[Phenotypic and genetic analysis of an inv dup(15) case with a BP3:BP3 rearrangement].

Objective: To analyze a case of supernumerary marker chromosome (SMC) with combined genetic techniques and explore its correlation with the clinical phenotype.

Methods: The SMC was analyzed with G-banded karyotyping, multiplex ligation dependent probe amplification (MLPA), fluorescence in situ hybridization (FISH), and single nucleotide polymorphism array (SNP-array).

Results: G-banding analysis indicated that the patient has a karyotype of 47,XX,+mar.

[Analysis of AR gene mutation in a family affected with complete androgen insensitivity syndrome using long chain RT-PCR].

Objective: To identify potential mutation of androgen receptor (AR) gene in a patient with complete androgen insensitivity syndrome (CAIS) and his family members.

Methods: Total RNA and genomic DNA were extracted from the peripheral blood samples derived from the proband and her family members. Sequences of 7 exons of the AR gene were amplified with reverse transcriptase PCR(RT-PCR) and subjected to direct sequencing.

Whole-exome sequencing identifies compound heterozygous mutations in ARSA of two siblings presented with atypical onset of metachromatic leukodystrophy from a Chinese pedigree.

Background: Metachromatic leukodystrophy (MLD) is a rare inherited lysosomal storage disorder caused mainly by variants in arylsulfatase A (ARSA) gene. MLD can be divided into three major clinical forms according to the age of onset: late infantile, juvenile, and adult. We report two siblings of late infantile MLD presenting with cerebellar ataxia as the only first clinical symptom.

[Cytogenetic and molecular genetic analysis of a case with mosaic marker chromosomes].

Objective: To explore the source of small supernumerary marker chromosome in a case.

Methods: G-banded karyotyping, fluorescence in situ hybridization, multiple sequence tagged sites (STS) of the Y chromosome, and Illumima Human Cyto SNP-12 Beadchip analysis were carried out.

Results: The karyotype was mos 46,X,+mar1[21]/46,X,+mar2[78].

[Genetic diagnosis and analysis of related genes for a pedigree with 2p25 and 12p13 cryptic rearrangements].

Objective: To analyze chromosome aberration in a child with mental retardation and abnormalities and its parents.

Methods: Chromosome G banding, multiplex ligation-dependent probe amplification, fluorescence in situ hybridization and single nucleotide polymorphisms array were employed for analysis.

Results: Karyotype analysis revealed that the child was 46,XX and the father was 46,XY, while the mother was 46,XX, add (12)(p13).

[Analysis of small supernumerary marker chromosome 15q11 in four infertile males].

Objective: To delineate the origins of small supernumerary marker chromosomes (sSMCs) identified in 4 infertile males.

Methods: The sSMCs were analyzed with combined G-banding, N-banding, multiplex ligation-dependent probe amplification (MLPA), fluorescence in situ hybridization (FISH) and single nucleotide polymorphisms array (SNP-array) techniques.

Results: G-banding analysis has suggested a 46,X,-Y,+mar karyotype in all of the 4 cases.

[Delineation of three structural Y chromosome aberrations combined molecular techniques].

Objective: To delineate the structure of Y chromosome aberrations and recombinant mechanisms for three patients.

Methods: Karyotype analysis, multiplex ligation dependent probe amplification (MLPA), fluorescence in situ hybridization (FISH), Y chromosome sequence tagged sites (STS) analysis, human whole genome-wide SNP array were used.

Results: The karyotypes of the three patients were 46, X, +mar.

Sleep duration and its correlates in middle-aged and elderly Chinese women: the Shanghai Women's Health Study.

Background: Abnormal sleep duration, either long or short, is associated with disease risk and mortality. Little information is available on sleep duration and its correlates among Chinese women.

Methods: Using information collected from 68,832 women who participated in the Shanghai Women's Health Study (SWHS), we evaluated sleep duration and its correlations with sociodemographic and lifestyle factors, health status, and anthropometric measurements and their indexes using polynomial logistic regression.

[Partial AZfc region deletions of the Y chromosome in spermatogenic dysfunction patients].

Objective: To investigate the influence of partial deletions in the AZFc region of the Y chromosome on spermatogenesis.

Methods: We selected 9 sequence tagged sites (sY1258, sY1291, sY254, sY255, sY1201, sY1206, sY1161, sY1197 and sY1191) in the AZFc region of the Y chromosome, with ZFX/ZFY and SRY (sY14) as the interior control. We amplified by multiplex PCR the DNA of 160 patients with azoospermia or severe oligozoospermia that showed no microdeletion of the Y chromosome (the case group) and another 76 males with normal fertility (the control group).

A rapid and sensitive protocol for prenatal molecular diagnosis of X-linked adrenoleukodystrophy.

