BACKGROUND & AIMS: :Diabetes mellitus in human and animal models has been correlated with low sperm count, testicular abnormalities, high levels of germ cell death, and oxidative stress. In this study, we focus on three questions: (1) Is germ cell apoptosis stage-specific in diabetic male rats? (2) Could ascorbic acid (AA) reverse oxidative and histological damage and restore testicular dysfunction? (3) Could AA treatment restore fertility parameters in diabetic rats? Adult Sprague-Dawley rats were divided into four groups: control, diabetic, control plus AA, and diabetic plus AA. Seminiferous tubules underwent severe histological damage, together with a change in frequency of some stages of the seminiferous cycle, and germ cell apoptosis was increased in a stage-dependent manner in diabetic rats. We found a significant decrease in testosterone and higher levels of lipid peroxidation in diabetic rats when compared with controls. A major finding was that AA reversed the histological damage and peroxidation levels to control levels in diabetic rats, but testosterone levels remained unchanged. The pregnancy rate was decreased in females that mated with diabetic rats and those treated with AA, but the litter size was only reduced in the second case. Interestingly, spermatozoa from diabetic and AA-treated rats showed reduced motility and hyperactivation, but only diabetic rats had higher levels of apoptosis when compared with controls. These results suggest that treatment with AA reverses testicular damage in diabetic rats but is insufficient to restore testosterone levels, sperm motility, and fertility in a rat model.

背景与目标： 人和动物模型中的糖尿病与精子数量低，睾丸异常，生殖细胞死亡水平高和氧化应激有关。在这项研究中，我们关注三个问题：（1）糖尿病雄性大鼠生殖细胞凋亡是否具有阶段特异性？ （2）抗坏血酸（AA）能逆转氧化和组织损伤并恢复睾丸功能障碍吗？ （3）AA治疗能恢复糖尿病大鼠的生育能力吗？将成年的Sprague-Dawley大鼠分为四组：对照组，糖尿病，对照组加AA和糖尿病加AA。曲细精管经历了严重的组织学损伤，并伴有曲细精循环某些阶段的频率变化，并且在糖尿病大鼠中生殖细胞凋亡以阶段依赖性的方式增加。我们发现与对照组相比，糖尿病大鼠睾丸激素显着降低，脂质过氧化水平更高。一个主要发现是，AA使糖尿病大鼠的组织学损伤和过氧化水平逆转至对照水平，但睾丸激素水平保持不变。与糖尿病大鼠交配的雌性和接受AA治疗的雌性，其妊娠率降低，但仅在第二种情况下，其产仔数减少。有趣的是，来自糖尿病和AA治疗的大鼠的精子显示出降低的运动性和过度激活，但是与对照组相比，只有糖尿病大鼠具有更高的细胞凋亡水平。这些结果表明，AA治疗可以逆转糖尿病大鼠的睾丸损伤，但不足以恢复大鼠模型中的睾丸激素水平，精子活动性和受精能力。

BACKGROUND & AIMS: :Maintaining the integrity of spermatogenic stem cells is essential to transfer genetic information to a descendant. However, knowledge of maintenance of genetic stability in stem cells is still limited. RAD18 is critical for postreplication repair through mono- and multi-ubiquitination of proliferating cell nuclear antigen (PCNA) to maintain genomic stability. Mammalian RAD18 is highly expressed in the spermatocytes and the nuclei of a few spermatogonia in adult mice. To elucidate the physiological function of RAD18, we analyzed a phenotype of Rad18-/- mice. The mice were born and appeared to grow normally. Although the mice were fertile, fertility and testis weight decreased with age. Histological examination revealed normal spermatogenesis in almost all seminiferous tubules in Rad18-/- testes at 2 months old, and abnormal sperm could not be detected in the epididymis. However, 25% of the tubules lost almost all germ cells at 12 months. The seminiferous tubules frequently retained only late differentiated phase germ cells, suggesting that the exhaustion of spermatogonial stem cells leads to the loss of all germ cells in the seminiferous tubules. Wild-type germ cells were successfully transplanted into and colonized in the seminiferous tubules of aged Rad18-/- mice, indicating that Sertoli cells have a normal supportive function even in aged testes. We conclude that RAD18 is intrinsically required for the long-term maintenance of spermatogenesis.

背景与目标： ：维持生精干细胞的完整性对于将遗传信息传递给后代至关重要。但是，维持干细胞遗传稳定性的知识仍然有限。 RAD18通过增殖细胞核抗原（PCNA）的单泛素化和多泛素化来维持基因组稳定性，对于复制后修复至关重要。哺乳动物RAD18在成年小鼠的精细胞和少数精原细胞的核中高度表达。为了阐明RAD18的生理功能，我们分析了Rad18-/-小鼠的表型。小鼠出生并且看上去正常生长。尽管小鼠具有生育能力，但其生育力和睾丸重量会随着年龄的增长而下降。组织学检查显示，在两个月大的Rad18-/-睾丸中，几乎所有曲细精管中的精子发生均正常，并且在附睾中未检测到异常的精子。但是，在12个月时，有25％的肾小管几乎失去了所有生殖细胞。曲细精管经常仅保留晚期分化期的生殖细胞，这表明精原干细胞的耗尽导致了曲细精管中所有生殖细胞的丢失。野生型生殖细胞已成功移植到老年Rad18-/-小鼠的生精小管中并在其中繁殖，这表明Sertoli细胞即使在老年睾丸中也具有正常的支持功能。我们得出结论，RAD18是精子发生长期维持的内在要求。

