Cuproptosis mediates copper-induced testicular spermatogenic cell death.

Cuproptosis, a novel mechanism of programmed cell death, has not been fully explored in the context of spermatogenic cells. This study investigated the potential involvement of cuproptosis in spermatogenic cell death using a mouse model of copper overload. Sixty male Institute of Cancer Research (ICR) mice were randomly divided into four groups that received daily oral gavage with sodium chloride (control) or copper sulfate (CuSO4) at 50 mg kg-1, 100 mg kg-1, or 200 mg kg-1, for 42 consecutive days. Mice subjected to copper overload exhibited a disruption in copper homeostasis. Additionally, significant upregulated expression of key cuproptosis factors was accompanied by a significant rise in the rates of testicular tissue cell apoptosis. Immunohistochemical analysis revealed the presence of ferredoxin 1 (Fdx1) in Sertoli cells, Leydig cells, and spermatogenic cells at various stages of testicular development, and the Fdx1-positive staining area was significantly increased in copper-overloaded mice. Mitochondrial dysfunction and decreased adenosine triphosphate levels were also observed, further implicating mitochondrial damage under cuproptosis. Further analyses revealed pathological lesions and blood-testis barrier destruction in the testicular tissue, accompanied by decreased sperm concentration and motility, in copper-overloaded mice. In summary, our results indicate that copper-overloaded mice exhibit copper homeostasis disorder in the testicular tissue and that cuproptosis participates in spermatogenic cell death. These findings provide novel insights into the pathogenic mechanisms underlying spermatogenic cell death and provide initial experimental evidence for the occurrence of cuproptosis in the testis.

Novel DNAH1 Mutation Loci Lead to Multiple Morphological Abnormalities of the Sperm Flagella and Literature Review.

The protein encoded by dynein axonemal heavy chain 1 (DNAH1) is a part of dynein, which regulates the function of cilia and sperm flagella. The mutant of DNAH1 causes the deletion of inner dynein arm 3 in the flagellum, leading to multiple morphological abnormalities of the sperm flagella (MMAF) and severe asthenozoospermia. However, instead of asthenozoospermia and MMAF, the result caused by the mutation of DNAH1 remains unknown. Here we report a male infertility patient with severe asthenozoospermia and teratozoospermia. We found two heterozygous mutations in DNAH1 (c.6912C>A and c.7076G>T) and which were reported to be associated with MMAF for the first time. We next collected and analyzed 65 cases of DNAH1 mutation and found that the proportion of short flagella is the largest, while the bent flagella account for the smallest, and the incidence of head deformity is not high in the sperm of these patients. Finally, we also analyzed 31 DNAH1 mutation patients who were treated with intracytoplasmic sperm injection (ICSI) and achieved beneficial outcomes. We hope our research will be helpful in the diagnosis and treatment of male infertility caused by DNAH1 mutation.

[Inhibitory effect of Qiangjing Tablets on the secretion of IL-1ß and TNF-ɑ from mouse Sertoli cells: A study based on the microenvironment of spermatogenesis].

OBJECTIVE: To study the effect of Qiangjing Tablets (QJT) on the secretion of the inflammatory cytokines IL-1ß and TNF-ɑ from Sertoli cells in infertile mice based on the microenvironment of spermatogenesis. METHODS: We isolated and cultured mouse Sertoli cells, established the model of Ureaplasma urealyticum (UU) infection in the cells, and treated the cells with QJT at the concentrations of 2.5%, 5% and 10% in the serum. After modeling, we determined the contents of IL-1ß and TNF-ɑ in the supernatant of the cells by ELISA and examined the effect of QJT on the secretion of the inflammatory factors from the Sertoli cells by analyzing the dose-effect and time-effect relationships of the drug. RESULTS: In comparison with the blank control, the UU-infected Sertoli cells showed significantly increased secretion of IL-1ß and TNF-ɑ (P < 0.05), the former reaching the peak value in 12 hours and the latter in 24 hours, followed by a downward trend. The secretion of IL-1ß was remarkably inhibited in the 5% and 10% QJT groups (P < 0.05) and that of TNF- ɑ in the 10% QJT group compared with those in the UU infection model group (P < 0.05). CONCLUSIONS: The secretion of IL-1ß and TNF-ɑ is significantly increased in the UU-infected Sertoli cells, and that of IL-1ß negatively correlated with time. QJT-containing serum can inhibit the secretion of IL-1ß and TNF-ɑ from Sertoli cells, and the inhibitory effect of IL-1 ß is most significant at 5% and 10% and that of TNF- ɑ at 10%.

