Necrozoospermia is a poorly documented condition with a low incidence,and its definition and clinical significance are unclear.Herein,we provide a reference range for necrozoospermia and discuss its possible etiology ...Necrozoospermia is a poorly documented condition with a low incidence,and its definition and clinical significance are unclear.Herein,we provide a reference range for necrozoospermia and discuss its possible etiology and impact on male fertility and assisted reproductive outcomes.We extracted relevant information from 650 Chinese male partners of infertile couples and statistically analyzed sperm vitality.Necrozoospermia was present in 3.4%(22/650)of our study population,and the lower cut-off value for sperm vitality was 75.3%.We compared two methods for assessing sperm vitality(eosin-nigrosin head staining and hypo-osmotic swelling test[HOST]),for which the percentage in the eosin-nigrosin group(mean±standard deviation[s.d.]:77.5%±10.5%)was significantly higher than that in the HOST group(mean±s.d.:58.1%±6.7%[5-10 min after incubation]and 55.6%±8.2%[25-30 min after incubation];both P<0.001).The incidence of necrozoospermia increased with age(odds ratio[OR]=1.116,95%confidence interval[CI]:1.048-1.189,P=0.001),while the percentage of normal sperm morphology and DNA fragmentation index(DFI)were significantly associated with necrozoospermia,with ORs of 0.691(95%CI:0.511-0.935,P=0.017)and 1.281(95%CI:1.180-1.390,P<0.001),respectively.In the following 6 months,we recruited 166 patients in the nonnecrozoospermia group and 87 patients in the necrozoospermia group to compare intracytoplasmic sperm injection(ICSI)and pregnancy outcomes between the two groups.The necrozoospermia group had a significantly lower normal fertilization rate(74.7%vs 78.2%,P=0.041;OR=0.822;95%CI:0.682-0.992)than that in the nonnecrozoospermia group.This study presents substantial information on necrozoospermia to establish comprehensive and applicable reference values for sperm vitality for spontaneous conception and artificially assisted reproductive management.展开更多
The clinical applications of acrosin activity are limited.We analyzed 61578 male partners in infertile couples who visited the outpatient department of the Reproductive and Genetic Hospital of CITIC-Xiangya(Changsha,C...The clinical applications of acrosin activity are limited.We analyzed 61578 male partners in infertile couples who visited the outpatient department of the Reproductive and Genetic Hospital of CITIC-Xiangya(Changsha,China)between August 2014 and December 2019 to determine the reference ranges and thresholds for acrosin activity in infertile Chinese men;to determine whether correlations exist between acrosin activity and age,sperm concentration,sperm morphology,or sperm motility;and to evaluate whether acrosin activity could serve as an effective prognostic indicator for choosing between in vitro fertilization(IvF)and intracytoplasmic sperm injection(icsl)in the clinic.The cut-off value for the normal reference range of acrosin activity for male partners in infertile couples was 24.78μlU per 106 sperm.There was no significant association between acrosin activity and age,sperm concentration,semen volume,total sperm count,progressive motility,or total motile spermatozoa.A weak positive correlation was found between acrosin activity and normal sperm morphology.There was a statistically significant difference in abnormal acrosome morphology between the group with high acrosin activity(>24.78μlU per 106 sperm)and the group with low acrosin activity(<24.78μlU per 106 sperm).The group with a low IVF fertilization rate had a high index of abnormal acrosomal morphology at 21.2%,while the group with a high IVF fertilization rate had a low index of 0.2%.At an acrosin activity of<24.78μlU per 10 sperm,in one cycle of the same patient,the fertilization rate,normal fertilization rate,and good-quality embryo rate for Icsl were significantly higher than those for IVF.Therefore,the most promising application of acrosin activity could be in the selection of IcsI over IVF for infertile male patients with complete fertilization failure or a low fertilization rate.展开更多
Cryopreservation is a fundamental technology in biomedical research,regenerative medicine,and tissue engineering,enabling the long-term storage of cells,tissues,and organs.However,its effectiveness is limited by chall...Cryopreservation is a fundamental technology in biomedical research,regenerative medicine,and tissue engineering,enabling the long-term storage of cells,tissues,and organs.However,its effectiveness is limited by challenges such as intracellular ice formation,cryoprotectant toxicity,and reduced post-thaw viability.This review explores the crucial role of encapsulation in enhancing cryopreservation efficiency,with a focus on recent advances in materials science,bioengineering,and cryobiology.Emerging technologies,such as nanotechnology and stimuli-responsive polymers,are transforming encapsulation strategies.Innovations such as microfluidic systems offer precise control over cooling rates and cryoprotectant distribution,thereby mitigating conventional limitations.The review also addresses current obstacles related to scaling up encapsulation processes and ensuring the long-term biocompatibility and stability of preserved specimens.By synthesizing recent findings,this work provides a comprehensive resource for researchers and clinicians seeking to enhance biopreservation techniques and their applications in contemporary medicine and biotechnology.Finally,the review identifies critical knowledge gaps that must be addressed to improve the efficacy of cryopreservation strategies and advance their clinical translation.展开更多
Multiple morphological abnormalities of sperm flagella(MMAF)is a severe form of asthenoteratozoospermia,characterized by morphological abnormalities and reduced motility of sperm,causing male infertility.Although appr...Multiple morphological abnormalities of sperm flagella(MMAF)is a severe form of asthenoteratozoospermia,characterized by morphological abnormalities and reduced motility of sperm,causing male infertility.Although approximately 60%of MMAF cases can be explained genetically,the etiology of the remaining cases is unclear.Here,we identified two novel compound heterozygous variants in the gene,dynein axonemal heavy chain 10(DNAH10),in three patients from two unrelated Pakistani families using whole-exome sequencing(WES),including one compound heterozygous mutation(DNAH10:c.9409C>A[p.P3137T];c.12946G>C[p.D4316H])in family 1 and another compound heterozygous mutation(DNAH10:c.8849G>A[p.G2950D];c.11509C>T[p.R3687W])in family 2.All the identified variants are absent or rare in public genome databases and are predicted to have deleterious effects according to multiple bioinformatic tools.Sanger sequencing revealed that these variants follow an autosomal recessive mode of inheritance.Hematoxylin and eosin(H&E)staining revealed MMAF,including sperm head abnormalities,in the patients.In addition,immunofluorescence staining revealed loss of DNAH10 protein signals along sperm flagella.These findings broaden the spectrum of DNAH10 variants and expand understanding of the genetic basis of male infertility associated with the MMAF phenotype.展开更多
A previous study showed that the length of the foreskin plays a role in the risk of sexually transmitted infections and chronic prostatitis,which can lead to poor quality of sexual life.Here,the association between fo...A previous study showed that the length of the foreskin plays a role in the risk of sexually transmitted infections and chronic prostatitis,which can lead to poor quality of sexual life.Here,the association between foreskin length and sexual dysfunction was evaluated.A total of 5700 participants were recruited from the andrology clinic at The First Affiliated Hospital of University of Science and Technology of China(Hefei,China).Clinical characteristics,including foreskin length,were collected,and sexual function was assessed by the International Index of Erectile Function-5(IIEF-5)and Premature Ejaculation Diagnostic Tool(PEDT)questionnaires.Men with sexual dysfunction were more likely to have redundant foreskin than men without sexual dysfunction.Among the 2721 erectile dysfunction(ED)patients and 1064 premature ejaculation(PE)patients,301(11.1%)ED patients and 135(12.7%)PE patients had redundant foreskin,respectively.Men in the PE group were more likely to have redundant foreskin than men in the non-PE group(P=0.004).Logistic regression analyses revealed that the presence of redundant foreskin was associated with increased odds of moderate/severe ED(adjusted odds ratio[aOR]=1.31,adjusted P=0.04),moderate PE(aOR=1.38,adjusted P=0.02),and probable PE(aOR=1.37,adjusted P=0.03)after adjusting for confounding variables.Our study revealed a positive correlation between the presence of redundant foreskin and the risk of sexual dysfunction,especially in PE patients.Assessment of the length of the foreskin during routine clinical diagnosis may provide information for patients with sexual dysfunction.展开更多
