OBJECTIVETo detect the change of nerve growth factor (NGF) level in human amniotic fluid during gestation, and to explore the relationship between this change and fetal ventriculomegaly (VM).

METHODSThe studied subjects (collected from 2004 to 2007) were divided into four groups, including the second-trimester pregnancy group (n=113), third-trimester pregnancy group (n=110), fetal cerebral VM group (n=12), and healthy control group (n=12) which matched with the VM group in gestational weeks. The amniotic fluid specimens were obtained during amniocentesis or cesarean section. The NGF levels in amniotic fluid were detected with enzyme-linked immunosorbent assay.

RESULTSA significantly negative correlation was found between gestational age and the NGF level in amniotic fluid (r=−0.6149, P<0.0001). The NGF level in patients with fetal VM was significantly lower than that in healthy controls (33.95±29.24 pg/mL vs. 64.73±16.21 pg/mL, P=0.024).

CONCLUSIONNGF levels in amniotic fluid may be a sensitive marker for fetal VM.

Adult ; Amniotic Fluid ; chemistry ; Female ; Humans ; Hydrocephalus ; metabolism ; Nerve Growth Factor ; analysis ; Pregnancy

OBJECTIVETo investigate the feasibility of genetic diagnosis of Down's syndrome (DS) using short tandem repeat (STR), and to develop a rapid and accurate method for diagnosing DS.

METHODSQuantitative fluorescence polymerase chain reaction (QF-PCR) was used to amplify STR loci D21S11, D21S1440 and Penta D of 719 samples. Three hundred and eighty-nine samples were peripheral blood, 282 were amniotic fluid, 48 were chorionic villous samples. The products were analyzed using eleterophoresis to detect DS.

RESULTSAmong 652 samples with a normal karyotype, 635 showed 2 bands with a 1:1 ratio or a single band. The remaining 17 samples showed 3 bands, and were regarded as false positive results. For 67 DS samples, 53 showed 3 bands/peaks with a 1:1:1 ratio and 14 showed 2 bands/peaks with a 2:1 ratio. The sensitivity and specificity of STR loci D21S11, D21S1440 and Penta D were 76.12% and 98.62%, 71.64% and 98.93%, 89.55% and 99.85%, respectively. The overall sensitivity and specificity of 3 STR loci were 100% (67/67) and 97.39% (635/652), respectively.

CONCLUSIONCompared with conventional method, author's method is simpler, more stable and rapid, and can be used for large-scale prenatal screening of DS.

Amniotic Fluid ; chemistry ; Chorionic Villi ; chemistry ; Down Syndrome ; diagnosis ; genetics ; Female ; Humans ; Microsatellite Repeats ; Pregnancy ; Prenatal Diagnosis ; methods ; Sensitivity and Specificity

OBJECTIVETo investigate the application of fluorescence in situ hybridization (FISH) technique in prenatal diagnosis of complex chromosomal abnormalities.

METHODSEleven prenatal diagnosis cases (8 from amniocentesis and 3 from cord blood) with complex chromosomal abnormalities detected by routine G-banding, were further analyzed by FISH.

RESULTSThe FISH technique confirmed the results of balanced chromosome rearrangements detected by G-banding, and clarified the structure of the derivative chromosomes in the 3 amniocentesis samples and the origin of the mark chromosomes in the 2 cord blood samples.

CONCLUSIONFISH can be used to diagnose the complex chromosomal abnormalities accurately in prenatal diagnosis, and can provide very useful genetic information for clinical diagnosis and treatment.

Amniotic Fluid ; chemistry ; Chromosome Aberrations ; Female ; Fetal Blood ; chemistry ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Pregnancy ; genetics ; Prenatal Diagnosis ; methods

OBJECTIVETo study the effects of meconium-stained amniotic fluid on the cord blood IgE level in neonates.

METHODSA total of 404 neonates with meconium-stained amniotic fluid who were born by cesarean delivery between August 2003 and August 2005 (meconium-stained group) and 256 neonates with normal amniotic fluid delivered by cesarean (control group) were enrolled in this study. The meconium-stained group consisted of 80 cases of mild, 62 cases of moderate and 262 cases of severe meconium-stained amniotic fluid. The cord blood IgE level was measured using ELISA.

RESULTSThe cord blood IgE level in the meconium-stained group was statistically higher than that in the control group (t = 4.03, P < 0.01). There were significant differences between the mild and severe meconium-stained subgroups and the control group for the cord blood IgE level (F=4.28, P < 0.01). The cord blood IgE level in neonates with premature rupture of the membrane between the meconium-stained and the control groups was statistically different. Sexes, gestational age, birth weight and birth order were not associated with the IgE level of cord blood.

CONCLUSIONSThe cord blood IgE levels in neonates with meconium-stained amniotic fluid increase. Premature rupture of the membranes may be associated with an increase of cord blood IgE level.

