Pigment epithelium-derived factor/vascular endothelial growth factor ratio for early prediction of preeclampsia: A prospective multicenter study in China.

BACKGROUND: Imbalance of circulating factors related to angiogenesis plays a central role in the pathogenesis of preeclampsia (PE). OBJECTIVE: The aim of this study was to discover and validate a cutoff value of pigment epithelium-derived factor (PEDF)/vascular endothelial growth factor (VEGF) ratio for early prediction of PE before 20 weeks of gestation. STUDY DESIGN: This prospective multicenter study was performed in mainland China, and was divided into 3 phases to discover, develop, and validate a cutoff value of PEDF/VEGF ratio that could predict PE prior to diagnosis in pregnant women at high risk of PE (12 weeks 0 days to 19 weeks 6 days of gestation). We estimated PEDF/VEGF ratio at 5 visits: from visit 0 (baseline) to the postpartum visit. RESULTS: In the discovery phase (200 women), we found that antiangiogenic PEDF was higher and angiogenic VEGF was lower in the PE group than in the control group before 20 weeks of gestation. In the development phase (650 women), we found that a cutoff value of 800 for PEDF/VEGF ratio demonstrated a preferably predictive value. Subsequently, in the validation phase (additional 900 women), we found that the negative predictive value of PEDF/VEGF ratio ≤800 at the visit 1 was 98.6% (95% CI, 97.3-99.4), at the visit 2 was 96.9% (95% CI, 95.1-98.1) and at the visit 3 was 95.1% (95% CI, 93.0-96.7). ORs were 4.40, 6.27, and 5.73, respectively. CONCLUSIONS: PEDF/VEGF ratio ≤800 may have some predictive value for early diagnosis of PE. Further multicenter studies with larger sample sizes are necessary to confirm our findings.

Plasma Level of Placenta-Derived Macrophage-Stimulating Protein -Chain in Preeclampsia before 20 Weeks of Pregnancy.

OBJECT: This study aimed to investigate the diagnostic value of placenta-derived macrophage-stimulating protein α-chain (MSP-α) before the 20th week of gestation for the early diagnosis of preeclampsia (PE). METHODS AND MATERIALS: Two parts of this nested case-control study were simultaneously executed, and 1500 pregnant women were recruited. A total of 124 pregnant women were included in the plasma analysis part of this study. The MSP-α plasma level was measured before the 20th week of gestation, and the participants were followed until delivery. A case group of 62 women with PE and a control group of 62 women matched by gestational age, maternal age, and pre-pregnancy BMI (with normotensive pregnancies) were evaluated. In the placenta analysis part of this nested case-control study, the placentas of 34 pregnant women were randomly obtained. The placental levels of MSP were measured in 17 individuals with PE (case group) and in 17 women with a normotensive pregnancy matched by gestational age and maternal age (control group). RESULTS: The plasma level of MSP-α was higher in the PE group than in the control group before the 20th week of gestation (p < 0.001). In addition, compared to the women with severe features in the PE group, those without severe features had a significantly higher plasma MSP-α level before the 20th week of gestation (p < 0.001). The area under the receiver operating characteristic curve (AUC) of MSP-α before the 20th week of gestation was 0.905 (95% CI, 0.811-0.962) for the women with early-onset PE without severe features. With regard to the placenta, the PE group (accumulated optical density, IOD [SUM] = 8862.37 ± 2064.42) exhibited increased MSP staining (more intense MSP staining or more extensive staining) compared with the control group (normal pregnancies (IOD [SUM] = 447.92 ± 114.72, P < 0.001). Furthermore, increased MSP staining was detected among the women without severe features compared with those with severe features in the PE group (IOD [SUM]: 12192.65 ± 5325.56 vs. 4104.83 ± 2383.06, P = 0.021). CONCLUSION: According to the findings of this study, the plasma level of MSP-α may be associated with PE, and MSP-α may be considered a candidate protein for further analysis in studies of PE. Multicenter studies with larger sample sizes must be performed in the future to obtain accurate results regarding the predictive value of MSP-α in combination with other protein factors for the early diagnosis of PE.

