Effect of transient scrotal hyperthermia on human sperm: an iTRAQ-based proteomic analysis.

BACKGROUND: Through this prospective study, we aimed to explore the change of molecular modification after the transient scrotal hyperthermia on human sperm. METHODS: Ten healthy subjects selected with strict screening criteria underwent testicular warming in a 43 °C water bath for 30 min a day for 10 consecutive days. Semen samples were collected 2 weeks before the first heat treatment and 6 weeks after the first heat treatment. Proteins from the samples were labeled with isobaric tags for relative and absolute quantitation and analyzed by two-dimensional liquid chromatography-tandem mass spectrometry. RESULTS: In contrast to the control, of the 3446 proteins identified, 61 proteins were deregulated: 28 were up-regulated and 33 were down-regulated. Approximately 95% of the differentially expressed proteins were found to participate in spermatogenesis, fertilization, or other aspects of reproduction. In particular, the expression of sperm motility and energy metabolism-related proteins AKAP4, SPESP1, ODF1, ODF2, GAPDHS, and ACTRT2, validated by western blotting of the proteins obtained from human and mouse samples, tended to be reduced under scrotal hyperthermia. CONCLUSIONS: The results indicated that the proteins AKAP4, ODF1, ODF2, GAPDHS, SPESP1, and ACTRT2, play an important role in the heat-induced reversible reduction in sperm concentration and motility and have the potential to be the biomarkers and clinical targets for scrotal heat treatment induced male infertility.

Antifertility effectiveness of a novel polymer matrix composite and its influence on the endometrium in rhesus macaques (Macaca mulatta).

OBJECTIVE(S): To explored the antifertility effectiveness and influence on the endometrium of a micro-copper/low-density polyethylene/methyl vinyl silicone rubber (Cu/LDPE/MVQ) composite in rhesus macaques. STUDY DESIGN: Healthy reproductive aged female rhesus macaques underwent abdominal hysterotomy for surgical placement of either the experimental Cu/LDPE/MVQ composite (Cu/LDPE/MVQ, n=5), bare copper wire (Cu, n=5), or hysterotomy only sham-operation group [(SOI, n=4), (SOII, n=6)]. Females in the Cu/LPDE/MVQ, Cu, and SOI groups were housed with fertile males for approximately three menstrual cycles. We assessed pregnancy by hysterectomy. Females in the Cu/LDPE/MVQ, Cu, and SOII groups underwent hysterectomy at about 4 months post-insertion for histologic assessment of morphologic changes of the endometrium, evaluation of materials using scanning electron microscopy (SEM), and evaluation of the inflammatory markers, including substance P receptor (SPR), associated with endometrial bleeding using enzyme linked immunosorbent assay, quantitative RT-PCR, and Western blot analyses. RESULTS: All of the SOI group females became pregnant (4/4, 100%). In contrast, no pregnancies occurred in either the Cu/LDPE/MVQ (0/5, 0%) or Cu (0/5, 0%) groups. We observed histologic features consistent with chronic endometrial inflammation in all females of the Cu group, but none of the SOII or Cu/LDPE/MVQ animals. Levels of inflammatory markers were significantly increased in the Cu group, compared with SOII or Cu/LDPE/MVQ groups (p<.05). SEM showed evidence of corrosion in the Cu wire not seen in the Cu/LDPE/MVQ group. CONCLUSION(S): Cu/LDPE/MVQ material provided a contraceptive effect similar to Cu in macaques, with a lower impact on inflammation and inflammatory markers of the endometrium. IMPLICATIONS: This study demonstrates the possibility of a Cu/LDPE/MVQ composite as an alternative to conventional copper device materials.

Efficacy of combined traditional Chinese medicine spray with premature ejaculation desensitization therapy for the treatment of primary premature ejaculation.

