STUDY QUESTION: Are ten-eleven-translocation (TET) 1-3 family enzymes involved in human spermatogenesis and do they impact male fertility? SUMMARY ANSWER: TET1, TET2 and TET3 are successively expressed at different stages of human spermatogenesis, and their expression levels associate with male fertility. WHAT IS KNOWN ALREADY: Spermatogenesis is a complex cell differentiation process accompanied by a drastic epigenetic remodeling. TET1-3 dioxygenases are essential for active DNA demethylation in the paternal pronucleus and in embryonic stem cells. STUDY DESIGN, SIZE, DURATION: Expression of TET1-3 mRNAs and proteinss and 5-hydroxymethylcytosine (5-hmC) proteins were analyzed in human testis tissues from men with obstructive azoospermia and exhibiting histologically normal spermatogenesis. Ejaculated spermatozoa from normozoospermic healthy volunteers, the 'controls' (TET1: n = 58; TET2-3: n = 63), and subfertile men who participated with their female partners in an ICSI-program, the 'patients' (TET1: n = 66; TET2-3: n = 64), were analyzed concerning the stored TET1-3 mRNAs, and the values were correlated to semen parameters and ICSI-outcomes. PARTICIPANTS/MATERIALS, SETTING, METHODS: Testis sections were used for in situ hybridization (ISH) and immunohistochemical (IHC) studies to determine TET1-3 mRNA and protein expression, and for immunofluorescence (IF) detection of 5-hmC. Sperm samples from controls were analyzed by western blot, immunocytochemistry (ICC) and RT-PCR concerning the presence of non-degraded TET1-3 protein and mRNA. Sperm samples from controls and patients were used for quantitative TET 1-3 mRNA analyses (reverse transcription-polymerase chain reaction) and for comparative statistical evaluations under consideration of semen parameters and ICSI-outcome (pregnancy). MAIN RESULTS AND THE ROLE OF CHANCE: During human spermatogenesis TET1-3 proteins are successively expressed: TET2 is expressed in the cytoplasm of late pachytene spermatocytes of Stage V, TET1 starts to be expressed in the nuclei of Step 1 round spermatids at Stage I, and TET3 starts to be expressed in the nuclei of Step 3 round spermatids at Stage III. Five-hmC appears only in Step 5 elongated spermatids. All three TETs are still detectable at the mRNA and protein level in sperm cells in considerable amounts. Control men generally exhibited higher levels of TET1-3 in sperm. TET1- and TET3-mRNA levels in sperm were significantly negatively correlated with age (P = 0.0025 and P = 0.0343) and positively correlated with progressive sperm motility (P = 0.0007 and P = 0.018). All TETs showed a significant association with sperm concentration (P < 0.03). Patients diagnosed with oligozoospermia and/or asthenozoospermia (TET1: n = 35; TET2-3: n = 32) showed significantly reduced TET1-3 in sperm in comparison to controls (P = 0.003, P = 0.041 and P = 0.028), but not compared with normozoospermic patients. Levels of TET3 in sperm was significantly associated with high-fertilization rates (P = 0.009). Concerning ICSI-outcome, the lowest levels of TET1-3 mRNAs in sperm were found in the non-pregnant group. Increased TET2 in sperm was significantly associated with pregnancy (P = 0.006). LIMITATIONS, REASONS FOR CAUTION: Our results concerning the association of them RNA level of TETs in ejaculated sperm cells to different fertility parameters are descriptive. Further studies clarifying the reasons for decreased TET1-3 levels in subfertile men and their effect on their sperm methylome are essential. WIDER IMPLICATIONS OF THE FINDINGS: The study gives a substantial indication that in human spermiogenesis, an active DNA demethylation process occurs with an involvement of TET enzymes, and that the level of TET1-3 expression is pivotal for male fertility. STUDY FUNDING: Research grant from the German Research Foundation (DFG) to U.S. (SCHA1531/1-1 and SCHA1531/2-1). COMPETING INTEREST(s): None.