[Effect of Qiangjing Tablets on the MAPK signaling pathway in SD rats with asthenospermia].

OBJECTIVE: To investigate the effects of Qiangjing Tablets (QJT) on sperm quality and the MAPK signaling pathway in the SD rat model of asthenospermia (AS). METHODS: A 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-ß1 in the semen measured by ELISA. RESULTS: The 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 (ï¼»9.20 ± 3.07ï¼½ vs ï¼»42.20 ± 9.17ï¼½ %, P<0.01), but decreased level of TGF-ß1 in the semen (ï¼»627.67 ± 26.07ï¼½ vs ï¼»566.73 ± 68.44ï¼½ 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 (ï¼»42.20 ± 9.17ï¼½ vs ï¼»21.60 ± 5.94ï¼½ %, P<0.01; ï¼»42.20 ± 9.17ï¼½ vs ï¼»33.95 ± 6.39ï¼½ %, P<0.05). The concentration of TGF-ß1 in the semen was markedly lower in the high-dose QJT than in the AS model control group (ï¼»621.78 ± 30.80ï¼½ vs ï¼»566.73 ± 68.44ï¼½ ng/ml, P < 0.05). CONCLUSIONS: Qiangjing Tablets could improve semen quality in asthenospermia rats by acting against oxidative stress.

[Qilan Capsules plus androgen-deprivation therapy for Qi-deficiency blood-stasis type of prostate cancer after castration].

OBJECTIVE: To observe the synergistic effect of Qilan Capsules in the treatment of the patient with Qi-deficiency blood-stasis type of prostate cancer receiving androgen-deprivation therapy after castration. METHODS: This randomized controlled double-blind study included 246 cases of Qi-deficiency blood-stasis type of prostate cancer after castration, which were randomly divided into an experiment and a control group of equal number to be treated with Qilan Capsules + androgen-deprivation and placebo + androgen-deprivation, respectively. After 6 months of treatment, we compared the International Prostate Symptoms Scores (IPSS), TCM Symptoms Scores (TCMSS), maximal urine flow rate (Qmax), and the level of serum prostate-specific antigen (PSA) between the two groups of patients. RESULTS: Statistically significant differences were observed between the experiment and control groups in the syndrome classification-based efficacy (87.7% vs 67.9%, P <0.05) and total effectiveness rate (86.0% vs 71.6%, P <0.05). Compared with the baseline, the experiment group showed remarkable improvement after treatment in TCMSS (17.1±5.1 vs 8.3±4.0, P <0.05), IPSS (17.7±7.5 vs 11.4±4.6, P <0.05), and Qmax (ï¼»10.9±4.3ï¼½ ml/s vs ï¼»14.7±3.7ï¼½ ml/s, P <0.05), and so did the control group (16.8±5.2 vs 11.5±5.2, 17.8±6.7 vs 14.6±5.8, and ï¼»11.0±4.3ï¼½ ml/s vs ï¼»12.0±4.1ï¼½ ml/s, P <0.05). The above three parameters were even more markedly improved in the former than in the latter group (P <0.05). However, there was no statistically significant difference between the two groups in the improvement of the PSA level after treatment (P >0.05). CONCLUSIONS: Qilan Capsules can significantly enhance the effect of androgen-deprivation therapy in the treatment of Qi-deficiency blood-stasis type of prostate cancer after castration though cannot obviously improve the PSA level.

[Umbilicus application of Huobi Powder for ED with kidney deficiency dampness: A randomized controlled clinical trial].

