Altered IGF-I receptor expression in chorioamniotic cells in premature rupture of fetal membranes

OBJECTIVE: To investigate the presence and distribution of type I insulinlike growth factor receptor (IGF-IR) in the cells of the chorioamniotic membrane and to search for any alterations occurring in IGF-IR expression in premature rupture of membranes (PROM) patients. STUDY DESIGN: Fetal membranes collected at delivery from 42 pregnancies between 36 and 40 gestational weeks were included in the study. Seventeen of 42 cases had premature rupture of membranes, and 25 cases had intact membranes prior to delivery. Paraffin sections of thefetal membranes were stained with IGF-IR antibody by the streptavidin-biotin-immunoperoxidase method. The staining was scored and compared statistically between PROM and control cases. RESULTS: The fetal membranes of PROM cases had significantly reduced IGF-IR expression in chorionic trophoblastic cells when compared with the control group (P = .006, X2). CONCLUSION: Our immunohistochemical findings revealed that chanlges in IGF-IR levels in choriolzic amniotic cells may play a pathogenetic role in PROM cases, but the mechanism is speculative and needs further investigation.     

Programmed cell death (apoptosis) as a possible pathway to metalloproteinase activation and fetal membrane degradation in premature rupture of membranes

OBJECTIVE: Increased matrix metalloproteinase 2 expression and activity are associated with premature rupture of fetal membranes. A proapoptotic protein produced in response to deoxyribonucleic acid fragmentation, p53, can bind to the matrix metalloproteinase 2 gene promoter and cause increased gene expression. It promotes apoptosis by inducing the expression of the proapoptotic bax gene and inhibiting the antiapoptotic bcl-2 gene. This study was undertaken to investigate the expression pattern of apoptotic elements in pregnancy complications that may cause increased expression of the gene for matrix metalloproteinase 2. STUDY DESIGN: Amniochorial membranes were collected from the following groups of women: (1) women with premature rupture of fetal membranes, (2) women with preterm labor and intact membranes, and (3) women with term labor after vaginal delivery. Deoxyribonucleic acid fragmentation was tested with ligation-mediated polymerase chain reaction and the terminal deoxynucleotidyl transferase-mediated biotinylated deoxyribonucleoside triphosphate end-labeling assay. Matrix metalloproteinase 2, p53, bcl-2, and bax gene expression patterns were studied with quantitative competitive polymerase chain reaction. Statistical analysis was performed with the Tukey-Kramer multiple comparison test. RESULTS: Quantitative competitive polymerase chain reaction documented a 10-fold increase in the expression of the gene for matrix metalloproteinase 2 in premature rupture of fetal membranes with respect to term and preterm labor. This induction coincided with an increase in the expressions of the proapoptotic genes p53 and bax and a drop in the expression of the antiapoptotic gene bcl-2. Ligation-mediated polymerase chain reaction revealed deoxyribonucleic acid fragmentation in specimens from premature rupture of fetal membranes and not in those from preterm labor or labor at term. Histochemical analysis documented fragmented deoxyribonucleic acid in chorionic and amniotic cells. CONCLUSION: This study suggests that apoptosis is associated with premature rupture of fetal membranes. Deoxyribonucleic acid fragmentation, associated with elevations in the levels of the two proapoptotic gene products evaluated (p53 and bax ) and a drop in the level of the antiapoptotic bcl-2, was seen in premature rupture of the fetal membranes. Induction of matrix metalloproteinase 2 may be a function of p53 gene expression increase in premature rupture of fetal membranes.     

