The effectiveness of expanded carrier screening based on next-generation sequencing for severe monogenic genetic diseases.

Expanded carrier screening (ECS) based on next-generation sequencing has been the subject of few studies to estimate the effectiveness of ECS in the Chinese population. A total of 3737 individuals from Southwest China or the general Chinese population, including 1048 pairs and 1641 individuals, were analysed by ECS for 155 monogenetic diseases. An ECS panel was used to detect 147 genes and 10,449 variants in 145 autosomal recessive and 10 X-linked recessive disorders. A total of 43.27% (1617/3737) were found to be carriers of at least one of the 155 monogenetic diseases. The average number of carriers of these recessive mutations was 0.54 and ranged from 0 to 4. Of the 1048 couples, 74.81% (n = 784) were found to have at least one partner carrying more than one disease. In addition, 5.34% of the couples at risk (n = 56) were heterozygous for the same autosomal recessive disease, and 0.37% of the women (9/2440) were carriers of X-linked diseases. Our study demonstrated the clinical significance of ECS in Chinese populations and the need for a programme of familial screening for the prevention of severe recessive monogenetic diseases.

[Prenatal diagnosis and genetic analysis of a fetus with der(X)t(X;Y)(p22.3;q11.2)].

OBJECTIVE: To explore the pathogenesis and genetic characteristics of a fetus with a der(X)t(X;Y)(p22.3;q11.2) karyotype. METHODS: G-banding karyotyping analysis, BoBs (BACs-on-Beads) assay, and single nucleotide polymorphism array (SNP-array) were used to delineate the structural chromosomal aberration of the fetus. The parents of the fetus were also subjected to karyotyping analysis. RESULTS: The fetus and its mother were both found to have a karyotype of 46,X,add(X)(p22), while the father was normal. BoBs assay indicated that there was a lack of Xp22 but a gain of Yq11 signal. SNP-array confirmed that the fetus and its mother both had a 7.13 Mb deletion at Xp22.33p22.31 (608 021-7 736 547) and gain of a 12.52 Mb fragment at Yq11.221q11.23 (16 271 151-28 788 643). CONCLUSION: The fetus was determined to have a karyotype of 46,X,der(X)t(X;Y)(p22.3;q11.2)mat. The combined use of various methods has facilitated delineation of the fetal chromosomal aberration and prediction of the risk prediction for subsequent pregnancy.

Growth retardation and congenital heart disease in a boy with a ring chromosome 6 of maternal origin.

BACKGROUND: Rare chromosomal structural abnormalities, including ring chromosomes, often pose challenges to clinical genetic counselling. RESULTS: Here, we report a newborn with congenital heart disease and developmental delay who inherited ring chromosome 6 [46,XY,r(6)(p25q27)mat] from a phenotypically normal mother. Genotypes and phenotypes were analysed by molecular cytogenetic analysis, whole-exome sequencing and literature review. CONCLUSIONS: Our study showed that the pathogenicity of the ring chromosome abnormality [r(6)(p25q27)] was mainly affected by chromosome imbalance, deletions of genes with haploinsufficiency, duplications of genes with triple sensitivity, parental inheritance of the imbalance and the imprinting status of the affected genes.

Ampelopsin-sodium induces apoptosis in human lung adenocarcinoma cell lines by promoting tubulin polymerization in vitro.

Previous studies have demonstrated that ampelopsin (AMP), a type of flavonoid isolated from the stems and leaves of Ampelopsis grossedentata, exhibits anti-cancer activity in various types of cancer. Conversion of AMP into its sodium salt (AMP-Na) conferred enhanced solubility and stability to it. The present study aimed to evaluate the anti-cancer activity of AMP-Na in human lung adenocarcinoma cell lines and to investigate its mechanisms of action. Cell proliferation and viability were assessed by MTT and colony formation assays, and cell migration was determined using a scratch wound healing assay. The cell cycle distribution, apoptosis rate and tubulin immunofluorescence intensity were analyzed using flow cytometry, the cell ultra-microstructure was examined using transmission electron microscopy and the accumulation of tubulin was determined using laser confocal microscopy. The results demonstrated that AMP-Na significantly inhibited the proliferation, clonogenicity and migration of human lung adenocarcinoma cells. Furthermore, AMP-Na induced SPC-A-1 cell apoptosis, and promoted tubulin polymerization. The results suggested that the underlying mechanisms of AMP-Na may involve targeting of microtubules and tubulin polymerization to subsequently disrupt mitosis and induce cell cycle arrest at the S-phase.

