Angiogenic growth factor levels in maternal and fetal blood: correlation with Doppler ultrasound parameters in pregnancies complicated by pre-eclampsia and intrauterine growth restriction.

OBJECTIVES: To correlate levels of angiogenic growth factors with Doppler ultrasound parameters in pregnancies complicated by pre-eclampsia and intrauterine growth restriction (IUGR). METHODS: In 16 women with pre-eclampsia and 15 women with isolated IUGR, pulsatility indices (PI) in the umbilical and uterine arteries were measured by Doppler ultrasonography. At delivery, maternal and fetal blood (umbilical vein and artery separately) was sampled and angiogenic growth factors measured by means of enzyme linked immunosorbent assay (ELISA). RESULTS: Umbilical artery PI was significantly higher in women with IUGR than in those with pre-eclampsia, whereas uterine artery PI was not statistically significantly different. Maternal soluble fms-like tyrosine kinase-1 (sFlt-1) levels were higher in women with pre-eclampsia than in those with IUGR (P < 0.0001). Umbilical vein basic fibroblast growth factor (bFGF) levels were lower in women with pre-eclampsia than in those with IUGR (P < 0.05). Placental growth factor (PlGF) levels in the umbilical vein were below the detection limit in nearly all samples of IUGR fetuses and lower than in those with pre-eclampsia (P < 0.001). Maternal PlGF levels were inversely correlated with PI values of both vessels. In the umbilical vein sFlt-1 was positively and soluble kinase insert domain receptor (sKDR) negatively correlated with umbilical artery PI. No correlation could be found in the serum of the umbilical artery for all growth factors and for vascular endothelial growth factor (VEGF) in all compartments. CONCLUSIONS: The correlations between maternal and fetal angiogenic growth factor serum levels and Doppler ultrasound indices of uterine and umbilical arteries in pre-eclampsia and IUGR reflect the severity of the disorders especially for the fetus. A combination of both measurements may be useful in future screening for early prediction of pregnancy complications. Published by John Wiley & Sons, Ltd.     

子痫前期患者胎盘组织及外周血中胎盘生长因子及可溶性血管内皮生长因子受体1的表达及意义

目的 研究子痫前期患者胎盘生长因子(placental growth factor, PlGF)、可溶性血管内皮生长因子受体1(soluble vascular endothelial growth factor receptor 1, sFlt-1)的mRNA及蛋白质在胎盘组织中的表达及外周血中的浓度水平, 探讨其与子痫前期的关系。 方法 选取57例子痫前期患者（轻度子痫前期31例、重度子痫前期26例）和60例健康孕妇,用半定量RT-PCR及western blot分别检测胎盘组织中sFlt-1、PlGF的mRNA及蛋白质的表达水平, ELISA法检测血清中PlGF和sFlt-1水平。 结果 轻、重度子痫前期患者组胎盘组织PlGF mRNA、蛋白质表达水平以及血清PlGF浓度均显著低于健康孕妇组（t分别为14.22、21.80、12.10、15.17、7.14、10.18,P均<0.01）;而胎盘组织sFlt-1 mRNA、蛋白质表达水平以及血清sFlt-1浓度显著高于健康孕妇组（t分别为12.43、17.06、13.70、18.84、13.55、15.19,P均<0.01）;轻、重度子痫前期组外周血PlGF和sFlt-1之间呈显著负相关（r=-0.49, r=-0.53,P<0.05）。 结论 子痫前期胎盘组织中PlGF水平降低伴sFlt1水平升高与子痫前期的发病密切相关。     

Complement activation induces dysregulation of angiogenic factors and causes fetal rejection and growth restriction

Immune mechanisms have been implicated in placental dysfunction in patients with recurrent miscarriages and intrauterine growth restriction (IUGR), but the mediators are undefined. Here we show that complement activation, particularly C5a, is a required intermediary event in the pathogenesis of placental and fetal injury in an antibody-independent mouse model of spontaneous miscarriage and IUGR, and that complement activation causes dysregulation of the angiogenic factors required for normal placental development. Pregnancies complicated by miscarriage or growth restriction were characterized by inflammatory infiltrates in placentas, functional deficiency of free vascular endothelial growth factor (VEGF), elevated levels of soluble VEGF receptor 1 (sVEGFR-1, also known as sFlt-1; a potent anti-angiogenic molecule), and defective placental development. Inhibition of complement activation in vivo blocked the increase in sVEGFR-1 and rescued pregnancies. In vitro stimulation of monocytes with products of the complement cascade directly triggered release of sVEGFR-1, which sequesters VEGF. These studies provide the first evidence linking the complement system to angiogenic factor imbalance associated with placental dysfunction, and identify a new effector of immune-triggered pregnancy complications.     

