Klinefelter syndrome (KS) is the most common genetic cause of human male infertility. However, the effect of the extra X chromosome on different testicular cell types remains poorly understood. Here, we profiled testicular single-cell transcriptomes from three KS patients and normal karyotype control individuals. Among the different somatic cells, Sertoli cells showed the greatest transcriptome changes in KS patients. Further analysis showed that X-inactive-specific transcript ( XIST ), a key factor that inactivates one X chromosome in female mammals, was widely expressed in each testicular somatic cell type but not in Sertoli cells. The loss of XIST in Sertoli cells leads to an increased level of X chromosome genes, and further disrupts their transcription pattern and cellular function. This phenomenon was not detected in other somatic cells such as Leydig cells and vascular endothelial cells. These results proposed a new mechanism to explain why testicular atrophy in KS patients is heterogeneous with loss of seminiferous tubules but interstitial hyperplasia. Our study provides a theoretical basis for subsequent research and related treatment of KS by identifying Sertoli cell-specific X chromosome inactivation failure.
Animals ; Humans ; Male ; Female ; Sertoli Cells/metabolism* ; Klinefelter Syndrome/genetics* ; Endothelial Cells ; Testis/metabolism* ; X Chromosome/metabolism* ; Mammals/genetics*

OBJECTIVETo analyze the numerical aberration of chromosome X, Y and 18 in the spermatozoa of asthenospermia patients by triple-color fluorescence in situ hybridization.

METHODSThe experiment included 10 asthenospermia patients and 5 healthy men with normal semen quality as controls. Fluorescence in situ hybridization (FISH) and probes for chromosomes including X, Y and 18 were used to determine the frequency of the aneuploid of the chromosomes in spermatozoa.

RESULTSOf the 45,547 spermatozoa counted from the semen samples, the hybridization rate was 99.18%. The frequencies of the chromosome disomies including XX18, XY18, YY18, X1818 and Y1818 were (0.124 +/- -0.086)%, (0.360 +/- 0.380)%, (0.109 +/- 0.195)%, (0.342 +/- 0.746)% and (0.299 +/- 0.564)% in the case group and (0.014 +/- 0.019)%, (0.090 +/- 0.080)%, (0.030 +/- 0.031)%, (0.068 +/- 0.103)% and (0.075 +/- 0.083)% in the control. The sperm aneuploid rate was 9.25% in the former and 2.70% in the latter, with significant difference in between (P< 0.01).

CONCLUSIONAsthenospermia patients have a higher aneuploid rate of sperm chromosome than normal fertile men. However, larger samples are yet to be studied to obtain more scientific evidence.

Aneuploidy ; Asthenozoospermia ; genetics ; Chromosome Painting ; methods ; Chromosomes, Human, Pair 18 ; Chromosomes, Human, X ; Chromosomes, Human, Y ; Humans ; Male ; Sex Chromosome Aberrations ; Spermatozoa ; metabolism

OBJECTIVETo analyze a fetus with increased nuchal translucency and nuchal fold, and to assess the recurrence risk for her family and provide a basis for prenatal diagnosis.

METHODSG-banded karyotyping and single nucleotide polymorphism-based array (SNP-Array) analysis were used to analyze the fetus and her parents.

RESULTSSNP-Array analysis has detected a 41.04 Mb duplication at Xp22.33p11.4 and a 30.51 Mb duplication at 13q31.3q34 in the fetus. G-banding karyotyping indicated that the fetus had a karyotype of 46,X,der(X)(13qter-13q31::Xp11.4-Xp22.3::Xp22.3-Xqter). Her parents had normal results for both G-banding karyotyping and SNP-Array analysis, suggesting that the fetus has carried a de novo derivative chromosome X.

CONCLUSIONSNP-Array combined with G-banding karyotyping is helpful to confirm the composition and connection type of de novo derivative chromosome, which can improve the accuracy of diagnosis and is valuable for the evaluation of recurrence risk.

Adult ; Chromosome Banding ; Chromosome Duplication ; Chromosomes, Human, X ; genetics ; Female ; Fetus ; abnormalities ; metabolism ; Humans ; Karyotyping ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Polymorphism, Single Nucleotide ; Pregnancy ; Prenatal Diagnosis ; methods ; Sex Chromosome Aberrations

OBJECTIVETo study the clonality of palmar fibromatosis by molecular genetic analysis of X chromosome inactivation pattern at a polymorphic site of human androgen receptor gene (HUMARA).

METHODSTwelve female cases of palmar fibromatosis were enrolled into this study. Hematoxylin and eosin-stained sections of paraffin-embedded, formalin-fixed tissues were microdissected by laser capture microdissection technology in order to obtain the proliferative spindle cells. Tumor cells isolated from rectal adenocarcinoma in a female patient were used as positive control. The genomic DNAs were extracted with phenol and chloroform, digested with methylation-sensitive restriction endonuclease HpaII, and amplified by polymerase chain reaction (PCR) using primers targeted to a highly polymorphic short tandem repeat of HUMARA. The amplimers were separated on vertical 8% non-denaturing polyacrylamide gels and the patterns were visualized with ethidium bromide stain.