Cryopreservation of rare testicular-retrieved spermatozoa for intracytoplasmic sperm injection(ICSI)in patients with severe oligozoospermia and azoospermia remains a major challenge in clinical practice.This study eva...Cryopreservation of rare testicular-retrieved spermatozoa for intracytoplasmic sperm injection(ICSI)in patients with severe oligozoospermia and azoospermia remains a major challenge in clinical practice.This study evaluated the Cryopiece system as a potential technique to cryopreserve rare human spermatozoa for ICSI.Small numbers of ejaculated(24 patients)and testicular(13 patients)spermatozoa were cryopreserved using the Cryopiece system.The total number of recovered spermatozoa and motility were assessed after thawing.Thirty-seven couples underwent ICSI using spermatozoa cryopreserved by the Cryopiece system,and ICSI outcomes(rates of fertilization,embryo cleavage,and clinical pregnancy)were evaluated.The average sperm post-thaw retrieval rate was 79.1%,and motility was 29.7%.Ejaculated spermatozoa had a higher post-thaw motility(32.5%)than testicular spermatozoa(21.8%;P=0.005).ICSI achieved a fertilization rate of 61.9%,embryo cleavage rate of 84.6%,and clinical pregnancy rate of 43.3%.The ICSI outcomes in the ejaculated and testicular frozen-thawed spermatozoa were similar.Assisted oocyte activation(AOA)after ICSI with motile(72.1%)or immotile(71.9%)spermatozoa resulted in a significantly higher fertilization rate than that when using motile spermatozoa without AOA(52.0%;P=0.005).However,AOA did not enhance the clinical pregnancy rate(55.6%or 40.0%vs 35.3%;P=0.703).The Cryopiece system is simple and useful for the cryopreservation of small numbers of ejaculated or testicular spermatozoa for ICSI in patients with severe oligozoospermia or nonobstructive azoospermia.展开更多
Objective:Hepatolenticular degeneration (HLD) is an autosomal recessive disorder that manifests as multiorgan damage due to impaired copper (Cu) metabolism. Female patients with HLD often experience reproductive impai...Objective:Hepatolenticular degeneration (HLD) is an autosomal recessive disorder that manifests as multiorgan damage due to impaired copper (Cu) metabolism. Female patients with HLD often experience reproductive impairments. This study investigated the protective effect of berberine against ovarian damage in toxic-milk (TX) mice, a murine model for HLD.Methods:Mice were categorized into control group, HLD TX group (HLD group), penicillamine (Cu chelator)-treated TX group and berberine-treated TX group. Body weight, ovary weight and the number of ovulated eggs were recorded. Follicular morphology and cellular ultrastructure were examined. Total iron, ferrous iron (Fe2+) and trivalent iron (Fe3+) levels, as well as malondialdehyde (MDA), glutathione(GSH) and oxidized glutathione (GSSG), were measured in the ovaries. Western blot analysis was used to analyze the expression of proteins related to ferroptosis and endoplasmic reticulum (ER) stress.Results:Ovarian tissue damage was evident in the HLD group, with a significant increase in ferroptosis and ER stress compared to the control group. This damage was inhibited by treatment with penicillamine,a Cu chelator. Compared with the HLD group, berberine increased the number of ovulations, and improved ovarian morphology and ultrastructure. Further, we found that berberine reduced total iron,Fe2+, MDA and GSSG levels, elevated GSH levels, decreased the expression of the ferroptosis marker protein prostaglandin-endoperoxide synthase 2 (PTGS2), and increased glutathione peroxidase 4 (GPX4)expression. Furthermore, berberine inhibited the expression of ER stress-associated proteins mediated by the protein kinase RNA-like ER kinase (PERK) pathway.Conclusion:Ferroptosis and ER stress are involved in Cu-induced ovarian damage in TX mice. Berberine ameliorates ovarian damage in HLD TX mice by inhibiting ferroptosis and ER stress.展开更多
Phospholipase C zeta(PLC)is a key sperm-borne oocyte-activating factor that triggers Ca^(2+)oscillations and the subsequent block to polyspermy following gamete fusion.Mutations in PLCZ1,the gene encoding PLCζ,cause ...Phospholipase C zeta(PLC)is a key sperm-borne oocyte-activating factor that triggers Ca^(2+)oscillations and the subsequent block to polyspermy following gamete fusion.Mutations in PLCZ1,the gene encoding PLCζ,cause male infertility and intracytoplasmic sperm injection(ICSI)fertilization failure;and PLCζ expression and localization patterns are significantly correlated with ICSI fertilization rate(FR).However,in conventional in vitro fertilization(cIVF),whether and how sperm PLCζ affects fertilization remain unclear.Herein,we identified one previously reported and two novel PLCZ1 mutations associated with polyspermy in vitro that are characterized by excessive sperm-zona binding and a delay in pronuclei(PN)formation.Immunofluorescence staining and oocyte activation testing revealed that virtually all spermatozoa from patients lacked functional PLCζ and were thus unable to evoke Ca^(2+) oscillations.ICSI with an artificial oocyte activation treatment successfully rescued the polyspermic phenotype and resulted in a live birth.Furthermore,we analyzed PLCζ in an additional 58 males after cIVF treatment in the Reproductive and Genetic Hospital of CiTiC-Xiangya(Changsha,China)between February 2019 and January 2022.We found that the proportion of spermatozoa that expressed PLCζ was positively correlated with both 2PN rate and total FR.The optimal cutoff value below which males were likely to experience low FR(total FR≤30%)after clVF was 56.7%for the proportion of spermatozoa expressing PLC5.Our study expands the mutation and the phenotypic spectrum of PLCZ1 and further suggests that PLCζ constitutes a promising biomarker for identifying low FRs cases in cIVF due to sperm-related oocyte activation deficiency and that sperm PLCζ analysis may benefit the widermale population and not onlymen with IcsI failure.展开更多
Male infertility can result from impaired sperm motility caused by multiple morphological abnormalities of the flagella(MMAF).Distinct projections encircling the central microtubules of the spermatozoal axoneme play p...Male infertility can result from impaired sperm motility caused by multiple morphological abnormalities of the flagella(MMAF).Distinct projections encircling the central microtubules of the spermatozoal axoneme play pivotal roles in flagellar bending and spermatozoal movement.Mammalian sperm-associated antigen 17(SPAG17)encodes a conserved axonemal protein of cilia and flagella,forming part of the C1a projection of the central apparatus,with functions related to ciliary/flagellar motility,skeletal growth,and male fertility.This study investigated two novel homozygous SPAG17 mutations(M1:NM_206996.2,c.829+1G>T,p.Asp212_Glu276del;and M2:c.2120del,p.Leu707*)identified in four infertile patients from two consanguineous Pakistani families.These patients displayed the MMAF phenotype confirmed by Papanicolaou staining and scanning electron microscopy assays of spermatozoa.Quantitative real-time polymerase chain reaction(PCR)of patients’spermatozoa also revealed a significant decrease in SPAG17 mRNA expression,and immunofluorescence staining showed the absence of SPAG17 protein signals along the flagella.However,no apparent ciliary-related symptoms or skeletal malformations were observed in the chest X-rays of any of the patients.Transmission electron microscopy of axoneme cross-sections from the patients showed incomplete C1a projection and a higher frequency of missing microtubule doublets 1 and 9 compared with those from fertile controls.Immunofluorescence staining and Western blot analyses of spermatogenesis-associated protein 17(SPATA17),a component of the C1a projection,and sperm-associated antigen 6(SPAG6),a marker of the spring layer,revealed disrupted expression of both proteins in the patients’spermatozoa.Altogether,these findings demonstrated that SPAG17 maintains the integrity of spermatozoal flagellar axoneme,expanding the phenotypic spectrum of SPAG17 mutations in humans.展开更多
