Insufficiency of Mrpl40 disrupts testicular structure and semen parameters in a murine model.

Approximately 31% of patients with 22q11.2 deletion syndrome (22q11.2DS) have genitourinary system disorders and 6% of them have undescended testes. Haploinsufficiency of genes on chromosome 22q11.2 might contribute to the risk of 22q11.2DS. In this study, we used mice with single-allele deletion in mitochondrial ribosomal protein L40 (Mrpl40 +/- ) as models to investigate the function of Mrpl40 in testes and spermatozoa development. The penetrance of cryptorchidism in Mrpl40 +/- mice was found to be higher than that in wild-type (WT) counterparts. Although the weight of testes was not significantly different between the WT and Mrpl40 +/- mice, the structure of seminiferous tubules and mitochondrial morphology was altered in the Mrpl40 +/- mice. Moreover, the concentration and motility of spermatozoa were significantly decreased in the Mrpl40 +/- mice. In addition, data-independent acquisition mass spectrometry indicated that the expression of genes associated with male infertility was altered in Mrpl40 +/- testes. Our study demonstrated the important role of Mrpl40 in testicular structure and spermatozoa motility and count. These findings suggest that Mrpl40 is potentially a novel therapeutic target for cryptorchidism and decreased motility and count of spermatozoa.

[Impact of the freezing-thawing process on human sperm mitochondria].

OBJECTIVE: To study the structure and function of human sperm mitochondria before and after the freezing-thawing process. METHODS: Human sperm from healthy donors were subjected to the slow freezing-thawing process, and the sperm mitochondrion-related indexes compared before and after cryopreservation. The ultrastructural changes of the mitochondria were observed under the projection electron microscope, the mitochondrial membrane potential (MMP) and seminal adenosine triphosphate (ATP) content measured by immunofluorescence labeling and ELISA, respectively, and the sperm oxidative stress related indexes detected before and after sperm cryopreservation. RESULTS: Electron microscopy showed loose structures and widened crests of the sperm mitochondria, some with vacuole-like changes after the freezing-thawing process. The sperm after cryopreservation, compared with those before it, exhibited significantly increased contents of oxygen free radicals (ï¼»11.6 ± 3.8ï¼½% vs ï¼»9.6 ± 4.1ï¼½%, P < 0.05) and malondialdehyde (ï¼»3.2 ± 1.4ï¼½ vs ï¼»2.3 ± 1.2ï¼½ nmol/108, P < 0.05), but decreased antioxidant capacity (ï¼»0.6 ± 0.4ï¼½ vs ï¼»0.9 ± 0.4ï¼½ nmol/108, P < 0.05), superoxide dismutase activity (ï¼»0.9 ± 0.4ï¼½ vs ï¼»9.1 ± 3.9ï¼½ nmol/108, P < 0.05), MMP (ï¼»52.2 ± 6.2ï¼½% vs ï¼»55.7 ± 4.9ï¼½%, P = 0.026) and ATP production (ï¼»56.5 ± 9.0ï¼½ vs ï¼»61.3 ± 10.4ï¼½ pmol/106, P = 0.014). CONCLUSIONS: The freezing-thawing process can cause ultrastructural disorder of human sperm mitochondria, reduce their membrane potential and decrease their ATP production.

In the human sperm nucleus, nucleosomes form spatially restricted domains consistent with programmed nucleosome positioning.

In human sperm, a fraction of its chromatin retains nucleosomes that are positioned on specific sequences containing genes and regulatory units essential for embryonic development. This nucleosome positioning (NP) feature provides an inherited epigenetic mark for sperm. However, it is not known whether there is a structural constraint for these nucleosomes and, if so, how they are localized in a three-dimensional (3D) context of the sperm nucleus. In this study, we examine the 3D organization of sperm chromatin and specifically determine its 3D localization of nucleosomes using structured illumination microscopy. A fraction of the sperm chromatin form nucleosome domains (NDs), visible as microscopic puncta ranging from 40 µm to 700 µm in diameter, and these NDs are precisely localized in the post acrosome region (PAR), outside the sperm's core chromatin. Further, NDs exist mainly in sperm from fertile men in a pilot survey with a small sample size. Together, this study uncovers a new spatially-restricted sub-nuclear structure containing NDs that are consistent with NPs of the sperm, which might represent a novel mark for healthy sperm in human.

[Status quo of autologous sperm preservation in the human sperm bank in Beijing].

