妊娠晚期特发性羊水量异常羊水的生化改变探讨

目的:探讨妊娠晚期特发性羊水量异常者的羊水生化改变.方法:对73例妊娠晚期特发性羊水异常患者(59例特发性羊水过少、14例特发性羊水过多)进行羊水10项生化指标检测和分析,与同期随机选取的50例无妊娠合并症和并发症的单胎足月孕妇羊水进行比较.结果:特发性羊水过少组羊水中钠浓度、钙浓度、葡萄糖及渗透压较正常组降低,肌酐浓度升高,其差异有统计学意义(P<0.05);特发性羊水过多组各项羊水生化参数与羊水量正常组差异无统计学意义(P>0.05).结论:妊娠晚期特发性羊水过少者羊水中的钠浓度、钙浓度、葡萄糖及渗透压有改变.     

Antiviral activity of the human amniotic fluid and placenta

Antiviral activity (3-24 IU/ml) was detected in the amniotic fluid of 21 of 25 pregnant women examined (84%) by conventional interferon assay using human amniotic cells (WISH) and vesicular stomatitis virus. The pregnancies were from 16 to 42 weeks gestation without obvious viral infection or other complications. The antiviral substance in the amniotic fluid seems to be interferon alpha, because it was not inactivated by heat (56 degrees C, 30 min) or acid (pH 2.0, 24 hrs.) and it was inactivated by anti-human interferon alpha neutralizing serum. Neither the incidence nor the titer of the activity in amniotic fluid has any relationship with gestational age, age of the pregnant women or parity. Antiviral activity (4-32 IU/10 mg protein/ml) was also detected in the placenta homogenate of 23 of 25 pregnant women examined (92%). The pregnancies were from 7 to 41 weeks of gestation with or without complication. The titer of antiviral activity was relatively high in the placenta of babies with central nervous system anomalies and it was low in the placenta of intrauterine growth retardation pregnancies, compared with normal pregnancies. The activity in the placenta was not inactivated by heat (56 degrees C, 30 min), acid (pH 2.0, 24 hrs.), anti-human interferon alpha neutralizing anti-serum or anti-human interferon beta neutralizing anti-serum.     

改良荧光原位杂交技术在产前诊断中的应用

目的:评价改良荧光原位杂交(fluorescent in situ hybridization,FISH)技术在产前诊断中的应用。方法:用改良FISH技术检测119例孕16~24周孕妇的羊水间期细胞及10例孕25~32周胎儿脐血间期细胞,5例孕9~12周绒毛间期细胞,每例均行常规染色体核型分析。结果:应用改良FISH法,所有样本均在6h内获得检测结果,除2例羊水培养失败外,其余样本均在3周内获得细胞遗传学诊断。两种方法均检出特氏综合征、18-三体综合征、21-三体综合征各1例,另5例常规染色体核型分析异常,因超出检测范围,FISH法未能检出,所有样本的两种方法检测结果均一致。结论:经改良后的FISH技术缩短了诊断时间,缓解了孕妇及家属的焦虑心情,且可用于多种不同样本的检测,因其高效、省时、取材多样等优点在产前诊断具有重要的临床价值。     

Comparative study of proteinase inhibitors in human amniotic fluid and maternal serum

Serum and amniotic fluid samples from five pregnant women were analyzed for different proteinase inhibitors by ion-exchange chromatography on DEAE-cellulose. Alpha-2 macroglobulin was found to be absent in amniotic fluid. Even though alpha-1 antichymotrypsin was identified in amniotic fluid, its concentration gradient between serum and amniotic fluid (20--60) was much higher than that of alpha-1 antitrypsin (8--20). Two forms of alpha-1 antitrypsin were identified in both the fluids. In pregnancy, the ratio of the two forms was altered in favour of the less anionic form compared to normal sera. The contribution of the heat stable proteinase inhibitor for total antitryptic activity in amniotic fluid was more than in serum. The observation that urine samples from immature infants had higher heat stable inhibitory activity than urine samples from normal infants suggests that the heat stable inhibitor in amniotic fluid may arise from fetal kidney.     

