Homozygous CCDC146 mutation causes oligoasthenoteratozoospermia in humans and mice

Infertility represents a significant health concern,with sperm quantity and quality being crucial determinants of male fertility.Oligoasthenoteratozoospermia(OAT)is characterized by reduced sperm motility,lower sperm concentration,and morphological abnormalities in sperm heads and flagella.Although variants in several genes have been implicated in OAT,its genetic etiologies and pathogenetic mechanisms remain inadequately understood.In this study,we identified a homozygous nonsense mutation(c.916C>T,p.Arg306*)in the coiled-coil domain containing 146(CCDC146)gene in an infertile male patient with OAT.This mutation resulted in the production of a truncated CCDC146 protein(amino acids 1-305),retaining only two out of five coiled-coil domains.To validate the pathogenicity of the CCDC146 mutation,we generated a mouse model(Ccdc146^(mut/mut))with a similar mutation to that of the patient.Consistently,the Ccdc146mut/mut mice exhibited infertility,characterized by significantly reduced sperm counts,diminished motility,and multiple defects in sperm heads and flagella.Furthermore,the levels of axonemal proteins,including DNAH17,DNAH1,and SPAG6,were significantly reduced in the sperm of Ccdc146^(mut/mut) mice.Additionally,both human and mouse CCDC146 interacted with intraflagellar transport protein 20(IFT20),but this interaction was lost in the mutated versions,leading to the degradation of IFT20.This study identified a novel deleterious homozygous nonsense mutation in CCDC146 that causes male infertility,potentially by disrupting axonemal protein transportation.These findings offer valuable insights for genetic counseling and understanding the mechanisms underlying CCDC146 mutant-associated infertility in human males.

Male idiopathic oligoasthenoteratozoospermia

Idiopathic oligoasthenoteratozoospermia （iOAT） affects approximately 30% of all infertile men. This mini-review discussed recent data in this field. Age, non-inflammatory functional alterations in post-testicular organs, infective agents （Chlamydia trachomatis, herpes virus and adeno-associated viruses）, alterations in gamete genome, mitochondrial alterations, environmental pollutants and ＂subde＂ hormonal alterations are all considered possible causes of iOAT. Increase of reactive oxygen species in tubules and in seminal plasma and of apoptosis are reputed to affect sperm concentration, motility and morphology, iOAT is commonly diagnosed by exclusion, nevertheless spectral traces of the main testicular artery may be used as a diagnostic tool for iOAT. The following can be considered therapies for iOAT： 1） tamoxifen citrate （20 mg/d） ＋ testosterone undecanoate （120 mg/d） （pregnancy rate per couple/month [prcm]： 3.8%）; 2） folic acid （66 mg/d） ＋ zinc sulfate （5 mg/d）; 3） L-carnitine （2 g/d） alone or in combination with acetyl-L- canitine （1 g/d） （prcm： 2.3%）; and 4） both camitines ＋ one 30 mg cinnoxicam suppository every 4 days （prcm： 8.5%）. Alpha-blocking drugs improved sperm concentration but not morphology, motility or pregnancy rate. Tranilast （300 mg/d） increased sperm parameters and pregnancy rates in an initial uncontrolled study. Its efficacy on sperm concentration （but not on sperm motility, morphology or prcm） was confirmed in subsequent published reports. The efficacy of tamoxifen ＋ testosterone undecanoate, tamoxifen alone, and recombinant follicle-stimulating hormone is still a matter for discussion. （Asian J Andro12006 Mar; 8： 143-157）

Antioxidant treatment for oligoasthenoteratozoospermia and varicocele:a DBPC trial to evaluate the impact of age and body mass index