OBJECTIVE: To prove the clinical efficacy of umbilicus application of Huobi Powder for the treatment of erectile dysfunction (ED) with the traditional Chinese medicine (TCM) syndrome of kidney deficiency dampness by related clinical indexes. METHODS: This randomized controlled double-blind clinical study included 72 ED patients with the TCM syndrome of kidney deficiency dampness treated by 12-hour application of Huobi Powder (the trial group, n = 36) or placebo (the control group, n = 36) to the umbilicus qd for 28 consecutive days. We recorded the IIEF-5 and Erection Quality Scale (EQS) scores, TCM syndrome indexes, radial rigidity of the erectile penis, and the angle of penile erection before and after treatment. We established a database with the collected data and performed statistical analysis with the SPSS21.0 software. RESULTS: Statistically significant differences were observed after treatment between the trial and control groups in the TCM syndrome-based efficacy (69.44% vs 36.11%, P <0.01) and Western medicine-based clinical effectiveness (77.78% vs 36.11%, P <0.01). The trial group, as compared with the control, showed remarkably decreased TCM syndrome indexes (18.19 ± 9.12 vs 12.97 ± 11.92, P <0.05), increased IIEF-5 score (13.83 ± 4.91 vs 18.67 ± 3.09, P <0.01), radial rigidity of the erectile penis (ï¼»53.43 ± 11.05ï¼½% vs ï¼»71.96 ± 12.92ï¼½%, P <0.01) and the angle of penile erection (ï¼»42.15 ± 11.77ï¼½% vs ï¼»66.96 ± 12.34ï¼½%, P <0.01), but no significant difference in the EQS score (37.11 ± 16.70 vs 35.33 ± 14.11, P >0.05). CONCLUSIONS: Umbilicus application of Huobi Powder has a definite clinical effect on ED with the TCM syndrome of kidney deficiency dampness.

[Inhibitory effect of Qiangjing Tablets on the Fas/FasL pathway of cell apoptosis in male SD rats with infertility].

OBJECTIVE: To observe the effects of Qiangjing Tablets (QJT) on the semen quality and Fas/FasL signaling pathway in male SD rats with infertility. METHODS: Models of infertility were made in 50 male SD rats by intragastric administration of Tripterygium (GTW) for 3 weeks, and another 20 rats were taken as blank controls. Then 40 successfully established rat models were randomly divided into four groups, model control, low-dose QJT, medium-dose QJT, and high-dose QJT, the latter three groups treated intragastrically with QJT at 58 mg, 105 mg, and 233 mg per kg of the body weight per day, respectively. After 4 weeks of medication, the rats were killed for examination of semen quality and determination of the expression of the apoptosis factor FasL in the testis tissue. RESULTS: Compared with the blank controls, the model rats showed significant decreases in sperm concentration ([71.99 ± 9.72] vs [10.94 ± 3.58] x 106/ml, P < 0.01), motility ([48.95 ± 4.10] vs [9.31 ± 5.79]%, P < 0.01), and viability ( [82.06 ± 6.16] vs [24.03 ± 6.93]%, P < 0.01). In comparison with the model controls, the rats in the QJT groups exhibited remarkably increased sperm concentration, motility, and viability, more significantly in the high-dose group ([59.66 ± 4.53] x 106/ml, [35.45 ± 5.21] %, and [61.97 ± 9.75]%) and medium-dose group ([40.89 ± 4.90] x 106/ml, [24.41 ± 4.79]%, and [60.06 ± 10.62]%) (P < 0.05 or P < 0.01). The expression of FasL was markedly reduced in the low-, medium-, and high-dose QJT groups (0.5215 ± 0.0189, 0.5371 ± 0.0364, and 0.4556 ± 0.0215) as compared with that of the model controls (0.5989 ± 0.0448 ) (P < 0.05 or P < 0.01). CONCLUSION: By upregulating the Fas/FasL signaling pathway, Tripterygium glycosides may induce the apoptosis of spermatogenic cells and reduce sperm concentration, motility and viability, resulting in infertility. The Chinese medicine Qiangjing Tablets can improve the reproductive function of male rats by decreasing the expression of the apoptosis factor FasL in the testis.

[Longbishu Capsule combined with mesylate doxazosin: an efficacious therapy for benign prostatic hyperplasia].

