Reproductive outcomes of intracytoplasmic sperm injection using testicular sperm and ejaculated sperm in patients with AZFc microdeletions: a systematic review and meta-analysis.

Studies have explored the assisted reproductive technology (ART) outcomes of Y-chromosome azoospermia factor c (AZFc) microdeletions, but the effect of sperm source on intracytoplasmic sperm injection (ICSI) remains unknown. To determine the ART results of ICSI using testicular sperm and ejaculated sperm from males with AZFc microdeletions, we searched Embase, Web of Science, and PubMed to conduct a systematic review and meta-analysis. The first meta-analysis results for 106 cycles in five studies showed no significant differences in the live birth rate between the testicular sperm group and the ejaculated sperm group (risk ratio: 0.97, 95% confidence interval [CI]: 0.73-1.28, P = 0.82). The second meta-analysis of 106 cycles in five studies showed no difference in the abortion rate between the testicular sperm group and ejaculated sperm group (risk ratio: 1.06, 95% CI: 0.54-2.06, P = 0.87). The third meta-analysis of 386 cycles in seven studies showed no significant difference in clinical pregnancy rates between the testicular sperm group and the ejaculated sperm group (risk ratio: 1.24, 95% CI: 0.66-2.34, P = 0.50). Inevitable heterogeneity weakened our results. However, our results indicated that testicular sperm and ejaculated sperm yield similar ART outcomes, representing a meaningful result for clinical treatment. More properly designed studies are needed to further confirm our conclusions.

Risk and mechanism of glucose metabolism disorder in the offspring conceived by female fertility maintenance technology.

Although female fertility maintenance technology (FFMT) provides an effective option for preserving fertility in patients with cancer suffering from fertility loss due to cancer treatment, previous studies have shown that the technique has certain potential risks and requires an assessment of the health status of the offspring since FFMT may lead to glucose metabolism disorder in offspring mice. The present animal study examined the glucose metabolism of adult mice offspring born from ovarian tissue cryopreservation and orthotopic allotransplantation. The mice were divided into three groups: normal, fresh ovary transplantation, and cryopreserved ovary transplantation. We recorded fasting blood glucose, glucose tolerance, and fasting serum insulin level for six months. Liver DNA, RNA, and proteins were extracted to detect the interaction between DNA methylation and Grb10 expression and insulin signaling pathway factors such as P-IGF1R, P-IRS2, P-AKT, and Grb10. Female recipient mice that received FFMT could successfully give birth after mating. The average litter size and total litter size of the cryopreserved and fresh groups showed marked differences compared with the normal group. Compared with the normal group, the fasting blood glucose and fasting serum insulin levels were higher in the cryopreserved and fresh groups. The mRNA and protein expressions of Grb10 were higher in the fresh and cryopreserved groups. Compared with the normal group, the DNA methylation status of four of the 11 sites of the Grb10 promoter was lower in the cryopreserved group. Grb10 overexpression inhibited the downstream phosphorylation protein factor expression (p-IGF-1R, p-IRS2, and p-Akt) of the IGF-1R signaling pathway. Female fertility maintenance technology (FFMT), including ovarian tissue cryopreservation (OTC), and orthotopic allotransplantation techniques might lead to glucose metabolism disorders in offspring mice.

Histone H3 methylation orchestrates transcriptional program in mouse spermatogenic cell line.

Changes in histone modifications always correlate with altered transcriptional activities of genes. Recent studies have shown that the mutation of certain lysine residues to methionine in the histone variant H3.3 can act as a valuable tool to reduce specific H3 methylation levels. In our study, we used the mouse spermatogenic cell line GC-2 as a model to generate cells stably expressing H3.3 K4, H3.3 K9, H3.3 K27, and H3.3 K36M. The expression of these H3.3 K-to-M mutants influenced the expression of different subsets of genes, and a total of 891 differentially expressed genes were identified through global gene expression profiling. Moreover, the H3.3 K-to-M transgenes, especially H3.3 K36M, impacted the expression of endogenous retrovirus ERVK. This study gives a global view of how different H3 modifications regulate transcriptomes in spermatogenic cell lines, and identifies potential targets of H3 modifications in male germ line.

Seminal plasma neutral alpha-glucosidase activity as an early predictor of patency and natural pregnancy after microsurgical vasoepididymostomy.

