Identification of risk genes in Chinese nonobstructive azoospermia patients based on whole-exome sequencing.

Nonobstructive azoospermia (NOA) is a severe condition in infertile men, and increasing numbers of causative genes have been identified during the last few decades. Although certain causative genes can explain the presence of NOA in some patients, a proportion of NOA patients remain to be addressed. This study aimed to investigate potential high-risk genes associated with spermatogenesis in idiopathic NOA patients by whole-exome sequencing. Whole-exome sequencing was performed in 46 male patients diagnosed with NOA. First, screening was performed for 119 genes known to be related to male infertility. Next, further screening was performed to determine potential high-risk causative genes for NOA by comparisons with 68 healthy male controls. Finally, risk genes with high/specific expression in the testes were selected and their expression fluctuations during spermatogenesis were graphed. The frequency of cystic fibrosis transmembrane conductance regulator (CFTR) gene pathogenic variant carriers was higher in the NOA patients compared with the healthy controls. Potential risk genes that may be causes of NOA were identified, including seven genes that were highly/specifically expressed in the testes. Four risk genes previously reported to be involved in spermatogenesis (MutS homolog 5 [MSH5], cilia- and flagella-associated protein 54 [CFAP54], MAP7 domain containing 3 [MAP7D3], and coiled-coil domain containing 33 [CCDC33]) and three novel risk genes (coiled-coil domain containing 168 [CCDC168], chromosome 16 open reading frame 96 [C16orf96], and serine protease 48 [PRSS48]) were identified to be highly or specifically expressed in the testes and significantly different in the 46 NOA patients compared with 68 healthy controls. This study on clinical NOA patients provides further evidence for the four previously reported risk genes. The present findings pave the way for further functional investigations and provide candidate risk genes for genetic diagnosis of NOA.

Poor morphology of inner cell mass increases birth weight and large for gestational age.

RESEARCH QUESTION: Can inner cell mass (ICM) and trophectoderm morphological grading, especially ICM and trophectoderm graded C, affect perinatal outcomes? DESIGN: A retrospective review of medical records of 8946 singletons delivered from vitrified-warmed single blastocyst transfer cycles between January 2009 and December 2020. RESULTS: Inner cell mass graded C had a higher adjusted birth weight than ICM graded A (0.61 ± 1.06 versus 0.48 ± 1.06; P = 0.025). Large for gestational age (LGA) increased with decreasing ICM morphological grading (18.96%, 21.88% and 23.38%; grade B versus grade A, P = 0.013; grade C versus grade A, P = 0.036) (P < 0.025 was considered statistically significant for multiple pairwise comparisons). Linear regression analysis suggested that ICM morphological grading was significantly associated with adjusted birth weight, with grade C increasing adjusted birth weight compared with grade A (ß 0.13, 95% CI 0.00 to 0.25, P = 0.043) (P < 0.05 was considered statistically significant for linear regression). Logistic regression analysis suggested that ICM morphological grading was significantly associated with LGA, with grade C increasing LGA compared with grade A (adjusted OR 1.37, 95% CI 1.03 to 1.81). Moreover, blastocysts with ICM graded C had a higher chance of being a male infant compared with ICM graded A (adjusted OR 1.32, 95% CI 1.04 to 1.68). CONCLUSIONS: Inner cell mass morphological grading was significantly associated with adjusted birth weight and LGA. Poor ICM graded C increased birth weight and LGA.

FAM9B serves as a novel meiosis-related protein localized in meiotic chromosome cores and is associated with human gametogenesis.

