Y chromosome structural variation in infertile men detected by targeted next-generation sequencing.

PURPOSE: To provide a validated method to identify copy number variation (CNV) in regions of the Y chromosome of infertile men by next-generation sequencing (NGS). METHODS: Semen analysis was used to determine the quality of semen and diagnose infertility. Deletion of the azoospermia factor (AZF) region in the Y chromosome was detected by a routine sequence-tagged-site PCR (STS-PCR) method. We then used the NGS method to detect CNV in the AZF region, including deletions and duplications. RESULTS: A total of 326 samples from male infertility patients, family members, and sperm donors were studied between January 2011 and May 2017. AZF microdeletions were detected in 120 patients by STS-PCR, and these results were consistent with the results from NGS. In addition, of the 160 patients and male family members who had no microdeletions detected by STS-PCR, 51 cases were found to exhibit Y chromosome structural variations by the NGS method (31.88%, 51/160). No microdeletions were found in 46 donors by STS-PCR, but the NGS method revealed 11 of these donors (23.91%, 11/46) carried structural variations, which were mainly in the AZFc region, including partial deletions and duplications. CONCLUSION: The established NGS method can replace the conventional STS-PCR method to detect Y chromosome microdeletions. The NGS method can detect CNV, such as partial deletion or duplication, and provide details of the abnormal range and size of variations.

Associated factors of secondary sex ratio of offspring in assisted reproductive technology: a cross-sectional study in Jilin Province, China.

BACKGROUND: The aim of this study was to determine the secondary sex ratio (SSR) of offspring in assisted reproduction technology (ART) in Jilin Province, China, and to analyse the influencing factors associated with SSR. METHODS: A cross-sectional study of 3833 babies including singletons and twins born to 2990 couples treated by ART between May 2011 and December 2018 was performed. RESULTS: The main outcomes of this study were that the SSR of ART babies in Jilin Province was 50.64% and the SSR was associated with fertilization methods (p < 0.05). Comparing to in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) (OR = 0.808, 95%CI: 0.681-0.958) decreased the percentage of male babies. CONCLUSIONS: This study suggests that the SSR of ART births in Jilin Province was lower than the normal level and ICSI had a significant effect on SSR. Though we need more samples to study in the future, we still need to think about the impact of ICSI on SSR in ART.

Identification of KISS1R gene mutations in disorders of non-obstructive azoospermia in the northeast population of China.

BACKGROUND: Non-obstructive azoospermia (NOA), a serious phenotype of male spermatogenesis failure, is a multifactorial disease which is regulated by genetic, epigenetic, and environmental factors. Some gene structural variants have been demonstrated to be related to NOA. Loss-of-function mutations of KISS1R cause normosmic idiopathic hypogonadotropic hypogonadism (IHH) which result in azoospermia at the pre-testicular level. The objective of this research was to investigate genetic variants of KISS1R in NOA patients. METHODS: The entire coding region of 52 spermatogenesis-associated genes (KISS1R included) was sequenced from 200 NOA patients. Mutation screening was performed to identify genetic variations of these genes by targeted exome sequencing. Sequencing data analysis was carried out by a series of bioinformatics tools. Candidate variants confirmation was performed by Sanger sequencing. Functional analysis of candidate variants was evaluated using SIFT and PolyPhen-2. RESULTS: Three heterozygous missense variants in KISS1R were identified in three patients, respectively. No deleterious variations in other candidate genes were found in the three patients. Two of these three variants, p.A211T and p.G186E, had been reported in the ExAC and dbSNP database, respectively, while the other variant p.A301D was novel. These variants were all predicted to be likely pathogenic by in silico analysis. CONCLUSION: Our study revealed three heterozygous missense variants in KISS1R which expanded the mutation spectrum of KISS1R in infertile men with NOA in the northeast of China.

Targeted Next-Generation Sequencing Identifies Novel Sequence Variations of Genes Associated with Nonobstructive Azoospermia in the Han Population of Northeast China.

