RESUMEN
BACKGROUND: This study observes changes in levels of seminal nitric oxide (NO), nitric oxide synthase (NOS), macrophage migration inhibitory factor (MIF), sperm DNA integrity, chromatin condensation and Caspase-3in adult healthy men after scrotal heat stress (SHS). METHODS: Exposure of the scrotum of 25 healthy male volunteers locally at 40-43°C SHS belt warming 40 min each day for successive 2 d per week. The course of SHS was continuously 3 months. Routine semen analysis, hypo-osmotic swelling (HOS) test, Aniline blue (AB) staining, HOS/AB and terminal deoxynucleotidyl transferase-mediated d UDP nick-end labeling (TUNEL) were carried out before, during and after SHS. Seminal NO and NOS contents were determined by nitrate reduction method. The activated Caspase-3 levels of spermatozoa and MIF in seminal plasma were measured by the enzyme-linked immunosorbent assay (ELISA) method. Statistical significance between mean values was determined using statistical ANOVA tests. RESULTS: The mean parameters of sperm concentration, motile and progressive motile sperm and normal morphological sperm were significantly decreased in groups during SHS 1, 2 and 3 months compared with those in groups of pre-SHS (P<0.001). Statistically significant differences of sperm DNA fragmentation, normal sperm membrane, and Caspase-3 activity as well as the level of NO, NOS and MIF in semen were observed between the groups before SHS and after SHS 3 months and the groups during SHS 1, 2 and 3 months (P<0.001). After three months of the SHS, various parameters recovered to the level before SHS. WBC in semen showed a positively significant correlation with the levels of NO, NOS, MIF and Caspase-3 activity. The percentage of abnormal sperm by using the test of HOS showed a positively significant correlation with that of HOS/AB. CONCLUSIONS: The continuously constant SHS can impact the semen quality and sperm DNA and chromatin, which may be contributed to the high level of NO, NOS, MIF and Caspase-3 by SHS.
Asunto(s)
Caspasa 3/metabolismo , Trastornos de Estrés por Calor/genética , Trastornos de Estrés por Calor/metabolismo , Factores Inhibidores de la Migración de Macrófagos/metabolismo , Óxido Nítrico Sintasa/metabolismo , Semen/metabolismo , Espermatozoides/metabolismo , Adulto , Fragmentación del ADN , Voluntarios Sanos , Calor , Humanos , Masculino , Óxido Nítrico , Análisis de Semen , Recuento de Espermatozoides , Motilidad EspermáticaRESUMEN
PURPOSE: To observe changes in semen parameters, sperm DNA integrity, chromatin condensation and cysteinyl aspartate-spicific proteinases (Caspase-3) in adult healthy men after scrotal heat stress (SHS). METHODS: The scrotums of 19 healthy male volunteers were exposed to the condition of 40-43 °C SHS belt warming 40 min each day for successive 2 days per week. The course of SHS was continuously 3 months. Routine semen analysis, hypo-osmotic swelling (HOS) test, eosin Y (EY) staining sperm HOS and chromatin dispersion (HOS/SCD) test, HOS and aniline blue (HOS/AB) staining test were carried out before, during and after SHS. The activated Caspase 3 levels of spermatozoa were determined with a microtiter plate reader. RESULTS: The mean parameters of sperm concentration, motility and normal morphological sperm were significantly decreased in groups with sperm being collected during SHS 1, 2 and 3 months when compared with those in groups of pre-SHS (P < 0.01). Statistically significant differences of sperm DNA fragmentation, normal sperm membrane and vitality, and Caspase-3 activity were observed between the groups of before SHS and after SHS 3 months and the groups of during SHS 1, 2 and 3 months (P < 0.001). Three months the SHS stopped, various parameters recovered to the level before SHS. Abnormal sperm with HOS/AB and HOS/SCD showed a negatively significant correlation with normal sperm by HOS/EY test, and WBC in semen showed a positively significant correlation with Caspase-3 activity. The percentage of abnormal sperm by using the test of HOS/SCD showed a positively significant correlation with that of HOS/AB. CONCLUSIONS: The continuously constant SHS can impact the semen quality, sperm DNA integrity, chromatin condensation and Caspase-3, and the combination of HOS plus AB test may simultaneously determine the integrity of membrane and chromatin condensation at the same spermatozoon.
