A 15-year-old boy was referred due to gynecomastia and short stature. He was overweight and showed the knuckle-dimple sign on the left hand, a short fourth toe on the left foot, and male external genitalia with a small phallus. His levels of estradiol and follicle-stimulating hormone were increased, and his testosterone concentration was normal. Other hormonal tests were within the normal range. Radiographs showed short fourth and fifth metacarpals and fourth metatarsal bones. The karyotype was reported as 46,X,+mar, and the marker chromosome was shown to originate from the Y chromosome, which was identified by fluorescence in situ hybridization. Polymerase chain reaction and direct sequencing were used to clarify the deleted loci of the Y chromosome by making use of Y-specific sequence-tagged sites (STSs). The sex-determining region Y and centromere were verified, and there were microdeletions on the long arm of the Y chromosome. The azoospermia factor (AZF) b region was partially deleted, and AZFa and AZFc were completely deleted. Two STS probes of sY143 and the Y chromosome RNA recognition motif in AZFb showed positive signals corresponding to Yq11.223. The karyotype of the patient was interpreted as 46,X,der(Y)del(Y)(q11.21q11.222)del(Y)(q11.23qter). Herein, we report a rare case of a boy presenting with gynecomastia and short stature with 46, X, +mar, which originated from the Y chromosome, which was identified to have Yq microdeletions.
Adolescent ; Arm ; Azoospermia ; Centromere ; Estradiol ; Fluorescence ; Follicle Stimulating Hormone ; Foot ; Genitalia ; Gynecomastia* ; Hand ; Humans ; In Situ Hybridization ; Karyotype ; Male* ; Metacarpal Bones ; Metatarsal Bones ; Overweight ; Polymerase Chain Reaction ; Reference Values ; RNA ; Sequence Tagged Sites ; Testosterone ; Toes ; Y Chromosome

Interchromosomal insertion of Y chromosome heterochromatin in an autosome was identified in a fetus and a family. A fetal karyotype was analyzed as 46,XX,dup(7)(?q22q21.1) in a referred amniocentesis at 16 weeks of gestation for advanced maternal age. In the familial karyotype analyses for identification of der(7), the mother, the first daughter and the maternal grandmother showed the same der(7) as the fetus's. CBG-banding was positive at 7q22 region of der(7) that indicated inserted material was originated from heterochromatin. The origin of heterochromatic insertion region in der(7) of the fetus and the mother was found in Yq12 region by fluorescent in situ hybridization with a DYZ1 probe. In the specific analysis of Y chromosomal heterochromatic region of ins(7;Y) of the mother, 15 sequence tagged sites from Yp11.3 region including SRY to Yq11.223 region was not detected. Final karyotypes of the mother, the first daughter and the maternal grandmother were reported as 46,XX,der(7)ins(7;Y)(q21.3;q12q12). All female carriers of ins(7;Y) in the family showed normal phenotype and the mother and the maternal grandmother were fertile. A healthy girl was born at term. We report a rare case of familial interchromosomal insertion of Y chromosome heterochromatin detected only in female family members with normal phenotype that was diagnosed prenatally.
Amniocentesis ; Female ; Fetus ; Grandparents ; Heterochromatin* ; Humans ; In Situ Hybridization, Fluorescence ; Karyotype ; Maternal Age ; Mothers ; Nuclear Family ; Phenotype ; Pregnancy ; Prenatal Diagnosis* ; Sequence Tagged Sites ; Y Chromosome*

OBJECTIVETo investigate the possibility of applying multiplex ligation-dependent probe amplification (MLPA) to the detection of azoospermia factor (AZF) microdeletion on the Y chromosome in infertile men with azoospermia or severe oligozoospermia.

METHODSDNA samples were obtained from 147 azoospermia or severe oligozoospermia patients and 154 normal controls. After denatured at 95 degrees C, the samples were hybridized to the specific probes designed for the AZF region. With the ligase, the hybrid products were amplified by a pair of universal primers labeled with FAM fluorescence, and then separated by capillary electrophoresis for data analysis. Meanwhile all the samples were subjected to multiplex-PCR (mPCR) analysis for sequence-tagged sites (STS) in the AZF region.

