Decreased testosterone secretion index and free testosterone level with multiple symptoms for late-onset hypogonadism identification: a nationwide multicenter study with 5980 aging males in China.

Late-onset hypogonadism (LOH) is a syndrome in middle-aged and elderly men caused by age-related testosterone deficiency. Age-related change of total testosterone (TT) of Asian males is different from Caucasian population, suggesting difference for LOH identification in Asians. A nationwide cross-sectional study involving six centers in China was conducted. Totally 6296 men aged 40-79 were recruited. After exclusions 5980 men were left for analyses. The serum TT level, was neither decreased with aging nor correlated with most hypogonadal symptoms. Instead, ten hypogonadal symptoms were found to be significantly correlated with free testosterone and testosterone secretion index, thus were chosen to form a concise scale. Further analysis identified a level of free testosterone <210 pmol/L, testosterone secretion index <1.8, and the concise scale score ≧17 could be diagnosed as having significantly aggravated LOH. This study developed an evidence-based criteria for LOH identification in Chinese population and may be adopted in other Asians. It includes the impaired testosterone secretion ability and deficiency of bioavailable testosterone, which should be the main cause in LOH pathogenesis despite normal TT levels, as well as correlated multiple hypogonadal symptoms. Our results may guide the LOH treatment to increase testicular function of testosterone secretion and bioavailable testosterone.

Decline in semen concentration of healthy Chinese adults: evidence from 9357 participants from 2010 to 2015.

The present study aims to analyze sperm concentration trends among young and healthy Chinese adults in Wuhan, Central China, from 2010 to 2015. Semen analysis data from 9357 participants were collected and analyzed using a general linear model and the Cochran-Armitage trend test. A significant decline was observed in sperm concentration (ß [standard deviation]: -1.53 [0.16]; P < 0.001). In addition, a decline in sperm density was observed by stratifying student versus nonstudent sperm donors and by analyzing the year of birth or birth year cohort of the participants. Furthermore, the percentage of participants with sperm densities of over 40 × 106 ml-1 significantly decreased with year. Notably, a dramatic decline in sperm density was recorded over the first 5 years of study. This research reported a decline in sperm concentration among young adults in Wuhan, Central China, in 2010-2015.

Prevalence and risk factors of lower urinary tract symptoms in Chinese adult men: a multicentre cross-sectional study.

There has been no previous population-based study reporting the prevalence and risk factors of male lower urinary tract symptoms (LUTS) among men in mainland China. This cross-sectional study was conducted from 2013 to 2014 in three representative provinces of China: Guangdong, Hubei and Jiangsu. 3250 individuals participated in the interviews, which involved a questionnaire covering sociodemographic characteristics, lifestyle, dietary patterns and the International Prostate Symptom Score (IPSS). Blood was collected for lipids, glucose, insulin and reproductive hormone tests. The incidences of LUTS and its obstructive and irritative symptoms were calculated. Risk factors for LUTS were identified using multivariable logistic regression analysis. The prevalence of moderate to severe LUTS and its obstructive and irritative symptoms was 14.3%, 13.1% and 16.1%, respectively, and increased with age. The prevalence in Guangdong was much lower than that in Hubei and Jiangsu in different ages. Increased fasting plasma glucose and decreased HDL-C levels were associated with an increased risk of moderate to severe LUTS (OR = 1.30, 95% CI: 1.02-1.65 and OR = 2.06, 95% CI: 1.08-3.94, respectively). Free testosterone < 0.22 ng/ml decreased the risk of moderate to severe LUTS and obstructive and irritative symptoms by about 20-30%. An inadequate daily intake of vegetables, fruit and water significantly increased the risk of LUTS by 1.3-to 2.0 times. In conclusion, the prevalence of LUTS in Chinese men is high and increases with age. Dietary patterns may be critical for the development of LUTS. Thus, dietary modifications could be a useful strategy for preventing the development of LUTS.

