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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 investigate the prevalence of metabolic syndrome (MS) and its relationship with reproductive hormones in adult males in China.Using the cluster and stratified sampling methods, we conducted an investigation among 3 600 adult males aged over 20 years in Hebei, Shaanxi and Jiangsu provinces. We obtained their reproductive hormone levels, biochemical indicators and basic body indexes and compared them between the two groups of subjects.A total of 3 332 valid serum samples were collected, which revealed a prevalence rate of MS of 38.5% in the adult males. Compared with the non-MS subjects, the MS males showed a significantly higher free testosterone index (FTI, 0.39 ± 0.15 vs 0.45 ± 0.19, P0.01) but lower levels of total testosterone (TT, ［16.35 ± 4.78］ vs ［13.37 ± 4.23］ nmo/L, P0.01) and sex hormone-binding globulin (SHBG, ［47.13 ± 20.50］ vs ［33.32 ± 14.91］ nmol/L, P0.01) and testosterone secretion index (TSI, 3.64 ± 1.92 vs 3.14 ± 1.80, P0.01). There were no statistically significant differences between the two groups in the levels of calculated free testosterone (cFT) and LH (P0.05). Spearman rank correlation analysis showed that the levels of serum TT and SHBG were correlated with all the indicators of MS (P0.01) except systolic blood pressure (SBP), while that of cFT only with the levels of high-density lipoprotein (HDL), fasting blood glucose (FBG) and SBP (P0.01). After adjusted for age, smoking, drinking, and body mass index (BMI), all the MS indicators were significantly associated with the SHBG level (P0.01), but not high blood pressure with the serum TT level (P0.05). After adjusted for the age, smoking, drinking, BMI and TT, the serum SHBG level was the main independent risk factor for MS (OR: 0.965, 95% CI: 0.958－0.973, P0.01).The level of serum cFT is not correlated with while that of SHBG is the main independent risk factor for metabolic syndrome in adult males in China.

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。.

To investigate the relationship among serum reproductive hormone levels, serum homocysteine (Hcy) levels, metabolic syndrome (MS), and the components of MS in middle-aged and elderly males.Using the cluster and stratified sampling methods and a unified structured questionnaire, we conducted a survey among 948 men aged 40 - 80 years in the rural community, measured their basic physical parameters, and obtained their reproductive hormone levels, serum Hcy concentrations, and metabolism-related indicators. We collected 868 valid questionnaires along with their serum samples, divided the subjects into an MS and a non-MS control group in a 1:1 ratio, and measured their serum Hcy concentrations.Among the subjects included, 132 were diagnosed with MS. Nonparametric tests showed statistically significant differences between the MS and non-MS groups in the waist circumference (WC), waist-hip ratio (WHR), body mass index (BMI), systolic blood pressure (SBP), and diastolic blood pressure (DBP) (P0.05), but not in age (P0.05). Significant differences were also observed between the two groups in the levels of serum tT, SHBG, LH, and FTI (P0.05) , but not in the concentrations of serum Hcy (P0.05). The concentration of serum Hcy exhibited no correlation with BMI, SBP, DBP, FBG, TG, and HDL-C (P0.05) and had no influence on MS.The concentration of serum Hcy is not significantly correlated with MS, nor with its components. The levels of male serum reproductive hormones are associated both with MS and with its components.

To establish the methods for training green-hand laboratorians in standard semen analysis and evaluating the training results, and afford some reference for internal quality control and training in andrology laboratories in China.We trained the green-hand technicians in standard semen analysis recommended by WHO Laboratory Manual for the Examination and Processing of Human Semen (5th ed), and evaluated the training results by assessing the trainees' performance in the examination of sperm concentration and motility by statistic analysis.After training, the coefficients of variation of the semen samples with high, middle and low sperm concentration achieved by the trainees were 7.72% and 3.38% and 4.49%, and those with high, middle and high motility were 7.82%, 8.09% and 6.62%, respectively. We used Bland-Altman's method to evaluate the consistency between the results obtained by the trainees and those by the trainers. For sperm concentration, 4.77% of the datum points were out of the 95% consistency interval, and the absolute value of the biggest difference between the trainees and trainers was 8 x 10(6)/ml within the 95% consistency interval. For sperm motility, 7.15% of the data points were out of the 95% consistency interval, and the absolute value of the biggest difference between the trainees and trainers was 10% within the 95% consistency interval. Two-way analysis of variance showed no significant differences in the results of sperm concentration and motility analyses between the trainees and trainers (P0.05).Training in standard semen analysis significantly improved the precision of semen analysis among the green-hand laboratorians. The training and assessment methods we established proved to be effective and feasible.