BACKGROUND This study aimed to screen common and low-frequency variants of nonobstructive azoospermia (NOA)-associated genes, and to construct a database for NOA-associated single nucleotide variants (SNVs). MATERIAL AND METHODS Next-generation sequencing of 466 NOA-associated genes was performed in 34 patients with NOA (mean age, 29.06±4.49 years) and 40 sperm donors (mean age, 25.08±5.75 years) from the Han population of northeast China. The SNV database was constructed by summarizing NOA non-negatively-associated SNVs showing statistical differences between NOA cases and controls, and then selecting low-frequency variants using Baylor's pipeline, to identify statistically valid SNVs. RESULTS There were 65 SNVs identified that were significantly different between both groups (p<0.05). Five genetic variants showed positive correlations with NOA: MTRR c.537T>C (rs161870), odds ratios (OR), 3.686, 95% confidence interval (CI), 1.228-11.066; MTRR, c.1049A>G (rs162036), OR, 3.686, 95% CI, 1.228-11.066; PIWIL1, c.1580G>A (rs1106042), OR, 4.737, 95% CI, 1.314-17.072; TAF4B, c.1815T>C (rs1677016), OR, 3.599, 95% CI, 1.255-10.327; and SOX10 c.927T>C (rs139884), OR, 3.192, 95% CI, 1.220-8.353. Also, 52 NOA non-negatively associated SNVs and 39 SNVs were identified by Baylor's pipeline and selected for the SNV database. CONCLUSIONS Five genetic variants were shown to have positive correlations with NOA. The SNV database constructed contained NOA non-negatively associated SNVs and low-frequency variants. This study showed that this approach was an effective strategy to identify risk alleles of NOA.

Fluorescent 6-amino-6-deoxyglycoconjugates for glucose transporter mediated bioimaging.

Two novel fluorescent bioprobes, namely, 6N-Gly-Cy3 and 6N-Gly-Cy5, were designed and synthesized for real-time glucose transport imaging as well as potentially useful tracer for galactokinase metabolism. The structure of the bioprobes was fully characterized by 1H NMR, 13C NMR, IR, and HRMS. The fluorescence properties, glucose transporter (GLUT) specificity, and the quenching and safety profiles were studied. The cellular uptake of both bioprobes was competitively diminished by d-glucose, 2-deoxy-d-glucose and GLUT specific inhibitor in a dose-dependent manner in human colon cancer cells (HT29). Comparison study results revealed that the 6N-derived bioprobes are more useful for real-time imaging of cell-based glucose uptake than the structurally similar fluorescent tracer 6-NBDG which was not applicable under physiological conditions. The up to 96 h long-lasting quenching property of 6N-Gly-Cy5 in HT29 suggested the potential applcability of the probe for cell labeling in xenograft transplantation as well as in vivo animal imaging studies.

Placental mosaicism for Trisomy 13: a challenge in providing the cell-free fetal DNA testing.

PURPOSE: We investigated the disagreement between the positive cell-free fetal DNA test for trisomy 13 and the standard cytogenetic diagnosis of one case. METHODS: Cell-free fetal DNA testing was performed by massively parallel sequencing. We used conventional cytogenetic analysis to confirm the commercial cell-free fetal DNA testing. Additionally, postnatal fluorescent in situ hybridization (FISH) testing was performed on placental tissues. RESULTS: The cell-free fetal DNA testing result was positive for trisomy 13. G-banded analysis of amniotic fluid was normal, 46, XY. FISH testing of tissues from four quadrants of the placenta demonstrated mosaicism for trisomy 13. CONCLUSIONS: A positive cell-free fetal DNA testing result may not be representative of the fetal karyotype because of placental mosaicism. Cytogenetic analysis should be performed when abnormal cell-free fetal DNA test results are obtained.

[Predictive value of abnormal second-trimester maternal serum triple screening markers for adverse pregnancy outcomes].