BACKGROUND & AIMS: BACKGROUND:Histone to protamine exchange, in man, is only 80% complete and spermatozoal histones are highly acetylated suggesting that their associated genes may be involved in early gene expression in the embryo. METHODS:Using in situ hybridization and immunocytochemistry, we analyzed expression of protamine-1 and protamine-2, and histone H4 specifically acetylated at lysine 5 (H4K5ac), 8 (H4K8ac), 12 (H4K12ac) and 16 (H4K16ac) in human (n = 22) and marmoset (n = 6) testes and ejaculates. RESULTS:Protamine-1 and protamine-2 mRNA was present in round and elongating spermatids. All antibodies against acetylated histones revealed positive signals in these cells. Human spermatogonia showed positive signals for H4K8ac and H4K16ac, whereas marmoset spermatogonia were positive for H4K8ac and H4K12ac. In man, H4K16ac already displayed a positive immunoreaction with pachytene spermatocytes, starting at stage III of the seminiferous epithelial cycle. All antibodies showed positive immunostaining in ejaculated spermatozoa of both species. CONCLUSIONS:The common marmoset represents a suitable animal model for studies on nuclear protein expression during human spermatogenesis. The two species exhibit a similar organization of seminiferous epithelium and an identical expression pattern of protamine-1 and protamine-2 mRNA in round and elongating spermatids. The presence of specifically acetylated histones H4 in testicular spermatids and ejaculated spermatozoa demonstrates an incomplete histone to protamine exchange in both species. As acetylated histones are known to be associated with genes involved in gene expression, the common marmoset may, in future, be used as a model for investigations on early embryo development which, in man, are not possible for ethical reasons.

背景与目标： 背景：在人类中，组蛋白与鱼精蛋白的交换仅完成了80％，精子组蛋白高度乙酰化，表明它们的相关基因可能参与了胚胎的早期基因表达。
方法：使用原位杂交和免疫细胞化学技术，分析人（ n = 22）和mar（n = 6）睾丸和射精。
结果：在精子和圆形精子中都存在鱼精蛋白1和鱼精蛋白2的mRNA。所有针对乙酰化组蛋白的抗体在这些细胞中均显示阳性信号。人精原细胞对H4K8ac和H4K16ac呈阳性信号，而mar猴精原细胞对H4K8ac和H4K12ac呈阳性信号。在人类中，H4K16ac从生精上皮细胞周期的第III阶段开始就已经显示出与粗精子细胞的阳性免疫反应。所有抗体在两个物种的射精精子中均显示阳性免疫染色。
结论：普通mar猴是研究人类精子发生过程中核蛋白表达的合适动物模型。这两个物种的生精上皮组织相似，在圆形和延伸的精子细胞中，鱼精蛋白-1和鱼精蛋白2 mRNA的表达模式相同。睾丸精子和射精的精子中特异乙酰化组蛋白H4的存在表明这两种物种中鱼精蛋白交换的组蛋白不完全。由于已知乙酰化的组蛋白与参与基因表达的基因有关，因此普通mar猴将来可能被用作研究早期胚胎发育的模型，而在人类中，出于伦理原因，这是不可能的。

BACKGROUND & AIMS: :Exposure of male rats to cyclophosphamide, a commonly used anticancer and immunosuppressive drug, has been shown to alter fertility and progeny outcome in a male germ cell phase-specific manner. The effect of toxicant exposure on male germ cells depends in part on the stress response mechanisms present during the different stages of spermatogenesis. To assess how acute cyclophosphamide exposure affects the expression of stress response genes, we examined the expression of 216 genes, using gene expression arrays, in isolated rat spermatogenic cell types (pachytene spermatocytes, round spermatids, and elongating spermatids). Cyclophosphamide exposure affected gene expression in all cell types but most dramatically in round spermatids. Increased transcript levels were observed for 30 genes in round spermatids compared to seven genes in pachytene spermatocytes and two in elongating spermatids. The expression of genes involved in apoptosis, DNA-damage recognition and repair, transcriptional activation, and in the heat shock protein-chaperone response was most affected by cyclophosphamide in round spermatids. Our results demonstrate that cyclophosphamide alters the expression of stress response genes during spermatogenesis in a germ cell-specific manner. The greater response of round spermatids to cyclophosphamide suggests that this cell type may be more susceptible to the damaging effects induced by this drug, possibly due to the chromatin remodeling that is taking place at this stage of spermatogenesis. This observation is consistent with the reported higher level of abnormal progeny outcome seen when the germ cells were first exposed to cyclophosphamide as round spermatids.