Angiogenic profile and smoking in the Finnish Genetics of Pre-Eclampsia Consortium (FINNPEC) cohort

Objectives: The biological mechanism by which smoking reduces the risk of pre-eclampsia (PE) is unresolved. We studied serum levels of soluble fms-like tyrosine kinase 1 (sFlt-1), placental growth factor (PlGF) and their ratio, in addition to soluble endoglin (sEng) in early and late pregnancy to ascertain whether these factors are altered in women who smoke.

Subjects and methods: First trimester serum samples were available from 217 women who later developed PE and 238 women who did not develop PE. Second/third trimester serum samples were available from 174 PE and 54 non-PE women.

Results: PE women who smoked during pregnancy had elevated first trimester concentrations of serum PlGF [geometric mean (95% CI): 39.8 (32.6–48.5) pg/ml, p?=?.001] and reduced sEng concentration [5.0 (4.6–5.6) ng/ml, p?=?.047] compared to PE non-smokers [30.0 (28.1–32.1) pg/ml and 6.1 (5.9–6.4) ng/ml, respectively]. Non-smoking women in the PE group had the highest sFlt-1/PlGF ratio in early and late pregnancy.

Conclusions: The protective effect of smoking in reducing the risk of PE may be due to the early pregnancy change towards pro-angiogenic marker profile. Also, in late pregnancy, smoking exerted effect in sFlt-1/PlGF ratio in PE pregnancies, and may complicate its use as a prognostic and diagnostic marker.

• Key messages

• Smoking appears to have angiogenic effects in early pregnancy with reduced sEng concentrations and elevated PlGF concentrations in both normal and PE pregnancies.

• Throughout pregnancy, smoking exerted effect in PlGF concentration and sFlt-1/PlGF ratio in PE pregnancies, and thus may complicate its use as a prognostic and diagnostic marker.

     

Basic fibrobrast growth factor induces the secretion of vascular endothelial growth factor by human aortic smooth muscle cells but not by endothelial cells

BACKGROUND: Endothelial cell dysfunction and smooth muscle cell (SMC) proliferation are major events in atherogenesis. Both cells are a source of growth factors that mediate cellular proliferation and chemotaxis. Inappropriate production of, and/or response to, these growth factors (such as vascular endothelial growth factor, VEGF, and basic fibroblast growth factor (bFGF)) may contribute to atherogenesis and therefore to disease progression. METHODS: Production of VEGF and its soluble receptor (sFlt-1) by human SMCs and human umbilical endothelial cells (HUVECs) after stimulation with bFGF were examined by ELISA of cell culture media and by Western blotting. RESULTS: Smooth muscle cells produced significantly more VEGF than HUVECs (P<0.05) after 24 h of culture with bFGF levels > or =0.001 microg mL(-1). bFGF induced dose-dependent production of VEGF by SMCs, where maximum production was present in 1 microg mL(-1) of bFGF. Conversely, the SMCs produced less sFlt-1 than HUVECs (P<0.05). However, bFGF induced dose-dependent phosphorylation of Flt1 and another VEGF receptor, KDR, in HUVECs but not SMCs. There was no VEGF or sFLT-1 after 6 h of culture in any dose of bFGF in either type of cell. CONCLUSIONS: Differences in the production of VEGF and sFlt-1 by SMCs and HUVECs are consistent with the role of these cells in angiogenesis. Induction of VEGF production and expression by bFGF in these cells indicates that this growth factor may participate in angiogenesis indirectly by the induction of VEGF. The production of sFlt-1 by both cell types is in agreement with the notion that sFlt-1 may be involved in the regulation of VEGF activity. Additionally, the ability of bFGF to induce dose-dependent phosphorylation of KDR in HUVECs highlights the important role of bFGF in VEGF-mediated angiogenic processes.     