OBJECTIVE: To investigate the predictive value of abnormal multiples of the median (MoM) of second trimester maternal serum triple screening (STMSTS) markers for adverse pregnancy outcomes. METHODS: 16 000 singleton pregnancies at 15⁺° to 20⁺5 weeks' gestation who underwent STMSTS between July 2010 and January 2013 in the First Hospital of Jilin University were recruited. Maternal serum AFP, free ß-hCG (F-ß-hCG) and unconjugated estriol (uE3) levels were measured using time- resolved fluoroimmunoassay, and then converted to MoM. LifeCycle 3.2 software was used to calculate risk, and a risk value greater than 1 in 270 or 1 in 350 was considered as high risk for trisomy 21 syndrome (Down syndrome, DS) and trisomy 18 syndrome (Edwards syndrome, ES), respectively. MoM of AFP more than 2.5 was considered high risk for open neural tube defect (ONTD). Amniocentesis and karyotyping, ultrasound screening were advised for high risk women. AFP, F-ß-hCG higher than 2.0 MoM or uE3 lower than 0.5 MoM was considered as abnormal, respectively. The MoM of STMSTS marker between women with adverse pregnancy outcome and with normal outcome was compared. RESULTS: (1) The median MoM of AFP, F-ß-hCG and uE3 was 0.91 MoM, 0.94 MoM and 1.05 MoM, respectively. Of the 16 000 pregnant women, there was no statistical difference in the median MoM of triple screening marker at different weeks of gestation (P > 0.05). The positive rate of DS, ES and ONTD in women ≤35 years old (n = 14 972) was 4.03% (603/14 972), 0.36% (54/14 972) and 0.29% (44/14 972) respectively. And in women>35 years old (n = 1 028), the positive rate was 24.51% (252/1 028), 1.95% (20/1 028) and 0.78% (8/1 028), respectively. There was a statistically significant difference of positive rate between the two groups (P < 0.05). (2) 9 cases of DS, 1 case of ES and 1 case of ONTD were found in the high risk group, and 2 cases of DS in the low risk group. The detection rate of DS, ES and ONTD was 9/11, 1/1 and 1/1 respectively; and the positive predictive value was 1.05% (9/855), 1.35% (1/74) and 1.92% (1/52), respectively. (3)The incidence of adverse outcome (group 1) was 1.49 % ( 239/16 000). 7 760 pregnant women in this study were healthy during pregnancy, so were their fetuses (group 2). There were significant differences in the age at delivery, body weight and markers' MoM of STMSTS between the two groups (P < 0.01). (4) In group 1, the rate of abnormal MoM of AFP or F-ß-hCG was 7.95% (19/239) and 23.85% (57/239), and the abnormal rate of MoM of uE3 was 4.18% (10/239). The rate of two abnormal MoM of markers was 5.02% (12/239); the rate that all three MoM were abnormal was 0.84% (2/239). However, in group 2, the rate of two abnormal MoM of markers was 0.14 % ( 11/7 760); and the rate that all three MoM were abnormal was 0. There was a significant difference of abnormal MoM of maternal serum marker between the two groups (P < 0.01). CONCLUSIONS: There is a relationship between abnormal marker of STMSTS and adverse outcomes. STMSTS show a high value in the detection of DS, ES and ONTD.

Nitrogen removal performance and the biocenosis with microalgae consortium Nitrosifying and anammox bacteria in an upflow reactor.

This study introduced an innovative pathway utilizing an algal anaerobic ammonium oxidation (ALGAMMOX) system to treat ammonium wastewater. Lake bottom sludge and anammox sludge were used to cultivate functional microorganisms and microalgae for nitrogen removal in an upflow reactor made of transparent materials. The results showed that the ALGAMMOX system achieved 87.40 % nitrogen removal when the influent NH4 +-N concentration was 100 mg-N/L. Further analysis showed that anammox bacteria Candidatus Brocadia (8.87 %) and nitrosobacteria Nitrosomonas (3.74 %) were crucial contributors, playing essential roles in nitrogen removal. The 16S rRNA gene showed that the anammox bacteria in the sludge transitioned from Candidatus Kuenenia to Candidatus Brocadia. The 18S rRNA gene revealed that Chlamydomonas, Bacillariaceae and Pinnularia were the dominant microalgae in the system at a relative abundance of 7.99 %, 3.64 % and 3.14 %, respectively. This novel approach provides a theoretical foundation for ammonium wastewater treatment.

