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Asthenozoospermia (ASZ) is a condition characterized by reduced forward motility of spermatozoa affecting approximately 19% of infertile men. A kinase anchor protein 4 (AKAP4) is an X-linked testis-specific gene and plays a major role in sperm motility and flagella formation. However, few studies have reported its association with ASZ. Here, we sequenced for exonic mutations of human AKAP4 gene by high-fidelity PCR/Sanger sequencing in peripheral blood samples from 150 ASZ patients and 150 fertile men. We reported the identification of three novel hemizygous mutations unique to four ASZ patients, including one patient carrying missense mutation c.454T>C (p.S152P), two patient carrying synonymous mutation c.1173T>C (p.H391H), and one patient carrying synonymous mutation c.2007 A>G (p.R669R). The p.S152P mutation was located in a precursor pro-polypeptide domain of AKAP4 protein, which was predicted to be damaging by SIFT and PolyPhen-2 and could cause the protein accumulation in the cytoplasm of COS-7 cells. The mature protein of AKAP4 was absent in spermatozoa of ASZ patient harboring AKAP4 p.S152P mutation. Further in vitro cellular assays showed that reactive oxygen species (ROS), malondialdehyde (MDA), myeloperoxidase (MPO) levels, and apoptotic cells were increased in GC2-spd cells by AKAP4 p.S152P mutant protein, whereas superoxide dismutase (SOD) level was decreased. AKAP4 p.H391H and p.R669R mutant proteins were coimmunoprecipitated with ribonuclease T2 (RNASET2) protein in GC2-spd cells, whereas no interaction between the AKAP4 p.S152P mutant protein and RNASET2 protein was observed. In addition, AKAP4 p.S152P mutant protein could decrease the activity of PKA/PI3K signaling. Overall, our study identifies a novel AKAP4 p.S152P mutation is associated with ASZ probably through affecting oxidative stress and cell apoptosis by regulating the interaction with RNASET2 and the activity of the PKA/PI3K signaling pathway.

STUDY QUESTION Can a counselling tool be developed for couples with different types of azoospermia to predict the probability of clinical pregnancy in ICSI after surgical sperm retrieval? SUMMARY ANSWER A prediction model for clinical pregnancy in ICSI after surgical sperm retrieval in different types of azoospermia was created and clinical type of azoospermia, testicular size, male FSH, male LH, male testosterone, female age, female antral follicle count (AFC) and female anti-Müllerian hormone (AMH) were used as predictors. WHAT IS KNOWN ALREADY Prediction models are used frequently to predict treatment success in reproductive medicine; however, there are few prediction models only for azoospermia couples who intend to conceive through surgical sperm retrieval and ICSI. Furthermore, no specific clinical types of azoospermia have been reported as predictors. STUDY DESIGN, SIZE, DURATION A cohort study of 453 couples undergoing ICSI was conducted between 2016 and 2019 in an academic teaching hospital. PARTICIPANTS/MATERIALS, SETTING, METHODS Couples undergoing ICSI with surgically retrieved sperm were included, with 302 couples included in the development set and 151 couples included in the validation set. We constructed a prediction model using multivariable logistic regression analysis. The internal validation was based on discrimination and calibration. MAIN RESULTS AND THE ROLE OF CHANCE We found that for male patients involved in our model, different clinical types of azoospermia are associated with different clinical pregnancy outcomes after ICSI. Considering the clinical type of azoospermia, larger testicular volume and higher levels of FSH, LH and testosterone in the body are associated with higher clinical pregnancy success rates. For women involved in our model, younger age and higher AFC and AMH levels are associated with higher clinical pregnancy success rates. In the development set, the AUC was 0.891 (95% CI 0.849–0.934), indicating that the model had good discrimination. The slope of the calibration plot was 1.020 (95% CI 0.899–1.142) and the intercept of the calibration plot was −0.015 (95% CI −0.112 to 0.082), indicating that the model was well-calibrated. From the validation set, the model had good discriminative capacity (AUC 0.866, 95% CI 0.808–0.924) and calibrated well, with a slope of 1.015 (95% CI 0.790–1.239) and an intercept of −0.014 (95% CI −0.180 to 0.152) in the calibration plot. LIMITATIONS, REASONS FOR CAUTION We found that BMI was not an effective indicator for predicting clinical pregnancy, which was inconsistent with some other studies. We lacked data about the predictors that reflected sperm characteristics, therefore, we included the clinical type of azoospermia instead as a predictor because it is related to sperm quality. We found that almost all patients did not have regular alcohol consumption, so we did not use alcohol consumption as a possible predictor, because of insufficient data on drinking habits. We acknowledge that our development set might not be a perfect representation of the population, although this is a common limitation that researchers often encounter when developing prediction models. The number of non-obstructive azoospermia patients that we could include in the analysis was limited due to the success rate of surgical sperm retrieval, although this did not affect the establishment and validation of our model. Finally, this prediction model was developed in a single centre. Although our model was validated in an independent dataset from our centre, validation for different clinical populations belonging to other centres is required before it can be exported. WIDER IMPLICATIONS OF THE FINDINGS This model enables the differentiation between couples with a low or high chance of reaching a clinical pregnancy through ICSI after surgical sperm retrieval. As such it can provide couples dealing with azoospermia a new approach to help them choose between surgical sperm retrieval with ICSI and the use of donor sperm. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by a grant from the National Natural Science Foundations of China (81501246 and 81501020 and 81671443). The authors declare no competing interest. TRIAL REGISTRATION NUMBER N/A.

