[Prenatal diagnosis and genetic analysis of a fetus with Miller-Dieker syndrome].

OBJECTIVE: To explore the genetic basis for a fetus with lissencephaly. METHODS: Genomic DNA was extracted from amniotic fluid sample and subjected to copy number variation (CNV) analysis. RESULTS: The fetus was found to harbor a heterozygous 5.2 Mb deletion at 17p13.3p13.2, which encompassed the whole critical region of Miller-Dieker syndrome (MDS) (chr17: 1-2 588 909). CONCLUSION: The fetus was diagnosed with MDS. Deletion of the PAFAH1B1 gene may account for the lissencephaly found in the fetus.

[Analysis of ASXL3 gene variant in a child with Bainbridge-Ropers syndrome].

OBJECTIVE: To explore the genetic basis for a child affected with Bainbridge-Ropers syndrome. METHODS: Genomic DNA was extracted from peripheral venous blood samples from the patient and his parents. Whole exome sequencing (WES) was carried out to detect genetic variant of the proband. Candidate variant was verified by Sanger sequencing. RESULTS: The 3-year-old boy presented with psychomotor retardation, linguistic difficulties, mental retardation and peculiar craniofacial phenotype. A de novo heterozygous nonsense variant of the ASXL3 gene, c.3106C>T, was identified by WES in the proband, and the same mutation was not found among his parents. Based on the American College of Medical Genetics and Genomics standards and guidelines, the c.3106C>T variant was predicted to be pathogenic (PVS1+PS2+PP4). CONCLUSION: The heterozygous variant c.3106C>T of the ASXL3 gene probably underlies the Bainbridge-Ropers syndrome in the patient. Above result has enabled the clinical diagnosis and genetic counseling for the family.

[Clinical and genetic analysis of a child with X-linked hypohidrotic ectodermal dysplasia].

OBJECTIVE: To explore the clinical and genetic characteristics of a child with X-linked hypohidrotic ectodermal dysplasia (XLHED). METHODS: Clinical data of the child was collected. Peripheral blood samples were taken from the child and his parents with informed consent and subjected to copy number variation (CNV) analysis and whole exome sequencing (WES). RESULTS: The male infant manifested sparse hair, anhidrosis, anuresis due to polycystic kidney dysplasia, external genital malformation and anal atresia. WES has revealed a 406 bp hemizygous deletion at Xq13 (68 836 147-68 836 553) in the proband, which encompassed exon 1 of the EDA gene. A heterozygous deletion at the same site was detected in the mother, while no deletion or duplication of the site was detected in the father. CONCLUSION: The hemizygous deletion of EDA gene exon 1 probably underlay the ectodermal dysplasia in the proband. Above result has provided a basis for genetic counseling and prenatal diagnosis for the family.

[Prenatal diagnosis of a fetus with X-linked hypohidrotic ectodermal dysplasia].

OBJECTIVE: To detect variant of EDA gene in a fetus with absence of germ teeth detected by prenatal ultrasonography. METHODS: Clinical data and amniotic fluid and peripheral venous blood samples of the pregnant woman were collected for the analysis. Following extraction of genome DNA, the coding regions of the EDA gene were amplified by PCR and subjected to next-generation sequencing. Candidate variant was verified by Sanger sequencing. RESULTS: The pregnant woman was found to carry a heterozygous c.574G>A variant in the EDA gene, for which the fetus was hemizygous. Bioinformatic analysis suggested the variant to be pathogenic. CONCLUSION: Combined ultrasonographic and genetic findings suggested the fetus is affected with X-linked hypohidrotic ectodermal dysplasia due to pathogenic variant of the EDA gene.

[Genetic diagnosis for a patient with Leydig cell hypoplasia caused by two novel variants of LHCGR gene].

OBJECTIVE: To explore the genetic basis for a patient with Leydig cell hypoplasia. METHODS: Whole exome sequencing was used to detect genetic variants in the patient. Suspect variants were verified by PCR and Sanger sequencing of the family members. RESULTS: The patient was found to carry two novel variants, namely c.265A>T (p.Ile189Leu) and c.422T>C (p.Val141Ala), of the luteinizing hormone receptor gene (LHCGR), where were respectively inherited from her father and mother. Upon prenatal diagnosis, the fetus was found to be a heterozygous carrier of the c.265A>T (p.Ile189Leu) variant. CONCLUSION: The compound heterozygous variants of c.265A>T (p.Ile189Leu) and c.422T>C (p.Val141Ala) of the LHCGR gene probably underlie the Leydig cell hypoplasia in the patient.

Follicular fluid Aß40 concentrations may be associated with ongoing pregnancy following in vitro fertilization.

