A novel splicing mutation in intron 2 of DSPP gene in a family with dentinogenesis imperfecta type Ⅱ / 临床检验杂志

Objective To report a familial dentinogenesis imperfecta type Ⅱ (DGI type Ⅱ) with a novel splicing mutation in DSPP (dentin sialophosphoprotein) gene.Methods Based on the result of linkage analysis performed previously to map the candidate gene DSPP in the family, the promoter,the first four exons and exon-intron boundaries of DSPP were directly sequenced for the members of the DGI type Ⅱ family. Denaturing high performance liquid chromatography (DHPLC) analysis was performed to confirm the results of sequencing.Results A novel splicing mutation of 23 bp deletion in intron 2 of DSPP gene was identified by DNA sequence analysis. The mutation changed acceptor site sequence from CAG to AAG, and might result in functional abolition of possible branch point site in intron 2. DHPLC result was consistent with that of sequencing. The mutation may be identified in all affected individuals, but not found in normal members of the family and 50 controls.Conclusion These results suggest the deleted mutation of DSPP gene causes DGI type Ⅱ in the family. The mutation has not been reported before.

Segregation of sex chromosomes in the spermatozoa of 46, XY/47, XXY patients with oligozoospermia by dual fluorescence in-situ hybridization / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the segregation of sex chromosomes in the spermatozoa of a 46, XY/47, XXY patient with oligozoospermia.</p><p><b>METHODS</b>The number of X and Y chromosomes of the ejaculated spermatozoa from the patient with mosaic 46, XY/47, XXY was analysed by X/Y dual fluorescence in-situ hybridization (FISH).</p><p><b>RESULTS</b>Of the 100 spermatozoa analysed, 97 showed either one X chromosome-specific green signal or one Y-chromosome-specific red Y signal in each spermatozoon and only 3 showed no signal. The frequencies of X- and Y-bearing spermatozoa were 49% and 48% respectively. The ratio of X- to Y-bearing spermatozoa was about 1:1 as expected. There was no statistical difference between the chromosome XX and XY frequencies in each spermatozoon from the patient in comparison with those estimated in the control.</p><p><b>CONCLUSION</b>The spermatozoa of 46, XX/47, XXY mosaic patients have a normal gonosomal complement, which allows infertility treatment to be carried out by ICSI.</p>

The mechanism of tetragametic chimerism in a true hermaphroditism with 46, XX/46 ,XY / 中华男科学杂志

<p><b>OBJECTIVE</b>To report a true hermaphroditism due to a teragametic chimerism and to discuss the pathogenesis of tetragametic chimerism.</p><p><b>METHODS</b>Chromosomal analysis and fluorescence in situ hybridization(FISH) were carried out on the lymphocytes from the blood and on the fibroblasts from the cultured skin and on fibroblasts from two different kinds of gonadal tissues of the patient with ambiguous genitalia respectively. Blood groups, human leukocyte antigen (HLA) haplotyping and 77 short tandem repeat (STR) microsatellite markers were tested. The two kinds of tissues in the gonad were detected by histopathological examination. Blood groups, HLA haplotying and 77 STR microsatellite markers parents of the patient's were also analyzed.</p><p><b>RESULTS</b>Either 46,XX or 46,XY karyotype was found in the lymphocytes of the blood and in the fibroblasts of the cultured skin and of the two different kinds of gonadal tissues. Two X chromosome-specific signals or one X and one Y signal were detected in each interphase nucleus by FISH from the lymphocytes of the blood and the fibroblasts of three different tissue cultures. The karyotype of the 46,XY cell line predominated in all cultures except the cultured-fibroblasts from yellow gonadal tissues. STR marker analysis, ABO grouping and HLA study from the patient were identified a single haplotype in the patient from the mother and two different haplotypes from the father. Two kinds of tissues in the gonad were observed by histopathological examination. The yellow tissue was ovary and the white one was testis.</p><p><b>CONCLUSIONS</b>Histopathological examination and chromosomal analysis combined with FISH are very useful methods for the diagnosis of true hermaphroditism. Blood typing, HLA and short tandem repeat microsatellite markers afford strong evidence for confirming tetragametic chimerism. The mechanism of tetragametic chimerism in true hermaphroditism can be explained by a parthenogenetic division of a haploid nucleu into two identical gametes, followed by fertilization with both X and Y spermatozoa and then developed into an organism.</p>

Advances in gonadal differentiation regulated by SRY / 中华男科学杂志

Gondadal differentiation is genetically determined in humans. Sex is determined when the bipotential embryologic tissues differentiate into testes or ovary. SRY, a gene located on the Y chromosome, triggers a complex genetic cascade leading to testicular differentiation. However, only a minority of 46, XY sex reversal patients can be explained by SRY mutations, suggesting that other genes influencing sex determination are to be discovered. Recent studies show that testis differentiation requires insulin receptor family function in mice. SRY normally requires two distinct NLS-dependent nuclear import pathways to reach sufficient levels in the nucleus for gonadal differentiation.