背景与目标： ：雄性大鼠接触环磷酰胺是一种常用的抗癌和免疫抑制药物，已显示出以雄性生殖细胞阶段特异性的方式改变生育力和后代结局。有毒物质对雄性生殖细胞的影响部分取决于在精子发生不同阶段存在的应激反应机制。为了评估急性环磷酰胺暴露如何影响应激反应基因的表达，我们使用基因表达阵列检查了分离的大鼠生精细胞类型（粗大精细胞，圆形精细胞和伸长精细胞）中216个基因的表达。环磷酰胺暴露会影响所有细胞类型中的基因表达，但在圆形精子细胞中最显着。圆形精子中的30个基因的转录水平增加，而粗线精子细胞中的7个基因和伸长精子中的2个基因的转录水平增加。圆形精子中环磷酰胺对细胞凋亡，DNA损伤识别和修复，转录激活以及热休克蛋白-伴侣应答中涉及的基因表达的影响最大。我们的结果表明，环磷酰胺以生殖细胞特异性方式改变了精子发生过程中应激反应基因的表达。圆形精子对环磷酰胺的更大反应表明，这种细胞类型可能更容易受到这种药物诱导的破坏作用的影响，这可能是由于在精子发生这一阶段发生的染色质重塑。该观察结果与报告的较高水平的异常子代结局水平相一致，这是当生殖细胞首次以圆形精子细胞暴露于环磷酰胺时看到的。

BACKGROUND & AIMS: :The Azoospermia Factor c (AZFc) region on the Y chromosome long arm is one of the least stable regions in the human genome. It consists almost entirely of very long repeats and is prone to rearrangement. Numerous structures at AZFc have been identified, and some of them have been reported to be associated with male infertility. We screened 580 Han Chinese in Taiwan for AZFc deletion and duplication using three PCR assays, and characterized the DAZ genes in selected subjects with additional Southern analyses. About 9.5% of our subjects have AZFc partial deletion, 2.8% have partial deletion followed by duplication, and 1.7% have partial duplication. The overall rearrangement frequencies vary significantly between different Y chromosome haplogroups (Yhgs), ranging from 2.9% in O3e to 100% in N and Q. All individuals in Yhg-N lack the sY1191 marker, but one out of three of them actually have four DAZ genes, indicating further duplication after the b2/b3 deletion. Our additional screening of 142 oligospermic men and 107 fertile controls found no significant difference in the frequencies of the gr/gr and the b2/b3 deletion. However, the frequency of AZFc partial duplication in the infertile group (7.0%) was significantly higher than that in the fertile control group (0.9%) and the general Taiwanese population (1.7%). Our results indicate that AZFc partial deletion and partial duplication are common polymorphisms in Han Chinese, and that the AZFc partial duplication, but not the AZFc partial deletion, is a risk factor for male infertility in the Taiwanese population.

背景与目标： ：Y染色体长臂上的无精子因子c（AZFc）区是人类基因组中最不稳定的区域之一。它几乎完全由很长的重复组成，容易重新排列。已经确定了AZFc的许多结构，并且据报道其中一些与男性不育有关。我们使用三种PCR分析方法筛选了580位中国台湾汉族人群中AZFc缺失和重复的情况，并通过额外的Southern分析对了选定受试者中的DAZ基因进行了表征。我们的受试者中约有9.5％的人有AZFc部分缺失，有2.8％的人部分缺失，然后重复，有1.7％的人有部分重复。在不同的Y染色体单倍组（Yhgs）之间，总体重排频率差异显着，范围从O3e的2.9％到N和Q的100％。Yhg-N中的所有个体都缺乏sY1191标记，但是其中三分之一实际上有四个DAZ基因，表明b2 / b3缺失后进一步重复。我们对142名少精症男性和107名受精对照男性进行了额外筛查，发现gr / gr和b2 / b3缺失的频率没有显着差异。然而，不育组中AZFc部分复制的频率（7.0％）显着高于可育对照组（0.9％）和台湾普通人群（1.7％）。我们的结果表明AZFc部分缺失和部分重复是汉族人的常见多态性，而AZFc部分重复而不是AZFc部分缺失是台湾人群男性不育的危险因素。

BACKGROUND & AIMS: :Exposure to bisphenol A (BPA) has been related to male reproductive disorders. Since this endocrine disruptor also displays genotoxic and epigenotoxic effects, it likely alters the spermatogenesis, a process in which both hormones and chromatin remodeling play crucial roles. The hypothesis of this work is that BPA impairs early embryo development by modifying the spermatic genetic and epigenetic information. Zebrafish males were exposed to 100 and 2000 μg/L BPA during early spermatogenesis and during the whole process. Genotoxic and epigenotoxic effects on spermatozoa (comet assay and immunocytochemistry) as well as progeny development (mortality, DNA repairing activity, apoptosis and epigenetic profile) were evaluated. Exposure to 100 µg/L BPA during mitosis slightly increased sperm chromatin fragmentation, enhancing DNA repairing activity in embryos. The rest of treatments promoted high levels of sperm DNA damage, triggering apoptosis in early embryo and severely impairing survival. Regarding epigenetics, histone acetylation (H3K9Ac and H3K27Ac) was similarly enhanced in spermatozoa and embryos from males exposed to all the treatments. Therefore, BPA male exposure jeopardizes embryonic survival and development due to the transmission of a paternal damaged genome and of a hyper-acetylated histone profile, both alterations depending on the dose of the toxicant and the temporal window of exposure.

