ObjectiveTo investigate the effects of Zhibai Dihuang Decoction (ZDD) on sperm mitochondrial permeability transition (MPT) in the rat model of ureaplasma urealyticum (UU) infection (UUI).

METHODSNinety male SD rats were randomly divide into five groups: normal control, UUI model control, ZDD, doxycycline, and ZDD + doxycycline. The UUI model was established in the latter four groups of rats by UU injection into the bladder. On the second day after modeling, the animals of the normal control and UUI model control groups were treated intragastrically with 0.9% sodium chloride solution and those in the other groups with corresponding drugs, all for 21 consecutive days. At 24 hours after drug withdrawal, epididymal samples were obtained for detection of the protein and mRNA expressions of VDAC2 and ANT4 in the sperm mitochondria by RT-PCR and Western blot respectively and determination of the contents of adenosine monophosphate (AMP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) and energy charge (EC) in the sperm mitochondria by high-performance liquid chromatography.

RESULTSThe protein expressions of VDAC2 and ANT4 in the rat sperm mitochondria were 0.626 ± 0.074 and 0.527 ± 0.096 in the normal control group, 0.039 ± 0.011 and 0.044 ± 0.011 in the UUI model control group, 0.101 ± 0.037 and 0.127 ± 0.040 in the ZDD group, 0.236 ± 0.070 and 0.253 ± 0.054 in the doxycycline group, and 0.475 ± 0.064 and 0.367 ± 0.086 in the ZDD + doxycycline group, significantly lower in the UUI model control than in the normal control group (P<0.05 and P<0.01), but remarkably higher in the doxycycline and ZDD + doxycycline groups than in the UUI model control (P<0.01) and the ZDD group (P<0.05 and P<0.01), and the expression of VDAC2 was markedly higher in the ZDD + doxycycline than in the doxycycline group (P<0.01). The mRNA expressions of VDAC2 and ANT4 were 0.008 ± 0.001 035 and 0.026 50 ± 0.003 401 in the normal control group, 0.000 79 ± 0.000 226 and 0.001 64 ± 0.000 205 in the UUI model controls, 0.002 06 ± 0.000 861 and 0.005 04 ± 0.002 537 in the ZDD group, 0.003 34 ± 0.000 229 and 0.008 57 ± 0. 000 690 in the doxycycline group, and 0.004 85 ± 0.000 495 and 0.013 13 ± 0.000 826 in the ZDD + doxycycline group, significantly lower in the UUI model control than in the normal control group (P<0.05 and P<0.01), but remarkably higher in the ZDD, doxycycline and ZDD + doxycycline groups than in the UUI model controls (P<0.01) as well as in the doxycycline and ZDD + doxycycline groups than in the ZDD group (P<0.01) and in the ZDD + doxycycline than in the doxycycline group (P<0.01). The levels of ATP, ADP, AMP and EC in the sperm mitochondria were (203.41 ± 13.16) mg/L, (129.87 ± 14.68) mg/L, (149.05 ± 5.65) mg/L and 0.56 ± 0.01 in the normal control group, (96.22 ± 12.55) mg/L, (99.87 ± 3.28) mg/L, (212.53 ± 19. 43) mg/L and 0.36 ± 0.03 in the UUI model control group, (101.99 ± 5.97) mg/L, (104.99 ± 16.40) mg/L, (183.97 ± 12.43) mg/L and 0.40 ± 0.01 in the ZDD group, (159.44 ± 33.16) mg/L, (118.51 ± 12.99) mg/L, (160.64 ± 14.19) mg/L and 0.50 ± 0.06 in the doxycycline group, and (194.07 ± 9.36) mg/L, (121.62 ± 9.41) mg/L, (150.21 ± 12.87) mg/L and 0.55 ± 0.01 in the ZDD + doxycycline group. The levels of ATP and EC were significantly lower and that of AMP higher in the UUI model control than in the normal control group (P<0.01), while the former two were remarkably higher and the latter one lower in the doxycycline and ZDD + doxycycline groups than in the UUI model controls (P<0.05 and P<0.01). Compared with the ZDD + doxycycline group, the ZDD group showed significantly decreased ATP and EC but increased AMP, while the doxycycline group exhibited decreases in both ATP and EC (P<0.05 and P<0.01).