Oxidative stress is one of the main mechanisms responsible for male infertility.Various conditions such as varicocele,obesity,advanced age,and lifestyle can lead to an increase in reactive oxygen species,causing an oxidative imbalance in the reproductiveenvironment.Spermatozoa are sensitive to reactive oxygen species and require energy to carry out their main function of fertilizingthe egg.Excessive reactive oxygen species can affect sperm metabolism,leading to immobility,impaired acrosome reaction,and celldeath,thereby impairing reproductive success.This double-blind randomized study evaluated the effect of supplementation withL-carnitine,acetyl-L-carnitine,vitamins,and other nutrients on semen quality in 104 infertile patients with or without varicocele,while also investigating the impact of factors such as obesity and advanced age on treatment.Sperm concentration significantlyincreased in the supplemented group compared to the placebo group(P=0.0186).Total sperm count also significantly increasedin the supplemented group(P=0.0117),as did sperm motility(P=0.0120).The treatment had a positive effect on patients upto 35 years of age in terms of sperm concentration(P=0.0352),while a body mass index(BMI)above 25 kg m^(−2) had a negativeeffect on sperm concentration(P=0.0110).Results were not showing a net benefit in stratifying patients in accordance withtheir BMI since sperm quality increase was not affected by this parameter.In conclusion,antioxidant supplementation may bebeneficial for infertile patients and has a more positive effect on younger patients with a normal weight.

Deficiency of MFSD6L, an acrosome membrane protein, causes oligoasthenoteratozoospermia in humans and mice

Oligoasthenoteratozoospermia is an important factor affecting male fertility and has been found to be associated with genetic factors.However,there are stll a proportion of oligoasthenoteratozoospermia cases that cannot be explained by known pathogenic genetic variants.Here,we perform genetic analyses and identify bi-allelic loss-of-function variants of MFSD6L from an oligoasthenoteratozoospermia-affected family.Mfsd6l knock-out male mice also present male subfertility with reduced sperm concentration,motility,and deformed acrosomes.Further mechanistic analyses reveal that MFsD6L,as an acrosome membrane protein,plays an important role in the formation of acrosome by interacting with the inner acrosomal membrane protein SPACA1.Moreover,poor embryonic development is consistently observed after intracytoplasmic sperm injection treatment using spermatozoa from the MFSD6L-deficient man and male mice.Collectively,our findings reveal that MFSD6L is required for the anchoring of sperm acrosome and head shaping.The deficiency of MFsD6L affects male fertility and causes oligoasthenoter-atozoospermia in humans and mice.

关于少弱畸形精子症蛋白组学的研究进展

少弱畸形精子症(OAT)是导致男性不育的常见疾病,特征为精子数量减少及活力降低。该疾病的发病机制复杂,涉及生活方式、遗传异常、环境因素等。近年来,精子蛋白组学的研究取得显著进展,尤其是在OAT患者中的应用。本文综述了OAT患者精子及精浆蛋白组的研究进展,描述了与精子活力和数量相关的潜在差异蛋白,旨在深入理解OAT的病理机制,提高诊断准确性,并探索新的生物标记物。

A novel homozygous frameshift mutation in MNS1 associated with severe oligoasthenoteratozoospermia in humans

Oligoasthenoteratozoospermia(OAT)refers to the combination of various sperm abnormalities,including a decreased sperm count,reduced motility,and abnormal sperm morphology.Only a few genetic causes have been shown to be associated with OAT.Herein,we identified a novel homozygous frameshift mutation in meiosis-specific nuclear structural 1(MNS1;NM_018365:c.603_604insG:p.Lys202Glufs*6)by whole-exome sequencing in an OAT proband from a consanguineous Chinese family.Subsequent variant screening identified four additional heterozygous MNS1 variants in 6/219 infertile individuals with oligoasthenospermia,but no MNS1 variants were observed among 223 fertile controls.Immunostaining analysis showed MNS1 to be normally located in the whole-sperm flagella,but was absent in the proband's sperm.Expression analysis by Western blot also confirmed that MNS1 was absent in the proband's sperm.Abnormal flagellum morphology and ultrastructural disturbances in outer doublet microtubules were observed in the proband's sperm.A total of three intracytoplasmic sperm injection cycles were carried out for the proband's wife,but they all failed to lead to a successful pregnancy.Overall,this is the first study to report a loss-of-function mutation in MNS1 causing OAT in a Han Chinese patient.