Recent studies have shown that shorter periods of ejaculatory abstinence may enhance certain sperm parameters,but the molecular mechanisms underlying these improvements are still unclear.This study explored whether re...Recent studies have shown that shorter periods of ejaculatory abstinence may enhance certain sperm parameters,but the molecular mechanisms underlying these improvements are still unclear.This study explored whether reduced abstinence periods could improve semen quality,particularly for use in assisted reproductive technologies(ART).We analyzed semen samples from men with normal sperm counts(n=101)and those with low sperm motility or concentration(n=53)after 3-7 days of abstinence and then after 1-3h of abstinence,obtained from the Reproductive&Genetic Hospital of CITIC-Xiangya(Changsha,China).Physiological and biochemical sperm parameters were evaluated,and the dynamics of transfer RNA(tRNA)-derived fragments(tRFs)were analyzed using deep RNA sequencing in five consecutive samples from men with normal sperm counts.Our results revealed significant improvement in sperm motility and a decrease in the DNA fragmentation index after the 1-to 3-h abstinence period.Additionally,we identified 245 differentially expressed tRFs,and the mitogen-activated protein kinase(MAPK)signaling pathway was the most enriched.Further investigations showed significant changes in tRF-Lys-TTT and its target gene mitogen-activated protein kinase kinase 2(MAP2K2),which indicates a role of tRFs in improving sperm function.These findings provide new insights into how shorter abstinence periods influence sperm quality and suggest that tRFs may serve as biomarkers for male fertility.This research highlights the potential for optimizingART protocols and improving reproductive outcomes through molecular approaches that target sperm function.展开更多
Oocyte cryopreservation is an essential procedure in assisted reproductive technologies,aimed at preserving fertility,particularly for women undergoing IVF treatment or at risk of ovarian damage due to radiation,chemo...Oocyte cryopreservation is an essential procedure in assisted reproductive technologies,aimed at preserving fertility,particularly for women undergoing IVF treatment or at risk of ovarian damage due to radiation,chemotherapy,or surgery.Despite its growing use,the survival and fertilization rates of cryopreserved oocytes remain suboptimal,largely due to cryo-induced oxidative stress.The generation of Reactive Oxygen Species(ROS)during freezing and thawing causes considerable damage to key cellular components,including proteins,lipids,DNA,and mitochondria.This oxidative stress compromises oocyte quality and reduces developmental potential.To address these challenges,the use of additives-especially antioxidants-has shown significant promise in mitigating oxidative damage.Enzymatic antioxidants such as Superoxide Dismutase(SOD)and Catalase(CAT),along with non-enzymatic antioxidants like glutathione,melatonin,and resveratrol,have demonstrated the ability to neutralize ROS and improve oocyte viability and developmental outcomes.Recent studies highlight the potential of Mitoquinone(MitoQ),a mitochondria-targeted antioxidant,to effectively counteract mitochondrial ROS and enhance cellular defense mechanisms during cryopreservation.This review explores the cellular mechanisms of cryodamage,the role of oxidative stress in oocyte cryopreservation,and the potential of various antioxidant strategies to enhance oocyte survival and function.Developing effective antioxidant supplementation approaches may significantly improve the outcomes of cryopreservation in reproductive medicine.展开更多
Multiple morphological abnormalities of the flagella(MMAF)represent a severe form of sperm defects leading to asthenozoospermia and male infertility.In this study,we identified a novel homozygous splicing mutation(c.8...Multiple morphological abnormalities of the flagella(MMAF)represent a severe form of sperm defects leading to asthenozoospermia and male infertility.In this study,we identified a novel homozygous splicing mutation(c.871-4 ACA>A)in the adenylate kinase 7(AK7)gene by whole-exome sequencing in infertile individuals.Spermatozoa from affected individuals exhibited typical MMAF characteristics,including coiled,bent,short,absent,and irregular flagella.Transmission electron microscopy analysis showed disorganized axonemal structure and abnormal mitochondrial sheets in sperm flagella.Immunofluorescence staining confirmed the absence of AK7 protein from the patients’spermatozoa,validating the pathogenic nature of the mutation.This study provides direct evidence linking the AK7 gene to MMAF-associated asthenozoospermia in humans,expanding the mutational spectrum of AK7 and enhancing our understanding of the genetic basis of male infertility.展开更多
Spermiogenesis is a complex and tightly regulated process,consisting of acrosomal biogenesis,condensation of chromatin,flagellar assembly,and disposal of extra cytoplasm.Previous studies have reported that sperm flage...Spermiogenesis is a complex and tightly regulated process,consisting of acrosomal biogenesis,condensation of chromatin,flagellar assembly,and disposal of extra cytoplasm.Previous studies have reported that sperm flagellar 2(SPEF2)deficiency causes severe asthenoteratozoospermia owing to spermiogenesis failure,but the underlying molecular mechanism in humans remains unclear.Here,we performed proteomic analysis on spermatozoa from three SPEF2 mutant patients to study the functional role of SPEF2 during sperm tail development.A total of 1262 differentially expressed proteins were detected,including 486 upregulated and 776 downregulated.The constructed heat map of the differentially expressed proteins showed similar trends.Among these,the expression of proteins related to flagellar assembly,including SPEF2,sperm associated antigen 6(SPAG6),dynein light chain tctex-type 1(DYNLT1),radial spoke head component 1(RSPH1),translocase of outer mitochondrial membrane 20(TOM20),EF-hand domain containing 1(EFHC1),meiosis-specific nuclear structural 1(MNS1)and intraflagellar transport 20(IFT20),was verified by western blot.Functional clustering analysis indicated that these differentially expressed proteins were specifically enriched for terms such as spermatid development and flagellar assembly.Furthermore,we showed that SPEF2 interacts with radial spoke head component 9(RSPH9)and IFT20 in vitro,which are well-studied components of radial spokes or intra-flagellar transport and are essential for flagellar assembly.These results provide a rich resource for further investigation into the molecular mechanism underlying the role that SPEF2 plays in sperm tail development and could provide a theoretical basis for gene therapy in SPEF2 mutant patients in the future.展开更多
Sperm morphology was once believed as one of the most predictive indicators of pregnancy outcome in assisted reproductive technology(ART).However,the impact of teratozoospermia on in vitro fertilization(IVF)/intracyto...Sperm morphology was once believed as one of the most predictive indicators of pregnancy outcome in assisted reproductive technology(ART).However,the impact of teratozoospermia on in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI)outcomes and its offspring remains inconclusive.In order to evaluate the influence of teratozoospermia on pregnancy outcome and newborn status after IVF and ICSI,a retrospective study was conducted.This was a matched case-control study that included 2202 IVF cycles and 2574 ICSI cycles and was conducted at the Reproductive and Genetic Hospital of CITIC-Xiangya in Changsha,China,from June 2013 to June 2018.Patients were divided into two groups based on sperm morphology:teratozoospermia and normal sperm group.The pregnancy outcome and newborn outcome were analyzed.The results indicated that couples with teratozoospermia had a significantly lower optimal embryo rate compared to those with normal sperm morphology in IVF(P=0.007),while there were no statistically significant differences between the two groups in terms of the fertilization rate,cleavage rate,implantation rate,and pregnancy rate(all P>0.05).Additionally,teratozoospermia was associated with lower infant birth weight in multiple births after IVF.With regard to ICSI,there was no significant difference in both pregnancy outcome and newborn outcome between the teratozoospermia and normal groups(both P>0.05).Furthermore,no increase in the risk of birth defects occurred in the teratozoospermia group after IVF/ICSI.Consequently,we believe that teratozoospermia has limited predictive value for pregnancy outcomes in IVF/ICSI,and has little impact on the resulting offspring if multiple pregnancy is avoided.展开更多