RESULTSSTS deletion was detected in 22 (15.0%) of the 147 patients but not in the normal controls. By MLPA, 40 (27.2%) of the patients were found with specific probe omission in the AZF region, as compared with 20 cases in the control group.

CONCLUSIONCompared with mPCR, MLPA has a better sensitivity in detecting AZF microdeletions, and it provides more precise genetic information on the AZF regions, which may contribute to in-depth exploration into the etiological mechanism of impaired spermatogenesis.

Adult ; Azoospermia ; genetics ; Case-Control Studies ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; DNA Probes ; Genetic Loci ; Humans ; Infertility, Male ; Male ; Nucleic Acid Amplification Techniques ; methods ; Oligospermia ; genetics ; Polymerase Chain Reaction ; methods ; Seminal Plasma Proteins ; genetics ; Sequence Tagged Sites ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; genetics ; Young Adult

OBJECTIVETo observe peripheral blood chromosome abnormality and microdeletions of the SRY and AZF genes on the Y chromosome in patients with chimera Klinefelter syndrome.

METHODSWe analyzed the cytogenetic karyotype of the peripheral blood chromosome in 1 infertile patient with mosaic karyotype Klinefelter syndrome and his parents. We identified 9 sequence tagged sites (STS) by multiplex PCR: sY84, sY86, sY127, sY129, sY134, sY254, sY255, sY242, and sY152. Meanwhile we detected the SRYgene and the microdeletion of AZF using ZFX/ZFY as the internal control gene.

RESULTSThe karyotype of the patient was 46,XY (12%)/47,XXY (30%)/48,XXYY (56%)/49,XXXXY (2%). The karyotypes of his parents were normal. Consistency was found between the SRY gene and the chromosome gender in the patient and his parents. Y chromosome AZF microdeletion was observed in the patient. The deletion sites were sY86 and sY127, and the deletion type was AZFa + AZFb.

CONCLUSIONAZF microdeletion of the Y chromosome exists in patients with Klinefelter syndrome. Chromosome karyotype and Y-chromosome AZF microdeletion are important criteria for the genetic diagnosis of Klinefelter syndrome.

Chimerism ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Cytogenetics ; Humans ; Infertility, Male ; Karyotyping ; Klinefelter Syndrome ; genetics ; Male ; Sequence Tagged Sites ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; genetics ; Young Adult

OBJECTIVETo investigate the breakpoints of the azoospermia factor c (AZFc) microdeletion in Chinese Han population.

METHODSWe detected 9 sequence tagged sites (sY84, sY86, sY127, sY134, sY152, sY145, sY255, sY254 and sY157) to confirm AZFc microdeletions in the Y chromosome for patients with severe oligozoospermia or non-obstructive azoospermia by multiplex polymerase reaction. To locate the breakpoints of AZFc microdeletions, we analyzed 192 patients with sY255, sY254 and sY157 dele- ted by detecting sY1191, sY1197, sY1054, sY1125 and sY1206, respectively.

RESULTSFive breaking patterns were found in the 192 patients with sY255, sY254 and sY157 deleted, among which the common ones were sY1197(+), sY1191(-), sY1054(-), sY1206(-) and sY1125(+), which accounted fro 54.17% (104/192), sY1197(+), sY1191(+), sY1206(-), sY1054(-) and sY1125(+), which constituted 28.12% (54/192), sY1197(+), sY1191(-), sY1206(-), sY1054(+) and sY1125 (+), which made up 14.58% (28/192). The proximal breakpoint located between sY1197 and sY1191 was 70.83% of AZFc microdeletions, and the distant breakpoint located between sY1054 and sY1125 was 82.29%.

CONCLUSIONThere are 5 breaking patterns of AZFc microdeletions in Chinese Han population, the proximal and distant breakpoints mostly located at the replicons b2 and b4, respectively.