The plasma miR-125a, miR-361 and miR-133a are promising novel biomarkers for Late-Onset Hypogonadism.

Circulating miRNAs have been shown to serve as diagnostic/prognostic biomarkers in cancers and other diseases. However, the role of plasma miRNAs in Late-onset hypogonadism (LOH) diagnosis is still unknown. Using Illumina HiSeq2000 sequencing at discovery phase, and then two-step validated by reverse transcriptase polymerase chain reaction (RT-PCR) assays in verification phases. We verified that the expression levels of miR-125a-5p, miR-361-5p and miR-133a-3p were significantly altered in LOH group compared to the control group. The area under the receiver operating characteristic (ROC) curve (AUC) is 0.682, 0.698 and 0.765, respectively. The combination of three miRNAs showed a larger AUC (0.835) that was more efficient for the diagnosis of LOH. Among three miRNAs, miR-133a-3p had the best diagnostic value for LOH with 68.2% sensitivity and 77.3% specificity. Regression analyses show that miR-133a-3p level was negatively associated with the ageing males' symptoms (AMS) scale. However, miR-361-5p level was positively associated with serum testosterone concentrations. In summary, plasma miRNAs are differentially expressed between LOH and healthy controls. We validated three miRNAs that could act as novel biomarkers for diagnosis of LOH. These miRNAs may be involved in the development of LOH. However, further large and functional studies are warranted to confirm our findings.

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.

Analysis of the Screening Results for 3,564 Student Sperm Donors in Hubei Province, China.

OBJECTIVE: To determine the reasons for the elimination of student sperm donors after semen analysis, physical examination, and laboratory tests in the Hubei Provincial Human Sperm Bank. Understanding the status of student sperm donors can provide a valid reference for the screening work of sperm banks. STUDY DESIGN: The screening data from 3,564 student sperm donors in Hubei Provincial Human Sperm Bank from January 1, 2010-April 30, 2013, were retrospectively analyzed. RESULTS: A total of 733 students (20.57%) qualified for semen analysis in the human sperm bank, whereas 602 students (16.89%) completed the sperm donation procedure. The main reasons for elimination were as follows: unqualified semen parameters (2,748 cases), failed semen extraction (83 cases), sexually transmitted diseases (44 cases), hereditary or chromosomal disorders (44 cases), and hepatitis B infection (25 cases). Education level and temperature/climate possibly affect semen quality. CONCLUSION: Unqualified semen parameters were the main reason for elimination among student sperm donors. Human sperm banks should promote reproductive health knowledge and information on improving semen quality among students when promoting sperm donation.

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.

n-Butanol extract of Rhynchosia volubilis Lour: a potent spermicidal agent In Vitro.

Rhynchosia volubilis Lour has been a major drug in a folk prescription for contraception in China, whereas its mechanism remains unknown. Its antifertility effects on male mice and antimicrobial activities on sexually transmitted infection (STI) pathogens were previously reported. This study was undertaken to develop the n-Butanol extract of Rhynchosia volubilis Lour (BERVL) as a spermicidal agent with STI prevention. The spermicidal activities of BERVL with different doses were assessed using selected high-motile sperms of normal human semen samples, and their inhibitory effects on Lactobacillus acidophilus were determined. The mechanism of the spermicidal activity was explored by aqueous Eosin Y and Hoechst 33342/PI staining. The results showed spermicidal activities and inhibitory effects of BERVL on Lactobacillus acidophilus were dose-dependent. Dose of 90 mg/mL BERVL terminated all progressive sperm motility within 2 min, and had slight inhibitory effect on Lactobacillus acidophilus, suggesting it was an effective and safe dose for contraception use. About 80% sperms exposed to BERVL displayed changes consistent with high permeability of head membrane. It is concluded that BERVL as spermicide has advantages over N-9 with strong ability to instantaneously kill human sperm and possesses light inhibitory effect on Lactobacillus acidophilus.

Variability in the morphologic assessment of human sperm: use of the strict criteria recommended by the World Health Organization in 2010.

