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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。.

This multi-center, cross-sectional study investigated the association between serum testosterone (T) levels, serum sex hormone-binding globulin (SHBG) levels, and the risk of metabolic syndrome (MS) in 3332 adult Chinese men. The prevalence of MS was 34.7%, and men with MS had lower serum levels of total T (TT) and SHBG than those without MS (P < 0.001). There was no significant difference in serum free T (FT) levels between subjects with and without MS (P = 0.627). In logistic regression analysis, the association between MS and serum SHBG levels persisted after adjusting for age, body mass index (BMI), smoking and drinking status, and serum TT (odds ratio [OR] 0.962, 95% confidence interval [95% CI] 0.954−0.969, P< 0.01). However, the association between serum TT level and the risk of MS was weak after adjusting for age, BMI, SHBG level, and smoking and drinking status (OR 0.981, 95% CI 0.960−1.007). Our study reveals that both serum TT and SHBG levels, but not serum FT, are inversely associated with the prevalence of MS and that serum SHBG is an independent and dominant risk factor for MS.

Because of unavoidable complications of vasectomy, this study was undertaken to assess the efficacy and safety of male sterilization with a nonobstructive intravas device (IVD) implanted into the vas lumen by a mini-surgical method compared with no-scalpel vasectomy (NSV). IVDs were categorized into two types: IVD-B has a tail used for fixing to the vas deferens (fixed wing) whereas IVD-A does not. A multicenter prospective randomized controlled clinical trial was conducted in China. The study was comprised of 1459 male volunteers seeking vasectomy who were randomly assigned to the IVD-A (n = 487), IVD-B (n = 485) or NSV (n = 487) groups and underwent operation. Follow-up included visits at the 3 rd -6 th and 12 th postoperative months. The assessments of the subjects involved regular physical examinations (including general and andrological examinations) and semen analysis. The subjects' partners also underwent monitoring for pregnancy by monthly interviews regarding menstruation and if necessary, urine tests. There were no significant differences in pregnancy rates (0.65% for IVD-A, 0 for IVD-B and 0.21% for NSV) among the three groups (P > 0.05). The cumulative rates of complications at the 12 th postoperative month were zero, 0.9% and 1.7% in the three groups, respectively. In conclusion, IVD male sterilization exhibits a low risk of long-term adverse events and was found to be effective as a male sterilization method, similar to the NSV technique. IVD male sterilization is expected to be a novel contraceptive method.

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 investigate associations among occupational exposure to coke oven emissions (COEs), oxidative stress, cytogenotoxic effects, change in the metabolizing enzyme glutathione S-transferase (GST), and internal levels of polycyclic aromatic hydrocarbons (PAHs) in coke oven workers, we recruited 47 male coke oven workers and 31 male control subjects from a coke oven plant in northern China. We measured the levels of 1-hydroxypyrene (1-OHP) and 8-hydroxy-2 -deoxyguanosine (8-OHdG) in urine, micronucleated binucleated cells (BNMNs) in peripheral blood lymphocyte, and GST in serum. Our results showed that the group exposed to COEs had significantly increased levels of 1-OHP [median 5.7; interquartile range (IQR), 1.4-12.0 micromol/mol creatinine] compared with the control group (3; 0.5-6.4 micromol/mol creatinine). In addition, the median levels (IQR) of 8-OHdG, BNMNs, and GST were markedly increased in the exposed [1.9 (1.4-15.4) micromol/mol creatinine; 6 (2-8) per thousand ; 22.1 (14.9-31.2) U/L, respectively] compared with controls [1.3 (1.0-4.0) micromol/mol creatinine, 2 (0-4) per thousand; and 13.1 (9.5-16.7) U/L, respectively]. These results appeared to be modified by smoking. However, multivariate logistic regression analysis revealed that exposure to COEs had the highest odds ratio among variables analyzed and that smoking was not a significant confounder of the levels of studied biomarkers. Overall, the present findings suggest that COE exposure led to increased internal PAH burden, genetic damage, oxidative stress, and GST activity. The consequences of the changes in these biomarkers, such as risk of cancer, warrant further investigations.