Pregnancy-induced hypertension and preeclampsia: levels of angiogenic factors in malaysian women

Preeclampsia (PE) is a major contributor to maternal and fetal mortality. The cause of preeclampsia remains unclear, but oxidative stress on the endothelium leading to endothelial dysfunction is said to be the root cause of the disease. The aim of this study was to measure and determine the plasma levels of key angiogenic factors in pregnancy as an indicator for the early onset of preeclampsia in pregnancy. Plasma levels of circulating a soluble fms like tyrosine kinase-1 (sFlt-1), an anti-angiogenic factor, vascular endothelial growth factor (VEGF) and placental growth factor (PIGF), both pro-angiogenic factors were analyzed in normal pregnant Malaysian women (control group, n = 34), women with pregnant induced hypertension (PIH, n = 34) and women with preeclampsia (PE, n = 34) all at three gestational ages, 24–28 weeks (early pregnancy: EP), 32–36 weeks (late pregnancy: LP) and 6 weeks after delivery (postpartum: PN). The plasma levels of angiogenic factors were determined by ELISA. sFlt-1 levels were elevated in PIH and PE patients as compared to controls. PIGF and VEGF were significantly decreased in PIH and PE as compared to the controls. These results suggest that elevated concentration of sFlt-1 and suppressed levels of PIGF and VEGF may contribute to the development of hypertension in pregnancy which precedes preeclampsia.     

Abnormal maternal cardiac function and morphology in pregnancies complicated by intrauterine fetal growth restriction.

OBJECTIVE: To explore maternal cardiac function through an echocardiographic evaluation, in a group of nulliparous patients with intrauterine fetal growth restriction during the third trimester of pregnancy. METHODS: Twenty-one consecutive nulliparous pregnant women who had fetuses with intrauterine growth restriction (IUGR) and abnormal umbilical artery Doppler pulsatility index (PI) underwent maternal echocardiographic examination during the third trimester of gestation. The data were then compared with those obtained from 21 normal nulliparous women who had fetuses with an estimated fetal weight > 10th percentile and a normal umbilical artery Doppler PI who were considered as the control group. RESULTS: Heart rate was slightly lower in the IUGR group, whereas blood pressure and total vascular resistance were higher compared with the control subjects. End-diastolic volume, stroke volume and cardiac output were lower in the IUGR patients compared with normal patients. The IUGR group had smaller left atrial maximal dimensions and greater left atrial minimal areas compared with the control subjects. Left atrial function was depressed in the IUGR group. A smaller left ventricular mass was present in the IUGR patients compared with the control subjects. Isovolumetric relaxation time (IVRT) was prolonged in the IUGR patients compared with the controls. CONCLUSIONS: The absence of a 'correct' maternal cardiovascular compensatory response to abnormal trophoblastic invasion, might be one of the factors that slowly determine the conditions of reduced placental perfusion and eventually of the development of fetal growth restriction.     

Serum heat shock protein 70 levels in relation to circulating cytokines, chemokines, adhesion molecules and angiogenic factors in women with preeclampsia

BackgroundWe have previously reported that serum levels of 70 kDa heat shock protein (Hsp70, HSPA1A) are increased and reflect systemic inflammation, oxidative stress and hepatocellular injury in preeclampsia. The purpose of this study was to determine whether increased serum Hsp70 concentrations in women with preeclampsia are related to circulating levels of cytokines, chemokines, adhesion molecules and angiogenic factors, the key players in the pathogenesis of the disease.MethodsSixty preeclamptic patients and 60 normotensive, healthy pregnant women were involved in this case-control study. Levels of Hsp70 (HSPA1A) and transforming growth factor (TGF)-beta1 in maternal sera were assessed by ELISA. Serum levels of interleukin (IL)-1beta, IL-1 receptor antagonist, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p40, IL-12p70, IL-18, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interferon-gamma-inducible protein (IP)-10, monocyte chemotactic protein (MCP)-1, intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 were determined by multiplex suspension array. Serum total soluble fms-like tyrosine kinase-1 (sFlt-1) and biologically active placental growth factor (PlGF) levels were measured by electrochemiluminescence immunoassay. For statistical analyses, the Mann–Whitney U-test, the Fisher exact and Pearson chi-square tests, the Spearman rank order correlation, multiple linear regression and logistic regression were applied.ResultsSerum levels of Hsp70 were significantly higher in preeclamptic patients than in healthy pregnant women. Additionally, most of the measured inflammatory variables differed significantly between the two study groups except for serum IL-1beta and TGF-beta1 levels and IL-18/IL-12p70 and IL-12p70/IL-12p40 ratios, indicating a bias toward a pro-inflammatory status in preeclampsia. Preeclamptic patients had significantly higher sFlt-1 levels and sFlt-1/PlGF ratio and significantly lower PlGF concentrations as compared to healthy pregnant women. In the preeclamptic group, serum Hsp70 concentrations showed significant correlations with serum levels of IL-12p40 (R = 0.59, p < 0.001), MCP-1 (R = 0.43, p < 0.001), ICAM-1 (R = 0.39, p = 0.0020) and VCAM-1 (R = 0.46, p < 0.001). Furthermore, elevated serum Hsp70 level and sFlt-1/PlGF ratio had a synergistic (joint) effect in the risk of preeclampsia, as shown by the substantially higher odds ratios of their combination than of either alone.ConclusionsIncreased serum Hsp70 concentrations in women with preeclampsia were associated with pro-inflammatory changes in circulating cytokine profile, suggesting that circulating Hsp70 might contribute to the development of the excessive systemic inflammatory response characteristic of the maternal syndrome of the disease.     