Evaluation on Antisperm Antibody in Infertile Men with Oligoasthenoteratozoospermia

To evaluate the level of antisperm antibody （ASA） in infertile men with oligoasthenoteratozoospermia （OAT）. Methods Forty-six infertile men with OAT were enrolled into this study. Sperm samples were screened by the direct immunobead test for ASA type IgG and lgA according to the WHO laboratory manual. Results Of the 46 patients with OAT assessed for ASA-IgG, 2 had immunobead binding which were 22% and 27%, respectively, and the sub-positive rate was 4.3%. No case had the clinical positive level according to the WHO criteria （ ≥ 50% of the motile sperm with immunobead binding）. ASA-IgA was not detected in all cases. Conclusion A significant incidence or high level of ASA could not be found in infertile men with OAT, which suggests that ASA is not associated with the pathogenesis of infertile men with OAT.

Evaluation of Glutathione Peroxidase Enzymatic Activity in Seminal Plasma of Patients Treated at the Institute Pasteur in Cote d’Ivoire

Glutathione peroxidase (GPx) is an antioxidant that plays an important role in the maintenance of male fertility. The aim of this study was to compare the profile of enzymatic activity of glutathione peroxidase in the seminal plasma of normozoosperm and those of pathological sperm. Thus, the activity of glutathione peroxidase was determined in the seminal plasma of 20 normozoosperms, 9 azoosperms and 31 oligoasthenoteratozoosperms. It was 37.58 ± 3.14 U/L in normozoosperms, 39.39 ± 2.27 U/L in oligoasthenoteratozoosperms, and 29.77 ± 2.62 U/L in azoosperms. The mean GPx enzyme activity of normozoosperms did not differ significantly from that of oligoasthenoteratozoosperms and azoosperms. In contrast, comparison of enzyme activity between abnormal sperms gave a significant difference. This study showed that glutathione peroxidase enzymatic activity is not related to sperm quality.

Varicocele and Spermogram Abnormality: About 264 Cases at the Fertilia Medical Clinic in Bamako

Aims: To determine the impact of varicocele on the result of the spermogram in a group of hypo-fertile or infertile men. Patients and Method: This was a cross-sectional, descriptive study concerning 264 men collected between February 2020 and March 2022 at the Fertilia medical clinic in Bamako. The study population consisted of consenting male subjects presenting for infertility. All of our patients had already done a spermogram, they then benefited from a scrotal Doppler ultrasound. The scrotal echo-doppler was completed by an abdominal and pelvic echography in search of a possible etiology, in particular a renal cancer, a thrombosis of the renal or spermatic vein, a retroperitoneal mass or a nutcraker syndrome. Testicular volume was measured on ultrasound according to Lambert’s formula (length × width × height × 0.71). A volume = 16 ml was considered normal. Results: 264 patients were recruited in ours. The average age was 33 years with extremes ranging from 25 to 65 years. 149 patients or 56.44% were between 30 and 45 years old. 213 patients or 80.68% had primary infertility. All the patients, i.e. 100%, had a sperm anomaly and dilation of the pampiniform plexus veins. The most common sperm abnormalities were oligoasthenoteratozoospermia, which concerned 214 patients, or 81%, followed by azoospermia (40 patients, or 15%) and oligospermia (10 patients, or 4%). 213 patients or 80.68% had a Hirshen grade II varicocele, 45 patients or 17% had grade I and 06 patients or 2.27% had grade III. Among the 6 patients, 04 had left testicular atrophy and 2 bilateral atrophy. The venous reflux was more accentuated on the left than on the right. Conclusion: Varicocele is significantly found in men with infertility. Sperm alterations concern both the concentration of spermatozoa, their mobility and their vitality. Even if the mechanism of alteration of the sperm parameters is not well elucidated, our study made it possible to make the link between the dilation of the veins of the pampiniform plexus and the sperm abnormalities mentioned above. It should be recognized that varicocelectomy has allowed a significant improvement in the quality of sperm with the obtaining of a certain number of natural pregnancies and those resulting from medically assisted procreation.