To investigate the mechanism of serum testosterone reduction, its relationship with metabolism, changes in the number and morphology of Leydig cells and endocrine function in aging male rats.The levels of serum total testosterone (tT), LH, FSH, HDL, LDL, TG, TC, Glu, INS, IRG and LP were determined in young (9 mo) and aging rats (12, 15, 18 and 21 mo), with 6 in each group. The morphological changes of Leydig cells were observed under the microscope. The concentrations of testosterone secreted from the cultured Leydig cells with the stimulation of hCG and Forskolin were assayed. The apoptosis rates of Leydig cells were detected by TUNEL. The visceral fat was isolated and weighed, and the Lee's index calculated. All the above indexes were recorded and compared among different age groups.The aging rats showed a significant decrease in the levels of serum tT and TSI ([1.26 +/- 0.65] ng/ml and [0.07 +/- 0.65] ng/mIU) as compared with the young rats ([3.24 +/- 0.38] ng/ml and [0.21 +/- 0.01] ng/mIU) (P0.01). Obvious differences were found in the morphology of Leydig cells among different age groups. The T secretion of Leydig cells at 24, 48 and 72 h in aging rats was markedly decreased (P0.05) while their TUNEL positive rate remarkably increased in the aging rats (17.36% +/- 1.31%) compared with the young ones (7.02% +/- 1.05%) (P0.05). There were statistically significant differences between the young and aging rats in all the biochemical parameters including IRG, HDL, LDL, TG, TC and visceral fat content (P0.05), except the levels of serum Glu, INS and LP (P0.05).The serum T level and secreting capacity of Leydig cells are significantly lower in aging rats than in young ones, and the metabolic parameters undergo regular changes with the decreasing level of serum T. The reduction of testosterone in aging male rats may be associated with the decreased secreting capacity and number of Leydig cells and declined function of the pituitary.

To investigate the incidence of late onset of hypogonadism (LOH) in males as well as the sensitivity, specificity and applicability of the androgen deficiency in aging males (ADAM) questionnaire and the aging males' symptoms (AMS) scale in Chinese males based on the community population data.A total of 1 498 old and middle-aged healthy males attended the ADAM and AMS investigations, of whom 434 received the measurement of the concentration of serum reproductive hormones, the positive rate of LOH screening, the rate of androgen deficiency, the clinical incidence of LOH, and the sensitivity and specificity of ADAM and AMS by tT and fT cut-off. The symptom evaluation cut-off value of the AMS score was optimized using the ROC curve.Although 5 of the subjects had previously received irregular testosterone supplement, none of them was given or actively sought for androgen therapy at the time of the investigation. Among the 40 - 69 years old males, the mean positive rates of LOH screening by ADAM and AMS were 80.77% and 32.34%, and the mean androgen deficiency rates obtained by tT and fT cut-off were 14.02% and 43.69%. The mean clinical incidences of LOH in the ADMA- and AMS-positive subjects were 37.85% and 15.42%. According to the fT cut-off, the sensitivities of ADAM and AMS were 86.63% and 35.29%, and their specificities were 24.48% and 63.49%, respectively. The symptom evaluation cut-off value of the AMS score optimized by the ROC curve was 19.5.The sensitivity and specificity of ADAM and AMS in the Chinese population are basically consistent with the results of most studies abroad, while the positive rate of LOH screening, the rate of androgen deficiency and the clinical incidence of LOH obviously higher in the former than those reported in other studies. Both ADAM and AMS are applicable to the Chinese population. The former is advantageous for its high sensitivity, time saving and easy operation, and therefore suitable for screening LOH, while the latter can be used for monitoring therapeutic efficacy.