Diminished ovarian reserve(DOR)is a disorder of ovarian function in which the ovary loses its normal reproductive potential,including decreasing oocyte quantity and quality.The disorder is associated with infertility ...Diminished ovarian reserve(DOR)is a disorder of ovarian function in which the ovary loses its normal reproductive potential,including decreasing oocyte quantity and quality.The disorder is associated with infertility in women of reproductive age with regular menses and poor response to controlled ovarian hyperstimulation(Broekmans et al.,2009).展开更多
BACKGROUND Spermatogonial stem cells(SSCs)are the origin of male spermatogenesis,which can reconstruct germ cell lineage in mice.However,the application of SSCs for male fertility restoration is hindered due to the un...BACKGROUND Spermatogonial stem cells(SSCs)are the origin of male spermatogenesis,which can reconstruct germ cell lineage in mice.However,the application of SSCs for male fertility restoration is hindered due to the unclear mechanisms of proliferation and self-renewal in humans.AIM To investigate the role and mechanism of SPOC domain-containing protein 1(SPOCD1)in human SSC proliferation.METHODS We analyzed publicly available human testis single-cell RNA sequencing(RNAseq)data and found that SPOCD1 is predominantly expressed in SSCs in the early developmental stages.Small interfering RNA was applied to suppress SPOCD1 expression to detect the impacts of SPOCD1 inhibition on SSC proliferation and apoptosis.Subsequently,we explored the target genes of SPOCD1 using RNA-seq and confirmed their role by restoring the expression of the target genes.In addition,we examined SPOCD1 expression in some non-obstructive azoospermia(NOA)patients to explore the correlation between SPOCD1 and NOA.RESULTS The uniform manifold approximation and projection clustering and pseudotime analysis showed that SPOCD1 was highly expressed in the early stages of SSC,and immunohistological results showed that SPOCD1 was mainly localized in glial cell line-derived neurotrophic factor family receptor alpha-1 positive SSCs.SPOCD1 knockdown significantly inhibited cell proliferation and promoted apoptosis.RNA-seq results showed that SPOCD1 knockdown significantly downregulated genes such as adenylate kinase 4(AK4).Overexpression of AK4 in SPOCD1 knockdown cells partially reversed the phenotypic changes,indicating that AK4 is a functional target gene of SPOCD1.In addition,we found a significant downregulation of SPOCD1 expression in some NOA patients,suggesting that the downregulation of SPOCD1 may be relevant for NOA.CONCLUSION Our study broadens the understanding of human SSC fate determination and may offer new theories on the etiology of male infertility.展开更多
Congenital bilateral absence of the vas deferens(CBAVD)is observed in 1%–2%of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator(CFTR)mutations.CFTR i...Congenital bilateral absence of the vas deferens(CBAVD)is observed in 1%–2%of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator(CFTR)mutations.CFTR is one of the most well-known genes related to male fertility.The frequency of CFTR mutations or impaired CFTR expression is increased in men with nonobstructive azoospermia(NOA).CFTR mutations are highly polymorphic and have established ethnic specificity.Compared with F508Del in Caucasians,the p.G970D mutation is reported to be the most frequent CFTR mutation in Chinese patients with cystic fibrosis.However,whether p.G970D participates in male infertility remains unknown.Herein,a loss-of-function CFTR p.G970D missense mutation was identified in a patient with CBAVD and NOA.Subsequent retrospective analysis of 122 Chinese patients with CBAVD showed that the mutation is a common pathogenic mutation(4.1%,5/122),excluding polymorphic sites.Furthermore,we generated model cell lines derived from mouse testes harboring the homozygous Cftr p.G965D mutation equivalent to the CFTR variant in patients.The Cftr p.G965D mutation may be lethal in spermatogonial stem cells and spermatogonia and affect the proliferation of spermatocytes and Sertoli cells.In spermatocyte GC-2(spd)ts(GC2)Cftr p.G965D cells,RNA splicing variants were detected and CFTR expression decreased,which may contribute to the phenotypes associated with impaired spermatogenesis.Thus,this study indicated that the CFTR p.G970D missense mutation might be a pathogenic mutation for CBAVD in Chinese males and associated with impaired spermatogenesis by affecting the proliferation of germ cells.展开更多
Dear Editor, There are two main causes of severe asthenozoospermia： ultrastructural defects （genetically inherited and congenital defects） of the sperm flagellum and necrozoospermia or sperm degeneration secondary ...Dear Editor, There are two main causes of severe asthenozoospermia： ultrastructural defects （genetically inherited and congenital defects） of the sperm flagellum and necrozoospermia or sperm degeneration secondary to other pathological changes （see review by Ortega et al2）. Genetic-related absolute asthenozoospermia includes primary ciliary dyskinesia and dysplasia of the fibrous sheath （DFS）, which result in 100% （or nearly） immotile spermatozoa.展开更多
Male infertility is a major reproductive disorder,which is clinically characterized by highly heterogeneous phenotypes of abnormal sperm count or quality.To date,five male patients with biallelic loss-of-function(LOF)...Male infertility is a major reproductive disorder,which is clinically characterized by highly heterogeneous phenotypes of abnormal sperm count or quality.To date,five male patients with biallelic loss-of-function(LOF)variants of PARN-like ribonuclease domain-containing exonuclease 1(PNLDC1)have been reported to experience infertility with nonobstructive azoospermia.The aim of this study was to identify the genetic cause of male infertility with oligo-astheno-teratozoospermia(OAT)in a patient from a Chinese Han family.Whole-exome and Sanger sequencing analyses identified a homozygous LOF variant(NM_173516.2,c.l42C>T,p.Gln48Ter)in PNLDC1.Hematoxylin and eosin staining revealed that the spermatozoa of the patient with OAT had an irregular head phenotype,including microcephaly,head tapering,and globozoospermia.Consistently,peanut agglutinin staining of the spermatozoa revealed a complete or partial loss of the acrosome.Furthermore,the disomy rate of chromosomes in the patient’s spermatozoa was significantly increased compared with that of a fertile control sample.We reported an LOF variant of the PNLDC1 gene responsible for OAT.展开更多
Pericentric inversion of chromosome 9(inv[9])is a common chromosomal structural variant,but its impact on clinical outcomesremains debated.The screening criteria of sperm banks are rarely mentioned to individuals with...Pericentric inversion of chromosome 9(inv[9])is a common chromosomal structural variant,but its impact on clinical outcomesremains debated.The screening criteria of sperm banks are rarely mentioned to individuals with inv(9).In this study,we evaluatedthe fertility of sperm donors with inv(9)who met eligibility criteria for sperm banks(inv[9]-eligible donors).From March 2004 toMay 2022,chromosomal analysis of 16124 sperm donors at CITIC-Xiangya Human Sperm Bank in Hunan Province(Changsha,China)found that 251(1.6%)had chromosome variations,with inv(9)being the most prevalent at 1.1%.All 169 inv(9)-eligibledonors were contacted to collect fertility outcome data,along with 206 eligible donors without inv(9)as controls.In addition,semen samples from inv(9)-eligible donors and eligible donors underwent assessments of sperm fluorescence in situ hybridization(FISH),mitochondrial membrane potential,DNA fragmentation index,acrosome integrity,reactive oxygen species(ROS),andsperm morphology.Results showed that inv(9)did not significantly increase reproductive risks overall.Despite detecting ROSlevel differences,the clinical impact may be insignificant.This study provides new data on the inv(9)population that can serveas a valuable reference for decision-making by sperm banks as well as for genetic counseling and clinical guidance for individualscarrying inv(9)variant.展开更多
基金supported by the Hunan Province Municipal Natural Science Foundation(No.2022JJ30018 to WNL)the Hunan Province Health Commission Science Foundation(No.B202301037899 to WNL)Research Grant of CITIC-Xiangya(No.YNXM-202219 to WNL).