Background Telomerase maintains telomere length and is considered to be necessary for the indefinite proliferation of human cells. Activation of telomerase plays a key role in the malignant transformation process. The aim of this study was to study the regulation of telomerase, and to explore the possibility of telomerase as a biomarker in squamous carcinogenesis of the esophagus. Methods Twenty-nine esophageal squamous cell carcinomas (ESCC) and its corresponding adjacent normal tissues, and 47 epithelial squamous dysplasia tissues were analyzed by the reverse transcriptase-polymerase chain reaction (RT-PCR) technique for the mRNA expression of three major telomerase subunits: human telomerase RNA (hTR), telomerase protein component 1 (TP1), and human telomerase reverse transcriptase (hTERT) and by telomeric repeat amplification protocol assay (TRAP) for telomerase activity. Results For the expression of hTR and TP1 mRNA, there were no significant differences among ESCC, dysplasia and normal tissues (P > 0.05). In contrast, hTERT mRNA expression was detected in 28 of 29 ESCC (96.6%), in 23 of 47 dysplasia (48.9%), and only in two of 29 normal tissues (7.5%). Telomerase activity was positive in 25 of 29 ESCC (86.2%), in 21 of 47 (44.7%) epithelial dysplasia tissues, and in none of normal tissue. All together, 95 of 105 cases (90.48%) were concordant for both results, i.e., telomerase activity positive and hTERT positive or telomerase activity negative and hTERT negative tissues, and telomerase activity correlated with hTERT mRNA expression (P

Cyclooxygenase-2 (cox-2) overexpression has been observed in several types of human cancers and has been implicated in carcinogenesis. To elucidate the role of cox-2 in esophageal carcinogenesis, we evaluated the expression of cox-2 in normal squamous epithelium squamous epithelial dysplasia (n=47), and squamous cell carcinoma of the esophagus (n=86) by immunohistochemistry, reverse transcription-PCR assay, and western blotting. A significant overexpression of cox-2 was observed in esophageal squamous dysplasia and squamous cell carcinoma compared with normal squamous epithelium. The immunoreactive score of cox-2 expression, an index determined by intensity and positivity of cox-2 staining, was 0.71 +/- 0.46 (mean +/- SD) in normal squamous esophagus, 2.19 +/- 1.79 in squamous epithelial dysplasia, and 2.67 +/- 1.77 in squamous cell carcinoma. The results of immunohistochemistry were confirmed by a reverse transcription-PCR assay and western blotting analysis. Cox-2 expression level was correlated with proliferation activity assessed by proliferating cell nuclear antigen (PCNA) index and MIB-1 index in dysplastic lesion (r=0.55, P0.01 with PCNA and r=0.72, P0.01 with MIB-1) and carcinoma (r=0.56, P0.01 with PCNA and r=0.72, P0.01 with MIB-1). Elevated cox-2 expression was associated with high p53 expression (p0.001) but not with clinicopathological features including age, sex, tumor size, histological grade, lymph node metastasis, and TNM stage. The results indicated that cox-2 may be involved in an early stage of squamous carcinogenesis of the esophagus, and that cox-2 overexpression was related to cell proliferation in esophageal squamous dysplasia and squamous cell carcinoma.

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 explore the effects of sperm chromatin structure abnormalities on the outcome of in vitro fertilization and embryo transfer (IVF-ET).Sperm DNA fragmentation and chromatin packaging defects were assessed in 136 couples undergoing IVF-ET because of infertility. The relationship between sperm DNA fragmentation, chromatin packaging defects and fertilization rate and clinical pregnancy was evaluated.Both sperm DNA fragmentation and chromatin packaging defect had a negative correlation with fertilization rate (r=-0.198, P0.05, and r=-0.389, P0.01, respectively). Both parameters were higher in couples who failed to achieve pregnancy than those who achieved clinical pregnancy (10.74% vs. 5.40%, P0.01 and 23.58% vs. 11.83%, P0.01, respectively).Abnormality of sperm chromatin structure is one of the reasons for IVF-ET failure. Examination of sperm chromatin structure is helpful in predicting the risk of IVF-ET failure and optimizing treatment of infertility.