Investigation of denitrification to Anammox phase transformation performance of Up-Flow anaerobic sludge blanket reactor.

For investigating the microbial community and nitrogen removal performance during the transformation from heterotrophic denitrification (HtDn), mixotrophic denitrification (MtDn), and autotrophic denitrification (AtDn) to anaerobic ammonia oxidation (Anammox), an up-flow anaerobic sludge blanket reactor was constructed by changing the influent substrates and their ratios. The reactor got a total nitrogen removal efficiency (TNRE) of 98.0 % at the molar ratio of carbon, nitrogen, and sulfur sources was 5:8:4 in the MtDn process. In the last phase, the conversion of AtDn to Anammox was successful in 33 days, and a stable TNRE was 87.7 %. The dominant functional bacteria of the microbial communities were Thauera and unclassified_Comamonadaceae in the HtDn process; Thiobacillus, Thauera, Denitratisoma, and Pseudoxanthomonas in the MtDn process; Thiobacillus and Sulfurimonas in the AtDn process; and unclassified_Gemmatimonadaceae, unclassified_SBR1031, and Candidatus_Brocadia in the Anammox process.

The coupling of anammox with microalgae-bacteria symbiosis: Nitrogen removal performance and microbial community.

The partial nitrification-anammox process for ammonia nitrogen wastewater treatment requires mechanical aeration to provide oxygen, which is not conducive to energy saving. The microalgae-bacteria symbiotic system (MaBS) has the advantages of low carbon and energy saving in wastewater biological nitrogen removal. Therefore, this study combined the MaBS with an anammox process to provide oxygen, through the photosynthesis of microalgae instead of mechanical aeration. We investigated the nitrogen removal efficiency and long-term operation of a co-culture system comprising microalgae, nitrifying bacteria (NB), denitrifying bacteria (DnB), and anaerobic ammonium-oxidation bacteria (AnAOB) in a sequencing batch reactor without mechanical aeration. The experiment was divided into three steps: firstly, cultivating NB; then, adding three kinds of microalgae which were Chlorella sp., Anabaena sp., and Navicula sp. to the bioreactor to construct a microalgae-bacteria symbiotic system; finally, adding anammox sludge to construct the anammox and microalgae-bacteria symbiosis (Anammox-MaBS) system. The results demonstrated that nitrification, denitrification, and anammox processes were coupled successfully, and the maximum TN removal efficiency of the stable Anammox-MaBS system was 99.51 % when the concentration of the influent NH4+-N was 100 mg/L. The addition of microalgae in ammonia wastewater promoted the enrichment of DnB and AnAOB, which were Denitratisoma, Haliangium, unclassified_Rhodocyclaceae, and Candidatus_Brocadia. Furthermore, the unique biofilm structure could effectively alleviate the photoinhibition of light-sensitive bacteria, which may be the reason for the long-term adaptation of Candidatus_Brocadia to light conditions. This research can provide a low-cost solution to bacterial photoinhibition in the coexistence system of microalgae and bacteria without mechanical aeration, offering theoretical support for low-carbon and energy-efficient treatment of wastewater.

Complete genome sequence of a highly virulent rabies virus isolated from a rabid pig in south China.

A virulent rabies virus (RABV) strain, GD-SH-01, was isolated from brain tissue of a rabid pig in China. This report describes the first complete genome sequence of a swine-origin RABV strain, and this information will provide important insights into the transmission cycle and genetic diversity of RABV from different hosts in China.

45,X mosaicism in northeast China: a clinical report and review of the literature.

