Purpose: We investigated the expression and the exact role of leptin under hypoxic conditions in the human testis. Materials and Methods: Five testes from patients treated with orchiectomy for prostate cancer were used to construct an in vitro hypoxic culture system for human testicular tissue. Immunohistochemistry was performed to analyze leptin protein expression. Leptin, leptin receptor and HIF-1 alpha mRNA were examined by quantitative reverse transcriptase-polymerase chain reaction. Serum and seminal plasma leptin, gonadal hormones and semen parameters were evaluated in 10 healthy donors and 42 infertile patients with varicocele before and after surgery. Results: The viability of in vitro cultured testicular tissue was well maintained within 48 hours based on the results of morphological analysis, cell number and cell specific mRNAs. Immunohistochemistry demonstrated that leptin was mainly expressed in seminiferous tubules. Interestingly the optical density of leptin, leptin mRNA and HIF-1 alpha mRNA was significantly increased under hypoxia. Leptin mRNA and HIF-1 alpha mRNA correlated positively (R-s = 0.843, p < 0.01). In the clinical study the concentration of seminal plasma leptin before varicocelectomy was markedly higher in patients (mean +/- SD 3.01 +/- 1.23 ng/l, p < 0.01). It was highest in the grade 3 group (mean 3.95 +/- 1.37 ng/l, p < 0.01) and significantly decreased in the 6-month postoperative group (2.35 +/- 0.78, p < 0.05). Furthermore, negative correlations were observed between seminal plasma leptin and the sperm concentration (R-s = -0.187, p < 0.05), and progressive motility (R-s = -0.234, p < 0.05). Conclusions: Leptin expression was induced under hypoxia in the human testis, probably via the HIF-1 alpha related response pathway. Seminal plasma leptin closely correlated with varicocele related spermatogenesis dysfunction. It might effectively reflect the testicular hypoxic environment.

INTRODUCTION:The premature ejaculation diagnostic tool (PEDT) was developed to standardize the diagnosis of PE and has been applied in many countries. However, a linguistic validation of the Chinese version of PEDT does not exist.AIMS:This study aims to undertake the Chinese validation of the PEDT and to evaluate its association with self-estimated intravaginal ejaculatory latency time (IELT) and clinical expert diagnosis of PE.METHODS:A Chinese version of PEDT was confirmed by andrologist and bilingual linguist. Participants were recruited among seven different communities of Shanghai from 2011 to 2012, and their information regarding self-reported PE, self-estimated IELT, expert diagnosis of PE, and PEDT scores were collected.MAIN OUTCOME MEASURES:Validity of the PEDT and its association with clinical expert diagnosis of PE and self-estimated IELT were analyzed.RESULTS:A total of 143 patients without PE (mean age 55.11 ± 7.65 years) and 100 men with PE (mean age 53.07 ± 8.08 years) were enrolled for validation. Of the patients in PE group, the number of men reporting self-estimated IELTs of ≤1, 1-2, and >2 minutes were 34 (34.0%), 22 (22.0%), and 44 (44.0%), respectively. The Cronbach's alpha score (α = 0.77) showed adequate internal consistency, and the test-retest correlation coefficients of each item (r ≥ 0.70, P < 0.001) indicated excellent stability over time. The frequency of agreement showed that there was excellent concordance between PEDT diagnosis and clinician diagnosis when the PEDT scores ≥11. An adequate correlation was found between total PEDT score and self-estimated IELT (ρ = -0.396, P < 0.001), and sensitivity and specificity analyses suggested a score of ≤8 indicated no time-defined PE (self-estimated IELT ≤1 minute).CONCLUSIONS:The Chinese version of PEDT is valid in screening the presence of PE among Chinese men. The PEDT showed an adequate negative correlation with self-estimated IELT and an excellent concordance with clinician diagnosis of PE.

