Assessment of ameliorative effects of Moringa oleifera Lam. on epididymal dysfunctions and fertility in streptozotocin-induced diabetic rats

Objective:To assess the protective effects of 70%ethanolic extract of Moringa(M.)oleifera leaves on fertility and epididymal dysfunctions in diabetic rats.Methods:Thirty-six male Wistar rats were divided into six groups,each with six rats.GroupⅠwas the normal control group receiving vehicle(0.5 mL of distilled water/rat/day)orally;group Ⅱ,Ⅲ,Ⅳ,Ⅴ and Ⅵ were injected intraperitoneally with 60 mg/kg of streptozotocin once to induce type Ⅰ diabetes.Then,the diabetic rats in group Ⅱ receiving vehicle(0.5 mL of distilled water/rat/day)orally;the diabetic rats in groups Ⅲ,Ⅳ and Ⅴ were orally treated with M.oleifera leaf extract at dose of 100,250,and 500 mg/kg body weight(b.wt.)/day,respectively;the diabetic rats in group Ⅵ were administrated with reference drug glibenclamide(5 mg/kg b.wt./day).The treatment lasted for 60 days.Sperm parameters(sperm count,motility and viability),fertility index,litter size,lipid peroxidation and antioxidative defense markers(i.e.superoxide dismutase,catalase,glutathione,and ascorbic acid)in the epididymal tissue were determined.Histopathological changes in epididymis were also evaluated.Results:Administration of different doses of M.oleifera leaf extract in diabetic rats induced a remarkable dose dependent improvement in sperm parameters,fertility index,litter size,epididymal antioxidant status and also restored histopathological changes as compared to the diabetic control group.These findings were comparable with reference drug.Conclusions:M.oleifera leaf extract possesses significant antioxidant activities as well as beneficial effects on epididymal dysfunctions in diabetic male rats.

Knockdown of HE4 suppresses tumor growth and invasiveness in lung adenocarcinoma through regulation of EGFR signaling

It has been shown that the high expression of human epididymis protein 4(HE4)in most lung cancers is related to the poor prognosis of patients,but the mechanism of pathological transformation of HE4 in lung cancer is still unclear.The current study is expected to clarify the function and mechanism of HE4 in the occurrence and metastasis of lung adenocarcinoma(LUAD).Immunoblotting evaluated HE4 expression in lung cancer cell lines and biopsies,and through analysis of The Cancer Genome Atlas(TCGA)dataset.Frequent HE4 overexpression was demonstrated in LUAD,but not in lung squamous cell carcinoma(LUSC),indicating that HE4 can serve as a biomarker to distinguish between LUAD and LUSC.HE4 knockdown significantly inhibited cell growth,colony formation,wound healing,and invasion,and blocked the G1-phase of the cell cycle in LUAD cell lines through inactivation of the EGFR signaling downstream including PI3K/AKT/mTOR and RAF/MAPK pathways.The first-line EGFR inhibitor gefitinib and HE4 shRNA had no synergistic inhibitory effect on the growth of lung adenocarcinoma cells,while the third-line EGFR inhibitor osimertinib showed additive anti-proliferative effects.Moreover,we provided evidence that HE4 regulated EGFR expression by transcription regulation and protein interaction in LUAD.Our findings suggest that HE4 positively modulates the EGFR signaling pathway to promote growth and invasiveness in LUAD and highlight that targeting HE4 could be a novel strategy for LUAD treatment.