Amniotic Fluid ; Asthma ; etiology ; Fetal Blood ; chemistry ; Humans ; Immunoglobulin E ; blood ; Infant, Newborn ; immunology ; Meconium ; Meconium Aspiration Syndrome

OBJECTIVES: To study the clinical features of severe meconium aspiration syndrome (MAS) and early predicting factors for the development of severe MAS in neonates with meconium-stained amniotic fluid (MSAF). METHODS: A total of 295 neonates who were hospitalized due to Ⅲ° MSAF from January 2018 to December 2019 were enrolled as subjects. The neonates were classified to a non-MAS group (n=199), a mild/moderate MAS group (n=77), and a severe MAS group (n=19). A retrospective analysis was performed for general clinical data, blood gas parameters, infection indicators, and perinatal clinical data of the mother. The respiratory support regimens after birth were compared among the three groups. The receiver operating characteristic (ROC) curve and multivariate logistic regression analysis were used to investigate predicting factors for the development of severe MAS in neonates with MSAF. RESULTS: Among the 295 neonates with MSAF, 32.5% (96/295) experienced MAS, among whom 20% (19/96) had severe MAS. Compared with the mild/moderate MAS group and the non-MAS group, the severe MAS group had a significantly lower 5-minute Apgar score (P<0.05) and a significantly higher blood lactate level in the umbilical artery (P<0.05). Compared with the non-MAS group, the severe MAS group had a significantly higher level of interleukin-6 (IL-6) in peripheral blood at 1 hour after birth (P<0.017). In the severe MAS group, 79% (15/19) of the neonates were born inactive, among whom 13 underwent meconium suctioning, and 100% of the neonates started to receive mechanical ventilation within 24 hours. Peripheral blood IL-6 >39.02 pg/mL and white blood cell count (WBC) >30.345×10⁹/L at 1 hour after birth were early predicting indicators for severe MAS in neonates with MSAF (P<0.05). CONCLUSIONS Meconium suctioning cannot completely prevent the onset of severe MAS in neonates with MSAF. The neonates with severe MAS may develop severe respiratory distress and require mechanical ventilation early after birth. Close monitoring of blood lactate in the umbilical artery and peripheral blood IL-6 and WBC at 1 hour after birth may help with early prediction of the development and severity of MAS.
Amniotic Fluid/chemistry* ; Female ; Humans ; Infant, Newborn ; Interleukin-6 ; Lactates ; Meconium ; Meconium Aspiration Syndrome/diagnosis* ; Pregnancy ; Retrospective Studies

OBJECTIVETo establish a prediction model of fetal meconium-stained amniotic fluid in re-pregnant women with intrahepatic cholestasis of pregnancy (ICP).

METHODSClinical data of 180 re-pregnant women with ICP delivering in Women's Hospital, Zhejiang University School of Medicine between January 2009 to August 2014 were collected. An artificial neural network model (ANN) for risk evaluation of fetal meconium-stained fluid was established and assessed.

RESULTSThe sensitivity, specificity and accuracy of ANN for predicting fetal meconium-stained fluid were 68.0%, 85.0% and 80.3%, respectively. The risk factors with effect weight >10% were pregnancy complications, serum cholyglycine level,maternal age.

CONCLUSIONThe established ANN model can be used for predicting fetal meconium-stained amniotic fluid in re-pregnant women with ICP.

Amniotic Fluid ; chemistry ; Cholestasis, Intrahepatic ; pathology ; Female ; Fetus ; Humans ; Infant, Newborn ; Meconium ; chemistry ; Neural Networks (Computer) ; Pregnancy ; Pregnancy Complications ; pathology ; Sensitivity and Specificity

OBJECTIVETo establish a multicolor primed in situ labeling (PRINS) protocol for chromosome detection in uncultured amniocytes.

METHODSChromosomes 18, X and Y in uncultured amniocytes were simultaneously detected by using the non-ddNTP-blocking multicolor PRINS procedure.

RESULTSWithin 7 h, the 3 chromosomes were simultaneously marked in the same uncultured amniocyte. The chromosome signals were successfully detected in 69 uncultured samples of amniotic fluid. The results were consistent with that obtained by chromosomes in cultured amniocytes.

CONCLUSIONThis multicolor protocol was high throughput, fast, simple, sensitive and reliable in diagnosing chromosome abnormalities in uncultured amniocytes.

Adolescent ; Adult ; Amniotic Fluid ; chemistry ; cytology ; Cells, Cultured ; Chromosomes, Human, Pair 18 ; chemistry ; genetics ; Chromosomes, Human, X ; chemistry ; genetics ; Chromosomes, Human, Y ; chemistry ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Pregnancy ; Prenatal Diagnosis ; methods ; Primed In Situ Labeling ; methods ; Young Adult

OBJECTIVETo investigate the application value of the multiplex ligation-dependent probe amplification (MLPA) technique in diagnosis and prenatal diagnosis of chromosomes 13, 18, 21, X and Y aneuploidy.