OBJECTIVES: We recommend a new kind of spray made from eight kinds of traditional Chinese medicine, we aimed to investigate the safety and clinical efficacy of combined traditional Chinese medicine spray (TCMS) with premature ejaculation desensitization therapy (PEDT) for the treatment of primary premature ejaculation (PPE). METHODS: A total of 90 patients with PPE were randomly assigned to receive TCMS, PEDT monotherapy or TCMS plus PEDT combination therapy for 6 weeks. Intravaginal ejaculation latency time (IELT) and Chinese index of sexual function for premature ejaculation (CIPE-5) were measured to evaluate the effect of each treatment. RESULTS: Eighty six (86) participants completed the study voluntarily. Both IELT and CIPE-5 in these three groups increased after treatment when compared with baseline levels (p< 0.01). IELT and CIPE-5 after treatment in TCMS plus PEDT group were significantly higher than those in the other two groups (both p <0.05). Additionally, clinical efficacy in TCMS plus PEDT group (89.7%) was significantly higher than in TCMS (65.5%) and PEDT group (67.9%) (p< 0.01). CONCLUSION: The self-made TCMS was safe and effective for the treatment of PPE, a combination of TCMS and PEDT therapy was more effective than the TCMS or PEDT monotherapy.

Copper nanoparticle-induced ovarian injury, follicular atresia, apoptosis, and gene expression alterations in female rats.

Numerous studies have reported the accumulation of copper nanoparticles (Cu NPs) in organs and the corresponding damage, although whether Cu NPs can be translocated to the ovaries and their ovarian toxicity are still unknown. In this study, three groups of female rats were injected with 3.12, 6.25, or 12.5 mg/kg Cu NPs for 14 consecutive days. The pathological changes, hormone levels, apoptosis and apoptotic proteins, oxidative stress, and gene expression characteristics in the ovaries were then investigated. The results demonstrated that the Cu NPs exhibited obvious accumulation in the rat ovaries, leading to ovarian injury, an imbalance of sex hormones, and ovarian cell apoptosis. Cu NP exposure activated caspase 3, caspase 8, caspase 9, and tBid, decreased the protein levels of Bcl-2, increased the expression levels of the proteins Bax and cytochrome c, and promoted malondialdehyde (MDA) accumulation and superoxide dismutase (SOD) reduction. Furthermore, gene microarray analysis showed that Cu NPs (12.5 mg/kg/d) caused 321 differentially expressed genes. Of these, 180 and 141 genes were upregulated and downregulated, respectively. Hsd17b1, Hsd3b1, Hsd3b6, and Hsd3b were involved in steroid and hormone metabolism, whereas Mt3 and Cebpb were associated with apoptosis. Overall, these findings provide strong evidence that Cu NPs trigger both intrinsic and extrinsic apoptotic pathways and regulate key ovarian genes in oxidative stress-mediated ovarian dysfunction.

Association of Alcohol Consumption with Markers of Prostate Health and Reproductive Hormone Profiles: A Multi-Center Study of 4,535 Men in China.

BACKGROUND: The effect of alcohol consumption on prostate health and reproductive hormone profiles has long been investigated and currently, no consensus has been reached. Additionally, large studies focusing on this topic are relatively rare in China. PURPOSE: To investigate the association of alcohol consumption with prostate measurements and reproductive hormone profiles in Chinese population; and to examine the relationship between hormone levels and prostate measurements. METHODS: This cross-sectional study included 4535 men from four representative provinces of China. Demographic details, family history of prostate disease, tobacco and alcohol consumption, as well as International Prostate Symptom Score (I-PSS) were collected through a questionnaire. Total prostate specific antingen (total PSA), free PSA, free PSA/total PSA ratio (f/tPSA), and reproductive hormones were measured in serum. Multi-variable regression models were used to test for association of alcohol consumption with markers of prostate health, used to test for association of alcohol consumption with reproductive hormones, and reproductive hormones with markers of prostate health. RESULTS: Alcohol consumption had no obvious impact on total PSA concentration and I-PSS. Current drinkers had lower level of free PSA (ß = -0.11, p = 0.02) and f/tPSA (ß = -0.03, p = 0.005), former drinkers also had lower level of free PSA (ß = -0.19, p = 0.02) when compared with never drinkers. Lower Luteinizing hormone (LH) (ß = -1.05, p = 0.01), sex hormone-binding globulin (SHBG) (ß = -4.71, p = 0.01) and higher estradiol (ß = 7.81, p = 0.01) was found in current drinkers than never drinkers, whereas higher LH (ß = 1.04, p = 0.04) and free testosterone (FT) (ß = 0.03, p = 0.02) was detected in former drinkers than never drinkers. Furthermore, LH was positively associated with f/tPSA (ß = 0.002, p = 0.006), SHBG was also positively related with free PSA (ß = 0.003, p = 0.003) and f/tPSA (ß = 0.0004, p = 0.01). Both total testosterone (TT) and FT were inversely related with I-PSS (OR = 0.97, 95% CI, 0.95-0.98; OR = 0.23, 95% CI, 0.11-0.45, respectively). CONCLUSIONS: Alcohol consumption could affect serum free PSA concentration and also f/tPSA ratio, and also acts as an endocrine disruptor on the male reproductive hormone profiles. LH and SHBG were positively related with fPSA and f/tPSA, and higher level of TT and FT may be helpful for improving participants' subjective symptoms.