HE4与慢性心力衰竭患者心功能的相关性

目的探讨慢性心力衰竭患者HE4水平与心功能的关系。方法选取因慢性心力衰竭住院的200例患者为研究对象,平均年龄(71.81±12.49)岁。将心功能Ⅰ-Ⅱ级定义为心功能不全低危组,将心功能Ⅲ-Ⅳ级定义为心功能不全高危组。多因素Logistic回归分析评估导致慢性心力衰竭发生的危险因素。结果 BMI、HDL-C等指标心功能不全高危组与心功能不全低危组组间差异无统计学意义(P≥0.05);而LDL-C、CRP、HE4、UA等指标两组间差异有统计学意义(P<0.05)。当调节变量包括年龄、性别、HE4以及NT-pro BNP时,HE4和NT-pro BNP是与慢性心力衰竭患者心功能密切相关的危险因素。而当调节变量包括年龄、性别、HE4、NT-pro BNP、吸烟、饮酒、HDL-C、LDL-C以及UA时,结果提示性别、HE4和NT-pro BNP是与慢性心力衰竭患者心功能密切相关的危险因素。结论性别、HE4和NT-pro BNP是与慢性心力衰竭患者心功能密切相关的危险因素,慢性心力衰竭患者HE4水平与心功能的分级存在相关关系。

人附睾蛋白4与溶血磷脂酸联合检测对子宫内膜癌的临床诊断价值分析

目的探讨血清人附睾蛋白4(HE4)和溶血磷脂酸(LPA)联合检测对子宫内膜癌的诊断价值。方法采用酶联免疫吸附法和电化学发光法分别检测子宫内膜癌患者、子宫内膜异位症患者、功能失调性子宫出血患者及健康者血清HE4和血浆LPA水平,比较各研究组HE4和LPA水平及阳性率,分析HE4、LPA单项及联合检测对子宫内膜癌的诊断效能。结果子宫内膜癌组HE4、LPA水平较其他3组明显升高(P<0.05);HE4单项检测诊断子宫内膜癌的灵敏度为48.58%、特异度为95.74%,LPA单项检测的诊断灵敏度为68.96%、特异度为87.23%,二者联合检测的诊断灵敏度为75.86%、特异度为87.23%,联合检测的诊断灵敏度较LPA、HE4单项增高(P<0.05)。结论 HE4和LPA联合检测对子宫内膜癌的诊断效能优于二者单项检测。

人附睾蛋白4与癌抗原125在卵巢癌二元论模型中的应用

目的:探讨血清人附睾蛋白4(HE4)、CA125对上皮性卵巢癌(EOC)二元论模型的应用价值。方法:选取2012年2月—2013年4月间因盆腔包块或卵巢囊肿住院拟行手术的患者,包括妇科炎症65例,子宫内膜异位症(EMs)509例,卵巢良性肿瘤887例,卵巢交界性上皮瘤73例,EOC 132例(Ⅰ型78例,Ⅱ型54例),以及正常妇女71例。采用电化学发光法检测其血清HE4、CA125水平,并评估其应用价值。结果:EOC患者血清HE4和CA125水平与国际妇产科联盟(FIGO)分期呈正相关(rs分别为0.534,0.541,均P=0.000);各期Ⅱ型EOC HE4、CA125表达水平高于同期Ⅰ型EOC,高级浆液性癌高于低级浆液性癌,各期EOC HE4表达水平高于妇科良性疾患各组(P<0.05);Ⅰ期EOC患者血清CA125表达与妇科炎症、EMs组差异无统计学意义(均P>0.05),Ⅱ、Ⅲ期Ⅰ型EOC患者血清CA125表达与EMs组差异无统计学意义(均P>0.05)。HE4、CA125检测EOC的受试者工作曲线下面积(ROC-AUC)分别为0.877,0.786;准确度为90.0%,70.5%;敏感度67.4%,69.7%;特异度92.1%,72.6%;约登指数0.595,0.403。结论:HE4单项检测EOC的特异度优于CA125,单独血清HE4及其与CA125联合检测对EOC早期诊断、分期、病理分型及预后评估都有一定意义,可以作为诊断EOC的重要指标。