Microsurgical vasoepididymostomy (MVE) is recommended as a first-line option for treatment of epididymal obstructive azoospermia (EOA). However, early indicators for predicting patency and natural pregnancy are unclear. Our aim was to explore the early predictive value of seminal plasma neutral alpha-glucosidase (NAG) activity for patency and natural pregnancy after MVE. Eighty-four patients with EOA who underwent MVE were enrolled in this study. The post-operative patency and natural pregnancy rates were 60.71% and 33.33% respectively. The presence of motile epididymal spermatozoa at the anastomosis site and NAG activity measured at the first month after MVE were early and independent predictors of patency and natural pregnancy. The areas under the receiver operating characteristic curves (AUCs) of NAG activity for prediction of patency and natural pregnancy were 0.78 (95% confidence interval [CI]: 0.68-0.88) and 0.82 (95% CI: 0.73-0.92). The best cut-off values of NAG activity for predicting patency and pregnancy were 15.9 and 17.0 m IU/ejaculate respectively. In conclusion， NAG activity measured at the first month after MVE is an early and independent predictor of patency and natural pregnancy.

[Characteristics of sperm donors and the eligibility rate of screening for sperm donation].

OBJECTIVE: To investigate the relationship of the characteristics of sperm donors with the results of screening and provide some reference for the screening of sperm donors. METHODS: We statistically analyzed the screening data about 12 362 sperm donors at the Guangdong Human Sperm Bank from January 2003 to June 2017 and the relationship of the eligibility rate of screening with the donors' age, education, occupation, marriage, and fatherhood. RESULTS: Of the 12 362 sperm donors, 3 968 (32.1%) met the standards of semen quality and 3 127 (25.3%) filled all the requirements of sperm donation. The eligibility rate of screening was 27.7% in the donors aged 20－24 years, 24.3% in those aged 25－29 years, 23.8% in those aged 30－34 years, and 17.5% in those aged =≥35 years (P < 0.01); 23.5% in the senior high school students, 24% in the junior college students, 25.9% in the undergraduates, and 30.3% in the postgraduates (P < 0.01); 29.3% in the students versus 22.9% in the others (P < 0.01), 41.5% in the married versus 20.7% in the unmarried (P < 0.01), and 45.6% in the fathers versus 20.9% in the childless husbands (P < 0.01). CONCLUSIONS: A higher eligibility rate of screening was found among the sperm donors aged <35 years or with a bachelor's or higher degree, particularly among students.

Cellular Mechanism Underlying Hydrogen Sulfide Mediated Epithelial K+ Secretion in Rat Epididymis.

As a novel gasotransmitter, hydrogen sulfide (H2S) elicits various physiological actions including smooth muscle relaxation and promotion of transepithelial ion transport. However, the pro-secretory function of H2S in the male reproductive system remains largely unclear. The aim of this study is to elucidate the possible roles of H2S in modulating rat epididymal intraluminal ionic microenvironment essential for sperm storage. The results revealed that endogenous H2S-generating enzymes cystathionine ß-synthetase (CBS) and cystathionine γ-lyase (CSE) were both expressed in rat epididymis. CBS located predominantly in epithelial cells whilst CSE expressed primarily in smooth muscle cells. The relative expression level of CBS and CSE escalated from caput to cauda regions of epididymis, which was paralleled to the progressively increasing production of endogenous H2S. The effect of H2S on epididymal epithelial ion transportation was investigated using short-circuit current (I SC), measurement of intracellular ion concentration and in vivo rat epididymal microperfusion. Our data showed that H2S induced transepithelial K+ secretion via adenosine triphosphate-sensitive K+ (KATP) channel and large conductance Ca2+-activated K+ (BKCa) channel. Transient receptor potential vanilloid 4 (TRPV4) channel-mediated Ca2+ influx was implicated in the activation of BKCa channel. In vivo studies further demonstrated that H2S promoted K+ secretion in rat epididymal epithelium. Inhibition of endogenous H2S synthesis caused a significant decrease in K+ concentration of cauda epididymal intraluminal fluid. Moreover, our data demonstrated that high extracellular K+ concentration actively depressed the motility of cauda epididymal sperm in a pH-independent manner. Collectively, the present study demonstrated that H2S was vital to the formation of high K+ concentration in epididymal intraluminal fluid by promoting the transepithelial K+ secretion, which might contribute to the maintenance of the cauda epididymal sperm in quiescent dormant state before ejaculation.

Association of decreased spermatozoa omega-3 fatty acid levels and increased oxidative DNA damage with varicocele in infertile men: a case control study.

