Serum prenatal screening in the second trimester is currently the prenatal screening method with the highest population coverage rate, and large-scale serum prenatal screening has become a trend. The Medical Genetics Committee of the Chinese Eugenics Science Association organized some experts in prenatal screening in China to jointly draft the " Expert Consensus on the Scientific Management of Serum Prenatal Screening" , which focusing on the organization and management of large-scale serum prenatal screening and quality control in the laboratory, for reference by domestic counterparts.

Objective To analyze the mutations of IDS gene in a mucopolysaccharidosis type Ⅱ (MPS Ⅱ) family and to make prenatal diagnosis on the high-risk fetus which has been pregnant for eleven weeks.Methods IDS gene was analyzed by bidirectional DNA sequencing in 2 patients and their mother,and 5 unaffected individuals.Prenatal diagnosis for the high-risk fetus was performed by chorionic villus sampling after the genotypes was identified.Results The mutation c.344delA (N115fsX15) was detected in the two patients,and the mother of patients carried the heterozygous c.344delA (N115fsX15) mutation.None of the mutant was detected in the 5 unaffected subjects.The fetus carried c.344delA (N115fsX15) heterozygous mutation and was a carrier.Conclusion The deletion mutation c.344delA (N115fsX15) is causative to the pedigree of MPS Ⅱ,and prenatal diagnosis is the efficient method to avoid defect birth.

Three-parent test tube baby technology is important to solve the mitochondrial genetic disease.Once available,it raises greatly ethical controversy such as breaking traditional family values,hitting the religious belief,existing unknown risks,correctly handling the failed embryo,as well as the influence on the social status of the babies.Regarding these controversy,we can discuss it from several aspects.Because the development of ethics is behind the progress of science and technology,we should affirm the value of three-parent test tube baby technology and balance the development of science and technology with respecting the religious beliefs.Strict supervision system and standard application system reflect our respect for life.Incomprehension to the unknown things should become the motivation of our inquiry.We should face up to our fear of three-parent test tube baby technology,and thus to strengthen research and deepen understanding.Based on the above argument,this paper puts forward the ethical principles that should be followed in the development of three-parent test tube baby technology,namely respect,benefit,no harm and justice.

Objective:To observe and analyze the pathogenic gene types and clinical phenotypes of Leber congenital amaurosis (LCA).Methods:A retrospective clinical study. Six patients with LCA confirmed by genetic testing and 18 family members were included in the study. The patients came from six unrelated families. The family was investigated with a specific hereditary eye disease enrichment panel which contained 463 known pathogenic genes and based on targeted exome capture technology first to indentify the potential pathogenic genes and mutations. Then the TULP1 , RPGRIP1 , GUCY2D pathogenic mutations were conformed by Sanger sequencing. The pathogenicity of the gene variation was searched through relevant databases and PubMed literature, and its function was explained by protein prediction software. Results:Of the 6 patients, 3 were males and 3 were females; the age was from 3 to 33 years. Nystagmus, finger pressing eyes, photophobia, and night blindness were seen in 5 cases; electroretinogram showed 3 cases of extinction or near extinction; and 4 cases of retinopathy. The results showed patients with compound heterozygous mutation of c.1318C> T and c.1142T> G, homozygous mutation ofc.1318C> T and compound heterozygous mutation of c.1153G> A and c.1561C> T of TULP1 in Family 1, Family 2 and Family 5, respectively. There were compound heterozygous mutations of RPGRIP1 c.391delG and c.1468-2A> G in Family 3 and c.715delA and c.1765C> T in Family 6, respectively. Homozygous mutation of c.3177_3178delAC of GUCY2D was found in Family 4.The parents of all six patients were carriers of corresponding heterozygous mutations. TULP1 gene c.1142T> G, RPGRIP1 gene c.391delG, c.715delA and c.1765C> T and GUCY2D gene c.3177_3178delAC mutations were novel mutations and unreported. The 381th amino acid locus of product protein of TULP1 gene was highly conserved among species. The protein prediction software predicted that the mutation pathogenic. The c.391delG, c.715delA and c.1765C> T mutations of RPGRIP1 gene and c.3177_3178delAC mutation of GUCY2D gene can lead to early translation termination of their product proteins, which are pathogenic variants. Conclusion:The pathogenic mutations of TULP1, RPGRIP1 and GUCY2D genes led to LCA 15, LCA 6 and LCA 1 in six families.

