Expression of Attractin in Male Reproductive Tract of Human and Mice and Its Correlation with Male Reproduction

The expression of Attractin mRNA and protein in testis and semen of human and male mice was investigated. Human testis and semen samples were all collected from Reproductive Center of Reumin Hospital, Wuhan University in December, 2012. Testis samples were collected from 7 cases of obstructive azoospermias when they were subjected to diagnosed testis biopsy, and 30 nor- mal human semen samples were obtained from those cases of semen analysis. Adult mice testis tis- sues were obtained from 10 2-month-old male BALB/c mice, and 60 male mice at different ages were classified into 10 groups （day 1, 5, 10, 15, 21, 28, 35, 42, 56, and 120 respectively, n=6 each）. The expression of Attractin mRNA and protein in testis was detected by RT-PCR and Western blotting re- spectively. Human semen samples were centrifuged into sperm plasma （SP） and sperm extract （SE）, and mice sperm samples were collected from the epididymis of 10 adult male BALB/c mice. Western blotting was used to determine the Attractin protein expression level. Attractin mRNA and protein were expressed in the testis of both patients with obstructive azoospermias and adult Bcl/B mice. Quantitative RT-PCR revealed that no Attractin mRNA was detectable in day 1 male BALB/c mice group. The Attractin mRNA and protein levels were low on the day 10, and increased with age until day 56. On the day 120, the expression levels of Attractin were decreased. As for human semen sam- pies, Attractin protein was expressed in both SP and SE, but didn＇t exist in samples from the epidi- dymis of male BALB/c mice. It was suggested that Attractin acted as a novel active substance and was involved in male reproduction in both human and BALB/c mice, but it exerted a different ex- pression profile in different mammal species.

Persistent organic pollutants and male reproductive health

Environmental contaminants such as persistent organic pollutants （POPs） are man-made bioaccumulative compounds with long half-lives that are found throughout the world as a result of heavy use in a variety of consumer products during the twentieth century. Wildlife and animal studies have long suggested adverse effects of exposure to these compounds on human reproductive health, which, according to the endocrine disrupter hypothesis, are ascribed to the compounds＇ potential to interfere with endocrine signaling, especially when exposure occurs during certain phases of fetal and childhood development. An extensive number of epidemiological studies have addressed the possible effects of exposure to POPs on male reproductive health, but the results are conflicting. Thus far, most studies have focused on investigating exposure and the different reproductive health outcomes during adulthood. Some studies have addressed the potential harmful effects of fetal exposure with respect to malformations at birth and/ or reproductive development, whereas only a few studies have been able to evaluate whether intrauterine exposure to POPs has long-term consequences for male reproductive health with measurable effects on semen quality markers and reproductive hormone levels in adulthood. Humans are not exposed to a single compound at a time, but rather, to a variety of different substances with potential divergent hormonal effects. Hence, how to best analyze epidemiological data on combined exposures remains a significant challenge. This review on POPs will focus on current knowledge regarding the potential effects of exposure to POPs during fetal and childhood life and during adulthood on male reproductive health, including a critical revision of the endocrine disruption hypothesis, a comment on pubertal development as part of reproductive development and a comment on how to account for combined exposures in epidemiological research.

3ioinformatics for spermatogenesis： annotation of male ＇eproduction based on proteomics

Proteomics strategies have been widely used in the field of male reproduction, both in basic and clinical research. Bioinformatics methods are indispensable in proteomics-based studies and are used for data presentation, database construction and functional annotation. In the present review, we focus on the functional annotation of gene lists obtained through qualitative or quantitative methods, summarizing the common and male reproduction specialized proteomics databases. We introduce several integrated tools used to find the hidden biological significance from the data obtained. We further describe in detail the information on male reproduction derived from Gene Ontology analyses, pathway analyses and biomedical analyses. We provide an overview of bioinformatics annotations in spermatogenesis, from gene function to biological function and from biological function to clinical application. On the basis of recently published proteomics studies and associated data, we show that bioinformatics methods help us to discover drug targets for sperm motility and to scan for cancer-testis genes. In addition, we summarize the online resources relevant to male reproduction research for the exploration of the regulation of spermatogenesis.

