[Chromosome polymorphisms and their influence on semen quality and sperm DNA integrity in males undergoing IVF/ICSI].

OBJECTIVE: To investigate the incidence of chromosome polymorphisms and their influence on semen quality and sperm DNA integrity in male patients receiving in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). METHODS: We retrospectively analyzed the chromosomal karyotypes and the types and incidence rate of chromosome polymorphisms in 2 370 male patients undergoing IVF/ICSI between June 2016 and June 2018. We classified the patients into groups A (with variation in the secondary constriction region in the autosomal long arm), B (with variation in the short arm of the D/G group chromosomes), C (with interbrachial inversion of chromosome 9) and D (with Y chromosome polymorphisms), and compared the semen parameters and sperm DNA fragmentation indexes (DFI) between the patients with chromosome polymorphisms and those with normal chromosomes. RESULTS: Totally, 154 (6.50%) of the patients undergoing IVF/ICSI were found with chromosome polymorphisms, including 34 cases of secondary constriction variation in the long arm of the autosome (1.43% ï¼»34/2 370ï¼½, 22.08% ï¼»34/154ï¼½), 82 cases of short arm polymorphisms of the D/G group chromosomes (3.46% ï¼»82/2 370ï¼½, 53.25% ï¼»82/154ï¼½), 26 cases of interbrachial inversion of chromosome 9 (1.10% ï¼»26/2 370ï¼½, 16.88% ï¼»26/154ï¼½), 10 cases of Y chromosome polymorphisms (0.42% ï¼»10/2 370ï¼½, 6.50% ï¼»10/154ï¼½), and 2 cases of mixed chromosome polymorphisms (0.08% ï¼»2/2 370ï¼½, 1.42% ï¼»2/154ï¼½). The total sperm count was lower in group D than in the other polymorphism groups and the normal chromosome group, but with no statistically significant difference among the five groups (P > 0.05). The sperm progressive motility was also lower in group D than in the other five groups, with statistically significant difference from group B (27.5 ± 13.5 vs. 41.5 ± 21.1, P = 0.027), but not from the other groups (P > 0.05). No statistically significant difference was observed in the sperm DFI between the polymorphism groups and the normal chromosome group (P > 0.05), or among the polymorphism groups (P > 0.05). The proportion of normal semen was lower in group D than in the other four groups, but with no statistically significant difference among the five groups (P > 0.05). The incidence rate of asthenospermia was higher in group D than in the other four groups, but with no statistically significant difference among the five groups (P > 0.05), and so was that of oligoasthenospermia, with statistically significant difference from the normal chromosome group (30.0% vs 8.0%, P = 0.041), but not from the other polymorphism groups (P > 0.05). CONCLUSIONS: Short arm polymorphisms of the D/G group chromosomes are the most common type of chromosome polymorphisms in male patients undergoing IVF/ICSI. Polymorphisms of the Y chromosome have a negative effect on semen quality, while those of the other chromosomes do not significantly affect semen quality and sperm DNA integrity.

AZFa Microdeletions: Occurrence in Chinese Infertile Men and Novel Deletions Revealed by Semiconductor Sequencing.

OBJECTIVE: To evaluate the frequency of azoospermia factor (AZFa) microdeletions among infertile men and establish a new high-throughput sequencing method to detect novel deletion types. MATERIALS AND METHODS: A total of 3731 infertile men were included. Karyotype analysis was performed using G-band staining of peripheral blood lymphocytes. Polymerase chain reaction (PCR) amplification using specific sequence-tagged sites (STS) was performed to screen for AZF region microdeletions of the Y chromosome. A novel semiconductor sequencing method was established to detect high-resolution AZFa microdeletions. RESULTS: Of 3731 infertile men, 341 (9.14%) had microdeletions in AZFa, AZFb, or AZFc. Thirteen of these (3.81%) had a deletion in the AZFa region (mean age: 27.3 ± 4 years, range: 22-34), which included 12 subjects with a normal karyotype (46, XY) and 1 with Klinefelter syndrome (47, XXY). Four of 10 subjects with complete AZFa microdeletions (sY86 and sY84 loss) underwent semiconductor sequencing. They all had DNA sequence deletions from nt 14469266 to 15195932, whereas their fathers had no deletions. One subject with partial AZFa microdeletion (sY86 loss) and his father underwent semiconductor sequencing and STS-PCR analysis. The same deletion (sY86 loss with DNA sequence deletion from nt 14469266 to 14607672) was identified in both subjects. Forty sperm donators and 50 infertile men showed no AZFa microdeletions by either method. CONCLUSION: AZFa deletions are present at a low frequency in men with azoospermia or oligozoospermia. Novel sequencing methods can be used for these patients to reveal high-resolution AZFa microdeletions.

Complete Azoospermia Factor b Deletion of Y Chromosome in an Infertile Male With Severe Oligoasthenozoospermia: Case Report and Literature Review.

OBJECTIVE: To report on a male patient with complete deletion of azoospermia factor b (AZFb) who presented with severe oligoasthenozoospermia, but who successfully fathered a child via intracytoplasmic sperm injection (ICSI). MATERIALS AND METHODS: Karyotype analysis of peripheral blood lymphocytes was performed by standard G-banding. Y chromosome microdeletions were detected by multiplex polymerase chain reaction amplification using AZF-specific, sequence-tagged site markers. The ICSI procedure was performed using ejaculated motile spermatozoa. RESULTS: Cytogenetic analysis of the patient revealed a normal male karyotype, 46,XY. Multiplex polymerase chain reaction screening showed complete deletion of AZFb demonstrated by the absence of specific sequence-tagged site markers sY121, sY127, sY134, and sY143. Following successful ICSI, an ultrasound scan of the patient's partner revealed a single pregnancy with cardiac activity. A healthy boy was born by cesarean section at 38 weeks of gestation. Genetic testing 2 years later revealed that the infant had inherited his father's AZFb deletion. CONCLUSION: Evidence from this case supports the fact that carriers of AZFb deletions can sometimes produce spermatozoa and father a son with the same AZFb deletion. This possibility reinforces the need for genetic counseling in patients with Y chromosome microdeletions.

Effect of sperm DNA fragmentation on clinical outcomes for Chinese couples undergoing in vitro fertilization or intracytoplasmic sperm injection.

Objective To investigate the effect of sperm DNA fragmentation on the fertilization rate, embryo development and pregnancy outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in a cohort of Chinese couples. Methods Infertile couples that had undergone assisted reproductive technology at our centre between January 2011 and December 2013 were included in this retrospective study. Fractions of prepared sperm samples were evaluated for sperm DNA fragmentation on the day of oocyte recovery. Results Of the 550 couples selected, 415 had undergone IVF and 135 ICSI. Sperm DNA fragmentation rate was significantly negatively correlated with the fertilization rate in the ICSI cycles but not the IVF cycles. No association was found between sperm DNA fragmentation and cleavage rate or good quality embryo formation rates in IVF or ICSI cycles. Receiver operating characteristic (ROC) curve analysis showed that the sperm DNA fragmentation rate was a statistically significant prognostic indicator of the clinical fertilization rate in ICSI cycles; a rate > 22.3% was associated with a lower fertilization rate following ICSI compared with a rate ≤ 22.3%. Conclusions High values of sperm DNA fragmentation were associated with a low fertilization rate following ICSI but were not associated with alterations in pregnancy or live birth rates in either ICSI or IVF in this cohort of Chinese couples.