Sperm retrieval rates and clinical outcomes for patients with different causes of azoospermia who undergo microdissection testicular sperm extraction-intracytoplasmic sperm injection

The aim of our study was to compare the sperm retrieval rates(SRRs)and clinical outcomes of patients with different causes of azoospermia who underwent microdissection testicular sperm extraction-intracytoplasmic sperm injection(micro-TESE-ICSI).We conducted a retrospective study at the Reproductive Medicine Center of Peking University Third Hospital in Beijing,China,from January 2014 to December 2017.This study examined 769 patients with nonobstructive azoospermia who underwent 347 cycles of micro-TESE-ICSI.Patients with azoospermia were classified into Group A(Klinefelter syndrome,n=284,125 cycles),Group B(azoospermia Y chromosome factor c[AZFc]microdeletion,n=91,64 cycles),Group C(cryptorchidism,n=52,39 cycles),Group D(previous mumps and bilateral orchitis,n=23,23 cycles),and Group E(idiopathic azoospermia,n=319,96 cycles).Clinical characteristics,SRR,embryonic development,and pregnancy outcomes of the patients were compared between all groups.Patients in Group D had the highest and most successful SRR.The average SRR for all patients was 46.0%.The rates of clinical pregnancy,implantation,and live birth in Group D were 78.3%,65.0%,and 74.0%,respectively,which were higher than those in all other groups(P<0.05).Group B patients had the lowest clinical pregnancy,implantation,and live birth rates of all groups(P<0.05).No differences were found in the miscarriage rate or birth defects among the groups(P>0.05).Patients with orchitis had the highest SRR and best clinical outcomes.Although AZFc microdeletion patients had a higher SRR,their clinical outcomes were worse.

Altered microRNA profiles of testicular biopsies from patients with nonobstructive azoospermia

Many studies have shown that microRNAs(miRNAs)play vital roles during the spermatogenesis.However,little is known about the altered miRNA profiles of testicular tissues in nonobstructive azoospermia(NOA).Using microarray technology,the miRNA expression profiles of testicular biopsies from patients with NOA and of normal testicular tissues were determined.Bioinformatics analyses were conducted to predict the enriched biological processes and functions of identified miRNAs.The microarray data were validated by quantitative reverse transcriptase polymerase chain reaction(qRT-PCR),the results of which were then validated with a larger sample size.Correlations between the miRNA expression levels and clinical characteristics were analyzed.Receiver operating characteristic(ROC)curve analysis was used to evaluate the diagnostic ability of miRNAs for azoospermia.Hierarchical clustering showed that 129 miRNAs were significantly differentially expressed between the NOA and control groups.Bioinformatics analysis indicated that the differentially expressed miRNAs were involved in spermatogenesis,cell cycle,and mitotic prometaphase.In the subsequent qRT-PCR assays,the selected miRNA expression levels were consistent with the microarray results,and similar validated results were obtained with a larger sample size.Some clinical characteristics were significantly associated with the expression of certain miRNAs.In particular,we identified a combination of two miRNAs(miR-10b-3p and miR-34b-5p)that could serve as a predictive biomarker of azoospermia.This study provides altered miRNA profiles of testicular biopsies from NOA patients and examines the roles of miRNAs in spermatogenesis.These profiles may be useful for predicting and diagnosing the presence of testicular sperm in individuals with azoospermia.

Clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection with fresh or cryopreserved sperm in patients with nonobstructive azoospermia

We performed this study to evaluate the clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection(micro-TESE-ICSI)treatment that used fresh or cryopreserved sperm in patients with nonobstructive azoospermia(NOA).A total of 338 NOA patients with 344 consecutive cycles received treatment in the reproductive medicine center of Peking University Third Hospital in Beijing,China,from January 2014 to December 2017.Fresh oocytes and fresh sperm were used in 222 patients with 234 cycles(Group A).Fresh oocytes and cryopreserved sperm were used in 116 patients with 110 cycles(Group B).We compared patient characteristics,embryonic development,and pregnancy outcomes between Groups A and B.There was no statistical difference in the patient characteristics,and no differences were observed with fertilization or quality embryo rates between Groups A and B.The rates of clinical pregnancy and live birth were both higher for Group A than those for Group B(both P<0.05).In conclusion,fresh testicular sperm appears to produce better ICSI outcomes than cryopreserved testicular sperm in patients with NOA.