PURPOSE: To determine whether Aß40 levels in the follicular fluid (FF) of infertile women undergoing IVF demonstrate a relationship with IVF cycle parameters and outcome. METHODS: FF Aß40 levels were compared between patients achieving ongoing pregnancy and those with unsuccessful cycles. Clinical data such as ongoing pregnancy rate, implantation rate, number of oocytes retrieved, number of 8 cells embryos with ≤5 % fragmants, ratio of 8 cells embryos with ≤5 % fragmants to total embryos per patient and cleavage rate were compared among three percentile groups of Aß40. CCK-8 method was used to measure the effect of Aß40 on rat granulosa cells proliferation in vitro. RT-PCR was used to detect the mRNA expression levels of steroidogenesis related genes. RESULTS: Patients achieving ongoing pregnancy (n = 26; 50.98%) demonstrated significantly higher FF Aß40 levels compared to those with unsuccessful cycles (n = 25; 49.02%; P = 0.024). No significant differences were observed in APP (amyloid precursor protein) and its other proteolysis products including sAPPα, sAPPßand Aß 42 between the two groups. Statistically significant differences between the three percentile groups of Aß 40 were observed only in the implantation rates and ongoing pregnancy rates. There were no statistically significant differences between the three percentile groups in the age, No. oocytes retrieved, No. 2 pronucleus, No. embryos transferred, No. 8 cells embryos with ≤5% fragmants and cleavage rate. Significantly negative correlation exists between APP and AFC (antral follicle count) (R =-0.360, P = 0.005) and oocytes retrieved (R =-0.378, P = 0.004). There were also significantly positive correlations between Aß40 and Aß42 (R = 0.407, P = 0.000), between AFC and oocytes retrieved (R = 0.476, P = 0.000). Rat granulosa cells treated with Aß40 of different concentrations have improved their proliferative ability. Cells treated with 200 pg/ml Aß40 have the strongest ability of proliferation. 200 pg/ml Aß40 enhanced the expression of key molecules during steroidogenesis such as IGF-1,IGF-1receptor (IGF-1R),FSH receptor (FSHR),P450 aromatase (P450arom),steroidogenic acute regulatory protein (StAR) and cholesterol side-chain cleavage cytochromes P450(P450scc). CONCLUSIONS: Aß40 levels in follicle fluid may be associated with ongoing pregnancy and the moderate expression level of Aß40 is important for oocytes and embryos development.

[Effect of retinoic acid on alkaline phosphatase expression in rat adipose-derived stem cells].

OBJECTIVE: To investigate whether adipose-derived stem cells (ADSCs) induced by retinoic acid (RA) in vitro express primordial germ cell marker alkaline phosphatase (ALP) and vasa. METHODS: ADSCs were isolated from adult female SD rats and cultured in vitro. The third passage of ADSCs was identified by adipogenic differentiation, osteogenic differentiation and cell surface marker detection. The ADSCs were treated with 1×10(-5), 1×10(-6), or 1×10(-7) mol/L RA for 7 or 14 days, and the cellular expression of ALP was detected. vasa mRNA expression in ADSCs treated with 1×10(-5) mol/L RA for 7 days was detected using RT-PCR. RESULTS: The OD value of ADSCs treated with 1×10(-5), 1×10(-6), or 1×10(-7) mol/L RA was 0.59∓0.04, 0.27∓0.07, and 0.15∓0.03 after a 7-day treatment, and was 0.42∓0.02, 0.34∓0.01, and 0.19∓0.02 after a 14-day treatment, respectively, demonstrating significantly enhanced ALP expression in RA-treated ADSCs compared with that in the control cells (0.07∓0.01 and 0.07∓0.01 at 7 and 14 days, respectively, P<0.01). The ADSCs showed a negative vasa mRNA expression after 1×10(-5) mol/L RA treatment for 7 days. CONCLUSION: RA-induced ADSCs express ALP, a marker of primordial germ cells, but does not express the primordial germ cell marker vasa.

Paravertebral fascial massage promotes brain development of neonatal rats via the insulin-like growth factor 1 pathway.

Massage in traditional Chinese medicine can promote body and brain development of premature and normal newborn infants. In the present study, neonatal rats (1 day old) underwent paravertebral fascial massage (15 consecutive days), followed by subcutaneous injection of insulin-like growth factor 1 receptor antagonist, JB1 (9 consecutive days). Paravertebral fascial massage significantly increased insulin-like growth factor 1 expression and cell proliferation in the subventricular zone of the lateral ventricle and dentate gyrus of the hippocampus. However, JB1 inhibited this increase. Results suggest that paravertebral fascial massage can promote brain development of neonatal rats via the insulin-like growth factor 1 pathway.

[Effect of allogenic adipose-derived stem cell grafting on serum biochemical indicators and sporting ability of endurance trained rats].

OBJECTIVE: To observe the effect of transplantation of allogeneic adipose-derived stem cells (ADSCs) on serum biochemical indicators and sporting ability in highly endurance-trained rats from a fasciological perspective. METHODS: The ADSCs were cultured in vitro. Forty male Wistar rats were randomized into 4 equal groups, namely the blank control group, overtraining (model) group, transplantation without training group and overtraining plus transplantation group. The rats in the two overtraining groups were subjected to exhaustive swimming for 1 week, and in the two transplantation groups, cultured allogeneic ADSCs (2×10(6)/ml) were injected via the tail vein. The exhaustion time in swimming and the serum levels of BUN, LDH, BLa, and Hb of the rats were recorded after the treatments. RESULTS: The rats in the model group showed significantly increased serum BUN, LDH and BLa levels and decreased Hb level with a extended exhaustion time as compared with those in the blank control group (P<0.01). The BUN, LDH and BLa was significantly lower, Hb level higher and the exhaustion time significantly longer in the overtraining plus transplantation group than those in the model group (P<0.01). CONCLUSIONS: ADSCs can effectively prolong the exhaustion time of rats during exhaustive swimming and enhance their sporting ability.