Shi Yc

Bone marrow mesenchymal stem cells reduce intestinal ischemia/reperfusion injuries in rats.

BACKGROUND Adult stem cells are promising novel therapies in regenerative medicine. We investigated effects of bone marrow-derived mesenchymal stem cells (MSCs) on intestinal mucosal permeability impaired by ischemia/reperfusion (I/R). METHODS We used a common I/R model in rats to induce intestinal injury by clamping and unclamping the superior mesenteric artery (SMA) in female Sprague-Dawley rats. MSCs were directly injected into the small intestinal submucosa of the syngenic female rats. Control group were injected with the same volume of 0.9% sodium chloride. Small intestine samples were examined for the engraftment of donor-derived mesenchymal stem cells (MSCs) by Y chromosome in situ hybridization analysis. The small intestinal permeability and histomorphologic alternations were measured to evaluate the therapeutic effect of MSCs transplantation. RESULTS Small intestinal permeability and villi injuries were significantly reduced in the MSCs administrated group compared with control group. MSCs administration accelerated the recovery of the intestinal barrier dysfunction. CONCLUSION We concluded that submucosal infusion of MSCs might exert protective effects on the integrity of intestinal barrier.

A novel splice site mutation in the dentin sialophosphoprotein gene in a Chinese family with dentinogenesis imperfecta type II.

Twenty-four individuals were investigated that spanned six generations in a Chinese family affected with an apparently autosomal dominant form of dentinogenesis imperfecta type II (DGI-II, OMIM #125490). All affected individuals presented with typical, clinical and radiographic features of DGI-II, but without bilateral progressive high-frequency sensorineural hearing loss. To investigate the mutated molecule, a positional candidate approach was used to determine the mutated gene in this family. Genomic DNA was obtained from 24 affected individuals, 18 unaffected relatives of the family and 50 controls. Haplotype analysis was performed using leukocyte DNA for 6 short tandem repeat (STR) markers present in chromosome 4 (D4S1534, GATA62A11, DSPP, DMP1, SPP1 and D4S1563). In the critical region between D4S1534 and DMP1, the dentin sialophosphoprotein (DSPP) gene (OMIM *125485) was considered as the strongest candidate gene. The first four exons and exon/intron boundaries of the gene were analyzed using DNA from 24 affected individuals and 18 unaffected relatives of the same family. DNA sequencing revealed a heterozygous deletion mutation in intron 2 (at positions -3 to -25), which resulted in a frameshift mutation, that changed the acceptor site sequence from CAG to AAG (IVS2-3C-->A) and may also have disrupted the branch point consensus sequence in intron 2. The mutation was found in the 24 affected individuals, but not in the 18 unaffected relatives and 50 controls. The deletion was identified by allele-specific sequencing and denaturing high-performance liquid chromatography (DHPLC) analysis. We conclude that the heterozygous deletion mutation contributed to the pathogenesis of DGI-II.

A rare Y chromosome constitutional rearrangement: a partial AZFb deletion and duplication within chromosome Yp in an infertile man with severe oligoasthenoteratozoospermia

We report a case of an infertile man with severe oligoasthenoteratozoospermia with a partial azoospermia factor b (AZFb) deletion and duplication region within chromosome Yp11.2. The hormonal profile was normal for serum concentrations of follicle-stimulating hormone, luteinizing hormone, testosterone and oestradiol. The patient, who showed a 46,XY karyotype, had an approximate 2.4 Mb inherited duplication region in Yp11.2 and a de novo partial AZFb deletion, which spanned 5.25 Mb including eight protein coding genes and four non-coding transcripts, but did not remove the RBMY gene family. Both proximal and distal breakpoints of the deletion were outside any palindromic region or inverted repeat sequence and intra-chromosomal non-allelic homologous recombination could not have been the deletion mechanism. The partial AZFb deletion in our case diminished sperm production, but did not completely extinguish spermatogenesis. Considering severe oligozoospermia, spermatozoa in the patients ejaculate were used for intracytoplasmic sperm injection, resulting in two twin pregnancies.

Association between ubiquitin-specific protease USP26 polymorphism and male infertility in Chinese men.