文摘Necrozoospermia is a poorly documented condition with a low incidence,and its definition and clinical significance are unclear.Herein,we provide a reference range for necrozoospermia and discuss its possible etiology and impact on male fertility and assisted reproductive outcomes.We extracted relevant information from 650 Chinese male partners of infertile couples and statistically analyzed sperm vitality.Necrozoospermia was present in 3.4%(22/650)of our study population,and the lower cut-off value for sperm vitality was 75.3%.We compared two methods for assessing sperm vitality(eosin-nigrosin head staining and hypo-osmotic swelling test[HOST]),for which the percentage in the eosin-nigrosin group(mean±standard deviation[s.d.]:77.5%±10.5%)was significantly higher than that in the HOST group(mean±s.d.:58.1%±6.7%[5-10 min after incubation]and 55.6%±8.2%[25-30 min after incubation];both P<0.001).The incidence of necrozoospermia increased with age(odds ratio[OR]=1.116,95%confidence interval[CI]:1.048-1.189,P=0.001),while the percentage of normal sperm morphology and DNA fragmentation index(DFI)were significantly associated with necrozoospermia,with ORs of 0.691(95%CI:0.511-0.935,P=0.017)and 1.281(95%CI:1.180-1.390,P<0.001),respectively.In the following 6 months,we recruited 166 patients in the nonnecrozoospermia group and 87 patients in the necrozoospermia group to compare intracytoplasmic sperm injection(ICSI)and pregnancy outcomes between the two groups.The necrozoospermia group had a significantly lower normal fertilization rate(74.7%vs 78.2%,P=0.041;OR=0.822;95%CI:0.682-0.992)than that in the nonnecrozoospermia group.This study presents substantial information on necrozoospermia to establish comprehensive and applicable reference values for sperm vitality for spontaneous conception and artificially assisted reproductive management.
基金This study was supported by the Hunan Province Municipal Natural Science Foundation(2022JJ30018)to WNLHunan Province Health Commission Science Foundation(B202301037899)to WNLResearch Grant of CITIC-Xiangya(YNXM-201904)to WNL.
文摘The clinical applications of acrosin activity are limited.We analyzed 61578 male partners in infertile couples who visited the outpatient department of the Reproductive and Genetic Hospital of CITIC-Xiangya(Changsha,China)between August 2014 and December 2019 to determine the reference ranges and thresholds for acrosin activity in infertile Chinese men;to determine whether correlations exist between acrosin activity and age,sperm concentration,sperm morphology,or sperm motility;and to evaluate whether acrosin activity could serve as an effective prognostic indicator for choosing between in vitro fertilization(IvF)and intracytoplasmic sperm injection(icsl)in the clinic.The cut-off value for the normal reference range of acrosin activity for male partners in infertile couples was 24.78μlU per 106 sperm.There was no significant association between acrosin activity and age,sperm concentration,semen volume,total sperm count,progressive motility,or total motile spermatozoa.A weak positive correlation was found between acrosin activity and normal sperm morphology.There was a statistically significant difference in abnormal acrosome morphology between the group with high acrosin activity(>24.78μlU per 106 sperm)and the group with low acrosin activity(<24.78μlU per 106 sperm).The group with a low IVF fertilization rate had a high index of abnormal acrosomal morphology at 21.2%,while the group with a high IVF fertilization rate had a low index of 0.2%.At an acrosin activity of<24.78μlU per 10 sperm,in one cycle of the same patient,the fertilization rate,normal fertilization rate,and good-quality embryo rate for Icsl were significantly higher than those for IVF.Therefore,the most promising application of acrosin activity could be in the selection of IcsI over IVF for infertile male patients with complete fertilization failure or a low fertilization rate.
基金supported by the National Natural Science Foundation of China(82172114)the"Challenge and Response"project for key and common technology research of Hefei(GJ2022SH08).
文摘Cryopreservation is a fundamental technology in biomedical research,regenerative medicine,and tissue engineering,enabling the long-term storage of cells,tissues,and organs.However,its effectiveness is limited by challenges such as intracellular ice formation,cryoprotectant toxicity,and reduced post-thaw viability.This review explores the crucial role of encapsulation in enhancing cryopreservation efficiency,with a focus on recent advances in materials science,bioengineering,and cryobiology.Emerging technologies,such as nanotechnology and stimuli-responsive polymers,are transforming encapsulation strategies.Innovations such as microfluidic systems offer precise control over cooling rates and cryoprotectant distribution,thereby mitigating conventional limitations.The review also addresses current obstacles related to scaling up encapsulation processes and ensuring the long-term biocompatibility and stability of preserved specimens.By synthesizing recent findings,this work provides a comprehensive resource for researchers and clinicians seeking to enhance biopreservation techniques and their applications in contemporary medicine and biotechnology.Finally,the review identifies critical knowledge gaps that must be addressed to improve the efficacy of cryopreservation strategies and advance their clinical translation.
基金supported by the National Natural Science Foundation of China(No.32100689)the National Key Research and Development Program of China(No.2021YFC2700202,No.2022YFA0806303,and No.2022YFC2702601)+1 种基金the Global Select Project(No.DJK-LX-2022010)of the Institute of Health and MedicineHefei Comprehensive National Science Center,and the Joint Fund for New Medicine of USTC(No.YD9100002034).
文摘Multiple morphological abnormalities of sperm flagella(MMAF)is a severe form of asthenoteratozoospermia,characterized by morphological abnormalities and reduced motility of sperm,causing male infertility.Although approximately 60%of MMAF cases can be explained genetically,the etiology of the remaining cases is unclear.Here,we identified two novel compound heterozygous variants in the gene,dynein axonemal heavy chain 10(DNAH10),in three patients from two unrelated Pakistani families using whole-exome sequencing(WES),including one compound heterozygous mutation(DNAH10:c.9409C>A[p.P3137T];c.12946G>C[p.D4316H])in family 1 and another compound heterozygous mutation(DNAH10:c.8849G>A[p.G2950D];c.11509C>T[p.R3687W])in family 2.All the identified variants are absent or rare in public genome databases and are predicted to have deleterious effects according to multiple bioinformatic tools.Sanger sequencing revealed that these variants follow an autosomal recessive mode of inheritance.Hematoxylin and eosin(H&E)staining revealed MMAF,including sperm head abnormalities,in the patients.In addition,immunofluorescence staining revealed loss of DNAH10 protein signals along sperm flagella.These findings broaden the spectrum of DNAH10 variants and expand understanding of the genetic basis of male infertility associated with the MMAF phenotype.
基金supported by the National Natural Science Foundation of China(No.81901543 and No.81971333)the Key Research and Development Project of Anhui Province(2022e07020014)the Joint Fund for Medical Artificial Intelligence(MAI2022Q010).
文摘A previous study showed that the length of the foreskin plays a role in the risk of sexually transmitted infections and chronic prostatitis,which can lead to poor quality of sexual life.Here,the association between foreskin length and sexual dysfunction was evaluated.A total of 5700 participants were recruited from the andrology clinic at The First Affiliated Hospital of University of Science and Technology of China(Hefei,China).Clinical characteristics,including foreskin length,were collected,and sexual function was assessed by the International Index of Erectile Function-5(IIEF-5)and Premature Ejaculation Diagnostic Tool(PEDT)questionnaires.Men with sexual dysfunction were more likely to have redundant foreskin than men without sexual dysfunction.Among the 2721 erectile dysfunction(ED)patients and 1064 premature ejaculation(PE)patients,301(11.1%)ED patients and 135(12.7%)PE patients had redundant foreskin,respectively.Men in the PE group were more likely to have redundant foreskin than men in the non-PE group(P=0.004).Logistic regression analyses revealed that the presence of redundant foreskin was associated with increased odds of moderate/severe ED(adjusted odds ratio[aOR]=1.31,adjusted P=0.04),moderate PE(aOR=1.38,adjusted P=0.02),and probable PE(aOR=1.37,adjusted P=0.03)after adjusting for confounding variables.Our study revealed a positive correlation between the presence of redundant foreskin and the risk of sexual dysfunction,especially in PE patients.Assessment of the length of the foreskin during routine clinical diagnosis may provide information for patients with sexual dysfunction.