细胞凋亡与胎膜早破的相关性研究

目的:探讨细胞凋亡增加是否为胎膜早破的一个发病机理或危险因素。方法:随机采集临产前剖宫产分娩的初产妇肘静脉血、羊水及胎膜,其中胎膜早破者34例,正常完好胎膜17例。①用ELISA测定各组血清、羊水中TNF-α的浓度:②用TUNEL原位标记术精确测定各组胎膜细胞凋亡指数;③用免疫组化法检测各组胎膜组织中Bax、Bcl-2、Fas、Caspase-3表达情况。结果:①胎膜早破组血清、羊水中TNF-α浓度明显高于正常对照组水平(P<0.001);且血清、羊水中TNF-α浓度有相关性,r=0.386。胎膜感染组血清、羊水中TNF-α浓度较无感染组高(P<0.001)。②胎膜早破组胎膜感染发生率高于对照组(P<0.05)。③胎膜早破组细胞凋亡指数明显高于正常组(P<0.001);感染组细胞凋亡指数较无感染组高(P<0.001)。④胎膜早破组促凋亡蛋白Bax、Fas、Caspase-3阳性表达率高于对照组(P<0.05),但抑凋亡蛋白Bcl-2阳性表达率两组间无统计学差异(P>0.05);胎膜感染组Bax、Bcl-2、Caspase-3阳性表达率与无感染组比较无统计学差异(P>0.05),但感染组Fas阳性表达率高于无感染组(P<0.05)。结论:基因、环境因素相互作用下的细胞凋亡增加可能是胎膜早破的一个重要发病机制,可独立致病,亦或与其他相关危险因素(如感染)协同致病。     

Myofibroblast differentiation in the connective tissues of the amnion and chorion of term human fetal membranes-implications for fetal membrane rupture and labour

An area of the fetal membranes, within the rupture tear after spontaneous delivery at term, exhibits altered morphology compared to more distal sites. It is characterized by marked swelling of the amniotic and chorionic connective tissue layers, consistent with structural weakness, and a marked reduction of the thickness of both the cytotrophoblast and decidual layers. These features, albeit less extreme, have been identified in fetal membranes in the lower uterine pole in patients prior to labour. In this study of pre-labour, labour-affected and post-labour term fetal membranes, we report that these regions are associated with an alteration in the phenotype of the vimentin positive mesenchymal cell population of the chorionic connective tissue reticular layer, and are consistent with myofibroblastic differentiation, i.e. alpha-smooth muscle actin (alpha-sma) expression. In the reticular layer of the lower uterine pole biopsies in the labour-affected group the numbers and densities of alpha-sma immunoreactive positive cells were 17-fold (P=0.04) and 8.5-fold (P=0.02) higher than in mid-zone biopsies. After delivery, in rupture line biopsies the numbers and densities were 50-fold (P=0. 002) and 36-fold (P=0.003) higher compared to mid zone biopsies. The percentage of the vimentin positive population positive for alpha-sma was 2-5 per cent in mid-zone biopsies compared to 49 per cent (P=0.03) in the labour-affected 'cervical' biopsies and 69 per cent (P=0.05) in the rupture line biopsies. Within the tear sites, alpha-sma positive cells were also detected within the fibroblastic layer of the amniotic connective tissue. Although there was no significant difference between the numbers and density of alpha-sma cells in the reticular layers between mid and lower uterine pole biopsies in the pre-labour group, in a proportion of patients the biopsies were similar to labour-affected biopsies indicating that this alteration occurs prior to clinically apparent labour in these patients. The incidence of alpha-sma positive cells in the reticular layer correlated with morphological changes within the fetal membranes, for example thickness of reticular (r(2)=0.349, P=0.0006) and amniotic connective tissue layers (r(2)=0.389, P=0.0002). This suggests that cellular activities associated with myofibroblastic differentiation in the reticular layer of the chorion may be associated with the observed connective tissue changes, fetal membrane rupture and labour.     