High maternal blood S100B concentrations in pregnancies complicated by intrauterine growth restriction and intraventricular hemorrhage

BACKGROUND: Intrauterine growth restriction (IUGR) is associated with perinatal mortality and with neurologic damage from intraventricular hemorrhage (IVH). We investigated whether S100B, a neural protein found in high concentrations after cell injury in the nervous system, is increased in serum of women whose pregnancies are complicated by IUGR and whose newborns develop IVH. We also explored the prognostic accuracy of maternal serum S100B for IVH in the newborn. METHODS: We conducted a case-control study of 106 pregnancies complicated by IUGR, including a subgroup (n = 26) who developed IVH after birth, and 212 unaffected pregnancies matched for gestational age. Ultrasound examination, Doppler velocimetry patterns (in the utero-placental vessels and middle cerebral artery), and maternal blood collection were performed before birth; cerebral ultrasound and neurologic examinations were performed after birth. RESULTS: S100B was higher (P <0.001) in IUGR pregnancies complicated by IVH than in those that were not and in controls. At a cutoff of 0.72 microg/L, sensitivity was 100% [95% confidence interval (95% CI), 87%-100%] and specificity was 99.3% (97.5%-99.9%) for prediction of IVH (area under the ROC curve, 0.999). The prevalence of IVH was 8.2% in the whole study population, 93% (95% CI, 83.6%-100%) in those with maternal S100B >0.72 microg/L, and 0% (0%-2.5%) in those with maternal S100B <0.72 microg/L. CONCLUSION: For prediction of IVH, measurements of maternal S100B may be useful at times before clinical, laboratory, and ultrasound patterns can identify risk of IVH.     

The Role of Angiogenic and Antiangiogenic Factors in the Second Trimester in the Prediction of Preeclampsia in Pregnant Women With Type 1 Diabetes

OBJECTIVE

To assess the association between circulating angiogenic and antiangiogenic factors in the second trimester and risk of preeclampsia in women with type 1 diabetes.

RESEARCH DESIGN AND METHODS

Maternal plasma concentrations of placental growth factor (PlGF), soluble fms-like tyrosine kinase 1 (sFlt-1), and soluble endoglin (sEng) were available at 26 weeks of gestation in 540 women with type 1 diabetes enrolled in the Diabetes and Preeclampsia Intervention Trial.

RESULTS

Preeclampsia developed in 17% of pregnancies (n = 94). At 26 weeks of gestation, women in whom preeclampsia developed later had significantly lower PlGF (median [interquartile range]: 231 pg/mL [120–423] vs. 365 pg/mL [237–582]; P < 0.001), higher sFlt-1 (1,522 pg/mL [1,108–3,393] vs. 1,193 pg/mL [844–1,630] P < 0.001), and higher sEng (6.2 ng/mL [4.9–7.9] vs. 5.1 ng/mL[(4.3–6.2]; P < 0.001) compared with women who did not have preeclampsia. In addition, the ratio of PlGF to sEng was significantly lower (40 [17–71] vs. 71 [44–114]; P < 0.001) and the ratio of sFlt-1 to PlGF was significantly higher (6.3 [3.4–15.7] vs. 3.1 [1.8–5.8]; P < 0.001) in women who later developed preeclampsia. The addition of the ratio of PlGF to sEng or the ratio of sFlt-1 to PlGF to a logistic model containing established risk factors (area under the curve [AUC], 0.813) significantly improved the predictive value (AUC, 0.850 and 0.846, respectively; P < 0.01) and significantly improved reclassification according to the integrated discrimination improvement index (IDI) (IDI scores 0.086 and 0.065, respectively; P < 0.001).