genetic factors.Non-obstructive azoospermia (NOA) is characterized by the absence of sperm in the ejaculate secondary to the lack of sperm production in the testicle.Advances in genetic and genomic analysis have allowed for the rapid identification of candidate genes in NOA men.The aim of our study was to identify novel variants implicated in the pathogenesis of NOA through whole-exome sequencing (WES).METHODS: Homozygosity mapping with whole-exome sequencing was performed using the NimbleGen SeqCap EZ Exome V3 sample preparation kit and the Illumina HiSeq 2000 next-generation sequencing platform to sequence the exomes of two siblings with NOA from a consanguineous family.Genetic mutations that passed filter criteria were then validated using Sanger sequencing.We identified one candidate gene, Intraflagellar Transport Protein 140 (IFT140), based on its expression pattern in testes according to the Human Protein Atlas, as well the mutation 0 s damage prediction based on two online bioinformatic resources.We subsequently performed Sanger sequencing on 95 NOA men and 26 fertile men, all of whom were treated at our institution.RESULTS: WES identified 442 variants in the index patients.Four (0.9%) of these variants passed filter criteria, one of which was a six-nucleotide deletion (HG38, chr16:1,519,961 TCTTGGC>T) in exon 22 of IFT140.Family segregation showed that this mutation was homozygous in the two brothers who underwent WES, as well as in a third brother with NOA and their sister.This mutation was not identified in the 1000 Genome Project and was predicted to be deleterious by the bioinformatics tools, MutationTaster and PROVEAN.Sanger sequencing validated the mutation in this family and identified it as a heterozygous mutation in 1 (1.1%) of the 95 NOA men and none of the 26 fertile controls in our cohort.Pathologic correlation of this unrelated patient showed hypospermatogenesis with a clinical history significant for three failed cycles of in vitro fertilization (IVF).CONCLUSIONS: We have identified through WES a small indel mutation in IFT-140 as a novel, disease-causing variant.This mutation was detected in our cohort of NOA in addition to the index family assessed with a prevalence of 1.1%.Further studies are required to determine the role of IFT140 in spermatogenesis.

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.

OBJECTIVE:To explore the optimal treatment selection for treating varicocele (VC) male infertility patients accompanied with oligozoospermia or azoospermia of different Chinese medical syndrome types by comparing the efficacies of integrative medicine. METHODS:One hundred and twenty male infertility patients with VC accompanied with oligozoospermia or azoospermia were assigned to Chinese medical treatment group (A) and the surgical group (B), each consisting of three Chinese medical syndrome types, i.e., damp-heat stagnation syndrome (DHSS), Shen-deficiency blood stasis syndrome (SDBSS), and blood stasis stagnation syndrome (BSSS), 20 in each group. Corresponding Chinese medical treatment was administered to those in Group A, C, and E, while microscopic ligation of internal vena spermatic was administered to those in Group B, D, and F. The routine analysis of semen, biochemical analysis of seminal plasma, and serum sex hormones (prolactin, testosterone, follicle stimulating hormone, luteinizing hormone, and estradiol) were performed before treatment and by the end of the 24th week after treatment. RESULTS:Totally 18 patients' spouses were pregnant. Of them, 1 in Group A of DHSS (abbreviated as Group A), 3 in Group B of DHSS (abbreviated as Group B), 4 in Group A of SDB-SS (abbreviated as Group C), 5 in Group A of SDBSS (abbreviated as Group D), 1 in Group A of BSSS (abbreviated as Group E), and 4 in Group B of BSSS (abbreviated as Group F). After 24-week treatment, the sperm concentration, class a sperm percentage, class a + b sperm percentage, the motility rate, the seminal plasma of fructose density, and the seminal plasma neutral alpha-glucosidase were more significantly improved in Group B, C, D and F, when compared with the same group before treatment (P <0. 01, P <0. 05). There was no statistical difference in the aforesaid indices between before and after treatment in Group A and Group E (P >0.05). As for the improvement percentage of seminal routine indices, the difference of the seminal plasma of fructose density, and the difference of seminal plasma neutral alpha-glucosidase between before and after treatment in the same Chinese medical syndrome types, better effects were obtained in Group B than in Group A (P <0.01), and better effects were obtained in Group F than in Group E (P <0.01). There was no statistical difference between Group C and D (P >0.05). There was no statistical difference in the 5 items of sex hormones in each group between before and after treatment (P >0.05). CONCLUSIONS:Surgical treatment could effectively improve the semen quality for male infertility VC patients accompanied with oligozoospermia or azoospermia. Of them, Chinese medical treatment could be recommended to those of SDBSS who would not receive surgical treatment.