OBJECTIVE: To determine the variability in the recognition of normal sperm and various sperm defects using the strict criteria recommended by the World Health Organization (5th edition, 2010). DESIGN: Sperm morphologic assessment by three experienced evaluators. SETTING: Image processing laboratory and reproduction research institute. PATIENT(S): Semen donors from a sperm bank. INTERVENTION(S): The morphology of 5,296 sperm was evaluated using statistical analyses of variability. MAIN OUTCOME MEASURE(S): The proportion and coefficients of variation (CVs) of normal sperm, defects of specific parts, and the categories of defects were measured. The degree of agreement between any two of the three evaluators was calculated. The multiple anomalies index, teratozoospermia index, sperm deformity index, and the CVs were also measured. RESULT(S): The CVs of normal sperm, multiple anomalies index, teratozoospermia index, and sperm deformity index were 4.80%, 4.14%, 5.75%, and 6.81%, respectively. A broader range (4.80%-132.97%) of CVs was observed for the recognition of various defects. The coefficients of the degree of agreement concerning specific morphologic parts of sperm varied (0.387-0.607), with lower relative values for the head and mid-piece than for the tail and cytoplasm. CONCLUSION(S): The sperm head is more difficult to evaluate than the other parts using the criteria recommended by the World Health Organization in 2010. The degree of agreement concerning specific parts and various defects varied in broad ranges. A stricter definition for each defect is needed.

[Screening results and causes of uncompleted donation process in 1 145 sperm donors].

OBJECTIVE: To improve the reception and recruitment of sperm donors in sperm banks in China, and solve the problem of insufficiency in sperm donation. METHODS: We reviewed the recruitment of 1 145 men for sperm donation in the Human Sperm Bank of Hubei Province from September 2011 to April 2012, analyzed the reasons for those not included, and interviewed those included but unwilling to donate sperm. RESULTS: Among the 1 145 recruits, 551 (48.12%) were students and 594 (51.88%) were other individuals. After the first semen screening, 503 (43.93%) quitted, including 202 students (36.66% of the students recruited) and 301 others (50.67% of the other individuals recruited). After the second semen screening, 432 (37.73%) were excluded, and another 45 (3.93%) excluded after laboratory examination, including 16 cases of mycoplasma positive. Totally, 165 recruits (14.41%) passed the semen screening and laboratory examination, but only 144 of them (87.27%) completed, while the other 21 (12.73%) failed to complete the whole donation process. CONCLUSION: Low rates of screening qualification and donation process completion are common problems in human sperm banks. The rate of qualified sperm donors can be increased and the operational cost of the human sperm bank can be reduced by enabling the recruits to accomplish the whole donation process. Explanation at the reception, later interview with the recruits, and donors' trust in the sperm bank play important roles in raising the completion rate of sperm donation process.

[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.

Characterization of a piRNA binding protein Miwi in mouse oocytes.

Argonaute proteins and Piwi proteins bind with microRNA (mRNA) and Piwi-interacting RNA (piRNA), respectively, to form functional complexes. Piwi proteins are mostly restricted to germ cells and stem cells, and the Piwi-piRNA pathway is required for normal spermatogenesis. Although piRNAs were also recently identified in mammalian oocytes, expression of Piwi proteins in the ovary has not been well characterized. Previous studies did not detect mRNA of Miwi, a murine homologue of Piwi proteins, in total RNA of mouse ovary tissue. We demonstrated herein the presence of Miwi in murine oocytes. Reverse transcription polymerase chain reaction (RT-PCR), Western blot, and immunofluorescence based on quantum dots immune labeling technique were used to investigate the expression profile of Miwi in oocytes of adult and neonatal females at 0, 1, 2, 3, and 4 weeks postpartum. Although RT-PCR was negative in total RNA of the adult ovary, both RT-PCR and Western blot detected Miwi in oocytes of adult mice, and ovaries of neonatal females. Miwi transcript and protein peaked at 1 and 2 weeks postpartum, respectively. Miwi mRNA was detectable in newborn mouse ovaries, implying its transcription was initiated at least in the primordial follicle. Its protein was strong in late primary and secondary follicles, but appeared to decrease as maturation proceeded. The exclusion of anti-Miwi immunofluorescence from some cytoplasmic granules was observed. Given that diverse biologic and molecular functions have been revealed for the Piwi-piRNA pathway in germline cells of many species, Miwi might be an important functional protein in murine folliculogenesis.