CONCLUSIONSZDD can upregulate the decreased protein and mRNA expressions of VDAC2 and ANT4 in the sperm mitochondria and improve sperm mitochondrial permeability transition and mitochondrial energy metabolism in rats with UU infection, which may be one of its action mechanisms in the treatment of UU infection-induced male infertility.

Animals ; Anti-Bacterial Agents ; therapeutic use ; Doxycycline ; therapeutic use ; Drugs, Chinese Herbal ; metabolism ; therapeutic use ; Energy Metabolism ; Epididymis ; Humans ; Infertility, Male ; Male ; Mitochondria ; drug effects ; Permeability ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spermatozoa ; drug effects ; Ureaplasma Infections ; drug therapy ; Ureaplasma urealyticum ; Voltage-Dependent Anion Channel 2 ; metabolism

OBJECTIVE: The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipid-associated membrane proteins (LAMPs) in the host innate immune system, specifically their effect on Toll-like receptors (TLRs). METHODS: LAMPs were derived from U. urealyticum strains, and human amniotic epithelial cells (HAECs) were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay (ELISA) and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry. RESULTS: LAMPs induced HAECs to produce inflammatory cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor (anti-hTLR2-IgA). CONCLUSIONS LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.
Amnion/cytology* ; Amniotic Fluid/cytology* ; Cytokines/metabolism* ; Dose-Response Relationship, Drug ; Epithelial Cells/metabolism* ; Female ; Humans ; Inflammation ; Interleukin-6/metabolism* ; Interleukin-8/metabolism* ; Lipids/chemistry* ; Lipopolysaccharides/metabolism* ; Membrane Proteins/metabolism* ; Placenta/metabolism* ; Pregnancy ; Toll-Like Receptor 2/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Up-Regulation ; Ureaplasma urealyticum/metabolism*

Objective: To explore the effects of Zhibai Dihuang Decoction (ZDD) on mitochondrial cytochrome oxidase (COX) in the spermatogenic cells of rats with ureaplasma urealyticum (UU) infection. METHODS: From forty 4－5 months old SD rats, 30 were randomly selected for the establishment of the model of testicular UU infection by inoculating the bladder with UU suspension and the other 10 injected with normal saline as controls (group A). At 7 days after inoculation, the rat models of testicular UU infection were treated orally with normal saline (group B), ZDD at 1 g per kg of the body weight per day (group C), and azithromycin at 0.105 g per kg of the body weight per day (group D), respectively, once daily for 21 days. Then all the animals were sacrificed and the epididymal and testicular tissues collected for examination of sperm motility with the color sperm dynamic detection system, measurement of the COX activity with the immunohistochemical DAB method, and determination of the mRNA expressions of COXⅠ and COXⅡ by RT-PCR. RESULTS: Compared with group A, group B showed significant decreases in such sperm parameters as grade a sperm (［1.03 ± 0.09］ vs ［0.07 ± 0.03］ %, P<0.01), grade b sperm (［2.07 ± 0.52］ vs ［0.35 ± 0.13］ %, P<0.01), straight line velocity (VSL) (［10.95 ± 0.98］ vs ［6.78 ± 1.05］ μm/s, P<0.01), curvilinear velocity (VCL) (［42.03 ± 1.35］ vs ［38.10 ± 7.65］ μm/s, P>0.05), average path velocity (VAP) (［16.22 ± 1.52］ vs ［10.05 ± 1.80］ μm/s, P<0.01), and the mRNA expressions of COX Ⅰ (［2.25 ± 0.24］ vs ［0.93 ± 0.10］ %, P<0.01) and Ⅱ (［6.72 ± 0.37］ vs ［2.95 ± 0.78］ %, P<0.01). After treatment, all the parameters were remarkably increased in groups C and D (grade a sperm: ［1.11 ± 0.30］ and ［0.60 ± 0.19］%; grade b sperm: ［2.40 ± 0.59］ and ［1.32 ± 0.27］ %; VSL: ［12.11 ± 1.62］ and ［11.47 ± 1.21］ μm/s; VCL: ［54.30 ± 2.35］ and ［45.75 ± 1.64］ μm/s; VAP ［18.40 ± 1.27］ and ［16.69 ± 1.02］ μm/s; expression of COXⅠ mRNA: ［1.86 ± 0.30］ and ［1.74 ± 0.17］ %) as compared with those in group B (P<0.05or P<0.01) except the COX activity and the expression of COX Ⅱ mRNA (P>0.05), and all the parameters were significantly higher in group C than in D (P<0.05or P<0.01). CONCLUSIONS UU infection can reduce grades a and b sperm, linear, curvilinear and mean sperm velocities, and the mRNA expressions of COX Ⅰ and Ⅱ while ZDD can improve these parameters. The improvement of sperm motility may not be associated with the activity of COX, and the COX activity may be related to the mRNA expression of COX II but not that of COXⅠ.
Animals ; Anti-Bacterial Agents ; therapeutic use ; Azithromycin ; therapeutic use ; Drugs, Chinese Herbal ; pharmacology ; Electron Transport Complex IV ; metabolism ; Epididymis ; drug effects ; enzymology ; Humans ; Male ; Mitochondria ; drug effects ; enzymology ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; Spermatozoa ; drug effects ; enzymology ; physiology ; Ureaplasma Infections ; drug therapy ; enzymology ; Ureaplasma urealyticum

