Genetic analysis and prenatal diagnosis of a pedigree with oculo-facio-cardio-dental syndrome: a case report and literature review / 中华围产医学杂志

Objective:To analyze the clinical phenotypes and prenatal diagnosis of a pedigree with oculo-facio-cardio-dental (OFCD) syndrome.Methods:A pregnant woman at 17 gestational weeks was admitted to the Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School in 2017 for genetic counseling. Genetic tests as performed for the proband (the pregnant woman), her husband, and the induced fetus of last pregnancy genetic test and the detected variants were analyzed and verified by chromosomal microarray analysis (CMA), multiplex ligation-dependent probe amplification (MLPA) and quantitative real time-polymerase chain reaction (Q-PCR). The detection platform established by MLPA and Q-PCR technology was used to perform prenatal diagnosis of the present pregnancy. Other family members were screened for BCOR gene mutation. Related mutation types were retrieved from ClinVar database with term of " BCOR", and related literature from CNKI and PubMed with terms of "OFCD syndrome", " BCOR gene", and "oculo facio cardiac dental syndrome" to summarize the clinical manifestations, mutation type and pathogenesis of this disease. Results:The proband has congenital cataracts, long face, congenital atrial septal defect, and severe dental malformations, which were consistent with the clinical features of OFCD syndrome. WES suggested that the proband and her induced fetus were suspected of having a large submicroscopic deletion of the exons of BCOR gene, which was confirmed by CMA, MLPA and Q-PCR, with a 105 kb deletion containing BCOR exons 1-15. The amniotic fluid genetic analysis of the present pregnancy showed that the fetus has a normal female karyotype, and did not carry the same BCOR gene copy number abnormality as the proband. The child grew and normally developed without any characteristic manifestations of OFCD syndrome during follow-up. Other families of the proband did not show clinical features of OFCD syndrone, and no BCOR gene copy number abnormality was detected. A total of 35 cases of BCOR gene mutation types related to OFCD syndrome were retrieved from ClinVar database. The data analysis revealed that the differences in clinical manifestations between Lenz microphthalmos syndrome and OFCD syndrome were mainly caused by different mutation types of BCOR gene. Among the 90 retrieved cases of OFCD syndrome obtained through literature, only one case was reported in China. Analysis of these 90 cases showed that the characteristic manifestations of OFCD syndrome, involving the eye, face, heart, teeth, and skeletal system. OFCD syndrome were confirmed in the proband and her induced fetus according to the clinical manifestation and the mutation type of BCOR gene. Conclusions:The clinical manifestations of OFCD syndrome are complicated, caused by various mutation types of BCOR. Systematic molecular genetic technology can be effectively applied for gene and prenatal diagnosis of OFCD syndrome.

Analysis to the failure rate and causes of noninvasive prenatal testing based on high-throughput sequencing / 中华医学遗传学杂志

OBJECTIVE@#To analyze the cause and pregnancy outcome for non-reportable cell-free DNA (cfDNA) results during non-invasive prenatal testing (NIPT).@*METHODS@#cfDNA was extracted from maternal plasma from 5898 singleton pregnancies at 12 to 22 gestational weeks and underwent NIPT with strict quality control standards. For those with sub-standard results, redraw or invasive prenatal procedures were recommended.@*RESULTS@#Among the 5898 cases, 32 have failed for the initial NIPT, including 17 cases with substandard cffDNA%, 10 cases with data fluctuation after twice library constructing and sequencing, and 5 cases with unidentifiable sex chromosome abnormalities. For these 32 cases, 2 directly underwent amniocentesis, and karyotyping analysis showed both were normal. Six of the 30 redrawn cases finally turned out to be nonreportable. The final nonreportable rate was therefore 0.1% (8/5898). Of the redrawn cases, 1 trisomy 21, 1 trisomy 18 and 1 trisomy 13 high risk-cases were identified, which were all confirmed to be false positive. Among the 6 nonreportable cases, 2 women underwent invasive prenatal testing, and 1 was found to have a normal fetal karyotype, while another was found to have an abnormal karyotype of mos45,X[32]/46,XY[18]. The other 4 nonreportable cases who did not accept invasive prenatal testing have all reported normal child development at follow-up.@*CONCLUSION@#The main reason for nonreportable NIPT results was low cffDNA%. The high success rate of the redrawn cases has effectively increased the overall NIPT success rate and reduced the number of the cases necessitating invasive prenatal diagnosis. The initially nonreportable women may consider retesting after careful counseling with informed consent.

