Increased oxidative damage of sperm and seminal plasma in men with idiopathic infertility is higher in patients with glutathione S-transferase Mu-1 null genotype

Aim： To examine whether a relationship exists between glutathione S-transferase Mu-1 （GSTM1） gene polymorphism and the susceptibility of sperm and seminal plasma from patients with idiopathic infertility to oxidative stress. Methods： Fifty-two men with idiopathic infertility and 60 healthy fertile men were recruited to this study. GSTM1 gene polymorphism was determined by polymerase chain reaction （PCR） and both the infertile and control individuals were divided into GSTM1 null and GSTM1 positive groups according to their GSTM1 gene structure. We compared reactive oxygen species （ROS） generation, malondialdehyde （MDA）, protein carbonyls and glutathione （GSH） concentrations, and glutathione S-transferase （GST） activity in seminal plasma and spermatozoa from infertile patients and controls with respect to GSTM1 genotype. Results： Significantly higher levels of oxidative stress and damage markers were found in idiopathic infertile men with the GSTM1 null genotype compared with those with the GSTM1 positive genotype. There was no significant difference in genotype distribution for theGSTM1 variant between the idiopathic infertile subjects and fertile subjects. Patients with the GSTM1 null genotype also had lower sperm concentrations than those with GSTM1 positive genotype. Conclusion： Our results suggest that the susceptibility of sperm and seminal plasma to oxidative stress is significantly greater in idiopathic infertile men with the GSTM1 null genotype compared with those possessing the gene. Therefore, in patients with idiopathic infertility, GSTM1 polymorphism might be an important source of variation in susceptibility of spermatozoa to oxidative damage.     

Male serum Chlamydia trachomatis IgA and IgG, but not heat shock protein 60 IgG, correlates with negatively affected semen characteristics and lower pregnancy rates in the infertile couple

BACKGROUND: The objective of this study was to evaluate whether serum Chlamydia trachomatis immunoglobulin-A (IgA), IgM and C. trachomatis heat shock protein 60 (CHSP60) IgG are of additional value to C. trachomatis IgG regarding the impact on fecundity in infertile couples, and to relate C. trachomatis serum antibodies to semen characteristics, diagnoses and pregnancy outcome. METHODS: A total of 226 infertile couples, previously tested for C. trachomatis IgG, were tested for C. trachomatis IgA, IgM and CHSP60 IgG, and semen samples from all men were analysed. RESULTS: Chlamydia trachomatis serum IgA in men (but not in women) correlated with reduced chances of achieving pregnancy [p = 0.021, relative risk (RR) =0.65, 95% confidence interval (CI) 0.42-1.005] and in combination with C. trachomatis IgG the chance was further reduced (p =0.001, RR = 0.35, 95% CI 0.15-0.84). Chlamydia trachomatis serum IgA was also significantly correlated with reduced motility of the spermatozoa (-8.7%, p = 0.023), increased number of dead spermatozoa (+10.5%, p = 0.014) and higher prevalence of leucocytes in semen (+122%, p = 0.005), and in combination with C. trachomatis IgG positivity, there was also a decrease in sperm concentration (-35%, p = 0.033), the number of progressive spermatozoa (-14.8%, p = 0.029) and a rise in the teratozoospermia index (+4.4%, p = 0.010). CHSP60 IgG correlated with reduced motility (-5.6%, p = 0.033), and in the women to tubal factor infertility (p = 0.033), but no correlations of C. trachomatis serum IgM or CHSP60 IgG with pregnancy rates were found. CONCLUSIONS: Chlamydia trachomatis serum IgA in the male partner of the infertile couple has an additive value to IgG in predicting pregnancy chances, and serum IgA and IgG are associated with subtle negative changes in semen characteristics.     