背景与目标： ：双酚A（BPA）的暴露与男性生殖系统疾病有关。由于这种内分泌干扰物还具有遗传毒性和表观遗传毒性作用，因此它很可能改变了精子发生过程，在此过程中激素和染色质的重塑都起着至关重要的作用。这项工作的假设是，双酚A通过修饰精子遗传和表观遗传信息来损害早期胚胎发育。斑马鱼的雄性在早期生精过程中和整个过程中都暴露于100和2000μg/ L BPA。评估了对精子的遗传毒性和表观遗传毒性作用（彗星试验和免疫细胞化学）以及子代发育（死亡率，DNA修复活性，细胞凋亡和表观遗传特征）。有丝分裂期间暴露于100μg/ L BPA会稍微增加精子染色质的碎片，从而增强胚胎中的DNA修复活性。其余的治疗促进了高水平的精子DNA损伤，触发了早期胚胎中的细胞凋亡，并严重损害了存活率。关于表观遗传学，在接受所有治疗的雄性精子和胚胎中，组蛋白乙酰化（H3K9Ac和H3K27Ac）同样得到增强。因此，由于父本受损基因组和高乙酰化组蛋白谱的传递，BPA男性暴露危害胚胎存活和发育，这两种改变均取决于毒物的剂量和暴露的时间窗。

BACKGROUND & AIMS: STUDY QUESTION:What is the consequence of Tex19.1 gene deletion in mice? SUMMARY ANSWER:The Tex19.1 gene is important in spermatogenesis and placenta-supported development. WHAT IS KNOWN ALREADY:Tex19.1 is expressed in embryonic stem (ES) cells, primordial germ cells (PGCs), placenta and adult gonads. Its invalidation in mice leads to a variable impairment in spermatogenesis and reduction of perinatal survival. STUDY DESIGN, SIZE, DURATION:We generated knock-out mice and ES cells and compared them with wild-type counterparts. The phenotype of the Tex19.1 knock-out mouse line was investigated during embryogenesis, fetal development and placentation as well as during adulthood. PARTICIPANTS/MATERIALS, SETTING, METHODS:We used a mouse model system to generate a mutant mouse line in which the Tex19.1 gene was deleted in the germline. We performed an extensive analysis of Tex19.1-deficient ES cells and assessed their in vivo differentiation potential by generating chimeric mice after injection of the ES cells into wild-type blastocysts. For mutant animals, a morphological characterization was performed for testes and ovaries and placenta. Finally, we characterized semen parameters of mutant animals and performed real-time RT-PCR for expression levels of retrotransposons in mutant testes and ES cells. MAIN RESULTS AND THE ROLE OF CHANCE:While Tex19.1 is not essential in ES cells, our study points out that it is important for spermatogenesis and for placenta-supported development. Furthermore, we observed an overexpression of the class II LTR-retrotransposon MMERVK10C in Tex19.1-deficient ES cells and testes. LIMITATIONS, REASONS FOR CAUTION:The Tex19.1 knock-out phenotype is variable with testis morphology ranging from severely altered (in sterile males) to almost indistinguishable compared with the control counterparts (in fertile males). This variability in the testis phenotype subsequently hampered the molecular analysis of mutant testes. Furthermore, these results were obtained in the mouse, which has a second isoform (i.e. Tex19.2), while other mammals possess only one Tex19 (e.g. in humans). WIDER IMPLICATIONS OF THE FINDINGS:The fact that one gene has a role in both placentation and spermatogenesis might open new ways of studying human pathologies that might link male fertility impairment and placenta-related pregnancy disorders. STUDY FUNDING/COMPETING INTEREST(S):This work was supported by the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé et de la Recherche Médicale (INSERM) (Grant Avenir), the Ministère de l'Education Nationale, de l'Enseignement Supérieur et de la Recherche, the Université de Strasbourg, the Association Française contre les Myopathies (AFM) and the Fondation pour la Recherche Médicale (FRM) and Hôpitaux Universitaires de Strasbourg.The authors have nothing to disclose.

