ObjectiveTo investigate the effects of resveratrol in the cryopreservation medium on the quality and function of post-thaw sperm.

METHODSSemen samples were obtained from 50 normozoospermic and 50 oligoasthenozoospermic men, liquefied and then cryopreserved in the glycerol-egg yolk-citrate (GEYC) medium with or without 30 μmol/L resveratrol. Sperm motility, viability and acrosome reaction (AR) were examined before and after thawing. Sperm lipid peroxidation and the level of reactive oxygen species (ROS) were measured using commercial malondialdehyde (MDA) and the ROS assay kit. Sperm mitochondrial membrane potential (MMP) and DNA damage were determined by Rhodamine 123 staining and TUNEL.

RESULTSThe percentage of progressively motile sperm (PMS), total sperm motility, sperm viability, MMP and AR were significantly decreased (P <0.05) while the levels of sperm ROS, MDA and DNA fragmentation index (DFI) remarkably increased in both the normozoospermia and oligoasthenozoospermia groups after cryopreservation as compared with those in the fresh ejaculate (P <0.05). In comparison with the non-resveratrol control, the post-thaw sperm cryopreserved with 30 μmol/L resveratrol showed markedly higher PMS (［32.7 ± 4.8］ vs ［43.1 ± 6.3］ %, P <0.05), total motility (［44.8 ± 6.9］ vs ［56.9 ± 7.4］ %, P <0.05), viability (［52.3 ± 6.1］ vs ［67.5 ± 5.6］ %, P <0.05), MMP (［56.5 ± 7.0］ vs ［63.4 ± 7.5］ %, P <0.05) and AR (［16.6 ± 3.8］ vs ［26.3 ± 4.7］ %, P <0.05) but lower ROS, MDA and DFI (all P <0.05) in the normozoospermia group, and so did the post-thaw sperm in the oligoasthenozoospermia group, with a particularly lower DFI (［28.5 ± 4.8］ vs ［36.3 ± 5.7］%, P <0.01).

CONCLUSIONSResveratrol in the cryopreservation medium can improve the quality and function of post-thaw human sperm by reducing cryopreservation-induced sperm injury and the level of ROS.

Acrosome ; drug effects ; Animals ; Antioxidants ; pharmacology ; Cryopreservation ; methods ; DNA Fragmentation ; Humans ; Lipid Peroxidation ; Male ; Malondialdehyde ; Membrane Potential, Mitochondrial ; Reactive Oxygen Species ; analysis ; Resveratrol ; pharmacology ; Semen Analysis ; Semen Preservation ; adverse effects ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects ; physiology

ObjectiveTo investigate the effects of Qiangjing Tablets (QJT) on sperm quality and the MAPK signaling pathway in the SD rat model of asthenospermia (AS).

METHODSA total of 100 male SD rats were randomly divided into five groups of equal number, blank control, AS model control, high-dose QJT, medium-dose QJT, and low-dose QJT. All the rats were intragastrically administered ORN at 200 mg/kg/d for establishment of the AS model except those in the blank control group, which were given 1% CMC sodium solution at 1 ml/100 g by gavage. Meanwhile the animals of the high-, medium-, and low-dose QJT groups were gavaged with QJT at 6700, 3300 and 1700 mg/kg/d, respectively, qd 6 days a week for 20 days. Then the testis issue and the apoptosis of the testicular cells were observed under the electron microscope, the expression of vimentin in the testis was determined with the immunohistochemical SP method, that of ERK1/2 detected by Western blot, and the concentration of TGF-β1 in the semen measured by ELISA.

RESULTSThe AS model controls showed round nuclei of spermatocytes, homogeneously distributed chromatins, broken or lost mitochondria, and expanded rough endoplasmic reticulum in the testis tissue. In comparison, the rats of the high-, medium-, and low-dose QJT groups exhibited round nuclei of spermatocytes, homogeneously distributed chromatins, and well-structured mitochondria, rough endoplasmic reticulum and ribosome, which were all similar those of the blank controls. Compared with the blank controls, the AS model rats manifested significantly increased expressions of ERK1/2 (1.00 ± 0.00 vs 1.26 ± 0.10, P<0.01) and vimentin (0.16 ± 0.01 vs 0.17 ± 0.01, P<0.01) and apoptosis rate of cells in the testis tissue (［9.20 ± 3.07］ vs ［42.20 ± 9.17］ %, P<0.01), but decreased level of TGF-β1 in the semen (［627.67 ± 26.07］ vs ［566.73 ± 68.44］ ng/ml, P<0.05). In comparison with the model controls, the rats of the high- and medium- -dose QJT groups presented remarkably down-regulated expressions of ERK1/2 (1.26 ± 0.10 vs 1.14 ± 0.08, P<0.01; 1.26 ± 0.10 vs 1.18 ± 0.05, P<0.05) and vimentin (0.17 ± 0.01 vs 0.16 ± 0.01, P<0.01; 0.17 ± 0.01 vs 0.17 ± 0.09, P<0.05) and decreased rate of cell apoptosis (［42.20 ± 9.17］ vs ［21.60 ± 5.94］ %, P<0.01; ［42.20 ± 9.17］ vs ［33.95 ± 6.39］ %, P<0.05). The concentration of TGF-β1 in the semen was markedly lower in the high-dose QJT than in the AS model control group (［621.78 ± 30.80］ vs ［566.73 ± 68.44］ ng/ml, P < 0.05).