CONCLUSIONS

These data suggest that angiogenic and antiangiogenic factors measured during the second trimester are predictive of preeclampsia in women with type 1 diabetes. The addition of the ratio of PlGF to sEng or the ratio of sFlt-1 to PlGF to established clinical risk factors significantly improves the prediction of preeclampsia in women with type 1 diabetes.Preeclampsia is characterized by the development of hypertension and new-onset proteinuria during the second half of pregnancy (1,2), leading to increased maternal morbidity and mortality (3). Women with type 1 diabetes are at increased risk for development of preeclampsia during pregnancy, with rates being two-times to four-times higher than that of the background maternity population (4,5). Small advances have come from preventive measures, such as low-dose aspirin in women at high risk (6); however, delivery remains the only effective intervention, and preeclampsia is responsible for up to 15% of preterm births and a consequent increase in infant mortality and morbidity (7).Although the etiology of preeclampsia remains unclear, abnormal placental vascular remodeling and placental ischemia, together with maternal endothelial dysfunction, hemodynamic changes, and renal pathology, contribute to its pathogenesis (8). In addition, over the past decade accumulating evidence has suggested that an imbalance between angiogenic factors, such as placental growth factor (PlGF), and antiangiogenic factors, such as soluble fms-like tyrosine kinase 1 (sFlt-1) and soluble endoglin (sEng), plays a key role in the pathogenesis of preeclampsia (8,9). In women at low risk (10–13) and women at high risk (14,15), concentrations of angiogenic and antiangiogenic factors are significantly different between women who later develop preeclampsia (lower PlGF, higher sFlt-1, and higher sEng levels) compared with women who do not.Few studies have specifically focused on circulating angiogenic factors and risk of preeclampsia in women with diabetes, and the results have been conflicting. In a small study, higher sFlt-1 and lower PlGF were reported at the time of delivery in women with diabetes who developed preeclampsia (16). In a longitudinal prospective cohort of pregnant women with diabetes, Yu et al. (17) reported increased sFlt-1 and reduced PlGF in the early third trimester as potential predictors of preeclampsia in women with type 1 diabetes, but they did not show any difference in sEng levels in women with preeclampsia compared with women without preeclampsia. By contrast, Powers et al. (18) reported only increased sEng in the second trimester in women with pregestational diabetes who developed preeclampsia.The aim of this study, which was significantly larger than the previous studies highlighted, was to assess the association between circulating angiogenic (PlGF) and antiangiogenic (sFlt-1 and sEng) factors and the risk of preeclampsia in women with type 1 diabetes. A further aim was to evaluate the added predictive ability and clinical usefulness of angiogenic factors and established risk factors for preeclampsia risk prediction in women with type 1 diabetes.     

彩色多谱勒对IUGR脐动脉血流的检测

目的：应用彩色多谱勒检测正常妊娠和胎儿宫内发育迟缓（以简称ＩＵＧＲ）的脐动脉血流指标ＰＩ、ＲＩ、Ｓ／Ｄ值。材料和方法：应用彩色多谱勒检测了２００例正常妊娠和１１１例ＩＵＧＲ的脐动脉血流,孕周从２６－４１周。结果：正常妊娠组及ＩＵＧＲ组的脐动脉血流指标均随妊娠周数的增高而降低,但ＩＵＧＲ组的脐动脉血流ＰＩ、ＲＩ、Ｓ／Ｄ值明显高于正常妊娠组（Ｐ〈０．０１）。孕３０周后,脐动脉血流Ｓ／Ｄ值〉４,揭示胎儿预后不良。结论：脐动脉血流检测不仅是胎儿监护的一种方法,而且可作为诊断ＩＵＧＲ的一个指标及对估计胎儿预后有一定的临床价值。     

Maternal cardiac function in fetal growth-restricted and non-growth-restricted small-for-gestational age pregnancies.