Health economic evaluation of five prenatal screening strategies for Down's syndrome / 中华围产医学杂志

ObjectiveTo investigate the cost-effectiveness and cost-benefit of five screening strategies for Down syndrome (DS) to optimize prenatal screening strategy.MethodsA retrospective analysis was conducted in 26803 gravidas, who underwent the second trimester maternal serum screening ( maternal serumα-fetoprotein andβ-human chorionic gonadotropin) from 2002 to 2003, whom were classified into three groups according to the results of serum DS screening: high risk group (≥1/270), borderline risk group (≥1/1000-<1/270) and elderly gravida group (age at expected date of confinement≥35 years old). TreeAge Pro 2011 sofware was used to set up the decision tree model for cost-effectiveness and cost-benefit analysis. Strategy 1: Maternal serum screening was carried out on all gravidas, and then prenatal diagnosis was performed for women in high risk group. Strategy 2: Non-invasive prenatal testing (NIPT) was carried out on all gravidas, and then prenatal diagnosis was offered for women with positive or suspected results. Strategy 3: NIPT was only carried out on gravidas of advanced maternal age and maternal serum screening was performed on the rest population. Gravidas with positive or suspected positive results in NIPT or classified into the high risk group underwent prenatal diagnosis. Strategy 4: Maternal serum screening was carried out on all gravidas. Those at high risk received prenatal diagnosis, while those at borderline risk underwent NIPT first and followed by prenatal diagnosis if positive or suspected positive NIPT results were identified. Strategy 5: Maternal serum screening was carried out on all gravidas. Those at high or borderline risk would undergo NIPT followed by prenatal diagnosis if they were positive or suspected positive for NIPT.Results(1) Among 26803 gravidas, 1244 were at high risk group (4.64％) with five having trisomy 21; 3925 were at bordelrine risk (14.64％) with four having trisomy 21; 300 women were of advanced age (1.12％) with one having trisomy 21. (2) Cost-effectiveness analysis: the incremental cost-effectiveness ratios of strategy 3 and 4 were negative and that of strategy 1 was 0 with a cost-effectiveness ratio of 15833764.53. The incremental cost-effectiveness ratio of strategy 2 was 49865746.10, which was far greater than that of strategy 5 (63049.56). The cost-effectiveness ratio of strategy 4 is 586703.63, which was less than those of strategy 1,2 and 3 but higher than that of strategy 5. The average cost-effectiveness ratio of strategy 5 was the lowest (508431.20) among these five strategies, which meant that for every diagnosis of DS, strategy 5 had the lowest cost (508400 yuan). (3) Cost-benefit analysis: The benefits of strategy 4 and 5 were greater than their costs. Strategy 5 had the highest benefit-cost ratio, followed by strategy 4, 2, 3 and 1. (4) When other factors remained unchanged and only the acceptance rate of prenatal diagnosis was adjusted from 50％ to 100％, strategy 1 had the least cost expectation, followed by strategy 3, 5, 4 and 2. When the cost of NIPT was below 82.4 yuan, the cost expectation of strategy 2 that performed on all gravidas was the lowest, while when it was between 82.4 and 1827.2 yuan, the screening cost of strategy 5 was the lowest.ConclusionsStrategy 5 has the best cost-effectiveness and cost-benefit. It would be the best screening strategy for DS, if the cost of NIPT is between 82.4-1827.2 yuan.