Detection and potential function of anti-FSH antibodies in patients with spermatogenesis dysfunction / 中华男科学杂志

<p><b>OBJECTIVE</b>To detect the anti-FSH antibody using ELISA, and further probe into the role of anti-FSH in infertile patients.</p><p><b>METHODS</b>The anti-FSH antibody was detected using ELISA in the serum of patients with spermatogenesis dysfunction, of infertile patients with normal sperm density and motility, and of normal fertile males.</p><p><b>RESULTS</b>The positive rate of anti-FSH antibody in the patients with oligospermia and/or asthenospermia [22.4% (22/98)] was significantly higher than that in the normal fertile [4% (2/50)] (P < 0.05) and that in the infertile patients with normal sperm density and motility [6.7% (2/30)] (P < 0.05). The positive rate of anti-FSH antibody in the patients with oligospermia and/or asthenospermia was lower than that in the patients with azoospermia [54.5% (12/22)] (P < 0.05). There was no significant difference in the positive rate between the normal control and the sterile males with normal sperm density and motility.</p><p><b>CONCLUSION</b>The anti-FSH antibody may be an important factor to cause spermatogenesis dysfunction by combining FSH to form immune compound and depress the activation of FSH.</p>

COL2A1 gene mutation of a family with spondyloepiphyseal dysplasia conginita / 医学研究生学报

Objective:To investigate a large Chinese family in which 9 patients over 4 generations were diagnosed with a form of autosomal dominant spondyloepimphyseal dysplasia(SEMD).Mothods:X-Ray radiograph of proand at 18-month showed absence of secondary ossification centra of femoral heads.His father at 24-year presented severe spondyloepiphyseal changes that principally involved the vertebral bodies,the femoral necks and femoral heads and characterized by generalized platyspondyly with thoracolumbar scoliosis,irregular femoral necks,absent ossification of femoral heads,flat acetabular roofs and coxa vara.The other patients had similar clinical and radiological features.Haplotyping was performed with leukocyte DNA for 5 micosatellite repeat markers from chromosome 12 and the result showed COL2A1 gene as a candidate gene.A total of 54 exons and promoter of COL2A1 gene were amplified and sequenced from all patients and available normal relatives.In addition,exon 23 of COL2A1 gene was amplified and sequenced from 10 controls simultaneously.Results:All patients were identified a 1510(G→A) transition in exon 23 of COL2A1 gene that caused a change from a COL2A1 coding region in available glycine to serine at amino acid position 504.No mutation was found in the normal relatives and 10 controls. Conclusion:The mutation of COL2A1 gene is responsible for this form of SEDC of the family.This is the first familial report of SEDC relating to 1510G→A mutation of COL2A1 gene.The detailed clinical radiogram data will be useful for extending the phenotypic spectrum of type Ⅱcollagenopathies.

Progress in studies on molecular genetics of spondyloepiphyseal dysplasia / 医学研究生学报

Spondyloepiphyseal dysplasia(SED) includes a group of disorders that cause deformation of vertebrae and epiphyses following gene mutations.Its main clinical manifestations are short stature(with a disproportionately short-trunk),chest malformation and early-onset joint degeneration.These disorders are broadly categorized into two subtypes: congenita(SEDC) and tarda(SEDT).In the 2006 revision of the International Nosology and Classification of Genetic Skeletal Disorders,372 different conditions were listed,of which 215 were associated with one or more of 140 different genes.SEDC has consistently been shown to correlate with defects in the gene COL2A1 on the long arm of chromosome 12,whose product is needed to form normal type-Ⅱ collagen.The gene responsible for SEDT is SEDL,mapped to the short arm of the X chromosome(Xp22).This paper briefly reviews the progress in the studies of molecular genetics of SEDC,SEDT and other rare forms of SED,which might provide some practical help for genetic and prenatal diagnoses of SED.

The investigation of ethic problem in assisted reproductive techniques / 医学研究生学报

The development and wide application of assisted reproductive techniques(ART) help many couples suffered from infertility and/or sterility to get baby.Meanwhile,the usage of ART led to much ethic problem.We,hence,investigate the strategies to deal with the ethic problem in ART correctly.

A novel RNA-splicing mutation in COL1A1 gene causing osteogenesis imperfecta typeⅠin a Chinese family / 医学研究生学报

A) in COL1A1 gene resulting in OI in a Chinese family. The detailed molecular and clinical features will be useful for extending the evidence for genetic and phenotypic heterogeneity in OI and exploring the phenotype-genotype correlations in OI.