Asunto(s)
Caspasa 3/metabolismo , Cromatina/patología , Daño del ADN , Trastornos de Estrés por Calor/fisiopatología , Infertilidad Masculina/etiología , Escroto/patología , Espermatozoides/patología , Adulto , Estudios de Casos y Controles , Cromatina/genética , Trastornos de Estrés por Calor/complicaciones , Humanos , Infertilidad Masculina/enzimología , Infertilidad Masculina/patología , Masculino , Pronóstico , Escroto/enzimología , Análisis de Semen , Espermatozoides/enzimologíaRESUMEN
BACKGROUND: The aim of this study was to explore a new three-dimensional, reticular intrauterine device (3-DRIUD) composed of nitinol and silicone rubber and to observe the contraceptive effect of the device in rats. STUDY DESIGN: Two contraceptive experiments were performed. In the first, female rats underwent bilateral placement of a 20.0-35.0-mm 3-DRUID (experimental group, n=30) via an abdominal incision or a sham operation with no IUD (control group, n=30). Two weeks after the operation was performed, the rats from either group were caged together with male rats. The contraceptive effects of the 3-DRIUD were observed at 1 to 3 months postoperation, after which the 3-DRIUDs were removed. One month after this second operation, the rats from the two groups were again coupled with fertile male rats. In a second experiment, female rats underwent bilateral placement of a 10.0-mm 3-DRUID (n=5) via an abdominal incision or a two-dimensional IUD (2-DIUD, n=20) and mated 1 month after surgery. The single-pipeline IUD was placed in 10 rats, while the enfolded-pipeline IUD was placed in 10 different rats. RESULTS: In the first experiment, none of the females in the experimental 3-DRIUD group became pregnant (0/30, 0%) after 3 months, compared to 28/30 (93.3%, p<.0001) rats in the control group. After the 3-DRIUDs were removed from the experimental group after 3 months, 27/30 (90%) became pregnant, compared with 29/30 (97%, p>.05). The litter size (mean±SD) did not differ between groups (10.9±1.5 3-DRUID, 11.2±1.1 control, p>.05). In the second experiment, five rats had a 10.0-mm 3-DRUID (which was one third the length of one uterine horn) inserted into the bilateral uterine horns, and three of the five rats became pregnant. All 20 rats were pregnant 1 month after the insertion of the 2-DIUD. Thus, the contraceptive rate for the 2-DIUD group was 0. CONCLUSIONS: The primary contraceptive mechanism effect of the new 3-DRIUD in rodents appears to be a result of occupying physical space in the uterus.
Asunto(s)
Dispositivos Intrauterinos , Aleaciones , Animales , Fenómenos Químicos , Femenino , Infertilidad Femenina/prevención & control , Expulsión de Dispositivo Intrauterino , Dispositivos Intrauterinos/efectos adversos , Tamaño de la Camada , Ensayo de Materiales , Níquel , Embarazo , Ratas , Ratas Wistar , Elastómeros de Silicona , Titanio , Perforación Uterina/prevención & controlRESUMEN
OBJECTIVE: To investigate the breakpoints of the azoospermia factor c (AZFc) microdeletion in Chinese Han population. METHODS: We detected 9 sequence tagged sites (sY84, sY86, sY127, sY134, sY152, sY145, sY255, sY254 and sY157) to confirm AZFc microdeletions in the Y chromosome for patients with severe oligozoospermia or non-obstructive azoospermia by multiplex polymerase reaction. To locate the breakpoints of AZFc microdeletions, we analyzed 192 patients with sY255, sY254 and sY157 dele- ted by detecting sY1191, sY1197, sY1054, sY1125 and sY1206, respectively. RESULTS: Five breaking patterns were found in the 192 patients with sY255, sY254 and sY157 deleted, among which the common ones were sY1197(+), sY1191(-), sY1054(-), sY1206(-) and sY1125(+), which accounted fro 54.17% (104/192), sY1197(+), sY1191(+), sY1206(-), sY1054(-) and sY1125(+), which constituted 28.12% (54/192), sY1197(+), sY1191(-), sY1206(-), sY1054(+) and sY1125 (+), which made up 14.58% (28/192). The proximal breakpoint located between sY1197 and sY1191 was 70.83% of AZFc microdeletions, and the distant breakpoint located between sY1054 and sY1125 was 82.29%. CONCLUSION: There are 5 breaking patterns of AZFc microdeletions in Chinese Han population, the proximal and distant breakpoints mostly located at the replicons b2 and b4, respectively.