Meiosis is a complex process involving the expression and interaction of numerous genes in a series of highly orchestrated molecular events. Fam9b localized in Xp22.3 has been found to be expressed in testes. However, FAM9B expression, localization, and its role in meiosis have not been previously reported. In this study, FAM9B expression was evaluated in the human testes and ovaries by RT-PCR, qPCR, and western blotting. FAM9B was found in the nuclei of primary spermatocytes in testes and specifically localized in the synaptonemal complex (SC) region of spermatocytes. FAM9B was also evident in the follicle cell nuclei and diffusely dispersed in the granular cell cytoplasm. FAM9B was partly co-localized with SYCP3, which is essential for both formation and maintenance of lateral SC elements. In addition, FAM9B had a similar distribution pattern and co-localization as γH2AX, which is a novel biomarker for DNA double-strand breaks during meiosis. All results indicate that FAM9B is a novel meiosis-associated protein that is co-localized with SYCP3 and γH2AX and may play an important role in SC formation and DNA recombination during meiosis. These findings offer a new perspective for understanding the molecular mechanisms involved in meiosis of human gametogenesis.

Obstetric and neonatal outcomes after the transfer of vitrified-warmed blastocysts developing from nonpronuclear and monopronuclear zygotes: a retrospective cohort study.

OBJECTIVE: To evaluate the obstetric and neonatal outcomes after the transfer of vitrified-warmed single blastocysts developing from nonpronuclear (0PN) and monopronuclear (1PN) zygotes. DESIGN: Cohort study. SETTING: Affiliated hospital. PATIENT(S): This study was a retrospective analysis of 435 0PN and 281 1PN vitrified-warmed single blastocyst transfers, and 151 0PN and 75 1PN singletons, compared with 13,167 two-pronuclear (2PN) vitrified-warmed single blastocyst transfers and 4,559 2PN singletons, respectively. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Pregnancy rate (PR), abortion rate (AR), live birth rate (LBR), and singleton birthweight were the primary outcome measures. RESULT(S): PR, AR, and LBR were similar when compared between the 0PN and 2PN groups after vitrified-warmed blastocyst transfer. However, the 0PN group had a higher birthweights, higher z scores, and a greater proportion of very large for gestational age newborns. When comparing the 1PN and 2PN groups, we found that the PR was similar whereas the AR was higher and the LBR was lower. No differences were detected in the other neonatal outcomes. CONCLUSION(S): The results of the present study show that the transfer of 2PN blastocysts should be prioritized because of a higher AR and a lower LBR after 1PN blastocyst transfers and a higher birthweight after 0PN blastocyst transfers when compared with 2PN blastocyst transfers. Our data indicate the need for concern about the safety of 1PN and 0PN embryo transfers.

Ran-binding protein 3 is associated with human spermatogenesis and male infertility.

Ran-binding protein 3 (RanBP3) is a Ran-interacting protein, which participates in the Ran GTPase system in cancer cell biology. However, the expression pattern and physiological role of RanBP3 remain largely unknown. In this study, we found that RanBP3 was expressed in human testes and localised to spermatogonium and spermatocyte of germ cells. In subcellular structure, its localisation is in the nucleus and cytoplasm. Interestingly, compared with normal groups, RanBP3 expression was lower in groups of patients with Maturation Arrest (MA) and Sertoli cell-only syndrome (SCO) when considered by the Johnson Score. RanBP3 expression in the MA group and SCO groups was dramatically lower than that in the normal control group. Studies have shown that RanBP3, which is one of the helper factors of Ran, is mainly participate in the nucleocytoplasmic transport of cells. RanBP3 helps Ran to achieve some functions such as nucleocytoplasmic transport, spindle assembly during mitosis and nuclear assembly after mitosis. Consequent changes in the expression of RanBP3 may associate with human spermatogenesis disorders and male infertility. The identification and characterisation of RanBP3 enhances our understanding of the molecular mechanisms underpinning its function in human spermatogenesis and male infertility.

Artificial oocyte activation improves cycles with prospects of ICSI fertilization failure: a sibling oocyte control study.