人血清附睾蛋白4和糖类抗原125联合检测对卵巢癌的诊断价值

目的测定卵巢癌患者血清附睾蛋白4(HE4)和糖类抗原(CA)125水平,探讨血清HE4和CA125联合测定对卵巢癌的诊断价值。方法选择佛山市第一人民医院、佛山市第二人民医院、佛山中医院、佛山禅城区中心医院2012年1月至2013年5月收治的160例住院或门诊患者,将其分为卵巢癌组91例和卵巢良性肿瘤组69例。同时选择同期健康体检女性60例健康对照组。采用电化学发光免疫分析测定血清HE4和CA125水平。并检测卵巢癌病理分期与HE4、CA125水平的关系。结果卵巢癌患者血清HE4、CA125水平均明显高于卵巢良性肿瘤组和健康对照组(P<0.01);指标单独检测,HE4敏感性、特异性和诊断效率均高于CA125;并联试验时,敏感性明显提高,串联试验时,特异性明显提高。单独和联合检测中,并联试验对卵巢癌阳性检出率最高。卵巢癌患者随着病理分期升高血清HE4和CA125水平依次升高,但卵巢癌患者Ⅰ期血清CA125水平与卵巢良性肿瘤组比较,差异无统计学意义(P>0.05)。结论与CA125相比,HE4为最佳单项卵巢癌肿瘤标志物,联合检测并联试验可提高诊断敏感性,串联试验可提高诊断特异性。

Epididymosomes are involved in the acquisition of new sperm proteins during epididymal transit

During epididymal transit, spermatozoa acquire new proteins. Some of these newly acquired proteins behave as integral membrane proteins, including glycosylphosphatidylinositol （GPI）-anchored proteins. This suggests that the secreted epididymal proteins are transferred to spermatozoa by an unusual mechanism. Within the epididymal lumen, spermatozoa interact with small membranous vesicles named epididymosomes. Many proteins are associated with epididymosomes and the protein composition of these vesicles varies along the excurrent duct and differs from soluble intraluminal proteins. Some epididymosome-associated proteins have been identified and their functions in sperm maturation hypothesized. These include P25b, a zona pellucida binding protein, macrophage migration inhibitory factor, enzymes of the polyol pathway, HE5/CD52, type 5 glutathione peroxidase, and SPAM 1 or PH-20. The electrophoretic patterns of proteins associated to epididymosomes are complex and some of these proteins are transferred to defined surface domains of epididymal spermatozoa. Epididymosomes collected from different epididymal segments interact differently with spermatozoa. This protein transfer from epididymosomes to spermatozoa is timedependent, temperature-dependent and pH-dependent, and is more efficient in the presence of zinc. Some proteins are segregated to lipid raft domains of epididymosomes and are selectively transferred to raft domains of the sperm plasma membrane. Some evidence is presented showing that epididymosomes are secreted in an apocrine manner by the epididymal epithelial cells. In conclusion, epididymosomes are small membranous vesicles secreted in an apocrine manner in the intraluminal compartment of the epididymis and play a major role in the acquisition of new proteins by the maturing spermatozoa. （Asian J Androl 2007 July; 9： 483-491）

Effect of experimental varicocele on structure and function of epididymis in adolescent rats

<abstract>Aim: To study the effect of experimental left varicocele (ELV) on epididymal structure and function in adolescent Sprague-Dawley rats. Methods: ELV was induced by partial ligation of the left renal vein. Sham-operated animals served as the controls. Four and 8 weeks after the operation, the histological, ultrastructural and biochemical (alpha-glucosidase activity and carnitine content) changes in different segments of the epididymis were observed. Results: In the treated animals, there were degeneration of the epididymal epithelium and edema of the interstitial tissue; numerous shedding cells, residual bodies, deformed sperm and macrophages appeared in the epididymal lumen. Morphometric measurement indicated a significant reduction in the epididymal tubular diameter (P<0.05) and a significant increase in the epididymal interstitial area (P<0.05) compared with the controls. Ultrastructural study showed sparse microvilli of the columnar epithelium, increased and enlarged lysosomes in the principal cells with defected organelles and the presence of large cytoplasmic vacuoles. The protein and carnitine contents and the alpha-glucosidase activity in the caput, corpus and cauda epididymis of the ELV rats were lower than those of the controls (P< 0.05). Conclusion: There were structural and functional changes in the epididymis of adolescent ELV rats, which may contribute to the infertility caused by varicocele.