Asunto(s)
Azoospermia/genética , Cromosomas Humanos Y , Infertilidad Masculina/genética , Pueblo Asiatico/genética , Deleción Cromosómica , Humanos , Masculino , Lugares Marcados de SecuenciaRESUMEN
The aim of this study was to explore sperm chromosomal aneuploidy and DNA integrity in infertile patients with spinal cord injury (SCI). Semen samples were collected from 12 infertile men with SCI by percutaneous vasal sperm aspiration (PVSA) and from 14 male SCI patients by penile vibratory stimulation (PVS). These semen samples as well as samples from 16 donors were analyzed using the hypo-osmotic swelling (HOS) test, the sperm chromatin dispersion test, terminal deoxynucleotidyl transferase-mediated terminal uridine nick-end labeling assay, and multicolor fluorescence in situ hybridization with probes specific for the chromosomes 13, 18, 21, X, and Y. There were significant differences in the percentages of motile sperm, normal morphologic sperm, normal HOS/eosin staining, and sperm DNA fragmentation between the infertile men with SCI and the control group (P < .05 and P < .01). The sperm forward motility was significantly greater in the PVSA group than in the PVS group (P < .01). The number of round cells per milliliter of semen obtained from the 14 SCI patients by PVS was between 1 million and 12 million. The rate of sperm DNA fragmentation, as identified by the sperm chromatin dispersion test, was higher in the PVS group than in the PVSA group (P < .05). The aneuploidy rates for the SCI patients were 1.5- to 1.6-fold higher for chromosomes 13, 18, and 21, and were 2.3- to 2.4-fold higher for chromosomes X and Y than for patients in the control group (P < .001). These results suggest that for men with SCI, the semen quality is poorer, the prevalence of abnormal HOS/eosin staining is greater, and sperm DNA fragmentation and sperm chromosomal aneuploidies are seen at a higher rate compared with healthy, fertile, and normospermic men.
Asunto(s)
Infertilidad Masculina/etiología , Pene/inervación , Análisis de Semen , Recuperación de la Esperma , Espermatozoides/patología , Traumatismos de la Médula Espinal/complicaciones , Adulto , Aneuploidia , Estudios de Casos y Controles , Forma de la Célula , Distribución de Chi-Cuadrado , Cromosomas Humanos Par 13 , Cromosomas Humanos Par 18 , Cromosomas Humanos Par 21 , Cromosomas Humanos X , Cromosomas Humanos Y , Fragmentación del ADN , Humanos , Hibridación Fluorescente in Situ , Etiquetado Corte-Fin in Situ , Infertilidad Masculina/genética , Infertilidad Masculina/patología , Infertilidad Masculina/fisiopatología , Masculino , Motilidad Espermática , Traumatismos de la Médula Espinal/genética , Traumatismos de la Médula Espinal/patología , Traumatismos de la Médula Espinal/fisiopatología , Succión , VibraciónRESUMEN
PURPOSE: To explore sperm chromosomal aneuploidy, sperm membrane and DNA integrity in infertile patients with anejaculation. METHODS: Semen samples were collected from 18 infertile men with spinal cord injury (SCI) by penile vibratory stimulation (PVS) and from 14 psychogenic anejaculation (PA) patients by percutaneous vasal sperm aspiration (PVSA). These semen samples as well as samples from 16 donors were analyzed using the hypo-osmotic swelling (HOS) test, the sperm chromatin dispersion (SCD) test and multi-color fluorescence in situ hybridization (FISH) with probes specific for chromosomes 13, 18, 21, X and Y. RESULTS: There were significant differences in the percentages of motile sperm, normal morphologic sperm and sperm DNA fragmentation between the infertile men with SCI and the control group (P < 0.05 and P < 0.01). The sperm motility was significantly greater in the PA-PVSA group than in the SCI-PVS group (P < 0.01). The number of round cells per mL of semen obtained from the 18 SCI patients by PVS was between 1 and 8 million. The rate of sperm DNA fragmentation in the SCI-PVS group was higher than that of the PA-PVSA group (P < 0.05). The aneuploidy rates for the SCI patients were 2.4-fold higher for chromosomes 13, 18 and 21 and 2.2-fold higher for chromosomes X and Y than for patients in the control group (P < 0.0001). CONCLUSIONS: The semen quality is poorer, sperm DNA fragmentation and sperm chromosomal aneuploidies are seen at a higher rate for SCI patients compared to healthy, fertile and normospermic men. Whether the difference in yield is due to increased scrotal temperature, genitourinary infection, or other reasons requires further study.