Asian Continental Ancestry Group ; genetics ; Azoospermia ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; Humans ; Infertility, Male ; genetics ; Male ; Sequence Tagged Sites

OBJECTIVETo investigate the patterns of azoospermia factor C (AZFc) microdeletion in the Chinese Han population and optimize the selection of the required sequence tagged sites (STSs) of AZF microdeletion in multiplex polymerase chain reaction (PCR).

METHODSNine STSs (sY84, sY86, sY127, sY134, sY152, sY145, sY255, sY254 and sY157) were detected by multiplex polymerase reaction for Y chromosome microdeletion in 164 Chinese Han patients with severe oligozoospermia or non-obstructive azoospermia, and another 105 with normal sperm concentration were included as controls. Meanwhile 180 cases of AZFc microdeletion (absence of sY255 and sY254) from multiple reproductive medical centers were analyzed for sY145, sY152 and sY157.

RESULTSFourteen (8.5%) of the 164 patients with severe oligozoospermia or non-obstructive azoospermia showed AZFc microdeletion (absence of sY255 and sY254). All the 194 patients with the absence of sY255 and sY254 displayed the presence of sY145 and sY152, only 2 of them with sY157 present. Deletion of sY1206 and DAZ3/DAZ4 copies was confirmed in 1 case of severe oligozoospermia with sY157 absent only.

CONCLUSIONDeletion of sY255 and sY254 as well as sY157 is the most common pattern of AZFc microdeletion in the Chinese Han population. sY145 and sY152 can be omitted in AZFc screening. Absence of sY157 alone may be a new type of partial AZFc microdeletion in the Chinese Han population, and the clinical significance of unique sY157 absent needs to be further explored.

Asian Continental Ancestry Group ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; Genetic Loci ; genetics ; Humans ; Infertility, Male ; Male ; Oligospermia ; genetics ; Polymerase Chain Reaction ; Sequence Tagged Sites ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development

OBJECTIVETo screen and determine the regions of copy number variation (CNV) associated with hepatocellular carcinoma (HCC) using SNP array and fluorescence quantitative PCR.

METHODSThe CNV from HCC cell line TJ3ZX-01 was analyzed using GeneChip Human Mapping 500K SNP array. According to the data obtained by SNP array analysis, four candidate amplification regions were verified in 41 primary HCC samples by fluorescence quantitative PCR.

RESULTSFour regions of copy number amplification at 1q21.2, 1q22 approximately 23.1, 7p22.1 and 22q13.1 were detected by SNP array analysis. The four candidate amplicons occurred in 56.1% (23/41) of HCC samples at 1q21.2; 80.5% (33/41) at 1q22 approximately 23.1; 75.6% (31/41) at 7p22.1 and 31.7% (13/41) at 22q13.1 analyzed with sequence tagged site (STS) markers by quantitative PCR.

CONCLUSIONIn four candidate amplification regions selected by SNP array analysis and detected by fluorescence quantitative PCR, three amplification regions show increased copy number in more than 50.0% HCC tissues. This result indicates that these amplification regions are associated with pathogenesis of hepatocellular carcinoma.

Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; Chromosomes, Human, Pair 1 ; genetics ; Chromosomes, Human, Pair 22 ; genetics ; Chromosomes, Human, Pair 7 ; genetics ; DNA Copy Number Variations ; genetics ; Female ; Humans ; Liver Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; Sequence Tagged Sites

OBJECTIVETo establish a liquid chip technology to detect Y chromosome microdeletions in Chinese infertile males with azoospermia or oligozoospermia.

METHODSMultiplex PCR and liquid chip technology were used to detect the Y chromosome microdeletions in AZF region in 178 infertile patients with azoospermia and 134 infertile patients with oligozoospermia as well as 40 fertile control men.

RESULTSForty out of 312 patients (12.8%) were found to have deletions in AZF region. The microdeletion frequency was 14%(25/178) in the azoospermic group, 9.6%(11/114) in the oligospermic and 20%(4/20) in the severe oligospermic group.