Diffusion-weighted MR imaging of normal and abnorma scrotum： preliminary results

Magnetic resonance （MR） imaging of the scrotum represents an important supplemental diagnostic tool in the evaluation of scrotal diseases. Diffusion-weighted （DW） MR imaging is a developing technique, proved to improve tissue characterization. We evaluated the feasibility and diagnostic performance of DW MR imaging in the detection and characterization of scrotal lesions. We retrospectively evaluated 31 scrotal lesions （23 intratesticular and 8 extratesticular） in 26 men. All MR examinations were performed on a 1.5-T unit, using a pelvic-phased array coil. DW sequences were obtained using a single shot, multislice spin echo planar diffusion pulse sequence and a b factor of 0 and 900 s mm-2. The DW MR characteristics and the apparent diffusion coefficient （ADC） values of normal scrotal contents and scrotal diseases were evaluated. Comparison between the ADC values of normal scrotum, benign lesions and scrotal malignancies was performed. The accuracy of conventional sequences, DW images alone and DW imaging combined with conventional images in differentiating benign from malignant scmtal lesions was calculated. The ADC values of testicular malignancies were different from those of normal testis and benign intratesticular lesions, and the ADC values of benign extratesticular lesions from those of normal epididymis （P〈0.05）. The overall accuracy of conventional imaging, DW imaging alone and DW MR combined with conventional sequences in the characterization of intratesticular lesions was 91%, 87% and 100%, respectively. Our findings suggest that DW MR imaging and ADC values may provide valuable information in the diagnosis and characterization of scrotal diseases.

A modified single-armed technique for microsurgical vasoepididymostomy

This study is to evaluate the effectiveness of a modified single-armed suture technique for microsurgical vasoepididymostomy （VE） in patients with epididymal obstructive azoospermia. From September 2011 to December 2011, microsurgical two-suture longitudinal intussusception VEs were performed using our modified single-armed suture technique in 17 men with epididymal obstructive azoospermia at our hospital. Two of these patients underwent repeated VEs after previous failed VEs, and one patient underwent unilateral VE because of an occlusion of the left abdominal vas deferens. The presence of sperm in the semen sample at 3 months postoperation was used as the preliminary endpoint of this study. Each patient provided at least one semen sample at the 3-month time point, and the patency was assessed by the reappearance of sperm （〉 104 m I- 1） in the semen. The mean operative time for the modified technique was 219 min. Patency was noted in 10 men （58.8%）, including one patient who underwent repeated VE. The patient who underwent unilateral anastomosis manifested no sperm postoperatively in his semen. Sperm granulomas were not detected in this cohort. The results of this study demonstrate that our modified technique for microsurgical longitudinal intussusception VE is effective. We believe that it is a practical alternative that may reduce operation time and obviate the suture crossing.

Participation of epididymal cysteine-rich secretory proteins in sperm-egg fusion and their potential use for male fertility regulation

Rat protein DE is an androgen-dependent cysteine-rich secretory protein （CRISP） synthesized by proximal epididymal regions. DE, also known as CRISP-1, is localized on the equatorial segment of acrosome-reacted spermatozoa and participates in gamete fusion through binding to egg complementary sites. Immunization of rats with DE inhibits fertility and sperm fusion ability, suggesting that DE represents a good epididymal contraceptive target. Recombinant DE fragments and synthetic peptides revealed that DE binds to the egg via a 12-amino acid region of an evolutionarily conserved motif, Signature 2 （S2）. The ability of other CRISP to bind to the rat egg was correlated with their S2 amino acid sequences. Although testicular protein Tpx- 1 （CRISP-2） was capable of binding to rodent eggs, human epididymal AEG-related protein （ARP） and helothermine （from lizard saliva） were not. The S2 region presented only two substitutions in Tpx-1 and four in ARP and helothermine, compared with the DE S2, suggesting that this amino acid sequence was relevant for egg interaction. Studies with Tpx- 1 and anti-Tpx- 1 revealed the participation of this protein in gamete fusion through binding to complementary sites in the egg. In competition studies, DE reduced binding of Tpx- 1 dose-dependently, indicating that both CRISP share the egg complementary sites. That anti-DE and anti-Tpx-1 inhibit sperm-egg fusion while recognizing only the corresponding proteins, suggests functional cooperation between these homologous CRISP to ensure fertilization success. These results increase our understanding of the molecular mechanisms of gamete fusion and contribute to the development of new and safer fertility regulating methods. （Asian J Androl 2007 July; 9： 528-532）