贺菊乔教授辨治特发性少弱畸形精子症经验

介绍了贺菊乔教授以"肾-天癸-肾子轴"论治特发性少弱畸形精子症的临床经验,提出"肾-天癸-肾子轴"为肾主生殖功能的直接体现,指明其与脏腑调控的相关性,揭示了其与"下丘脑-腺垂体-睾丸轴"的相通之处,并介绍了贺菊乔教授治疗特发性少弱畸形精子症的临证思路和效验方药。

13号染色体重排所致严重少弱畸形精子症患者生精细胞减数分裂分析

目的:探讨13号染色体长臂相互缺失所致严重少弱畸形精子症患者生精阻滞的可能机制。方法:对1例13号染色体异常的严重少弱畸形精子症患者血液样本进行全基因组的寡核苷酸微阵列比较基因组杂交分析,以确定受累染色体的断裂点或基因缺失。对患者生殖细胞进行多色荧光原位杂交分析,以观察初级精母细胞13号染色体配对的情况。结果:寡核苷酸微阵列比较基因组杂交技术显示在13q12.3上有连续4个探针的缺失(A_16_P19757882,A_16_P02744617,A_14_P108858和A_16_P02744687),覆盖59.93 kb,位于基因FLT1和POMP之间,没有注释基因存在。初级精母细胞的减数分裂分析显示同源13号染色体配对错误,13q14和13qter信号彼此分离。结论:染色体重排导致的精子发生阻滞可能是由于生殖细胞第一次减数分裂同源染色体配对错误导致的。

芳香化酶抑制剂在男性不育治疗中的应用

男性不育的病因复杂多样,且多被归为特发性因素。许多特发性男性不育症患者均表现为体内睾酮(T)水平下降、雌二醇(E2)水平上升、T/E2失调和生精功能障碍等。芳香化酶在男性体内雄激素转化为雌激素的过程中发挥关键作用。以来曲唑为代表的芳香化酶抑制剂可有效抑制芳香化酶活性,从而降低E2水平、提升T与促性腺激素水平,提高T/E2,改善生精功能,对提高不育症自然妊娠率与人工辅助生殖的成功率具有重要意义。大样本的随机对照临床试验将进一步明确其在男性不育症治疗中的地位,并有望成为男性不育症的常规治疗药物。

少弱畸形精子优选后体外生存时间与宫腔内人工授精妊娠率的对比研究

为比较研究少弱畸形精子优选后体外生存时间与宫腔内人工授精(IUI)妊娠率的关系。选择因男方患少弱畸形精子症致不孕者。以改良F10培养液为精子培养液,采用上泳分离法体外处理精子后,置36.5℃二氧化碳培养箱中,分别于0,9,12,24,36,48h观察其精子活率和活力,活率≥50％者为阳性,活率<50％者为阴性。结果以生存时间(SST)表示。宫腔内人工授精按常规操作进行。共治疗60例,137个周期,妊娠19例,总妊娠率为31.7％(19/60),周期妊娠率为13.8％(19/137)。其中 SST 9h阳性5例,无一例妊娠,12h阳性10例,2例妊娠(20％),24h阳性16例,4例妊娠(25％),36h阳性29例,13例妊娠(44.8％),48h精子生存试验结果显示阴性,优选后精子体外生存时间对预测IUI妊娠率有重要临床价值,即SST为36h,人工授精妊娠率明显增高。