CONCLUSIONThe authors developed a high-throughput, fast and simple assay to screen the AZF region microdeletions of Y chromosome.

Asian Continental Ancestry Group ; genetics ; Azoospermia ; genetics ; Base Sequence ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Electrophoresis, Polyacrylamide Gel ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Oligospermia ; genetics ; Sequence Tagged Sites

OBJECTIVEWe previously showed that introduction of a normal, neomycin-tagged human chromosome 8 reduced the metastatic capacity of C5F rat liver cancer cell line, which had high metastatic potential without affecting tumorigenicity, suggesting the presence of one or more metastasis suppressor genes encoded on human chromosome 8. We proceeded to define further the region harboring the metastasis suppressor gene(s) and to determine the random loss of human chromosome 8 by PCR amplification of sequence tag site (STS) markers.

METHODSThe national Center for Biotechnology Information (NCBI) databases were used as references of the relative genetic distances of the STS markers. C5F genomic DNA and A9/neo8 genomic DNA were used as negative and positive controls for chromosome 8 amplification, respectively. Genomic DNA was isolated and quantified from cultured hybrid clones (A9/C5F-1 and A9/C5F-2 microcell hybrid clones served as metastasis-unsuppressed groups; A9/C5F-4, A9/C5F-8 and A9/C5F-10 microcell hybrid clones served as metastasis suppressed groups). STS-PCR products were separated by electrophoresis through 2% agarose gel.

RESULTSMetastasis-suppressed microcell hybrid clones (A9/C5F-4, A9/C5F-8 and A9/C5F-10) conserved STS markers between D8S542 --> D8S1973 (8p21.1-23.1). In contrast, metastasis-unsuppressed clones (A9/C5F-1 and A9/C5F-2) lacked several markers in this region. In attempts to refine the region retained in the microcell suppressed clones, more densely spaced STS markers in the human chromosome 8p21.1-23.1 were used. We found that the metastasis-suppressed clones retained 18cM region between D8S542 and D8S1973 (8P21.1-23.1), where as the metastasis-unsuppressed clones lacked the region.

CONCLUSIONOur results suggest that a metastasis suppressor gene is located within the interval between D8S542 and D8S1973 on human chromosome 8p21.1-23.1.

Carcinoma, Hepatocellular ; genetics ; Cell Line ; Cell Line, Tumor ; Chromosome Mapping ; Chromosomes, Human, Pair 8 ; genetics ; Fibroblasts ; cytology ; Genes, Tumor Suppressor ; Humans ; In Situ Hybridization, Fluorescence ; Liver Neoplasms ; genetics ; Neoplasm Metastasis ; Sequence Tagged Sites

OBJECTIVESearching a new molecular method to authenticate Panax ginseng and P. quenquefolium.

METHODSingle primers based on rDNA sequences of Panax species were designed to obtain polymorphic bands of P. ginseng and P. quinquefolius and then sequenced. Four PCR primers (two forword and two reverse primers) specific to P. ginseng and P. quinquefolius were designed.

RESULTPrimer Pg-6F, Pg-479R only amplified 474 bp band for P. ginseng and primer Pq-442F, Pq-658R only amplified 217 bp band for P. quinquefolius. It is indicated that the four primers could serve as specific STS primers for Panax species.

CONCLUSIONA new way to obtain STS primers of Panax species was established. This method is more quick and efficient than SCAR-PCR method and can serve as a model to obtain molecular markers for other Chinese material medica.

Base Sequence ; DNA Primers ; DNA, Plant ; genetics ; DNA, Ribosomal ; genetics ; Genetic Markers ; genetics ; Molecular Sequence Data ; Panax ; classification ; genetics ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; Polymorphism, Genetic ; Random Amplified Polymorphic DNA Technique ; methods ; Sequence Analysis, DNA ; Sequence Tagged Sites ; Species Specificity