Proteomic changes in mammalian spermatozoa during epididymal maturation

Epididymal maturation is associated with the activation of a cAMP-induced tyrosine phosphorylation cascade, which is ultimately associated with the expression of capacitation-dependent sperm functions, such as hyperactivated movement and acrosomal exocytosis. As spermatozoa progress through the epididymis they first acquire the capacity to phosphorylate tyrosine on targets on the principal piece, followed by the midpiece. By the time these cells have reached the cauda epididymidis they can phosphorylate the entire tail from neck to endpiece. This particular pattern of phosphorylation is associated with the ontogeny of fully functional spermatozoa that are capable of fertilizing the oocyte. Proteomic analyses indicate that this change is associated with the phosphorylation of several mitochondrial proteins, creation of a mitochondrial membrane potential and activation of mitochondrial free radical generation. At least in rodent species, activation of sperm mitochondria appears to be a particularly important part of epididymal maturation. （Asian J Androl 2007 July; 9： 554-564）

Histological changes of the testis and epididymis in adult rats as a result of Leydig cell destruction after ethane dimethane sulfonate treatment： a morphometric study

Aim： To quantitatively study the histological changes of the testis and epididymis as a result of a drastic reduction of testosterone secretion. Methods： Fourteen adult Sprague-Dawley rats were injected intraperitoneally with ethane dimethane sulfonate （EDS, 75 mg/kg） and the same number of animals were injected with normal saline as a control. At days 7 and 12 （after treatment）, respectively, half of the animals from each group were killed. The testes and epididymides were removed and tissue blocks embedded in methacrylate resin. The cell number per testis was estimated using the stereological optical disector and some other parameters were obtained using other morphometric methods. Results： The EDS treatment resulted in an almost complete elimination of Leydig cells but had no effect on the numbers of Sertoli cells per testis. At day 7 after EDS treatment, many elongated spermatids were retained in the seminiferous epithelium and many round spermatids could be seen in the epididymal ducts. At day 12, a looser arrangement of spermatids and spermatocytes became evident, with apparent narrow empty spaces being formed between germ cells in an approximately radial direction towards the tubule lumen; the numbers （per testis） of non-type B spermatogonia and spermatocytes were similar to controls, whereas that of type B spermatogonia increased by 59%, and that of early round, elongating and late elongated spermatids decreased by 37%, 72% and 52%, respectively. Conclusion： The primary spermatogenic lesions following EDS administration were （i） spermiation failure and （ii） detachment of spermatids and spermatocytes associated with impairment in spermiogenesis and meiosis.

Effect of methoxychlor on antioxidant system of goat epididymal sperm in vitro

Aim: To evaluate the effect of methoxychlor on the antioxidant system of goat epididymal sperm. Methods:Epididymis of adult goat was obtained from local slaughter houses and sperm were collected by chopping the epididymisin modified Ringer's phosphate solution (RPS). After several washings, the sperm samples were dispersed in RPS andincubated with methoxychlor (1μmol/L, 10μmol/L and 100μmol/L) and methoxychlor + vitamin C (100μmol/Leach) for 3 h at 32℃. After incubation, the sperm motility and viability were assessed. An aliquot of sperm samplewas homogenized, centrifuged and used for the assay of superoxide dismutase, glutathione peroxidase, glutathione re-ductase and lipid peroxidation. Results; In methoxychlor-incubated sperm and in sperm co-incubated with methoxy-chlor and vitamin C, the sperm motility and viability showed no significant changes as compared to the correspondingcontrols. In methoxychlor-incubated sperm the activity of superoxide dismutase, glutathione reductase and glutathioneperoxidase were decreased while lipid peroxidation was increased in a dose-dependent manner. Co-incubation of spermwith methoxychlor and vitamin C showed no changes in the activity of superoxide dismutase, glutathione reductase andglutathione peroxidase and in the level of lipid peroxidation. Conclusion; Methoxychlor induced oxidative stress inepididymal sperm of goats by decreasing the levels of antioxidant enzymes. Co-incubation of sperm with methoxychlorand vitamin C, a natural antioxidant, reversed the effect of methoxychlor. (Asian J Androl 2001 Dec; 3; 285 - 288)