OBJECTIVE: To compare maternal cardiac function in women with intrauterine growth restriction (IUGR) to those with small-for-gestational age (SGA) pregnancies (non-IUGR). METHODS: This was a cross-sectional study involving maternal echocardiography and uterine, umbilical and fetal middle cerebral artery Doppler assessment in 52 normotensive women at 20-36 weeks' gestation with SGA fetuses (26 IUGR and 26 non-IUGR). RESULTS: In the IUGR (compared to the non-IUGR) group, maternal cardiac output (CO) was lower (4.7 vs. 6.1 L/min, P<0.001) and total vascular resistance (TVR) was higher (1444 vs. 1088 dynes/s/cm5, P<0.001). The lower CO was due to a lower preload, demonstrated by a reduced stroke volume (59.9 vs. 73.6 mL, P<0.01) and smaller left atrial diameter (LAD) (31.5 vs. 34.1 mm, P=0.01). Mean arterial pressure and diastolic function were similar between the groups. Logistic regression and receiver-operating characteristics curve analysis for detection of IUGR demonstrated that a model using TVR, LAD, fetal middle cerebral artery pulsatility index and gestational age, had a sensitivity of 96.2% and a specificity of 84.6%. CONCLUSIONS: Maternal echocardiography can provide a very sensitive tool for identifying IUGR pregnancies.     

彩色多普勒超声对宫内发育迟缓胎儿动静脉的血流动力学研究

目的应用彩色多普勒超声检测宫内发育迟缓(IUGR)胎儿动静脉的血流动力学参数。方法测量胎儿静脉导管和脐静脉的血流量,计算静脉导管(DV)分流率;测量胎儿脐动脉、大脑中动脉(MCA)、肾动脉的搏动指数(PI)、阻力指数(RI)及脐动脉收缩期最大流速与舒张末期流速的比值(S/D),结果进行对比分析。结果与对照组比较,IUGR组胎儿脐动脉的PI、RI、S/D及肾动脉的RI显著增高,MCA的PI、RI及S/D显著减低,DV分流率显著增加,差异均有统计学意义(P<0.05)。结论彩色多普勒超声通过综合监测胎儿血流动力学变化,可以全面、客观地评价胎儿的供血情况,预测胎儿宫内缺氧和IUCR情况。     

Absolute and relative quantification of placenta-specific micrornas in maternal circulation with placental insufficiency-related complications

Placental insufficiency-related complications are one of the leading causes of maternal and perinatal morbidity and mortality. This study investigated the quantification of placenta-specific microRNAs (miRNAs) in the maternal circulation during gestation in a cohort of women with normally progressing pregnancies, the differentiation between placental insufficiency-related complications and normally progressing pregnancies, and the differentiation between placental insufficiency and normally progressing pregnancies during the early stages of gestation. Both absolute and relative quantification of placenta-specific miRNAs (ie, miR-516-5p, miR-517*, miR-518b, miR-520a*, miR-520h, miR-525, and miR-526a) was determined in 50 women with normally progressing pregnancies, 32 with complicated pregnancies [21 with preeclampsia with or without intrauterine growth retardation (IUGR) and 11 with IUGR], and 7 women with pregnancies at various gestational stages who later developed preeclampsia and/or IUGR using real-time PCR and a comparative C(T) method relative to normalization factor (ie, geometric mean of ubiquitous miR-16 and let-7d). Both quantification approaches revealed significant increases in extracellular placenta-specific miRNA levels over time in women with normally progressing pregnancies; however, they were not able to differentiate between normally progressing and complicated pregnancies at the time of preeclampsia and/or IUGR onset. Nevertheless, significant elevation of extracellular miRNA levels was observed during early gestation (ie, within the 12th to 16th weeks) in pregnancies with later onset of preeclampsia and/or IUGR. Early gestation extracellular miRNA screening can differentiate between women with normally progressing pregnancies and those who may later develop placental insufficiency-related complications.     

Correlation of Doppler and placental immunohistochemical features in normal and intrauterine growth-retarded fetuses.

The aim of this study was to correlate and compare Doppler and anatomical placental findings obtained from 48 normal and 35 intrauterine growth-retarded (IUGR) fetuses. The IUGR group consisted of 19 fetuses from pregnancies complicated by pre-eclampsia and 16 from healthy mothers. Color Doppler evaluation of umbilical, spiral and uterine arteries was performed. Placental specimens from both normal and growth-retarded fetuses were obtained at the time of delivery. Placental specimens were evaluated using histochemical and immunohistochemical techniques. A progressive decrease in the pulsatility index was observed in umbilical, spiral and uterine arteries throughout pregnancy in the normal-growth fetuses. High umbilical artery pulsatility index values were obtained in 29 out of the 35 growth-retarded fetuses, six of them showing absent or reversed end-diastolic umbilical artery flow pattern. A total of 13 IUGR fetuses showed high resistance uterine artery flow velocity waveforms. Increased pulsatility index values were obtained from the spiral arteries of 16 growth-retarded fetuses. Abnormal histological and histochemical placental patterns were observed in all the growth-retarded fetuses with umbilical artery Doppler abnormalities. The presence of a peculiar dendritic cell subpopulation, strongly resembling the Langerhans cells, expressing the HLA-DR+/CD1+ phenotype, was detected in all growth-retarded fetuses, whether there was maternal pathology or not. Our data show uterine and spiral artery data as being ineffective in the monitoring of IUGR fetuses. The placental extracellular matrix seems to play an important role in the regulation of the umbilical circulation. The presence of CD1+ cells as a sign of a possible immunological mechanism in the pathogenesis of the intrauterine growth retardation is discussed.     