Cell-free seminal mRNA and microRNA exist in different forms.

BACKGROUND: The great interest in cell-free mRNA, microRNA (miRNA) as molecular biomarkers for clinical applications, and as 'signaling' molecules for intercellular communication highlights the need to reveal their physical nature. Here this issue was explored in human cell-free seminal mRNA (cfs-mRNA) and miRNA (cfs-miRNA). METHODOLOGY/PRINCIPAL FINDINGS: Selected male reproductive organ-specific mRNAs, miRNAs, and piRNAs were quantified by quantitative real-time PCR in all experiments. While the stability of cfs-miRNA assessed by time-course analysis (up to 24 h at room temperature) was similar with cfs-mRNA, the reductive changes between cfs-miRNA and cfs-mRNA after filtration and Triton X-100 treatment on seminal plasma were very different, implying their different physical nature. Seminal microvesicles (SMVs) were then recovered and proportions of cfs-mRNA and cfs-miRNA within SMVs were quantified. The amounts of SMVs- sequestered cfs-mRNAs almost were the same as total cfs-mRNA, and were highly variable depending on the different sizes of SMVs. But most of cfs-miRNA was independent of SMVs and existed in the supernatant. The possible form of cfs-miRNA in the supernatant was further explored by filtration and protease K digestion. It passed through the 0.10-µm pore, but was degraded dramatically after intense protease K digestion. CONCLUSIONS/SIGNIFICANCE: The predominant cfs-mRNA is contained in SMVs, while most cfs-miRNA is bound with protein complexes. Our data explained the stability of extracellular RNAs in human semen, and shed light on their origins and potential functions in male reproduction, and strategy of developing them as biomarkers of male reproductive system.

Immunization of male mice with B-cell epitopes in transmembrane domains of CatSper1 inhibits fertility.

OBJECTIVE: To evaluate the contraceptive ability of two B-cell epitopes in CatSper1. DESIGN: In vivo study with mice. SETTING: University laboratory animal service center. ANIMAL(S): BALB/c mice. INTERVENTION(S): Two predicted B-cell epitopes in the extracellular part of transmembrane domains and pore region of CatSper1 were synthesized to immunize male mice. MAIN OUTCOME MEASURE(S): Fertility, epididymal sperm function, and the presence of specific antibody in immunized males were investigated. RESULT(S): Significant reduction of fertility was observed in mating trial with no evident systemic illness or abnormal mating behavior. Epididymal sperm of epitope-immunized males exhibited impaired ability to fertilize eggs in vitro, and showed sperm agglutination in some animals, while presenting no changes in sperm viability or progressive motility. High titer of antibodies was induced in the sera, and the antibodies' specificity was confirmed. The binding of the antibodies to epididymal sperm of epitope-immunized males was observed. CONCLUSION(S): CatSper members could be the effective and viable targets for immunocontraception. These two epitopes in CatSper1 share high identity between mouse and human and may be effective for fertility regulation in humans.

[Survey on knowledge, attitude, practice related to contraception among college students in Beijing].