Purpose: We investigated whether the sperm protamine-1/2 mRNA ratio and DNA fragmentation index are reliable biomarkers in patients with clinical varicocele. Materials and Methods: We performed a prospective study in 42 subfertile patients with left clinical varicocele and 10 normozoospermic healthy donors with proven fertility. All patients and female partners were seen 3 and 6 months after varicocelectomy for fertility examination. Real-time quantitative reverse transcriptase-polymerase chain reaction and SCSA (R) were performed to analyze the sperm protamine-1/2 mRNA ratio and DNA fragmentation index. Results: The protamine-1/2 mRNA ratio and DNA fragmentation index were significantly higher in the preoperative group than in the control group (p < 0.01). After varicocelectomy in the pregnant group the protamine-1/2 mRNA ratio recovered to the normal value and the DNA fragmentation index was significantly lower than in the preoperative group (p < 0.01). In the nonpregnant group the protamine-1/2 mRNA ratio and DNA fragmentation index did not differ (p > 0.05). However, significant differences were present preoperatively according to varicocele severity in the protamine-1/2 mRNA ratio and DNA fragmentation index (p < 0.05 and < 0.01, respectively) with the highest values in the grade 3 group. Nevertheless, postoperatively no difference in varicocele severity was noted (p > 0.05). The protamine-1/2 mRNA ratio was strongly related to preoperative and postoperative sperm concentration (R-s -0.238, p < 0.01), progressive motility (R-s -0.327, p < 0.01), total motility (R-s -0.206, p < 0.05) and DNA fragmentation index (R-s 0.293, p < 0.01) except for normal morphology (Rs -0.064, p > 0.05). Conclusions: The sperm protamine-1/2 mRNA ratio and DNA fragmentation index can effectively be used to evaluate male fertility. Male infertility due to varicocele may be associated with protamine deficiency and sperm DNA damage. The post-varicocelectomy protamine-1/2 mRNA ratio and DNA fragmentation index are associated with the post-varicocelectomy pregnancy rate.

Objective To study the relationship between the level of prostate specific antigen (PSA)and levels of sex hormonessuch asfollicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), estradiol (E2)in aging males in the Shanghai. Methods Using multi-stage sampling method, 1 720 cases of 40 to 80-year-old male resident were recruitedfrom the general population of Shanghai The levels of serum prostate-specific antigen (PSA) and levels of four sex hormones (FSH, LH, T, E2) were respectively measured in the sampling population. Results Of 1 720 people, 1 489 were enrolled in the study due to their compliant and comprehensive data.Their average age was 61.88±10 years old, average level of PSA 1.29±1.06, levels of sex hormone FSH(7.97±6.35)mIU/ml, LH(4.39±.73)mIU/ml, T(4.47±2.58)ng/ml, and E2(9.99±16.51)pg/ml, there is no significant correlation between PSA and FSH levels (r = 0.002, P > 0.05); There were no significant correlations between PSA level and LH level (r=-0.008, P >0.05), between PSA level and T level (r =-0.008, P >0.05), between PSA level and E2 level (r =-0.019, P >0.05). Conclusion No relationship between PSA and four sex hormones was found in aging males.

STUDY QUESTION:Could the protamine-1 to protamine-2 mRNA ratio serve as a biomarker to estimate the fertilizing capacity of sperm from men taking part in an IVF/ICSI programme? SUMMARY ANSWER:The protamine mRNA ratio clearly discriminates between fertile and subfertile men and sperm with a normal protamine mRNA ratio exhibit a higher fertilizing capacity in IVF/ICSI. WHAT IS KNOWN ALREADY:Aberrant sperm protamine ratios are associated with male factor infertility and mRNA ratio is comparable with protein ratio (due to transcriptional stop in elongating spermatids). STUDY DESIGN, SIZE, DURATION:The study population was drawn from subfertile men, whose female partners participated in IVF or ICSI programmes between September 2010 and February 2012. Normozoospermic healthy volunteers served as controls. Sperm cells were lysed, mRNA extracted, reverse transcribed and subjected to real-time quantitative PCR using specific primer pairs for protamine-1 and protamine-2. Relative protamine-1 and protamine-2 mRNA levels were analysed with the Mann-Whitney U-test (two-tailed). PARTICIPANTS/MATERIALS, SETTING, METHODS:Quantitative RT-PCR for protamines 1 and 2 has been performed in ejaculates from 32 normozoospermic volunteers (control, University Clinic Giessen, Germany) and 306 patients, whose female partners took part in an IVF (n = 76; University Clinic Hamburg, Germany and Shanghai Jiaotong University, China) or an ICSI (n = 230; University Clinic Munich, Germany and Kinderwunschzentrum Wiesbaden, Germany) programme. MAIN RESULTS AND THE ROLE OF CHANCE:The sperm protamine mRNA ratio in normozoospermic men (0.98 ± 0.3) differed significantly from that of ICSI patients (Munich 0.81 ± 0.1; Wiesbaden 0.78 ± 0.2; P < 0.001), while processed samples obtained from IVF patients revealed a normal protamine mRNA ratio (Hamburg 1.0 ± 0.07; Shanghai 1.0 ± 0.54). Normal protamine mRNA ratios were associated with a significantly higher total motile sperm count and a significantly higher percentage of progressively motile sperm. Sperm with a normal protamine mRNA ratio revealed a higher fertilization capacity (fc) in both IVF (53.6% of patients with fc > 80%; P = 0.017) and ICSI (65.1% of patients with fc > 70%; P = 0.028). LIMITATIONS, REASONS FOR CAUTION:The protamine mRNA ratio in an individual sperm cell used for ICSI may be different from the overall value obtained from a semen aliquot. WIDER IMPLICATIONS OF THE FINDINGS:Data are in line with current literature and suggest the protamine mRNA ratio as a diagnostic marker to estimate the fertilizing capacity of sperm. STUDY FUNDING:The German Research Foundation (DFG) to K.S., W.W. and A.P. (STE 892/9-2), as well as to A.S. and H.C.O. (SP721/1-3). COMPETING INTEREST(S):None.