A scoping review of SARS-CoV-2 and male infertility: Concerns and future prospects

Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has been detected in the blood,urine,facial/anal swabs,semen,and vaginal discharge;all have been shown to contain SARSCoV-2 RNA.Recent findings have highlighted the prospect of SARS-CoV-2 invading the genital system in addition to other tissues,which might give rise to reproductive concerns.This investigation sheds light on male reproductive tract vulnerability to invasion by SARS-CoV-2 and provides a foundation for further researches into male fertility.Males are infected with COVID-19 at a higher rate than females.As a result,some data suggest that this viral infection might affect the male reproductive system.The probable causes for male genital tract abnormalities in COVID19 are:(1)high expression of angiotensin-converting enzyme 2 in the testes;(2)SARS-CoV-2 infection indirectly induces immune response in the testes;(3)SARS-CoV-2 directly damages male genital cells by virus-receptor binding activity;(4)fever in SARSCoV-2 infected males may cause damages to testicular cells;(5)testosterone level decreased in SAR-CoV-2 infected males;(6)males are more susceptible to COVID-19 than females,which may be due to differences in the physiology of the genital tract.This review seeks to offer some insights into the potential causes of COVID-19 that affect the male reproductive system,as well as future prospect on this issue.

Male Reproductive Toxicology

The objective of this paper is to discuss recent developments in the application of biological markers to animal models of male reproductive toxicology. We have divided this paper into three major sections: First, a discussion of the testing protocols currently under investigation by the National Toxicology Program (NTP) and the EPA's Health Effects Research Laboratory; second, an examination of what we consider to be important and practical biological markers available to investigators to assess the effects of toxicants on the male reproductive system; and third, a discussion of promising new technologies, such as molecular and immunological probes, and in vitro techniques using isolated and cultured cells, which in the future may be exploited for the development of additional biological markers of male reproductive toxicity. Where appropriate we have made specific recommendations for the use of these biological markers in animal protocols and have pointed out those noninvasive markers which have application to screening human males.

Reproductive Biology of the Male Sheath-Tailed Bat,Taphozous longimanus(Emballonuridae) From India

The seasonal chronological events of the repreductive cycle in the male sheath-tailed bat T. longimanus and changes in structure and function of accessory sex organs were studied at Varanasi (latitude: 25° N, 83°E). The first peak of spermatogenesis was ob served in September with the appearance of spermatozoa. The spermatogenic arrest in win ter dormancy (December) was followed by a second peak of spermatogenesis in January.This perbo coincides with ovulation in females. The third peak of spermatogenesis started after regression in late April, and completely regressed tubules were found after May. The cycle of accessory sex gland complex generally paralleled the testicular cycle, reaching maxi mum hypertrophy from September to January and again in late April. Thereafter, the ac cesory sex glands involuated. Spermatozoa were present in the cauda epididymidis through out the year. The reproductive cycle in family Emballonuridae varies with changes in lati tude, and this was so even in members of the same species. The presence of spermatozoa in the cauda epididymidis throughout the year is an interesting feature in the reproductive cy cle. Temperature and photoperiod do not play a regulatory role in reproduction in T.longimanus

