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Background Human sperm cryopreservation is a simple and effective approach for male fertility preservation. Methods To identify potential proteomic changes in this process, data-independent acquisition (DIA), a technology with high quantitative accuracy and highly reproducible proteomics, was used to quantitatively characterize the proteomics of human sperm cryopreservation. Results A total of 174 significantly differential proteins were identified between fresh and cryoperservated sperm: 98 proteins decreased and 76 proteins increased in the cryopreservation group. Bioinformatic analysis revealed that metabolic pathways play an important role in cryopreservation, including: propanoate metabolism, glyoxylate and dicarboxylate metabolism, glycolysis/gluconeogenesis, and pyruvate metabolism. Four different proteins involved in glycolysis were identified by Western blotting: GPI, LDHB, ADH5, and PGAM1. Conclusions Our work will provide valuable information for future investigations and pathological studies involving sperm cryopreservation.

To study the structure and function of human sperm mitochondria before and after the freezing-thawing process.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.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 (［11.6 ± 3.8］% vs ［9.6 ± 4.1］%, P0.05) and malondialdehyde (［3.2 ± 1.4］ vs ［2.3 ± 1.2］ nmol/108, P0.05), but decreased antioxidant capacity (［0.6 ± 0.4］ vs ［0.9 ± 0.4］ nmol/108, P0.05), superoxide dismutase activity (［0.9 ± 0.4］ vs ［9.1 ± 3.9］ nmol/108, P0.05), MMP (［52.2 ± 6.2］% vs ［55.7 ± 4.9］%, P = 0.026) and ATP production (［56.5 ± 9.0］ vs ［61.3 ± 10.4］ pmol/106, P = 0.014).The freezing-thawing process can cause ultrastructural disorder of human sperm mitochondria, reduce their membrane potential and decrease their ATP production.

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.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.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% ［n = 140］), including blood cancer (22.31% ［n = 66］), testicular cancer (13.15% ［n = 33］) and other cancers (16.33% ［n = 41］). Compared with the non-cancer males, the cancer patients had a significantly reduced mean sperm concentration (90.45 vs 60.53 ×10⁶/ml, P0.05), total sperm count (311.3 vs 175.8 ×106, P0.05), percentage of progressively motile sperm (PMS) (49.21% vs 43.55%, P0.05) and recovery rate of PMS (68.13% vs 52.17%, P0.05). 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 (P0.05).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.

To study the effects of L-carnitine (LC) on cryopreserved human sperm.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.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) (P0.05). In comparison with the non-LC group, the LC-3 group exhibited a remarkably higher percentage of progressively motile sperm (［41.9 ± 4.6］ vs ［47.0 ± 4.3］%, P = 0.0261) and VAP (［34.9 ± 2.6］ vs ［38.9 ± 4.2］ 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 (［52.7 ± 5.7］ vs ［75.5 ± 5.4］%, P0.01 and ［44.5 ± 3.5］ vs ［57.3 ± 4.4］%, P0.01), but higher in the LC groups (［70.1 ± 8.2］% and ［50.3 ± 3.4］%) than in the non-LC group (P0.01 and P0.05). The level of sperm ROS, however, was markedly higher in the non-LC than in the FE group (［12.5 ± 3.9］ vs ［6.8 ± 2.4］, P0.01) but lower in the LC groups (［8.4 ± 5.3］%) than in the non-LC group (P = 0.05).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.

Testosterone supplementation may be effective for the treatment of hypogonadism in men with type 2 diabetes mellitus (T2DM), but the evidence from randomized controlled trials (RCTs) is inconclusive. We aimed to systematically summarize results from intervention studies and assess the effects of testosterone supplementation therapy (TST) on lipid metabolism in RCTs of hypogonadal men with T2DM by meta-analysis. PubMed, Embase, and Cochrane Library databases were searched for studies reporting the effect of TST on lipid metabolism in hypogonadal men with T2DM until December 31, 2016. Seven RCTs from 252 trials, enrolling a total of 612 patients in the experimental and control groups with a mean age of 58.5 years, were included in this study. The pooled results of the meta-analysis demonstrated that TST significantly decreased TC and TG levels in hypogonadal men with T2DM compared with the control group, with mean differences (MDs) of -6.44 (95% CI: -11.82 to -1.06; I2 = 28%; p = 0.02) and -27.94 (95% CI: -52.33 to -3.54; I2 = 76%; p = 0.02). Subgroup analyses revealed that the heterogeneity (I2 = 76%) of TG originated from different economic regions, in which economic development, genetic and environmental factors, and dietary habits affect lipid metabolism of human, with a decrease (I2 = 45%) in developed countries. Additionally, subgroup analyses showed that TST increased HDL-C level in developing countries compared with the control group (MD = 2.79; 95% CI: 0.73 to 4.86; I2 = 0%; p = 0.008), but there was no improvement in developed countries (MD = 1.02; 95% CI: -4.55 to 6.60; I2 = 91%; p = 0.72). However, LDL-C levels were not improved consistently. Because the relationship between lipid metabolism and atherosclerosis is unequivocal, TST, which ameliorates lipid metabolism, may decrease the morbidity and mortality of cardiovascular disease in hypogonadal men with T2DM by preventing atherogenesis.