Objective To evaluate the vascular endothelial function of patients with erectile dysfunction using Peripheral Arterial Tone (PAT), and analyze the related factors to the vascular endothelial dysfunction. Methods The endothelial function of 87 ED patients was measured by EndoPAT2000. The test was carried out on two separate finger tips. The endothelial function was evaluated by PAT ratio of the finger tip. And ED patients were investigated by physical examination, lipid metabolism, fasting plasma glucose, sexual hormones (testosterone[T] 、estradiol[E2]、prolactin[P]) and International index of erectile function 5(IIEF-5). The clinical diagnostic criteria of metabolic syndrome (MS) was in accordance with the 2005 International Diabetes Federation (IDF2005) issued by the international academic community. Results Average age of 87 ED patients was 33.8±7.2 years-old, including 52 with endothelial dysfunction and 35 with normal endothelial function. Fasting plasma glucose (FPG), waist circumference and weight levels of patients in vascular endothelial dysfunction group were significantly higher than the patients in non-vascular endothelial dysfunction group (P<0.05), and total cholesterol(TC) and triglyceride (TG) were also significantly higher (P<0.01), while high density lipoprotein (HDL) was significantly lower ( P<0.05), but blood pressure, low density lipoprotein (LDL), T, E2 and PRL had no difference ( P>0.05). Secondly, the prevalence of MS in vascular endothelial dysfunction with ED group was 42.3%, which was significantly higher than 20% in non-vascular endothelial dysfunction with ED group (P<0.05). Secondly, compared with normal endothelial function, the patients with endothelial dysfunc-tion were much severer (P<0.05). Conclusion The prevalence of MS in ED patients with vascular endothelial dysfunction was significantly higher. Central obesity, hyperlipidemia and diabetes mellitus were the major risk fac-tors for endothelial dysfunction. And erectile function in patients with endothelial dysfunction might be even worse.

OBJECTIVE To ascertain the epididymal sperm nuclear maturity in obstructive azoospermia (OA) patients. METHODS A total of 81 infertile males from andriatry clinic of Renji hospital were selected, including 32 OA patients (OA group) and 49 asthenospermia patients (asthenospermia group). Another 32 fertile males were recruited for the control group. All semen samples of the asthenospermia and control groups underwent computer aided semen analysis (CASA). For the OA group, sperm specimens were collected by percutaneous epididymal sperm aspiration (PESA) and semen analyses completed by manual method. Further acridine orange test and aniline blue stain assay were performed. The results were processed with SPSS 15.0. RESULTS No statistically significant differences existed in semen volume and sperm density between the asthenospermia and control groups (both P > 0.05). The OA group was significantly lower than the control group in sperm density (P < 0.01). The asthenospermia and OA groups were also significantly lower than the control group in sperm motility (both P < 0.01). As to the percentage of grade a + b sperm, the asthenospermia group was also significantly lower than the control group (P < 0.01). Compared with the control group (81% ± 9%, 85% ± 8%), the negative rate of acridine orange test and aniline blue assay of OA (57% ± 20%, 64% ± 20%) and asthenospermia group (62% ± 19%, 67% ± 15%) were statistically lower (all P < 0.05). CONCLUSIONS The sperm nuclear maturity is lower in the asthenospermia and OA groups than that in the control group. Male fertility should be further improved.