OBJECTIVE: To comprehensively analyze the status quo of autologous sperm preservation in the human sperm bank in Beijing and better utilize the existing resources for the preservation of male fertility. METHODS: We retrospectively analyzed the geographical data and semen quality of 251 males with autologous sperm preservation in the Human Sperm Bank, Science and Technology Research Institute, National Health and Family Planning Commission of China from July 2006 to December 2016. RESULTS: The rate of autologous sperm preservation in the Human Sperm Bank was as low as 8.76% between July 2006 and December 2010 but increased annually by 119% on average from 2011 to 2013. Of the 251 males involved, 204 (81.27%) were aged 20－39 years, 175 (69.72%) had bachelor's or master's degree, 223 (88.84%) had no child, 69 (27.49%) got less than 10 tubes of semen samples frozen, and 26 (10.36%) had their semen samples cryopreserved only once. The utilization rate of the cryopreserved sperm was only 5.58 % (n = 14). The main reason for autologous sperm preservation was carcinoma (55.78% ï¼»n = 140ï¼½), including blood cancer (22.31% ï¼»n = 66ï¼½), testicular cancer (13.15% ï¼»n = 33ï¼½) and other cancers (16.33% ï¼»n = 41ï¼½). Compared with the non-cancer males, the cancer patients had a significantly reduced mean sperm concentration (90.45 vs 60.53 ×106/ml, P < 0.05), total sperm count (311.3 vs 175.8 ×106, P < 0.05), percentage of progressively motile sperm (PMS) (49.21% vs 43.55%, P < 0.05) and recovery rate of PMS (68.13% vs 52.17%, P < 0.05). In the subgroups of testicular, blood and other cancers, the sperm concentration averaged 37.68, 57.98 and 90.69 ×106/ml, the semen volume 2.73, 2.82 and 3.41 ml, the total sperm count 93.29, 158.41 and 349.49 ×106, the percentage of PMS 45.32%, 43.47% and 44.49%, and the recovery rate of PMS 48.32%, 50.07% and 61.09%, respectively, the sperm concentration and total sperm count significantly lower in the testicular cancer patients than in the other two groups (P < 0.05). CONCLUSIONS: The number of the cases of autologous sperm preservation in Beijing is increasing year by year, and the majority of them are cancer patients. As most of the cancer patients have missed the best period for sperm preservation, sperm bank workers should endeavor to increase the public awareness of autologous sperm preservation.

[L-carnitine protects the motion parameters and mitochondrial function of human sperm in cryopreservation].

OBJECTIVE: To study the effects of L-carnitine (LC) on cryopreserved human sperm. METHODS: Ten semen samples were collected from normal sperm donors, each divided into six groups, fresh ejaculate (FE), non-LC cryopreservation (non-LC), and cryopreservation with LC at 1 mmol/L (LC-1), 2.5 mmol/L (LC-2), 5 mmol/L (LC-3) and 10 mmol/L (LC-4), respectively. The optimal concentration of LC was identified based on the motility and motion parameters of the post-thaw sperm. The plasma membrane integrity (PMI) of the sperm was assessed by eosin-nigrosin staining, their mitochondrial membrane potential (MMP) monitored by JC-1 assay, and the level of sperm ROS measured by the fluorescent probe DCFH-DA, followed by analysis of the mechanisms of LC protecting sperm against cryopreservation injury. RESULTS: Compared with the sperm in the FE group, the post-thaw sperm in the non-LC and LC groups showed significantly decreased progressive motility, average path velocity (VAP), straight line velocity (VSP) and curvilinear velocity (VCP) (P < 0.05). In comparison with the non-LC group, the LC-3 group exhibited a remarkably higher percentage of progressively motile sperm (ï¼»41.9 ± 4.6ï¼½ vs ï¼»47.0 ± 4.3ï¼½%, P = 0.0261) and VAP (ï¼»34.9 ± 2.6ï¼½ vs ï¼»38.9 ± 4.2ï¼½ µm/s, P = 0.0152), indicating that the optimal concentration of LC was 5 mmol/L. Both PMI and MMP were significantly lower in the non-LC than in the FE group (ï¼»52.7 ± 5.7ï¼½ vs ï¼»75.5 ± 5.4ï¼½%, P < 0.01 and ï¼»44.5 ± 3.5ï¼½ vs ï¼»57.3 ± 4.4ï¼½%, P < 0.01), but higher in the LC groups (ï¼»70.1 ± 8.2ï¼½% and ï¼»50.3 ± 3.4ï¼½%) than in the non-LC group (P < 0.01 and P < 0.05). The level of sperm ROS, however, was markedly higher in the non-LC than in the FE group (ï¼»12.5 ± 3.9ï¼½ vs ï¼»6.8 ± 2.4ï¼½, P < 0.01) but lower in the LC groups (ï¼»8.4 ± 5.3ï¼½%) than in the non-LC group (P = 0.05). CONCLUSIONS: L-carnitine can improve the motility and motion parameters of cryopreserved human sperm by reducing sperm ROS, enhancing sperm mitochondrial membrane potential and protecting the sperm plasma membrane.