RESEARCH QUESTION: Does artificial oocyte activation improve clinical outcomes for patients at risk of intracytoplasmic sperm injection (ICSI) fertilization failure? DESIGN: In this study, sibling oocytes from patients with previous ICSI failure or severe teratozoospermia were divided equally into two groups, half for artificial oocyte activation (AOA) with ionomycin after conventional ICSI and the other half for conventional ICSI only (non-AOA). The fertilization rates, cleavage rates, transferable embryo rates and blastulation rates of the two groups were compared first; the clinical pregnancy and live birth rates were also compared to assess the efficiency and safety of AOA. RESULT: The outcomes of the AOA group were significantly better than those of the conventional ICSI group in terms of the fertilization (50.38% versus 33.86%, respectively, P < 0.001), cleavage (59.16% versus 39.04%, respectively, P < 0.001) and transferable embryo rates (43.51% versus 26.69%, respectively, P < 0.001). The blastulation (43.53% versus 36.11%, respectively), implantation (26.83% versus 15.79%, respectively), clinical pregnancy (38.46% versus 25%, respectively) and live birth rates (38.46% versus 16.67%, respectively) were not significantly different. CONCLUSION: This study showed that AOA improved some aspects of cycles at risk of ICSI failure by increasing the fertilization and transferable embryo rates. But blastulation, pregnancy and implantation rates were not improved. The study is limited by its small size and absence of data on cumulative outcomes.

Non-canonical RNA polyadenylation polymerase FAM46C is essential for fastening sperm head and flagellum in mice†.

Family with sequence similarity 46, member C (FAM46C) is a highly conserved non-canonical RNA polyadenylation polymerase that is abundantly expressed in human and mouse testes and is frequently mutated in patients with multiple myeloma. However, its physiological role remains largely unknown. In this study, we found that FAM46C is specifically localized to the manchette of spermatids in mouse testes, a transient microtubule-based structure mainly involved in nuclear shaping and intra-flagellar protein traffic. Gene knockout of FAM46C in mice resulted in male sterility, characterized by the production of headless spermatozoa in testes. Sperm heads were intermittently found in the epididymides of FAM46C knockout mice, but their fertilization ability was severely compromised based on the results of intracytoplasmic sperm injection assays. Interestingly, our RNA-sequencing analyses of FAM46C knockout testes revealed that mRNA levels of only nine genes were significantly altered compared to wild-type ones (q < 0.05). When considering alternate activities for FAM46C, in vitro assays demonstrated that FAM46C does not exhibit protein kinase or AMPylation activity against general substrates. Together, our data show that FAM46C in spermatids is a novel component in fastening the sperm head and flagellum.

A clinical trial on the consistency of bilateral testicular tissue histopathology and Johnsen score: single side or bilateral side biopsy?

To evaluate and compare left and right testicular tissue histopathology and Johnsen score, and to investigate the necessity for bilateral testicular biopsy. We recruited180 patients with non-obstructiveazoospermia (NOA) on testicular biopsy who had undergonetesticular sperm aspiration (TESA). Pathological sections of testicular tissue were diagnosed by specially-assigned doctors, who evaluated pathological findings, determined the Johnsen score and confirmed for the presence or absence of sperm. Sperm positive rates for left and right testicular histopathology were 55.0% and 51.7% respectively, and the proportion of Johnsen scores≥8 for left and right testes were 53.3% and 50.0%, respectively. Cohen kappa values revealed that the identification of sperm in bilateral testicular samples was not consistent and was related to random effects; Optimized cut-off value for bilateral testicular volume was 11ml (Johnsen score ≥8), and optimized cut-off values of E2 on left and right testes were 144.5pmol/L and 133.5 pmol/L (Johnsen score≤7). However, age, serum prolactin (PRL), follicle stimulating hormone (FSH), luteinizing hormone (LH) and total testosterone (TT) levels were not accurate predictors for the existence of testicular sperm. There was nostatistical significance between left and right testicular histopathology in terms of sperm positive rates or Johnsen score; the Johnsen score were caused entirely by random effects and a score from one side could not represent the other side. Therefore, we recommend that both testes need to undergo surgery when NOA patients undergo testicular biopsy or sperm retrieval.