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.

Because of the daily and life-long exposure to disinfection by-products formed during drinking water treatment, potential adverse human health risk of drinking water disinfection is of great concern. Toxicological studies have shown that drinking water treatment increases the genotoxicity of surface water. Drinking water treatment is comprised of different potabilization steps, which greatly influence the levels of genotoxic products in the surface water and thus may alter the toxicity and genotoxicity of surface water. The aim of the present study was to understand the influence of specific steps on toxicity and genotoxicity during the treatment of surface water in a water treatment plant using liquid chlorine as the disinfectant in China. An integrated approach of the comet and oxidative stress assays was used in the study, and the results showed that both the prechlorination and postchlorination steps increased DNA damage and oxidative stress caused by water extracts in human derived L-02 cells while the tube settling and filtration steps had the opposite effect. This research also highlighted the usefulness of an integrated approach of the comet and oxidative stress assays in evaluating the genotoxicity of surface water during drinking water treatment. Environ. Mol. Mutagen. 2010. © 2009 Wiley-Liss, Inc.

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.

To investigate the application of serum glutathione S-transferase (GST) and urinary 8-hydroxy-2-deoxyguanosine (8-OHdG) as the monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons (PAHs).47 male coke oven workers and 31 male control workers were investigated. Urinary 8-OHdG and serum GST were analyzed using high performance liquid chromatography (HPLC) with electrochemical detection and test kit. Urinary 1-hydroxypyrene (1-OHP) as internal exposure of PAHs was also determined simultaneously by alkaline hydrolysis and HPLC.The values of urinary 1-OHP, serum GST and urinary 8-OHdG were reported as median with interquartile range (P(25)-P(75)). Urinary 1-OHP [5.7 (1.4-12.0) micromol/mol Cr], serum GST [22.1 (14.9-31.2) U/ml], and urinary 8-OHdG [1.9 (1.4-15.4) micromol/mol Cr] in coke oven workers were significantly higher than in control workers [3.0 (0.5-6.4) micromol/mol Cr (P0.05), 13.1 (9.5-16.7) U/ml (P0.01), and 1.3 (1.0-4.0) micromol/mol Cr (P0.05) respectively]. Categorizing by smoking status, significant differences in urinary 1-OHP and serum GST were found only in smokers among coke oven workers compared to control workers (P0.01), and 8-OHdG levels only in non-smokers (P0.01). Additionally, there was significant correlation between urinary 1-OHP and serum GST activity (r(s) = 0.31, P0.01, n = 78). The multiple logistic regression analysis showed that coke oven workers were at the higher risk of having GST activities above 16.7 U/ml (OR = 13.2) and 8-OHdG levels above 1.8 micromol/mol creatinine (OR = 4.4). High body mass index was an independent factor to affect urinary 8-OHdG levels.The elevated serum GST activities and increased oxidative DNA damage were found in the coke oven workers. Occupational exposure and smoking interact on each other. Serum GST may be used as a biomarker for assessing the exposure of PAHs. Assay of urinary 8-OHdG may be useful for evaluating the risk of lung cancer in coke oven workers.