Effect of transient scrotal hyperthermia on sperm parameters, seminal plasma biochemical markers, and oxidative stress in men.

In this experimental prospective study, we aimed to analyze the effect of transient scrotal hyperthermia on the male reproductive organs, from the perspective of sperm parameters, semen plasma biochemical markers, and oxidative stress, to evaluate whether different frequencies of heat exposure cause different degrees of damage to spermatogenesis. Two groups of volunteers (10 per group) received testicular warming in a 43°C water bath 10 times, for 30 min each time: group 1: 10 consecutive days; group 2: once every 3 days. Sperm parameters, epididymis and accessory sex gland function, semen plasma oxidative stress and serum sex hormones were tested before treatment and in the 16-week recovery period after treatment. At last, we found an obvious reversible decrease in sperm concentration (P = 0.005 for Group 1 and P= 0.008 for Group 2 when the minimums were compared with baseline levels, the same below), motility (P = 0.009 and 0.021, respectively), the hypoosmotic swelling test score (P = 0.007 and 0.008, respectively), total acrosin activity (P = 0.018 and 0.009, respectively), and an increase in the seminal plasma malondialdehyde concentration (P = 0.005 and 0.017, respectively). The decrease of sperm concentration was greater for Group 2 than for Group 1 (P = 0.031). We concluded that transient scrotal hyperthermia seriously, but reversibly, negatively affected the spermatogenesis, oxidative stress may be involved in this process. In addition, intermittent heat exposure more seriously suppresses the spermatogenesis compared to consecutive heat exposure. This may be indicative for clinical infertility etiology analysis and the design of contraceptive methods based on heat stress.

Evaluation of semen quality in 1808 university students, from Wuhan, Central China.

The aim of this study was to evaluate the semen quality of university students in Wuhan, the largest city in the world in terms of the number of university students. All student sperm donors recorded in the Hubei Province Human Sperm Bank from 1 March 2010 to 31 December 2013 were screened. At last, a total of 3616 semen samples from 1808 university student sperm donors were eligible and retrospectively analyzed. Each donor's semen parameters were averaged over two samples and compared with the World Health Organization criteria, and a generalized linear regression model was used to examine several determinants of semen quality. We found that the mean and median values were 3.0 ml and 2.8 ml for semen volume, 50.2 × 10 6 ml-1 and 50.0 × 10 6 ml-1 for sperm concentration, 148.1 × 10 6 and 142.1 × 10 6 for total sperm count, and 58.6% and 60.0% for total sperm motility. About 85.0% of donors had parameters that were all normal. Season and duration of abstinence were critical factors affecting semen quality. We also found a decrease in sperm concentration during the 4 years observation; however, this may not be a strong evidence to confirm the declining trend of semen quality. In conclusion, semen quality of university students in Wuhan was not optimal and should be paid high attention, long-term observation and further study should be carried out to confirm the present situation.

[Detecting testis- and epididymis-specific methylated promoters in human cell-free seminal DNA by MeDIP-qPCR].