Asunto(s)
Aneuploidia , Infertilidad Masculina/genética , Motilidad Espermática , Espermatozoides/citología , Cromosomas Humanos/genética , Fragmentación del ADN , Humanos , Hibridación Fluorescente in Situ , Infertilidad Masculina/etiología , Masculino , Traumatismos de la Médula Espinal/complicacionesRESUMEN
The aim of the present study is to analyze sperm membrane changes and the spermicidal effect in treatment with the crude extract from Polygala tenuifolia Willd (PTW) in vitro. The root of PTW was extracted in distilled water. Normal human spermatozoa were used to assess the spermicidal activity (Sander-Cramer assay) of the extract from the PTW root. The hypo-osmotic swelling (HOS) test and the eosin Y (EY) staining were used to detect the integrity of sperm membrane and vitality. The sperm chromatin dispersion (SCD) test was performed to determine sperm DNA integrity. N-9 was used as a reference standard and semen added to physiological saline was used as the control. Semen samples were donated by 42 healthy fertile men. The crude extract from the root of PTW could immobilize and kill 100% spermatozoa within 20 s in vitro at the concentrations of 20.0 and 10.0 mg/ml; at the concentration of 5.0 mg/ml, spermatozoa were immobilized in (39.5±3.2) s. In the groups of the crude extract from the root of PTW and N-9 solution, the rate of the normal HOS (tails swollen) and the white head (unstained) was 0%, and the rate of the abnormal HOS (tails unswollen) and red head (stained) was 100%. Sperm DNA fragmentation showed no change in exposure to the crude extract from the root of PTW and N-9 solution. The sperm revival test did not show any spermatozoa that recovered their motilities. The rapid spermicidal activity of the crude extract from the root of PTW in vitro may occur by the disruption of the sperm membrane integrity.
Asunto(s)
Extractos Vegetales/farmacología , Polygala , Espermatozoides/efectos de los fármacos , Mezclas Complejas/farmacología , Fragmentación del ADN/efectos de los fármacos , Humanos , Concentración de Iones de Hidrógeno , MasculinoRESUMEN
Mean Shift algorithm is a robust approach toward feature space analysis and it has been used wildly for natural scene image and medical image segmentation. However, high computational complexity of the algorithm has constrained its application in remote sensing images with massive information. A fast image segmentation algorithm is presented by extending traditional mean shift method to wavelet domain. In order to evaluate the effectiveness of the proposed algorithm, multispectral remote sensing image and synthetic image are utilized. The results show that the proposed algorithm can improve the speed 5-7 times compared to the traditional MS method in the premise of segmentation quality assurance.
Asunto(s)
Algoritmos , Interpretación de Imagen Asistida por Computador/métodos , Telemetría/métodos , Reconocimiento de Normas Patrones Automatizadas/métodosRESUMEN
BACKGROUND: The aim of this study was to observe the contraceptive effectiveness and reversibility of the oviduct plug with nickel-titanium (Ni-Ti) shape memory alloy and silicone rubber in rabbits. STUDY DESIGN: The oviduct plugs with Ni-Ti shape memory alloy and silicone rubber were made as contraceptive devices. The frame of the oviduct plug is made of silicone rubber and Ni-Ti shape memory alloy wire. The central part of the frame resembles a circular cylinder and the amphi-terminal of the frame is like a spigot which is formed with three mounting rings. Thirty-five New Zealand adult female rabbits were randomly divided into three groups. Thirty rabbits in Group 1 were used to test the contraceptive effect of the oviduct plug. Ten rabbits in Group 2 were used to assess the reproductive reversibility of the rabbits; 3 months after the operation, the oviduct plugs were removed from the junction of the uterus and tubes by incision of the abdomen. Another five rabbits (Group 3) served as the control group. In 30 female rabbits (Group 1), the plugs were inserted into the oviducts via a 0.5-cm incision of the uterus by using an inserter that contained the plug, then the female rabbits copulated with male rabbits once a week after the operation for 1 month. The plugs were then taken out from 10 rabbits (Group 2) 3 months after the operation. RESULTS: Out of 30 cases (Group 1), only one rabbit from Group 1 was pregnant 2 months after the insertion of the plugs due to improper insertion. However, five rabbits in the control group were all pregnant. All 10 rabbits from Group 2 were pregnant 1 month after the plug was removed. CONCLUSION: The oviduct plug with Ni-Ti shape memory alloy and silicone rubber is a new, effective and reversible contraceptive tool.