To investigate the expressions of Th1/Th2 cytokines and autoantibodies in the serum of the patients with idiopathic oligoasthenozoospermia (IO) and their significance.From November 2017 to April 2020, we examined the levels of Th1/Th2 cytokines (IL-2, IL-4, IL-10, IL-21, TNF-α, IFN-γ) and the expressions of AhCGAb, AsAb and AcAb in the serum of 48 infertile men with mild or moderate IO, 48 with severe IO and another 72 males with normal semen parameters by ELISA. We compared the results of detection among the three groups and analyzed them with the logistic regression model.Compared with the normal controls, the patients with mild or moderate IO showed significant increases in the levels of IL-10, IL-21 and IFN-γ and the expressions of AhCGAb, AsAb and AcAb (P0.05), and so were those of the severe IO group in the levels of all the six cytokines and the expressions of the three autoantibodies (P0.05). The levels of IL-2, IL-4, IL-21 and TNF-α and the expressions of AhCGAb and AsAb were even higher in the patients with severe IO than in those with mild or moderate IO (P0.05). Multivariate logistic regression analysis showed that the increased levels of IL-21/IL-10 (OR = 1.694, 95% CI: 0.319－4.035, P0.05) and positive expressions of AhCGAb (OR = 4.357, 95% CI: 1.204－9.426, P0.05) and AsAb (OR = 2.135, 95% CI: 1.902－5.429, P0.05) were the risk factors for IO.The levels of the cytokines IL-2, IL-4, IL-10, IL-21, TNF-α and IFN-γ and the expressions of the autoantibodies AhCGAb, AsAb and AoAb are significantly higher in IO patients than in normal healthy males. Quantitative analysis of cytokines and autoantibodies in the serum of IO patients may provide some valuable information for studies of the pathogenesis of male infertility.

To investigate the effects of Compound Amino Acid Capsules (CAAC) for obstructive azoospermia (OA) patients undergoing percutaneous epididymal sperm aspiration (PESA) and ICSI.We retrospectively studied 134 cycles of PESA-ICSI in OA males, who were divided into a CAAC group (n = 64) and a control group (n = 70), the former, aged (32.1 ± 5.7) years, treated with CAAC for 3 months before PESA-ICSI while the latter, aged (32.3 ± 4.5) years, left untreated. We compared the rates fertilization, cleavage, high-quality embryos and blastocyst formation between the two groups of patients.Compared with the control, the CAAC group achieved significantly higher rates of high-quality embryos (［67.4 ± 13.8］% vs ［74.3 ± 12.6］%, P0.05) and blastocyst formation (［60.3±14.2］% vs ［66.8±14.6］%, P0.05), but no statistically significant differences were observed between the two groups in the ages of the males and females, number of oocytes retrieved, or the rates of fertilization and cleavage.Medication of Compound Amino Acid Capsules can effectively improve the rates of high-quality embryos and blastocyst formation in obstructive azoospermia patients undergoing PESA-ICSI.