Evaluation of neutrophile elastase and isoprostane 8epiPGF2alpha concentrations in maternal and umbilical cord blood serum and in amniotic fluid in pregnancies complicated by premature rupture of membranes

OBJECTIVES: To evaluate the total isoprostane 8-epi-PGF2alpha and neutrophil elastase (NE) concentrations in pregnancies complicated by premature rupture of membranes (PROM). MATERIAL AND METHODS: 128 pregnant women were divided into four groups: pregnancies complicated by PROM between 24.-36.(PPBP-N) and between 38 a 41 weeks of gestation (PPBP-D), uncomplicated pregnancies between 24-36 gestation weeks (K1) and pregnancies delivered by cesarean section (before uterine contractions had started) after 38 weeks (K2). The concentrations of NE and isoprostane 8-epi-PGF2alpha were measured in maternal serum, cord blood serum and in the amniotic fluid. RESULTS: The following study revealed higher concentrations of NE in maternal serum and in the amniotic fluid than in the umbilical cord blood in PROM cases, and lower amniotic fluid than maternal serum concentrations in the control groups. Also, the levels of isoprostane differentiated between compartments in particular groups. In both groups complicated with PROM, higher maternal serum and amniotic fluid NE concentrations than in controls were found. There were no differences in isoprostane levels between the groups. CONCLUSIONS: 1. Higher concentrations of NE in maternal blood serum and in the amniotic fluid than in the umbilical cord blood in PROM cases, as well as lower amniotic fluid than maternal serum concentrations in the controls, may be connected with pathogenesis of PROM. 2. Differentiated maternal serum, cord serum and amniotic fluid isoprostane concentrations may suggest various intensity of oxidative stress in particular compartments. 3. Lack of differences in maternal serum, cord serum and amniotic fluid isoprostane concentrations may suggest similar intensity of oxidative stress in cases with PROM and intact membranes.     

Correlation of placental inflammatory changes in clinical infection in premature rupture of membranes

Maternal and neonatal white blood cell counts, neonatal pharyngeal culture, and histologic examination of placenta were done in 55 term pregnancies with premature rupture of fetal membranes (PROM) and in 52 term pregnancies without PROM to study the correlation between placental inflammation and clinical outcome. The results were as follows: (1) A higher rate of chorionitis and chorionic plate inflammation exist in patients with PROM. The inflammation rate of chorionic plate was 90% in patients with labor lasting longer than 12 hours, and only 38% in patients with a duration less than 12 hours. The difference was highly significant (P less than 0.01). (2) The positive rate of neonatal pharyngeal culture was higher in the PROM group than that in the control group (P less than 0.01). (3) Inflammation of chorionic plate was seen in 6/6 of the cases with neonatal infection. The corresponding rate was only 38% in cases without neonatal infection (P less than 0.01). It is suggested that in patients with PROM, efforts should be made to shorten the time of labor and the neonates should be watched for possible infections in the presence of inflammation of the placenta, particularly the chorionic plate.     

The role of extracellular inducer of matrix metalloproteinases in premature rupture of membranes

Objective: To investigate the role of matrix metalloproteinases (MMP-2, MMP-9) and their inducer (CD147) in premature rupture of membranes (PROM) at term labor.

Methods: In a cross-sectional study, 24 women aged 19–39, with 37–40-week pregnancy, and no clinical and histological signs of chorioamnionitis, were divided into two groups with and without PROM. The histological and immunohistochemical study of the fetal membranes was performed with polyclonal rabbit antibodies to MMP-2/MMP-9 and monoclonal rabbit antibodies to CD147.

Results: The analysis of MMP revealed the increase of MMP-9 expression in the amniotic epithelium during premature membrane rupture both in rupture area, and beyond it, and increased MMR-2 expression in the mesodermal cells. We also found high level of CD147 in the amniotic epithelium in PROM group. The above-mentioned changes were found in all areas of fetal membranes, regardless of the rupture localization.

Conclusions: The study results demonstrate the increased expression of MMR-2 and MMR-9, which regulate the catabolism of fetal membrane extracellular matrix proteins, in amniotic membranes of women with PROM at term labor. The increased expression of CD147 may be one of the mechanisms triggering PROM in the absence of infection.     

Amniotic fluid endothelin levels and the incidence of premature rupture of membranes.