Angiogenic growth factors in the diagnosis and prediction of pre-eclampsia

The pathogenesis of pre-eclampsia is still not completely known; however, in the recent decade, there have been tremendous research efforts leading to impressive results highlighting the role of a disturbed angiogenic balance as one of the key features of the disease. Numerous studies have shown the key role of the placenta in the pathogenesis of pre-eclampsia. A shift in the sFlt-1 (soluble Fms-like tyrosine kinase-1)/PlGF (placental growth factor) ratio is associated with the disease. Although pre-eclampsia seems to be a clearly defined disease, clinical presentation, and particularly the dynamics of the clinical course, can vary enormously. The only available tools to diagnose pre-eclampsia are blood pressure measurement and urine protein sampling. However, these tools have a low sensitivity and specificity regarding the prediction of the course of the disease or maternal and perinatal outcomes. The only cure for the disease is delivery, although a timely diagnosis helps in decreasing maternal and fetal morbidity and mortality. The sFlt1/PlGF ratio is able to give additional valuable information on the status and progression of the disease and is apt to be implemented in the diagnostic algorithm of pre-eclampsia. In the present review, we aim to provide an overview of the vast literature on angiogenesis and anti-angiogenesis factors in pre-eclampsia that have been published over the last decade. We introduce work from basic research groups who have focused on the pathophysiological basis of the disease. Furthermore, we review studies with a clinical focus in which the sFlt-1/PlGF ratio has been analysed along with other candidates for routine clinical assessment of pre-eclampsia.     

Altered angiogenesis in preeclampsia: evaluation of a new test system for measuring placental growth factor.

BACKGROUND: Decreased concentrations of the circulating angiogenic factors, free placental growth factor (PLGF) and free vascular endothelial growth factor (VEGF), and increased concentrations of the anti-angiogenic factor, soluble fms-like tyrosine kinase 1 (sFLT-1) have been observed during clinical preeclampsia. We established a new PLGF-ELISA kit for the measurement of PLGF in sera. In the present study, we demonstrated the assay characteristics by measurement of PLGF expression in normal and preeclamptic pregnancies as compared to an established research kit. METHODS: Blood samples were taken from 64 women with singleton uncomplicated pregnancies for longitudinal measurement of PLGF in the course of pregnancy. In 30 preeclamptic patients, serum levels of PLGF and sFLT-1 were measured by Human PLGF-ELISA and Human sVEGF R1 ELISA according to the described test principles. The assay characteristics of the new PLGF-ELISA were determined and the results were compared to those performed with an available research kit. RESULTS: The PLGF concentration in normal pregnancies showed a steady increase starting at the beginning of the second trimester with a peak at 28-32 weeks and a consistent decline thereafter. The preeclamptic pregnancies had significant lower serum concentrations of PLGF and significant higher serum concentrations of sFLT-1 as compared to the non-preeclamptic pregnancies. All the measured assay characteristics fulfilled the required specifications. Comparison of the values of the new PLGF-ELISA and the established research kit resulted in a correlation coefficient of 0.921. CONCLUSIONS: Our results support the hypothesis that an imbalance between factors promoting angiogenesis, such as PLGF, and factors antagonizing angiogenesis, such as sFLT-1, has a fundamental role in the pathogenesis of preeclampsia. The new established ELISA test can be considered reliable and it offers many advantages. As it is authorized for routine diagnostic testing, it may offer new possibilities in the prediction of preeclampsia in clinical routine.     