Aggressive angiomyxoma of the epididymis: A case report

BACKGROUND Aggressive angiomyolipoma is an extremely rare benign mesenchymal tumor that was originally described as a locally recurrent mucinous spindle cell tumour.Agg-ressive angiomyolipoma originates from myofibroblasts,vascular smooth muscle cells,or fibroblasts,and displays various phenotypes of myofibroblasts and abnor-mal muscle arteries.Aggressive angiomyolipoma was first identified in 1983 and fewer than 50 male patients have been reported to date.It is an extremely rare mesenchymal tumour and often confused with other diseases.Patients with epididymal aggressive angiomyolipoma lack typical symptoms,most of which occur incidentally,although some patients may experience mild pain,discomfort,and swelling.Pain may be exacerbated by pressure from the mass.CASE SUMMARY A 66-year-old male was admitted to the hospital on January 14,2022 with chief complaint of swelling in the left scrotum for one year.There was no apparent cause for the swelling.The patient did not consult with any doctor or receive any treatment for the swelling.The enlarged scrotum increased in size gradually until it reached approximately the size of a goose egg,and was accompanied by discom-fort and swelling of the left cavity of the scrotum.The patient had no history of any testicular trauma,infection,or urinary tract infection.The patient urinated freely,1-2 times at night,without urgency,dysuria(painful urination),or haematuria.There was no significant family history of malignancy.The patient underwent excision of the enlarged tumour and the left epididymis under general anaes-thesia on January 18,2022.Twelve months of follow-up revealed no recurrence.The patient was satisfied with the treatment.CONCLUSION Aggressive angiomyolipoma is extremely rare clinically and often confused with other diseases.The pathogenesis of aggressive angiomyolipoma is unclear and the clinical presentation is mostly a painless enlarged mass.The diagnosis of aggressive angiomyolipoma requires a combination of medical history,preoperative imaging such as computed tomography and magnetic resonance imaging,cytological examination and preoperative and postoperative pathological biopsy.The preferred treatment is surgery,with the possibility of a new alternative treatment option after hormonal therapy.Aggressive angiomyolipoma should be considered in the differential diagnosis of parametrial tumors of the male genital area that present as clinically significant masses.The high recurrence rate of aggressive angiomyolipoma may be related to incomplete tumor resection,and patients with aggressive angiomyolipoma are advised to undergo annual postoperative follow-up and imaging for recurrence.

Long-term exposure to aryl hydrocarbon receptor agonist neburon induces reproductive toxicity in male zebrafish(Danio rerio)

Neburon is a phenylurea herbicide that is widely used worldwide,but its toxicity is poorly studied.In our previous study,we found that neburon has strong aryl hydrocarbon receptor(AhR)agonist activity,but whether it causes reproductive toxicity is not clear.In the present study,zebrafish were conducted as a model organism to evaluate whether environmental concentrations of neburon(0.1,1 and 10μg/L)induce reproductive disorder in males.After exposure to neburon for 150 days from embryo to adult,that the average spawning egg number in high concentration group was 106.40,which was significantly lower than 193.00in control group.This result was mainly due to the abnormal male reproductive behavior caused by abnormal transcription of genes associated with reproductive behavior in the brain,such as secretogranin-2a.The proportions of spermatozoa in the medium and high concentration groups were 82.40%and 83.84%,respectively,which were significantly lower than 89.45%in control group.This result was mainly caused by hormonal disturbances and an increased proportion of apoptotic cells.The hormonal disruption was due to the significant changes in the transcription levels of key genes in the hypothalamus-pituitary-gonadal axis following neburon treatment.Neburon treatment also significantly activated the AhR signaling pathway,causing oxidative stress damage and eventually leading to a significant increase in apoptosis in the exposed group.Together,these data filled the currently more vacant profile of neburon toxicity and might provide information to assess the ecotoxicity of neburon on male reproduction at environmentally relevant concentrations.

Bazi Bushen alleviates reproductive aging in aged male mice

Bazi Bushen(BZBS),a traditional Chinese medicine(TCM),has demonstrated therapeutic efficacy in testicular dysfunction within D-galactose and NaNO_(2)mouse models.This study aimed to ascertain if BZBS could also mitigate the decline in testicular function associated with natural aging.Therefore,male aged mice were employed to evaluate the preventive effects of BZBS on male reproductive aging.This was achieved by assessing sex hormone production,testicular histomorphology,and spermatogenesis.Relative to the untreated aged control group,BZBS administration elevated the levels of sex hormones and spermatocyte populations and preserved normal testicular structure in aged mice.Notably,spermatogenesis was maintained.Further analyses,including malondialdehyde(MDA)assays and real-time PCR,indicated that BZBS diminished testicular oxidative stress and the inflammatory burden.Corroborating these findings,mice treated with BZBS exhibited reductions in the populations of senescent and apoptotic cells within the seminiferous tubules,suggesting alleviated cellular damage.In contrast,we observed that rapamycin,a drug known for its longevity benefits,induced excessive testicular apoptosis and did not decrease lipid peroxidation.Collectively,our results highlight BZBS’s promising clinical potential in counteracting male reproductive aging,underlining its mechanisms of action.