To explore the lipid peroxidation induced by deltamethrin (DM) in the cerebral cortex and hippocampus of rat.Wistar male rats were administrated with DM (daily dose was 3.125, 12.500 mg/kg respectively). The content of malondialdehyde (MDA) and the activity of total-superoxide dismutase (T-SOD, including Mn-SOD and CuZn-SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione peroxidase (GSH-Px) and glutathione reductase (GR) in cerebral cortex and hippocampus tissue were determined. The reduced glutathione (GSH) content and gamma-glutamylcysteine synthetase (gamma-GCS) activity in cytosolic fraction of cerebral cortex and hippocampus tissue was determined by reversed-phase high performance liquid chromatographic assay with o-phthalaldehyde pre-column derivation.(1) MDA content in cerebral cortex of the high dose group was significantly higher than those in the low dose group, and MDA content in hippocampus tissue of the high dose group was significantly higher than those in both the control and the low dose group after 5 d of DM exposure. (2) The activity of T-SOD and CuZn-SOD in cerebral cortex of both high and low dose group were significantly lower than that in the control group, and there was no effect on CAT activity in cerebral cortex (P0.01 or P0.05). (3) GSH content in cerebral cortex of the high dose group was significantly higher than that in control group (P0.05), and that in hippocampus tissue of high dose was significantly lower than that in both control and low dose group (P0.05). GR activity of low dose group in cerebral cortex was significantly lower than that in both control and high group [(11.80 +/- 5.15) vs (18.98 +/- 3.68), (17.35 +/- 2.47) U/mg pro] (P0.01). Gamma-GCS activity in hippocampus tissue of the high dose group was significantly lower than that in both control and low dose group [(1.75 +/- 0.60) vs (3.17 +/- 0.79), (2.72 +/- 0.75) nmol x mg pro(-1) x min(-1)] (P0.01). GR activity in hippocampus tissue of both high and low dose group was significantly lower than that in the control group [(21.63 +/- 4.92), (21.46 +/- 8.89) vs (31.22 +/- 6.97) U/mg pro] (P0.05).The oxidative stress in nerve tissue, which could be resulted from effect of DM on the activity of SOD, gamma-GCS and GR and GSH content, is one of the mechanisms of neuro-toxicity induced by DM; The decreased activity of gamma-GCS and GR may be the primary cause of DM-induced decrease in that GSH content in hippocampus tissue.

Background: The purpose of the present paper was to study the expression of cyclooxygenase-2 (COX-2) in normal squamous epithelium, squamous dysplasia and squamous cell carcinoma (SCC) of the esophagus, to elucidate the role of COX-2 in esophageal carcinogenesis, and to evaluate the in vitro effect and mechanism of a COX-2 inhibitor, NS-398, in inducing growth inhibition and apoptosis of human esophageal cancer cells. Methods: Biopsy specimens of esophageal dysplasia (n = 21), and surgical resections of SCC (n = 37) were compared with normal esophagus (n = 37) and analyzed by RT-PCR. Human esophageal cells were used for the study. Anti-proliferative effect was measured by MTT, apoptosis was determined by DNA fragmentation assay. Results: Marked COX-2 expression was shown in SCC and esophageal squamous dysplasia, and no marked COX-2 expression was observed in the normal squamous epithelium, respectively. NS-398 could inhibit esophageal cells growth in a dose-dependent manner, induce apoptosis, and elevate caspase-3 activity in vitro. Conclusions: This study provides evidence that COX-2 is upregulated in the majority of cases of squamous dysplasia and SCC of esophagus, and that NS-398 can inhibit growth and induce apoptosis via activating caspase-3 activity in vitro. These results suggest that selective inhibitors of COX-2 may be an effective preventive and therapeutic option for esophageal carcinoma.

Telomerase maintains telomere length and is considered to be necessary for the indefinite proliferation of human cells. Activation of telomerase plays a key role in the malignant transformation process. The aim of this study was to study the regulation of telomerase, and to explore the possibility of telomerase as a biomarker in squamous carcinogenesis of the esophagus.Twenty-nine esophageal squamous cell carcinomas (ESCC) and its corresponding adjacent normal tissues, and 47 epithelial squamous dysplasia tissues were analyzed by the reverse transcriptase-polymerase chain reaction (RT-PCR) technique for the mRNA expression of three major telomerase subunits: human telomerase RNA (hTR), telomerase protein component 1 (TP1), and human telomerase reverse transcriptase (hTERT) and by telomeric repeat amplification protocol assay (TRAP) for telomerase activity.For the expression of hTR and TP1 mRNA, there were no significant differences among ESCC, dysplasia and normal tissues (P0.05). In contrast, hTERT mRNA expression was detected in 28 of 29 ESCC (96.6%), in 23 of 47 dysplasia (48.9%), and only in two of 29 normal tissues (7.5%). Telomerase activity was positive in 25 of 29 ESCC (86.2%), in 21 of 47 (44.7%) epithelial dysplasia tissues, and in none of normal tissue. All together, 95 of 105 cases (90.48%) were concordant for both results, i.e., telomerase activity positive and hTERT positive or telomerase activity negative and hTERT negative tissues, and telomerase activity correlated with hTERT mRNA expression (P0.001).Higher telomerase activity and hTERT mRNA expression were shown during an early stage in the esophageal carcinogenesis. Activation of telomerase activity was strongly correlated with hTERT mRNA expression, suggesting hTERT is a major regulator of telomerase activity, and telomerase activation may play a critical role in esophageal carcinogenesis. Therefore, telomerase, especially hTERT can be used as a potential molecular biomarker of ESCC.