CONCLUSIONSQiangjing Tablets could improve semen quality in asthenospermia rats by acting against oxidative stress.

Animals ; Apoptosis ; Asthenozoospermia ; enzymology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Semen ; Semen Analysis ; Signal Transduction ; Spermatozoa ; Testis ; metabolism ; ultrastructure ; Transforming Growth Factor beta1 ; metabolism ; Vimentin ; metabolism

OBJECTIVE: To explore the inhibitory effect of PSB0739 on the formation of semen-derived amyloid fibrils. METHODS: PAP248-286 (440 μmol/L) was incubated with PSB0739 at different concentrations, and at different time points of incubation, aliquots were taken from each sample for Congo red staining to detect the formation of amyloid fibers. The morphology of amyloid fibrils incubated in the presence or absence of PSB0739 was visualized using transmission electron microscope. The effect of PSB0739 on amyloid fibril formation was determined using virus infection assays at different time points, and the surface charges of amyloid fibril incubated with PSB0739 were calculated using a Zeta potentiometer. The cytotoxicity of PSB0739 in Hela cells was determined using MTT assay. The antiviral effect of PSB0738 against HIV- 1 was evaluated by infection assay. RESULTS: PSB0739 inhibited SEVI fibril formation in a dose-dependent manner . At 48 h of incubation, 220 μmol/L of PSB0739 obviously inhibited the formation of amyloid fibrils in 440 μmol/L of SEVI. Transmission electron microscopy revealed that 220 μmol/L PSB0739 prevented PAP248- 286 (440 μmol/L) from forming amyloid fibrils. PSB0739 antagonized SEVI-mediated enhancement of HIV-1 infection, and 1760 μmol/L of PSB0739 completely reversed the positive charge of SEVI ( < 0.05). PSB0739 below the concentration of 62.5 μmol/L showed no obvious cytotoxicity in Hela cells (>0.05). PSB0739 showed a direct anti-HIV activity with an IC of 21.77±5.15 μmol/L. CONCLUSIONS PSB0739 can inhibit the formation of semen-derived amyloid fibrils .
Amyloid ; chemistry ; drug effects ; Anti-HIV Agents ; pharmacology ; Dose-Response Relationship, Drug ; HIV Infections ; drug therapy ; HIV-1 ; drug effects ; HeLa Cells ; Humans ; In Vitro Techniques ; Microscopy, Electron, Transmission ; Purinergic P2Y Receptor Antagonists ; pharmacology ; Semen ; chemistry

OBJECTIVE: To determine the mitigating effects of sodium 4-phenylbutyrate (4-PBA) on high-fat diet (HFD)-induced spermatogenesis dysfunction. METHODS: Male rats (n = 30) were randomly divided into three groups: control, HFD, and 4-PBA (HFD +4-PBA). After 13 weeks, rats were euthanized. Testes and epididymis were harvested for further analysis. Sex hormones were detected, and hematoxylin and eosin staining was performed to examine the histological changes in the testes. Semen samples were collected to evaluate sperm quality. Spermatogenic cell apoptosis was detected by TUNEL assay. RESULTS: Compared with the control group, the final body weight and body weight gain were significantly higher in HFD-fed rats, while the testicle/body weight ratios were lower (P < 0.05). In HFD-fed rats, obvious pathological changes in the testicular tissue were observed. Treatment with 4-PBA attenuated HFD-induced histological damage, ameliorated the HFD-induced decrease in serum testosterone (T), and reduced the rate of testicular cell apoptosis (P < 0.05) in obese male rats. Finally, 4-PBA significantly improved semen parameters in HFD rats (P < 0.05). CONCLUSION HFD exposure induced detrimental effects on spermatogenesis, semen quality, serum T level, and testicular cell apoptosis in rats. Treatment with 4-PBA ameliorated HFD？induced impaired spermatogenesis via inhibition of apop-tosis in rats. 4-PBA may have therapeutic value in the treatment of obesity？related impairment of spermatogenesis.
Animals ; Diet, High-Fat ; adverse effects ; Male ; Phenylbutyrates ; pharmacology ; Rats, Sprague-Dawley ; Semen Analysis ; Spermatogenesis ; drug effects ; Testis ; drug effects ; pathology ; Testosterone ; blood