[Fertility preservation for male adolescent cancer patients].

Fertility preservation is a hotspot of research in reproductive medicine, and that of male adolescent cancer patients is drawing even more attention from reproductive and oncologic clinicians. Both cancer and its treatment can decrease semen quality and even induce irreversible damage to fertility. Sperm cryopreservation is an effective method for fertility preservation. In the past few years, marked advances have been made in the cryopreservation, transplantation, and in vitro culture of testis tissue and stem spermatogonial cells. Although still experimental, these approaches may offer some options to those with no mature sperm in the testis. Unfortunately, very few people know and participate in the studies of fertility preservation and the utilization rate of cryopreserved sperm remains low. Therefor reproductive physicians and oncologists are required to make more efforts to search for effective fertility preservation methods for male adolescent cancer patients.

[Protective effect of Qilin Pills on the reproductive function of oligoasthenospermia rats].

OBJECTIVE: To study the protective effect of Qilin Pills (QLP) on the reproductive function of rats with oligoasthenospermia (OAS) induced by tripterygium glycosides. METHODS: Twenty-eight male SD rats were randomly divided into a normal control, an OAS model control, a low-dose QLP, and a high-dose QLP group of equal number. OAS models were made in the latter three groups by intragastrical administration of tripterygium glycosides at 40 mg per kg of the body weight per day, and meanwhile the animals in the low- and high-dose QLP groups were treated with QLP at 1.62 and 3.24 g per kg of the body weight per day, respectively, while those in the OAS model group with normal saline, all for 30 consecutive days. Then all the rats were executed for obtaining the testis weight, testis viscera index, epididymal sperm concentration and motility, reproductive hormone levels, and antioxidation indexes and observation of the histomorphological changes of the testis tissue by HE staining. RESULTS: After 30 days of intervention, the low- and high-dose QLP groups, as compared with the OAS model controls, showed significantly improved epididymal sperm concentration (ï¼»14.57 ± 3.95ï¼½ and ï¼»39.71 ± 11.31ï¼½ vs ï¼»4.71 ± 1.25ï¼½ ×106/ml, P <0.05) and motility (ï¼»3.71 ± 1.11ï¼½ and ï¼»4.29 ± 1.80ï¼½ vs ï¼»0.57 ± 0.53ï¼½%, P <0.05), increased levels of sex hormone binding globulin (SHBG) (ï¼»94.83 ± 11.17ï¼½ and ï¼»88.05 ± 9.21ï¼½ vs ï¼»56.74 ± 8.29ï¼½ nmol/L, P <0.05) and free testosterone (FT) (ï¼»27.27 ± 3.63ï¼½ and ï¼»32.80 ± 2.51ï¼½ vs ï¼»22.81 ± 2.75ï¼½ nmol/L, P <0.05), decreased level of follicle-stimulating hormone (FSH) (ï¼»1.49 ± 0.62ï¼½ and ï¼»1.12 ± 0.83ï¼½ vs ï¼»1.71 ± 0.52ï¼½ mIU/ml, P <0.05), but no significant change in the total testosterone (TT) level. Meanwhile, the level of superoxide dismutase (SOD) was markedly elevated in the low- and high-dose QLP groups in comparison with the OAS model control group (ï¼»277.14 ± 15.84ï¼½ and ï¼»299.60 ± 20.83ï¼½ vs ï¼»250.04 ± 31.06ï¼½ U/ml, P <0.05) while that of reactive oxygen species (ROS) remarkably reduced (ï¼»397.61 ± 62.71ï¼½ and ï¼»376.84 ± 67.14ï¼½ vs ï¼»552.20 ± 58.07ï¼½ IU/ml, P <0.05). HE staining showed that QLP intervention significantly increased the layers and quantity of spermatogenic cells in the testicular seminiferous tubules of the OAS rats. CONCLUSIONS: QLP can effectively protect the reproductive system of oligoasthenospermia rats by raising sperm quality, elevating reproductive hormone levels, reducing oxidative stress injury, and improving histomorphology of the testis.

[ilin Pills for oligoasthenospermia: Advances in clinical studies].

Industrialization and environmental pollution are bringing more problems to human reproduction and increasing the prevalence of male infertility. Western medicine has shown its limitations in the management of male infertility, especially that of oligoasthenospermia. Traditional Chinese medicine (TCM), however, has long and rich experiences in the treatment of oligoasthenospermia, with a large variety of medicinal prescriptions based on the TCM theories, among which Qilin Pills shows a particularly significant therapeutic effect on oligoasthenospermia, especially when combined with Western medicine. At present, published studies on Qilin Pills are mainly in the stage of clinical observation, while basic researches and studies on its relevant mechanisms are rarely seen.