Ran-binding protein M is associated with human spermatogenesis and oogenesis.

The aim of the present study was to explore the underlying mechanism and diagnostic potential of Ran­binding protein M (RanBPM) in human spermatogenesis and oogenesis. RanBPM expression in human testis and ovaries was analysed using polymerase chain reaction (PCR) and western blotting, and immunofluorescence was performed on testis and ovary tissue sections during different developmental stages of spermatogenesis and oogenesis using RanBPM antibodies. Interactions with a variety of functional proteins were also investigated. RanBPM mRNA and protein expression levels were determined by PCR and western blotting in the tissue sections. Results revealed that the mRNA expression levels were highest in the testis followed by the ovary. The RanBPM protein was predominantly localized in the nucleus of germ cells, and the expression levels were highest in pachytene spermatocytes and cells surrounding spermatids in testis tissue. In ovary cells, RanBPM was localized in the nucleus and cytoplasm. In conclusion, the results suggested that RanBPM may have multiple roles in the regulation of germ cell proliferation during human spermatogenesis and oogenesis. This research may provide a novel insight into the underlying molecular mechanism of RanBPM and may have implications for the clinical diagnosis and treatment of human infertility.

Effect of repeated cryopreservation on human embryo developmental potential.

Repeated cryopreservation of surplus embryos from frozen-thawed cycles should occasionally be considered. The purpose of this retrospective cohort study was to evaluate the pregnancy and perinatal outcome of repeated cryopreservation by vitrification of human blastocysts derived from slowly frozen-thawed day 3 embryos. In total, 571 vitrified-warmed blastocyst transfer cycles were investigated. The vitrified-warmed blastocysts were derived from slowly frozen-thawed cleavage embryos (twice-cryopreserved group) or fresh embryos (control group) cultured to the blastocyst stage. Age, body mass index, endometrial thickness, blastocyst developmental rate and number of embryos transferred were not significantly different between twice-cryopreserved and control groups. Clinical pregnancy and implantation rates were also similar. Compared with controls, the miscarriage rate was significantly higher in the twice-cryopreserved group (33.93% versus 19.07%, P = 0.017). This resulted in a significantly lower live birth rate in the twice-cryopreserved group than in controls (29.13% versus 39.18, P = 0.038). No differences were observed in mean gestational age, birthweight and sex ratio of newborns between groups. In conclusion, acceptable clinical pregnancy outcomes may be expected from transfer of twice-cryopreserved human embryos. While the neonatal outcome is not affected, the correlation between the risk of higher pregnancy loss and repeated cryopreservation needs further investigation.

Role of tripartite motif protein 27 as a gametogenesis-related protein in human germ cells.

BACKGROUND: The distribution and functional integrity of members of the tripartite motif (TRIM) protein family are essential for cell proliferation, development and apoptosis, and TRIM proteins have been linked to various cancers. To explore the diagnostic potential and mechanisms of TRIM27 in human spermatogenesis and oogenesis, we analyzed its localization pattern and putative roles in human testes and ovaries. METHODS: TRIM27 mRNA and protein levels in human testes and ovaries were investigated using RT-PCR and western blotting, respectively. TRIM27 was abundantly transcribed in human testes and ovaries, particularly during the early stages of spermatogenesis, and localized in the nuclei of primary spermatocytes. Immunofluorescence also revealed a diffuse distribution in the cytoplasm of round spermatids, and the protein was abundant in ovary tissue during various stages of oogenesis development. RESULTS: TRIM27 mRNA and protein was abundantly transcribed in male and female human germ cells by RT-PCR and western blotting in the human testes followed by the ovary. Immunohistochemical results revealed TRIM27 protein was abundant in the sex body of primary spermatocytes undergoing meiotic prophase during the first cycle of spermatogenesis. Moreover, Trim27 was diffusely localized in the cytoplasm of spermatids and round spermatids. Furthermore, TRIM27 was localized to both the nucleus and cytoplasm of human ovary cells. CONCLUSIONS: TRIM27 as a gametogenesis-related protein could play multiple roles in the regulation of sex body formation and germ cell proliferation during spermatogenesis and oogenesis. The identification and characterization of TRIM27 enhances our understanding of the molecular mechanisms underpinning its functions, and provides insight into its potential role in the pathogenesis of germ cell differentiation and infertility.