The experimental principles of electromagnetic induction and magnetic levitation experiments were introduced,and the instrument of electromagnetic induction was composed by the stepping motor,turntable,magnet,force sensor,photoelectric coupler,and so on.Using the instrument,the relationships among levitation force,magnetic pull and aluminum plate rotation speed were measured.The condition and phenomenon of occurring electromagnetic induction were showed.The comprehensibility to electromagnetic induction phenomena and electromagnetic induction law for the students was enhanced.

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.

精索静脉曲张(varicocele,VC)是生殖系统血管性疾病,由于静脉回流受阻或瓣膜失效血液返流等异常因素导致精索静脉的伸长、扩张及迂曲.VC是导致男性不育的重要病因之一,在男性人群中发病率为15％～20％,男性不育中VC发病率为25％～40％,通过手术治疗可以有效改善男性生育力[1].VC患者血清、睾丸组织和精液中都存在过量的活性氧(reactive oxygen spies,ROS),过量的ROS造成精子损伤是导致VC患者不育的重要原因[2].并且结扎精索静脉后,ROS含量明显降低[3].ROS对于有氧环境中生存的细胞而言,少量持续的活性氧有利于维持细胞的正常功能,但是过量的活性氧引起氧化应激(oxidative stress,OS),损伤机体的氧化防御系统,使精子也像其他需氧细胞一样,引起精子功能损伤,甚至凋亡[4].所以研究氧化应激在精索静脉曲张中致男性不育的发病机制中具有重要意义.

This paper presents a WCDMA capacity planning method based on indicators analysis and capacity consumption tests proposed by Shenzhen Branch of China Unicom in the study of a traditional wireless network capacity planning method.In accordance with this method,the 2011 capacity planning was implemented.The results prove this method can meet the needs of the market accurately and effectively,also provide a strong guarantee for further development of market.

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 relationship between young men with erectile dysfunction and metabolic syndrome and to find the major risk factors affecting young ED patients. Methods:120 young ED patients and 40 males with normal sexual life were investigated by physical examination,lipid metabolism,fasting plasma glucose and International index of erectile function 5.The clinical diagnostic criteria of metabolic syndrome was according to the 2005 International Diabetes Federation issued by the international academic community.The ED patients with non-MS and MS also examined Audio visual sexual stimulation to assess the erectile function indicators. Results:The prevalence rate of MS in ED group was 35% which was significantly higher than 15% in non-ED group(P<0.01).The index central obesity(waist circumference,weight,body mass index) and triglyceride(TG) levels of patients in ED group were significantly higher than the patients in non-ED group(P<0.01),while high density lipoprotein(HDL) was significantly lower(P<0.01),total cholesterol(TC) and low density lipoprotein(LDL) was higher(P<0.05),but blood pressure and fasting plasma glucose(FPG) was no significant difference(P>0.05).Secondly,compared with the ED group with non-MS,the patients with MS were much severer(P<0.05),and IIEF-5 score reduced with the increase number of the risk factors of MS.Finally,AVSS indicators of ED patients with MS were inferior to the non-MS group(P<0.05). Conclusions:The prevalence of MS in young ED patients is significantly high and the severity of ED is positively correlated with the number of MS risk factors,in which central obesity and hyperlipidemia may be the major risk factors.

男性生育力下降是近年来一直被关注的全球性问题,越来越多的研究发现在不育夫妇中,男性因素占到约50％的比例[1].肿瘤坏死因子-α(Tumor Necrosis Factor-α,TNF-α)是一种具有多功能活性的细胞因子,它具有促炎症反应、调节免疫系统和细胞凋亡等作用,与男性精液质量关系十分密切[2].精液中TNF-α由生殖细胞、睾丸间质巨噬细胞等多种细胞分泌,在正常生理状态下具有调节精子增殖分化、控制凋亡及促进睾酮分泌等多种生物学作用.但在病理状态下TNF-α却会导致男性精液质量降低、精子活力下降、抑制顶体反应、诱导精子凋亡及自身免疫性睾丸炎,严重影响男性生育力.为此,本文就TNF-α与男性精液质量之间的关系进行综述.

