Genetic mutation analysis of 22 patients with congenital absence of vas deferens: a single-center study†.

Congenital absence of the vas deferens (CAVD), a congenital malformation of the male reproductive system, causes obstructive azoospermia and male infertility. Currently, the cystic fibrosis transmembrane conductance regulator (CFTR) has been recognized as the main pathogenic gene in CAVD, with some other genes, such as adhesion G-protein-coupled receptor G2 (ADGRG2), solute carrier family 9 isoform 3 (SLC9A3), sodium channel epithelial 1 subunit beta (SCNN1B), and carbonic anhydrase 12 (CA12), being candidate genes in the pathogenesis of CAVD. However, the frequency and spectrum of these mutations, as well as the pathogenic mechanisms of CAVD, have not been fully investigated.

Pathogenesis of acephalic spermatozoa syndrome caused by SUN5 variant.

Acephalic spermatozoa syndrome (ASS) is a rare teratozoospermia that leads to male infertility. Previous work suggested a genetic origin. Variants of Sad1 and UNC84 domain containing 5 (SUN5) are the main genetic cause of ASS; however, its pathogenesis remains unclear.

Further identification of a 140bp sequence from amid intron 9 of human FMR1 gene as a new exon.

Background: The disease gene of fragile X syndrome, FMR1 gene, encodes fragile X mental retardation protein (FMRP). The alternative splicing (AS) of FMR1 can affect the structure and function of FMRP. However, the biological functions of alternatively spliced isoforms remain elusive.

Splicing of exon 9a in FMR1 transcripts results in a truncated FMRP with altered subcellular distribution.

FMRP is an RNA-binding protein, loss of which causes fragile X syndrome (FXS). FMRP has several isoforms resulted from alternative splicing (AS) of fragile X mental retardation 1 (FMR1) gene, but their biological functions are still poorly understood. In the analysis of alternatively spliced FMR1 transcripts in the blood cells from a patient with FXS-like phenotypes (normal CGG repeats and no mutation in coding sequence of FMR1), we identified three novel FMR1 transcripts that include a previously unidentified microexon (46 bp), terming the exon 9a.

Bioinformatics analysis of gene expression profiles of dermatomyositis.

Dermatomyositis (DM) is a type of autoimmune inflammatory myopathy, which primarily affects the skin and muscle. The underlying mechanisms of DM remain poorly understood. The present study aimed to explore gene expression profile alterations, investigate the underlying mechanisms, and identify novel targets for DM.

[Advances in the studies of spermatogonial stem cells in primate mammals].

Spermatogonial stem cells(SSCs) are a type of spermatogonial cells that play an important role in the spermatogenesis of males. SSCs not only possess the properties of stem cells but also differentiate into sperm. They are the unique adult stem cells that transmit genetic information to subsequent generations.

Aggregation of Ribosomal Protein S6 at Nucleolus Is Cell Cycle-Controlled and Its Function in Pre-rRNA Processing Is Phosphorylation Dependent.

Ribosomal protein S6 (rpS6) has long been regarded as one of the primary r-proteins that functions in the early stage of 40S subunit assembly, but its actual role is still obscure. The correct forming of 18S rRNA is a key step in the nuclear synthesis of 40S subunit. In this study, we demonstrate that rpS6 participates in the processing of 30S pre-rRNA to 18S rRNA only when its C-terminal five serines are phosphorylated, however, the process of entering the nucleus and then targeting the nucleolus does not dependent its phosphorylation.

[Identification of a mosaic mutation of NF1 gene in a sporadic case of neurofibromatosis type 1].

Objective: To detect NF1 gene mutation in a patient with neurofibromatosis type 1.

Methods: Five fragments encompassing the entire coding sequence of the NF1 gene were amplified with reverse transcription PCR. PCR products were directly sequenced.

[Progress in molecular diagnosis of fragile X syndrome].