Effects of tripchlorolide on the epididymides and testes of rats

Aim: To further evaluate the antifertility effects of tripchorolide, a derivative of triptolide produced at the extraction pro-cedure of Tripterygium wilfordii Hook. f., in male rats and to investigate its sites and possible mechanisms of action.Methods: In male rats, tripchlorolide was given by oral garage at a dose of 50 ug.kg～(-l).d～(-1) for 5 weeks, fertility wasassessed by mating tests, and biochemical indices and light microscopic observation of the epididymides and testes werealso performed. Results: Administration of tripchlorolide at 50 ugg.kg～(-l)-d～(-1) for 3 weeks did not influence the fertilityof male rats, but 5-week treatment rendered the rats infertile. The density and motility of spermatozoa collected fromcauda epididymides were reduced significantly. The epididymal weights, as well as the L-carnitine concentration and α-glucosidase content in the epididymal fluid were decreasd. There were no significant differences in α-glucosidase andacid phosphatase (ACP) in caput epididymal homogenates between the control and the experimental rats. Obvious mor-phological changes were observed in the epididymal spermatozoa, mainly including head and tail separation or acrosomecurving. Sloughed spermatids were found in the seminifeous and epididymal tubules. In tesficular homogenates,tripchlorolide had no influence on the lactate dehydrogenase-C_4 (LDH-C_4) and hyaluronidase activities. No apparentlesions were observed in the seminiferous and epididymal epithelium. Conclusion: At the dose level employed,tripchlorolide has a significant effect on the fertility in male rats and the primary sites of action may be spermatids and tes-ticular and epididymal spermatozoa. (Asian J Androl 1999 Sep ; 1: 121 - 125)

Identification of epididymis-specific transcripts in the mouse and rat by transcriptional profiling

As part of our efforts to identify novel contraceptive targets in the epididymis we performed transcriptional profiling on each of the 10 and 19 segments of the mouse and rat epididymidis, respectively, using Affymetrix whole genome microarrays. A total of 17 096 and 16 360 probe sets representing transcripts were identified as being expressed in the segmented mouse and rat epididymal transcriptomes, respectively. Comparison of the expressed murine transcripts against a mouse transcriptional profiling database derived from 22 other mouse tissues identified 77 transcripts that were expressed uniquely in the epididymis. The expression of these genes was further evaluated by reverse transcription polymerase chain reaction （RT-PCR） analysis of RNA from 21 mouse tissues. RT-PCR analysis confirmed epididymis-specific expression of Defensin Beta 13 and identified two additional genes with expression restricted only to the epididymis and testis. Comparison of the 16 360 expressed transcripts in the rat epididymis with data of 21 other tissues from a rat transcriptional profiling database identified 110 transcripts specific for the epididymis. Sixty-two of these transcripts were further investigated by qPCR analysis. Only Defensin 22 （E3 epididymal protein） was shown to be completely specific for the epididymis. In addition, 14 transcripts showed more than 100-fold selective expression in the epididymis. The products of these genes might play important roles in epididymal and/or sperm function and further investigation and validation as contraceptive targets are warranted. The results of the studies described in this report are available at the Mammalian Reproductive Genetics （MRG） Database （http：//mrg. genetics.washington.edu/）. （Asian J Androl 2007July; 9： 522-527）

The role of cysteine-rich secretory proteins in male fertility

The cysteine-rich secretory proteins （CRISPs） are a subgroup of the CRISP, antigen 5 and Pr-1 （CAP） protein superfamily, and are found only in vertebrates. They show a strong expression bias to the mammalian male reproductive tract and the venom of poisonous reptiles. Within the male reproductive tract CRISPs have been implicated in many aspects of male germ cell biology spanning haploid germ cell development, epididymal maturation, capacitation, motility and the actual processes of fertilization. At a structural level, CRISPs are composed of two domains, a CAP domain, which has been implicated in cell-cell adhesion, and a CRISP domain, which has been shown to regulate several classes of ion channels across multiple species. Herein, we will review the current literature on the role of CRISPs in male fertility, and by inference to related non-mammalian protein, infer potential biochemical functions.