BACKGROUND Increased sperm ubiquitin was inversely associated with sperm count and motility. Ubiquitin-specific protease 26 (USP26), which is an X-linked gene, has been studied as a potential infertility gene. There are conflicting reports on whether variations in USP26 are associated with spermatogenesis. METHODS In order to assess that USP26 polymorphisms contribute to male infertility, we screened 221 infertile men with azoospermia, oligozoospermia, asthenozoospermia, or oligoasthenozoospermia, and 101 control fertile men using DNA sequencing. RESULTS There were six polymorphisms identified, including an unreported variation (508G>A, G170R). Only the allele frequency of 576G>A was significantly higher in fertile men than infertile patients (p<0.001), although this variant does not result in an amino acid change. The major haplotypes in fertile and infertile men were TGATC (76.2% vs 47.5% of the population, p<0.001) and TGGTC (14.9% vs 39.4%, p<0.001). The haplotype TGATC was under-transmitted, whereas the haplotype TGGTC was over-transmitted in infertile men with asthenozoospermia and oligoasthenozoospermia. CONCLUSIONS Our results indicated the variation of USP26 was not directly associated with human sperm count but suggested it might be a potential role in sperm motility. The 576G>A synonymous single nucleotide polymorphism (SNP) might have a role in improving the sperm motility.

A girl with distinctive features of borderline high blood pressure, short stature, characteristic brachydactyly, and 11.47 Mb deletion in 12p11.21–12p12.2 by oligonucleotide array CGH

Interstitial deletions of the short arm of chromosome 12 are rare. Since Tenconi et al. [1975] described a male baby with a de novo interstitial deletion of 12p, a total of 10 additional patients have been reported [Orye and Craen, 1975; Magenis et al., 1981; Boilly-Dartigalongue et al., 1985; Fryns et al., 1990; Nagai et al., 1995; Gl€aser et al., 2003; Stumm et al., 2007]. Interstitial deletion of 12p could be divided into four groups according to the region of deletion: (1) deletions including the p1! p11; (2) deletions extending more distally (p13! p11); (3) deletions in band 12p13, and (4) deletions encompassing the terminal of 12p [Gl€aser et al., 2003]. Common phenotypic findings constituting the interstitial deletions of 12p include mental retardation, developmental delay, craniofacial anomalies, microcephaly, brachydactyly, and clinodactyly. Only one patient with borderline high blood pressure, short stature, brachydactyly, and a deletion in 12p11.21! 12.2 region was reported by Nagai et al. [1995]. We encountered a 13-year-old girl with moderate mental retardation, short stature, and characteristic brachydactyly. She was born at term of gestation with normal birth weight of 3,300 g and short stature of 46 cm ( 2.4 SD). The girl spoke at the age of 2 years and walked without assistance at the age of 3 years. Clinical observation showed a small slender girl with height 126 cm ( 4.4 SD), weight 25 kg ( 2.66 SD), borderline high blood pressure (120/80 mmHg), round face, strabimus and chaotic tooth arrangement (Fig. 1). The fifth toe overlapped the fourth one and both of them showed brachydactyly. Serum hormone levels of GH, TSH, T3, and T4 were normal for her age. Cerebral CT scan and MRI were normal. Roentgenograms of hands and feet disclosed the bilateral brachydactyly, with shortened metacarpals of digits 3–5, middle phalanges of digit 5, and metatarsals of digits 4 and 5 (Fig. 2). Cone-shaped epiphyses were not evident. No obvious skeletal abnormalities were found in vertebrae, pelvis, and other tube bones. Now she is enrolled in a special school for children with mental retardation. She can count numbers to 10. Chromosome analysis of G-banding at 400 band showed a possible deletion of 12p11.2 in all metaphases analyzed (Fig. 3A). Family history was negative. Chromosome analysis of both parents showed normal karyotypes, indicating a de novo deletion. This research was approved by Ethics Committee of the hospital, and the written informed consent was obtained from the patient and the parents. Multicolor fluorescent in situ hybridization (M-FISH) analysis using the Spectra Vysion WCP Probe (Vysis, Downers Grove, IL) was performed on metaphases. A reciprocal translocation was excluded. To refine the extent of the deletion on the molecular cytogenetic level, analysis of using a genomic-wide high-density oligo array (OaCGH44K) with 44,000 probes covering more than 30,000 mapped genes was carried out according to Agilent manufacturer’s procedures and statistical algorithms [Fan et al., 2007; www.agilent. com.chem/gocgh]. An 11.47 Mb interstitial deletion at genomic position 20, 724, 852 bp! 32, 201, 544 bp in