OBJECTIVETo observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD.

METHODSTotally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot.

RESULTSThe UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01).

CONCLUSIONZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

Animals ; Calcium Channels ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Infertility ; Male ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Semen Analysis ; Sperm Motility ; Spermatozoa ; TRPV Cation Channels ; metabolism ; Testis ; Ureaplasma Infections ; Ureaplasma urealyticum

OBJECTIVETo study the effect of Zhibai Dihuang Pill (ZBDHP) on urokinase-type plasminogen activator (uPA) and sperm quality in ureaplasma urealyticum (Uu) infection infertile patients.

METHODSRecruited were 80 infertility patients with Uu infection at Andriatrics Clinics and Department of Reproduction, including 130 cases of positive Uu semen and 50 cases of negative Uu semen. Patients with positive Uu semen were randomly assigned to the observation group (72 cases) and the control group (58 cases) according to the visit sequence. All patients took antibiotics for 2 weeks. Patients in the observation group additionally took ZBDHP, 6 g each time, twice daily. Those in the control group additionally took Vit E (100 mg each time, twice per day) and ATP (40 mg each time, twice per day). The therapeutic course for all was 90 days. Semen parameters and uPA contents of the sperm membrane were detected and comparatively analyzed.

RESULTSThe sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm in Uu positive infected patients were lower than those in Uu negative infected patients with statistical difference (P < 0.05), but with no significant difference in the sperm density between the two groups (P > 0.05). There was no statistical difference in pre-treatment sperm membrane uPA contents and sperm parameters between the two groups (P > 0.05). Compared with before treatment in the same group, the sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm obviously increased in the two groups with statistical difference (P < 0.05). After treatment, the sperm membrane uPA content increased more obviously in the observation group, with statistical difference when compared with the control group (P < 0.05).

CONCLUSIONSInfection with Uu leads to decreased uPA content of sperm membrance and the sperm motility. ZBDHP could effectively treat Uu infected infertility possibly through fighting against Uu damaged sperm membrane and make the sperm membrane uPA content return to normal, and elevate the fertilizability of sperms.

Anti-Bacterial Agents ; administration & dosage ; pharmacology ; therapeutic use ; Communicable Diseases ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Humans ; Infertility ; Infertility, Male ; Male ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; Ureaplasma Infections ; drug therapy ; Ureaplasma urealyticum ; drug effects ; Urokinase-Type Plasminogen Activator ; metabolism

The aim of this study was to determine whether intra-amniotic infection/inflammation (IAI) was associated with subsequent ruptured membranes in women with preterm labor and intact membranes who had a clinically indicated amniocentesis. This retrospective cohort study included 237 consecutive women with preterm labor (20-34.6 weeks) who underwent amniocentesis. The clinical and laboratory parameters evaluated included demographic variables, gestational age, C-reactive protein (CRP) and amniotic fluid (AF) white blood cell, interleukin-6 (IL-6) and culture results. IAI was defined as a positive AF culture and/or an elevated AF IL-6 level (>2.6 ng/mL). The primary outcome was ruptured membranes in the absence of active labor occurring within 48 hours of amniocentesis. Preterm premature rupture of membranes subsequently developed in 10 (4.2%) women within 48 hr of amniocentesis. Multivariate analysis demonstrated that only IAI was independently associated with the ruptured membranes occurring within 48 hr of amniocentesis. In the predictive model based on variables assessed before amniocentesis, only CRP level was retained. IAI is an independent risk factor for subsequent ruptured membranes after clinically indicated amniocentesis in preterm labor. Prior to amniocentesis, measurement of serum CRP level can provide a risk assessment for the subsequent development of ruptured membranes after the procedure.
Adult ; Amniocentesis/*adverse effects ; Amnion/physiopathology ; Amniotic Fluid/cytology/metabolism/microbiology ; Bacterial Infections/*etiology/microbiology ; C-Reactive Protein/analysis ; Cohort Studies ; Demography ; Female ; Gestational Age ; Humans ; Inflammation/*etiology ; Interleukin-6/metabolism ; Leukocytes/cytology ; Multivariate Analysis ; Mycoplasma/isolation & purification ; Obstetric Labor, Premature/*etiology ; Pregnancy ; ROC Curve ; Retrospective Studies ; Risk Factors ; Ureaplasma urealyticum/isolation & purification