Asunto(s)
Anticoncepción/métodos , Dispositivos Anticonceptivos Femeninos , Níquel , Oviductos/cirugía , Elastómeros de Silicona , Titanio , Animales , Femenino , Masculino , Embarazo , Conejos , Distribución AleatoriaRESUMEN
OBJECTIVE: To explore the mechanism of spermicidal effect of crude extract and platycodin-D from Platycodon grandiflorum (PG) root in vitro. METHODS: Between February 2006 and December 2009, 38 fertile and healthy adult males were selected as donors. PG root was extracted and platycodin-D purified. Grouping was as follows: crude extract from PG root, platycodin-D, nonoxynol-9 (N-9, as a reference standard) and semen-added physiological saline (as control). Spermicidal experiments were carried out in vitro (Sander-Cramer test). The hypo-osmotic swelling (HOS) test and modified Eosin-Giemsa (EG) staining were used to detect the integrity of sperm membrane. Four types of sperm morphology were divided through HOS-EG test: Type A: spermatozoa with swelling in tails and head white staining HOS(+)-EG(-) (membrane intact); Type B: spermatozoa with no swelling in tails (membrane-damaged) and head white staining HOS(-)-EG(-); Type C: spermatozoa with tail swelling and head red HOS(+)-EG(+); Type D: spermatozoa with no swelling in tails and head red HOS(-)-EG(+). Sperm chromatin dispersion (SCD) test was performed to determine the integrity of sperm DNA. RESULTS: The crude extract from PG root could immobilize and kill 100% spermatozoa within 20 s in vitro at the concentrations of 50.0 g/L and 20.0 g/L (v:v = 1:1 in semen). When the semen sample was exposed to the concentrations of 2.0 g/L and 1.0 g/L of platycodin-D, all spermatozoa were immobilized within 20 s. In the control group, the mean percentage of Types A, B, C and D was (69.0 ± 8.3)%, (3.4 ± 0.5)%, (10.2 ± 1.7)% and (17.4 ± 2.1)% respectively. In the groups of platycodin-D and N-9 solution, the rate of Types A and B was 0. The rate of Types C [(65.3 ± 3.8)%] and D [(34.7 ± 7.1)%] significantly increased versus control in the platycodin-D group (P < 0.01). Sperm DNA fragmentation had no change upon an exposure to the extract from PG root, platycodin-D and N-9 solution. And the sperm revival test showed none of the spermatozoa recovered their motility. CONCLUSION: The extract and platycodin-D from PG root have a quick sperm-killing effect in a short time in vitro by disrupting the integrity of sperm membrane (main head).
Asunto(s)
Extractos Vegetales/farmacología , Platycodon/química , Saponinas/farmacología , Espermicidas/farmacología , Espermatozoides/efectos de los fármacos , Triterpenos/farmacología , Adulto , Membrana Celular/efectos de los fármacos , Humanos , MasculinoRESUMEN
OBJECTIVE: To compare sperm chromatin dispersion (SCD) test with terminal deoxynucleotidyle transferase-mediated terminal uridine nick-end labeling (TUNEL) assay in assessing DNA fragmentation in human sperm through bright-field microscopy. DESIGN: Prospectively designed, side-by-side comparative study. SETTING: Medical genetics laboratory in a provincial research institution. PATIENT(S): Sixty male patients presented for infertility evaluation and 30 fertile, volunteer sperm donors. INTERVENTION(S): Semen analysis, SCD test, and TUNEL assay on the same semen sample and on the same spermatozoa. MAIN OUTCOME MEASURE(S): Sperm DNA fragmentation, determined by SCD test score or TUNEL assay score. RESULT(S): Sperm chromatin dispersion test and TUNEL assay identified similar proportions of sperm cells with DNA fragmentation in the same semen samples. When the SCD test and TUNEL assay were performed simultaneously on the same spermatozoa, TUNEL-negative sperm showed a large halo, whereas TUNEL-positive sperm showed no halo after Diff-Quik staining. However, in some sperm cells, DNA damage was detected by the SCD test but not by the TUNEL assay. CONCLUSION(S): Sperm chromatin dispersion test and TUNEL assay are both effective in detecting sperm DNA damage. Using bright-field microscopy, the SCD test appears to be more sensitive than the TUNEL assay.