基金supported by the National Key R&D Program of China(No.2017YFC1002003)the National Natural Science Foundation of China(No.81771637 and No.81571488)+2 种基金Shanghai Collaborative Innovation Center for Translational Medicine(No.TM202015 to ZJZ)the China Postdoctoral Science Foundation(No.2019M661521)the Key Project of Research and Development of Ningxia Hui Autonomous Region of China(No.2020BFH02002).
文摘Cryopreservation of rare testicular-retrieved spermatozoa for intracytoplasmic sperm injection(ICSI)in patients with severe oligozoospermia and azoospermia remains a major challenge in clinical practice.This study evaluated the Cryopiece system as a potential technique to cryopreserve rare human spermatozoa for ICSI.Small numbers of ejaculated(24 patients)and testicular(13 patients)spermatozoa were cryopreserved using the Cryopiece system.The total number of recovered spermatozoa and motility were assessed after thawing.Thirty-seven couples underwent ICSI using spermatozoa cryopreserved by the Cryopiece system,and ICSI outcomes(rates of fertilization,embryo cleavage,and clinical pregnancy)were evaluated.The average sperm post-thaw retrieval rate was 79.1%,and motility was 29.7%.Ejaculated spermatozoa had a higher post-thaw motility(32.5%)than testicular spermatozoa(21.8%;P=0.005).ICSI achieved a fertilization rate of 61.9%,embryo cleavage rate of 84.6%,and clinical pregnancy rate of 43.3%.The ICSI outcomes in the ejaculated and testicular frozen-thawed spermatozoa were similar.Assisted oocyte activation(AOA)after ICSI with motile(72.1%)or immotile(71.9%)spermatozoa resulted in a significantly higher fertilization rate than that when using motile spermatozoa without AOA(52.0%;P=0.005).However,AOA did not enhance the clinical pregnancy rate(55.6%or 40.0%vs 35.3%;P=0.703).The Cryopiece system is simple and useful for the cryopreservation of small numbers of ejaculated or testicular spermatozoa for ICSI in patients with severe oligozoospermia or nonobstructive azoospermia.
基金supported by the National Natural Science Foundation of China (No. 82374212, No. 81971446 and No. 81673811)the Clinical Special Translational Project of the Anhui Provincial Science and Technology Department (No. 202204295107020044)the Key Project of the Anhui Provincial Department of Education (No. 2022AH050523)。
文摘Objective:Hepatolenticular degeneration (HLD) is an autosomal recessive disorder that manifests as multiorgan damage due to impaired copper (Cu) metabolism. Female patients with HLD often experience reproductive impairments. This study investigated the protective effect of berberine against ovarian damage in toxic-milk (TX) mice, a murine model for HLD.Methods:Mice were categorized into control group, HLD TX group (HLD group), penicillamine (Cu chelator)-treated TX group and berberine-treated TX group. Body weight, ovary weight and the number of ovulated eggs were recorded. Follicular morphology and cellular ultrastructure were examined. Total iron, ferrous iron (Fe2+) and trivalent iron (Fe3+) levels, as well as malondialdehyde (MDA), glutathione(GSH) and oxidized glutathione (GSSG), were measured in the ovaries. Western blot analysis was used to analyze the expression of proteins related to ferroptosis and endoplasmic reticulum (ER) stress.Results:Ovarian tissue damage was evident in the HLD group, with a significant increase in ferroptosis and ER stress compared to the control group. This damage was inhibited by treatment with penicillamine,a Cu chelator. Compared with the HLD group, berberine increased the number of ovulations, and improved ovarian morphology and ultrastructure. Further, we found that berberine reduced total iron,Fe2+, MDA and GSSG levels, elevated GSH levels, decreased the expression of the ferroptosis marker protein prostaglandin-endoperoxide synthase 2 (PTGS2), and increased glutathione peroxidase 4 (GPX4)expression. Furthermore, berberine inhibited the expression of ER stress-associated proteins mediated by the protein kinase RNA-like ER kinase (PERK) pathway.Conclusion:Ferroptosis and ER stress are involved in Cu-induced ovarian damage in TX mice. Berberine ameliorates ovarian damage in HLD TX mice by inhibiting ferroptosis and ER stress.
基金supported by grants from the National Natural Science Foundation of China(No.81901553 to CD)the Natural Science Foundation of Hunan Province,China(No.2021JJ30461 to CD).
文摘Phospholipase C zeta(PLC)is a key sperm-borne oocyte-activating factor that triggers Ca^(2+)oscillations and the subsequent block to polyspermy following gamete fusion.Mutations in PLCZ1,the gene encoding PLCζ,cause male infertility and intracytoplasmic sperm injection(ICSI)fertilization failure;and PLCζ expression and localization patterns are significantly correlated with ICSI fertilization rate(FR).However,in conventional in vitro fertilization(cIVF),whether and how sperm PLCζ affects fertilization remain unclear.Herein,we identified one previously reported and two novel PLCZ1 mutations associated with polyspermy in vitro that are characterized by excessive sperm-zona binding and a delay in pronuclei(PN)formation.Immunofluorescence staining and oocyte activation testing revealed that virtually all spermatozoa from patients lacked functional PLCζ and were thus unable to evoke Ca^(2+) oscillations.ICSI with an artificial oocyte activation treatment successfully rescued the polyspermic phenotype and resulted in a live birth.Furthermore,we analyzed PLCζ in an additional 58 males after cIVF treatment in the Reproductive and Genetic Hospital of CiTiC-Xiangya(Changsha,China)between February 2019 and January 2022.We found that the proportion of spermatozoa that expressed PLCζ was positively correlated with both 2PN rate and total FR.The optimal cutoff value below which males were likely to experience low FR(total FR≤30%)after clVF was 56.7%for the proportion of spermatozoa expressing PLC5.Our study expands the mutation and the phenotypic spectrum of PLCZ1 and further suggests that PLCζ constitutes a promising biomarker for identifying low FRs cases in cIVF due to sperm-related oocyte activation deficiency and that sperm PLCζ analysis may benefit the widermale population and not onlymen with IcsI failure.
基金supported by the National Natural Science Foundation of China(No.82171599 and No.32270901)the National Key Research and Developmental Program of China(2022YFC2702601 and 2022YFA0806303)the Global Select Project(DJKLX-2022010)of the Institute of Health and Medicine,Hefei Comprehensive National Science Center.
文摘Male infertility can result from impaired sperm motility caused by multiple morphological abnormalities of the flagella(MMAF).Distinct projections encircling the central microtubules of the spermatozoal axoneme play pivotal roles in flagellar bending and spermatozoal movement.Mammalian sperm-associated antigen 17(SPAG17)encodes a conserved axonemal protein of cilia and flagella,forming part of the C1a projection of the central apparatus,with functions related to ciliary/flagellar motility,skeletal growth,and male fertility.This study investigated two novel homozygous SPAG17 mutations(M1:NM_206996.2,c.829+1G>T,p.Asp212_Glu276del;and M2:c.2120del,p.Leu707*)identified in four infertile patients from two consanguineous Pakistani families.These patients displayed the MMAF phenotype confirmed by Papanicolaou staining and scanning electron microscopy assays of spermatozoa.Quantitative real-time polymerase chain reaction(PCR)of patients’spermatozoa also revealed a significant decrease in SPAG17 mRNA expression,and immunofluorescence staining showed the absence of SPAG17 protein signals along the flagella.However,no apparent ciliary-related symptoms or skeletal malformations were observed in the chest X-rays of any of the patients.Transmission electron microscopy of axoneme cross-sections from the patients showed incomplete C1a projection and a higher frequency of missing microtubule doublets 1 and 9 compared with those from fertile controls.Immunofluorescence staining and Western blot analyses of spermatogenesis-associated protein 17(SPATA17),a component of the C1a projection,and sperm-associated antigen 6(SPAG6),a marker of the spring layer,revealed disrupted expression of both proteins in the patients’spermatozoa.Altogether,these findings demonstrated that SPAG17 maintains the integrity of spermatozoal flagellar axoneme,expanding the phenotypic spectrum of SPAG17 mutations in humans.