Objective. The purpose of this prospective study was to record Endothelin 1 (ET1) concentrations in the second trimester amniotic fluid and in women who develop premature rupture of membranes (PROM), preterm premature rupture of the membranes (PPROM) and in women with uneventful pregnancies. Method. Amniotic fluid was retrieved by amniocentesis from 125 women in the second trimester of pregnancy. The levels of Endothelin were measured by a sensitive and specific radioimmunoassay. Results. From the 125 women included in the study 20 developed PROM and preterm PROM (13 PPROM and 7 PROM). The ET1 concentration was significantly higher (P<0.001) in PROM and PPROM than in normal pregnancy (96.4 vs. 43 pg/ml). The sub-analysis of the two rupture of membranes groups found that the concentration of ET1 was higher in the PPROM than in PROM (118 vs. 72 pg/ml). Conclusion. The amniotic fluid concentration of ET1 is elevated by the second trimester in women who later develop preterm PROM or term PROM.     

Programmed cell death (apoptosis) in placentas from normal pregnancy and pregnancy complicated by term (t) and preterm (p) premature rupture of membranes (PROM)

Objective: The aim of this study was to determine the apoptotic index using three different apoptotic markers: terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL); M30 cytoDEATH antibody and fluorescein isothiocyanate (FTIC)-labeled annexin-V in the placenta and membranes from normal pregnancy and pregnancy complicated by premature rupture of membranes (PROM). Study design: Placentas from 16 pregnancies (22–40 weeks’ gestation) and 13 pregnancies complicated by pPROM and tPROM were collected at delivery. Maternal and gestational age, mode of delivery, gravidity and parity, fetal birthweight, Apgar scores, presence of histologic chorioamnionitis, interval between membrane rupture and delivery were recorded among PPROM and tPROM cases. Results: Only M30 cytoDEATH antibody (P=0.02) and TUNEL test (P=0.04) on fetal membranes gave statistically significantly higher levels in cases with premature rupture of membranes. The presence of histologic chorioamnionitis had no significant impact on apoptotic markers in PROM placentas. Preterm deliveries following the rupture of membranes had higher median AI values detected by M30_M antibody, compared to those cases delivered without PROM (P=0.03). Conclusion: High apoptotic nuclei counts were found in fetal membranes of pPROM.     

Caspase-8及Bcl-2表达与胎膜早破的相关性研究

目的:探讨Caspase-8及Bcl-2表达与胎膜早破的关系。方法:选择2003年6月~2004年5月我院足月妊娠自然分娩的孕妇48例,其中24例发生胎膜早破(胎膜早破组),24例未发生胎膜早破(对照组),病例均于阴道分娩后取胎膜破裂口处的胎膜组织5cm×5cm大,同时胎膜早破组在距胎膜破口处10cm以上的部位再取同样大的胎膜组织,胎膜组织均经石蜡包埋切片后采用免疫组化法测定Caspase-8及Bcl-2的表达。结果:(1)两组病例的胎膜组织均可见Caspase-8及Bcl-2的表达;(2)在胎膜早破组的胎膜组织中Caspase-8表达的阳性单位为6.89±0.19,对照组为2.33±0.06(P<0.01);而Bcl-2表达的阳性单位为9.55±0.24,对照组为21.37±0.32(P<0.01);(3)在胎膜早破组非破口部位胎膜组织中Caspase-8及Bcl-2表达的阳性单位分别为6.93±0.17和9·66±0.19,与破口部位胎膜组织中Caspase-8及Bcl-2表达的阳性单位比较无差异(P>0·05)。结论:胎膜早破的发生与Caspase-8的过度表达及Bcl-2表达的下调相关。     

Analysis of cervicovaginal fluid in the diagnosis of premature rupture of membranes