Trim27 interacts with Slx2, is associated with meiotic processes during spermatogenesis.

ABSTARCT Formation of the XY body is believed to prevent recombination between X and Y chromosomes during meiosis. We recently demonstrated that SYCP3-like X-linked 2 (Slx2) could be involved in synaptonemal complex formation as well as XY body maintenance during meiosis. In order to further investigate the role and composition of XY body protein complexes in meiotic processes and spermatogenesis, a yeast 2-hybrid screening was performed, and the tripartite motif protein 27(Trim27) was found to interact with Slx2 and co-localized in the XY body. Trim27 has a tripartite motif (TRIM) consisting of a RING finger, B-box and coiled-coil domains, and is a transcriptional regulator that is expressed in various tumor cell lines. In this study, we showed that Slx2 and Trim27 were highly expressed in meiosis of mouse testis. And the Slx2/Trim27 interaction was confirmed in vivo by co-immunoprecipitation and mammalian 2-hybrid interaction assays. Moreover, cytoimmuno localization experiments revealed that Slx2/Trim27 was co-localized to the XY body of spermatocytes during meiosis, and immunohistochemical results revealed co-localization of Trim27 and γ-H2AX in the XY body of primary spermatocytes in the mouse testis. Trim27 may therefore be a transcriptional regulation protein connecting Slx2 and γ-H2AX, thereby promoting the formation of a more potent XY body protein complex in meiotic processes and spermatogenesis. In conclusion, Trim27 connecting Slx2 may regulate meiotic processes in multiple ways by influencing XY body formation and germ cell proliferation during spermatogenesis.

The prevalence of erectile dysfunction among subjects with late-onset hypogonadism: a population-based study in China.

INTRODUCTION: The concurrence of chronic diseases and some well-defined risk factors significantly impacts the prevalence of erectile dysfunction (ED). AIM: To determine whether late-onset hypogonadism (LOH) impacts the prevalence of ED using investigation reproductive health data of middle-aged and aging males in China. METHODS: The reproductive health status of 1498 males, aged 40-69 years, was evaluated using questionnaires of LOH based on the Androgen Deficiency in Aging Males (ADAM) and Aging Male Symptoms scale (AMS), as well as the International Index of Erectile Function-5 (IIEF-5) assessment. The 10th percentile of serum total testosterone (TT) and calculated free testosterone (cFT) levels of controls were set as cut-off levels of AD. The main outcome measures were used to assess the prevalence of LOH and ED according to different subject characteristics. RESULTS: Of the 1472 subjects who completed the questionnaires who supplied hormone measurements, the prevalence of self-reported ED and identified by the IIEF-5 assessment were 11.28% and 77.85%, respectively. The IIEF-5 assessment revealed a prevalence of ED of 55.34%, 88.20%, and 91.77%, respectively, among those aged 40-49, 50-59, and 60-69 years. AD rates of ED subjects were 13.73% and 40.69% according to the TT and cFT cut-off levels. The prevalence of ED among subjects positive for LOH (ADAM+ and AMS+) were 88.81% and 95.80%, respectively. The prevalence of ED among the AD subjects (TT and cFT cut-off levels) with LOH (ADAM+ and AMS+) were 86.67%/81.82%. And the prevalence of ED among clinical LOH subjects (ADAM+ and AMS+) were 89.51%/98.48%. CONCLUSIONS: We found that middle-aged and aging Chinese males were at a relatively high risk of ED. The prevalence of ED among subjects with LOH symptoms was greater than in all recruited subjects. The effect of LOH on the prevalence of ED far outweighed the risk of decreased testosterone levels.