To investigate the changes of serum reproductive hormones with male aging and to compare the differences in the hormone levels among different age groups or between township and rural males of the same age group.Using cluster and stratified sampling, we recruited 434 healthy old and middle-aged (40-69 years) males, 198 from the township and 236 from the rural communities. We determined the concentrations of serum total testosterone (tT), luteinizing hormone (LH) and sex hormone binding globulin (SHBG), free testosterone (fT), bio-available testosterone (Bio-T), and obtained the testosterone secretion index (TSI) and free testosterone index (fTI). Meanwhile, we included fifty-nine 20-39 years old males from the same communities in a control group.With the increase of age, the serum tT levels did not change significantly, while the levels of serum LH and SHBG increased, and those of fT, Bio-T, TSI and fTI decreased gradually. Statistically significant differences were found among the four different age groups in all the parameters of reproductive hormones (P0.01), except in the serum tT level (P0.05). The serum tT level was not significantly correlated with aging and LH (P0.05). Serum LH and SHBG had a marked positive correlation with aging, and SHBG with LH (P0.01), while fT, Bio-T, TSI and fTI were negatively correlated with aging and the LH level (P0.01). Serum LH, TSI and fTI showed statistical differences (P0.05), while fT and Bio-T exhibited extremely significant differences (P0.01) between the township and rural males in the 40 -49 yr group, and in the same age group, the increase rates of serum LH and SHBG and reduction rates of fT, Bio-T, TSI and fTI were higher in the rural men than in the township residents. However, the results were just the opposite in the 50 - 59 and 60 - 69 yr groups.The levels of serum LH, SHBG, fT, Bio-T, TSI and fTI changed with aging in a gradientmanner in the old and middle-aged males, but no significant changes were observed in the level of serum tT. There were statistical differences in many parameters of serum reproductive hormones among different age groups or between township and rural males.

To analyze the changes of seminal parameters in Chinese fertile men during the past 25 years.We collected semen samples from 5,834 fertile men in 14 different provinces (including Beijing) between 1980 and 2005 and retrospectively studied their seminal parameters, abstinence durations and total testis volumes by meta-analysis.Compared with the first 15 years, a significant decrease was observed in both sperm density and total number of sperm per ejaculate in the semen samples collected between 1996-2000 (P0.0001 and P0.05), but not obvious in those between 1996-2000 and after 2005 (P0.05). As for sperm motility, no time-related changes were noted (P0.05) except a reduction with the increase of age.There was a decline in sperm density and total number of sperm per ejaculate in Chinese fertile men over the past 25 years, although not significant in the latter 10 years since 1996, but with no time-related changes in sperm motility.

To study (CAG) n polymorphism of the androgen receptor (AR) gene in responders and non-responders of male volunteers who received testosterone undecanoate intramuscular injection for contraception and to explore the effect of the polymorphism on hormonal male contraception.Twenty-nine non-responders and 34 responders were enrolled in this study as a test and a control group respectively. The numbers of CAG sequence repeats were determined by PCR and DNA sequencing, and the effect of (CAG) n polymorphism on hormonal male contraception was analyzed.The means of CAG repeats of the test and the control group were 23.62 and 22.97, with no significant difference in between (P0.05). The short CAG repeats (nor = 22) constituted 51.7% in the test group and 50% in the control, while the long ones (n22) accounted for 48.3% and 50% , respectively. The short and the long group had a similar distribution. No association was found between CAG repeats and sperm concentration. With FSH0.2 IU/L, the probability of azoospermia in the long CAG repeat group was 1.5 times that of the short one.CAG repeats in the AR gene presented polymorphism in the subjects, with no significant difference between the responders and non-responders. Further investigation has yet to be performed into the relationship of hormonal male contraception with CAG repeats or other factors.

To investigate the changes and variation of serum antisperm antibodies (AsAb) from the men who received vasovasostomy 2 years after one of three methods of male sterilization, and evaluate the relationship between serum AsAb and pregnancy rate.Sperm reappearance, serum AsAb and pregnancy rate were investigated in 88 subjects who underwent reversal operation 2 years after one of three methods of male sterilization at 3, 6, 12 and 24 months post vasovasostomy.There was no significant difference in positive rate of AsAb among groups of vasovasostomized men post one of three methods of sterilization, and between groups before and post vasovasostomy and between groups with and without sperm reappearance, respectively. The positive rate, intensity (percentage of motile sperm coated with beads) and type of serum AsAb had no influences on pregnancy. No significant difference was observed in positive rate of AsAb between pregnant group and non-pregnant group. The pregnancy rate of the group in which sperm concentration was more than 20 x 10(6)/ml was apparently higher than that of the group in which sperm concentration was less than 20 x 10(6)/ml.The serum AsAb and their positive rate can not be eliminated or decreased by vasovasostomy post male sterilization. The serum AsAb has no influence on pregnancy. There is a close relationship between sperm quality and pregnancy post vasovasostomy.