To investigate the efficiency of microdissection testicular sperm extraction (micro-TESE) in male patients with nonmosaic Klinefelter's syndrome (NMKS), the outcomes of intracytoplasmic sperm injection (ICSI) in their wives, and the possible predictors of clinical pregnancy.Forty-nine males with NMKS underwent micro-TESE in our hospital from July 2016 to November 2018. We compared the age, reproductive hormone levels, and testis volume of the patients between the sperm-positive and -negative groups. We performed ICSI for the wives of the sperm-positive patients, recorded the numbers of pregnancies and births, compared the age, reproductive hormone levels and number of mature oocytes between the successful and failed ICSI groups, and analyzed the possible predictors of the results of micro-TESE and outcomes of ICSI.The 49 patients were aged (28.20 ± 3.52) years, all diagnosed as with 47,XXY nonmosaicism by karyotype analysis, with a testis volume of (2.95 ± 0.84) ml, a serum FSH content of (42.42 ± 14.37) IU/L, a serum LH level of (22.50 ± 8.64) IU/L, and a serum T level of (6.64 ± 4.13) nmol/L. Sperm were obtained from 32 of the patients, with a sperm retrieval rate (SRR) of 65.31%, and the wives (aged ［26.79 ± 2.97］ years) of 29 of the sperm-positive males underwent ICSI, achieving a fertilization rate of (48.14 ± 27.33)%, an available embryo rate of (63.71 ± 28.90)%, a pregnancy rate of 48.28% (14/29), and a birth rate of 24.14% (7/29) up to the present time, with 7 cases awaiting delivery. The 2 cases failing to achieve pregnancy were waiting for transplantation of the frozen embryos. Logistic regression analysis showed that the preoperative serum T level of the NMKS patients had a significant predictive value for the pregnancy rate (AUC = 0.832, cut-off value = 5.17 nmol/L, P = 0.015), but not the other factors for either the SRR or the pregnancy rate.Sperm can be retrieved from over 60% of the NMKS patients undergoing micro-TESE, and some of them can achieve pregnancy and have their own children by ICSI. Moreover, those with a preoperative serum T level of5.17 nmol/L are very likely to achieve clinical pregnancy after successful sperm retrieval.

To compare the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (micro-TESE) and the outcomes of intracytoplasmic sperm injection (ICSI) among different etiological types of non-obstructive azoospermia (NOA).We retrospectively analyzed the clinical data on 155 cases of NOA undergoing micro-TESE in our hospital from September 2016 to December 2017, which were classified into three types according to etiological factors: congenital NOA (n = 49), acquired NOA (n = 15) and idiopathic NOA (n = 91). We compared the age, testis volume, levels of reproductive hormones, ultrasonographic manifestations, and SRR of micro-TESE among the three groups of patients. We also recorded and analyzed the rates of fertilization, available embryos and clinical pregnancy in the spouses of the patients included for successful sperm retrieval in micro-TESE.The testis volume was significantly lower in the congenital than in the acquired and idiopathic NOA groups (［6.4 ± 5.0］ vs ［10.2 ± 2.0］ and ［9.9 ± 3.2］ ml, P0.05), while the LH level was markedly higher in the former group than in the latter two (［15.2 ± 10.1］ vs ［9.1 ± 6.5］ and ［7.8 ± 3.5］ mIU/ml, P0.05), and so was the T level in the idiopathic than in the congenital NOA group (［11.8 ± 4.8］ vs ［8.9 ± 4.5］ nmol /L, P0.05). The SRRs of micro-TESE in the congenital, acquired and idiopathic NOA patients were 73.5% (36/49), 100% (15/15), and 24.2% (22/91) respectively, with statistically significant differences among the three groups (P0.05). The fertilization rate after ICSI was remarkably higher in the acquired than in the congenital and idiopathic NOA groups (［73.1±23.3］% vs ［48.9±21.7］% and ［52.6±22.7］%, P0.05). There were no statistically significant differences among the three groups in the rates of embryo utilization and clinical pregnancy.The sperm retrieval rate of micro-TESE and the rates of fertilization, embryo utilization and clinical pregnancy after ICSI were the highest in the acquired NOA but the lowest in the idiopathic NOA patients.