Effect of male body mass index on live-birth sex ratio of singletons after assisted reproduction technology.

OBJECTIVE: To determine the effect of male body mass index (BMI) on the probability of achieving a live birth and the sex ratio of singletons at birth after IVF and intracytoplasmic sperm injection (ICSI) treatment. DESIGN: A retrospective cohort study. SETTING: University-affiliated infertility center. PATIENT(S): Patients seeking infertility treatment who received IVF or ICSI treatment with autologous oocytes from January 2009 to December 2013. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Live-birth sex ratio of singletons at birth stratified by male BMI and adjusted by parental age, parental BMI, type of infertility, parity, embryo culture media, and cause of infertility. RESULT(S): A total of 8,490 couples undergoing IVF or ICSI treatment resulted in 39.12% live births and gave birth to 2,377 live birth singletons and 943 twins. There was no significant difference in the live birth rate between groups stratified by BMI. The probability of live births for overweight and obese groups were not decreased compared with the normal-weight group; similar null findings existed in the IVF and ICSI subgroups. Of note, the sex ratio of offspring in the overweight and obese male groups was significantly higher than in the normal-weight group (1.27 vs. 1.07). Male BMI was significantly associated with sex ratio of singletons after adjusting for confounders. In twins, incidences of twins with male-male infants in the overweight/obese group were not different from the normal-weight group. CONCLUSION(S): Increased male BMI has no effect on live birth success, but has an increased probability of giving birth to male singletons.

Effect of sildenafil on erectile dysfunction and improvement in the quality of sexual life in China: a multi-center study.

BACKGROUND: Erectile dysfunction (ED) is a common medical condition in middle-aged and elderly men; however, large-scale and multi-center epidemiologic studies about the treatment effects on ED in China are lacking. OBJECTIVE: To elucidate the efficacy and safety of a phosphodiesterase type 5 inhibitor (PDE5-i) in the treatment of men with ED in China. METHODS: Patients clinically diagnosed with ED from 53 andrology centers in 15 metropolitan areas in China who were willing to undergo treatment for ED were enrolled in the study. Each participant received 4 weeks of unique PDE5-i treatment, and completed the following forms (International Index of Erectile Function score 5 [IIEF-5], the Erection Hardness Score [EHS], Self-Esteem and Relationship [SEAR], and SF-36 of Health Related Quality of Life). Pre-and post-treatment data were compared using descriptive analysis. RESULTS: A total of 1956 ED patients were included in this study; 1922 patients provided valid questionnaires for analysis. Four weeks of sildenafil treatment was considered effective and safe. Specifically, the IIEF-5 sores (11.30 ± 3.7 vs. 20.02 ± 5.1, P < 0.05), EHS levels (99.1% patients increases to level 3 or 4), and the SEAR scores (32.5 vs. 55.1, P < 0.05) were significantly improved compared to baseline. Sildenafil therapy also significantly improved the satisfaction, enjoyment, and frequency of sexual attempts and sexual activity, as well as physical vigor and mental health scores. CONCLUSION: The present study provides direct evidence regarding the efficacy and safety of sildenafil therapy in a large sample of Chinese men with ED, thus verifying that sildenafil improved the symptoms and quality of sexual life.

Identification and Characterization of Xlr5c as a Novel Nuclear Localization Protein in Mouse Germ Cells.