METHODSForty-four cases including 30 peripheral blood samples, 10 fetal cord blood samples, and 4 amniotic fluid samples were collected in this study. DNA was isolated from the samples and detected by MLPA, followed by analyzing in ABI310 Genetic Analyzer. Analysis of copy number changes for chromosomes 13, 18, 21, X and Y was carried out with RH-MLPA-analysis software. The routine karyotype analyses were also done for all the samples.

RESULTSOf 44 samples, the results of 42 by MLPA method was consistent with that by chromosome karyotyping. Only one case with trisomy 21 chimerism was failed to reach conclusion. In addition, one case of mark chromosome segment was identified as Y-chromosome segment by MLPA, while karyotyping failed to make judgment. The accurate rate of MLPA was 97.7% (43/44).

CONCLUSIONThe MLPA technique can simultaneously detect dozens of different target sequences and their copy number changes in a single reaction. It showed high specificity, good reproducibility, was fast and high-throughput. The MLPA technique can be applied to diagnosis and prenatal diagnosis of the common chromosomal aneuploidy.

Amniotic Fluid ; chemistry ; Aneuploidy ; Chromosomes, Human, Pair 13 ; DNA ; genetics ; isolation & purification ; DNA Copy Number Variations ; Down Syndrome ; diagnosis ; genetics ; Female ; Fetal Blood ; chemistry ; Humans ; Nucleic Acid Amplification Techniques ; methods ; Pregnancy ; Prenatal Diagnosis ; methods ; Sensitivity and Specificity

OBJECTIVETo explore the origin and mechanism of small supernumerary marker chromosomes (sSMC) in order to facilitate genetic counseling.

METHODSChromosome karyotypes of two fetuses and their immediate family members were analyzed by conventional G banding. High-throughput whole genome sequencing was used to determine the origin of sSMCs.

RESULTSFetus 1 was shown to have a karyotype of 47,XY,+mar but with normal FISH and B ultrasound findings. Its father also had a 47,XY,+mar karyotype with normal FISH results and clinical phenotype. High-throughput genome sequencing revealed that fetus 1 and its father were both 46,XY,dup(21)(q11.2;q21.1) with a 6.2 Mb duplication of the long arm of chromosome 21. The fetus was born with normal phenotype and developed well. Its grandmother also had a karyotype of 46,XX,t(15;21)(q13;p13) with normal FISH result and clinical phenotype. The karyotypes of its mother and grandfather were both normal. Analysis of fetus 2 showed a 47,XY,+mar karyotype with normal FISH results. High-throughput genome sequencing suggested a molecular karyotype of 46,XX. The fetus was born with normal phenotype and developed well. The karyotypes of its parents were both normal.

CONCLUSIONConsidering their variable origins, identification of sSMC should combine conventional G banding analyses with high-throughput whole genome sequencing for precise delineation of the chromosomes.

Adult ; Amniotic Fluid ; chemistry ; Chromosome Banding ; Chromosome Disorders ; diagnosis ; embryology ; genetics ; Cytogenetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genetic Markers ; Humans ; In Situ Hybridization, Fluorescence ; Infant, Newborn ; Karyotyping ; Male ; Pregnancy ; Prenatal Diagnosis ; Young Adult

To evaluate the implication of methymalonic acid (MMA) in the early diagnosis of neural tube defects (NTD), a quantitative assay for MMA was established by using gas chromatography-mass spectrometry with stable isotope of MMA as an internal standard. Amniotic fluid and maternal urine MMA concentration, maternal serum folate, red blood cell folate and vitamin B12 levels were measured in the middle term of NTD-affected and normal pregnancies. Amniotic fluid and maternal urine MMA concentrations in the middle term of NTD affected pregnancies (1.4 +/- 0.9 micromol/L, and 22.1 +/- 12.6 nmol/micromol creatinine) were significantly higher than that of normal pregnancies (1.0 +/- 0. 4 micromol/L, and 2.5 +/- 1.1 nmol/micromol creatinine). There was no significant difference between normal and NTD pregnancies for serum folate, red blood cell folate and vitamin B12 levels. The results suggested that MMAs in amniotic fluid and maternal urine are sensitive markers for early diagnosis of NTD. Vitamin B12 is an active cofactor involved in the remethylation of homocycteine and its deficiency is an independent risk factor for NTD. MMA is a specific and sensitive marker for intracellular vitamin B12 deficiency. This study suggests that it is necessary to monitor the vitamin B12 deficiency and advocates vitamin B12 supplementation with folate prevention program.
Adult ; Amniotic Fluid ; chemistry ; Biomarkers ; analysis ; urine ; Female ; Folic Acid ; blood ; Humans ; Methylmalonic Acid ; analysis ; urine ; Neural Tube Defects ; diagnosis ; metabolism ; Pregnancy ; Pregnancy Trimester, Second ; Prenatal Diagnosis ; Vitamin B 12 ; blood