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., Summary: Programmed gene expression is the key to life, confirmed experimentally in this study, which shows that nucleosomes in human sperm are specifically localized in genomic regions, guiding programmed gene expression.

BACKGROUND Fertility preservation is very important for male cancer patients, especially adolescents. Unfortunately, the use of fertility preservation is very low among Chinese male cancer patients. Additionally, the cumulative rate of frozen sperm use is also low. MATERIAL AND METHODS We performed a retrospective study by collecting available information at the Human Sperm Bank, National Research Institute for Family Planning from July 2006 to December 2017 to examine the data in China. RESULTS A total 145 male cancer patients underwent sperm cryopreservation. The patients were 29.3±6.9 years old, and 6.2% (9 out of 145) of the patients were adolescents under the age of 18 years old. As of June 2018, only 9.7% (14 out of 145) of patients returned to use their cryopreserved sperm for assisted reproduction technology (ART). Of the 33 ART cycles, conceptions were achieved in 51.5% (17 out of 33), and the rate of patients who had a baby was 71.4% (10 out of 14). The data indicate men with testicular cancer or leukemia had lower total sperm counts and recovery rate of progressive sperm than did men with other types of cancer, while men with sarcoma had the lowest progressive sperm. CONCLUSIONS The physician should make an effort to promote fertility preservation for male cancer patients in China. And patients with testicular cancer and leukemia require additional attention.

To study the protective effect of Qilin Pills (QLP) on the reproductive function of rats with oligoasthenospermia (OAS) induced by tripterygium glycosides.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.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 (［14.57 ± 3.95］ and ［39.71 ± 11.31］ vs ［4.71 ± 1.25］ ×10⁶/ml, P0.05) and motility (［3.71 ± 1.11］ and ［4.29 ± 1.80］ vs ［0.57 ± 0.53］%, P0.05), increased levels of sex hormone binding globulin (SHBG) (［94.83 ± 11.17］ and ［88.05 ± 9.21］ vs ［56.74 ± 8.29］ nmol/L, P0.05) and free testosterone (FT) (［27.27 ± 3.63］ and ［32.80 ± 2.51］ vs ［22.81 ± 2.75］ nmol/L, P0.05), decreased level of follicle-stimulating hormone (FSH) (［1.49 ± 0.62］ and ［1.12 ± 0.83］ vs ［1.71 ± 0.52］ mIU/ml, P0.05), 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 (［277.14 ± 15.84］ and ［299.60 ± 20.83］ vs ［250.04 ± 31.06］ U/ml, P0.05) while that of reactive oxygen species (ROS) remarkably reduced (［397.61 ± 62.71］ and ［376.84 ± 67.14］ vs ［552.20 ± 58.07］ IU/ml, P0.05). HE staining showed that QLP intervention significantly increased the layers and quantity of spermatogenic cells in the testicular seminiferous tubules of the OAS rats.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.目的： 研究麒麟丸对雷公藤多苷所致少弱精子症模型雄性大鼠的生殖功能保护作用。方法： 取28只雄性SD大鼠，随机分成正常对照组、模型组、麒麟丸低、高剂量组，除正常对照组外，其他各组灌服雷公藤多苷40 mg/(kg·d)，同时麒麟丸给药组同日给予不同剂量麒麟丸保护，低剂量组给予麒麟丸1.62 g/(kg·d)，高剂量组给予麒麟丸3.24 g/(kg·d)。连续给药30 d后，处死大鼠，称重大鼠睾丸，计算睾丸脏器指数，检测附睾精子浓度、活动率，检测生殖激素水平，评估抗氧化指标，HE切片观察大鼠睾丸组织形态学改变。结果： 实验结束后，麒麟丸低、高剂量组精子浓度分别为(14.57±3.95) ×10⁶/ml 、(39.71±11.31) ×10⁶/ml ，精子活力为(3.71±1.11)%、 (4.29±1.80)% 皆较模型组(4.71±1.25)×10⁶/ml、(0.57±0.53)%有明显提高（P0.05）；与模型组FSH (1.71±0.52) mIU/ml比较，麒麟丸低、高剂量组FSH下降，分别为(1.49±0.62) mIU/ml 、(1.12±0.83) mIU/ml ；fT水平较模型组升高(27.27±3.63) nmol/L 、(32.80±2.51) nmol/L vs (22.81±2.75) nmol/L，SHBG水平升高(94.83±11.17) nmol/L 、(88.05±9.21) nmol/L vs (56.74±8.29) nmol/L，差异皆有显著性（P0.05），但T水平无明显变化（P0.05）。与模型组比较，麒麟丸低、高剂量组SOD水平升高(277.14±15.84) U/ml 、(299.60±20.83) U/ml vs (250.04±31.06) U/ml；ROS水平下降(397.61±62.71) IU/ml 、(376.84±67.14) IU/ml vs (552.20±58.07) IU/ml（P0.05）。与模型组比较，低、高剂量组HE染色显示麒麟丸能增加少弱精子症大鼠睾丸生精小管内各级生精细胞层次和数量。结论： 麒麟丸能明显提高少弱精子症大鼠精子质量，改善生殖激素水平，减轻氧化应激损伤，改善大鼠睾丸组织形态，对生殖系统有较好的保护作用。.