目的明确精索静脉曲张(varicocele, VC)男性不育患者通过显微外科手术治疗对于精浆活性氧(reactive oxygen spies, ROS)与精子DNA损伤指数(sperm DNA fragmentation index, DFI)的治疗效果并分析其相关性.方法选取2011年1月~2011年12月至上海交通大学医学院附属仁济医院泌尿男科就诊左VC伴少、弱精子症男性不育患者共120例进行显微外科精索内静脉结扎手术治疗,其中亚临床(s u b c l i n i c a l varicocele,SV C)、V CⅠ度(clinical varicocele,V cⅠ)、V CⅡ度(VcⅡ)、VCⅢ度(VcⅢ)患者各30例;另选取配偶于1年内自然受孕,且精液参数正常范围的精液标本30份作为对照组.检测各组精液常规参数,TBA法检测精浆ROS水平、精液吖啶橙试验检测精子DFI.观察比较各组VC患者术前、术后3月、术后6月女方怀孕情况、精液常规参数、精浆R O S 水平与精子D F I 情况,对数据进行统计学分析.结果 VC男性不育患者术前SVC组与对照组ROS水平无差异(P>0.05),精子DFI上升(P<0.05);VcI组ROS水平及精子DFI增高(P<0.05);VcII组及VcIII组ROS水平与精子DFI显著增高(P<0.01);精子DFI与精浆ROS水平呈正相关(r=0.683,P<0.01).SVC组术后精浆ROS水平与精子DFI较术前无差异(P>0.05);V cI组术后精浆ROS水平低于术前(P<0.05),精子DFI无差异(P>0.05);V cⅡ及VcⅢ组术后精浆ROS水平明显降低(P<0.01);VcII组术后3月精子DFI降低(P<0.05);VcⅢ组术后6月降低(P<0.05).手术后VC患者精浆ROS水平及精子DFI怀孕组低于未怀孕组(P<0.05).结论 VcⅡ及VcⅢ伴少、弱精子症男性不育患者精浆ROS水平上升且精子DFI较高,VC显微外科治疗可一定程度改善睾丸氧化应激环境及降低精子DNA损伤程度.其次,通过检测精子DFI对于预测妊娠结局具有重要意义.

It discusses a power-saving solution of BTS based on the carrier frequency's smart On/Off and proves its operational feasibility in existing network by test.Without adding any new hardware equipment,the smart On/Off feature of carrier frequency,which may be achieved by software functions,can help to save 6% of the power without any influence on the normal performance of the BTS or the KPI of existing network.The result indicates that the idle carrier frequencies may be powered on or off intelligently according to the change of traffic load on existing network to save the power supply of the BTSs.

The model of multi-agent middleware is designed based on the theory of the existing middleware technology and the multiagent system and the work flow of the model is discussed. Then the structure of mobile GIS based on the model of multi-agent middleware and the characteristics of mobile GIS are presented. The structure of mobile GIS included the client tier, the application tier, the service function tier and the data tier. In the end, the mobile inspection system is designed and developed that combined the mobile GIS architecture. The system will promote the development of mobile GIS and produce practiced in mobile inspection.

Changes of protein physicochemical indicators and quality of Portunus pelagicus were investigated, such as salt insoluble protein, ATPase activity, sulfhydryl content, moisture-loss ratios and the sensory evaluation, through spraying liquid nitrogen of -50℃, soaking in liquid of -50℃, air tunnel way freezing at -35℃, and fridge freezing under -20℃.The results indicated that the solubility of actomyosin, ATPase activities and -SH content of myofibrillar protein decreased during storage.Quick-freezing was able to reduce the denaturation of the protein.Soaking in liquid of -50℃ was the best way in maintaining the quality of P.pelagicus, while fridge freezing under -20℃was the worst.

DRP(Distribution Resource Planning)system makes the enterprises know about information from mar- keting channels timely and offers a kind of brand-new mode for the business and cooperation with the enterpri- ses'trade partners.In this paper,using the MVC design pattern and the iBATIS persistence framework,a DRP system based on.NET for small and medium enterprises is designed,and the main function modules,key tech- nologies and implementation of the system are dis- cussed.

This article analyzes the general technique of Spatial Data Mining based on the agent theory. After explaining the task principle of this structure, an agent frame for Spatial Data Mining has been designed in the practice of digital agriculture spatial information platform.