卵胞浆内单精子注射治疗严重男性因素不育症428个周期的临床分析

目的分析卵胞浆内单精子显微注射技术(intracytoplasmic sperm injection,ICSI)在治疗严重男性因素不育症时的有效性和安全性。方法回顾分析2009年1月至2010年12月间在本院因严重男性不育症行ICSI助孕的428例患者的数据,其中245例梗阻性无精子症组采用经皮附睾穿刺抽吸术取得的附睾精子;183例严重少弱畸精子症组采用的是射出精液中的精子。并且收集患者的年龄、体重指数、基础卵泡刺激素(FSH)、移植胚胎数目、受精率、优胚率、取消率、妊娠率、着床率、流产率、宫外孕发生率、妊娠出生率、移植出生率、出生异常率等资料。结果梗阻性无精子症组受精率(84.1±17.1)%,着床率45.2%,移植出生率5 9.1%,流产率10.3%,出生异常率3.5%。严重少弱畸精子症组受精率(79.9±20.9)%,着床率3 7.8%,移植出生率48.2%,流产率15.2%,出生异常率3.1%。两组均获得了满意的临床结局,但两组的受精率、着床率、移植出生率及流产率均存在统计学差异。结论本研究结果提示梗阻性无精子组的受精率,着床率及移植出生率较严重少弱畸精子症组均明显升高,且术后流产率较低,提示梗阻性无精子症患者经附睾穿刺行ICSI较严重少弱畸精子症患者可以获得更好的临床结局。但此技术的安全性,依然需要更多的研究。

特发性无精子症及少弱精子症患者Y染色体AZF微缺失筛查分析

目的 探讨特发性无精子症及少弱精子症不育男性与Y染色体AZF微缺失的关系。方法 用双重PCR技术对 6 3例患者 (无精子症 4 1例 ,少弱精子症 14例 ,严重少精子症 8例 )进行Y染色体AZFa、AZFb、AZFc、SRY的微缺失筛查。同时对 2 6例无精子症患者行睾丸活检、组织学评估。结果 6 3例中AZF微缺失 7例 ,缺失率为 11 1%。其中无精子症 5例 ,严重少精子症 2例。AZFc缺失 4例 ,AZFb缺失 2例 ,AZFb +AZFc缺失 1例 ,未发现AZFa区缺失。 6 3例及对照组 30例SRY基因扩增均阳性。 2 6例无精子症患者行睾丸活检、组织学检查 ,无 1例精子发生正常。结论 Y染色体微缺失 ,特别是AZFc区DAZ基因的微缺失 ,是引起无精子和严重少弱精子等生精障碍而致男性不育较为重要的遗传学因素。

来曲唑联合维生素E对特发性少弱畸精子症患者精液质量、血清睾酮和雌二醇水平及氧化应激损伤的影响

目的采用来曲唑联合维生素E治疗特发性少弱畸精子症(iOAT)患者,观察其对患者精液质量、精子DNA碎片指数(DFI)、血清睾酮(T)和雌二醇(E_(2))水平及氧化应激水平的影响。方法选取2019年1月至2021年1月联勤保障部队解放军第960医院生殖医学科收治的196例iOAT患者作为研究对象。随机分为试验组(n=100)和对照组(n=96)。试验组采用维生素E软胶囊联合来曲唑治疗,对照组采用单纯维生素E软胶囊治疗。对两组精液常规参数、精子DFI、血清T和E_(2)水平、精液氧化应激水平进行比较。结果治疗后,试验组精子浓度、前向运动精子百分率、正常形态精子百分率显著高于对照组,DFI显著低于对照组(P<0.05);试验组T、T/E_(2)水平显著高于对照组,E_(2)水平显著低于对照组(P<0.05);试验组活性氧(ROS)和丙二醛(MDA)水平显著低于对照组,超氧化物歧化酶(SOD)水平显著高于对照组(P<0.05)。结论对iOAT患者采用来曲唑联合维生素E软胶囊治疗可显著改善患者精液质量,调节血清中激素水平,降低氧化应激水平。

尿素加盐酸胍裂解精子效果分析

目的探讨尿素加盐酸胍裂解精子的有效性。方法将80例少弱畸精子症患者按随机数字表法分成两组,每组40例,采集精液标本提取精子,分别应用尿素加盐酸胍裂解法或尿素裂解法对精子进行裂解。结果尿素加盐酸胍组得到蛋白质浓度、蛋白质点分别为(1.25±0.13)μg/μl、(864±52)个,均明显高于尿素组的(0.74±0.15)μg/μl、(718±63)个(P<0.05)。结论尿素加盐酸胍裂解法能得到更丰富、更全面的精子蛋白质种类,并提高蛋白质含量,使双向电泳图谱更加完善,更有利于寻找差异蛋白质。