An infertile 45,X male carrying an unbalanced(Y,13) translocation:A clinical cytogenetic and molecular study / 医学研究生学报

Objective: To report a case of azoospermia with a karyotype of 45,X,der(Y)t(Y;13)(q11.2;q12),-13,accompanied with slight bilateral gynecomastia and multiple nodules.Methods: The karyotype was identified by karyotyping and FISH,and the breakpoints of the Y chromosome and the copy number of the BRCA2 gene in 13q12 determined by PCR-STS and DNA polymorphic analysis.The testis and nodule tissues of the patient were obtained for biopsy.Results: FISH confirmed SRY and centromere of the Y chromosome on the questionable 13 chromosome and the karyotype to be 45,X,der(Y)t(Y;13)(q11.1;q12),-13.ish der(Y)(SRY+,DYZ3+,wcp13+).PCR-STS showed the deletion of regions AZFa,b and C,with a breakpoint located inYq11.1 below sY82.No deletion of the BRCA2 gene was observed.The patient was diagnosed with Sertoli cell-only syndrome by testicular biopsy and with angiolipomata by pathological examination of the nodule tissue.Conclusion: The patient's phenotype of complete masculinization could be attributed to presence of the SRY gene,and his azoospermia with small testis to the absence of a fragment from Yq11.1 to Yqter.However,the molecular mechanism of angiolipoma remains unknown.

Effect of removal of submandibular gland on the expression of annexin 5 in rat testis / 医学研究生学报

Objective:To investigate the expression of annexin 5 in rat testis after removing submandibular gland in rats. Methods:On day 14,28 and 42 after the operation,the changes of annexin 5 expression in rat testis were analyzed by Western blot and Immunohistochemistry.Results:Western blot showed that there was a 27.5% and a 35.2% significant increase(P

Prenatal molecular diagnosis of two pregnancies in familial G504S mutation of COL2A1 gene resulting spondylepiphyseal dysplasia congenita / 临床检验杂志

Objective To report the prenatal molecular diagnosis for two gravida in a family with spondylepiphyseal dysplasia congenita(SEDC)caused by G504S mutation of COL2A1 gene.Methods DNA of the two fetuses was extracted from amniotic fluid at the 19+3 and 18+6 weeks of gestation respectively.Direct sequencing of two samples were performed after amplifying exon 23 of COL2A1 containing the potential mutation.The femur length and biparietal diameter of the first fetus were measured by sonographic scans every two weeks from 17+3 weeks to 27+3 weeks of gestation,and for the second fetus these parameters were measured from 16+1 to 19+1 weeks of gestation.Results Sequncing analysis revealed the first fetus and his mother presented the same mutation which is specifically associated with SEDC,but the second fetus did not show the mutation of COL2A1 gene.Biparietal diameters of the both fetuses were appropriate for gestational age.Femur length of the second fetus was normal for gestational age but that of the first fetus was shortened evidently after the 23 week of gestation.The parents of the first fetus determined to terminate the pregnancy.A medical termination was carried out at 27+5 weeks of gestation and a male fetus with a relatively large head and short limbs was delivered.The radiological findings of the fetus were consistent with SEDC including generalized platy spondesand shortened long bones.Conclusions Prenatal molecular diagnosis is important for the fetus with risk of SEDC and useful for genetic counseling.Genotype of fetus with risk of SEDC can be identified before sonographic scan.Molecular genetic analysis in conjunction with sonographic monitoring was helpful in prenatal diagnosis of SEDC.

MITOCHONDRIAL DNA MUTATION OF SPERMATOZOA OF INFERTILE MALE AND CHANGE OF MITOCHONDRIA-ULTRASTRUCTURE / 解剖学报

Objective To study the correlation of mitochondrial DNA mutation of spermatozoa and change of mitochondria-ultrastructure with male infertility. Methods The techniques of PCR and DNA sequence analysis were used to detect MTCYB and MTATP-6 fragments of 76 samples of semen with poor motility from infertile male.Of these samples five were identified with mitochondrial DNA deletion and transmission electron microscopic observations were made. Results Under the electron microscopic observations the 5 samples were all seen with abnormal volume of mitochondria in most spermatozoa tails either small or big, disorderly located and asymmetrically distributed;the axonemal structures of sperm wrapped in layers of mitochondria. These samples of sperm were noticeably different in form from those from fertile male. Conclusion Sperm mitochondria-ultrastructure change were observed in samples of sperm mitochondrial DNA mutation.Sperm mitochondrial mutation and its mitochondria-ultrastructure affect the energy supply of sperms during the process of fertilization which may result in male infertility.;