To investigate the expression of cyclooxygenase-2 (cox-2) protein in normal squamous epithelium, squamous dysplasia and squamous cell carcinoma of the esophagus, and to elucidate the role of cox-2 in esophageal carcinogenesis.Biopsy specimens of atypical esophageal dysplasia (n = 47) and surgical resection of squamous cell carcinoma (n = 86) were compared with normal esophageal specimens (n = 42) and the expression of cox-2 in those specimens was analyzed by immunohistochemistry and western blotting.A significant elevated cox-2 expression was shown in atypical esophageal squamous dysplasia and squamous cell carcinoma, as compared to that in normal esophageal squamous epithelium, with immunohistochemical stain scores of 2.67 +/- 1.77, 2.19 +/- 1.79 and 0.71 +/- 0.46, respectively. Results of western blotting analysis confirmed those obtained by immunohistochemistry. Cox-2 expression significantly correlated with proliferation activity assessed by proliferating cell nuclear antigen index in dysplastic and carcinomous lesions, respectively, and no such correlation could be found in normal esophageal mucosa. Elevated cox-2 expression was not associated with clinical-pathological features of esophageal squamous carcinoma, including age, gender, tumor size, histological grade, lymph node metastasis and clinical stage.Elevated expression of cox-2 in atypical squamous dysplasia and squamous cell carcinoma of the esophagus, which correlated with cell proliferation activity, indicated that cox-2 may be involved in the early stage of squamous carcinogenesis of the esophagus, and may be a target of prevention and treatment for esophageal squamous cell carcinoma.

To investigate the effect of X-ray repair cross complementing 1 (XRCC1) genetic polymorphisms on esophageal cancer risk, we determined XRCC1 polymorphisms at codon 194 (Arg→Trp) and codon 399 (Arg→Gln) in 135 patients with esophageal squamous cell carcinoma (ESCC) and 152 normal controls from hospitals. Although polymorphism at codon 194 was not associated with risk for ESCC, we found that the frequency of XRCC1 399 Gln/Gln genotype in ESCC patients (14.1%) was significantly higher than that in normal controls (3.3%), and that XRCC1 399 Gln/Gln genotype was associated with an increased risk of ESCC (odds ratio (OR)=5.15, 95% confidence interval (CI): 2.42–0.93). In addition, we found that the risk for smoker increased 4.2-fold than non-smokers in the 399 Gln/Gln genotype (OR=4.20, 95% CI: 2.37–7.44). These results suggest that XRCC1 399 Gln/Gln genotype may contribute to the risk of ESCC and modify risk associated with smoking.

To investigate the effects of scutellarin (Scu) on liver function after brain ischemia/reperfusion in Wistar rats.Rats were pretreated with Scu for 7 d and then subjected to a brain ischemia/reperfusion injury induced by a middle cerebral artery occlusion. The levels of nitric oxide (NO), xanthine oxidase (XOD), alanine transaminase (ALT), and aspartate aminotransferase (AST) in serum or liver tissues and the activities of antioxidant enzymes and cytochrome P450-dependent monooxygenases (CYPs) in liver tissues after brain ischemia/reperfusion were determined.In vehicle-treated rats, XOD, ALT, and AST activities (P0.01) in serum and the MDA level (P0.05) in liver tissues were all elevated but were significantly reduced (P0.05) by Scu pretreatment. The NO levels in serum and liver tissues were decreased (P0.01) dramatically in vehicle-treated rats and returned to the levels in the sham-operated animals when pretreated with Scu. SOD (P0.05) and GSH-PX (P0.01) activities in cytosol fraction were increased significantly by Scu pretreatment. Furthermore, a loss of CYP3A activity (P0.01), but no changes of CYP1A1, CYP1A2, and CYP2E1 activities in liver were observed after brain ischemia/reperfusion in rats. Scu had no effect on them.These results demonstrated that pretreatment with Scu could attenuate hepatocellular damage elicited by brain ischemia/reperfusion in rats and this protection is in major part by its antioxidant activity.