OBJECTIVE: To assess the clinical effect and safety of the Chinese medicine Longbishu Capsule combined with mesylate doxazosin in the treatment of benign prostatic hyperplasia (BPH) of the kidney deficiency and blood stagnation type. METHODS: This was a randomized, double-blind, double-simulation control study. We equally assigned 60 men diagnosed with BPH of the kidney deficiency and blood stagnation type to an experimental and a control group, the former treated with mesylate doxazosin plus Longbishu Capsule and the latter with mesylate doxazosin plus placebo. We compared the International Prostate Symptom Score (IPSS), quality of life (QOL), Chinese symptom score (CSS), maximal urinary flow rate (Qmax), and prostate volume between the two groups of patients before and after 6 months of medication. RESULTS: After treatment, there were 5 cured cases, 13 markedly effective cases, 9 effective cases, 1 ineffective case, and 2 eliminated cases in the experimental group, as compared with 2 cured cases, 8 markedly effective cases, 10 effective cases, 7 ineffective cases, and 3 eliminated cases in the control group. The total effectiveness rate was obviously higher in the former (96.4%) than in the latter (74.1%). IPSS, Qmax, and CSS were improved in both of the groups after medication, even more significantly in the experimental than in the control group (IPSS: 15.22 ± 2.98 vs 18.15 ± 5.88, P <0.05; Qmax: [13.56 ± 2.26] ml/s vs [11.78 ± 2.97] ml/s, P <0.05; CSS: 6.18 ± 2.13 vs 9.52 ± 3.15, P <0.05). Because of the difference in the QOL score between the two groups at the baseline (P = 0.038 <0.05), no more comparison was made in this aspect after treatment. CONCLUSION: The combination of Longbishu Capsule with mesylate doxazosin is safe and effective for the treatment of BPH.

Intracytoplasmic sperm injection (ICSI) outcomes of azoospermia with different causes: 107 cases report.

This retrospective analysis compared the outcomes of fertilization and pregnancy rates of 107 azoospermia patients treating with intracytoplasmic sperm injection (ICSI). Sperms were obtained by testicular biopsy surgery, with which we used in ICSI subsequently. The outcomes were compared by different kinds of causes leading to azoospermia in the 107 cases. 69 cases of obstructive azoospermia and 38 cases non-obstructive, the fertilization rates were 61.94% and 53.47% respectively, and pregnancy rates were 67.65% and 52.63%. 78 cases with normal volume testes and 29 cases with small testes, the fertilization rates were 70.93% and 48.80% respectively, and pregnancy rates were 66.25% and 50.00%. There was significant difference in fertilization rates between obstructive, non-obstructive and normal volume testes, small testes (P < 0.05), but no significant difference in pregnancy rates (P > 0.05). The pregnancy rate was significant difference between female age < 32 and ≥ 32 whatever the cause of azoospermia was (P < 0.05). Our study reveals that obstructive azoospermia and normal volume testes have higher fertilization rates in ICSI, but the pregnancy rates are only related to female age.

[Yuleshu oral mixture combined with conventional therapy for chronic prostatitis].

OBJECTIVE: To evaluate the clinical effects of Yuleshu oral mixture combined with conventional therapy on chronic prostatitis. METHODS: Eighty-eight patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) were equally randomized to a control and an experimental group to receive conventional therapy (oral antibiotics, alpha blockers, proprietary Chinese medicine for activating blood circulation and massage of the prostate) and conventional therapy combined with Yuleshu oral mixture respectively. Before and after treatment, the severity of symptoms and sexual function of the patients were evaluated using NIH-CPSI and IIEF-5, their anxiety, depression and other emotional problems assessed with Hamilton Depression Rating Scale (HAMD) and Hamilton Anxiety Scale (HAMA), and the results subjected to statistical analysis. RESULTS: Both the experimental and control groups showed significant improvement in prostatitis symptoms and sexual function after treatment as compared with the baseline (P < 0.01), even more significant in the former than in the latter group, especially in pain symptoms (7.89 +/- 2.82 vs 10.41 +/- 2.55, P < 0.01). Before and after treatment, the HAMA and HAMD score had no significant difference in the control, but there was significant difference in the experimental group. The experimental group exhibited remarkably higher scores after than before treatment on HAMA (24.30 +/- 5.07 vs 13.80 +/- 3.62, P < 0.01) and HAMD (23.81 +/- 5.01 vs 16.23 +/- 5.93, P < 0.01), but not the control group (P > 0.05). CONCLUSION: Yuleshu oral mixture can effectively relieve anxiety, depression and other psychological problems in CP/CPPS patients, and improve their clinical symptoms as well. Therefore, it is an effective drug for chronic prostatitis.

