OBJECTIVE: To explore the pathogenesis for a SRY-negative male with 46,XX disorder of sex development (DSD). METHODS: Peripheral blood samples of the patient and his family members were subjected to chromosomal karyotyping, routine PCR, real-time fluorescence quantitative PCR, whole exome sequencing and whole genome sequencing. The data was analyzed with NextGENe software. RESULTS: Both the proband and his brother presented a 46,XX karyotype with negative SRY gene, while their father presented normal phenotype and karyotype with positive SRY gene. No pathogenic variant associated with sex development was detected by whole exome sequencing, while a 243 kb duplication was detected by whole genome sequencing in the 5' upstream region of the SOX9 gene in the proband, his brother and father. The same duplication was not found in his sister and mother. CONCLUSION The 243 kb duplication at the 5' upstream of the SOX9 gene may predispose to the 46,XX DSD in this family. It is speculated that there exist an unknown core regulatory element in the upstream of the SOX9, and its duplication may trigger expression of SOX9 and initiate testicular differentiation in the absence of SRY gene.
Disorders of Sex Development/genetics* ; Female ; Humans ; Male ; Mutation/genetics* ; Regulatory Sequences, Nucleic Acid/genetics* ; Sex-Determining Region Y Protein/genetics* ; Testis ; Whole Exome Sequencing

Adult ; Animals ; Female ; Frameshift Mutation/genetics* ; Genes, sry ; Gonadal Dysgenesis, 46,XY/genetics* ; Humans ; Menarche ; Ovary/pathology* ; Oviducts/pathology* ; Sex-Determining Region Y Protein/genetics*

ObjectiveTo identify the etiology of chromosome abnormality in an infertile man and analyze the correlation between the genotype and phenotype.

METHODSWe analyzed the karyotype of an infertile male using the routine G-banding technique and then the chromosome abnormality of the patient by Illumina Human CytoSNP-12 Beadchip array.

RESULTSNegative results were found in the examination of the sex-determining region Y (SRY) gene and the STR locus in the AZF zone of the patient. The karyotype of the patient was 46, XX. SNP array showed a 1.05 Mb 19p12 duplication and a 0.93 Mb Xq27.1 duplication.

CONCLUSIONSThe patient was confirmed as a case of 46,XX male syndrome. The increased copies of the FGF13 gene may be the major causes of abnormal sex determination and testis development.

46, XX Testicular Disorders of Sex Development ; diagnosis ; genetics ; Chromosome Aberrations ; Chromosome Banding ; Genetic Testing ; Humans ; Infertility, Male ; genetics ; Karyotype ; Karyotyping ; Male ; Phenotype ; Sex-Determining Region Y Protein ; genetics

OBJECTIVETo investigate the phenotype-genotype association of isodicentromere Y chromosome by analysis of two female patients carrying the chromosome with sexual development disorders.

METHODSThe karyotypes of the two patients were determined by application of conventional G banding of peripheral blood samples and fluorescence in situ hybridization (FISH). PCR was applied to detect the presence of SRY gene.

RESULTSConventional karyotype analysis showed case 1 to be a mosaic: mos.45,X[38]/46,X,+mar[151]/47,XY,+mar[5]/47,X,+mar × 2[2]/46,XY[4], FISH showed that 12 different cell lines were presented in the karyotype of case 1 and partial cell lines with SRY gene, the marker is an isodicentromere Y chromosome [idic(Y)(p)]. No mutation was found in the SRY gene. The karyotype of case 2 was mos.45,X[25]/46,X,+mar[35]. FISH showed the marker to be an idic(Y)(p) without the SRY gene.

CONCLUSIONThe karyotype of patients carrying idic(Y)(p) seems unstable, and female patients have the characteristics of short stature and secondary sexual hypoplasia. Karyotype analysis combined with FISH analysis can accurately determine the breakpoint of idic(Y) and identify the types of complex mosaic, which may facilitate genetic counseling and prognosis.