PURPOSE: To explore the prevalence and clinical features, especially the reproductive function, of 45,X mosaicism patients in northeast China. METHODS: GTG-banding was performed on a series of 2,250 patients from our genetic counseling clinic. Each of these patients underwent a physical examination and was interviewed about their medical history and reproductive problems. Literature on 45,X mosaicism was accessed using PubMed and reviewed. RESULTS: The prevalence of 45,X mosaicism in northeast China is 0.36 % (8/2250), and the mosaic karyotype of our study accounted for 61.54 % (8/13) of Turner syndrome cases. This is comparable with studies from Asia, Europe, South America and other regions. The affected patients showed genital abnormalities, abnormal pregnancy or infertility. CONCLUSION: 45,X mosaicism is commonly seen in the genetic counseling clinic. Extensive cytogenetic assessment may improve the detection rate in patients with congenital dysplasia, or history of abnormal pregnancy or infertility. Karyotyping plays a key role in prognosis and assisted reproduction or early surgical treatment.

The performance and microbial communities of Anammox and Sulfide-dependent autotrophic denitrification coupling system based on the gel immobilization.

Anammox and sulfide-dependent autotrophic denitrification (ASDAD) coupling system can improve the nitrogen removal, but high sulfide concentration will affect the activity of anaerobic ammonia-oxidizing bacteria (AnAOB). Gel immobilization technology can enhance the survivability of microorganisms in unsuitable environments. Therefore, in this investigation, gel immobilization technology was applied into the ASDAD coupling system to explore the removal performance and microbial communities. The results showed that the optimal S2-/NO3- was 0.6, under which the best TN removal efficiency was 85.69%. The removal performance of ASDAD coupling system was stable under rapid variations of nitrogen loading rate and sulfide loading rate. Immobilized sludge cubes could attenuate the effects of temperature on AnAOB and sulfide-oxidizing bacteria. Observations of SEM and stereoscope suggested that AnAOB was more likely to exist on the surface of the sludge cubes. Thiobacillus, Candidatus Brocadia, and Candidatus Kuenenia were the main functional bacteria in the coupling system.

The nitrogen removal performance and microbial community on mixotrophic denitrification process.

Sulfur autotrophic denitrifiers and heterotrophic denitrifiers widely exist in aquatic ecosystem, however, the response of sulfide to the microbial community structure in mixotrophic denitrification ecosystem is unknown yet. In this study, the denitrification performance and microbial community were explored by changing the molar ratio of influent C/N/S. From the level of genus, the joint action of Thauera, Pacacoccus, Fusibacter Pseudoxanthomonas, Thiobacillus, Sulfurovum and Sulfurimonas brought about the efficient denitrification performance in the mixotrophic system. Thauera increased from from 0.97% to more than 13%, and the relative abundances of Thiobacillus and Sulfurimonas were about 4.14% and 3.89% separately after adding S2-. The results of this study showed that the denitrification performance could be indeed intensified in the mixotrophic system, among which provided a theoretical basis for establishing an efficient biological nitrogen removal system.

Reproductive outcomes of 3 infertile males with XYY syndrome: Retrospective case series and literature review.

The aim of this study is to evaluate the pregnancy outcomes of males with a 47, XYY karyotype following assisted reproductive treatment.A retrospective study was performed using data from infertile men with 47, XYY at a center for reproductive medicine in 2004 to 2017. Of the 19,842 infertile males treated, a total of 21 showed the 47, XYY karyotype and were included in the present study. Clinical variables were collected. Three men were under treatment with their partner before either in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI).The incidence of 47, XYY in infertile men is 1/945 (21/19842). Most men are azoospermic or severely oligospermic. Three men and their partners underwent IVF or ICSI treatment with fresh ejaculate samples. The fertilization rate was 52.94% to 83.33%. The embryo cleavage rate was 50% to 90%. One man had abnormal sex hormonal levels and his partner had no clinical pregnancy. The other 2 couples had healthy baby boys.Live spermatozoa can be gathered and fertility is possible for infertile males with 47, XYY syndrome when IVF or ICSI treatment is used. It is recommended that genetic counseling is provided in such cases.

Prenatal Diagnosis and Molecular Cytogenetic Characterization of Copy Number Variations on 4p15.2p16.3, Xp22.31, and 12p11.1q11 in a Fetus with Ultrasound Anomalies: A Case Report and Literature Review.