Varicocele is commonly associated with male infertility because it impairs normal sperm morphology and activity. Polyunsaturated fatty acids (PUFA) are important determinants of sperm cell structure and function, but their relationship with varicocele remains unclear. The aim of the present study was to investigate the PUFA composition in spermatozoa of infertile men with varicocele and to evaluate the potential relationship between PUFA and varicocele. This case control study recruited 92 infertile men with varicocele, 99 infertile men without varicocele and 95 fertile male control subjects. Semen morphology and activity parameters were assessed and seminal plasma 8-hydroxy-2-deoxyguanosine (8-OHdG) content was determined by ELISA. Sperm concentrations of omega-3 and omega-6 fatty acids were measured by gas chromatography. Infertile men with varicocele had lower concentrations of omega-3 PUFA, higher omega-6:omega-3 PUFA ratios and greater oxidative DNA damage in spermatozoa compared with infertile men without varicocele and normal subjects. The degree of varicocele and DNA damage was associated with decreased omega-3 PUFA concentrations and semen quality in infertile men with varicocele. The findings suggest that omega-3 PUFA deficiency could be implicated in varicocele-associated infertility, and highlight the need for intervention trials to test the usefulness of omega-3 supplementation in reducing sperm abnormalities in infertile men with varicocele.

The reference values for semen parameters of 1213 fertile men in Guangdong Province in China.

Semen samples were collected from 1213 fertile men whose partners had a time-to-pregnancy (TTP) ≤12 months in Guangdong Province in Southern China, and semen parameters including semen volume, sperm concentration, total counts, motility, and morphology were evaluated according to the World Health Organization (WHO) 2010 guideline. All semen parameters analyzed were normal in ~62.2% of the total samples, whereas ~37.8% showed at least one of the semen parameters below normal threshold values. The fifth centiles (with 95% confidence intervals) were 1.3 (1.2-1.5) ml for semen volume, 20 × 10 6 (18×10 6 -20×10 6 ) ml-1 for sperm concentration, 40 × 10 6 (38×10 6 -44×10 6 ) per ejaculate for total sperm counts, 48% (47%-53%) for vitality, 39% (36%-43%) for total motility, 25% (23%-27%) for sperm progressive motility, 5.0% (4%-5%) for normal morphology. The pH values ranged from 7.2 to 8.0 with the mean ± standard deviation at 7.32 ± 0.17. No effects of age and body mass index were found on semen parameters. Occupation, smoking and alcohol abuse, varicocele appeared to decrease semen quality. Sperm concentration, but not sperm morphology, is positively correlated with TTP, whereas vitality is negatively correlated with TTP. Our study provides the latest reference values for the semen parameters of Chinese fertile men in Guangdong Province, which are close to those described in the new WHO guidelines (5 th Edition).

[High-frequency ultrasonography for epididymal stasis after vasectomy].

OBJECTIVE: To explore the clinical application of high-frequency ultrasound in the diagnosis and treatment of epididymal stasis after vasectomy. METHODS: We retrospectively studied the sonographic characteristics of 23 cases of epididymal stasis treated by vasectomy, which were divided into a mild (n = 5), a moderate (n = 11) and a severe group (n = 7) according to the results of color Doppler flow imaging. We analyzed the significance of high-frequency ultrasonography in the treatment of epididymal stasis after vasectomy. RESULTS: High-frequency ultrasonography revealed 14 cases of increased bilateral epididymal volume, 6 cases of left epididymal thickening and 3 cases of right epididymal thickening, mainly the thickening of the epididymal body and tail. After conservative treatment, 18 of the epididymal stasis cases (5 mild, 11 moderate and 2 severe) were improved, and the other 5 severe cases significantly relieved and discharged from hospital following conservative treatment combined with vasectomy reversal. CONCLUSION: Post-vasectomy epididymal stasis has typical sonographic characteristics, and high-frequency ultrasonography has an extremely important application value in the clinical classification, diagnosis and treatment of the disease.

[Semen quality of normal fertile men].