甲状腺激素及瘦素水平变化对胎儿生长发育的影响

目的　探讨甲状腺激素和瘦素水平对胎儿生长发育的影响。方法　采用放射免疫法测定 2 4例 IUGR孕妇 (受试组 )、4 0例正常孕妇 (对照组 )血清及其新生儿脐血中的甲状腺激素 [游离 T3(FT3)、游离 T4 (FT4 )、促甲状腺素(TSH) ]和瘦素水平 ,分析甲状腺激素、瘦素水平与胎儿生长发育之间的关系。结果　 IUGR孕妇血清及脐血中 TSH水平均高于对照组 ,而 FT4 及瘦素水平均低于对照组 (P<0 .0 5 ) ;脐血 FT3、FT4 及瘦素水平均与胎儿出生体重呈正相关(P<0 .0 5、P<0 .0 1)。孕妇血清瘦素水平与脐血瘦素水平无相关。瘦素水平与 TSH呈负相关。结论　胎儿生长发育与脐血 TSH、FT3、FT4 及瘦素水平均有密切关系 ,对评估胎儿体重和生长发育均具有重要的临床意义。     

促血管生成素-2与胎儿生长受限发病的关系

目的:探讨促血管生成素-2(Ang-2)在母血、脐血血清中的水平和在胎盘组织上的表达与胎儿生长受限(FGR)发病的关系。方法:选取剖宫产分娩足月FGR患儿的孕妇30例作为实验组,同期剖宫产分娩正常足月婴儿的孕妇30例作为对照组。采用酶联免疫法(ELISA)测定母血和脐血血清中Ang-2的水平;采用免疫组织化学方法(SP法)检测孕妇胎盘组织中Ang-2的表达。结果:(1)实验组和对照组母血血清中Ang-2的水平分别为(8.86±0.43)ng/mL、(19.31±0.25)ng/mL。实验组和对照组脐血血清中Ang-2的水平分别为(10.92±0.41)ng/mL、(22.28±0.23)ng/mL;实验组和对照组胎盘组织中Ang-2的表达分别为118.17±1.59、151.64±1.64。实验组母血和脐血血清中Ang-2的水平和胎盘组织中Ang-2的表达均显著低于对照组(P<0.01)。(2)实验组胎盘组织中Ang-2的表达与母血血清Ang-2水平呈正相关(r=0.93,P<0.01);实验组胎盘组织中Ang-2的表达与脐血血清Ang-2水平呈正相关(r=0.93,P<0.01);母血血清中Ang-2水平与脐血血清中Ang-2水平呈正相关(r=0.99,P<0.01)。结论:母血和脐血血清中Ang-2水平均降低和胎盘组织中Ang-2表达降低可能参与了FGR的发病过程。     

Multivariable analysis of tests for the diagnosis of intrauterine growth restriction.

OBJECTIVES: To describe how data from antenatal fetal ultrasound biometry, amniotic fluid index and umbilical artery Doppler can be appropriately combined using multivariable models and to investigate how the addition of these ultrasound parameters influences the ability to predict intrauterine growth restriction (IUGR). METHODS: This was a prospective cohort study involving 274 low-risk pregnancies undergoing serial ultrasound examination at predetermined intervals. Standard deviation (Z) scores of the last values for fetal abdominal area (FAA), growth velocity of the FAA, amniotic fluid index (AFI) and umbilical artery Doppler pulsatility index prior to delivery were calculated for 260 fetuses. Customized estimated fetal weight (cEFW) centiles were also calculated using the last EFW before delivery after adjustment for fetal gender, gestational age, birth order and maternal weight, height and ethnic origin. Following delivery the neonatal ponderal index was calculated and centile position obtained. A neonatal ponderal index <25(th) centile served as the main outcome measure for diagnosis of IUGR. Logistic regression analysis was used to delineate the predictive value of the three fetal growth tests FAA, FAA growth velocity and cEFW and the additional values of AFI and pulsatility index of the umbilical artery. RESULTS: The areas under the receiver-operating characteristics (ROC) curves (95% confidence interval) for FAA, FAA growth velocity and cEFW alone were 0.819 (0.748-0.891), 0.784 (0.699-0.869) and 0.74 (0.643-0.837), respectively, in the prediction of a neonatal ponderal index <25(th) centile. The addition of both the AFI and pulsatility index to FAA, FAA growth velocity and cEFW generated small increases in the areas, to 0.831 (0.758-0.904), 0.817 (0.735-0.899) and 0.766 (0.672-0.859), respectively. These improvements in diagnostic prediction were not statistically significant. CONCLUSIONS: The addition of AFI and umbilical artery pulsatility index to the fetal biometry parameters did not significantly increase the ROC areas in the study population. The approach applied in this study is useful in the context of hypothesis generation. Further studies using larger data sets and other predictors should be carried out using the analytical techniques outlined in this paper to determine the contribution of various antenatal tests in the prediction of IUGR.