To study the relationship of human telomerase reverse transcriptase (hTRT) and malignant transformation of esophageal dysplasia.Telomerase activity and hTRT expression in esophageal dysplasia (n = 47), squamous cell carcinoma (n = 29) and normal esophagus (n = 11) were detected by telomeric repeat amplification protocol (TRAP) and in situ hybridization, respectively.Telomerase activity was detected in none of the 11 cases of normal esophageal tissues (0%) but in 21 of 47 cases (44.7%) of dysplasia, and in 25 of 29 cases (86.2%) of esophageal squamous cell carcinoma. There were statistically significant differences among the telomerase activity in normal esophagus, esophageal dysplasia, and in squamous cell carcinoma (chi(2) = 5.89, P0.05; chi(2) = 11.35, P0.01). hTRT mRNA was expressed in none of the 11 cases of normal esophageal tissues (0%) but in 23 of 47 cases (48.9%) of dysplasia, and in 24 of 29 cases (82.8%) of esophageal squamous cell carcinoma. There were statistically significant differences among the expression of hTRT mRNA in normal esophagus, esophageal dysplasia, and in squamous cell carcinoma (chi(2) = 6.99, P0.01; chi(2) = 7.32, P0.01). Significant correlation was found between the telomerase activity and the expression of hTRT mRNA (chi(2) = 57.91, P0.001).The mRNA expression of hTRT which paralleled to telomerase activity implied that there was a crucial role to play in regulating the activation of telomerase, and was closely related to the malignant transformation of esophageal dysplasia. hTRT might serve as a new, valuable biomarker to detect esophageal squamous cell carcinoma.

To investigate the effects of Ginkgo biloba extract (GbE) and tanshinone (Tan) on cytochrome P450 (CYP) isozymes and glutathione transferase (GT) in rats.Several CYP-dependent reactions were monitored in liver and kidney microsomes of male rats treated ig with GbE and Tan daily for 10 d. The activity of GT, the levels of malondialdehyde (MDA) and nitric oxide (NO) in the tissues were also determined.CYP1A1, 1A2, and 2B1 activities in the liver were all significantly increased (2-9.5 fold) by pretreatment with GbE or Tan (P0.01). An induction (1.4 fold) of CYP 2E1 activity was observed at the higher dose of GbE treatment (P0.01), but a reduction (1.9 fold) after Tan administration (P0.01). Whereas GbE could induce CYP3A (1.6 fold) (P0.01) but Tan had no effects. Furthermore, the activities of CYP 1A1 (5.6-8.9 fold) and 1A2 (2.6 fold) in the kidney were induced by GbE (P0.01). The activity of GT in rat liver receiving Tan was significantly increased (P0.05) and a dramatic reduction in the activity of GT in the kidney was observed in the GbE-treated group (P0.01). In addition, the GbE treatment markedly decreased the levels of MDA and NO in the tissues of rats (P0.01).The modulation of CYP isozymes by GbE and Tan may result in altered metabolism of coadministered drugs. In addition, GbE is an active antioxidant and nitric oxide inhibitor in vivo.