早孕妇女血清及蜕膜组织液对自然杀伤细胞活性及淋巴细胞增殖的影响

目的　探讨早孕妇女血清及蜕膜组织液对自然杀伤 (NK)细胞及淋巴细胞功能的影响。方法　采用同位素掺入法 ,检测 3 2例正常早孕妇女血清 (早孕血清组 )及蜕膜组织液 (蜕膜组织液组 )对正常妇女外周血中NK细胞活性 (% )及淋巴细胞增殖功能的影响 ,并采用 2 5例正常未孕妇女的血清作对照 (对照组 )。结果　早孕血清组、蜕膜组织液组及对照组的NK细胞活性分别为 (3 5 .8±3 .0 ) %、(19.1± 2 .9) %及 (46.1± 3 .2 ) % ,淋巴细胞增殖功能 [每分钟同位素放射次数 (CPM) ]分别为(3 689± 4 5 6)CPM、(2 0 5 9± 2 88)CPM及 (4883± 64 3 )CPM ,3者间差异均有极显著性 (P <0 .0 1) ,且蜕膜组织液组均显著低于早孕血清组 (P <0 .0 0 5 )。结论　正常早孕妇女血清及蜕膜组织液具有免疫抑制作用 ,而蜕膜组织液可能是调节早期妊娠子宫局部免疫功能的重要因素之一。     

The arylsulphatase A (EC 3.1.6.1) activity in serum, urine and amniotic fluid from pregnant women with EPH-gestosis

In serum, urine and amniotic fluid obtained from the 52. women divided into three groups the arylsulphatase A (EC 3.1.6.1) activity was measured by the modified Lee-Vaupel and Conzelmann method. It was noticed in serum from the pregnant women with EPH-gestosis the statistically significant (p < 0.05) increase in the enzyme activity comparing to the results from non-pregnant women and pregnant with normal course of pregnancy. It was no statistically significant differences in the urine from pregnant with EPH-gestosis and from the healthy pregnant, but there was the increase (p < 0.01) in the enzyme activity in amniotic fluid from pregnant women with EPH-gestosis comparing to the physiological course of pregnancy. According to our data, the arylsulphatase A activity assay could be recommended as a diagnostic marker in the EPH-gestosis.     

Amniotic fluid and maternal serum leptin levels in pregnant women who subsequently develop preeclampsia

OBJECTIVES: To study the correlation between amniotic fluid leptin levels and maternal serum leptin levels during the early second trimester, and to determine whether the ratios of amniotic fluid leptin levels to maternal serum leptin levels are elevated in pregnant women who subsequently develop preeclampsia. STUDY DESIGN: Samples from 120 pregnant women were included in this prospective study, of which 20 were from pregnant women who subsequently developed preeclampsia and 100 were from normal pregnant women. Both the amniotic fluid and the maternal serum leptin levels were ascertained by radioimmunoassay (RIA). RESULTS: A strong correlation between amniotic fluid leptin levels and maternal serum leptin levels was observed in both preeclamptic and normal pregnant women. In addition, the ratios of amniotic fluid leptin levels to maternal serum leptin levels were positively correlated to amniotic fluid leptin levels, but negatively correlated to maternal serum leptin levels. Furthermore, the ratios of amniotic fluid leptin levels to maternal serum leptin levels in preeclamptic women were significantly higher than those in normal pregnant women. CONCLUSIONS: Amniotic fluid leptin levels correlated with maternal serum leptin levels during the early second trimester. The ratios of amniotic fluid leptin levels to maternal serum leptin levels were elevated in preeclamptic women. However, the maternal serum leptin levels themselves showed no such elevation. Therefore, this elevated ratio may be a marker at the early stage of pregnancy in preeclamptic women.     

Immunomodulating properties of sera in normal pregnant women]

The purpose of this study was to analyze the immunosuppressive properties of sera of normal pregnant women. Effects of pregnancy sera on the proliferation of mitogen (PHA, Con A, PWM)-induced lymphocytes were examined in healthy primiparas. The sera of normal pregnant women inhibited the proliferation of PHA- and Con A- induced both autologous and allogenic lymphocytes (obtained from nonpregnant and pre-eclamptic women), while remaining neutral towards the proliferation of PWM- induced lymphocytes in comparison with fetal calf serum. The basic immunomodulatory activity of the sera in pregnant women was contained in the heat-stable fraction.     

Pregnancy-associated major basic protein in amniotic fluid.

The pregnancy-associated major basic protein, a protein elevated in the sera of all pregnant women, is virtually identical to the eosinophil granule major basic protein. To determine whether pregnancy-associated major basic protein is present in amniotic fluid, we examined samples from both early and late gestation by a double antibody radioimmunoassay. A total of 112 amniotic fluids were tested and all but three contained levels of pregnancy-associated major basic protein greater than 400 ng/ml. Amniotic fluid pregnancy-associated major basic protein antigenic activity was immunochemically identical to that of the eosinophil granule major basic protein and also had identical physicochemical properties such as heat stability and the need for reduction and alkylation. Although the majority of amniotic fluid samples (90 of 112) were obtained from healthy women with normal gestations, the remaining 21 amniotic fluid samples were from women with Rh sensitization and from one gestation complicated by intrauterine growth retardation.     