Rapid Molecular Prenatal Diagnosis of Spondyloepiphyseal Dysplasia Congenita by PCR-SSP Assay

Heterozygous mutations of COL2A1 gene are responsible for type II collagenopathies. The common skeletal phenotypes include achondrogenesis type II, hypochondrogenesis, Stickler dysplasia, Kniest dysplasia, late onset spondyloepiphyseal dysplasia, and spondyloepiphyseal dysplasia congenita (SEDC). Prevention of SEDC can be achieved by prenatal diagnosis. This study reports the first rapid molecular prenatal diagnosis of SEDC performed in China by polymerase chain reaction sequence-specific primer (PCR-SSP) analysis. The pregnant woman we previously reported with SEDC carried the G to A substitution at nucleotide 1510 in exon 23 of COL2A1 gene, which caused a change from glycine to serine at codon 504 (G504S). By the time the woman got pregnant again, she had terminated two pregnancies and still had no child. In the first pregnancy, the molecular mutation of the family was not yet identified, and therefore prenatal diagnosis was unable to be performed by DNA analysis. In the second pregnancy, G504S mutation was found from fetal DNA. At the time of her third pregnancy, the woman and her husband became extremely worried about the potential SEDC for the fetus. For this reason, a quick and reliable molecular prenatal diagnosis of SEDC was performed by a PCR-SSP on an amniocyte sample collected at the 14th week of pregnancy. No mutation of the fetal DNA was identified. The result was obtained within 24 h after the sample was collected. The technique could be applied in confirmatory diagnosis and prenatal diagnosis for the affected family.

A novel insertion mutation in the SEDL gene results in X-linked spondyloepiphyseal dysplasia tarda in a large Chinese pedigree

BACKGROUND Spondyloepiphyseal dysplasia tarda (SEDT) is an X-chromosome linked primary skeletal dysplasia characterized by a disproportionate short-trunked short stature, dysplasia of the large joints and flattened thoracic and lumber vertebral bodies. The objective of this study is to describe a large Chinese SEDT family with a milder phenotype and describe the molecular and clinical findings. METHODS Eight affected males of the family were diagnosed with SEDT according to their clinical and radiological features. Direct DNA sequencing of the SEDL gene was performed. RT-PCR experiments on total RNA from blood lymphocytes were performed to confirm the defect on the SEDL gene. A short summary of all currently known SEDL gene mutations is presented. RESULTS DNA sequencing revealed that all the affected males carried an insertion mutation (c.370-371insA) unreported previously, predicted to result in frameshifts and generate a premature stop codon (p.S124fsX127). The identical mutation was also observed in a 10-year old presymptomatic boy of the family. Eight female carriers had the typical sequencing chromatograms of heterozygotes. CONCLUSIONS Identification of the novel insertion mutation (c.370-371insA) in this SEDT family enables carrier detection and presymptomatic/prenatal diagnosis, but also the detailed molecular and clinical features will be useful for extending the evidence for genetic and phenotypic heterogeneity in SEDT.

Meiotic pairing error in an infertile male bearing reciprocal deletion of chromosome 13.

A series of complex processes takes place during the first meiotic division, including pairing, synapsis, recombination, and segregation of homologous chromosomes. When any of these processes is altered, cellular checkpoints arrest the progression of meiosis and induce cell loss, causing a severe reduction in fertility, or even sterility. In this study, we report on a 29-year-old, healthy, but severe oligozoospermic male with a supernumerary, ring-neocentric 13q12.3 → 13q22 chromosome and a reciprocal deletion, which interfere with the meiotic pairing of chromosomes 13, causing spermatogenesis failure.

Erratum to “A G560S mutation in α1 (I) collagen causes familial osteogenesis imperfecta type IV” [Clinica Chimica Acta 409 (2009) 145–146]

The Publisher regrets that the following error has occurred in the above article. Subsequently, a heterozygous substitution of 7872 g. G>A (AF017178.2) in exon 25 of COL1A1 was found by DNA sequencing in all affected individuals and was further confirmed by DHPLC analysis. However, to our knowledge, the mutation reported here had been proposed only by Mackay et al. [5] who reported a sporadic case in a six-year-old child without detailed clinical description.