BACKGROUND: Spermatogenesis is the complex process by which diploid stem cells generate haploid germ cells in gamete production. Members of the Xlr (X-chromosome linked, lymphocyte regulated) superfamily play essential roles in spermatogenesis. The expression, localization and role in spermatogenesis of one such member, Xlr5c, has not been reported previously. METHODOLOGY/PRINCIPAL FINDINGS: Xlr5c mRNA and protein levels in murine testes and other tissues were investigated using RT-PCR and Western blotting. Xlr5c was abundantly transcribed in mouse testes, particularly during the early stages of spermatogenesis and throughout prophase I in the nuclei of spermatocytes. Xlr5c was specifically localized at synaptonemal complexes(SCs) region in preleptotene and pachytene spermatocytes, as was the homologous Xlr protein Sycp3. CONCLUSIONS/SIGNIFICANCE: These results suggest that Xlr5c was abundantly transcribed in germ cells, localized at SCs region, where it may play a potential role during the early stages of spermatogenesis. Identification and characterization of this novel testis protein may offer a new perspective for understanding of the molecular mechanisms involved in germ cell differentiation.

Effect of embryo culture media on percentage of males at birth.

STUDY QUESTION: Does embryo culture medium influence the percentage of males at birth? SUMMARY ANSWER: The percentage of males delivered after ICSI cycles using G5™ medium was statistically significantly higher than after cycles where Global, G5™ PLUS, and Quinn's Advantage Media were used. WHAT IS KNOWN ALREADY: Male and female embryos have different physiologies during preimplantation development. Manipulating the energy substrate and adding growth factors have a differential impact on the development of male and female embryos. STUDY DESIGN, SIZE AND DURATION: This was a retrospective analysis of the percentage of males at birth, and included 4411 singletons born from fresh embryo transfer cycles between January 2011 and August 2013 at the Center for Reproductive Medicine of Third Hospital Peking University. PARTICIPANTS/MATERIALS, SETTING, AND METHODS: Only singleton gestations were included. Participants were excluded if preimplantation genetic diagnosis, donor oocytes and donor sperm were used. The database between January 2011 and August 2013 was searched with unique medical record number, all patients were present in the database with only one cycle. Demographics, cycle characteristics and the percentage of male babies in the four culture media groups were compared with analysis of variance or χ(2) tests. Multivariable logistic regression was done to determine the association between the sex at birth and culture media after adjusting for other confounding factors, including parental age, parental BMI, type of infertility, parity, number of embryos transferred, number of early gestational sacs, cycles with testicular sperm aspiration (TESA)/percutaneous epididymal sperm aspiration (PESA)/testicular sperm extraction (TESE), number of oocytes retrieved, cycles with blastocyst transfers, and gestational age within ICSI group. MAIN RESULTS AND THE ROLE OF CHANCE: Within the IVF group, the percentage of males at birth for G5™, Global, Quinn's and G5™ PLUS media were comparable (P > 0.05); however, within the ICSI group, the percentage of male babies in cycles using G5™(56.1%) was statistically significantly higher than in cycles that used Global (47.2%; P = 0.003), G5™ PLUS (47.7%; P = 0.005) or Quinn's media (45.0%; P = 0.009). There were no statistically significant differences in the percentage of males at birth between cycles that used Global, G5™ PLUS and Quinn's media (P > 0.05). Multivariable logistic regression indicated that culture media (G5™ versus Global, G5™ PLUS, and Quinn's) were significantly associated with the sex at birth (P = 0.008) after adjusting for parental age, parental BMI, type of infertility, parity, number of embryos transferred, number of early gestational sacs, cycles with TESA/PESA/TESE, number of oocytes retrieved, cycles with blastocyst transfers, and gestational age. LIMITATIONS AND REASONS FOR CAUTION: This study was not a randomized controlled trial and allocation of treatment cycles over the four media was not completely at random. Cigarette smoking was not included in the current study because this confounding factor was not registered in our database. Moreover, intra-variability of sperm selection between the five embryologists may directly affect the percentage of males. WIDER IMPLICATIONS OF THESE FINDINGS: Our study suggests that human embryogenesis responds differently to G5™, Global, G5™ PLUS and Quinn's Advantage Medium. This finding can be generalized to other commercial culture media. STUDY FUNDING/COMPETING INTERESTS: National Natural Science Foundation of China for Young Scholars (81300483 and 81200466). The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: Not applicable.

miRNA-mediated regulation of heat shock proteins in human ejaculated spermatozoa.