背景与目标： 研究问题：小鼠Tex19.1基因缺失的后果是什么？
总结答案：Tex19.1基因在精子发生和胎盘支持的发育中很重要。
Tex19.1已在胚胎干（ES）细胞，原始生殖细胞（PGC），胎盘和成年性腺中表达。其在小鼠中的无效导致精子发生的可变性损伤和围产期生存期的减少。
研究设计，大小和持续时间：我们产生了敲除小鼠和ES细胞，并将其与野生型小鼠进行了比较。 Tex19.1基因敲除小鼠系的表型在胚胎发生，胎儿发育和胎盘形成以及成年期间进行了调查。
参与者/材料，设置，方法：我们使用小鼠模型系统生成了一个突变小鼠系，其中在种系中删除了Tex19.1基因。我们对Tex19.1缺陷型ES细胞进行了广泛的分析，并在将ES细胞注射入野生型胚泡后生成了嵌合小鼠，从而评估了它们的体内分化潜能。对于突变动物，对睾丸，卵巢和胎盘进行了形态学表征。最后，我们表征了突变动物的精液参数，并对突变睾丸和ES细胞中反转录转座子的表达水平进行了实时RT-PCR。
主要结果和可能的作用：虽然Tex19.1在ES细胞中不是必需的，但我们的研究指出，它对精子发生和胎盘支持的发育很重要。此外，我们观察到在Tex19.1缺陷的ES细胞和睾丸中II类LTR-反转录转座子MMERVK10C的过表达。
局限性，引起注意的原因：Tex19.1基因敲除表型的睾丸形态可变，从严重改变（在不育雄性中）到与对照组相比（在可育雄性中）几乎无法区分。睾丸表型的这种可变性随后阻碍了突变睾丸的分子分析。此外，这些结果是在具有第二种亚型（即Tex19.2）的小鼠中获得的，而其他哺乳动物仅具有一个Tex19（例如在人类中）。
结果的更广泛含义：一个基因在胎盘发育和精子发生中均起作用的事实可能会为研究可能与男性生育力障碍和胎盘相关的妊娠疾病相关的人类病理学开辟新途径。
研究资金/竞争兴趣：这项工作得到了国家科学研究所中心（CNRS），国家卫生研究中心（INSERM）（格兰特·阿韦尼尔），教育部的支持。法国国家安全研究与研究中心，斯特拉斯堡大学，法国精神病防治协会（AFM）和法国医学研究基金会（FRM）和斯特拉斯堡大学图书馆。

BACKGROUND & AIMS: :The Wnt/β-catenin pathway participates in many important physiological events such as cell proliferation and differentiation in the male reproductive system. We found that Kinesin-2 motor KIF3A is highly expressed during spermatogenesis in Eriocheir sinensis; it may potentially promote the intracellular transport of cargoes in this process. However, only a few studies have focused on the relationship between KIF3A and the Wnt/β-catenin pathway in the male reproductive system of decapod crustaceans. In this study, we cloned and characterized the CDS of β-catenin in E. sinensis for the first time. Fluorescence in situ hybridization and immunofluorescence results showed the colocalization of Es-KIF3A and Es-β-catenin at the mRNA and the protein level respectively. To further explore the regulatory function of Es-KIF3A to the Wnt/β-catenin pathway, the es-kif3a was knocked down by double-stranded RNA (dsRNA) in vivo and in primary cultured cells in testes of E. sinensis. Results showed that the expression of es-β-catenin and es-dvl were decreased in the es-kif3a knockdown group. The protein expression level of Es-β-catenin was also reduced and the location of Es-β-catenin was changed from nucleus to cytoplasm in the late stage of spermatogenesis when es-kif3a was knocked down. Besides, the co-IP result demonstrated that Es-KIF3A could bind with Es-β-catenin. In summary, this study indicates that Es-KIF3A can positively regulate the Wnt/β-catenin pathway during spermatogenesis and Es-KIF3A can bind with Es-β-catenin to facilitate the nuclear translocation of Es-β-catenin.

背景与目标： Wnt /β-catenin途径参与许多重要的生理事件，例如男性生殖系统中的细胞增殖和分化。我们发现Kinosin-2电机KIF3A在中华绒螯蟹的精子发生过程中高度表达。它可能会在此过程中促进货物的细胞内运输。然而，只有很少的研究集中在十足纲甲壳纲雄性生殖系统中KIF3A与Wnt /β-catenin途径之间的关系上。在这项研究中，我们首次克隆和表征了中华绒螯蟹中β-catenin的CDS。荧光原位杂交和免疫荧光结果显示Es-KIF3A和Es-β-catenin在mRNA和蛋白水平上共定位。为了进一步探索Es-KIF3A对Wnt /β-catenin途径的调控功能，在体内和中华绒螯蟹的原代培养细胞中，通过双链RNA（dsRNA）敲除了es-kif3a。结果表明，在es-kif3a敲除组中，es-β-catenin和es-dvl的表达降低。当es-kif3a被击倒时，精子形成后期Es-β-catenin的蛋白表达水平也降低，Es-β-catenin的位置从细胞核改变为细胞质。另外，co-IP结果表明Es-KIF3A可以与Es-β-catenin结合。总之，这项研究表明，Es-KIF3A可以在精子发生过程中正调控Wnt /β-catenin途径，Es-KIF3A可以与Es-β-catenin结合，促进Es-β-catenin的核转运。

BACKGROUND & AIMS: :Formin is one of the two major classes of actin binding proteins (ABPs) with nucleation and polymerization activity. However, despite advances in our understanding of its biochemical activity, whether and how formins generate specific architecture of the actin cytoskeleton and function in a physiological context in vivo remain largely obscure. It is also unknown how actin filaments generated by formins interact with other ABPs in the cell. Here, we combine genetic manipulation of formins mammalian diaphanous homolog1 (mDia1) and 3 (mDia3) with superresolution microscopy and single-molecule imaging, and show that the formins mDia1 and mDia3 are dominantly expressed in Sertoli cells of mouse seminiferous tubule and together generate a highly dynamic cortical filamentous actin (F-actin) meshwork that is continuous with the contractile actomyosin bundles. Loss of mDia1/3 impaired these F-actin architectures, induced ectopic noncontractile espin1-containing F-actin bundles, and disrupted Sertoli cell-germ cell interaction, resulting in impaired spermatogenesis. These results together demonstrate the previously unsuspected mDia-dependent regulatory mechanism of cortical F-actin that is indispensable for mammalian sperm development and male fertility.