The effect of environmental contaminants on testicular function

Male reproductive health has deteriorated considerably in the last few decades. Nutritional, socioeconomic, lifestyle and environmental factors （among others） have been attributed to compromising male reproductive health. In recent years, a large volume of evidence has accumulated that suggests that the trend of decreasing male fertility （in terms of sperm count, quality and other changes in male reproductive health） might be due to exposure to environmental toxicants. These environmental contaminants can mimic natural oestrogens and target testicular spermatogenesis, steroidogenesis, and the function of both Sertoli and Leydig cells. Most environmental toxicants have been shown to induce reactive oxygen species, thereby causing a state of oxidative stress in various compartments of the testes. However, the molecular mechanism（s） of action of the environmental toxicants on the testis have yet to be elucidated. This review discusses the effects of some of the more commonly used environmental contaminants on testicular function through the induction of oxidative stress and apoptosis.

An Association of Elevated Serum Prolactin with Phthalate Exposure in Adult Men

Objective To investigate the associations of hormone circulation with phthalate exposure in adult men. Methods Semen and serum samples were collected from 118 men who were suspected of infertility. Phthalate diesters including dibutyl phthalate （DBP） and diethylhexyl phthalate （DEHP） in both semen and serum samples were measured, along with serum levels of follicle stimulating hormone （FSH）, luteinizing hormone （LH）, testosterone （T）, estradiol （E 2 ） and prolactin （PRL）. Results Serum PRL was positively associated with serum DBP and DEHP and semen DEHP in all models of Spearman correlation, linear regression and binary logistic regression. In linear regression models adjusted for potential confounders and excluding subjects with undetectable phthalates, a 10‐fold increase in semen DEHP was associated with a 23% increase in serum PRL, as well as a 26% increase in serum DBP and a 20% increase in serum DEHP. In logistic regression models all subjects demonstrated a dose‐response relationship between above reference value PRL and semen DEHP （odds ratio per tertile adjusted for potential confounders = 1.0, 1.70, 3.50; P for trend = 0.01）, and serum DBP （1.0, 1.10, 2.62; P for trend = 0.04）, and serum DEHP （1.0, 1.46, 4.69; P for trend 0.01）. A positive correlation between serum estradiol and semen DEHP （linear regression）, and an inverse correlation between semen DBP and serum testosterone and T：E 2 ratio （Spearman correlation） were also established. Conclusions Serum PRL is suggested to be positively associated with both DBP and DEHP exposure in adult men.

Does exposure to computers affect the routine parameters of semen quality?

Aim： To assess whether exposure to computers harms the semen quality of healthy young men. Methods： A total of 178 subjects were recruited from two maternity and children healthcare centers in Shanghai, 91 with a history of exposure to computers （i.e., exposure for 20 h or more per week in the last 2 years） and 87 persons to act as control （no or little exposure to computers）. Data on the history of exposure to computers and other characteristics were obtained by means of a structured questionnaire interview. Semen samples were collected by masturbation in the place where the semen samples were analyzed. Results： No differences in the distribution of the semen parameters （semen volume, sperm density, percentage of progressive sperm, sperm viability and percentage of normal form sperm） were found between the exposed group and the control group. Exposure to computers was not found to be a risk factor for inferior semen quality after adjusting for potential confounders, including abstinence days, testicle size, occupation, history of exposure to toxic substances. Conclusion： The present study did not find that healthy： men exposed to computers had inferior semen quality. （Asian J Andro12005 Sep; 7： 263-266）

Effects of plants and plant products on the testis

For centuries, plants and plant-based products have been used as a valuable and safe natural source of medicines for treating various ailments. The therapeutic potential of most of these plants could be ascribed to their anticancer, antidiabetic, hepatoprotective, cardioprotective, antispasmodic, analgesic and various other pharmacological properties. However, several commonly used plants have been reported to adversely affect male reproductive functions in wildlife and humans. The effects observed with most of the plant and plant-based products have been attributed to the antispermatogenic and/or antisteroidogenic properties of one or more active ingredients. This review discusses the detrimental effects of some of the commonly used plants on various target cells in the testis. A deeper insight into the molecular mechanisms of action of these natural compounds could pave the way for developing therapeutic strategies against their toxicity.