Epidermal growth factor in urine of pregnant women and in amniotic fluid throughout pregnancy

To investigate the role of epidermal growth factor (EGF) in feto-placental development, we measured the urinary and amniotic fluid EGF levels throughout pregnancy. Thirty urinary samples of non-pregnant women, 85 of normal pregnant women, 21 of women with toxemic pregnancy, 17 of postpartum women and 30 of newborns, and 55 amniotic fluid samples of pregnant women with a variety of conditions necessitating amniotomy and amniocentesis at 25-39 weeks of gestation were collected. EGF concentrations were measured by double-antibody radioimmunoassay. Urinary EGF levels of pregnant women reached their peak (24.6 +/- 6.7 ng/mg creatinine) at 19-22 gestational weeks; after that, they slightly decreased. Although there is no significant difference between the urinary EGF levels of non-pregnant women (19.0 +/- 5.1) and those of pregnant women (18.1 +/- 3.2), the EGF levels of toxemic women (12.2 +/- 1.5) were lower than those of normal pregnant women. The levels in puerperium women were similar to those found during pregnancy. However, the neonates had higher urinary EGF concentrations than those in pregnant women. On the other hand, EGF levels in amniotic fluid were higher according to gestational weeks and the levels of intrauterine growth retardation (IUGR) cases lower compared with normal pregnancy. Furthermore, EGF concentrations in amniotic fluid have a significant correlation with the creatinine levels in amniotic fluid. These data suggest that EGF plays an important role in fetoplacental development and it is possible that the measurement of amniotic fluid EGF might become available for the clinical assessment of fetal maturation.     

Diagnostic significance of index alpha-amylase/glucose in amniotic fluid in prediction of fetal maturity.]

An increase in alpha-amylase activity with parallel decrease in glucose concentration in amniotic fluid is observed during pregnancy. This interdependence is a theoretical basis for using an alpha-amylase/glucose index in fetal maturity evaluation. The aim of the study was to investigate usefulness of the alpha-amylase/glucose index in amniotic fluid in prenatal fetal maturity diagnosis. The study was carried out on 180 pregnant women, chosen by random selection, hospitalized in Polish Mother's Health Centre Hospital in the period from 15.06.1994 to 31.12.1995. 223 samples of amniotic fluid were tested for glucose concentration and alpha-amylase activity. It was found that the alpha-amylase/glucose < 6.0 index indicates a possibility of RDS occurring in neonates born before 72 hours of performed determination. The alpha-amylase/glucose > or = 6.0 index has high diagnostic value (95.8%) in prenatal prediction of fetal lung maturity.     

Amniotic fluid glucose values in normal and abnormal pregnancies

Glucose values were determined in 102 urine samples of newborn infants and in 2295 amniotic fluid (AF) samples of women between the 14th and 42nd week of pregnancy. One thousand, six hundred fifty-five of the AF samples derived from normal pregnancies, 50 from pregnancies with fetal malformations, 115 from cases of hydramnios, 246 from pregnant women with an abnormal oral glucose tolerance test, and 230 from insulin-dependent diabetics. Mean AF glucose concentration rises slightly between the 14th and 17th week of pregnancy, decreasing from 46 to about 16 mg% at the end of pregnancy. In cases of fetal malformations, 68% of the glucose levels was below the tenth percentile of normal values. Hydramnios showed no deviation from normal values. In patients with abnormal glucose tolerance, AF glucose increased by a total of 42% and by 67% in fetal hyperinsulinism. Insulin-dependent diabetics had glucose values elevated by a total of 77% and by 106% in fetal hyperinsulinism. The AF glucose profile reflects the level of maternal blood glucose that is transported to the fetus and excreted in the fetal urine as a major source of glucose in AF.     

Disappearance of para-amino-hippurate from amniotic fluid.