OBJECTIVETo determine the impact of Ureaplasma urealyticum (Uu) infection on the integrity of sperm plasma membrane in infertile males.

METHODSSixty-three semen samples were divided into a Uu infection group (n = 32) and a normal control group (n = 31). Conventional semen analyses were performed by computer-assisted semen analysis (CASA) and Uu detected by the culture method. The semen samples were washed with PBS and dyed by SYBR-14/PI double fluorescent staining, followed by detection of the integrity of sperm plasma membrane by flow cytometry. The percentage of the sperm with intact plasma membrane was indicated as the percentage of sperm emitting green fluorescence (SYBR-14+/PI-%).

RESULTSThe Uu infection group showed a significantly decreased integrity of sperm plasma membrane ([45.14 +/- 10.69]%) and reduced percentage of grade a + b sperm ([23.29 +/- 8.81]%) as compared with the normal control group ([72.68 +/- 9.91]% and [46.32 +/- 9.54]%) (P < 0.01). But there were no significant differences in the semen volume, pH value, and sperm concentration between the two groups (P > 0.05).

CONCLUSIONUu infection decreases the integrity of sperm plasma membrane, which might be an important factor of male infertility.

Adult ; Case-Control Studies ; Cell Membrane ; pathology ; Flow Cytometry ; Humans ; Infertility, Male ; microbiology ; pathology ; physiopathology ; Male ; Organic Chemicals ; Semen Analysis ; methods ; Spermatozoa ; metabolism ; pathology ; Ureaplasma Infections ; pathology ; physiopathology ; Ureaplasma urealyticum ; Young Adult

OBJECTIVETo observe the effects of Zhibai Dihuang Decotion (ZDD) on the ureaplasma urealyticum (UU)-infected rats' spermatogenic cell apoptosis and expressions of Caspase-3 and Caspase-9.

METHODS45 out of 60 male SD rats were randomly selected and made into the UU infected animal model. The rest 15 were taken as the sham-operation group. The UU infected model animals were then randomly divided into the model group, the minocycline group, and the ZDD group. From the 10th day after inoculation, normal saline was given to rats of the model group and the sham-operation group by gastrogavage, while corresponding medicines were given to rats in the minocycline group and the ZDD group. All rats were killed after 21 successive days of gastrogavage. The apoptosis rate of reproductive cells, Caspase-3 and Caspase-9 expression levels and ultrastructure changes of spermatogenic cells of each group were detected and compared.

RESULTSThere was statistical difference in the positive rate of the UU cultivation results, the apoptosis rate of reproductive cells, Caspase-3 and Caspase-9 expression levels in the sham-operation group, the minocycline group, and the ZDD group when compared with the model group (P<0.05). There was statistical difference in the aforesaid indices in the minocycline group and the ZDD group when compared with the sham-operation group (P<0.05). Still there was no statistical difference in the aforesaid indices between the minocycline group and the ZDD group (P>0.05).

CONCLUSIONSUU infection can lead to the increasing of spermatogenic cell's apoptosis in rats. ZDD could actively inhibit the growth and production of UU with anti-UU. One of the mechanisms of ZDD in treating UU infection and improving the sperm quality is through regulating the expressions of the apoptosis effect factors Caspase-3 and Caspase-9.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Spermatozoa ; cytology ; metabolism ; Ureaplasma Infections ; drug therapy ; metabolism ; pathology ; Ureaplasma urealyticum

OBJECTIVETo understand Ureaplasma urealyticum (Uu) infection, analyzed the influence of Uu infection on the seminal quality and the accessory genetical gland function in male infertility patients, and investigate its mechanism.

