocervical epithelial cells (HPECs) to investigate their roles in innate immune response of the endocervix. METHODS: TLR4 expression and distribution in HPECs and endocervix were investigated by immunofluorescence (IF). Cultured HPECs were divided into lipopolysaccharide (LPS) group which were treated by LPS for 0, 24 and 48 hours, and control group without treatment. At each time point, the levels of HD5, IL-6 and TNF-alpha in supernants were determined by ELISA. TLR4 and HD5 expressions of cells were detected by Western blotting simultaneously. HD5 expression pattern was also compared between the HeLa cell line and HPECs. RESULTS: Endocervix tissue surface and HPECs expressed TLR4. After incubated with LPS, HPECs expressed significantly higher levels of TLR4 than control group, especially after 24 hours (P < 0.01), however decreased after 48 hours with a similar level of TLR4 expression compared with control group. LPS could upregulate the secretion of HD5, IL-6 and TNF-alpha in a time-dependent manner (24 hours: P < 0.05; 48 hours: P < 0.01, compared with control group). Intracellular HD5 expression levels decreased over time. HD5 expression patterns in HPECs were different from HeLa cell line. CONCLUSIONS: To respond to LPS stimulation, HPECs may function in the mucosal immune defense through TLR4 activation and HD5 secretion. HPEC is considered a significant model for immunological study.

[Preterm birth and preterm infants in Beijing regional district].

OBJECTIVE: To investigate the incidence and relevant information of preterm birth and the outcomes of preterm infants delivered at various gestational weeks and for different causes. METHODS: Totally 955 women, who ended their pregnancies before term, and 1066 neonates of the previous mothers were enrolled in this survey, among 15,197 deliveries at Peking University First Hospital, Beijing Gynecological and Obstetric Hospital, Women's and Children's Hospital of Haidian District and Peking University Third Hospital, respectively, from December 1(st), 2006 to May 31(st), 2007. RESULTS: (1) Incidence of preterm birth: the overall incidence of preterm birth of the 4 hospitals was 6.3% (955/15 197), and it was 8.1% (125/1549) in Peking University First Hospital, 13.1% (150/1142), which was the highest (P < 0.01), in Peking University Third Hospital, 5.5% (369/6656) in Beijing Gynecological and Obstetric Hospital and 34.0% (311/5850) in Women's and Children's Hospital of Haidian District. The preterm birth rate at the two comprehensive hospitals was significantly higher than that of the two specialized hospitals [10.2% (275/2691) vs 5.4% (680/12 506), P < 0.01]. (2) Gestational weeks at delivery: the incidence of preterm birth before 34 weeks was 28.5% (272/954) and the number changed to 71.5% (682/954) for those preterm deliveries after 34 weeks. However, this number varied among the 4 hospitals. Peking University First Hospital had the highest incidence of preterm birth before 34 weeks (P < 0.05), and the lowest was found in Women's and Children's Hospital of Haidian District (P < 0.01), but no difference was found between Peking University Third Hospital and Beijing Gynecological and Obstetric Hospital. (3) Etiology of preterm birth: preterm premature rupture of membranes (PPROM) accounted for the most proportion of all preterm birth cases, followed by iatrogenic preterm birth and spontaneous preterm birth. But the causes of preterm birth in the 4 hospitals were different. Peking University Third Hospital had a higher incidence of iatrogenic preterm birth than the others (P < 0.01), and Peking University First Hospital had a higher incidence of preterm birth caused by PPROM and lower incidence of spontaneous preterm birth. The first four reasons of iatrogenic preterm birth were preeclampsia (143, 42.0%), fetal distress (58, 17.1%), placenta previa (43, 12.6%) and placenta abruption (33, 9.7%). (4) Neonatal outcomes in different hospitals: the neonatal outcomes were quite different among the 4 hospitals due to different causes and different delivery weeks. The highest neonatal mortality rate was found in Beijing Gynecological and Obstetric Hospital (5.4%, 22/408) compared to that in Women's and Children's Hospital of Haidian District (1.3%, 4/320) and Peking University Third Hospital (0.6%, 1/170) (P < 0.01), but without any difference when compared to that in Peking University First Hospital (2.4%, 3/124) (P > 0.05). (5) Neonatal outcomes at different gestational age: the recovery rate of preterm infants delivered at < 32 weeks was lower than those delivered > or = 32 weeks (P < 0.01), and this number rose to 99.6% in those delivered > or = 34 weeks. More infants delivered < 32 weeks were given up for treatment or died during the perinatal period than those delivered > or = 32 weeks, with the neonatal mortality rate of 22.1% for those delivered at < 32 weeks and only 0.3% for those delivered at > or = 34 weeks (P < 0.01). (6) Neonatal outcomes for various causes: the premature neonatal mortality rate for iatrogenic preterm births was higher than that of PPROM (4.9% vs 1.6%, P < 0.05). But the neonatal recovery rates were similar among the PPROM, spontaneous and iatrogenic preterm birth group (P > 0.05). CONCLUSIONS: Preterm birth is associated with high perinatal mortality rate, especially for those delivered before 32 weeks which would be highlighted in prevention. Reduction of the iatrogenic preterm birth, combined with proper prevention of PPROM, is an important issue in decreasing the prevalence of preterm birth.

[Study of local immunity of lower genital tract infections].

OBJECTIVE: To investigate the profile of local immunity of vagina and the immune defense mechanisms against lower genital tract infections. METHODS: Vaginal lavage was collected from healthy women and patients of vulvovaginal candidiasis, bacterial vaginosis, Trichomonol vaginitis, human papilloma virus infection (VVC), and chlamydia trachomatis infection. Each group included 60 cases. The level of interleukin (IL)2, 4, 5, 13, 8 and human defensin 5 (HD5) were detected by enzyme linked immunosorbent assay(ELISA). RESULTS: (1) Cytokine of helper T cell 1(Th1): the level of IL-2 between healthy women and VVC/ bacterial vaginosis (BV)/ trichomonol vaginitis (TV)/ chlamydia trachomatis (CT) patients had no significant difference. The IL-2 level(96 +/- 33) x 10(-3) pg/L of human papilloma virus (HPV) infection patients was significantly higher than that of healthy women (P < 0.05). (2) Cytokine of helper T cell 2 (Th2): the level of IL-4 between healthy women and VVC/CT patients had no significant difference. The level of IL-5 between healthy women and BV patients had no significant difference. The IL-13 level (42 +/- 15) x 10(-3) pg/L of TV patients was significantly higher than that of healthy women (30 +/- 29) x 10(-3) pg/L (P < 0.05). The IL-4 level (103 +/- 28) x 10(-3) pg/L of HPV infection patients was significantly higher than that of healthy women (36 +/- 22) x 10(-3) pg/L (P < 0.05). (3) IL-8: the IL-8 level (5.8 +/- 2.7) pg/L of TV infection patients was significantly higher than that of healthy women (2.6 +/- 2.4) pg/L (P < 0.05). The level of IL-8 between healthy women and BV patients had no significant difference. (4) HD5: the HD5 level of TV, BV, VVC, HPV and CT infection patients were significantly higher than that of healthy women (P < 0.05). CONCLUSIONS: (1) HD5 plays an important role in the defence of vaginal epithelial cell. (2) Th2 may be more important than Thl in lower genital tract infections. (3) IL-8 plays an important role in extrinsic source infections.