This study aimed to investigate the relationships between environmental exposure to metals/metalloids and semen quality, sperm apoptosis and DNA integrity using the metal/metalloids levels in seminal plasma as biomarkers. We determined 18 metals/metalloids in seminal plasma using an inductively coupled plasma-mass spectrometry among 746 men recruited from a reproductive medicine center. Associations of these metals/metalloids with semen quality (n = 746), sperm apoptosis (n = 331) and DNA integrity (n = 404) were evaluated using multivariate linear and logistic regression models. After accounting for multiple comparisons and confounders, seminal plasma arsenic (As) quartiles were negatively associated with progressive and total sperm motility using multivariable linear regression analysis, which were in accordance with the trends for increased odds ratios (ORs) for below-reference semen quality parameters in the logistic models. We also found inverse correlations between cadmium (Cd) quartiles and progressive and total sperm motility, whereas positive correlations between zinc (Zn) quartiles and sperm concentration, between copper (Cu) and As quartiles and the percentage of tail DNA, between As and selenium (Se) quartiles and tail extent and tail distributed moment, and between tin (Sn) categories and the percentage of necrotic spermatozoa (all Ptrend

Background: Exposure to metals, including essential and nonessential elements, has been linked to male reproductive health in animals. However, findings from human studies are inconsistent. Objective: To examine the associations between metal exposure at environmental levels and semen quality in a Chinese population by performing repeated measurements of urinary metals to estimate individual-level exposure. Methods: From March to June 2013, 1052 men seeking semen evaluation were recruited from the Reproductive Center of Tongji Hospital in Wuhan, China. Each man provided one semen sample and two urine sample. Semen quality parameters and urinary levels of 18 metals were determined. Associations between the urinary metal levels and semen quality parameters were assessed using confounder adjusted linear and logistic regressions. Restricted cubic spline analysis was performed to assess dose-response relationships between continuous metal measurements and outcomes. Results: Urinary levels of cadmium were significantly inversely associated with progressive sperm motility and total motility (both P

Toxicological studies showed that trihalomethanes (THMs), the most abundant classes of disinfection by-products (DBPs) in drinking water, impaired male reproductive health, but epidemiological evidence is limited and inconsistent. This study aimed to examine the associations of baseline blood THMs with semen parameters and serum total testosterone in a Chinese population. We recruited 401 men seeking semen examination from the Reproductive Center of Tongji Hospital in Wuhan, China between April 2011 and May 2012. Baseline blood concentrations of THMs, including chloroform (TCM), bromodichloromethane (BDCM), dibromochloromethane (DBCM), and bromoform (TBM) were measured using SPME-GC/ECD method. Semen quality and serum total testosterone were analyzed. Multivariable linear regressions were used to assess the associations of baseline blood THM concentrations with semen parameters and serum total testosterone levels. We found that baseline blood THM concentrations were not associated with decrements in sperm motility, sperm straight-line and curvilinear velocity. However, moderate levels of BDCM (β=−0.13 million; 95% CI: −0.22, −0.03) and DBCM (β=−4.74%; 95% CI: −8.07, −1.42) were associated with decreased sperm count and declined sperm linearity compared with low levels, respectively. Suggestive dose–response relationships were also observed between elevated blood TCM or ∑ THMs (sum of TCM, BDCM, DBCM and TBM) concentration and decreased sperm concentration (both p for trend=0.07), and between elevated blood DBCM concentration and decreased serum total testosterone (p for trend=0.07). Our results indicate that elevated THM exposure may lead to decreased sperm concentration and serum total testosterone. However, the effects of THM exposure on male reproductive health still warrant further studies in humans. Keywords: Blood trihalomethanes, Disinfection by-products, Human, Male reproductive health, Semen quality, Testosterone

Background: Exposure to metals, including essential and nonessential elements, is widespread and may be associated with male reproductive health. Objective: To examine whether environmental exposure to metals contributes to reproductive hormone changes, spermatozoa apoptosis and sperm DNA damage in a Chinese population. Methods: Eighteen metals (aluminum, arsenic, antimony, chromium, cobalt, copper, cadmium, iron, lead, manganese, molybdenum, nickel, selenium, tin, tungsten, thallium, uranium and zinc) were analyzed in two urine samples collected a few hours apart from male partners of couples attending an infertility clinic. Multivariable linear regression models were used to assess the cross-sectional associations of average urinary metal levels with serum hormones (n = 511), spermatozoa apoptosis measures (n = 460) and sperm DNA damage parameters (n = 516). Results: We found significant inverse dose-dependent trends of urinary tin quartiles with total testosterone (T), and tin, nickel, zinc and molybdenum with the ratio of total T to luteinizing hormone (total T/LH ratio) (all Ptrend

To assess the influence of male age on the outcome of oocyte donation cycles. A total 103 oocyte donation cycles of 70 couples (male aged 26 to 57) were examined, all of which were performed with conventional in vitro fertilization using fresh ejaculation sperm. Main outcome measures were fertilization rate, clinical pregnancy, live birth rates and pregnancy loss. A total 122 cryopreserved embryo transfer were performed, resulting in 34 cycles resulted in clinical pregnancy and 27 live births. No significant correlation was found between male age and fertilization rate. No significant difference was found in male age between the patients who achieved clinical pregnancy and live birth and those who did not. All the pregnancy loss occurred in cycles where the male was older than 37, however, when the cycles were divided into two groups according to whether or not male age older than 37, no statistically significant difference was not found in pregnancy loss rate. Aging of the male has no significant impact on fertilization, pregnancy or live birth in oocyte donation cycles, but may be associated with pregnancy loss.