Development of a predictive model for increasing sperm retrieval sucpess by microdissection testicular sperm extraction in patients with nonobstructive azoospermia

Microdissection testicular sperm extraction(micro-TESE)is widely used to treat nonobstructive azoospermia.However,a good prediction model is required to anticipate a successful sperm retrieval rate before performing micro-TESE.This retrospective study analyzed the clinical records of 200 nonobstructive azoospermia patients between January 2021 and December 2021.The backward method was used to perform binary logistic regression analysis and identify factors that predicted a successful micro-TESE sperm retrieval.The prediction model was constructed using acquired regression coefficients,and its predictive performance was assessed using the receiver operating characteristic curve.In all,67 patients(sperm retrieval rate:33.5%)underwent successful micro-TESE.Follicle-stimulating hormone,anti-Miillerian hormone,and inhibin B levels varied significantly between patients who underwent successful and unsuccessful micro-TESE.Binary logistic regression analysis yielded the following six predictors:anti-Mullerian hormone(odds ratio[OR]=0.902,95%confidence interval[Cl]:0.821-0.990),inhibin B(OR=1.012,95%Cl:1.001-1.024),Klinefelter’s syndrome(OR=0.022,95%Cl:0.002-0.243),Y chromosome microdeletion(OR=0.050,95%Cl:0.005-0.504),cryptorchidism with orchiopexy(OR=0.085,95%Cl:0.008-0.929),and idiopathic nonobstructive azoospermia(OR=0.031,95%Cl:0.003-0.277).The prediction model had an area under the curve of 0.720(95%Cl:0.645-0.794),sensitivity of 65.7%,specificity of 72.2%,Youden index of 0.379,and cut-off value of 0.305 overall,indicating good predictive value and accuracy.This model can assist clinicians and nonobstructive azoospermia patients in decision-making and avoiding negative micro-TESE results.

Microdeletions and vertical transmission of the Y-chromosome azoospermia factor region

Spermatogenesis is regulated by several Y chromosome-specific genes located in a specific region of the long arm of the Y chromosome,the azoospermia factor region(AZF).AZF microdeletions are the main structural chromosomal abnormalities that cause male infertility.Assisted reproductive technology(ART)has been used to overcome natural fertilization barriers,allowing infertile couples to have children.However,these techniques increase the risk of vertical transmission of genetic defects.Despite widespread awareness of AZF microdeletions,the occurrence of de novo deletions and overexpression,as well as the expansion of AZF microdeletion vertical transmission,remains unknown.This review summarizes the mechanism of AZF microdeletion and the function of the candidate genes in the AZF region and their corresponding clinical phenotypes.Moreover,vertical transmission cases of AZF microdeletions,the impact of vertical inheritance on male fertility,and the prospective direction of research in this field are also outlined.

Sperm Donors in Shanghai,China:A Study of Motivations,Characteristics,and Semen Parameters of Actual Sperm Donors

Objective:To identify the sociodemographic characteristics,motivations,and semen parameters of sperm donors in Shanghai,China.Methods:The participants were sperm donors associated with the Human Sperm Bank of Fudan University in Shanghai,China.Among the 334 sperm donors that applied for participation,329 completed the survey process.The responses obtained in the questionnaire and face-to-face interviews were used to investigate the donor motivations and characteristics,and the semen quality was examined to identify the sperm parameter.Results:In terms of the sociodemographic characteristics,an altruistic donor in this study was aged between 26 and 30 years,was single,did not have a child,had a college or undergraduate education level,was of the Han ethnicity,and worked full time.The strongest motivation highlighted by sperm donors was a donation for altruistic(26.4%,n=87)reasons.The second-highest rated motivation was curiosity(20.7%,n=68),followed by a desire to procreate(17.9%,n=59)."Complimentary body checks"(14.3%,n=47)and"financial incentives"(14.7%,n=47)were regarded as less important.The average semen parameters of the 329 donors were as follows:the semen volume was 3.39±1.21 mL,the semen concentration was 82.75×10^(6)/mL,the progressive motility rate(PR%)was 63.77%±3.13%,the total motility rate was 66.26%,the total progressive motile count was 158.31%±54.43×10^(6)/mL,and the round cell concentration was 0.38±0.51×10^(6)/mL.The PR%of the procreation motivation group was significantly higher than that of the other motivation groups(P<0.05).Conclusions:Sperm donors in Shanghai,China,are altruistic about their donation,although curiosity is also a key motivator.In addition,the decisions of donors are culturally influenced,and the motivation to procreate may influence the PR%.