OBJECTIVE: To understand the current status and relative factors on knowledge, attitude, practice to contraception among college students in Beijing and to provide evidence for developing aimed contraceptive service and education in colleges. METHODS: Quantitative surveys were employed, questionnaire was conducted anonymously among 12,450 unmarried college students under informed consent. RESULTS: Among 12,129 students, acknowledgement of contraceptive knowledge among females was superior to males in college (P=0.000). Rates on the favor for unmarried sexual practice and 'sexual liberation' were 81.9% and 60.2% for males, comparing to 74.8% and 54.3% for females respectively. The prevalence rates for masturbation were 74.3% for males and 24.9% for females respectively. 18.4% of the male and 10.5% of the female students had ever experienced unmarried sexual intercourse. Among those students who had sexual experiences, 29.4% of the males had made their sex partners pregnant and 23.1% of the female students had experienced unexpected pregnancy. Statistical significance was found between males and females (P=0.000) in terms of attitudes and behaviors. 51.3% of the male and female students took contraceptive measures during the episode of first sexual intercourse. The excuses for not taking any measures would include: sexual intercourse happened incidentally, not interested in using condom, only one sexual experience would not cause pregnancy, etc. CONCLUSION: The prevalence of unmarried sexual intercourse among college students had been rising. Among those who had experienced unmarried sexual intercourse, the rate of taking contraceptive measures was low and the incidence of unexpected pregnancy was high. It is urgent to strengthen the education and service programs on contraception among college students.

Inhibition of human sperm function and mouse fertilization in vitro by an antibody against cation channel of sperm 1: the contraceptive potential of its transmembrane domains and pore region.

OBJECTIVE: To explore the contraceptive potential of the CatSper1 transmembrane domains and pore region in vitro. DESIGN: In vitro study with human sperm and mouse fertilization. SETTING: Andrology laboratory of an academic research center. PATIENT(S) AND ANIMAL(S): Normozoospermia and viripotent BALB/c mice. INTERVENTION(S): The specific binding of an anti-CatSper1 IgG antibody (H-300) to CatSper1 was confirmed by Western blot and immunofluorescence. Sperm from humans and mice were incubated with H-300. MAIN OUTCOME MEASURE(S): The effects of H-300 on human sperm progressive motility, abnormal acrosome, hyperactivated motility, and mouse in vitro fertilization rates were analyzed. RESULT(S): A significant decline in sperm progressive motility was observed after 1, 2, and 4 hours of incubation with H-300; the change was mainly ascribed to the decline of fast progressive motility. Significant inhibition of the hyperactivated motility was observed after 5 hours of incubation with H-300. The incubation of mouse sperm with H-300 before insemination reduced the in vitro fertilization rate to 28% of control levels (72% inhibition). CONCLUSION(S): CatSper1 may be a potential target for immunocontraception, and the antibody may be a tool to study the function of ion channels in sperm in which relatively fewer methods can be applied.

Inhibition of the urokinase-type plasminogen activator by triplex-forming oligonucleotides in rat Sertoli cells: a new contraceptive alternative?

Urokinase-type plasminogen activator (uPA), expressed in Sertoli cells in the testis, is closely related with tight junctions of blood-testis barrier (BTB), and it has been considered as a potential contraceptive target. In the present study, the antigene effects of triplex-forming oligodeoxynucleotides (TFO) targeting uPA in rat Sertoli cells were investigated in vitro. The stable triplexes, formed by uPA specific TFOs under physiological conditions, were tested by means of electrophoretic mobility shift assays (EMSA). Although tPA, another form of plasminogen activators (PAs), partially compensated the lose of PAs activities, uPA mRNA and protein were significantly reduced as demonstrated by real-time reverse transcription PCR and a chromogenic assay, after the treatment of Sertoli cells with uPA specific TFOs at a concentration of 330 nM. The capacity of TFOs resistance to nuclease degradation was enhanced by the phosphorothioated on the backbone of the oligonucleotides. Our results indicated that the TFOs can downregulate uPA expression and uPA might be an alternative contraceptive target.

[Effect of urokinase-type plasminogen activator on the mitochondrial membrane potential of mouse capacitated-spermatozoa in vitro].