Premature rupture of membranes (PROM) complicates approximately 8% of all pregnancies, however in 3% of the cases the membrane rupture occurs before 37 weeks of gestation (preterm PROM--PPROM). The consequences of such an early loss of amniotic fluid are very serious. Serious complications and mortality of neonates born prematurely due to PPROM is very high and inversely proportional to gestational age. The diagnosis of PROM is based on medical history and physical examination. In case of doubt, a test determining the pH of vaginal secretions and ultrasonography with assessment of amniotic fluid index may be applied. In difficult cases vaginal fluid diagnostic markers are very helpful. Proposed markers include: beta-human chorionic gonadotropin (beta-HCG), placental alpha-microglobulin-1 (PAMG-1), insulin-like growth factor binding protein-1 (IGFBP-1) and alpha-fetoprotein (AFP). Other markers were also proposed, namely interleukin 6 (IL-6), prolactin, creatinine and lactates. Several tests for the analysis of cervico-vaginal secretions are already commercially available. In Poland various tests based on the analysis of vaginal pH and a test Actim Prom based on the analysis of IGFBP-1 are available. In clinical practice, approximately 10% of cases of PROM require additional confirmation, which can be difficult, especially in the outpatient setting. Various types of bedside tests, the so-called point-of-care testing--POCT are increasingly used in contemporary medical practice. Their implementation does not require a laboratory specialized equipment or trained personnel. This paper presents only those markers that either are already used in the diagnosis of PROM in the form of POCT testing, or could be relatively easily adapted for this purpose.     

解脲支原体宫内感染与胎膜早破关系的研究

目的 探讨解脲支原体(UU)宫内感染与胎膜早破的相关性。方法 运用培养法对妊娠晚期32例胎膜早破孕妇(试验组)和20例正常孕妇(对照组)的剖宫产术中羊水进行支原体培养检测,评价6种抗菌药物对抑制支原体的敏感性;同时运用PCR-微板核酸分子杂交法对上述羊水进行解脲支原体DNA检测。结果 试验组分离培养检测的UU感染阳性率为28.1％,对照组UU感染阳性率为5.0％(P>0.05);6种抗生素药敏试验提示氧氟沙星药物敏感性最弱;在同时进行的PCR-微板核酸分子杂交法试验中,试验组的UU检出率为46.9％,对照组的UU检出率为10.0％,两组比较差异有显著性(P<0.01)。结论 解脲支原体所致的宫内感染是部分妊娠晚期胎膜早破的原因之一。PCR-微板核酸分子杂交法联合培养法,是诊断解脲支原体宫内感染的特异、快速和准确的方法。     

Fetal activity in premature rupture of membranes

Fetal movement (FM) counts by mothers with premature rupture of membranes (PROM) may be helpful in estimating fetal well being, provided FM assessment is not altered by the reduced amniotic fluid. It is possible that the decreased uterine volume restricts fetal movements, though the closer contact of the fetus to the uterine wall may enhance maternal perception of even weak FM. The present study compared maternally perceived FM in 41 cases of PROM to 120 uncomplicated pregnancies. No significant differences in FM counts were detected between the two groups at any of the gestational ages. Fetal movement counts by mothers with PROM, therefore, can assist fetal surveillance as is the case in pregnancies with intact membranes.     

Amniotic fluid concentrations of adrenomedullin in preterm labor.

OBJECTIVE: To determine whether adrenomedullin levels in amniotic fluid were associated with preterm labor. METHODS: We measured immunoreactive adrenomedullin in amniotic fluid collected by amniocentesis from 36 women with clinical diagnosis of preterm labor or preterm premature rupture of membranes (PROM) and from 18 normal pregnant women. RESULTS: Amniotic fluid from cases of PROM and failure to respond to tocolysis were associated significantly with higher amniotic fluid adrenomedullin concentrations (177.0 +/- 22.5 pg/mL and 182.7 +/- 22.0 pg/mL, respectively, P < .01) than that from uncomplicated pregnancies (101.2 +/- 28.1 pg/mL) or preterm labor responsive to tocolysis (102.3 +/- 26.8 pg/mL). CONCLUSION: Amniotic fluid adrenomedullin is higher than normal in cases of PROM and preterm labor unresponsive to tocolysis, perhaps indicating enhanced synthesis from placenta or fetal membranes being stimulated by bacterial products.     