Asunto(s)
Ensamble y Desensamble de Cromatina , Fragmentación del ADN , Etiquetado Corte-Fin in Situ/métodos , Microscopía/métodos , Espermatozoides/ultraestructura , Adulto , Forma de la Célula , Ensamble y Desensamble de Cromatina/fisiología , Fertilidad/genética , Fertilidad/fisiología , Humanos , Infertilidad Masculina/genética , Infertilidad Masculina/metabolismo , Masculino , Análisis de Semen/métodos , Espermatozoides/citología , Espermatozoides/metabolismoRESUMEN
OBJECTIVE: To compare the effects of sperm chromatin dispersion (SCD) test and TdT-mediated dUTP nick end labeling (TUNEL) assay in assessing the DNA fragmentation in human sperm. METHODS: Motile sperms were isolated from the semen samples obtained from 20 healthy fertile men and 32 clinically infertile patients by swim-up technique, and underwent SCD and TUNEL to analyze the DNA fragmentation. RESULTS: The rate of sperm with DNA damage of the infertile patients was 12.8% +/- 5.8% tested by SCD, significantly higher than that of the healthy fertile men (7.6% +/- 3.3%, t = 3.576, P = 0.001), and the rate of sperm with DNA damage of the infertile patients was 11.1% +/- 5.1% tested by TUNEL assay, significantly higher than that of the healthy fertile men (6.8% +/- 2.8%, t = 3.467, P = 0.001). The proportion of sperm cell with abnormal DNA integrity measured by SCD test was correlated strongly with that determined by TUNEL for the infertile men (r = 0.841, P = 0.000) and for the fertile men too (r = 0.823, P = 0.000). The rate of sperm with DNA damage measured by SCD were not significantly different from those of TUNEL-positive sperm in fertile men (t = 1.996, P = 0.060). The rate of sperm with DNA damage measured by SCD was significantly higher than that measured by TUNEL among infertile patients (t = 3.023, P = 0.005). CONCLUSION: The presence of sperm DNA damage may lead to male infertility. SCD is simpler, cheaper and more reliable than TUNEL in testing the sperm DNA damage.
Asunto(s)
Daño del ADN , ADN/genética , Infertilidad Masculina/diagnóstico , Infertilidad Masculina/genética , Espermatozoides/química , Adulto , Estudios de Casos y Controles , Fragmentación del ADN , Humanos , Infertilidad Masculina/metabolismo , Masculino , Motilidad EspermáticaRESUMEN
OBJECTIVE: To explore the effectiveness of percutaneous vasal sperm aspiration (PVSA) in combination with intrauterine insemination (IUI) to treat infertile men with anejaculation. DESIGN: Clinical study. SETTING: Department of reproductive endocrinology and andrology of a family planning research clinic. PATIENT(S): Twenty-six anejaculatory infertile men. INTERVENTION(S): Spermatozoa obtained from the vas deferens by percutaneous aspiration were incubated in sperm preparation medium. MAIN OUTCOME MEASURE(S): Sperm quality by PVSA and pregnancy outcome. RESULT(S): Thirty-four PVSA-IUI procedures were performed in 26 men with anejaculation. Nineteen pregnancies were achieved (pregnancy rate, 73.1%). Mean (+/-SD) values for sperm variables were as follows: motility, 78.6% +/- 14.2%; progressive motility, 60.4% +/- 11.2%; density, 37.6 +/- 13.2 x 10(6) cells/mL; total count, 35.2 +/- 13.2 x 10(6) cells; and abnormal sperm, 18.6% +/- 7.6%. CONCLUSION(S): Percutaneous vasal sperm aspiration may obtain high-motility sperm, and PVSA plus IUI is an effective treatment for male infertility with anejaculation.