OBJECTIVE: To study the mechanism of uPA improving sperm capacitation by investigating the effect of uPA on the mitochondrial function of mouse capacitated-sperm in vitro. METHODS: Mitochondrial function of mouse capacitated-spermatozoa was evaluated through the assessment of mitochondrial membrane potential using JC-1 performed by flow cytometer and fluorescent microscope respectively. The experiment and the control groups were designed according to the presence or absence of uPA, each divided into 5 subgroups based on the different time of uPA treatment (or BWW in the control groups) at 0, 5, 15, 30 and 60 min respectively. RESULTS: (1) Compared with that at 0 min, the mean fluorescence intensity of JC-1 within the spermatozoal body and the percentage of orange-red colored spermatozoa in the experiment group were increased significantly at 5 and 15 min respectively after uPA incubation (P < 0.05). (2) The mean fluorescence intensity of JC-1 within the spermatozoal body at 15, 30 and 60 min and the percentage of orange-red colored spermatozoa at 5 and 15 min in the group were significantly higher than those in the control group (P < 0.05). CONCLUSION: uPA could increase the mitochondrial membrane potential of mouse capacitated-spermatozoa in vitro, and maintain it at a high level for a certain period of time. By enhancing sperm mitochondrial function, uPA may provide sufficient energy for capacitated-spermatozoa to increase their motility and change their motor pattern, which might be one of the therapeutic mechanisms of uPA on male infertility.

[Urokinase-type plasminogen activator and urokinase-type plasminogen activator receptor in the seminal plasma and sperm of fertile and oligoasthenozoospermia males].

OBJECTIVE: To find the difference between the levels of urokinase-type plasminogen activator (uPA) and urokinase-type plasminogen activator receptor(uPAR) in the seminal plasma and sperm of fertile and oligoasthenozoospermia men, and to understand their correlation with male fertility. METHODS: The levels of uPA in the seminal plasma and sperm of 22 normospermic males and 44 oligoasthenozoospermia patients were measured by ELISA. RESULTS: (1) The average level of uPA in the seminal plasma and sperm of the normospermic group, ([4803.69 +/- 602.78] mU/L) and ([30.29 +/- 3.16] mU/10(6) sperm) were higher than those of the oligoasthenozoospermia group, ([4061.35 +/- 736.23] mU/L), and ([20.51 +/- 4.2] mU/10(6) sperm) (P < 0.01). (2) The average level of uPAR in the sperm of the normospermic group ([12.97 +/- 3.11] mU/10(6) sperm) was significantly higher than that of the oligoasthenozoospermia group, ([6.09 +/- 1.45] mU/10(6) sperm) (P < 0.01). (3) The levels of uPA and uPAR in the sperm and the content of uPA in the seminal plasma were positively correlated with sperm motility and viability. CONCLUSION: Urokinase-type plasminogen activator is related with male fertility and the levels of uPA and uPAR vary in the seminal plasma and sperm of fertile and oligoasthenozoospermia males.

[Detection of Trichomonas vaginalis with direct immunofluorescence assay].

OBJECTIVE: To detect Trichomonas vaginalis (Tv) by direct immunofluorescence assay (DFA) and discuss its clinical significance. METHODS: The parasites at a concentration of 3 x 10(5) cells/L were fixed by acetone on slides which were then blocked by 1% BSA (bovine serum albumin) or 10% BSA or 10% NCS (newborn calf serum) respectively, incubated with different dilution of polyclonal goat anti-Tv IgG (1:20-1:2,560) for different incubation time (15, 30, 45, 60, 90, 120 min). 120 clinical vaginal specimens were examined by direct immunofluorescence assay, the wet mount method and the in vitro cultivation. RESULTS: Blocked by 1% or 10% BSA, incubated at 37 degrees C for 45 min with a titer 1:160 of polyclonal antibody were the optimal conditions for direct immunofluorescence assay. Its sensitivity and specificity were 87.9% and 98.6% respectively in comparison with the in vitro cultivation method. CONCLUSION: Direct immunofluorescence assay is a useful alternative to the wet mount method which shows a lower sensitivity.