米非司酮药物流产后绒毛蜕膜细胞凋亡的相关研究

目的:探讨米非司酮对早孕绒毛蜕膜细胞凋亡、增殖的作用机制。方法:以20例药物流产患者为研究组,以20例非意愿妊娠要求行人工流产负压吸引刮宫术的患者为对照组,分别收集绒毛和蜕膜标本,应用原位末端标记法(TUNEL)检测细胞凋亡,并采用免疫组织化学方法检测bcl-2、bax、fas、fasL、增殖细胞核抗原(PCNA)5种蛋白在绒毛和蜕膜中的分布与表达强度,同时应用原位杂交法测定fas与fasLmRNA的分布与表达强度。结果:凋亡细胞在正常早孕绒毛合体滋养细胞中少量存在,蜕膜中偶见;绒毛、蜕膜中bcl-2、bax、fas、fasL、PCNA均有表达。采用米非司酮药物流产的绒毛合体滋养细胞及蜕膜间质及腺上皮细胞的凋亡显著增多,同时伴有促凋亡bax、fas、fasL蛋白及fasLmRNA含量的增加,而PCNA蛋白含量与C组比没有变化。结论:米非司酮不仅能促进早孕绒毛合体滋养细胞、蜕膜间质及腺上皮细胞的凋亡,而且主要通过Fas与FasL转录及翻译途径介导,bax表达增加也其也有一定的相关性,此可能为其抗早孕机制之一。     

Biomechanical analysis of human chorioamniotic membranes

The biomechanical properties of human fetal membranes were analyzed by means of a materials testing machine. Special attention was paid to the biomechanical properties of the intact chorioamniotic membrane and the amniotic and chorionic components, separately, and thickness and storage of membrane samples. The load-strain and stress-strain relationships, and parameters calculated from the curves: extensibility, strength, elastic stiffness and failure energy, express the visco-elastic behavior of these membranes. The mechanical properties of the chorioamniotic membranes are determined by the interaction between the amniotic and chorionic components of the membrane. The strength of the intact chorioamniotic membrane, however, is primarily determined by the amniotic component, because the amniotic component is much less extensible. Thus, the chorionic component only contributes 10-15% of the strength when the amniotic component breaks. The chorionic component is twice as extensible as the amniotic component. Samples of fetal membranes can be stored at -70 degrees C, with no significant changes in the biomechanical properties. No significant differences were found between specimens, which were oriented in parallel with and at right angles to the placental edge. Small samples can be analysed and the localization of samples in relation to the placental edge and rupture site of the membranes can be standardized. The method is well suited for studies of premature rupture of fetal membranes.     

Fetal growth restriction is associated with increased apoptosis in the chorionic trophoblast cells of human fetal membranes

The aim of the study was to determine the incidence and spatial distribution of apoptotic cell in fetal membranes obtained from human pregnancies complicated with fetal growth restriction (FGR) for which there was no established cause. Fetal membrane samples from normal (n=10) and FGR-affected (n=10) pregnancies were collected and stored following delivery. The incidence of apoptosis and the number of apoptotic cells in normal and FGR-affected fetal membranes were determined using immunohistochemistry and a monoclonal antibody for neo-epitope of cytokeratin-18, M30. The level of apoptotic proteins in FGR-fetal membranes compared to the normal tissue was determined using Western immunoblotting analysis. Multiple labeling of trophoblast cells using immunofluorescence markers was used to investigate regional differences in localization of apoptotic cells between normal and FGR-affected fetal membranes. Apoptosis was detected in both normal and FGR-affected fetal membranes. However, quantitative analysis of apoptotic cells by immunohistochemistry showed a significant increase in FGR-affected fetal membranes compared to normal (p<0.005). Furthermore, it was observed that apoptotic cells were predominantly localized to chorio-decidual layer of the fetal membrane. By using semi-quantitative analysis of Western immunoblotting, a significant increase in the levels of the apoptotic marker proteins poly-ribo (ADP) polymerase (PARP) and the neo-epitope of cytokeratin-18 were observed in FGR-affected fetal membranes compared to normal (p<0.005). Immunofluorescence studies further confirmed the restriction of the apoptotic cells to the chorionic trophoblast cells in FGR-affected fetal membranes. Our results document for the first time an increased incidence of apoptosis in FGR-affected fetal membranes, with the apoptotic cells restricted primarily to the chorionic trophoblast layer of the fetal membranes. Increased apoptosis in FGR-affected fetal membranes may impair functions of the fetal membranes that are important for normal fetal development and growth. Elucidation of the molecular mechanisms involved in the control of apoptosis in the chorionic trophoblast layer in the FGR-affected fetal membranes may provide further insights into the etiology of FGR.     