Possibility of early diagnosis in a fetus affected by Prader­Willi syndrome with maternal hetero­UPD15: A lesson to be learned.

Prader­Willi syndrome (PWS), a complicated neurodevelopmental disorder arising from errors in genomic imprinting, is characterized by evident hypotonia along with feeding difficulties and the absence of crying in early infancy. Hyperphagia and obesity are not uncommon in patients with PWS, usually accompanied by intellectual disability, cognitive impairment, short stature, small hands and feet, as well as hypogonadism and typical facial features. Due to the severe complications associated with PWS, a thorough understanding of its features and an early diagnosis, preferably in the fetal period, are important for clinical management. According to previous studies, prenatal diagnosis has been confirmed in only a few cases of PWS, using ultrasound, or as an accidental finding by cytogenetic molecular techniques, as no precise fetal phenotype has been defined. In this present study, an infant with PWS arising from maternal heterodisomy of chromosome 15 is described. This is a typical case of missed diagnosis by fetal ultrasound examination, chromosome karyotype analysis and chromosome microarray (CMA) conducted during the pregnancy. To delineate the complex prenatal characteristics of a fetus with PWS, prenatally­diagnosed cases of PWS described in the literature were reviewed. This present study indicated that although prenatal signs are not sufficient for a diagnosis to be confirmed, a comprehensive consideration of these signs is important in leading to a diagnosis of suspected PWS, and thus prompts further prenatal investigations using molecular genetic tools. Furthermore, this present study also suggested that CMA can lead to a missed diagnosis of PWS/Angelman syndrome and other imprinting disorders despite its high value in the detection of copy­number variants in individuals with developmental delay. If clinical signs strongly suggest PWS, other prenatal molecular genetic investigations, including methylation tests and short tandem repeat­based linkage analysis for uniparental disomy, are recommended as an additional tool to aid diagnosis.

[Association between Val80 polymorphism of the CYP19 gene, A163G polymorphism of the OPG gene and bone mineral density in post-menopausal Chinese women].

We investigated the association of the G/A polymorphism at Val80 of the cytochrome P450 family 19 (CYP19) gene, the A163G polymorphism of the osteoprotegerin (OPG) gene with bone mineral density (BMD) in 200 randomly selected postmenopausal women in Chongqing. Single nucleotide polymorphisms were detected by polymerase chain reac-tion-restriction fragment length polymorphism (PCR-RFLP). BMD of the proximal femur and lumbar spine (L2-4) was measured by NORLAND XR-46 dual-energy X-ray absorptiometer (DEXA). The frequencies of genotypes in these women were as follows: GG (19.5), GA (44.5%), AA (36.0%) for the Val80 polymorphism in CYP19; and AA (13.0%), AG (42.0%), GG (45.0%) for the A163G polymorphism in OPG. The distribution of genotype frequency was in Hardy-Weinberg equilibrium (P0.05). ANCOVA and multiple stepwise regression analysis showed the Val80 polymor-phism in the third exon of the CYP19 gene was not associated with the BMD in postmenopausal women (P0.05). Except for the trochanter region, BMD at the femoral neck, Ward's triangle, and L2-4 was lower in subjects with AG/GG/AG+GG genotypes than those with the AA genotype for the A163G polymorphism and A163G genotypes were associated with BMD at these skeletal regions in postmenopausal women (P0.05). A163G polymorphism resides in the promoter region of the OPG gene and its genotype distribution is significantly different among different ethnic groups. Our results indicate that the AA genotype might have some beneficial effect on BMD and the variant G allele might reduce BMD in postmenopausal women.

A rapid reversed phase high-performance liquid chromatographic method for determination of sophoridine in rat plasma and its application to pharmacokinetics studies.