Proton-pump inhibitors (PPIs) are among the most widely used drugs worldwide. PPI use has recently been linked to adverse changes in semen quality in healthy men; however, the effects of PPI use on semen parameters remain largely unknown specifically in cases with male factor infertility. We examined whether PPI use was associated with detrimental effects on semen parameters in a large population of subfertile men. We retrospectively reviewed data from 12 257 subfertile men who had visited our fertility clinic from 2003 to 2013. Patients who reported using any PPIs for >3 months before semen sample collection were included; 7698 subfertile men taking no medication served as controls. Data were gathered on patient age, medication use, and conventional semen parameters; patients taking any known spermatotoxic medication were excluded. Linear mixed-effect regression models were used to test the effect of PPI use on semen parameters adjusting for age. A total of 248 patients (258 samples) used PPIs for at least 3 months before semen collection. In regression models, PPI use (either as the only medication or when used in combination with other nonspermatotoxic medications) was not associated with statistically significant changes in semen parameters. To our knowledge, this is the largest study to compare PPI use with semen parameters in subfertile men. Using PPIs was not associated with detrimental effects on semen quality in this retrospective study.
Adult ; Case-Control Studies ; Humans ; Infertility, Male/epidemiology* ; Male ; Proton Pump Inhibitors/therapeutic use* ; Retrospective Studies ; Semen Analysis ; Sperm Count ; Sperm Motility/drug effects* ; Spermatozoa/drug effects*

Objective: To investigate the clinical therapeutic effects of Yijingfang, a Chinese medicinal liquid, on asthenospermia. METHODS: We randomly divided 450 asthenospermia patients into a treatment group (n = 300) and a control group (n = 150), the former treated with Yijingfang once half a dose, bid, and the latter with Wuziyanzong Pills (9 g, bid) + L-carnitine oral liquid (10 ml, bid), both for 3 months. Before and at 1, 2, and 3 months after medication, we compared the semen volume, sperm concentration, percentages of progressively motile sperm (PMS) and total motile sperm (TMS), and semen liquefaction time between the two groups of patients. RESULTS: No statistically significant difference was observed in the semen parameters between the treatment and control groups before medication (P >0.05). In comparison with the baseline, the treatment group showed significant differences at 1, 2, and 3 months after medication in sperm concentration (［35.96 ± 8.50］ vs ［49.66 ± 10.91］, ［55.21 ± 11.46］, ［74.90 ± 13.07］ ×10⁶/ml, P <0.01), PMS (［19.72 ± 2.06］ vs ［23.81 ± 2.56］, ［26.12 ± 2.34］, and ［32.17 ± 1.62］ %, P <0.01) and TMS (［28.86 ± 2.70］ vs ［34.17 ± 3.43］, ［36.59 ± 3.36］, and ［47.08 ± 2.97］ %, P <0.01), but not in the semen volume (［3.35 ± 0.99］ vs ［3.15 ± 1.06］, ［3.12 ± 0.90］, and ［3.27 ± 0.78］ ml, P >0.05) or semen liquefaction time (［32.31 ± 8.15］ vs ［31.68 ± 3.14］, ［30.38 ± 3.44］, and ［30.86 ± 2.42］ min, P >0.05); the control group exhibited similar results at the three time points in sperm concentration (［36.85 ± 6.88］ vs ［40.53 ± 8.32］, ［47.51 ± 12.73］, and ［56.14 ± 11.98］ ×10⁶/ml, P <0.01), PMS (［20.26 ± 2.73］ vs ［25.17 ± 2.64］, ［27.23 ± 2.25］, and ［31.89±2.27］ %, P <0.01), and TMS (［30.03 ± 2.67］ vs ［33.89±2.26］, ［37.38±4.79］, and ［40.35±3.06］ %, P <0.01), but not in the semen volume (［3.03 ± 1.09］ vs ［3.16±1.78］, ［3.15±0.96］, and ［3.12±0.65］ ml, P >0.05) or semen liquefaction time (［30.25 ± 5.20］ vs ［29.36±4.25］, ［28.21±3.26］, and ［28.33±3.59］ min, P >0.05). There were statistically significant differences between the treatment and control groups in the increase rates of sperm concentration and TMS after medication (P <0.01) but not in that of PMS (P >0.05). CONCLUSIONS Yijingfang is an effective drug for the treatment of asthenospermia, which can regulate the spermatogenesis, increase the percentage of PMS, and improve the total sperm motility of the patients.
Asthenozoospermia ; drug therapy ; Carnitine ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Semen ; Sperm Count ; Sperm Motility ; drug effects ; physiology ; Spermatogenesis ; drug effects