Background: X-linked adrenoleukodystrophy (X-ALD) is a neurodegenerative genetic disease characterized by progressive demylination of the brain, adrenal insufficiency and elevated VLCFA level. ABCD1gene is the disease gene and more than 500 unique mutations in the ABCD1gene have been recorded in the database, approximately 60% of which are noncurrent ones. Although great progress has been made in the treatment of X-ALD, prenatal diagnosis is still badly needed by X-ALD-stricken families.

[Breakpoint localization of Y-chromosome massive deletions in 49 spermatogenesis dysfunction patients].

Objective: To analyze the characteristics of azoospermia factor(AZF) deletions in Y-chromosome.

Methods: Based on the AZF microdeletion screening on 272 cases of azoospermia and 240 cases of severe oligozoo spermia, 49 cases were investigated using 23 sequence-tagged sites (STS) in AZFa, AZFb and AZFc. For some cases, single nucleotide rarians (SNV) method was applied to identify the single nucleotide polymorphism (SNPs) in four DAZ gene copies and to determine the copy number of the DAZ gene.

[Molecular diagnosis of spinal muscular atrophy by multiplex ligation-dependent probe amplification].

Objective: To investigate the effect of multiplex ligation-dependent probe amplification (MLPA) in molecular diagnosis of spinal muscular atrophy (SMA).

Methods: Peripheral blood samples were collected from 13 SMA patients, 31 parents of SMA patients, 50 healthy individuals without family history of SMA, and 10 specimens of amniotic fluid from these families were collected too. Genomic DNA was analyzed by MLPA, conventional PCR-RFLP, and allele-specific PCR.

Evaluation of an in-house protocol for prenatal molecular diagnosis of SMA in Chinese.

Background: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder characterized by degeneration of the anterior horn of the spinal cord, leading to symmetric muscle weakness and atrophy. About 95% of SMA patients have homozygous loss of SMN1 which can be detected by conventional PCR-RFLP testing. However, the method cannot distinguish heterozygous healthy carriers.

Construction of sperm-specific lactate dehydrogenase DNA vaccine and experimental study of its immunocontraceptive effect on mice.

Lactate dehydrogenase C4 (LDHC4) is a key enzyme for sperm metabolism. It is distributed specifically in testis and is highly immunogenic. In this study, two DNA vaccines pVAX1-hLDHC and pVAX1-mLDHC were constructed by inserting coding sequences of human and mice LDHC4 into the eukaryotic expression vector pVAX1.

[Prenatal molecular diagnosis of four fetuses at high risk for X-linked adrenoleukodystrophy].

Objective: To investigate methods for prenatal molecular diagnosis of fetuses at high risk for X-linked adrenoleukodystrophy (X-ALD).

Methods: The amniotic fluid was obtained and genomic DNA was isolated from amniotic fluid cells. Maternal contamination was evaluated by paternity test.

[Prokaryotic expression and characterization of N-terminal truncated glycoprotein gp85 of Epstein-Barr virus].

Aim: To construct a prokaryotic recombinant vector of Epstein-Barr virus (EBV) membrane protein gp85, to express the protein in E.coli and characterize the antigenicity of this non-glycosylated protein.

Methods: The BXLF2 gene coding 5'-terminal truncated of EBV gp85 was amplified from the EBV strain B95-8 cell line with specific primers.

Characterization of a Cys329Gly mutation causing hereditary factor VII deficiency.

We have previously reported a homozygous Cys329Gly mutation in a Chinese patient with factor VII (FVII) deficiency. Others have found a heterozygous Cys329Gly mutation in the F7 gene from patients of three different pedigrees. However, none of the reports included the expression and characterization of the mutant FVII in vitro.

Prenatal molecular diagnosis of adrenoleukodystrophy.

Objective: To carry out prenatal diagnosis on two fetuses of different pedigrees with X-linked adrenoleukodystrophy (ALD).

Methods: The amniotic fluid was obtained with the help of a clinical doctor and the genomic DNA was isolated from it. Maternal DNA contamination was excluded by fluorescent STR profiling, The R617G mutation found in the first pedigree was searched in genomic DNA of amniotic fluid cells (AFC) from fetus 1 by amplification refractory mutation system (ARMS) and dot DNA hybridization while the P534R mutation found in pedigree 2 was analyzed in the AFC genomic DNA of fetus 2 by restrictive digestion with Hae II and DNA direct sequencing.

[Expression and characterization of subunit C of mouse lactate dehydrogenase in Escherichia coli].

Objectives: To construct a prokaryotic recombinant vector for mouse lactate dehydrogenase-C and to detect its expression in BL21.

Methods: The coding sequence of mouse lactate dehydrogenase subunit C was amplified from mouse testis RNA with specific primers, and cloned into pGEX-2T after the restriction digestion with BamH I and EcoR I. GST fusion protein was expressed after induction with IPTG.