Adolescent ; Child ; Chromosomes, Human, Y ; Disorders of Sex Development ; genetics ; Female ; Humans ; Karyotype ; Sex Chromosome Aberrations ; Sex-Determining Region Y Protein ; genetics

The clinical characteristics of patients with disorders of sex development (DSD), and the diagnostic values of classic cytogenetic and molecular genetic assays for DSD were investigated. In the enrolled 56 cases, there were 9 cases of 46,XY DSD, 6 cases of Turner syndrome (TS), one case of Super female syndrome, 25 cases of Klinefelter syndrome, 14 cases of 46,XX DSD, and one case of autosomal balanced rearrangements with hypospadias. The diagnosis of sex was made through physical examination, cytogenetic assay, ultrasonography, gonadal biopsy and hormonal analysis. PCR was used to detect SRY, ZFX, ZFY, DYZ3 and DYZ1 loci on Y and X chromosomes respectively. The DSD patients with the same category had similar clinical characteristics. The karyotypes in peripheral blood lymphocytes of all patients were identified. PCR-based analysis showed presence or absence of the X/Y-linked loci in several cases. Of the 9 cases of 46,XY DSD, 6 were positive for SRY, 9 for ZFX/ZFY, 9 for DYZ3 and 8 for DYZ1 loci. Of the 6 cases of TS, only 1 case with the karyotype of 45,X,/46,XX/46,XY was positive for all 5 loci. Of the 25 cases of Klinefelter syndrome, all were positive for all 5 loci. In one case of rare Klinefelter syndrome variants azoospermia factor (AZF) gene detection revealed the loss of the AZFa+AZFb region. In 14 cases of 46,XX DSD, 7 cases were positive for SRY, 14 for ZFX, 7 for ZFY, 7 for ZYZ3, and 5 for DYZ1. PCR can complement and also confirm cytogenetic studies in the diagnosis of sex in cases of DSD.
Adolescent ; Adult ; Child ; Child, Preschool ; Chromosome Aberrations ; Chromosome Banding ; Chromosomes, Human, X ; genetics ; Chromosomes, Human, Y ; genetics ; Disorders of Sex Development ; diagnosis ; genetics ; Female ; Gene Deletion ; Genetic Loci ; genetics ; Humans ; Karyotyping ; Kruppel-Like Transcription Factors ; genetics ; Male ; Polymerase Chain Reaction ; Sex Chromosome Aberrations ; Sex-Determining Region Y Protein ; genetics ; Young Adult

OBJECTIVETo investigate the value of direct sequencing of sex-determining region Y (SRY) gene, as well as peripheral blood karyotype analysis, in the diagnosis of disorders of sex development (DSD) among children and adolescents with ambiguous genitalia.

METHODSThe karyotypes of 20 children and adolescents with ambiguous genitalia were determined by conventional G-banding analysis. PCR amplification was used to detect SRY gene in these patients, and direct sequencing was used to judge whether there was SRY gene mutation.

RESULTSOf the 20 cases, 17 were positive for SRY gene, and 3 were negative for SRY gene. Direct sequencing revealed no SRY gene mutation in the positive cases, however karyotype analysis found 4 special karyotypes in these patients: 46, XY, del(Y) (q12)/45, X; 46, XY, add(Y) (p11); 46, XY, r(9); 46, XY, 9qh+.

CONCLUSIONSSRY gene detection can help determine the type of DSD among children and has the advantage of quick detection. Used together with G-banding analysis, it is helpful for primary diagnosis of DSD among children.

Adolescent ; Child ; Child, Preschool ; Chromosome Banding ; Disorders of Sex Development ; diagnosis ; genetics ; Humans ; Infant ; Infant, Newborn ; Karyotype ; Sex-Determining Region Y Protein ; genetics