Objective:To analyze the genetic test results and ultrasonographic markers of 41 fetuses with short femurs and their relationship.Methods:This study retrospectively analyzed 41 fetuses who were diagnosed with short femurs by ultrasound during 19-37 gestational weeks and underwent prenatal genetic examination at the First Affiliated Hospital of Zhengzhou University from January 2018 to June 2019. According to the results of genetic examination, these cases were divided into three groups after excluding three cases of variants of unknown significance: genetically normal group, chromosome variation (including chromosomal aneuploidy and pathogenic or likely pathogenic copy number variations) group, and gene mutation (including pathogenic or likely pathogenic gene mutations) group. According to the head circumference (HC), abdominal circumference (AC) and femur length (FL), Z FL, FL/HC, FL/AC, ΔZ H-F and ΔZ H+A-2F for each fetus were calculated. One-way ANOVA and LSD- t test were used for statistical analysis. Results:(1) Among the 41 fetuses with short femurs, there were 28 in the genetically normal group, five in the chromosome variation group, three with chromosome variations of unknown significance and five in the gene mutation group. (2) In the genetically normal, chromosome variation and gene mutation groups, Z FL values were -2.78±0.77, -4.36±0.69 and -4.69±0.70; FL/HC ratios were 0.178±0.011, 0.170±0.010 and 0.131±0.022; FL/AC ratios were 0.197±0.013, 0.186±0.011 and 0.151±0.017; ΔZ H-F values were 2.49±1.09, 3.53±1.28 and 8.17±1.30; ΔZ H+A-2F values were 4.44±2.00, 6.78±2.20 and 14.28±1.26, respectively. The differences in Z FL values between the genetically normal group and the chromosome variation group as well as the gene mutation group were statistically significant (both P<0.05); so were the differences in FL/HC, FL/AC and ΔZ H-F values between the gene mutation group and the genetically normal group as well as the chromosome variation group (all P<0.05) and in any pairwise comparison of ΔZ H+A-2F among the three groups (all P<0.05). Conclusions:The genetic etiology of fetal short femurs is mainly related to chromosomal variations (including chromosomal aneuploidy and pathogenic or likely pathogenic copy number variations) and gene mutation. In fetuses with chromosome variation and gene mutation, the degree of the femoral development delay relative to the development of HC and AC is worse than that in the normal genetic results group.

Objective To identify the mutation of trafficking protein particle complex 2 (TRAPPC2) gene in a large Chinese pedigree with X-linked spondyloepiphyseal dysplasia tarda by the PCR-based capillary electrophoresis methods.Methods The blood samples were collected from a large Chinese pedigree of three generations with six affected persons with X-SEDT.Four exons comprising the TRAPPC2 gene open reading frame as well as their exor/intron boundaries were analyzed by argrose electrophoresis and bidirectional direct sequencing of PCR products.Fluorescence labeled fragment analysis was performed by capillary electrophoresis.Results A 5-bp deletion mutation of TRAPPC2 gene in exon 5,c.262_266delGACAT (D88del; I89fX12),was identified in the proband and his unaffected mother(a heterozygote) in the Chinese family with X-SEDT,but no other sequence change occurring in exons 3,4 and 6 was detected.The old sister of proband was determined being carriers because she carries the deletion fragment allele of exon 5 PCR product and the young sister being normal individuals because she carries the wild allele of TRAPPC2 gene.Conclusions The mutation c.262_266delGACAT (D88del; I89fX12) of TRAPPC2 gene was firstly reported in Chinese people.The mutation of c.262_266delGACAT (D88del; I89fX12) in TRAPPC2 gene may be the pathologic cause of the patients in the X-SEDT pedigree.Fragment analysis combined with DNA sequencing by capillary electrophoresis method is effective laboratory test in the small deletion mutation analysis and carriers screening in X-SEDT family.

Objective To analyse MMACHC mutations for 45 pedigrees with combined methylmalonic aciduria and homocyctinuria by Sanger sequencing, and to discuss the utility of prenatal genetic diagnosis for these pedigrees.Method Peripheral blood was collected from 45 probands and their parents from 2012-2015 in Genetic Counselling Clinic of the First Affiliated Hospital of Zhengzhou University, and the DNA were extracted from the blood.Then the coding sequence of MMACHC gene was amplified by PCR, and the PCR products were further sequenced to detect mutations for each pedigree.For 12 families, chorionic villus sampling was performed on the pregnant women to make prenatal genetic diagnosis.Result There were 14 distinct mutations detected in the 45 pedigrees, and the most frequent mutations are c.609G>A(W203X),c.658-660delAAG(K220del)and c.80A>G (Q27A).Two of those mutations have not been reported before:one is a splicing site mutation c.81+1G>A;while the other is a missense mutation c.665A>G,p.Y222C.Most mutations were found in exon 4.Among the 12 pedigrees who received prenatal diagnosis, 2 fetuses were normal, 7 fetuses were carriers of heterozygous mutation, and the other 3 fetuses were patients with compound heterozygous mutation or homozygous mutation.The couples whose fetuses were normal or carriers continued the gestation, while the couples whose fetuses were patients decided to terminate the pregnancy.After delivery, the outcome of the fetuses was the same as the prenatal diagnose results.Conclusion Two novel mutations of MMACHC were identified and prenatal genetic diagnosis helps to avoid the delivery of combined methylmalonic aciduria and homocyctinuria patients.