Objective: To investigate the effects of the compound preparation Jinghuosu on oligospermia and asthenospermia. METHODS: This multi-centered clinical study included 120 cases of mild to moderate idiopathic oligospermia or asthenospermia, all treated with oral Jinghuosu once a bag, bid, for 3 successive months. Before and at 1, 2 and 3 months after treatment, we detected sperm concentration, total sperm motility, progressive sperm motility and normal sperm morphology of each ejaculate, and recorded whether the patients had any adverse reactions. RESULTS: After 3 months of treatment, all the patients showed obvious improvement in semen parameters, most significantly in sperm concentration, total sperm motility, and the percentages of progressive motile sperm and morphologically normal sperm (P <0.05). No significant adverse reactions were observed during the 3 months of medication. CONCLUSIONS Jinghuosu has a significant efficacy and no obvious adverse effect in the treatment of mild to moderate oligospermia and asthenospermia.
Asthenozoospermia ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Oligospermia ; drug therapy ; Semen ; drug effects ; physiology ; Sperm Count ; Sperm Motility

Objective: To observe the effect of Huangjing Zanyu Capsule (HZC) on sperm mitochondrial membrane potential (MMP) in asthenozoospermia patients. METHODS: We assigned 70 asthenozoospermia patients to a treatment group (n = 39) and a control group (n = 31), the former treated with oral HZC at the dose of 4 capsules tid for 3 months while the latter left untreated. We obtained semen parameters from the patients and detected their sperm mitochondrial membrane potentials (MMP) by JC-1 staining and flow cytometry before and after medication, followed by comparison between the two groups. RESULTS: The total effectiveness rate was 71.05% in the treatment group and natural pregnancy was achieved in 3 cases during the medication. A total of 35 patients in the treatment group and 30 controls completed all the laboratory examinations after a 3-month observation. Compared with the controls, the patients treated with HZC exhibited significant improvement after medication in MMP (variation value: ［1.19 ± 10.36］% vs ［20.28 ± 14.21］%, P <0.01), total sperm motility (variation value: ［3.46 ± 8.67］% vs ［20.68 ± 14.12］%, P <0.01), the percentage of progressively motile sperm (variation value: ［2.26 ± 8.29］% vs ［17.58 ± 12.73］%, P <0.01), and the percentage of morphologically normal sperm (variation value: ［0.23 ± 3.48］% vs ［3.37 ± 3.99］%, P <0.01). MMP was significantly correlated with total sperm motility (r = 0.69, P <0.01), progressive sperm motility (r = 0.75, P <0.01) and normal sperm morphology (r = 0.26, P <0.01). CONCLUSIONS Huangjing Zanyu Capsule can enhance sperm mitochondrial membrane potential and sperm mitochondrial function, thus improving total sperm motility, progressive sperm motility and normal sperm morphology. It is safe and effective for the treatment of asthenospermia.
Asthenozoospermia ; drug therapy ; Capsules ; Case-Control Studies ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Flow Cytometry ; Humans ; Male ; Membrane Potential, Mitochondrial ; drug effects ; physiology ; Pregnancy ; Semen ; drug effects ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects ; ultrastructure ; Staining and Labeling