Stimulating effects of quercetin on sperm quality and reproductive organs in adult male rats

Aim： To investigate effects of quercetin on weight and histology of testis and accessory sex organs and on sperm quality in adult male rats. Methods： Male Sprague-Dawley rats were injected s.c. with quercetin at the dose of 0, 30, 90, or 270 mg/kg body weight/day （hereafter abbreviated Q0, Q30, Q90 and Q270, respectively）, and each dose was administered for treatment durations of 3, 7 and 14 days. Results： From our study, it was found that the effects of quercetin on reproductive organs and sperm quality depended on the dose and duration of treatment. After Q270 treatment for 14 days, the weights of testes, epididymis and vas deferens were significantly increased, whereas the weights of seminal vesicle and prostate gland were significantly decreased, compared with those of Q0. The histological alteration of those organs was observed after Q270 treatment for 7 days as well as 14 days. The sperm motility, viability and concentration were significantly increased after Q90 and Q270 injections after both of 7 and 14 days. Changes in sperm quality were earlier and greater than those in sex organ histology and weight, respectively. Conclusion： Overall results indicate that quercetin might indirectly affect sperm quality through the stimulation of the sex organs, both at the cellular and organ levels, depending on the dose and the duration of treatment. Therefore, the use of quercetin as an alternative drug for treatment of male infertility should be considered. （Asian J Androl 2008 Mar. 10： 249-258）

Quantitative (stereological) study on the spermatozoal storage capacity of epididymis in rats and monkeys

Aim: To investigate the spermatozoal production rate of the testis and the spermatozoal storage capacity of the epi-didymis in monkeys and rats. Methods: The number of the late spermatids (steps 13 - 14 in the monkey or steps15 - 19 in the rat) per testis and the number of spermatozoa per epididymis were estimated in 6 normal adult monkeys(Macaca fascicularis) and 6 normal adult SD rats on 25 um-thick methacrylate-embedded sections using a contempo-rary unbiased and efficient stereological method-the optical disector. The diameter and length of the efferent ductulesand ductus epididymidis and the volume of the epididymal fluid in the tubules were also estimated. Results: The totalnumber of the late spermatids per testis was 2902 ±749 (million, x ±s) in the monkey, or 179 ±31 in the rat; thenumber of spermatozoa per epididymis was 3235 ±1835 in the monkey, or 241±76 in the rat. Conclusion: A largenumber of spermatozoa was densely packed and stored in the ductus epididymidis; the epididymal transit time for sper-matozoa was around 5 days in monkeys or 11 days in rats.

The cytoplasmic droplet may be indicative of sperm motility and normal spermiogenesis

Although the cytoplasm of spermatids is removed at the end of spermiogenesis, a tiny portion is usually retained in the sperm flagellum, which is termed the cytoplasmic droplet （CD） in mammals. CDs are believed to play a role in sperm volume adaptation. However, we have noticed that epididymal spermatozoa that display initial （flagellation in situ） and progressive motility mostly possess CDs, whereas spermatozoa without CDs are rarely motile, suggesting that CDs have a role in motility development during sperm epididymal maturation. In the present study, we analyzed the relationship between the presence or absence of CDs, motility development and positional changes of CDs during sperm epididymal maturation in mice and monkeys. We also examined CDs on spermatozoa of three knockout mouse lines with late spermiogenic defects. Our data suggest that the CD is a normal organelle transiently present exclusively on epididymal spermatozoa, and normal CD morphology and location are associated with normal motility development during epididymal maturation of spermatozoa. Abnormal CD formation, e.g., a complete lack of CDs or ectopic CDs, is indicative of defective spermiogenesis. If CDs are essential for sperm motility development, then CDs may represent an ideal drug target for the development of non-hormonal male contraceptives.