Eight pregnant women and three pregnant sheep received 400 mg of para-amino-hippurate (PAH) intraaminotically. Serial samples of amniotic fluid and maternal blood were obtained. In sheep samples of fetal blood were also withdrawn. PAH appeared in maternal plasma in all the cases. In all pregnant women PAH disappeared slowly from amniotic fluid (50% in 4 hours). In one ewe the study was performed as in humans and showed the same pattern of disappearance. In the other two, fetal urine was drained outside the amniotic fluid and PAH disappeared from it at a much faster rate (90% in 4 hours). PAH concentration in fetal urine was 100 times higher than in fetal plasma. Our findings in pregnant women seem to suggest that PAH disappears from the amniotic sac by a diffusion mechanism. On the other hand the results found in sheep also suggest that the fetus may have an active role in PAH concentration in amniotic fluid, eliminating part of the substance into maternal blood across the placenta but returning a major portion to the amniotic fluid with fetal urine.     

Microvillar enzyme analysis in amniotic fluid and the prenatal diagnosis of cystic fibrosis

The activities of two microvillar enzymes, gamma-glutamyltranspeptidase (GGTP) and alkaline phosphatase (ALP) have been determined in amniotic fluid (AF) samples from 39 pregnancies with a 1-in-4 risk of cystic fibrosis. Seventeen of these were investigated prospectively. A reduced proportion of the fetal specific intestinal ALP isoenzyme was found in 7 of a total of 13 pregnancies with cystic fibrosis and in one pregnancy of confirmed normal outcome. Eight of the affected pregnancies were tested for AF GGTP activity and depressed levels were found in 15. None of the 3 liveborn cystic fibrosis cases in the prospective series was identified by the ALP assay although 2 had significantly reduced GGTP activity. There were several amniotic fluid samples from cases of cystic fibrosis, trisomy 18 and normal outcome which had discordant GGTP and ALP results. Four of the 6 cases of cystic fibrosis misclassified by the ALP assay had amniocentesis at 15 or 16 weeks gestation. Evidence is presented which confirms a previous suggestion that amniocentesis after 17 weeks gestation improves the predictability of the ALP isoenzyme assay for the prenatal diagnosis of cystic fibrosis.     

Fibrinolytic activity in amniotic fluid during late pregnancy

Fibrinolytic activity (FA) was assessed by the fibrin plate technique in 47 amniotic fluid samples obtained at 35 to 39 weeks' gestation. In amniotic fluid the FA (63.1 +/- 2.5 mm2) was considerably elevated when compared with previously reported serum levels of pregnant women. After centrifugation of the amniotic fluid samples a highly significant reduction of FA in the supernatant was recorded (42.9 +/- 1.3 mm2), suggesting that the cells of fetal origin are the main source of the high plasminogen activator activity demonstrated in amniotic fluid. A significant rise in amniotic fluid FA was demonstrated beyond the 37th gestational week. Furthermore, significantly greater FA was recorded in amniotic fluid samples from women with spontaneous onset of labor within a week. However, a single FA determination in amniotic fluid obtained by routine amniocentesis was found to be of little value for prediction of the time interval to delivery.     

Oxytocinase activity in human amniotic fluid and its relationship to gestational age

Oxytocinase (EC 3.4.11.3) activity in amniotic fluid samples obtained from 200 normal pregnant women (mean age, 28 years) between 14 and 40 weeks of gestation and during active labor was assayed using S-benzyl-L-cysteine-p-nitroanilide (BCN) and L-leucine-p-nitroanilide (LN) separately as substrates. With both substrates, amniotic fluid oxytocinase activity correlated inversely with gestational age; the level of the enzyme, which was highest in early pregnancy, decreased exponentially to a minimum near term and during labor. However, whether oxytocinase activity in the amniotic fluid is of any significance for the maintenance and/or termination of pregnancy remains to be established.     

Activated T cells in normal pregnant women and neonates

The phenotyping of T-cell subsets and T cells at different stages of activation was performed using a panel of monoclonal antibodies in samples from normal pregnant women at different stages of gestation and in the cord blood of neonates. The data obtained from pregnant women showed a slight decrease in the total number of T cells at the beginning of pregnancy, whereas there was a clear increase in 4F2-positive lymphocytes after a few months of gestation. No significant increase in Class II-positive lymphocytes was observed in normal pregnant women in comparison with adult healthy women. The data from neonates revealed a clear decrease of OKT3- and OKT4-positive cells and an increase of 4F2-positive cells in comparison with control subjects. These data indicate that alerted, but not fully activated, lymphocytes are present in the circulation of both the mother, after the first months of pregnancy, and the neonate. This finding reinforces the concept that during pregnancy there is an activation of certain immune components rather than a general depression of the immune system.     