基金supported by grants from the National Key R&D Program of China(2022YFC2702700)Natural Science Foundation of Hunan Province(2024JJ6725 and 2022JJ40657)+1 种基金Hunan Provincial Grant for Innovative Province Construction(2019SK4012)the Reproductive and Genetic Hospital of CITIC-Xiangya Foundation(YNXM-202003).
文摘Recent studies have shown that shorter periods of ejaculatory abstinence may enhance certain sperm parameters,but the molecular mechanisms underlying these improvements are still unclear.This study explored whether reduced abstinence periods could improve semen quality,particularly for use in assisted reproductive technologies(ART).We analyzed semen samples from men with normal sperm counts(n=101)and those with low sperm motility or concentration(n=53)after 3-7 days of abstinence and then after 1-3h of abstinence,obtained from the Reproductive&Genetic Hospital of CITIC-Xiangya(Changsha,China).Physiological and biochemical sperm parameters were evaluated,and the dynamics of transfer RNA(tRNA)-derived fragments(tRFs)were analyzed using deep RNA sequencing in five consecutive samples from men with normal sperm counts.Our results revealed significant improvement in sperm motility and a decrease in the DNA fragmentation index after the 1-to 3-h abstinence period.Additionally,we identified 245 differentially expressed tRFs,and the mitogen-activated protein kinase(MAPK)signaling pathway was the most enriched.Further investigations showed significant changes in tRF-Lys-TTT and its target gene mitogen-activated protein kinase kinase 2(MAP2K2),which indicates a role of tRFs in improving sperm function.These findings provide new insights into how shorter abstinence periods influence sperm quality and suggest that tRFs may serve as biomarkers for male fertility.This research highlights the potential for optimizingART protocols and improving reproductive outcomes through molecular approaches that target sperm function.
基金Anhui Province Clinical Medical Research Translation Special Program(No.2204295107020002).
文摘Oocyte cryopreservation is an essential procedure in assisted reproductive technologies,aimed at preserving fertility,particularly for women undergoing IVF treatment or at risk of ovarian damage due to radiation,chemotherapy,or surgery.Despite its growing use,the survival and fertilization rates of cryopreserved oocytes remain suboptimal,largely due to cryo-induced oxidative stress.The generation of Reactive Oxygen Species(ROS)during freezing and thawing causes considerable damage to key cellular components,including proteins,lipids,DNA,and mitochondria.This oxidative stress compromises oocyte quality and reduces developmental potential.To address these challenges,the use of additives-especially antioxidants-has shown significant promise in mitigating oxidative damage.Enzymatic antioxidants such as Superoxide Dismutase(SOD)and Catalase(CAT),along with non-enzymatic antioxidants like glutathione,melatonin,and resveratrol,have demonstrated the ability to neutralize ROS and improve oocyte viability and developmental outcomes.Recent studies highlight the potential of Mitoquinone(MitoQ),a mitochondria-targeted antioxidant,to effectively counteract mitochondrial ROS and enhance cellular defense mechanisms during cryopreservation.This review explores the cellular mechanisms of cryodamage,the role of oxidative stress in oocyte cryopreservation,and the potential of various antioxidant strategies to enhance oocyte survival and function.Developing effective antioxidant supplementation approaches may significantly improve the outcomes of cryopreservation in reproductive medicine.
基金supported by the National Key Research and Development Program of China(No.2021YFC2700202,No.2022YFA0806303,and No.2022YFC2702601)the Global Select Project of the Institute of Health and Medicine,Hefei Comprehensive National Science Center(DJK-LX-2022010)the Joint Fund for New Medicine of USTC(YD9100002034).
文摘Multiple morphological abnormalities of the flagella(MMAF)represent a severe form of sperm defects leading to asthenozoospermia and male infertility.In this study,we identified a novel homozygous splicing mutation(c.871-4 ACA>A)in the adenylate kinase 7(AK7)gene by whole-exome sequencing in infertile individuals.Spermatozoa from affected individuals exhibited typical MMAF characteristics,including coiled,bent,short,absent,and irregular flagella.Transmission electron microscopy analysis showed disorganized axonemal structure and abnormal mitochondrial sheets in sperm flagella.Immunofluorescence staining confirmed the absence of AK7 protein from the patients’spermatozoa,validating the pathogenic nature of the mutation.This study provides direct evidence linking the AK7 gene to MMAF-associated asthenozoospermia in humans,expanding the mutational spectrum of AK7 and enhancing our understanding of the genetic basis of male infertility.
基金support of the National Key Research&Developmental Program of China(2018YFC1004900 to YQT)the National Natural Science Foundation of China(81971447 to YQT)the Key Grant of Prevention and Treatment of Birth Defect from Hunan province(2019SK1012 to YQT),and the research grant ofCITIC-Xiangya(YNXM-202004,YNXM-202006).
文摘Spermiogenesis is a complex and tightly regulated process,consisting of acrosomal biogenesis,condensation of chromatin,flagellar assembly,and disposal of extra cytoplasm.Previous studies have reported that sperm flagellar 2(SPEF2)deficiency causes severe asthenoteratozoospermia owing to spermiogenesis failure,but the underlying molecular mechanism in humans remains unclear.Here,we performed proteomic analysis on spermatozoa from three SPEF2 mutant patients to study the functional role of SPEF2 during sperm tail development.A total of 1262 differentially expressed proteins were detected,including 486 upregulated and 776 downregulated.The constructed heat map of the differentially expressed proteins showed similar trends.Among these,the expression of proteins related to flagellar assembly,including SPEF2,sperm associated antigen 6(SPAG6),dynein light chain tctex-type 1(DYNLT1),radial spoke head component 1(RSPH1),translocase of outer mitochondrial membrane 20(TOM20),EF-hand domain containing 1(EFHC1),meiosis-specific nuclear structural 1(MNS1)and intraflagellar transport 20(IFT20),was verified by western blot.Functional clustering analysis indicated that these differentially expressed proteins were specifically enriched for terms such as spermatid development and flagellar assembly.Furthermore,we showed that SPEF2 interacts with radial spoke head component 9(RSPH9)and IFT20 in vitro,which are well-studied components of radial spokes or intra-flagellar transport and are essential for flagellar assembly.These results provide a rich resource for further investigation into the molecular mechanism underlying the role that SPEF2 plays in sperm tail development and could provide a theoretical basis for gene therapy in SPEF2 mutant patients in the future.
基金The work was supported by Graduate Independent Innovation Project Fund of Central South University(2020zzts229).
文摘Sperm morphology was once believed as one of the most predictive indicators of pregnancy outcome in assisted reproductive technology(ART).However,the impact of teratozoospermia on in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI)outcomes and its offspring remains inconclusive.In order to evaluate the influence of teratozoospermia on pregnancy outcome and newborn status after IVF and ICSI,a retrospective study was conducted.This was a matched case-control study that included 2202 IVF cycles and 2574 ICSI cycles and was conducted at the Reproductive and Genetic Hospital of CITIC-Xiangya in Changsha,China,from June 2013 to June 2018.Patients were divided into two groups based on sperm morphology:teratozoospermia and normal sperm group.The pregnancy outcome and newborn outcome were analyzed.The results indicated that couples with teratozoospermia had a significantly lower optimal embryo rate compared to those with normal sperm morphology in IVF(P=0.007),while there were no statistically significant differences between the two groups in terms of the fertilization rate,cleavage rate,implantation rate,and pregnancy rate(all P>0.05).Additionally,teratozoospermia was associated with lower infant birth weight in multiple births after IVF.With regard to ICSI,there was no significant difference in both pregnancy outcome and newborn outcome between the teratozoospermia and normal groups(both P>0.05).Furthermore,no increase in the risk of birth defects occurred in the teratozoospermia group after IVF/ICSI.Consequently,we believe that teratozoospermia has limited predictive value for pregnancy outcomes in IVF/ICSI,and has little impact on the resulting offspring if multiple pregnancy is avoided.
