Genetic polymorphism of glutathione S-transferase T1 gene and susceptibility to idiopathic azoospermia or oligospermia in northwestern China

Aim： To investigate the association of glutathione S-transferase T1 （GSTT1） gene polymorphism in patients with idiopathic azoospermia or oligospermia in the northwestern China population. Methods： In the case-control study, GSTT1 genotypes were identified by multiplex polymerase chain reaction （PCR） with peripheral blood DNA samples from 78 patients with idiopathic azoospermia, 103 patients with idiopathic oligospermia and 156 age-matched controls with normal sperm concentration and motility, according to the criteria adapted from World Health Organization guidelines. All of the patients and controls were from northwestern China. Results： There is a significant association between GSTT1 null genotype with idiopathic azoospermia risk （odds ratio [OR]： 2.36, 95% confidence interval [CI]： 1.33-4.20, P = 0.003） or idiopathic oligospermia risk （OR： 2.00, 95% CI： 1.17-3.27, P = 0.010）. Conclusion： GSTT1 null genotype is a predisposing risk factor for sporadic idiopathic azoospermia or oligospermia in northwestern China. （Asian J Androl 2008 Mar; 10： 266-270）

Intervention of Xuduan Zhongzi Formula on spermatogenesis epididymal morphological changes in a mice model of oligospermia

Objective:To investigate the intervention of Xuduan Zhongzi Formula on the epididymis structure of model of oligospermia mice.Methods:Ten of the 45 male mice were labeled as the normal group,and the remaining 35 mice were injected with chloral hydrate for five consecutive days,and the randomly selected five mice were anesthetized with chloral hydrate.The left epididymis was removed,and a few sperm were found in the epididymis under the optical microscope,indicating successful construction of the model.They were randomly divided into three groups:the normal group,the L-carnitine group and the Xuduan group with 10 rats in each group.The the L-carnitine group and the Xuduan group were administrated with equivalent dose of human solvent,and the normal group and the model group were administrated with normal saline.After 8 weeks of intragastric administration,all experimental mice were killed,and the left epididymis was extracted for sperm detection,seminal plasma biochemical detection,HE staining,and electron microscopy.Results:In the model group,enlarged epididymal epithelial cells,vacuolar degeneration of primary and basal cells,and edema of interstitial cells and vascular dilation were found.Compared with the normal group,semen concentration,activity,SOD,A-glucosidase and fructose in the model group were significantly decreased,and the difference was statistically significant(P<0.01).After eight weeks of drug intragastric treatment,the semen concentration,activity,SOD,A-glucosidase and fructose in the Xuduan group were significantly increased compared with the model group,and the difference was statistically significant(P<0.01).Conclusion:Xuduan Zhongzi Formula can significantly improve the epididymal structure,the microenvironment of epididymal sperm maturation and the stability of epididymal epithelial structure of oligospermia model mice,creating conditions for sperm maturation in the epididymis.

Truncating PICK1 Variant Identified in Azoospermia Affected Mitochondrial Dysfunction in Knockout Mice

Objective The protein interacting with C kinase 1(PICK1)plays a critical role in vesicle trafficking,and its deficiency in sperm cells results in abnormal vesicle trafficking from Golgi to acrosome,which eventually disrupts acrosome formation and leads to male infertility.Methods An azoospermia sample was filtered,and the laboratory detection and clinical phenotype indicated typical azoospermia in the patient.We sequenced all of the exons in the PICK1 gene and found that there was a novel homozygous variant in the PICK1 gene,c.364delA(p.Lys122SerfsX8),and this protein structure truncating variant seriously affected the biological function.Then we constructed a PICK1 knockout mouse model using clustered regularly interspaced short palindromic repeat cutting technology(CRISPRc).Results The sperm from PICK1 knockout mice showed acrosome and nucleus abnormalities,as well as dysfunctional mitochondrial sheath formation.Both the total sperm and motility sperm counts were decreased in the PICK1 knockout mice compared to wild-type mice.Moreover,the mitochondrial dysfunction was verified in the mice.These defects in the male PICK1 knockout mice may have eventually led to complete infertility.Conclusion The c.364delA novel variant in the PICK1 gene associated with clinical infertility,and pathogenic variants in the PICK1 may cause azoospermia or asthenospermia by impairing mitochondrial function in both mice and humans.