Prenatal diagnosis and follow-up of a case with Lowe syndrome caused by interstitial deletion of Xq25-26 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To report on a sporadic case of Lowe syndrome diagnosed prenatally with ultrasound examination and genetic testing.</p><p><b>METHODS</b>Detailed sonographic fetal screening was performed by an experienced sonographer at 32 weeks of gestation. Fetal cranial magnetic resonance imaging (MRI) was applied to detect potential brain abnormality. Chromosomal microarray analysis (CMA) was conducted on amniotic fluid sample from the fetus and peripheral blood sample from the mother.</p><p><b>RESULTS</b>Congenital cataract and enlarged posterior fossa were detected by fetal ultrasound screening. Fetal cranial MRI found hypoplasia of the gyrus. CMA revealed that the fetus has carried a 633 kb deletion at Xq25-26.1 which encompassed the OCRL gene. The mother was a carrier of the same deletion. Clinical examination after birth confirmed that the neonate was affected with Lowe syndrome in addition with an atrial septal defect.</p><p><b>CONCLUSION</b>Prenatal diagnosis of Lowe syndrome without a family history largely depends on fetal imaging. Should cataract be found by ultrasound screening, fetal MRI may be considered to rule out central nervous system anomalies. CMA assay should also be considered to facilitate the diagnosis.</p>

Chromosome microarray analysis of four fetuses with abnormal karyotypes / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To carry out chromosomal microarray analysis (CMA) on four fetuses with abnormal karyotypes.</p><p><b>METHODS</b>Amniotic fluid samples were obtained and subjected to routine G-banded karyotyping analysis. CMA was applied for cultured amniocytes to determine alterations of gene dosage and chromosomal breakpoints.</p><p><b>RESULTS</b>Abnormal karyotypes were found in the parents of 3 fetuses. Parental karyotypes of the remaining fetus were normal. Imbalance chromosome rearrangements were revealed by CMA in all 4 cases.</p><p><b>CONCLUSION</b>CMA is an effective tool for the evaluation of clinical significance and delineation of the breakpoints involved in complex chromosomal rearrangements.</p>

Phenotypic and genetic analysis of a child with blepharophimosis, ptosis, epicanthus inverses syndrome and tetralogy of Fallot / 中华医学遗传学杂志

OBJECTIVE To determine the genetic cause of a child with blepharophimosis, ptosis, and epicanthus inverses syndrome and tetralogy of Fallot, and to correlate the phenotype with the genotype. METHODS Routine G-banding has been previously performed on the patient and her parents. Chromosome microarray analysis (CMA) was performed for the three individuals and the fetus. RESULTS Chromosomal analysis has suggested normal karyotypes for the child and her parents. However, a de novo 8.9 Mb deletion on chromosome 3q22.1-q23 was detected by CMA. The deleted region has encompassed 74 genes including 41 disease-related genes, and this is also the most frequent region involved in interstitial 3q deletion. Patients with deletion of this region often have a common feature of dysplasia of eyelids, as well as a spectrum of other anomalies according to different breakpoints, including microcephaly, skeletal anomalies, congenital heart defects, cranial anomalies, intellectual disability and developmental delay. The patient's phenotype was in accordance with such spectrum. Her parents and sib did not show this variation by CMA. CONCLUSION The de novo interstitial deletion of 3q22.1-q23 probably underlies the main clinical manifestation in this child. CMA can provide more detailed information and allow further investigation of the genotype-phenotype correlation.

Application of chromosome microarray analysis for prenatal diagnosis of a fetus with partial duplication of 1p and uniparental disomy of chromosome 6 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the genetic cause for a fetus with structural anomaly, and to correlate the phenotype with the genotype.</p><p><b>METHODS</b>Amniotic fluid was obtained following the revelation of structural anomaly by ultrasonography. Cell culture and direct DNA extraction were performed in parallel. G-banded karyotyping analysis and chromosome microarray analysis (CMA) were subsequently carried out.</p><p><b>RESULTS</b>G-banded karyotyping has suggested the fetus to be a normal male. However, CMA analysis has revealed the presence of a mosaic 3.24 Mb duplication of 1p36.33p36.32 (24%) and uniparental disomy (UPD) of chromosome 6. The genetic diagnosis for the fetus was therefore 46,XY, arr 1p36.33 p36.32(849,466-4,090,472)×2-3, (6)×2 hmz. The anomaly can probably explain the ultrasound findings in the fetus.</p><p><b>CONCLUSION</b>Compared with conventional cytogenetic methods, CMA has greater resolution and throughput, and can serve as a more efficient platform for the detection of chromosomal microdeletion, microduplication, loss of heterozygosity and UPD.</p>