Exploring the reproductive ecology of the tropical semifossorial snake Ninia atrata

Based on histological analyses and field studies,this research describes the reproductive ecology of a population of Ninia atrata snakes inhabiting an oil palm plantation.Furthermore,through a multivariate approach,we explored the main drivers of reproductive output in N.atrata.Results showed that prey abundance and food intake were crucial variables contributing to reproductive output.Multiple linear regression models showed that neonates had high sensitivity(R2=55.29%)to extreme changes in climate,which was strongly related to slug and snail abundance variability and microhabitat quality.Reproductive cycles were markedly different between the sexes,being continuous in males and cyclical in females.Despite this variation,reproductive cycles at the population level were seasonal semi-synchronous.Constant recruitment of neonates all year,multiple clutches,high mating frequency,and continuous sperm production characterized the reproductive phenology of N.atrata.In addition,a significant number of previtellogenic females presented oviductal sperm as well as uterine scars,suggesting a high precocity in the species.The main drivers of reproductive output also differed between the sexes.In females,clutch size and secondary follicle variability were highly related to stomach bolus volume,fat body area,and body mass.In males,height of piles of palm leaves and body mass,rather than intrinsic reproductive traits,were the main drivers of sperm production.Nevertheless,in both cases,the relationship between body mass,prey abundance,and food intake suggests that N.atrata follows the income breeding strategy to compensate for reproductive costs and to maximize fitness.

Cigarette Smoking is Associated with Decreased Sperm Density

Objectives To study the association between cigarette smoking and sperm density in men of reproductive age. Methods We enrolled 224 male employees of a modern petrochemical plant in Nanjing, China. These men had no prior history of infertility or other reproductive diseases. Epidemiologic data, including information on smoking and other occupational and lifestyle exposures were obtained by a questionnaire interview. Semen specimens were collected from each participant and analyzed according to the WHO guidelines. Regression analyses were performed to estimate the effect of smoking on sperm density. Results Approximately 67% of the subjects had ever smoked cigarettes. Different measurements of smoking behavior were each associated with decreased sperm density. There was a significant dose-response trend between the tertites of total smoking amount in pack-years and sperm density. As compared to men who never smoked, current smokers had a significant reduction in sperm density (-13.3×106/ml; 95% CI, - 24. 1, -2. 5) ,while ex-smokers had only a small decrement in sperm density ( -2. 6×106/ml; 95%CI, -18. 7,13. 5). Starting smoking at less than 20 years of age was associated with significant reduction in sperm density (- 14.8×106/ml; 95% CI, - 27. 4, -2. 2). Starting smoking at 20 years or older was associated with a slightly smaller decrease ( -10.1×106/ml; 95% CI,-21.7,1.4). Conclusions Cigarette smoking is associated with decreased sperm density, showing an evident dose-response trend in this population.

Human ribonuclease 9,a member of ribonuclease A superfamily,specifically expressed in epididymis,is a novel sperm-binding protein

To explore the functions of human ribonuclease 9(RNase 9),we constructed a mammalian fusion expression vector pcDNA-hRNase9,prepared recombinant human RNase 9-His fusion protein from HEK293T cells and determined its N-terminal amino acid sequences.According to the determined mature protein,recombinant human RNase 9 was prepared in E.coli.Ribonucleolytic activity and antibacterial activity of recombinant human RNase 9 were detected,and the distribution of human RNase 9 on tissues and ejaculated spermatozoa and in vitro capacitated spermatozoa were analyzed via indirect immunofluorescence assay.The results showed that recombinant human RNase 9 did not exhibit detectable ribonucleolytic activity against yeast tRNA,but exhibited antibacterial activity,in a concentration/time dependent manner,against E.coli.Immunofluorescent analyses showed that the predicted human RNase 9 was present throughout the epididymis,but not present in other tissues examined,and human RNase 9 was also present on the entire head and neck regions of human ejaculated spermatozoa and in vitro capacitated spermatozoa.These results suggest that human RNase 9 may play roles in host defense of male reproductive tract.