OBJECTIVE: To establish a method of methyl-DNA immunoprecipitation (MeDIP)-real time quantitative PCR (qPCR) for detecting the promoter methylation level in cell-free seminal DNA (cfsDNA). METHODS: We obtained cfsDNA samples from 6 normozoospermia men (the NZ group) and 6 post-vasectomy patients (the PV group), and mixed the samples from different individuals of each group, respectively. Then we made DNA fragments by ultrasonication, separated the methylated DNA fragments by MeDIP, and determined the methylation level of the promoters in cfsDNA by qPCR. RESULTS: The methylation levels of the promoters PRAME, PEG10, MORC1, GML, HOXA5, DNMT3L, SNURF, MSH4, DAZ1 and CLPB were 14.93, 2.64, 0.69, 2.66, 17.50, 21.10, 5.98, 2.28, 13.50 and 3.86%, respectively, in the NZ group, obviously lower than 121.25, 73.62, 16.25, 42.90, 76.74, 112.40, 59.79, 25.85, 91.90 and 64.53% in the PV group. The results of MeDIP-qPCR for the methylation of PRAME, MORC1, GML, HOXA5, DNMT3L, SNURF, MSH4 and DAZ1 were coincident with the results of genome-wide promoter methylation microarray. CONCLUSION: MeDIP-qPCR can quantitatively measure the promoter methylation level in cfsDNA, and effectively determine the testis- and epididymis-specific methylated promoters in human semen.

[Advances in the studies of cell-free seminal DNA].

Cell-free DNA, also referred to as extracellular DNA, has been detected in many kinds of human body fluids, including blood plasma, urine, cerebrospinal fluid, bronchoalveolar lavage fluid, amniotic fluid, and seminal plasma. At present, cell-free DNA has been reported widely as promising noninvasive biomarkers for disease diagnosis and research. Recent years have witnessed some progress in the studies of the general characteristics of cell-free DNA, such as its concentration, extent of molecular weight, origin and existing forms, as well as in its clinical application. Cell-free seminal DNA has been proposed as promising noninvasive biomarkers for the studies and diagnosis of male idiopathic infertility, and the early diagnosis, treatment evaluation and outcome prediction of testicular germ cell tumors and prostatic cancer. This review summarizes the general characteristics and biological functions of cell-free DNA, and outlines the research status and application perspective of cell-free seminal DNA.

Comparative study on contraceptive efficacy and clinical performance of the copper/low-density polyethylene nanocomposite IUD and the copper T220C IUD.

BACKGROUND: The study was conducted to compare the antifertility effectiveness and side effects of the copper/low-density polyethylene nanocomposite IUD (experimental group) and the copper T220C IUD (control group). STUDY DESIGN: One hundred females were randomly divided into two groups (experimental group and control group, n = 50 in each group). Clinical observation and comparative study were performed on the two groups for 12 months. RESULTS: Follow-up rate was 100% at the 12th month. In the experimental group and control group, the cumulative continuation rates were both 92.0 per 100 women at the 12th month and there was no difference between them (p > .05). The pregnancy rate, removal rate and expulsion rate were low with the difference being not statistically significant (p>.05). The most common side effects were excessive menstrual bleeding, spotting and pain. The rates of side effects were lower in the experimental group than in control group, especially during the initial 3 months after insertion with the differences being statistically significant (p < .05). CONCLUSION: The new design of the copper/low-density polyethylene nanocomposite IUD showed low pregnancy rate, high contraceptive efficacy and satisfactory acceptability. The study suggested that the TCu220C IUD also had high contraceptive efficacy, but had relatively more side effects.

[Expression patterns of age-related molecule c-kit, HIWI and vimentin in the rat testis and epididymis].