OBJECTIVE: To study the semen quality of confirmed fertile men with the time to pregnancy (TTP) of 12 months or less, and analyze the correlation of semen parameters with TTP. METHODS: We recruited 1 152 men aged 18 -50 years and with TTP of 12 months. We used questionnaire investigation, physical examination and semen analysis, calculated the normal reference ranges of semen parameters, and analyzed the effects of TTP and other biological characteristics on semen quality. RESULTS: Seminal examinations showed that the semen volume was 1.5 ml, sperm concentration 17 million per ml, total sperm number 39 million per ejaculate, total sperm motility 41%, progressive sperm motility 33%, normal sperm morphology 5.0%, sperm vitality 53%, and total number of progressively motile sperm 17 million per ejaculate in the 1 152 subjects. In comparison, the total number of progressively motile sperm was significantly smaller in those aged > or = 35 years (P < 0.01), morphologically normal sperm were markedly less in those with BMI > 25.0 (P < 0.01), and sperm concentration, total number of sperm, morphologically normal sperm rate and total number of progressively motile sperm were remarkably lower in those with BMI < 18. 5 (P < 0.05). Cigarette smoking significantly reduced the sperm volume (P < 0.01), and drinking (> 10 g per week) decreased the sperm volume and the total numbers of sperm, morphologically normal sperm and progressively motile perm (P < 0.05). TTP was negatively correlated with sperm concentration, total number of sperm and total number of progressively motile sperm (P < 0.01). The subjects with sexual abstinence of 5 -7 days showed markedly higher sperm concentration, total number of sperm and total number of progressively motile sperm than others (P < 0.01). CONCLUSION: In normal fertile men, the fifth centile of semen parameters is close to the WHO normal reference range, TTP is negatively correlated with semen quality, and age, BMI and lifestyle may affect some of the semen parameters.

[Chromosome abnormalities and Y chromosome microdeletions in patients with the azoospermia and cryptozoospermia].

OBJECTIVE: To study the incidence of the chromosome abnormalities and Y chromosome microdeletions in Chinese patients with azoospermia and cryptozoospermia. METHODS: Conventional chromosomal karyotyping was used to analyze the chromosome abnormalities. Genomic DNA was extracted from peripheral blood samples and multiplex polymerase chain reactions (PCR) analyses were performed using specific primers to confirm the presence or absence of Y chromosome microdeletions. A total of 997 patients with azoospermia and cryptozoospermia were enrolled in the study. RESULTS: The incidence of chromosome abnormalities in the patient with azoospermia and cryptozoospermia was 28.4%. The major abnormal karyotypes included 47,XXY, 46,XY (Y>G), 46,XX, chimera and translocations. The incidence of the Y chromosome microdeletions was 17.4%. They were mainly found in the karyotypes of 46,XY and 46,XY (Y>G). CONCLUSION: Chromosome abnormalities were the most common hereditary causes of the patients with azoospermia and cryptozoospermia. The incidence of Y chromosome microdeletion was higher in the patients with karyotype of 46,XY and 46,XY (Y>G). Therefore, detection of the AZF microdeletion in these patients is helpful to determine the etiology and avoid the unnecessary treatment and vertical transmission of the genetic defects.

[Relationship between testis volume and types of spermatogenic cells from testicular biopsy in patients with azoospermia or cryptozoospermia].

OBJECTIVE: To study the relationship between the testis volume and types of spermatogenic cells derived from testicular biopsy in patients with azoospermia or cryptozoospermia. METHODS: We collected testicular pathological biopsies from 492 infertile patients with azoospermia or cryptozoospermia reported in our hospital, classified them according to the testicular histological classification methods in WHO Manual for Standardized Investigation, Diagnosis and Management of the Infertile Male, and analyzed the relationship of the testis volume with the results of semen analyses and testicular histology. RESULTS: Of the 492 cases, 90.5% (445/492) were azoospermia and 9.5% (47/492) cryptozoospermia; mature spermatozoa were present in the seminiferous tubules in 17.9% (88/492) but absent in 42.9% (211/492), and Sertoli cell-only syndrome indicated in 39.2% (193/492); the testis volume was < or = 10 ml in 38.6% (190/492) and < or = 5 ml in 7.9% (39/492). Cryptozoospermia was detected in 14.8% (13/88) of those with mature spermatozoa in the seminiferous tubules, in 11.4% (24/211) of those without, and in 5.2% (10/193) of those with Sertoli cell-only syndrome, with a significantly lower rate in the latter group than in the former two (P < 0.05). CONCLUSION: Spermatogenesis of the testis may be focal and difficult to be completely reflected by a single testicular biopsy, and it may exist even if the testis volume is significantly below the reference value. The indications for testicular biopsy should not be improperly expanded. The WHO testicular histological classification methods have provided a convenient and effective guidance for further clinical examinations and establishment of a protocol.