背景与目标： ：Formin是具有成核和聚合活性的肌动蛋白结合蛋白（ABP）的两大类之一。然而，尽管我们对其生化活性的理解有了进步，但是在体内的生理情况下，福尔马林是否以及如何产生肌动蛋白细胞骨架的特定结构和功能仍不清楚。还不知道由福尔马林产生的肌动蛋白丝如何与细胞中的其他ABP相互作用。在这里，我们结合了formins哺乳动物透明的同系物1（mDia1）和3（mDia3）的基因操作与超分辨率显微镜和单分子成像，并显示formins mDia1和mDia3在小鼠曲细精管的Sertoli细胞中显性表达，并共同产生高度动态的皮质丝状肌动蛋白（F-actin）网状结构，与收缩性肌动蛋白束连续。 mDia1 / 3的丧失会破坏这些F-肌动蛋白的结构，诱导异位的非收缩性含espin1的F-肌动蛋白束，并破坏Sertoli细胞与生殖细胞的相互作用，从而导致精子发生受损。这些结果共同证明了皮质F-肌动蛋白的先前未曾怀疑的依赖mDia的调节机制，这对于哺乳动物的精子发育和雄性育性是必不可少的。

BACKGROUND & AIMS: -2

背景与目标： -2

BACKGROUND & AIMS: :Spermatogenesis is a process driven by stem cell, where germ cell cycle is under the control of a specific genotype species. Considering that Jundiá (Rhamdia quelen) is a Neotropical catfish with great economical importance and useful experimental model, little information is available on basic aspects of its reproductive biology, especially on spermatogenesis. As a result, this study aimed to characterize the male germ cells, estimate the duration of spermatogenesis and evaluate the expression of selected stem cell genes in Jundiá testis. Similar to other fish species, our results showed a remarkable decrease of germ cell nuclear volume during Jundiá spermatogenesis, particularly from type A undifferentiated to late type B spermatogonia and from diplotene to late spermatids. Using a S-phase marker, bromodeoxyuridine (BrdU), the combined duration of meiotic and spermiogenic phases in this species was estimated in approximately 7 days. This is considered very short when compared to mammals, where spermatogenesis last from 30 to 74 days. Selected stem cell genes were partially sequenced and characterized in Jundiá testis. Expression analysis showed higher plzf and pou5f3 mRNA levels in the cell fractions enriched by type A undifferentiated spermatogonia. These results were further confirmed by in situ hybridization that showed strong signal of plzf and pou5f3 mRNA in type A undifferentiated spermatogonia. Altogether, these information will expand our knowledge of the reproductive biology of this species, contributing to improve its production and management, and also for biotechnological applications, such as germ cell transplantation.

背景与目标： ：精子发生是由干细胞驱动的过程，其中生殖细胞的周期受特定基因型物种的控制。考虑到Jundiá（Rhamdia quelen）是一种新热带cat鱼，具有重要的经济意义和有用的实验模型，关于其生殖生物学的基本方面（尤其是精子发生）的信息很少。因此，本研究旨在表征雄性生殖细胞，估计精子发生的持续时间，并评估纯睾丸中所选干细胞基因的表达。与其他鱼类相似，我们的研究结果表明，在Jundiá精子发生过程中，生殖细胞核的数量显着减少，特别是从未分化的A型精子到晚期的B型精原细胞，从双倍戊烯到晚期的精子。使用S-阶段标记物，溴脱氧尿苷（BrdU），估计该物种中减数分裂和生精阶段的合并持续时间约为7天。与精子发生持续30至74天的哺乳动物相比，这被认为非常短。选定的干细胞基因在Jundiá睾丸中进行了部分测序和鉴定。表达分析显示，在未分化的A型精原细胞富集的细胞级分中，较高的plzf和pou5f3 mRNA水平较高。这些结果通过原位杂交进一步证实，该原位杂交显示在A型未分化精原细胞中plzf和pou5f3 mRNA的强信号。总而言之，这些信息将扩大我们对该物种生殖生物学的知识，有助于改善其生产和管理，并有助于生物技术应用，例如生殖细胞移植。

BACKGROUND & AIMS: RATIONALE:For the carriers of chromosome reciprocal translocation, the reason why some are fertile and others are infertile remains unclear. Here, we describe 2 patients who are carriers of chromosome 1q21 translocation with azoospermia. PATIENT CONCERNS:A 29-year-old male and a 33-year-old male presented at the clinic with a diagnosis of infertility. DIAGNOSIS:Both patients with azoospermia were diagnosed with Routine semen analysis, cytogenetic diagnosis and detection of serum reproductive hormones. The karyotype results of 2 patients were 46,XY,t(1;17)(q21;q23) and 46,XY,t(1;10)(q21;p12), respectively. INTERVENTIONS:After genetic counseling and informed consent, 1 patient (Case 2) chose microsopic testicular sperm extraction (micro-TESE). OUTCOMES:After micro-TESE, no sperm was found for the patient. Finally, both patients chose clinical treatment through artificial insemination with donor sperm. LESSONS:These outcomes suggest that breakpoint at 1q21 should be paid attention by physician in genetic counseling, may harbor some genes associated with spermatogenesis, and deserves further be studied on the function of related genes.