TRAIL Is Decreased Before 20 Weeks Gestation in Women with Hypertensive Disorders of Pregnancy.

OBJECTIVE: The present study evaluated maternal plasma protein profiles before the onset of hypertensive disorders of pregnancy (HDP) to assess the relationship between maternal plasma tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and HDP before 20 weeks gestation and to evaluate the discriminatory performance of plasma TRAIL levels for HDP. METHODS: A 2-phase discovery/validation study was designed. In the discovery phase, a nested case-controlled study was performed using plasma sampled at 8 to 20 weeks gestation from 20 women who later developed HDP and from 20 age- and gestational week-matched controls. Plasma was analyzed using a human protein microarray technology designed to simultaneously detect 507 proteins. The functional annotation and clustering of the differentially expressed proteins were performed using DAVID and the GO database. TRAIL levels were further validated in an independent study using plasma obtained at 8 to 20 weeks gestation from 53 women who later developed HDP and from 106 matched controls, and 62 clinical risk factors were investigated. RESULTS: In the protein microarray analysis, 23 proteins were differentially expressed between the two groups. The ELISA showed that women who later developed HDP had significantly lower TRAIL levels compared to women with uncomplicated pregnancies. The multivariable Cox regression analysis identified the following three factors that were entered into the final Cox regression model: gravidity (OR = 2.02, 95% CI 1.00-4.09), pre-pregnancy BMI (OR = 1.46, 95% CI 1.21-1.76) and TRAIL levels (OR = 0.97, 95% CI 0.94-0.99). The model had a significantly better discriminatory power (AUC = 0.83, 95% CI 0.75-0.88) compared to TRAIL alone as an independent predictor of HDP (AUC = 0.59, 95% CI 0.51-0.67). CONCLUSION: Twenty-three differentially expressed proteins before 20 weeks gestation might be associated with the pathogenesis of HDP. Plasma TRAIL levels were associated with the development of HDP, and the combination of plasma TRAIL levels with pre-pregnancy BMI and gravidity had a good discriminatory performance for HDP before 20 weeks gestation.

[Establishment and application of co-transfection screening method for phytoestrogen active constituents].

OBJECTIVE: To establish a highly sensitive screening method for phytoestrogen active constituents and to primarily screen the phytoestrogenic active constituents from the chickpea extractions by the method. METHOD: Human ERalpha cDNA was cloned using MCF-7 total RNA as the template by RT-PCR and then was constructed into a pcDNA3 and named as pERalpha. The cell line MCF-7 was co-transfected with pERalpha and the reporter plasmid pERE-Luc which carrying the estrogen response element (ERE) plus the luciferase reporter gene. The luciferase activity was then assayed. The model was optimized by changing the ratio of two plasmids. The feasibility of the optimized model was further proved by the several known phytoestrogen compounds including fermononetin, biochanin A and genistein, et al. As an application of the model, the phytoestrogen activity of the extracts of the chickpea was assayed. RESULT: The recombinant plasmid (pERalpha) can enhance luciferase activities of pERE-Luc transfected MCF-7 cells. The highest transfection efficiency and luciferase activity were found at the ratio of 10:1 (pERE-Luc: pERalpha), the luciferase activity was improved five times as high as the unique pERE-Luc transfection. The co-transfection screening model also indicated that fermononetin, biochanin A and genistein could induce ERE-driven luciferase activity and ICI 182,780 suppressed the induced transcription. As the application of the model, the results showed that the ethanol (70%) total extraction, the ethyl acetate extraction and the ligarine extraction of the chickpea can induce ERE-driven luciferase activity. Concurrent treatment with ICI 182,780 abolished the induced luciferase activity. CONCLUSION: A phytoestrogen active constituent screening mode have been established based on co-transfection method. It is sensitive to assay the phytoestrogen active constituents and can be applied to screen the active component of phytoestrogens.