Application of different technologies for distinguishing true and pseudo mosaicisms during prenatal diagnosis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To use different technologies to distinguish true and pseudo mosaicisms among cultured amniocytes in order to attain more accurate diagnosis.</p><p><b>METHODS</b>With informed consent, 20 mL of amniotic fluid was obtained from pregnant women at between 18 to 24 gestational week. Each amniotic fluid sample was processed as two separate lines for the culturing, observation, harvesting and analysis. All procedures were conducted conforming to the Technology Standards of Cytogenetic Prenatal Diagnosis of Fetal Chromosome Abnormalities issued by the Ministry of Health in 2010. Umbilical cord blood, fluorescence in situ hybridization (FISH), single nucleotide polymorphism array (SNP-array) and flow cytometer were applied when necessary.</p><p><b>RESULTS</b>Among 3910 cases, 128(3.3%) were detected as mosaicisms. Further analysis with the above technologies has verified 6 cases as true mosaicisms and the remaining 120 as pseudomosaicisms. For one case detected by karyotype analysis as 47, XXY/46, XY, the ratio of different cell lines was confirmed by FISH as 1:2. Another case, detected by karyotype analysis as 47, XX,+mar/46, XX (1:1), was verified by SNP-array as 18p duplication. A suspected polyploidy mosaicism was rejected by flow cytometry and cord blood karyotyping.</p><p><b>CONCLUSION</b>Two separate cell cultures are important for distinguishing true and pseudo mosaicisms. Combined FISH, SNP-array and flow cytometry can attain more reliable and accurate diagnosis for mosaicisms.</p>

Protective effects of resveratrol on acute lung injury induced by lipopolysaccharide in mices / 中国生化药物杂志

Purpose To investigate the effect of resveratrol on acute lung injury induced by lipopolysaccharide (LPS) in mice.Methods Acute lung injury (ALI) was induced by LPS in mice. The changes of lung airway inspiratory resistance (Ri), expiratory resistance (Re), and dynamic lung compliance (Cdyn) were observed with pulmonary function test apparatus. Brochoalveolar lavage fluid (BALF) was collected, and the concentrations of interleukin 1β (IL-1β)、interleukin 6 (IL-6) and tumor necrosis factory α (TNF-α) were measured by ELISA. The ratio of wet to dry weight was calculated to assess lung edema. Pulmonary vascular permeability was examined with injection Evans blue to judge the destructive extent of alveolar epithelial cell and endothelial. Pathological section was made, and the histopathological change was observed with light microscope.Results Resveratrol can inhibit the elevation of Ri and Re, and the descent of Cdyn. Simultaneously, resveratrol reduced the concentration of IL-1β, IL-6 and TNF-α,as well as the wet to dry weight ratio and the pulmonary vascular permeability significantly. Furthermore, it also could attenuate the lung injury on histopathology.Conclusion The results show that pretreatment with resveratrol has a protective effect on ALI induced by LPS. The ultimate inhibiting and release of inflammatory factors were involved in the mechanism of the effects.

Detection rate of chromosomal abnormalities in women with different indications for invasive prenatal diagnosis and procedure-related complications / 中华围产医学杂志