背景与目标： 理由：对于染色体易位的携带者，尚不清楚其中一些是可育的而另一些是不育的。在这里，我们描述了2名患者，它们是染色体1q21易位与无精子症的携带者。
患者注意事项：一名29岁的男性和33岁的男性在诊所被诊断出不育症。
诊断：两名无精症患者均经常规精液分析，细胞遗传学诊断和血清生殖激素检测。 2例患者的核型检查结果分别为46，XY，t（1; 17）（q21; q23）和46，XY，t（1; 10）（q21; p12）。
干预措施：经过遗传咨询和知情同意后，有1名患者（病例2）选择了显微睾丸精子摘除术（micro-TESE）。
结果：进行微TESE后，未发现患者的精子。最后，两名患者都选择了通过供体精子的人工授精进行临床治疗。
经验教训：这些结果表明，在基因咨询中应注意医师在1q21时的断点，其中可能包含一些与精子发生有关的基因，值得对相关基因的功能进行进一步研究。

BACKGROUND & AIMS: :The classification of azoospermia into obstructive or non-obstructive is largely based on medical history, physical examination and biochemical markers in serum and semen. However, the most accurate parameter for diagnosis is the testicular histology. The predictive value of the percutaneous epididymal sperm aspiration (PESA), FSH, LH, testosterone, inhibin-B and testicular volume was investigated for their accuracy to predict a complete spermatogenesis (Johnsen score > or =8) in order to replace the testicular histology. The specificity and sensitivity of FSH, inhibin-B, LH, testosterone, testicular volume, and the presence of sperm in a PESA procedure was evaluated in 147 azoospermic males attending the centre for infertility diagnosis. A positive PESA outcome presented the highest sensitivity and specificity to predict a Johnsen score > or =8 (93 and 94% respectively) compared with FSH (90 and 19%), inhibin-B (88 and 57%) and testicular volume (95 and 45%). Differences in clinical presentation were observed between patients with positive sperm retrieval with PESA, depending on the aetiology of obstruction. In conclusion, the presence of spermatozoa in the epididymis (PESA+) correlates with a Johnsen score > or =8 and is the most accurate parameter to predict complete spermatogenesis compared with clinical or biochemical parameters. Between obstructive azoospermic patients, the clinical parameters observed varied according to the aetiology.

背景与目标： ：无精子症分为阻塞性或非阻塞性的主要依据是病史，体格检查以及血清和精液中的生化指标。但是，最准确的诊断参数是睾丸组织学。研究了经皮附睾精子抽吸术（PESA），FSH，LH，睾丸激素，抑制素B和睾丸体积的预测价值，以预测其完整精子发生的准确性（Johnsen评分>或= 8），以替代睾丸组织学。 。在147名参加不育症诊断中心的无精子症男性中，评估了PESH程序中FSH，抑制素-B，LH，睾丸激素，睾丸体积和精子的存在的特异性和敏感性。与FSH（90％和19％），抑制素B（88％和57％）和睾丸体积（95）相比，PESA阳性结果预测Johnsen评分>或= 8的敏感性和特异性最高（分别为93％和94％），B抑制素（88％和57％）。和45％）。根据梗阻的病因，在PESA精子检出阳性的患者之间观察到临床表现的差异。总之，附睾中的精子（PESA）的存在与Johnsen评分>或= 8相关，并且与临床或生化指标相比，它是预测完全精子发生的最准确参数。在阻塞性无精子症患者之间，观察到的临床参数根据病因而有所不同。

BACKGROUND & AIMS: :Graphene oxide (GO) has the potential for wide applications, which necessitates an intensive investigation of its potential hazard on human and environmental health. Even if previous studies show reproductive toxicity in the nematode Caenorhabditis elegans, the mechanisms of reproductive toxicity by GO are poorly understood. To understand the underlying mechanisms of GO-induced reproductive toxicity, we investigated the interaction between GO and C. elegans using Raman spectroscopy, sperm counts produced by spermatogenesis, progeny and analyzed the fatty acid metabolism using molecular techniques. GO-characteristic Raman spectral bands measured throughout C. elegans, brood size and Hoecst staining of dissected gonads clearly showed GO accumulation in the reproductive organs, reduced progeny and low sperm counts, which are possibly direct results of the reproductive toxicity from GO exposure. Interestingly, reduced fatty acid metabolites, such as stearic, oleic, palmitoleic, and palmitic acids, were found with GO exposure. We found that GO increased intestinal fat accumulation in wild type N2, fat-5(tm420), and fat-7(wa36) mutants, whereas it decreased fat storage in the fat-6(tm331) and nhr-49(nr2041) mutants. GO exposure affected C. elegans fat accumulation and consumption, which was possibly regulated by daf-16 and nhr-80 gene activity. Also, GO exposure suppressed the survival of long-lived fat-5(tm420) mutants, whereas it increased the survival of short-lived nhr-49(nr2041) mutants. Hence, our studies collectively indicated that GO accumulation in reproductive organs, suppression of spermatogenesis, and the alteration of fatty acid metabolism play critical roles in understanding mechanisms of toxicity in C. elegans.