[Correlation of IPSS with IL-8 and COX-2 levels in patients with benign prostatic hyperplasia and prostatitis].

OBJECTIVE: To investigate the correlation of International Prostate Symptom Score (IPSS) with the levels of interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2) in the prostate tissue and expressed prostatic secretion (EPS) in patients with benign prostatic hyperplasia (BPH) complicated by prostatitis. METHODS: We divided 80 BPH patients to be treated by transurethral resection of the prostate (TURP) into a simple BPH group (n = 30) and a BPH with prostatitis group (n = 50) based on the pathologic features. We statistically analyzed IPSS and the levels of IL-8 and COX-2 in EPS before surgery and the IL-8 and COX-2 levels in the prostate tissue after surgery. RESULTS: IPSS was positively correlated with the IL-8 and COX-2 levels in the prostate tissue and EPS of the BPH patients, moderately in the simple hyperplasia group (r > 0.5) and highly in the other (r > 0.8). The levels of IL-8 and COX-2 in the prostate tissue and EPS were significantly higher in the BPH with prostatitis group than in the simple BPH group (P < 0.05). CONCLUSION: The levels of IL-8 and COX-2 in EPS indirectly reflect those in the prostate tissue. IPSS and the levels of IL-8 and COX-2 in EPS can help determine whether BPH is complicated by histological prostatic inflammation.

[Association between leptin gene promoter methylation and type 2 diabetes mellitus].

OBJECTIVE: To assess the association between leptin gene promoter methylation and serum leptin concentrations in patients with impaired glucose regulation (IGR) and type 2 diabetes mellitus (T2DM). METHODS: Methylation status of leptin gene promoter was determined with methylation-specific polymerase chain reaction. Serum leptin concentrations were determined using enzyme-linked immunosorbent assay. RESULTS: Among three groups of individuals with different levels of glucose, the methylation rates of leptin gene in IGR and T2DM groups were 43.6 % and 31.5 %, respectively, which were significantly lower than that of healthy subjects (59.2%; Chi-square=22.499 and 5.109, respectively, P<0.05). A lower methylation rate was also observed in T2DM group compared with IGR group (Chi-square=3.962, P<0.05). Leptin levels in both T2DM and IGR groups were elevated compared with normoglycemic subjects, but only T2DM group was significantly higher (q=6.81, P<0.01). Linear regression analysis indicated that serum leptin concentrations has increased along with declining of DNA methylation rate (r=-0.95, P<0.01). CONCLUSION: Lower levels of leptin gene promoter DNA methylation and serum leptin concentrations are associated with the development of diabetes. Measurement of the methylation status of leptin gene promoter and expression can facilitate early intervention of the disease.

[Clinical features of benign prostatic hyperplasia complicated by chronic prostatitis].

OBJECTIVE: To explore the clinical characteristics of benign prostatic hyperplasia (BPH) complicated by chronic prostatitis (CP). METHODS: A total of 120 cases of BPH pathologically confirmed after transurethral resection of the prostate (TURP) were assigned to a BPH group (n=75) and a BPH + CP group (n=45) according to whether they were complicated by CP. The total prostatic volume (TPV) and PSA density (PSAD) were calculated and statistically analyzed based on the results of transrectal ultrasonography and f-PSA, t-PSA and f-PSA/t-PSA tests before surgery. RESULTS: The BPH group showed a significantly upward tendency in f-PSA and t-PSA (P < 0.05) with the increase of age or prostate volume, but not significantly in PSAD and f-PSA/t-PSA (P > 0.05). In comparison, the BPH + CP group exhibited remarkable increases in f-PSA, t-PSA and PSAD (P < 0.05) but not in fPSA/t-PSA (P > 0.05). ROC curve analyses of various indexes showed the area under the curve to be 0.644, 0.628 and 0.624 for f-PSA, t-PSA and PSAD, respectively, all between 0.5 and 0.7. CONCLUSION: BPH is frequently associated with CP. Clinically, high f-PSA, t-PSA and PSAD are important but not sure indicators of BPH complicated by CP.