Chromosomal rearrangements, such as duplications/deletions, can lead to a variety of genetic disorders. Herein, we reported a prenatal case with right aortic arch and aberrant left subclavian artery, consisting of a complex chromosomal copy number variations. Routine cytogenetic analysis described the chromosomal karyotype as 46,XY, add (2)(q37) for the fetus. However, the chromosomal microarray analysis (CMA) identified a 22.4 Mb duplication in chromosome 4p16.3p15.2, a 3.96 Mb microduplication in 12p11.1q11, and a 1.68 Mb microdeletion in Xp22.31. Fluorescence in situ hybridization (FISH) using a chromosome 4 painting probe was found to hybridize to the terminal of chromosome 2q on the fetus, thus confirming that the extra genetic materials of chromosome 2 was actually trisomy 4p detected through CMA. Meanwhile, the parental karyotypes were normal, which proved that the add (2) was de novo for fetus. The duplication of Wolf-Hirschhorn syndrome critical region (WHSCR) and X-linked recessive ichthyosis associated with Xp22.31 deletion separately were considered potentially pathogenic causes although other abnormalities involving these syndromes were not observed. For prenatal cases, the combined utilization of ultrasonography, traditional cytogenetic, and molecular diagnosis technology will enhance better diagnostic benefits, offer more detailed genetic counselling, and assess the prognosis of the fetuses.

Prenatal diagnosis and molecular cytogenetic characterization of a small supernumerary marker chromosome (sSMC) inherited from her mosaic sSMC(15) mother and a literature review.

OBJECTIVE: We characterized a maternally inherited small supernumerary marker chromosome (sSMC) derived from chromosome 15 according to prenatal detection and made a review on the prenatal sSMC(15) cases with mosaic maternal inheritance. CASE REPORT: A 29-year-old woman underwent amniocentesis at 19 weeks of gestation due to the high risk of Down syndrome in maternal serum screening. No abnormalities were observed in prenatal ultrasound findings. G-banding analysis revealed a karyotype of 47,XX,+mar. Subsequently, we recalled the couple back for chromosomal analysis. The father's karyotype was normal while the mother's karyotype was 47,XX,+mar[15]/46,XX[35]. Molecular genetic analysis was utilized to identify the marker chromosome. The chromosomal microarray analysis (CMA) results of the mother showed there existed microduplications in the locus of 14q32.33, 15q21.1, 19p12 and Xq26.2, respectively. Then Fluorescence in situ hybridization (FISH) using specific probes for chromosomes 13/21, 14/22, and 15 was applied on the mother and the fetus. And the marker chromosomes for the mother and the fetus were all finally identified as inv dup(15) (D15Z1++, SNRPN-, PML-), which illustrated that the fetus inherited the sSMC(15) from her mother. Finally, a healthy female infant was delivered with no phenotypic abnormalities at 39 weeks. CONCLUSION: The combined utilization of the molecular genetic technologies, such as FISH and CMA, plays a critical role in the identification of the origins and genetic constitutions of sSMC, which would make a significant contribution to genetic counseling and prenatal diagnosis.

Deletion of b1/b3 shows risk for expanse of Yq microdeletion in male offspring: Case report of novel Y chromosome variations.

RATIONALE: This study aimed to report 1 family case with novel Y chromosome structural variations by an established next-generation sequencing (NGS) method using unique STSs. PATIENT CONCERNS: The case studied was from a family with a father and son (the proband). G-band staining was used for karyotype analysis. Y chromosome microdeletions were detected by sequence-tagged site (STS)-PCR analysis and a new NGS screening strategy. DIAGNOSES: Semen analysis showed that the proband was azoospermic. The patient had an abnormal karyotype (45,X[48%]/46,XY[52%]). His father exhibited a normal karyotype. STS-PCR analysis showed that the proband had a deletion of the AZFb+c region, and his father had no deletion of STS markers examined. The sequencing method revealed that the patient had DNA sequence deletions from nt 20099846 to nt 28365090 (8.3 Mb), including the region from yel4 to the Yq terminal, and his father exhibited a deletion of b1/b3 and duplication of gr/gr. INTERVENTIONS: The proband was advised to undergo genetic counseling, and consider the use of sperm from a sperm bank or adoption to become a father. OUTCOMES: The proband was azoospermic. AZFc partial deletions may produce a potential risk for large AZFb+c deletions or abnormal karyotypes causing spermatogenic failure in men. LESSONS: The NGS method can be considered a clinical diagnostic tool to detect Y chromosome microdeletions. The partial AZFc deletions and/or duplications can be a risk of extensive deletions in offspring.