A susceptibility locus rs7099208 is associated with non-obstructive azoospermia via reduction in the expression of FAM160B1

Non-obstructive azoospermia （NOA） is a severe defect in male reproductive health that occurs in 1% of adult men. In a previous study, we identified that rs7099208 is located within the last intron of FAM160B1 at 10q25.3. In this study, we analysed expression Quantitative Trait Loci （eQTL） of FAM16OB1, ABLIM1 and TRUB1, the three genes surrounding rs7099208. Only the expression level of FAM16OB1 was reduced for the homozygous alternate genotype （GG） of rs7099208, but not for the homozygous reference or heterozygous geno- types. FAM160B1 is predominantly expressed in human testes, where it is found in spermatocytes and round sper- matids. From 17 patients with NOA and five with obstructive azoospermia （OA）, immunohistochemistry revealed that expression of FAM160B1 is reduced, or undetectable in NOA patients, but not in OA cases or normal men. We conclude that rs7099208 is associated with NOA via a reduction in the expression of FAM160B1.

Preliminary study of letrozole use for improving spermatogenesis in non-obstructive azoospermia patients with normal serum FSH

We investigated whether letrozole （2.5 mg day-1） improves sperm count in non-obstructive azoospermia （NOA） patients. Four men were included in this study, and they had folliculo-stimulating hormone and other hormone levels within the normal range and no varicoceles or chromosomal aberrations. These four patients were administered letrozole for 3 months. Sperm count, testicular volume, gonadotropin, testosterone （T） and estradiol （E2） blood levels were assessed before, during and 1 week after the suspension of treatment. All patients showed spermatozoa in their ejaculate, increased gonadotropin and T levels and lower E2 levels （P〈0.05 in all cases）, when letrozole was administered. This suggests that letrozole treatment might improve sperm count in an NOA sub-population; however, more studies, including the proper controls, are needed to confirm its efficacy.

Testicular fine needle aspiration as a diagnostic tool in nonobstructive azoospermia

Aim： To report the fine needle aspiration cytology （FNAC） of the testes used as a diagnostic tool in non-obstructive azoospermic patients. Methods： One hundred and twenty-five non-obstructive azoospermic male candidates to intracytoplasmic sperm injetion （ICSI） were analysed for follicle stimulating hormone （FSH）, luteinizing hormone （LH）, testosterone and inhibin B plasma levels. They were classified into three groups on the basis of FNAC： 1） Sertoli cell-only syndrome （SCOS） （70）; 2） severe hypospermatogenesis （42）; and 3） maturation arrest （13）, Then, all men underwent testicular sperm extraction （TESE） for sperm recovery for ICSI. Results： Mature spermatozoa were detected by FNAC in 24 of 42 men with severe hypospermatogenesis and nine of 13 men with maturation arrest; while they were retrieved by TESE in 29 of 70 men with SCOS, 35 of 42 men with severe hypospermatogenesis （including the 24 by FNAC） and 10 of 13 men with maturation arrest （including the nine by FNAC）. The sensitivity and specificity of FNAC were 44.6 % and 100 %, respectively. There was no difference on testicular volume and hormonal parameters in men with and without sperm retrieved. Conclusion： These findings suggest that FNAC may be a simple and valid diagnostic parameter in non-obstructive azoospermic men and it may represent a valid positive prognostic parameter for sperm recovery at TESE, （Asian J Androl 2005 Sep; 7： 289-294）

Can inhibin-B predict the outcome of microsurgical epididymal sperm aspiration in patients with suspected primary obstructive azoospermia？

Aim： To evaluate whether inhibin-B can predict the outcome of a microsurgical epidymal sperm aspiration （MESA） procedure in patients with suspected primary obstructive azoospermia （OA） and if inhibin-B can replace testicular biopsy in the diagnostic work-up of these patients. Methods： Inhibin-B levels and testicular biopsy scores were related to the outcome of MESA in 43 patients with suspected primary OA. MESA was considered to be successful when epididymal sperm could be identified during the procedure. Results： Spermatozoa were present in the epididymal aspirate in 28 out of the 43 patients （65%）. lnhibin-B values were not significantly different in patients with successful or unsuccessful MESA. The modified Johnsen score, however, was significantly lower in patients with unsuccessful MESA （P = 0.003）. A rete testis obstruction or epididymal malfunctioning was found in 15% of patients with suspected primary OA, reflected by unsuccessful MESA despite normal inhibin-B levels and normal testicular histology. Conclusion： Inhibin-B cannot replace testicular biopsy as a diagnostic tool in the work-up of patients with suspected primary OA. Testicular biopsy is useful in identifying patients with spermatogenic arrest, who might have normal inhibin-B values.