The anti-thrombin aptamer can recognize and interact with thrombin in high affinity. This interaction results in the change of the fluorescent anisotorpy in the solution of the aptamer labeled with fluorescence. We developed a new method based on the change of the fluorescent anisotropy intensity resulted from this interaction. The change in polarization was depended on the concentration of thrombin. The linear range of the determination is 0.1-100ng/ml. The detection limit for thrombin is 0.1 ng/ml. This method has been used to determine thrombin in human whole blood. Aptamer that change the fluorescence characteristics of attached fluorophore will be useful in noncompetitive fluorescence polarization assay.

Dear Editor, We are very glad to see that the Asian Journal of Andrology published a Special Issue on Semen Analysis in 21st Century Medicine, which well revealed some behind-the-scene controversies of the 5th edition of the WHO laboratory manual for the examination and processing of human semen 1. Three articles from the special issue, two on the reference values of semen parameters 2, 3 and another presenting the investigation results of 118 laboratories performing semen analysis in Mainland China 4, are very thought-provoking and we would like to share some of our views on these topics. First, we noticed that the lower limit of the reference values of semen parameters in the 5th edition of the manual is reduced. Can this reduction reflect a genuine decline in semen quality? We are doubtful on this point. The reasons for the reduction vary from the variations in population selection, and the biases in methodologies of measurement and data processing to the recommended application of 'stricter criteria'. It has been a long time since the change in semen quality of healthy men become a controversial issue throughout the world. There is no definite conclusion on the trend of overall semen quality, owing to the differences in study design, definition of normal men, specimen collection methods, methods used for measurement of sperm and for statistical analysis, selection biases, measurement errors, regional differences, and other factors such as age, years spent in semen collection, the period of abstinence, etc. In addition, researches on change of semen quality are only retrospective analyses. Various insurmountable biases and confounding factors also make the conclusion poorly reliable. Although careful efforts were made to reduce the biases in some studies, discrepancies in results still remained. With the publication of the 5th edition of the WHO manual, more concerns and debates would be aroused. So far, it is not yet known whether the reduced lower limit of the reference values can indicate the decline in semen quality, but strictly evidence-based study design, standardization of measurement criteria and quality control (QC) of semen analysis would explain the reduction in the lower limit. Second, we think it is necessary to establish the reference intervals for values of semen parameters based on a fertile population using the time-to-pregnancy method in China. Semen quality is commonly taken as a surrogate indicator of male fertility, reproductive toxicology, epidemiology and pregnancy risk assessments. Reference intervals for values of semen parameters from a fertile population and the standardization of procedures for the examination of human semen given in the 5th edition of the manual provide a realistic basis for medical professionals all over the world, especially in China, where more than one-fifth of the world population inhabits. China is in urgent need of reference values of semen parameters derived from fertile Chinese men, which calls for a prospective, multi-center and nation-wide population-based study. Such a large-scale study will provide evidence for the prognosis of fertility, diagnosis of infertility and impact of environmental factors on human existence. It will also be helpful for the State Food and Drug Administration of China to establish medical guidelines for male fertility regulation. Third, internal and external QC of semen analysis need to be further improved. Semen analysis is used widely and has a significant role in the assessment of male fertility. There have been reports on the absence of standardization on semen analysis 5, 6, 7, 8 and obvious differences among laboratories 9, 10, 11, and some researchers have called for a comprehensive QC 12, 13, 14, 15, 16. The lack of consistency and standardization 17 incurs difficulties for clinicians to interpret or compare the results of semen analysis from different laboratories. Therefore, improvements in the following aspects would help improve the quality of semen analysis and ensuring the reliability of the results: using accredited and standard measurement, implementing internal QC and external QC. Finally, the criterion for determining an eligible donor in China should be revised accordingly to elevate the present poor eligibility after comprehensive investigations. The current quality requirements for an eligible donor's semen as per the Essential Criterion and Technical Specification of the Human Sperm Bank, issued by the Ministry of Health of China, are based on the 4th edition manual. With the reduction in certain semen parameters such as semen concentration, motility and semen volume in the 5th edition, we believe that studies on the reference intervals for values of semen parameters from fertile men must be carried out before any modification of the criteria is made in China. Such expanded studies would lead experts to conduct extensive discussions and would thus provide informative suggestions for policy makers before making any modification in the criteria of the human sperm bank in China.