RESULTSThe methodology for clonality analysis was validated in the positive control using rectal adenocarcinoma cells. Among the 12 cases studied, PCR amplification failed in 3 samples and 1 sample showed homozygosity which was not suitable for further analysis. Eight samples were successfully amplified and showed a random X chromosome inactivation pattern, suggesting polyclonality in these lesions.

CONCLUSIONSPalmar fibromatosis is a polyclonal condition and should be considered as a form of non-neoplasmic fibroblastic proliferation.

Actins ; metabolism ; Adolescent ; Adult ; Aged ; Child ; Chromosomes, Human, X ; Clone Cells ; metabolism ; pathology ; Dupuytren Contracture ; genetics ; metabolism ; pathology ; Female ; Humans ; Middle Aged ; Polymerase Chain Reaction ; Receptors, Androgen ; genetics ; metabolism ; X Chromosome Inactivation

OBJECTIVETo investigate the constitution of abnormal spermatozoa from patients with sex chromosome anomalies.

METHODSTriple color fluorescence in situ hybridization (FISH) was used to determine the sex chromosome constitution of spermatozoa from three patients with sex chromosome anomalies (case 1:46,XY/47,XXY, case 2:45,XO/46,X,Yqh-, case 3:47,XXY). The preimplantation genetic diagnosis (PGD) was performed to case 2.

RESULTSAn increased ratio (2.05 vs 1) of X-bearing to Y-bearing spermatozoa was only observed in case 2, who also had an increased incidence of total abnormal spermatozoa (29.71%). An increased incidence of total abnormal spermatozoa (4.91%) was also observed in case 3. Among the constitution of abnormal spermatozoa, case 2 had the increased proportions of XY18 disomy, O18 monosomy and XO monosomy, while case 3 had an increase proportion of XY18 disomy (1.87%). PGD was performed to case 2 and one embryo with XX1818 was selected for implanting.

CONCLUSIONUsing FISH to detect the sperm sex chromosomes in patients with sex chromosome anomalies can provide the useful information to evaluate the risk of sex chromosome anomalies in preimplantation embryos.

Adult ; Chromosomes, Human, X ; genetics ; Chromosomes, Human, Y ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Preimplantation Diagnosis ; methods ; Sex Chromosome Aberrations ; Spermatozoa ; metabolism

OBJECTIVETo observe the relationship between skewed X-chromosomal inactivation (SXCI) and development of lung cancer in females.

METHODSDNA was isolated from peripheral blood cells from patients with primary lung cancer (n = 148) and control subjects (n =289). Exon 1 of androgen receptor ( AR) gene was amplified, with its products from different alleles resolved on denaturing polyacrylamide gels and visualized by silver staining. The corrected ratio (CR) between products from different AR alleles before and after Hpa II pretreatment was calculated. All statistical tests were two-sided.

RESULTSWith CR> or = 10 adopted as the criterion, SXCI was found more frequently in the younger patients ( C50 years; 7. 9%) than in the controls of the same age group (1. 2% ; P = 0. 046). The SXCI frequency, however, were not significantly different between the old patients ( > 50 years; 4. 5% ) and the controls of the same age group (5. 4% ; P =0. 488). Whether taking CR> or =3 or CR> or =10 as the criteria, the average ages of the patients with SXCI were more than 10 years younger than those without SXCI (P < 0. 05).

CONCLUSIONSXCI in blood cells is associated with early development of lung cancer in females.

Adult ; Age Factors ; Aged ; Alleles ; Chromosomes, Human, X ; genetics ; DNA ; genetics ; metabolism ; Deoxyribonuclease HpaII ; metabolism ; Exons ; Female ; Genetic Predisposition to Disease ; Humans ; Lung Neoplasms ; blood ; genetics ; pathology ; Middle Aged ; Polymerase Chain Reaction ; Receptors, Androgen ; genetics ; X Chromosome Inactivation

In recent years, large numbers of non-coding RNAs (ncRNAs) have been identified in C. elegans but their functions are still not well studied. In C. elegans, CEP-1 is the sole homolog of the p53 family of genes. In order to obtain transcription profiles of ncRNAs regulated by CEP-1 under normal and UV stressed conditions, we applied the 'not-so-random' hexamers priming strategy to RNA sequencing in C. elegans, This NSR-seq strategy efficiently depleted rRNA transcripts from the samples and showed high technical replicability. We identified more than 1,000 ncRNAs whose apparent expression was repressed by CEP-1, while around 200 were activated. Around 40% of the CEP-1 activated ncRNAs promoters contain a putative CEP-1-binding site. CEP-1 regulated ncRNAs were frequently clustered and concentrated on the X chromosome. These results indicate that numerous ncRNAs are involved in CEP-1 transcriptional network and that these are especially enriched on the X chromosome in C. elegans.
Animals ; Binding Sites ; Caenorhabditis elegans ; Caenorhabditis elegans Proteins ; genetics ; metabolism ; High-Throughput Nucleotide Sequencing ; Promoter Regions, Genetic ; RNA, Untranslated ; metabolism ; Sequence Analysis, RNA ; Transcriptome ; radiation effects ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Ultraviolet Rays ; X Chromosome

OBJECTIVETo investigate the pathogenesis of male sterility in 48,XXYY syndrome patient.