Previous studies inconsistently suggest that assisted reproduction technology (ART) may increase the risk of birth defects in children.Live birth infants, conceived by in vitro fertilization fresh embryo transfer (IVF), intracytoplasmic sperm injection fresh embryo transfer (ICSI), or frozen-thawed embryo transfer (FET) in Reproductive Center of Tongji Hospital (Wuhan, China) between 1997 and 2008, were followed up at birth and after 3 years. Preterm pregnancy, multiple pregnancy, sex ratio (male/female), congenital malformation were compared.A total of 4,236 children were born after ART (IVF 2,543, ICSI 908, FET 785). Compared with IVF, the rate of preterm pregnancy and sex ratio in ICSI were lower (p0.05); the rate of multiple pregnancy in ICSI and FET were all lower than IVF (p0.05). Congenital defects were comparable in all groups at birth. In total, 2,908 children participated in the second follow-up from 34 months to 60 months with an average of 40 months, and the cases of birth defects had doubled (3 years: 5.16%, birth: 2.22%). The birth defect rate in boys conceived through ICSI was significantly higher than the IVF group after 3-year follow-up (ICSI boys: 8.62%, IVF boys: 5.21% [p0.05]), even though there was no significant difference at birth.Compared with IVF, FET may not increase risk of birth defects. Children conceived through ICSI, especially males, had higher rates of congenital malformations that were inapparent at birth. So longitudinal monitoring may provide insights into the risks of ART.

Couples rarely undergo a karyotype evaluation prior to either natural conception or receiving assisted reproductive technology (ART) if there is no history of abnormal pregnancy or other indicative signs. However, many chromosomal anomalies are not clinically obvious and do not preclude fertility nor do they cause miscarriage. Carriers may then conceive and produce a genetically abnormal child, and the defect is not detected until after birth.

To explore the effects of sperm chromatin structure abnormalities on the outcome of in vitro fertilization and embryo transfer (IVF-ET).Sperm DNA fragmentation and chromatin packaging defects were assessed in 136 couples undergoing IVF-ET because of infertility. The relationship between sperm DNA fragmentation, chromatin packaging defects and fertilization rate and clinical pregnancy was evaluated.Both sperm DNA fragmentation and chromatin packaging defect had a negative correlation with fertilization rate (r=-0.198, P0.05, and r=-0.389, P0.01, respectively). Both parameters were higher in couples who failed to achieve pregnancy than those who achieved clinical pregnancy (10.74% vs. 5.40%, P0.01 and 23.58% vs. 11.83%, P0.01, respectively).Abnormality of sperm chromatin structure is one of the reasons for IVF-ET failure. Examination of sperm chromatin structure is helpful in predicting the risk of IVF-ET failure and optimizing treatment of infertility.

To report the outcome of intracytoplasmic sperm injection (ICSI) cycles of an oligozoospermic man with reciprocal translocation.Two ICSI cycles were given to a 29-year-old man with severe oligozoospermia and reciprocal translocation t(18;21)(p11;q21) and his wife. In the first cycle, no sperm were found in his semen and all 15 oocytes retrieved were donated to another infertile couple suffering from oligozoospermia and premature ovarian failure. In the second cycle, sperm from the husband's ejaculate were used to fertilize 13 retrieved oocytes.Eleven embryos were acquired and a healthy girl was born in the oocyte-donated ICSI cycle. In the second cycle, 6 embryos were acquired and only biochemical pregnancy was achieved after three times of embryo transfer.By the unusual oocyte-donating ICSI procedures, impact of the new reported reciprocal translocation t (18; 21) (p11; q21) on male fertility and embryo development was suggested.

Conflicting conclusions present in earlier studies on the relationship between sperm chromatin anomalies and outcome of conventional in vitro fertilization (IVF) may result from confounding by poorly controlled external factors. We examined the relationship of sperm chromatin parameters and outcome of conventional IVF while strictly controlling external factors.