METHODSWe cultured 202 semen samples collected from male infertility patients and analyzed the influence of Uu infection on seminal parameters and the biochemical indexes of the seminal plasma.

RESULTSThe Uu infection rate was 33.7% in the infertile males, with no statistic differences between the Uu positive and negative groups either in the average age (28.9 +/- 4.7 yrs vs 29.6 +/- 4.0 yrs, P = 0.250) or in the seminal quantity (2.93 +/- 1.32 ml vs 2.86 +/- 1.52 ml, P = 0.774). The sperm density, motility and vitality were (84.37 +/- 52.92) x 10(6) ml, (44.62 +/-22.13) % and (38.40 +/- 15.61) % in the Uu positive group, significantly lower than (101.90 +/- 43.90) x 10(6) ml, (51.83 +/- 19.88) % and (44.45 +/- 15.47) % in the Uu negative group (P = 0.025, P = 0.036 and P = 0.020). The seminal pH value was normal in both of the groups, but significantly higher in the Uu positive than in the negative group (7.32 +/- 0.10 vs 7.19 +/- 0.29, P = 0.003). VCL, VSL, VAP and MAD were significantly lower, while BCF was significant higher in the former than in the latter [(33.97 +/- 8.96) microm/s vs (39.70 +/- 8.14) microm/s, t = 4.113, P < 0.001; (22.29 +/- 6.06) microm/s vs (25.20 +/- 6.67) microm/s, t = 2.684, P = 0.008; (25.96 +/- 6.83) microm/s vs (30.02 +/- 6.81) microm/s, t = 3.537, P < 0.001; 46.60 +/- 13.68 vs 54.23 +/- 15.14, t = 3.112, P = 0.002; (6.12 +/- 1.89) Hz vs (5.22 +/- 1.64) Hz, t = 3. 164, P = 0.002]. All the five indexes were influenced by Uu infection. Compared with the negative group, the seminal plasma alpha-glucosidase was significantly decreased in the positive group [(40.0 +/-18.7) U/ml vs (47.9 +/- 21.0) U/ml, t = 2.248, P = 0.026], and the risk of the decrease was 2.12 times higher. No statistic difference was observed in seminal plasma acid phosphatase and seminal plasma fructose between the two groups.

CONCLUSIONUu infection in the genital tract is an important factor of seminal quality reduction in infertile men and may cause a decreased secretion of alpha-glucosidase in the epididymis, but it hardly influences the prostate and seminal vesicle.

Adult ; Genital Diseases, Male ; microbiology ; physiopathology ; Humans ; Infertility, Male ; physiopathology ; Male ; Middle Aged ; Semen ; cytology ; metabolism ; microbiology ; Sperm Count ; Sperm Motility ; Ureaplasma Infections ; microbiology ; physiopathology ; Ureaplasma urealyticum ; isolation & purification ; alpha-Glucosidases ; metabolism

OBJECTIVETo study the influence of Ureaplasma urealyticum (Uu) infection on the sperm-egg binding associated molecule, sulfogalactosylglycerolipid (SGG).

METHODSEpididymal sperm was collected from adult mice. The sperm suspension was randomly divided into 4 groups: Uu group (coincubated with Uu suspension), medium group (coincubated with Uu medium), normal group and PRS group. The indirect immunofluorescence technique was used to localize SGG on the sperm membrane and to observe the influence of Uu on SGG.

RESULTSIn the epididymal sperm, SGG was localized to the head plasma membrane overlaying the acrosomal region. The SGG-positive rate of the sperm coincubated with Uu medium was 82.0%, while that of the sperm coincubated with Uu suspension was reduced to 39.0% (P = 0.001).

CONCLUSIONUu can adhere to the sperm surface. SGG might be a membrane receptor on the sperm surface for Uu infection of the mammalian male genital tract. The blockage of SGG by Uu might be one of the molecular mechanisms correlative to male infertility induced by Uu infection.

Animals ; Cell Membrane ; metabolism ; Galactolipids ; biosynthesis ; Infertility, Male ; etiology ; Male ; Mice ; Mice, Inbred BALB C ; Random Allocation ; Sperm-Ovum Interactions ; physiology ; Spermatozoa ; metabolism ; Ureaplasma Infections ; complications ; metabolism ; Ureaplasma urealyticum