Objective To detect the microsurgery treatment effect for male infertility patients with varicocele including the sperm DNA fragmentation index (DFI) and the level of reactive oxygen species (ROS) in seminal plasma, moreover, explore their correlation. Methods One hundred and twenty left varicocele male infertility with oligospermia and asthenospermia were treated with microscopic ligation of internal spermatic vein in Renji hospital from January 2011 to December 2012. The number of subclinical varicocele (SVC), clinical varicocele I (VcI), clinical varicocele II(VcII) and clinical varicocele III(VcIII) were 30 respectively. Meanwhile, 30 semen samples with normal range of parameters were selected as the control group. Their spouse had been naturally conceived within one year. Semen routine parameters, the sperm DFI and the level of ROS in seminal plasma in each group were detected. Furthermore, the spouse pregnancy condition was followed-up. Results The level of ROS in SVC group had no difference with that of the control group (P>0.05), while the sperm DFI was increased (P<0.05), both of them in VcI group were higher than those of the control group (P<0.05)and in VcII and VcIII groups much significantly higher (P<0.01). The sperm DFI was positively correlated with the level of ROS in seminal plasma (r=0.683, P<0.01). The level of ROS and sperm DFI had no difference (P>0.05) in SVC group after microsurgery, whereas, the level of ROS in VcI group was lower (P<0.05) and sperm DFI was not changed (P<0.05). For VcII and VcIII groups, the levels of ROS were significantly decreased (P<0.01)after microsurgery, meanwhile, the sperm DFI in VcII group was declined after 3 months(P<0.05), that of VcIII group was 6 months. The level of ROS and sperm DFI in pregnancy group were lower than that of those whose spouses were not pregnant (P<0.05). Conclusion Vc II and Vc III might contribute to the high level of ROS in seminal plasma and the damage of sperm DNA, whereas, microsurgery will bring benefit for the condition. Furthermore, it had great importance for the examination of the sperm DFI to predict the pregnancy outcome.

Objective To investigate the morbidity and risk factors of erectile dysfunction (ED) of middle-aged and geratic men in shanghai for improving diagnosis, treatment and prevention of ED. Methods Males in seven districts of Shanghai were recruited in the study, and sample size was determined according to a formula of N=400 × (Q/P). A total of 2 000males whose age were from 40 to 80 years old finished the survey. Results After removing the invalid questionnaire, 1 591valid questionaires were acquired. The mean age of 1 591 males was (61.9 ±10.1) years. About 893 of them (56.1%) had no sexual life. Among 698 males with sexual life, 275 were definitely diagnosed by the self-reported symptoms and course standard of ED, which accounted for 39.4% of males with sexual life and 17. 3% of the total samples. After rejecting 599 cases of IIEF-5<5, 782 cases whose IIEF-5 score was lower than 22 were suspected of ED. Age, prostatitis and drinking were all related to the morbidity of ED. Conclusion The morbidity of ED of middle-aged and geratic men who still had sexual life in shanghai is 17.3%. Age, the history of prostatitis and drinking are related to the morbidity of ED.

Objective: To study significance of the related biochemical indicator (Fasting blood-glucose, Triglyceride, Alanine aminotransferase, Creatinine) in aging males with ED. Methods: Total of 1591 men over 40 years old in Shanghai urban area, selected by multi-stage sampling methods, had been investigated by the questionnaire of erectile dysfunction and serum biochemical indicator assays. According to the score of International Index of Erectile Function (IIEF-5), 992 sexually active men in the overall population were divided into mild ED group, moderate ED grourp, severe ED group and normal group. The differences of the biochemical indictor levels among 4 groups, and the correlation between ED severity and levels of biochemical indicators were comparatively analyzed. Results: In 992 men, incidence of ED accounted for 78.83%, and no significant differences were found in the mean levels of ALT and Cr, between ED groups and the normal group, however a significant difference (P<0.01) in the mean level of FBG was found between severe/moderate ED group and mild ED/normal group. For the mean level of TG, there was a significant difference (P<0.01)between mild ED group and normal group, and there was also a significant difference(P<0.05) between moderate ED group and normal group. Moreover, the mean level of TG in mild ED goup was significantly lower (P<0.05) than moderate ED group, and the mean levels of TG in moderate ED group was significantly lower (P<0.05) than that of severe ED group. Conclusion: Hyperglycemia is an important risk factor for ocurrence of severe(moderate) ED in aging males. No correlation was found between hypertriglyceridemia and the morbidity of ED in aging males, but high level of serum TG in aging males with ED might aggravate the progression of ED.