BACKGROUND/AIM: Heat shock proteins (HSPs) are expressed in human spermatozoa and play a role in sperm function. MicroRNAs (miRNAs) regulate gene expression, and the possible involvement of miRNAs in the regulation of HSP gene expression in sperm was investigated in this study. MATERIALS AND METHODS: miRNAs differentially expressed in 8 copies of an oligoasthenozoospermic semen group (OA) were identified by comparison with a normal male semen control group (NC) using microarray technology. Potential targets of HSP proteins among the differentially expressed miRNAs were further investigated. Results: HSP40, HSP60, HSP70, and HSP90 were all found to be expressed in human ejaculated spermatozoa. A total of 32 miRNAs showed significant differences in expression between the OA and NC groups. Ten of these miRNAs encoded potential targets of HSPs. CONCLUSION: These results show that specific miRNAs are expressed in human ejaculated spermatozoa. These miRNAs appear to be involved in regulating the expression of HSP40, HSP70, and HSP90, and this in turn affects sperm function.

Correlation between body mass index and semen quality in male infertility patients.

BACKGROUND/AIM: The question of whether body mass index (BMI) affects semen quality and male fertility is controversial. The purpose of this research was to determine whether there is a correlation between BMI and semen analysis parameters. MATERIALS AND METHODS: A total of 617 male infertility patients were recruited and separated into 3 groups according to BMI values as follows: normal weight group (n = 334), overweight group (n = 220), and obese group (n = 63). Height and weight were measured and a routine semen analysis was performed for all patients. RESULTS: Significant differences existed in BMI, age, and sperm motility (progressive motility) among the 3 groups. BMI and abstinence period were negatively correlated with sperm motility (P < 0.05 and P < 0.01), although they did not correlate with semen volume, total sperm number, concentration, and rate of sperm with normal morphology (P > 0.05). Abstinence, BMI, and age had a linear correlation with sperm motility (P < 0.01) in that order of influence. CONCLUSION: Sperm motility, an important semen parameter with respect to male fertility, is reduced in men with increased BMI, and BMI is one of the risk factors that influence semen quality.

Up-regulation of heme oxygenase-1 expression modulates reactive oxygen species level during the cryopreservation of human seminiferous tubules.

OBJECTIVE: To study the effect of freezing techniques and to optimize a method for trace amounts of testicular spermatozoa from biopsed seminiferous tubules. The level of reactive oxygen species (ROS) and the gene expression of heme oxygenase-1 was evaluated. DESIGN: Prospective experimental study. SETTING: University-based laboratory. PATIENT(S): Eighteen adults with male fator infertility underwent testicular biopsy surgery. INTERVENTION(S): Seminiferous tubular fragments from each man were evenly allocated to three groups: fresh control, slow freezing, and vitrifiaction groups. The morphology and ROS levels before and after freezing were evaluated for seminiferous tubular fragments. The expression of heme oxygenase-1 (HO-1) at both the transcriptional and protein levels was determined. MAIN OUTCOME MEASURE(S): The morphology was analyzed by light microscopy. The ROS levels were measured with ELISA. The proliferation and differentiation were evaluated by immunohistochemistry, and the expression of HO-1 was evaluated using a real-time polymerase chain reaction (PCR) and Western blotting. RESULT(S): Decreased ROS levels and increased HO-1 expression at the transcriptional and protein levels were observed after thawing the human seminiferous tubules. The ROS level was negatively correlated with HO-1 expression. Slow freezing was more effective than vitrification in terms of HO-1 up-regulation and ROS alteration. CONCLUSION(S): Based on our study, the slow freezing technique was more effective compared with the vitrification method.