Objective To discuss the detection rate of chromosomal abnormalities in women with different indications for invasive prenatal diagnosis(amniocentesis and eordocentesis), and the procedure-related complications. Metheds A retrospective analysis was conducted on 1264 women, who underwent invasive prenatal diagnosis (1082 amniocentesis and 182 eordocentesis), and the procedure-related complications were reviewed. Results The indications for invasive prenatal diagnosis in these 1264 women were: increased risk at prenatal screening (651, 51.5%), advanced maternal age (≥35) (318, 25.2%), abnormal foundings through uhrasonograph (136, 10.8%),history of adverse pregnancy (88, 6.9%), one or two abnormal serologic markers (52,4.1%), and chromosomal balance translocation carrier in either one of the couple(19, 1.5%). Thirty-seven cases were found to be chromosomal abnormalities with clinic significance and the indications for them were: ultrasonic abnormality (20/136, 14.7%); increased risk at prenatal screening (12/651, 1.8%); one or two abnormal serologic markers (1/52, 1.9%); history of adverse-pregnant (1/88, 1.1%)chromosomal balance translocation carrier in either one of the couple (3/19, 15.8%); advanced maternal age (0/318). Among the 1264 cases, 5 experienced spontaneous abortion and the procedure-related fetal loss rates were 0.28% for amniocentesis (3/1082) and 1.09% for cordocentesis (2/182), P=0. 154. The rate of complications after cordocentesis was significantly higher than amniocentesis (9.89 % vs 0.18 %, P= 0.0001). Conclusions Routine fetal karyotyping should be prompted after prenatal ultrasonographic abnormalities. However, invasive prenatal diagnosis due to advanced maternal age alone is controversial. Amniocentesis is the fist choice for invasive prenatal diagnosis.

Application of multiplex quantitative fluorescent PCR with non-polymorphic Iod in prenatal diagnosis / 中华妇产科杂志

Objective To explore the feasibility of application of multiplex quantitative fluorescent PCR with non-polymorphic loci in prenatal diagnosis of aneuploidies. Methods From Mar 2006 to Nov 2007, a total of 63 samples were collected from the Department of Obstetrics and Gynecology, Affiliated Drum Tower Hospital of Medical College, Nanjing University, including 54 villous samples obtained for karyotyping because of spontaneous abortion, six anmiotic fluid samples of second trimester and three umbilical cord blood samples of third trimester. Blood samples of 60 healthy adults were obtained at the same time as a control group, including 30 males and 30 females. Non-polymorphic QF-PCR was performed on both testing group and control group for the detection of aneuploidies. The Amelogenin gene (AMXY) was selected as an internal control, and dosage quotiety (DQ) of each locus was calculated according to the known formula, ff DQ was between O. 7 and 1.3, the sample was considered as normal If the figure turned out to be >1.3 or <0.7, a potential duplication or deletion of the corresponding gene or chromosome was indicated. If the results implied numerical abnormalities in more than one euchromusome, sex chromosome aneupioidies should be considered. Cell culture and karyotyping were carried out for every sample simultaneously. The results of non-polymorphic QF-PCR were checked with karyotypes. Results ( 1 ) In the control group, all female samples presented only an AMX peak for sex chromosome while all males showed AMX and AMY amplified peaks. The AMY/AMX ratios were between 0.7-1.3, and SD was between 0.05-0.12. (2) Among 19 QF-PCR abnormal cases, 13 cases were proved by karyotyping. Of the six cases which turned out to be conflicting, one case of trisemy 18 shown by karyotyping was not completely detected by QF-PCR, a locus on chromosome 18 implied trisomy, while another turned out to be normal(DQ=1.28). Four cases were detected by non-polymorphic QF-PCR as trisemies but showed normal female karyotype because of maternal contamination during cell culture. A karyotyping]y ' 46, XY' case did not present an AMY peak. Thirty-six out of 44 (82%) normal results implied by non-polymorphic QF-PCR were in accordance with cytogenetic analysis. Of the other eight cases, one case which failed cytogenetic analysis was detected by QF-PCR as normal Four cases showed multiploidy by karyotyping but normal in QF-PCR analysis, including three eases of 69, XXX, one case of 92, XXXX and one case of 45,XX,rob(13;21). The other two cases that showed normal male results turned out to be normal female karyotypes. Conclusions Prenatal aneuploidy detection by non-polymorphic QF-PCR is feasible in a clinical diagnostic setting. With the advantages of high throughput, rapidness and low cost, this method shows a good prospect in clinical application.