Objective To analyze the mutation of MUT with Sanger sequencing technology to explore the feasibility of its application in prenatal diagnosis.Methods MUT sequencing was performed in 24 pedigrees who had history of isolated methylmalonic acidemia (MMA) babies and came to the First Affiliated Hospital of Zhengzhou University and Newborn Screening Center of Maternal and Child Health Hospital of He'nan Province between October 2012 and June 2015 for genetic counseling.Meanwhile,another 100 cases of normal controls also had their MUT gene sequence analyzed.After confirming the genotype of each pedigree,we collected the villi of nine high-risk fetuses in nine pedigrees whose parents were prepared for prenatal diagnosis.Results Totally,25 kinds of MUT gene mutations were identified among the 24 isolated MMA pedigrees,in which 11 were novel mutations including one nonsense mutation [c.616C＞T(p.Q206X)],six missense mutations [c.613G＞A(p.E205K),c.894T＞G(p.1298N),c.1009T＞C(p.F337L),c.1154G＞T(p.L385W),c.1663G＞A(p.A555T) and c.1675G＞A(p.R559G) and four frame shift mutations [c.626-627insC(p.P209Pfs*2),c.755-756insA(p.H252Qfs*6),c.756-757insA(p.M253Nfs*5) and c.1581-1582insA(p.A528Ifs*4)].None of the above mutations was detected in the controls.Finally,among the nine pedigrees for prenatal diagnosis,two were determined to have normal MUT gene,four were found to be heterozygous mutation carriers of MUT gene and three were confirmed as complex heterozygous or homozygous mutation carriers.Families of fetus who had normal MUT gene or fetuses who were carriers chose to continue the pregnancy,while those who had heterozygous mutation of MUT gene chose termination.The results of follow-up of newborns were consistent with that of prenatal diagnosis.Conclusions We found two novel mutations in MUT gene that might lead to isolated MMA.And Sanger sequencing technology for MUT gene sequencing analysis might effectively avoid the birth of isolated MMA children.

Objective To compare the diagnostic significance of pleural SLPI,IFN-γ and ADA for differenti-ating TPE from pleural effusions with the other etiologies. Methods Pleural effusion samples were obtained from 93 patients who were divided into the following groups: tuberculous pleural effusion,malignant pleural effusion, bacterial pleural effusion and transudative pleural effusion. The pleural effusion and/or serum levels of SLPI , IFN-γand ADA were determined. Results 1.The concentrations of SLPI, IFN-γand ADA in tuberculous pleural effusion was higher than that in malignant group, bacterial group and transudative group. 2. The diagnostic value of SLPI, IFN-γor ADA for the diagnosis of tuberculous PE is high respectively. The combinations of SLPI, IFN-γand/or ADA gained the more valuable diagnostic performance. Conclusion Pleural SLPI, IFN-γand ADA may be helpful for the differential diagnosis of tuberculous pleural effusion and the other pleural effusion. The combinations of SLPI or/and IFN-γor/and ADA further increased diagnostic value.

Objective To analyze the variations of PTPS gene in patients with suspected 6-pyruvoyl-tetra hydropterin synthase deficiency (PTPSD) and to make prenatal diagnosis in high-risk families. Methods Chemiluminescence was used for phenylalanine detection in blood or dried blood spots.Patients with phenylalanine concentration over 120μmol/L were detected by urine pterin analysis, and the activity of dihydropteridine reductase (DHPR) was detected. tetrahydrobiopterin loading tests were performed in suspected patients with abnormal urinary pterin profiles. PTPS gene variation analysis was performed by direct Sanger sequencing based on PCR amplification. Prenatal diagnosis in 7 high-risk families was performed by chorionic villus sampling when the genotype was identified. Results In 656 patients with hyperphenylalanine, 22 cases were diagnosed as PTPSD clinically. 16 variations were detected in the 22 PTPSD cases. The 5 variations, p.Lys77Arg, p.Ile84Phe, c.315-2A>G, c.244-2A>T, c.187-1G>T, were identified as novel variations. Two fetuses carried the same mutation with the proband and therefore were thought to be PTPSD fetuses. Three fetuses carried only one mutant allele and thus were thought to be PTPSD carriers.