Amniotic fluid alpha 1-antitrypsin concentration in premature rupture of the membranes

Premature rupture of the membranes is probably a result of a loss in amniotic membrane collagen. A recent report that the concentration of alpha 1-antitrypsin was decreased in patients with premature rupture of the membranes suggested a generalized defect in such pregnancies. In this study we compared the concentration of alpha 1-antitrypsin in samples from pregnancies with premature rupture of the membranes and from pregnancies with preterm labor at similar gestational age. No difference in alpha 1-antitrypsin concentration was noted between these groups or between samples with or without intrauterine infection. These results support a localized inflammation and necrosis of the membranes at the site of rupture.     

Comparative study of three amniotic fluid markers in premature rupture of membranes: prolactin, beta subunit of human chorionic gonadotropin, and alpha-fetoprotein

OBJECTIVE: To evaluate whether prolactin, alpha-fetoprotein (AFP) or B-human chorionic gonadotropin (BHCG) is the most effective marker in vaginal fluid for diagnosing prelabor rupture of membranes (PROM). These proteins are present in amniotic and vaginal fluid and have been reported to be potent markers of PROM, but have not been used clinically nor compared to each other. STUDY DESIGN: A total of 100 pregnant women between 28 and 37 weeks of gestation were recruited for the study. Patients were divided into 2 groups. The first group consisted of 50 pregnant women diagnosed with ruptured membranes. The second group consisted of 50 normal pregnant women seen during routine antenatal clinic visit (control) group. All women underwent speculum examination aiming to sample prolactin, BHCG and AFP in the vaginal fluid. Ultrasonographic examination for gestational age and amniotic fluid index measurement was performed. The electrochemoluminescence (ECLIA) method was used for quantitative measurement of the three proteins (the total duration of the assay was 18 min). RESULTS: Vaginal fluid concentrations of the three markers were significantly higher in the PROM group than in the control group (p < 0.001). Receiver operator curve analysis indicated that AFP had 94% specificity, sensitivity, positive and negative predictive values, and efficiency. The other two markers have lower specificity, sensitivity, positive and negative predictive values, and efficiency: 70, 76, 71.7, 74.5 and 73% for prolactin and 72, 84, 75, 81.8 and 78% for BHCG, respectively. CONCLUSION: This work demonstrates that of the three markers investigated AFP has the highest diagnostic performance. Using the ECLIA method it can be an ideal marker for diagnosing PROM particularly in equivocal cases. The technique could be introduced into laboratory tests to meet clinical needs. Basel.     

米非司酮配伍米索前列醇抗早孕蜕膜与绒毛细胞凋亡及其调控基 …

目的 探讨细胞凋亡及其调控基因Ｂ－细胞淋巴瘤／白血病－２（ｂｃｌ－２）,ｂｃｌ－２相关Ｘ蛋白（ｂａｘ）基因的表达与药物终止早孕的关系,方法 对随机采用米非司酮负吸收（１５例米非司酮组）米非司酮并米索前列醇（米索）（１４例,米非司酮米索组）,负压吸收（１５例,对照组）终止早孕经碘化丙锭染色的蜕膜与绒毛组织,应用流式细胞定性定量分析技术进行ＤＮＡ分析,采用免疫荧光标记对Ｂｃｌ－２,ｂａｘ基因蛋白进行分