METHODSThe peripheral lymphocyte was detected by dual-color fluorescence in situ hybridization. The bioptic testicular tissues were pathologically sectioned and ultra-thin sections were examined by electron-microscopy.

RESULTSThe pathological findings revealed extremely severe dysgenesis of the badly damaged testicular tissue. Only a few convoluted seminiferous tubules were found, in which no spermatogenic cell or sperm of any range could be viewed. The ultrastructural observations showed the thickened interstitial vascular walls of the testicular tissue and severe hyperplasia of the collagen fibers in the basilemma and lumens of the blood vessels.

CONCLUSIONThe structure of the testicle in the 48,XXYY syndrome patient has severe fibrous hyperplasia, leading to the non-specific thickening of the barrier and serious damage to the blood-testis barrier, which in turn produce significant disturbance and pathological changes in the process of the spermatogenic cell formation. The whole interrelated loops account mainly for the male sterility.

Adult ; Chromosomes, Human, X ; genetics ; Chromosomes, Human, Y ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infertility, Male ; genetics ; pathology ; Lymphocytes ; metabolism ; Male ; Microscopy, Electron ; Sex Chromosome Aberrations ; Testis ; metabolism ; pathology ; ultrastructure

The study was aimed to investigate the immunophenotypic and cytogenetic features of chronic lymphocytic leukemia (CLL) in order to provide an evidence for diagnosis and therapy. Immunophenotypic analysis was performed by using a panel of monoclonal antibodies and three-color immunofluorescence staining methods of flow cytometry in 51 patients with CLL, and the cytogenetic features were analyzed by R-banding technique. The results indicated that among 51 CLL cases, the positive rate of CD19 and CD23 was 96.1%, followed by CD15 (94.1%), CD20 (82.4%) and CD22 (78.4%). The positive rate of CD38 was 23.5%. Forty-six patients expressed both CD5 and CD19. Seven main clonal chromosomal abnormalities were detected by conventional cytogenetics (CC) in eighteen cases (35.3%), with three cases of +12, two cases of 13q(-), other chromosomal abnormalities included +14, 6q(-), t (11; 14), t (14; 18) and t (2; 7). Expression of the antigens had no relationship with chromosomal abnormalities. It is concluded that typical CLL express CD5, CD19 and CD23, and the positive rate detected by CC in CLL is low. Immunophenotyping in combination with cytogenetic technique plays an important role in the diagnosis and prognosis of CLL.
Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; Antigens, CD ; metabolism ; Antigens, CD19 ; metabolism ; Antigens, CD20 ; metabolism ; Chromosome Aberrations ; Chromosomes, Human, Pair 12 ; Chromosomes, Human, Pair 13 ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; immunology ; Lewis X Antigen ; metabolism ; Male ; Middle Aged ; Translocation, Genetic ; genetics

X-linked severe combined immunodeficiency (X-SCID) is a rare, life-threatening immune disorder, caused by mutations in the gamma c chain gene, which encodes an essential component of the cytokine receptors for interleukin-2 (IL-2), IL-4, IL-7, IL-9, IL-15, and IL-21. A 13-month-old boy with recurrent infections who had reduced serum immunoglobulin levels and decreased numbers of CD3, CD16/56 cells was evaluated for gamma c chain gene mutation and protein expression. The patient had a C-to-T point mutation at nucleotide position 690, one of the hot spots, resulting in a single amino acid substitution of cysteine for arginine (R226C), as determined by direct sequencing and PCR-RFLP. The patient's mother was a heterozygous carrier. Percutaneous umbilical cord blood sampling was performed at the 6-month of gestation in a subsequent pregnancy. As the immunophenotype of the fetus showed an identical pattern, the pregnancy was terminated and genetic analysis of the abortus confirmed recurrence. This is the first report of the molecular diagnosis of X-SCID in Korea. Genetic analysis of the gamma c chain gene is useful for definite diagnosis and genetic counseling for X-SCID.
Arginine/chemistry ; Cysteine/chemistry ; DNA/metabolism ; DNA Mutational Analysis ; Female ; Flow Cytometry ; Genetic Counseling/*methods ; Heterozygote ; Human ; Immunoglobulins/metabolism ; Immunophenotyping/methods ; Korea ; *Linkage (Genetics) ; Male ; *Mutation ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Receptors, Immunologic/*genetics ; Sequence Analysis, DNA ; Severe Combined Immunodeficiency/*diagnosis/*genetics ; Support, Non-U.S. Gov't ; Time Factors ; *X Chromosome