Using Affymetrix's Chicken Genome Array, we used total RNA isolated from the gonads of male and female chicks at embryonic day 9 to identify the genes differentially expressed between male and female. Statistical results show 19 493 genes expressed in male chick's embryonic gonads and 19 368 genes expressed in female. There were 145 genes specificity expressed in male and 189 genes in female. The gene ontology classification (GOC) indicated these differentially expressed genes were mainly involved in cellular component, cell process and molecular banding, a part of genes were involved in organelle component, metabolic process, biologic process, catalytic activity and signal transducer activity. Some genes had reported for sex determination and differentiation in birds, such as avian sex-specific avian sex-specific W-linked, chomodomain-helicase-DNA-binding protein 1 and sex determining region Y-box 9. In addition, we also found several genes or hypothetical proteins were unknown function for the gonad differentiation and development, focus to their biological function and expression pattern in further works would provide a beneficial reference for understand the mechanism of sex differentiation and determination in birds.
Animals ; Chick Embryo ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Gonads ; embryology ; metabolism ; Male ; Oligonucleotide Array Sequence Analysis ; Sex Differentiation ; genetics ; Sex-Determining Region Y Protein ; genetics ; metabolism

Gene expressions are sex-specific in the sex development of mammals. Different genes express in different phases and tend to change with the time. The functions of some genes, such as SRY, SOX9, SOX8, DAX1, and FGF9, have already been defined in male gonadal morphogenesis. This paper presents a review of the genes involved in the formation of the male gonad in mammals.
Animals ; DAX-1 Orphan Nuclear Receptor ; DNA-Binding Proteins ; genetics ; Gene Expression Regulation, Developmental ; Genitalia, Male ; embryology ; growth & development ; metabolism ; High Mobility Group Proteins ; genetics ; Male ; Mammals ; embryology ; genetics ; growth & development ; Morphogenesis ; genetics ; Receptors, Retinoic Acid ; genetics ; Repressor Proteins ; genetics ; SOX9 Transcription Factor ; Sex-Determining Region Y Protein ; genetics ; Transcription Factors ; genetics

OBJECTIVETo test the hypothesis that malignant tumor metastasis is mediated also through a non-cellular, essentially molecular, mechanism in addition to the cellular pathway.

METHODSThe sex-determining region on the Y chromosome was detected as the marker of the primary tumors using PCR in Lewis lung carcinoma (LLC) in vitro and in female C57BL/6 mice bearing LLC with spontaneous metastasis. The macroscopic and microscopic metastases in the tumor-bearing mice were examined for SRY expression by PCR and in situ hybridization, using the tissues from male and female mice as the positive or negative controls.

RESULTS AND CONCLUSIONPositive SRY gene expression was detected in the metastatic foci in the LLC-bearing female mice, suggesting the origination of these tumor cells from the primary tumor foci. We have failed to verify the non-cellular metastasis hypothesis in this animal experiment, but given the limitations of this experiment, we consider further investigation still necessary for verification of this hypothesis using other methods.

Animals ; Carcinoma, Lewis Lung ; genetics ; pathology ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Metastasis ; Neoplasm Transplantation ; Sex-Determining Region Y Protein ; genetics ; metabolism

OBJECTIVETo isolate fetal DNA from maternal plasma and examine its fetal origin.

METHODSFetal DNA in maternal plasma was isolated from 150 samples in the first trimester and mid-trimester of pregnancy, respectively. Real-time fluorescence quantitative polymerase chain reaction PCR (FQ-PCR) was used to determine sex-determining region Y (SRY) gene on Y chromosome.

RESULTSEighty-two women in the first trimester and 90 women in the mid-trimester carried male fetuses,70 and 90 samples of them were positive, respectively. The mean concentrations were (58.82+/-20.90) copies/ml and (152.08+/-62.61) copies/ml. The results of FQ-PCR were negative in the women who carried female fetuses.

CONCLUSIONThe results show that fetal SRY gene can be found at a time as early as 42 days of gestation in maternal plasma by the use of FQ-PCR. The number of fetal DNA increases with gestational age. The real-time FQ-PCR is of great value in the non-invasive prenatal diagnosis.

Adult ; DNA ; blood ; genetics ; isolation & purification ; Female ; Fetus ; metabolism ; Fluorescence ; Gestational Age ; Humans ; Polymerase Chain Reaction ; methods ; Pregnancy ; Pregnancy Trimesters ; Sex-Determining Region Y Protein ; genetics