背景与目标： ：氧化石墨烯（GO）具有广泛的应用潜力，因此有必要深入研究其对人体和环境健康的潜在危害。即使先前的研究表明线虫秀丽隐杆线虫的生殖毒性，对GO生殖毒性的机制仍知之甚少。为了了解GO诱导的生殖毒性的潜在机制，我们使用拉曼光谱法研究了GO和秀丽隐杆线虫之间的相互作用，由精子发生产生的精子数量，子代，并使用分子技术分析了脂肪酸代谢。在整个秀丽隐杆线虫中测得的GO特征拉曼光谱带，切出的性腺的亲代大小和Hoecst染色清楚地表明GO在生殖器官中积累，子代减少和精子数量减少，这可能是GO接触导致生殖毒性的直接结果。有趣的是，GO暴露后发现脂肪酸的代谢产物减少，例如硬脂酸，油酸，棕榈油酸和棕榈酸。我们发现GO增加了野生型N2，fat-5（tm420）和fat-7（wa36）突变体中肠道脂肪的积累，而它降低了fat-6（tm331）和nhr-49（nr2041）突变体中的脂肪储存。 GO暴露影响线虫脂肪的积累和消耗，这可能受daf-16和nhr-80基因活性的调节。此外，GO暴露抑制了长寿命的fat-5（tm420）突变体的存活，而增加了短寿命的nhr-49（nr2041）突变体的存活。因此，我们的研究共同表明，GO在生殖器官中的蓄积，精子生成的抑制以及脂肪酸代谢的改变在了解秀丽隐杆线虫的毒性机理中起着至关重要的作用。

BACKGROUND & AIMS: OBJECTIVE:To assess the treatment outcome after varicocele repair in azoospermic men and to correlate this outcome with the testicular histology patterns. DESIGN:Prospective study. SETTING:Academic medical centers. PATIENT(S):Medical records of 27 azoospermic men, who underwent testis biopsy and microsurgical repair of clinical varicocele between July 1999 and May 2004, were reviewed. INTERVENTION(S):Twenty-seven azoospermic men underwent testis biopsy and microsurgical repair of clinical varicocele. All patients had at least two semen analyses showing azoospermia taken before the surgery and two semen analyses postoperatively. MAIN OUTCOME MEASURE(S):Semen analysis after varicocelectomy. RESULT(S):Hypospermatogenesis was identified in 9, maturation arrest in 8, and germ cell aplasia in 10 men. Induction of spermatogenesis was achieved in nine men (33.3%). Of these, four had germ cell aplasia, three had maturation arrest, and two had hypospermatogenesis. The improvement in sperm concentration and motility ranged from 1.2 x 10(6)/mL to 8.9 x 10(6)/mL, and from 24% to 75.7%, respectively. Of these nine patients with improvement in semen quality, five relapsed into azoospermia 6 months after the recovery of spermatogenesis (four germ cell aplasia and one maturation arrest). One patient with maturation arrest established pregnancy. CONCLUSION(S):Azoospermic patients may have an improvement in semen quality after varicocelectomy. Semen samples may be cryopreserved after an initial improvement after varicocelectomy.

背景与目标： 目的：评估无精症男性精索静脉曲张修复后的治疗效果，并将其与睾丸组织学模式相关联。
设计：前瞻性研究。
地点：学术医疗中心。
病人：回顾了1999年7月至2004年5月间接受睾丸活检和临床精索静脉曲张显微手术修复的27例无精子症患者的医学记录。
干预：27名无精子症男子接受了睾丸活检和临床精索静脉曲张的显微外科手术修复。所有患者在手术前至少进行了两次精液分析，显示出无精子症，术后进行了两次精液分析。
主要观察指标：精索静脉曲张切除术后的精液分析。
结果：在9例中发现了过度生精，在8例中发现了成熟停止，在10例男性中发现了生殖细胞发育不良。诱导了9名男性（33.3％）的精子发生。其中，四个患有生殖细胞发育不全，三个具有成熟停滞，两个具有低精子发生。精子浓度和运动力的改善范围分别为1.2 x 10（6）/ mL至8.9 x 10（6）/ mL，以及24％至75.7％。在这9例精液质量改善的患者中，有5例在精子发生恢复6个月后复发至无精子症（4例生殖细胞发育不良和1例成熟停止）。一名成熟逮捕患者确定怀孕。
结论：精索静脉曲张切除术后无精症患者的精液质量可能有所改善。精索静脉曲张切除术后初步改善后，可以冷冻保存精液样本。