A high-performance liquid chromatography (HPLC) method for determining sophoridine in rat plasma was developed for application in the pharmacokinetic studies. The plasma was deproteinized with acetonitrile that contained an internal standard (ephedrine) and was separated from the aqueous layer by adding sodium chloride and sodium carbonate. The HPLC assay was carried out using a YMC-ODS column. The mobile phase was methanol-ethanol-0.01 moll(-1) ammonium acetate buffer-triethylamine (10:0.5:89.5:0.03, v/v/v/v) (pH 6.80). The flow rate was 0.8 ml min(-1). The detection wavelength was set at 210 nm. The method was used to determine the concentration-time profiles of sophoridine in the plasma following oral administration or injection of sophoridine aqueous solution. The fractions of sophoridine reaching the systemic circulation were estimated for the first time by a deconvolution method.

Changes of soluble fas and soluble fas ligand in serum and peritoneal fluid of infertile patients with endometriosis.

OBJECTIVE: To evaluate the relationship between levels of soluble Fas (sFas) and soluble Fas ligand (sFasL) in serum and peritoneal fluid of endometriosis-associated infertility. METHODS: The soluble Fas ligand and soluble Fas levels in serum and peritoneal fluid of 20 infertile patients with endometriosis were assessed with enzyme-linked immunosorbent assay, and were compared with 14 infertile patients due to chronic pelvic infectious disease and 16 fertile controls. RESULTS: The sFasL levels were significantly higher in infertile patients with endometriosis (175.09 +/- 80.55 pg/mL in serum and 284.50 +/- 152.38 pg/mL in peritoneal fluid) than those of infertile controls (88.47 +/- 43.55 pg/mL in serum and 17.30 +/- 9.62 pg/mL in peritoneal fluid) and fertile controls (16.13 +/- 11.75 pg/mL in serum and 8.84 +/- 2.31 pg/mL in peritoneal fluid). In contrast, as for the sFas levels, infertile patients with endometriosis (828.60 +/- 429.65 pg/mL in serum and 349.61 +/- 288.89 pg/mL in peritoneal fluid) did not show any significant difference compared with those in infertile patients resulting from pelvic infectious disease (868.75 +/- 570.48 pg/mL in serum and 181.76 +/- 157.78 pg/mL in peritoneal fluid) and fertile control (822.26 +/- 129.12 pg/mL in serum and 318.42 +/- 145.16 pg/mL in peritoneal fluid). CONCLUSIONS: Based upon these results, high level of sFasL in serum and peritoneal fluid and thus apoptosis mediated by it may be implicated in the mechanism involved in endometriosis-related infertility.

Association of CA repeat polymorphism in estrogen receptor beta gene with postmenopausal osteoporosis in Chinese.

Postmenopausal osteoporosis (PMO) is considered a polygenic disease. The estrogen receptor beta (ESR2) gene is a candidate mediating the genetic influence on bone mass and the risk of osteoporosis. The aim of this study is to investigate the association of a cytosine-adenine (CA) repeat polymorphism in the fifth intron of the ESR2 gene with PMO in Chinese Han population. The CA repeat polymorphism was genotyped in a case-control study, involving 78 femoral neck PMO patients vs. 122 controls and 108 lumbar spine (L2-4) PMO patients vs. 92 controls. The (CA)(n) < 22 and (CA)(n) > or = 22 alleles were designated short (S) and long (L), respectively. ESR2 genotype was categorically defined as SS (2 S alleles), SL (having the mixed S and L alleles), and LL (2 L alleles). At both the femoral neck and the L2-4 region, LL genotype and L allele frequencies of the PMO group were significantly higher than those of the control group (P < 0.01). The subjects with the SL, the LL, and the combined SL and LL genotype had a significant increased risk of PMO when compared with those with the SS genotype (P < 0.05). After adjustments for age, years since menopause, menopausal age, and body mass index, logistic regression analysis showed that the subjects with the combined SL and LL genotype had increased risk of PMO when compared with those with the SS genotype both at the femoral neck (adjusted OR 4.923, 95%CI 1.986-12.203, P=0.001) and the L2-4 (adjusted OR 2.267, 95%CI 1.121-4.598, P=0.023). This extensive association study has identified the ESR2 CA repeat polymorphism to be independently associated with PMO at the femoral neck and the L2-4 in Chinese Han population. The data also suggested that the presence of the L allele may dominantly increase the risk of PMO at the two regions.