Nanotechnology-assisted adipose-derived stem cell （ADSC） therapy for erectile dysfunction of cavernous nerve injury： In vivo cell tracking, optimized injection dosage, and functional evaluation

Stem cell therapy is a potentially promising option for erectile dysfunction; however, its risk of tumorigenicity is a clinical hurdle and the risk is positively related to the number of injected cells. Our previous study showed that nanotechnology improved adipose-derived stem cell （ADSC） therapy for erectile dysfunction of cavernous nerve injury （CNI） by attracting cells in the corpus cavernosum. These results indicated the possibility of using a reduced dosage of ADSCs for intracavernous injection. In this exploratory study, we used lower dosage （2 × 105 cells） of ADSCs for intracavernous injection （ICI） and the nanotechnology approach. Intracavernous pressure and mean arterial pressure were measured at day 28 to assess erectile function. The low-dose ADSC therapy group showed favorable treatment effects, and nanotechnology further improved these effects. In vivo imaging of ICI cells revealed that the fluorescein signals of NanoShuttle-bound ADSCs （NanoADSCs） were much stronger than those of ADSCs at days 0, 1, and 3. Both immunofluorescence and Western blot analysis showed a significant increase in smooth muscle, endothelium, and nerve tissue in the ADSC group compared to that in the CNI group; further improvement was achieved with assisted nanotechnology. These findings demonstrate that nanotechnology can be used to further improve the effect of small dosage of ADSCs to improve erectile function. Abundant NanoADSCs remain in the corpus cavernosum in vivo for at least 3 days. The mechanism of erectile function improvement may be related to the regeneration of the smooth muscle, endothelium, and nerve tissues.

Ambient ozone pollution is associated with decreased semen quality:longitudinal analysis of 8945 semen samples from 2015 to 2018 and during pollution-control period in Beijing,China

Previous studies suggest that air pollution has a negative effect on semen quality.However,most studies are cross-sectional and the results are controversial.This study investigated the associations between air pollutants(PM25,PM10,SO2,NO2,CO,and O3)and seme n quality among sperm donation candidates,especially when the air pollution was artificially con trolled in Beijing,China.We an a lyzed 8945 semen samples in the human sperm bank of Peki ng Un iversity Third Hospital(Beijing,China)from October 2015 to May 2018.Air pollution data during the entire period(0-90 days prior)and key stages(0-9,10-14,and 70-90 days prior)of sperm development were collected from the China National Environmental Monitoring Centre.The association between air pollutants and seme n parameters(sperm con centrati on and progressive motility)was an a lyzed by a mixed model adjusted for age,abstinence duration,month,and average ambient temperature.Only O3 during key stages of 0-9 days and 10-14 days and the entire period was negatively associated with sperm concentration between 2015 and 2018(P v 0.01).During the period of air pollution control from November 2017 to January 2018,except for the in crease in O3 concentration,other five pollutants'concentrations decreased compared to those in previous years.In this period,the sperm concentration decreased(P<0.001).During the pollution-control period,O3 exposure 10-14 days prior was negatively associated with sperm concentration(95%Cl:-0.399—0.111;P<0.001).No significant association was found between the other five pollutants and semen quality during that period?Our study suggested that only O3 exposure was harmful to semen quality.Therefore,O3 should not be neglected during pollution control operation.

DEtail-seq is an ultra-efficient and convenient method for meiotic DNA break profiling in multiple organisms

Programmed DNA double-strand break(DSB)formation is a crucial step in meiotic recombination,yet techniques for highefficiency and precise mapping of the 3’ends of DSBs are still in their infancy.Here,we report a novel technique,named DNA End tailing and sequencing(DEtail-seq),which can directly and ultra-efficiently characterize the 3’ends of meiotic DSBs with near single-nucleotide resolution in a variety of species,including yeast,mouse,and human.We find that the 3’ends of meiotic DSBs are stable without significant resection in budding yeast.Meiotic DSBs are strongly enriched in de novo H3K4me3 peaks in the mouse genome at leptotene stage.We also profile meiotic DSBs in human and find DSB hotspots are enriched near the common fragile sites during human meiosis,especially at CCCTC-binding factor(CTCF)-associated enhancers.Therefore,DEtail-seq provides a powerful method to detect DSB ends in various species,and our results provide new insights into the distribution and regulation of meiotic DSB hotspots.

Sirtl regulates testosterone biosynthesis in Leydig cells via modulating autophagy

Dear Editor,Steroid hormones are crucial signal molecules that regulate a large number of physiological and developmental pro-cesses.Testosterone is the key steroid hormone required for the development of male characteristics and also supports the physiology of the male reproductive system(Sinclair et al.,2015).Testosterone is primarily produced by the Leydig cells residing in the testicular interstitium.The cholesterol acts as a substrate for the biosynthesis of testosterone.Since steroidogenic cells are capable of stor-ing only very ltte hormone,rapid synthesis of hormone requires the mobilization of the precursor cholesterol,chiefly stored as intracellular lipid droplets(LDs)(Danielsen et al.,2016).Leydig cells are the major sites to produce testos-terone,there are extremely active autophagy in them,and a decline in steroidogenesis has also been associated with the decline of autophagic flow.Moreover,the disruption of autophagy leads to decreased intracellular LDs,and there-fore affects testosterone synthesis in the Leydig cells(Danielsen et al..2016;Gao et al,2018).