Current approaches for detection of carcinoma in situ testis

Testicular germ cell tumours have a favourable prognosis if detected early, but are potentially lethal in a subset of patients. Multi-modality treatment is often necessary, thus the preferable time of diagnosis is at the pre-invasive, but unfortunately often asymptomatic precursor stage of carcinoma in situ (CIS). This review describes current possible approaches for the detection of CIS. At present, an open testicular biopsy is the only definitive way of establishing the presence of CIS. The tissue section should be of an adequate size, be properly fixed, and evaluation be supported by at least one solid immunohistochemical marker, for example PLAP, OCT-3/4 or AP-2gamma. Determination of who should be offered testicular biopsies is based on clinical and ultrasonic examination along with the evaluation of risk factors. A surgical biopsy is an invasive procedure with potential complications, although rare. Therefore, a noninvasive and equally reliable method is needed. Testicular ultrasound is risk-free, painless and at present the only noninvasive method of aid for andrologists when CIS is suspected. The presence of testicular microlithiasis is, in some cases, indicative of pre-malignant changes, especially in males with additional risk factors. Promising results have recently been obtained with a novel noninvasive detection method based on immunocytological AP-2gamma-staining of CIS cells in semen. This method could be a supporting method in andrology centres where careful follow-up is possible. In conclusion, one difficulty is to determine in which males CIS should be suspected; secondly, there does not as yet exist an optimal noninvasive method of diagnosis that is more acceptable than an open surgical biopsy.     

Fertility treatment in male cancer survivors

The present study reviews the use of assisted reproductive technology in male cancer survivors and their partners. As antineoplastic treatment with chemotherapy or radiation therapy, has the potential of inducing impairment of spermatogenesis through damage of the germinal epithelium, many male cancer survivors experience difficulties in impregnating their partners after treatment. The impairment can be temporary or permanent. While many cancer survivors regain spermatogenesis months to years after treatment, some become infertile with a-, oligo- or azoospermia. An option to secure the fertility potential of young cancer patients is to cryopreserve semen before cancer treatment for later use. A desired pregnancy may be obtained in couples where the husband has a history of cancer, using assisted reproductive technology with either fresh or cryopreserved/thawed semen. Successful outcomes have been obtained with intrauterine insemination (IUI) as well as in vitro fertilization (IVF) with or without the use of intracytoplasmic sperm injection (ICSI). In conclusion, male cancer survivors and their partners who have failed to obtain a pregnancy naturally within a reasonable time frame after end of treatment should be referred to a fertility clinic.     

Semen and the diagnosis of infertility in Aristotle

Aristotle (384-322bc) was one of the leading intellectual figures of all time. In his work he systematised a massive amount of knowledge on a diverse range of subjects, including medicine. This article discusses the observations and hypotheses of this great philosopher on semen and infertility, as they are presented in his work Generation of Animals. This is combined with an evaluation of his positions in relation to those of the Hippocratic Corpus on the same subject. An extensive review of Aristotle's work Generation of Animals was performed with particular focus on his perspectives about semen and infertility. Publications referring to this work were also reviewed. According to Aristotle, semen is that which contains the principles that come from both parents when they unite. He believed that semen was formed by the secretion of nutriments by the body, developing his theories of sterility on this basic principle. A lack of fertility is attributed to genetic or acquired causes. He proposed methods for diagnosing sterility, primarily the 'water test' for men and the 'pessary' method for women. Even if his observations contain clear mistakes, such as attributing only secondary functions to male testicles and the identification of menses as women's 'seed', Aristotle's views also contain keen observations and exceptional thinking, both on the characteristics of semen and the causes of sterility (infertility).     

Tadalafil as an in vitro sperm motility stimulant

Tadalafil (Cialis) is a known oral selective phosphodiesterase-5 inhibitor used widely in the management of erectile dysfunction. To assess its ability on human sperm motility in vitro, 70 asthenozoospermic semen specimens delivered by masturbation were investigated. Semen samples were divided equally into four tubes, one as a control and to the others tadalafil dissolved solution was added in vitro in three different concentrations (4.0, 1.0, 0.5 mg ml(-1) respectively). The tubes were incubated and were followed up for sperm motility per cent changes for 0.5, 1, 2, 3 h. It was found that the concentration used played an important role in the degree of sperm enhancement. Specimens treated with 4 mg ml(-1) tadalafil solution demonstrated a significant decrease in sperm motility compared with the controls. Specimens treated with 1.0 mg ml(-1) solution demonstrated significant increase in sperm progressive forward motility. Specimens treated with 0.5 mg ml(-1) solution demonstrated significant increases in sperm motility but lower than that of 1 mg ml(-1) concentration. It is concluded that in vitro use of tadalafil solution in special concentration has a significant stimulatory effect on asthenozoospermic sperm motility.     