Effects of soybean isoflavones on reproductive parameters in Chinese mini-pig boars

Background： Soybean isoflavones are structurally similar to mammalian estrogens and therefore may act as estrogen agonists or antagonists. However, it has not been determined if they have any negative effects on reproductive parameters in male livestock. Therefore, the objective of this study was to evaluate the effects of soybean isofiavones on male reproduction using Chinese mini-pig boars as a model. Fifty Xiang boars were randomly divided into five groups and fed diets containing 0, 125, 250, or 500 ppm soybean isoflavones or 0.5 ppm diethylstilbestrol for 60 days. Results： Dietary supplementation with 250 ppm of soy isoflavones markedly increased the testis index （P 〈 0.05）, fructose content （P 〈 0.05）, and o-glycosidase content in testicular tissue （P 〈 0.01）, as well as increased the number of viable germ cells （P 〈 0.01） and the level of Bcl-2 protein （P 〈 0.01）. However, 500 ppm of soybean isoflavones significantly reduced both testis and epididymis indexes （P 〈 0.0S） and lactate dehydrogenase levels （P 〈 0.01）, as well as reduced serum LH and testosterone levels （P 〈 0.05）. High levels of soybean isoflavones also increased malondialdehyde levels （P 〈 0.05）, as well as increased the numbers of early and late apoptotic germ cells （P 〈 0.01） and the level of Bax proteins （P 〈 0.05） in the testis. Conclusions： The results of this study indicate that consumption of soy isoflavones at dietary levels up to 250 ppm did not adversely affect reproductive parameters in Chinese mini-pig boars whereas higher levels of soy isoflavones may adversely affect male reproduction.

Innate and adaptive immune responses in male and female reproductive tracts in homeostasis and following HIV infection

The male and female reproductive tracts are complex microenvironments that have diverse functional demands. The immune system in the reproductive tract has the demanding task of providing a protective environment for a fetal allograft while simultaneously conferring protection against potential pathogens. As such, it has evolved a unique set of adaptations, primarily under the influence of sex hormones, which make it distinct from other mucosal sites. Here, we discuss the various components of the immune system that are present in both the male and female reproductive tracts, including innate soluble factors and cells and humoral and cell-mediated adaptive immunity under homeostatic conditions. We review the evidence showing unique phenotypic and functional characteristics of immune cells and responses in the male and female reproductive tracts that exhibit compartmentalization from systemic immunity and discuss how these features are influenced by sex hormones. We also examine the interactions among the reproductive tract, sex hormones and immune responses following HIV-1 infection. An improved understanding of the unique characteristics of the male and female reproductive tracts will provide insights into improving clinical treatments of the immunological causes of infertility and the design of prophylactic interventions for the prevention of sexually transmitted infections.

Juvenile hormone regulates photoperiod-mediated male reproductive diapause via the methoprene-tolerant gene in the ladybeetle Harmonia axyridis

Juvenile hormone (JH) absence induces photoperiod-mediated reproductive diapause, which is characterized by reproductive cessation. Although the role of methoprene-tolerant (Met)-mediated JH signaling in photoperiod-mediated female reproduction has been well documented, its role in male reproduction remains unclear. In this study, we investigated the role of JH in regulating photoperiod-mediated development of the male internal reproductive system (IRS) in the predatory ladybeetle Harmonia axyridis (Pallas). In a previous study, we found that adult male H. axyridis reared under either a short-day (SD) or long-day (LD) photoperiod had obvious differences in IRS development, but we were unable to identify the regulators of male reproductive diapause. In this study, we found that beetles reared under an SD photoperiod had significantly lower JH titer and a relatively undeveloped male IRS compared with those reared under an LD photoperiod. Additionally, application of the JH analog (JHA) methoprene promoted IRS development. Furthermore, Met knockdown strongly blocked JH signaling in males reared under the LD photoperiod, thereby slowing IRS development. Moreover, exogenous JHA did not reverse the suppressed development of the male IRS caused by Met knockdown. These results indicate that photoperiod regulates male IRS development in H. axyridis through a conserved Met-dependent JH signaling pathway.