基金This study was supported by grants from the National Key Research&Developmental Program of China(2018YFC1004901)National Natural Science Foundation of China(81771645 and 81971447)+2 种基金Hunan Provincial Natural Science Foundation of China(2019JJ51006)the Key Grant of Prevention and Treatment of Birth Defect from Hunan Province(2019SK1012)the Scientific Research Foundation of Reproductive and Genetic Hospital of CITIC-Xiangya grants(YNXM-201912).
文摘Diminished ovarian reserve(DOR)is a disorder of ovarian function in which the ovary loses its normal reproductive potential,including decreasing oocyte quantity and quality.The disorder is associated with infertility in women of reproductive age with regular menses and poor response to controlled ovarian hyperstimulation(Broekmans et al.,2009).
基金the National Natural Science Foundation for Young Scholars of China,No.82201771National Natural Science Foundation of China,No.32270912+2 种基金Natural Science Foundation of Changsha,No.kq2202491Research Grant of CITIC-Xiangya,No.YNXM202109 and No.YNXM202115Hunan Provincial Grant for Innovative Province Construction,No.2019SK4012。
文摘BACKGROUND Spermatogonial stem cells(SSCs)are the origin of male spermatogenesis,which can reconstruct germ cell lineage in mice.However,the application of SSCs for male fertility restoration is hindered due to the unclear mechanisms of proliferation and self-renewal in humans.AIM To investigate the role and mechanism of SPOC domain-containing protein 1(SPOCD1)in human SSC proliferation.METHODS We analyzed publicly available human testis single-cell RNA sequencing(RNAseq)data and found that SPOCD1 is predominantly expressed in SSCs in the early developmental stages.Small interfering RNA was applied to suppress SPOCD1 expression to detect the impacts of SPOCD1 inhibition on SSC proliferation and apoptosis.Subsequently,we explored the target genes of SPOCD1 using RNA-seq and confirmed their role by restoring the expression of the target genes.In addition,we examined SPOCD1 expression in some non-obstructive azoospermia(NOA)patients to explore the correlation between SPOCD1 and NOA.RESULTS The uniform manifold approximation and projection clustering and pseudotime analysis showed that SPOCD1 was highly expressed in the early stages of SSC,and immunohistological results showed that SPOCD1 was mainly localized in glial cell line-derived neurotrophic factor family receptor alpha-1 positive SSCs.SPOCD1 knockdown significantly inhibited cell proliferation and promoted apoptosis.RNA-seq results showed that SPOCD1 knockdown significantly downregulated genes such as adenylate kinase 4(AK4).Overexpression of AK4 in SPOCD1 knockdown cells partially reversed the phenotypic changes,indicating that AK4 is a functional target gene of SPOCD1.In addition,we found a significant downregulation of SPOCD1 expression in some NOA patients,suggesting that the downregulation of SPOCD1 may be relevant for NOA.CONCLUSION Our study broadens the understanding of human SSC fate determination and may offer new theories on the etiology of male infertility.
基金support from the National Key Research and Developmental Program of China (No.2018YFC1003603)the National Natural Science Foundation of China (No.81971445).
文摘Congenital bilateral absence of the vas deferens(CBAVD)is observed in 1%–2%of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator(CFTR)mutations.CFTR is one of the most well-known genes related to male fertility.The frequency of CFTR mutations or impaired CFTR expression is increased in men with nonobstructive azoospermia(NOA).CFTR mutations are highly polymorphic and have established ethnic specificity.Compared with F508Del in Caucasians,the p.G970D mutation is reported to be the most frequent CFTR mutation in Chinese patients with cystic fibrosis.However,whether p.G970D participates in male infertility remains unknown.Herein,a loss-of-function CFTR p.G970D missense mutation was identified in a patient with CBAVD and NOA.Subsequent retrospective analysis of 122 Chinese patients with CBAVD showed that the mutation is a common pathogenic mutation(4.1%,5/122),excluding polymorphic sites.Furthermore,we generated model cell lines derived from mouse testes harboring the homozygous Cftr p.G965D mutation equivalent to the CFTR variant in patients.The Cftr p.G965D mutation may be lethal in spermatogonial stem cells and spermatogonia and affect the proliferation of spermatocytes and Sertoli cells.In spermatocyte GC-2(spd)ts(GC2)Cftr p.G965D cells,RNA splicing variants were detected and CFTR expression decreased,which may contribute to the phenotypes associated with impaired spermatogenesis.Thus,this study indicated that the CFTR p.G970D missense mutation might be a pathogenic mutation for CBAVD in Chinese males and associated with impaired spermatogenesis by affecting the proliferation of germ cells.
文摘Dear Editor, There are two main causes of severe asthenozoospermia： ultrastructural defects （genetically inherited and congenital defects） of the sperm flagellum and necrozoospermia or sperm degeneration secondary to other pathological changes （see review by Ortega et al2）. Genetic-related absolute asthenozoospermia includes primary ciliary dyskinesia and dysplasia of the fibrous sheath （DFS）, which result in 100% （or nearly） immotile spermatozoa.
基金supported by grants from the National Key Research and Development Program of China(2022YFC2702604)the National Natural Science Foundation of China(82171608,82201773,and 81971447)+1 种基金the China Postdoctoral Science Foundation(2022M711119)the Scientific Research Foundation of the Health Committee of Hunan Province(B202301039323 and B202301039518).
文摘Male infertility is a major reproductive disorder,which is clinically characterized by highly heterogeneous phenotypes of abnormal sperm count or quality.To date,five male patients with biallelic loss-of-function(LOF)variants of PARN-like ribonuclease domain-containing exonuclease 1(PNLDC1)have been reported to experience infertility with nonobstructive azoospermia.The aim of this study was to identify the genetic cause of male infertility with oligo-astheno-teratozoospermia(OAT)in a patient from a Chinese Han family.Whole-exome and Sanger sequencing analyses identified a homozygous LOF variant(NM_173516.2,c.l42C>T,p.Gln48Ter)in PNLDC1.Hematoxylin and eosin staining revealed that the spermatozoa of the patient with OAT had an irregular head phenotype,including microcephaly,head tapering,and globozoospermia.Consistently,peanut agglutinin staining of the spermatozoa revealed a complete or partial loss of the acrosome.Furthermore,the disomy rate of chromosomes in the patient’s spermatozoa was significantly increased compared with that of a fertile control sample.We reported an LOF variant of the PNLDC1 gene responsible for OAT.
基金financial supported by National Natural Science Foundationof China(No.82001634)the China Postdoctoral Science Foundation(No.2019M661521).
文摘Pericentric inversion of chromosome 9(inv[9])is a common chromosomal structural variant,but its impact on clinical outcomesremains debated.The screening criteria of sperm banks are rarely mentioned to individuals with inv(9).In this study,we evaluatedthe fertility of sperm donors with inv(9)who met eligibility criteria for sperm banks(inv[9]-eligible donors).From March 2004 toMay 2022,chromosomal analysis of 16124 sperm donors at CITIC-Xiangya Human Sperm Bank in Hunan Province(Changsha,China)found that 251(1.6%)had chromosome variations,with inv(9)being the most prevalent at 1.1%.All 169 inv(9)-eligibledonors were contacted to collect fertility outcome data,along with 206 eligible donors without inv(9)as controls.In addition,semen samples from inv(9)-eligible donors and eligible donors underwent assessments of sperm fluorescence in situ hybridization(FISH),mitochondrial membrane potential,DNA fragmentation index,acrosome integrity,reactive oxygen species(ROS),andsperm morphology.Results showed that inv(9)did not significantly increase reproductive risks overall.Despite detecting ROSlevel differences,the clinical impact may be insignificant.This study provides new data on the inv(9)population that can serveas a valuable reference for decision-making by sperm banks as well as for genetic counseling and clinical guidance for individualscarrying inv(9)variant.