Incidence of toxoplasmosis in 5532 pregnant women in Crete, Greece: management of 185 cases at risk

OBJECTIVES: To study the incidence of toxoplasmosis in pregnant women in Crete and to test a designed protocol for handling those at risk of delivering congenitally infected infants. STUDY DESIGN: Pregnant women were screened serologically over a period of 5 years. Cases with suspected acute toxoplasmosis were treated, peripheral blood (PB), and amniotic fluid (AF) tested by polymerase chain reaction (PCR) and culture, and fetuses monitored by ultrasonography. The absence of congenital infection in infants was confirmed by serology and clinical evaluation. RESULTS: Of the 5532 pregnant women followed, 70.57% remained seronegative, 29.45% were seropositive, and there was direct evidence of seroconversion in six cases. Acute toxoplasmosis was suspected in 185 cases, maternal parasitemia was detected in five cases and positive amniotic fluid in one case. Congenital infection was excluded in all infants followed, based on the absence of ultrasound findings in utero, lack of clinical symptoms at birth, negative Western blotting (WB) at birth and 3 months later, and descending serology for a year. CONCLUSION: Overall, 29.45% of the pregnant women followed were seropositive, 3.3% with suspected acute toxoplasmosis, and in 0.02% cases there was evidence of maternofetal transmission. The protocol tested allowed differentiation between acute and latent toxoplasmosis, safe management of the cases at risk and assisted in avoidance of unwarranted pregnancy terminations.     

Cortisol in amniotic fluid and cord blood in relation to prenatal betamethasone load and delivery.

The influence of maternal corticosteroid administration on the cortisol concentration in fetal blood and amniotic fluid (AF) was studied in women receiving betamethasone for prevention of IRDS. Thirty-four pregnant women in danger of spontaneous or induced preterm delivery were treated with 12 mg. of betamethasone daily for 3 days. AF was obtained by amniocentesis on the day before and the day following the betamethasone treatment and by amniotomy at delivery; cord arterial and venous blood was taken at delivery. Corresponding samples were obtained from 17 pregnant control subjects. All samples were analyzed in duplicate for cortisol by radioimmunoassay. The basal AF cortisol level rose with gestational age. The AF cortisol concentration fell from the basal value of 24.5 +/- 1.8 to 5.4 +/- 0.4 ng. per milliliter 3 days after the start of treatment, and it remained low at delivery if the treatment-delivery time was less than 1 week. An almost significant positive correlation (r = 0.543) was found between the cortisol concentration in cord arterial blood and AF. The cortisol concentration in cord blood in the controls was 108.2 +/- 14.3 ng. per milliliter in the arteries and 106.4 +/- 18.6 ng. per milliliter in the vein. Also, the cord blood cortisol level was depressed in betamethasone had been given within one week before delivery. The multifactorial influence on cord blood cortisol level prevents interpretation of the results as support for the concept that the human fetal adrenal is involved in labor initiation. The duration of gestation was not altered by the betamethasone treatment. The analysis of cortisol in the easily accessible amniotic fluid is suggested for estimating the function of the fetal adrenal cortex.     

Outcomes following intra-amniotic instillation with indigo carmine to diagnose prelabor rupture of membranes in singleton pregnancies: a single center experience

Objective: To evaluate clinical outcomes of women with singleton pregnancies that underwent intra-amniotic dye instillation (amniodye test) following equivocal diagnosis of prelabor rupture of membranes (PROM).

Method: Records of 34 pregnant women who underwent amniodye test for equivocal PROM were reviewed. Comparisons of characteristics, amniotic fluid (AF) cultures, AF interleukin (IL)-6 concentrations, and placenta pathology results between women who tested positive and those who tested negative were performed. A sub-analysis of women who were amniodye test-negative was also performed.

Results: (1) Commonest indication for amniodye test was a typical history of PROM with positive conventional tests and persistently normal AF volume, (2) amniodye test-positive women had a shorter procedure-to-delivery interval (p?=?0.008), and a greater proportion of histologic acute chorioamnionitis (p?=?0.04) and funisitis (p?=?0.01) than amniodye-negative women, and (3) in addition to similarities to women with amniodye-positive test, amniodye test-negative women who delivered <34 weeks, had a greater proportion of women with risk for preterm birth (p?=?0.04), than their counterparts who delivered between 34 0/7 and 36 6/7 weeks.

Conclusion: Equivocal diagnosis of PPROM should warrant an amniodye test to avoid iatrogenic intervention in women with intact amniotic membranes. AF analysis should be performed in amniodye test-negative women.