Objective: To clarify the roles of yam polysaccharide (YPS) in improving sperm viability and protecting sperm DNA integrity in vitro and provide a new approach to the treatment of oligoasthenozoospermia. METHODS: We collected samples by masturbation from 36 normal fertile males aged 27－39 years. Each sample was divided into six groups: blank control or treated with normal saline, vitamin C solution, and YPS solution at low (0.25 mg/ml), medium (1.0 mg/ml) or high concentration (5.0 mg/ml). Using eosin-Y staining, sperm hypotonic swelling (HOS) and sperm chromatin diffusion (SCD) test, we observed the effects of different concentrations of YPS on sperm viability, membrane integrity and nuclear DNA. RESULTS: After 24 and 48 hours of treatment, sperm viability was markedly reduced in the vitamin C (［28.5 ± 3.1］ and ［6.5 ± 1.2］%), low-YPS (［31.3 ± 3.5］ and ［6.5 ± 2.2］%), medium-YPS (［37.1 ± 3.5］ and ［9.5 ± 2.8］%) and high-YPS groups (［38.3 ± 3.3］ and ［9.0 ± 3.2］%) as compared with the blank control (［17.3 ± 2.1］ and ［3.2 ± 1.3］%) (P <0.01) and normal saline groups (［13.4 ± 4.1］ and ［3.1 ± 2.0］%) (P <0.01), and it was significantly higher in the medium- and high-YPS than in the vitamin C group (P <0.05 and P <0.01). The rate of sperm DNA fragmentation was remarkably decreased at 48 hours in the vitamin C (［30.5 ± 3.1］%), low-YPS (［29.4 ± 2.6］%), medium-YPS (［28.5 ± 2.3］%) and high-YPS groups (［27.9 ± 1.9］%) in comparison with the blank control (［41.7 ± 2.2］%) (P <0.01) and normal saline groups (［42.1 ± 3.3］%), markedly lower in the medium- and high-YPS than in the blank control, normal saline and vitamin C groups (P <0.05 or P <0.01), but with no statistically significant difference between the low-YPS and vitamin C groups (P >0.05). CONCLUSIONS Yam polysaccharide can improve sperm viability and protect sperm DNA integrity in vitro.
Adult ; Ascorbic Acid ; pharmacology ; DNA ; drug effects ; DNA Fragmentation ; Dioscorea ; chemistry ; Humans ; Male ; Polysaccharides ; pharmacology ; Semen Analysis ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Vitamins ; pharmacology

Objective: To investigate the effect of alpha-lipoic acid (α-LA) combined with tamoxifen citrate (TC) in the treatment of oligoasthenospermia. METHODS: From June to November 2016, we treated 60 patients with oligoasthenospermia in our Department of Andrology, 30 (the trial group) with oral α-LA (0.6 g, qd) + TC (20 mg, qd) and the other 30 (the control group) with oral L-carnitine (1g, bid) + TC (20 mg, qd). Before and after 3 months of medication, we examined the semen parameters of the patients and the levels of their seminal oxidative stress biomarkers, including methylenedioxyamphetamine (MDA) and total antioxidant capacity (TAC) in the seminal plasma. We also compared the pregnancy rate and adverse reactions between the two groups. RESULTS: Totally, 57 of the patients completed the treatment, 28 in the trial group and 29 in the control. Compared with the baseline, the patients of the trial group showed significant improvement after 3 months of medication in the semen volume (［2.50 ± 0.71］ vs ［3.37 ± 0.70］ ml, P <0.05), sperm concentration (［12.00 ± 1.65］ vs ［19.34 ± 2.04］ ×10⁶/ml, P <0.05), percentage of progressively motile sperm (PMS) (［18.01 ± 3.01］% vs ［35.41 ± 6.49］%, P<0.05), MDA level (［14.96 ± 2.76］ vs ［10.04 ± 1.04］ nmol/ml, P <0.05), and TAC in the seminal plasma (［9.83 ± 1.02］ vs ［12.25 ± 1.11］ U/ml, P <0.05), and so did the controls in the semen volume (［2.76 ± 0.67］ vs ［3.36 ± 0.93］ ml, P <0.05), sperm concentration (［11.47 ± 1.10］ vs ［17.77 ± 3.56］ ×10⁶/ml, P <0.05), percentage of PMS (［19.22 ± 1.41］ vs ［36.01 ± 5.22］ %, P <0.05), MDA level (［14.66 ± 2.75］ vs ［10.14 ± 1.01］ nmol/ml, P <0.05), and TAC in the seminal plasma (［9.84 ± 0.90］ vs ［11.14 ± 0.84］ U/ml, P <0.05). There were no statistically significant differences in the above post-medication parameters between the trial and control groups (P >0.05) except in TAC, which was markedly more improved in the former than in the latter (P <0.05), nor in the percentage of morphologically normal sperm before and after treatment in either of the two groups (P >0.05). After 3 months of treatment, 3 pregnancies were achieved in the trial group and 1 in the control (10.7% vs 3.45%, P >0.05). No obvious adverse events occurred during the treatment. CONCLUSIONS Alpha-lipoic acid combined with tamoxifen citrate can evidently improve semen parameters in oligoasthenospermia patients by relieving oxidative stress injury.
Antioxidants ; Asthenozoospermia ; drug therapy ; Biomarkers ; analysis ; Carnitine ; therapeutic use ; Drug Therapy, Combination ; Female ; Humans ; Male ; Oligospermia ; drug therapy ; Oxidative Stress ; Pregnancy ; Pregnancy Rate ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; Tamoxifen ; therapeutic use ; Thioctic Acid ; therapeutic use