精子变形相关基因在少弱畸形精子症发生中的作用

不孕不育是一种常见的疾病,影响大约10%~15%的育龄夫妇,其中男性因素占一半.通过精子发生产生形态与功能正常的精子是人类繁衍的基础.精子发生包括有丝分裂、减数分裂和精子变形三个阶段.其中,精子变形是雄性生殖细胞发育所特有的阶段,其相关基因的异常与少弱畸形精子症(oligoasthenoteratozoospermia, OAT)这一男性不育的常见类型密切相关.近年常有精子变形相关基因异常导致人或小鼠出现OAT表型的报道.但目前对于精子变形相关基因在OAT发生中的作用尚未有系统性的归纳.本文围绕精子变形中的关键事件及部分代表性的相关基因在OAT发生中的作用进行综述,希望对OAT患者的生育咨询、治疗和助孕提供更好的帮助.

辅酶Q10治疗特发性少弱畸形精子症有效性与安全性的Meta分析

目的系统评价抗氧化剂辅酶Q10(coenzyme Q10,Co Q10)治疗特发性少弱畸形精子症(idiopathic oligoasthenoteratozoospermia,i OAT)的有效性及安全性。方法计算机检索Pub Med、MEDLINE、EMbase、The Cochrane Library、CBM、CNKI、VIP和Wan Fang Data数据库,搜集Co Q10治疗i OAT的相关随机对照试验(RCT),检索时限均为从建库至2016年5月31日。由2位研究者独立进行文献筛选、资料提取,并评价纳入研究的偏倚风险后,采用Rev Man 5.3软件进行Meta分析。结果最终纳入7个RCT,共803例患者。Meta分析结果显示:与对照组相比,Co Q10组可明显提高i OAT患者的精子浓度[MD=3.37,95%CI(0.68,6.05),P=0.01]、a级精子百分率[MD=5.06,95%CI(3.84,6.28),P<0.000 01]、(a+b)级精子百分率[MD=7.72,95%CI(4.19,11.26),P<0.000 1]、正常形态精子百分率[MD=1.89,95%CI(0.63,3.16),P=0.003]和精液中Co Q10的水平[MD=40.02,95%CI(24.73,55.31),P<0.000 01];但不能明显改善血浆性激素水平[FSH:MD=–3.48,95%CI(–5.17,–1.79),P<0.000 1;LH:MD=–3.23,95%CI(–7.55,1.08),P=0.14;T:MD=0.45,95%CI(–3.31,4.20),P=0.82];此外,两组均未出现与药物相关的明显不良反应。结论抗氧化剂Co Q10可明显提高特发性男性不育症患者的精子浓度和精子活力,未见明显不良反应,但由于纳入研究存在较大的偏倚风险,证据强度较低,本研究结论尚需进一步开展设计严格的临床对照试验进行验证。

严重少精子症合并罕见染色体重排——1个超数新着丝粒环状染色体和1个平衡缺失染色体

目的:报道1例少弱畸精子症不育男性合并罕见的染色体重排。方法:进行GTG-显带,CBG-显带和荧光原位杂交分析染色体核型。用微阵列比较基因组杂交和AZF微缺失检测核型拷贝数变化。结果:分裂相中存在1条线性标记染色体和1个环状标记染色体,它们都源于13号染色体。且环形染色体不带有α-卫星DNA;Y染色体AZF微缺失检测结果正常。微阵列比较基因组杂交未观察到13号染色体拷贝数缺失。结论:本研究首次报告了1例染色体重排导致的平衡核型合并严重少弱畸精子症。其可能原因是染色体重排干扰了减数分裂I期染色体联会重组过程,导致减数分裂阻滞在精母细胞阶段。但这还有待于进一步的实验证实。