Fragile X mental retardation 1 is the gene underlying fragile X syndrome (FXS). Its product, fragile X mental retardation protein, is closely involved with development of brain and neurons. PCR and Southern blotting have been the major methods for laboratory diagnosis of FXS.

Two novel multiple mutations in chinese patients with adrenoleukodystrophy.

X-linked adrenoleukodystrophy (X-ALD) is a neurodegenerative disorder caused by mutations in the ABCD1 gene. Up to now, more than 1 050 mutations have been reported in the ABCD1 gene, of which only 10 are multiple mutations in one allele of the gene. In this study, we report 2 novel multiple mutations in 2 patients with X-ALD from 2 unrelated Chinese families.

[A novel missense mutation resulting in X-linked adrenoleukodystrophy in female heterozygotes of a Chinese family].

Objective: To identify ABCD1 gene mutation in a Chinese family with three heterozygous female patients.

Methods: Four fragments covering the entire coding sequence of the ABCD1 gene from one of the female patients were amplified by reverse transcription-PCR. The PCR products were directly sequenced.

[Effect of OA kneepad on apoptosis genes Bcl-2 and p53 expression in articular cartilage cells of experimental knee osteoarthritis].

Objective: To study the effects of kneepad on expression of Bcl-2 and p53 mRNA of chondrocyte in white rabbits with knee osteoarthritis, so as to explore and treatment mechanism of OA kneepad on apoptosis of chondrocytes of rabbits with knee osteoarthritis in molecular degree.

Methods: Forty-four Japanese healthy 6-month-old rabbits (equal male and female,the weight ranging from 2 to 2.2 kg) were used to establish knee osteoarthritis models by modified Hulth method.

[Mutation analysis of SMN gene in a patient and his family with spinal muscular atrophy].

Objective: To perform mutation analysis and describe the genotype of the SMN gene in a patient with spinal muscular atrophy (SMA) and his family.

Methods: Deletion analysis of the SMN1 exon 7 by conventional PCR-restriction fragment length polymorphism (RFLP) and allele-specific PCR, and gene dosage of SMN1 and SMN2 by multiplex ligation-dependent probe amplification (MLPA) were performed for the patient and his parents; reverse transcriptase (RT)-PCR and sequencing were performed for the patient. To determine whether the SMN variant was exclusive to transcripts derived from SMN1, the RT-PCR product of the patient was subcloned and multiple clones were sequenced directly; PCR of SMN exon 5 from the genomic DNA of the parents and direct sequencing were performed to confirm the mutation.

[Molecular diagnosis of a Chinese pedigree with osteogenesis imperfecta type I].

Objective: To perform molecular diagnosis for a Chinese pedigree with osteogenesis imperfecta type I.

Methods: Thirty pairs of primers were designed to amplify all the 52 exons, exon boundaries and promoter region of the COL1A1 gene from genomic DNA of peripheral blood cells of the family members. The PCR products were purified and directly sequenced.

[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.

[Genetic polymorphism of sperm-specifically expressed genes and male infertility].

The expression of sperm-specific genes is necessary in spermiogenesis, and the genetic polymorphism of these genes may impair spermatogenesis, leading to male infertility. Mamy investigations have been conducted on the polymorphism of some candidate genes that are specifically expressed in spermiogenesis, the relation of genetic polymorphism with male infertility and its possible mechanism. Further attempts have been made at the explanation of the causes of infertility from the genetic angle.

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.

Increased expression of hLRH-1 in human gastric cancer and its implication in tumorigenesis.

Altered signaling pathways or deregulated transcription factors represent an important category of molecular events leading to aberrant gene regulation in gastric cancer, among which the role of WNT/beta-catenin pathway remains unclear. LRH-1 is a critical transcription factor in controlling cell proliferation via crosstalk with the beta-catenin signaling pathway. In order to gain a knowledge of the expression of hLRH-1v1 and hLRH-1 in gastric cancer, a Q-PCR analysis was carried out.

[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.