OBJECTIVE: To examine the differences in the expressions of c-kit, HIWI and vimentin in the testis and epididymis of rats aged different months, investigate the regular changes in the expression of characteristic molecules in the ripening and senescent process of the testis and epididymis, and provide experimental evidence for studies on the aging of the male reproductive system. METHODS: Sprague-Dawley male rats were divided into a young group (6-month-old, n=10), an adult group (12-month-old, n=10) and an aged group (24-month-old, n=10). The immunohistochemical SP method was used to examine c-kit, HIWI and vimentin in the testis and epididymis of the rats. RESULTS: The positive immunohistochemical reaction to c-kit was observed mainly in spermatogonia, weakly in other cells and also in the epithelia and lumens of the epididymis. The abundant expression of HIWI was detected in all stages of spermatogenic cells in the testis and epididymis of the 6-month-old and 12-month-old rats. Moreover, spermatogonias and primary spermatocytes displayed intense expression. Sertoli cells, Leydig cells, myoid cells and vascular endothelial cells (VEC) were also HIWI-positive cells. But the positive expression decreased with age in the testis and epididymis of the 24-month-old rats (P > 0.05). vimentin expression was weak in the testis and epididymis of the 6-month-olds but increased significantly in the 24-month-olds (P > 0.01). CONCLUSION: The expressions of c-kit and HIWI decreased in the testis and epididymis of the 24-month-old rats, while vimentin expression increased markedly with age. The results suggest that the aging of the testis and epididymis is closely related to the abnormal transduction of cell signals.

The antifertility effectiveness of copper/low-density polyethylene nanocomposite and its influence on the endometrial environment in rats.

PURPOSE: The study was conducted to investigate the antifertility effectiveness of copper/low-density polyethylene nanocomposite (nano-Cu/LDPE) and its influence on the endometrial environment in rats. METHODS: One hundred and seventy sexually mature female Sprague-Dawley (SD) rats were randomly divided into five groups: sham-operated control group (SO group, n=10), bulk copper group (Cu group, n=40), LDPE group (n=40), and nano-Cu/LDPE groups I (n=40) and II (n=40). Twenty rats in each group except for the SO group were mated with male rats of proven fertility, from 30 days after insertion, and the antifertility rates (ATs) were observed at Day 11 of pregnancy. The concentrations of prostaglandin E(2) (PGE(2)) and tissue plasminogen activator (tPA) in the endometrium of the remaining rats in each group were measured by using ELISA at the 30th and 60th day after insertion, respectively. RESULTS: ATs in the Cu group and nano-Cu/LDPE groups I and II (100%) were significantly higher than those in the LDPE group (p<.05). Compared with those in the SO group, the concentrations of PGE(2) and tPA in all experimental groups except for PGE(2) levels in the LDPE group were significantly increased at Day 30 after insertion, and these parameters in the nano-Cu/LDPE groups were significantly lower than in the Cu group (p<.05). At Day 60 after insertion, tPA levels were still higher in the Cu and nano-Cu/LDPE groups, but there was no difference in PGE(2) levels in all groups except for the Cu group. CONCLUSION: Nano-Cu/LDPE exhibits satisfactory contraceptive efficacy with less influence on the endometrium PGE(2) and tPA levels.

[Analysis of the differences in the expression of HSP27 and c-kit between benign prostatic hyperplasia and prostatic cancer tissues].

OBJECTIVE: To examine the differences in the expression of HSP27 and c-kit between benign prostatic hyperplasia (BPH) and prostatic cancer (PCa) tissues and to analyse the relationship between their expression and BPH and PCa, especially the relationship with the occurrence, development, prognosis and treatment of PCa. METHODS: An immunohistochemical staining (SP method) for HSP27 and c-kit was undertaken on 40 BPH and 40 PCa tissues samples. RESULTS: Consistent patterns of cytoplasmic staining for HSP27 were seen in all sections of tissue from BPH. The glandular epithelium stained very strongly positively and the stroma stained positively. The staining for HSP27 in PCa tissues was located in the cytoplasm of glandular epithelia, but the expression of HSP27 in PCa was higher than BPH (P < 0.05). The staining for c-kit in BPH tissues was located in the cytoplasm of smooth muscle cells, and in PCa tissues was located in epithelial cells. The expression of c-kit in PCa tissues was lower than BPH (P < 0.05). The expression level of both HSP27 and c-kit were decreased with the development of grade of PCa (P < 0.05); HSP27 was increased with the development of clinical stage of PCa (P < 0.05 ); c-kit was decreased with the development of clinical stage of PCa (P < 0.05). CONCLUSION: The expression level of HSP27 and c-kit was highly correlated with the process of the development from BPH to PCa, and also correlated with tumor grades and stages. The expression of HSP27 and c-kit may be used as an important pathological index and may be helpful for the treatment of PCa.