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.随着人类工业化进程快速发展以及环境污染等情况的出现，人类生殖面临诸多困境，男性不育症患病率逐年升高，西医在治疗男性不育症，尤其是少弱精子症方面有诸多局限。祖国传统中医药在治疗少弱精子症方面历史悠久，经验丰富；形成了以中医理论为基础的大批药剂，在治疗少弱精子症方面发挥着重要作用。麒麟丸作为其中的典型代表，在治疗少弱精子症方面有显著疗效，尤其和西药联合应用疗效更加明显。目前，麒麟丸研究主要集中于临床疗效观察阶段，基础研究较少，缺乏相关机制的研究。.

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.生育力保护是目前生殖医学领域的研究热点。男性青少年肿瘤患者的生育力保护问题更应该引起肿瘤科及生殖科医师的高度关注。肿瘤及其治疗过程都会降低精液质量，甚至引起生育力不可逆的损伤。目前自体精液保存仍是生育力保护最为成熟的方式。睾丸组织和精原干细胞冻存、移植及体外培养近年来虽取得了很大发展,这种保护方式为睾丸内无成熟精子的人群提供了选择可能，但其进入临床应用阶段尚需更多的研究证据。目前，国内外对生育力保护的研究现状表现为知情人少、参与人数少、冻存精子最终使用率低等现象，该现状的改善就需要加强男性青少年肿瘤患者生育力保护的全面而有效的探索。.

At present, the treatment of oligoasthenospermia with western medicine is ineffective. Qilin pill (QLP) is a Chinese traditional medicine for treating male infertility. Recent multicenter clinical studies in China reported that QLPs markedly improved sperm quality. However, the mechanism of action of QLPs on oligoasthenospermia remains unknown. In this study, we investigated the mechanistic basis for improvement of semen parameters and reversal of testis damage by QLPs in a rat model of oligoasthenospermia induced by treatment with tripterygium glycosides (TGs) (40 mg/kg) once daily for 4 weeks. Rats were administered QLPs (1.62 g/kg or 3.24 g/kg) each day for 60 days, with untreated animals serving as controls. The concentration and motility of sperm extracted from rat epididymis were determined, whereas histopathological examination and immunohistochemical apoptosis analysis of rat testes was performed. Expression profiles of apoptosis-related genes were determined by microarray analysis; the results were validated by quantitative real-time PCR, western blotting, and immunohistochemistry. Sperm concentration and motility in the QLP treatment group were increased relative to those in control rats. Testis tissue and DNA damage were reversed by QLP treatment. The improvement function of QLPs on sperm and testis works mainly by suppressing mitochondrial apoptosis in the testis via modulation of B cell lymphoma (Bcl)-2, Bcl-2-associated X protein (Bax), cytochrome C, caspase-9 and caspase-3 expression. QLPs could improve sperm quality and testis damage in a rat model of oligoasthenospermia by inhibiting the Bax-Caspase-9 apoptosis pathway and exerting therapeutic effects.

To investigate the dynamic transfer characteristics of low geosynthetic-reinforced embankments supported by CFG piles under high-speed train load, a numerical study has been conducted through dynamic finite element method on basis of the dynamic field test on a cross-section of Beijing-Shanghai high-speed railway. The comparative analysis on results of numerical study and field test indicated the distribution characteristics of vertical dynamic stress induced by high-speed train load in subgrade soil under railway line. The numerical results also suggested a high stress area in subgrade where vertical dynamic stress is over 1kPa. Conclusions of this work can provide reference for both design and estimation of long-term settlement of low geosynthetic-reinforced embankments supported by CFG piles for high-speed railway.

17 half-sib families and 51 full-sib families of Hyriopsis cumingii were constructed by nested design.40 individuals were randomly selected from each of these full-sib families when the individuals in the cages reached 1 cm,and the number was 2 040 in all.Shell length,shell height,shell width and body weight were measured of these individuals,and these data were used for genetic analysis.The heritability of these four traits was 0.356±0.047,0.488±0.060,0.453±0.055 and 0.518±0.050 respectively.The phenotype correlation and genetic correlation was between 0.476-0.709 and 0.574-0.868.There was enough genetic variance for selective breeding and when we have selective breeding program on weight,the other three traits would have some genetic improvement at the same time.