Cytogenetic and molecular detection of a rare unbalanced Y;3 translocation in an infertile male: A case report.

INTRODUCTION: The infertile male individuals carrying the Y-autosome translocations are seldom reported in clinic. Herein, we described a severe oligozoospermic male with rare unbalanced Y;3 translocation transmitted through 3 generations. PATIENT CONCERNS: A 33-year-old Chinese male was referred for infertility consultation in our center after 10 years' primary infertility. He was diagnosed as severe oligozoospermia according to the semen analysis. DIAGNOSIS: G-banding analysis initially described the karyotype as 46, XY, add (3) (p26) for the patient, and his wife's karyotype was 46, XX. The chromosomal microarray analysis identified 3.81Mb and 0.29Mb duplications in Yq11.223q11.23 and Yq12, separately. No deletions were detected in azoospermia factors (AZF)a, AZFb and AZFc. Fluorescence in situ hybridization analysis further confirmed the existence of sex-determining region Y gene and verified that Yq12 was translocated to the terminal short arm of chromosome 3(3p26). INTERVENTIONS: The couple chose intracytoplasmic sperm injection to get their offspring. The wife underwent amniocentesis for cytogenetic analysis but suffered termination of pregnancy due to premature rupture of membranes. OUTCOMES: The karyotype of the patient was finally described as 46, X, der(3)t(Y;3)(q11.22;p26). His father and the aborted fetus showed the same karyotypes as the patient. CONCLUSION: Our study not only enriched the karyotype-phenotype correlation of Y-autosome translocation, but also strengthened the critical roles of molecular genetic techniques in identifying the chromosomal breakpoints and regions involved.

Molecular cytogenetic characterization of a mosaic small supernumerary marker chromosome derived from chromosome Y in an azoospermic male: A case report.

RATIONALE: Small supernumerary marker chromosomes (sSMCs) can be usually discovered in the patients with mental retardation, infertile couples, and prenatal fetus. We aim to characterize the sSMC and explore the correlation between with sSMC and male infertility. PATIENT CONCERNS: A 26-year-old Chinese male was referred for infertility consultation in our center after 1 year of regular unprotected coitus and no pregnancy. DIAGNOSIS: Cytogenetic G-banding analysis initially described a mosaic karyotype 47,X,Yqh-,+mar[28]/46,X,Yqh-[22] for the proband, while his father showed a normal karyotype. The chromosome microarray (CMA) analysis showed there existed a duplication of Yp11.32q11.221, a deletion of Yq11.222q12, a duplication of 20p11.1 for the patient. Azoospermia factor (AZF) microdeletion analysis for the patient showed that he presented a de novo AZFb+c deletion. Fluorescence in situ hybridization further confirmed the sSMC was an sSMC(Y) with SRY signal, Y centromere, and Yq deletion. INTERVENTIONS: The patient would choose artificial reproductive technology to get his offspring according to the genetic counseling. OUTCOMES: The sSMC in our patient was proved to be an sSMC(Y), derived from Yq deletion. The spermatogenesis failure of the proband might be due to the synthetic action of sSMC(Y) mosaicism and AZFb+c microdeletion. LESSONS: It is nearly impossible to detect the chromosomal origin of sSMC through traditional banding techniques. The molecular cytogenetic characterization could be performed for identification of sSMC so that comprehensive genetic counseling would be offered.