[Effect of eucommia ulmoides oliv on SOD activity and alpha-actin expression in the penile tissues of diabetic rats].

OBJECTIVE: To study the effect of aqueous abstract from eucommia ulmoides oliv on the activity of superoxide dismutase (SOD) and alpha-actin expression in the penile tissues of rats with diabetes mellitus (DM) in vitro. METHODS: A diabetes model was established by administration of alloxun twice to Sprague Dawley (SD) rats. Ten diabetic and 10 normal rats were randomly selected and the penile strips of each rat were divided into four equal shares and cultured in two groups, a eucommia ulmoides oliv coculture group (Group A, further dicided into 1 microg/ml, 10 microg/ml and 100 microg/ml subgroups) and a control group (Group B). Seven days later, the activity of SOD in the culture medium was detected by spectrophotometry, and the levels of micro-actin expression in the penile tissues were examined with the immunohistochemical method. RESULTS: Compared with Group B, the activity of SOD in the culture medium in athe 10 and 100 microg/ml subgroups was notably elevated (P < 0.01), and the numbers of immunoreactive positive cells of alpha-actin in the penile tissues remarkably increased (P < 0.01). CONCLUSION: The activity of SOD and alpha-actin expression in the penile tissues of diabetic rats in vitro can be increased by eucommia ulmioides oliv.

[Effect of L-methionine on trace elements in lead-intoxicated mice].

OBJECTIVE: To investigate the effect of L-methionine (L-Met) on the content of Zn, Cu, Mn, Fe in liver, brain, spleen and kidney of lead intoxicated mice. METHODS: Distilled water was given to 10 mice (normal control group) and lead acetate solution of 400 micro g/ml Pb(2+) to 20 mice to serve as drinking water for 10 days. The lead administration was then withdrawn and lead exposed mice were randomly divided into two groups: the lead control group took distilled water as drinking water for 4 weeks to serve as positive control, the other one took L-Met solution (0.5 mg/ml) as drinking water for 4 weeks (Pb + L-Met group) to serve as the treatment group. All the animals were sacrificed on the 1st day after 4 weeks, and the contents of Zn, Cu, Fe, Mn, Pb in liver, brain, spleen and kidney were measured by Inductively Coupled Plasma (ICP) Emission Spectrometry. RESULTS: Lead contents in liver, brain, spleen and kidney of Pb control group [(1.490 +/- 1.654) micro g/g, (3.470 +/- 2.757) micro g/g, (4.975 +/- 2.993) micro g/g, (0.066 +/- 0.001) micro g/g respectively], were higher than those in normal control group [(0.015 +/- 0.001) micro g/g, (0.009 +/- 0.007) micro g/g, (0.027 +/- 0.002) micro g/g, (0.006 +/- 0.015) micro g/g, P < 0.05] while Zn contents in liver, brain, spleen and Fe and Mn content in liver, brain, spleen and kidney in Pb control group were lower than those in normal control group (P < 0.05). Pb contents of brain, spleen and Cu content of kidney in Pb + L-Met group were higher than those in normal control group (P < 0.05). Zn contents of liver, brain, spleen, Fe contents of liver, brain, spleen, kidney, and Mn contents of brain, spleen in Pb + L-Met group were lower than those in normal control group (P < 0.05). Fe contents of liver, brain, Zn content of spleen, Cu content of kidney and Mn contents of liver, brain, spleen in the Pb + L-Met group were higher than those in the Pb control group (P < 0.05). The lead levels of four organs in the Pb + L-Met group were lower than those in the Pb control group (P < 0.05). CONCLUSION: Lead could be eliminated by L-Met, which may affect the distribution and metabolism of trace elements in mice.