A new method to estimate quantitatively seminal vesicle and prostate gland contributions to ejaculate

AIMS: We sought to optimize a quantitative noninvasive method to determine the concentration in their glands of origin of biochemical markers of compartments of the male genital tract as the first step towards validation of a novel method for estimation of drug concentrations in these male genital tract compartments. METHODS: Sixty-eight men participated. We compared four collection devices to split ejaculate into fractions. Fractions were assayed for fructose and prostate specific antigen (PSA) as unique markers of the seminal vesicle and prostate, respectively. Seminal vesicle fructose and prostatic PSA were estimated using a linear regression method, based on fructose-PSA axis intercepts, and compared with an older method which solves a simultaneous series of equations. RESULTS: A five-compartment collection device performed best with mean (95% confidence interval) PSA vs. fructose r(2) of 0.84 (0.71, 0.98, P < 0.001). Using resampling simulations, glandular PSA and fructose estimates were highly variable and often implausible when using only two fractions. Using our method, the prostate contributed 37-44% to the whole ejaculate and the seminal vesicle contributed 55-61%. The novel regression method was highly correlated (r(2) > or = 0.98) with older methods. CONCLUSIONS: We developed a noninvasive quantitative method of male genital tract biochemical marker estimation using a five-compartment tray to collect three to five ejaculate fractions. Our novel regression method is quantitative and more fully developed than older methods. This noninvasive method for determining glandular marker concentrations should be useful to provide quantitative estimates of drug concentrations in these glands.     

微粉化对马钱子溶出度的影响

目的 考察微粉化时制马钱子中士的宁、马钱子碱溶出度的影响.方法 采用《中华人民共和国药典》2010年版二部附录溶出度浆法,用HPLC测定,分别在人工胃液和人工肠液(无酶)模拟体内环境的溶出介质下,转速50r/min,体积为900 mL,比较微粉化前后马钱子粉的溶出行为.结果 以T50、Td、T90为溶出度参数,在人工肠液中,以马钱子碱、士的宁为考察指标,马钱子超微粉分别为0.38、1.33、19.50,0.39、1.31、17.63 min;马钱子粗粉分别为19.43、44.53、249.12,20.84、44.96、229.95 min.在人工胃液中,以马钱子碱、士的宁为考察指标,马钱子超微粉分别为0.32、1.25、20.60,0.45、1.41、20.81 min;马钱子粗粉分别为20.53、43.31、220.30,20.76、46.02、241.82 min,两者比较差异均有统计学意义(P＜0.01).结论 微粉化能提高马钱子中士的宁、马钱子碱的溶出量与溶出速度.     

短尾小鼠脊柱发育形态与病理生理学研究

目的:研究脊柱发育异常疾病模型─短尾小鼠生长发育中的病理生理学特征,并与正常B6小鼠比较,为动物模型开发与应用提供有价值资料。方法:短尾小鼠和B6小鼠分别行脊髓的病理切片和电镜切片观察,且测定2种小鼠的心电图数据和精液品质数据进行比较分析。结果:短尾小鼠脊髓外观形态完整,但神经元密度低于正常B6小鼠;短尾小鼠脊髓超微结构有异常现象;2种小鼠平均心率比较差异有统计学意义(P<0.05〉,短尾小鼠P-R间期比B6小鼠略长,QRS群间期、Q-T间期较短,两者比较差异无统计学意义(P>0.05);短尾小鼠精子活力偏低,两者比较差异无统计学意义(P>0.05)。结论:短尾小鼠脊髓灰质的前角运动神经元数目略有减少;短尾小鼠髓鞘和线粒体结构存在异常;短尾小鼠心电图有异常现象;短尾小鼠生殖力偏低与精液品质无明显关系。     

人精液中NO与TP、Alb及Tf含量的关系

目的 探讨人精液一氧化氮（NO）与总蛋白（TP）、白蛋白（Alb）和转铁蛋白（Tf）含量的关系。方法参照WHO标准方法，进行精液常规分析。采用镀铜镉还原荧光法检测N0代谢产物硝酸盐（NO3^-），用双缩脲法检测TP，溴甲酚绿法测定Alb，用免疫散射比浊法检测Tf含量。结果 不育活率异常组、少精子和无精子症组的NO含量明显高于正常生育组，TP、Alb和Tf含量明显低于正常生育组，两组之间存在高度显著性差异（P〈0.01），不育组NO含量与TP、Alb和Tf含量呈显著性负相关（r=-0．88、r=-0．98、r=-0．68，P均〈0．05）。结论 精液蛋白质含量测定有助于精子质量的评价，对男性不育症的诊治有一定的指导意义，NO对精子运动能力及蛋白质的分泌利用有抑制作用，这对不育症的机制研究有重要价值。