Association of XRCC1 gene polymorphisms with idiopathic azoospermia in a Chinese population

Aim： To assess the possible role of genetic polymorphisms in DNA repair gene XRCC 1 （X-ray repair cross-comple- menting group l） during spermatogenesis by investigating the associations of one promoter polymorphism （T-77C） and two exonic polymorphisms （Argl94Trp and Arg399Gln） in XRCC1 gene with risk of idiopathic azoospermia in a Chinese population. Methods： The genotype and allele frequencies of three observed polymorphisms were examined by polymerase chain reaction-restriction fragment length polymorphism based on a Chinese population consisting of 171 idiopathic azoospermia subjects and 247 normal-spermatogenesis controls. Results： In our study, all the observed genotype frequencies were in agreement with Hardy-Weinberg equilibrium. The 399A （GA＋AA） allele frequency for idiopathic azoospermia subjects and controls was 0.216 and 0.269, respectively. Compared with GG genotype, the AA genotype of Arg399Gln showed a significant association with a decreased risk of idiopathic azoospermia （odds ratio = 0.315; 95% confidence interval = 0.12-0.86）. However, no significant differences were found between the cases and controls for T-77C and Arg194Trp polymorphisms. The major haplotypes of XRCC1 gene were TCG, TrG and TCA, whereas no haplotypes appeared to be significantly associated with idiopathic azoospermia based on the cutoff of P 〈 0.05. Conclusion： In a selected Chinese population, AA genotype of Arg399Gln appears to contribute to a decreased risk of idiopathic azoospermia, while we have not any evidence of involvement of XRCC1 T-77C and Arg194Trp polymorphisms in idiopathic azoospermia. （Asian J Androl 2007 Nov; 9： 843-848）

Comparison of sperm retrieval and reproductive outcome in azoospermic men with testicular failure and obstructive azoospermia treated for infertility

We assessed the rates of sperm retrieval and intracytoplasmic sperm injection outcomes, including the neonatal profile of infants conceived, in men with testicular failure. Three-hundred and sixty-five men with testicular failure who underwent micro-dissection testicular sperm extraction were included in this study. We compared their outcomes with 40 men with testicular failure who used donor sperm for injections due to failed retrieval, and 146 men with obstructive azoospermia who underwent percutaneous sperm retrieval. The retrieval rate in testicular failure was 41.4%, and the results were lower than the obstructed azoospermia （100%; adjusted odds ratio： 0.033; 95% Ch 0.007-0.164; P 〈 0.001）. Live birth rates after sperm injections were lower in men with testicular failure （19.9%） compared with donor sperm （37.5%; adjusted OR： 0.377 （95% Ch 0.233-0.609, P 〈 0.001）） and obstructive azoospermia （34.2%; adjusted OR： 0.403 （95% CI： 0.241-0.676, P= 0.001）. Newborn parameters of infants conceived were not significantly different among the groups. We concluded that the chances of obtaining sperm on retrieval and achieving a live birth after intracytoplasmic sperm injection （ICSI） are reduced in men with testicular failure. The profile of infants conceived after sperm injection does not seem to be negatively affected by testicular failure.

Prevalences of oligozoospermia and azoospermia in male partners of infertile couples from different parts of India

Aim： To determine whether there was any regional variation in the prevalence of azoospermia, oligozoospermia and mean sperm counts in male partners of infertile couples from different parts of India. Methods： Data on 16 714 semen samples analyzed over the past five years from six different laboratories located in five cities of India were collated and evaluated. Results： There was a regional variation in the prevalence of azoospermia. The prevalence of azoospermia was extremely high in Kurnool and Jodhpur （38.3% and 37.4%, respectively）. There was also a regional variation in the prevalence of oligozoospermia （51%） in Kurnool. There was no significant difference in the mean sperm counts in normospermic men. Conclusion： There is a regional variation in the prevalence of azoospermia and oligozoospermia in the male partners of infertile couples from different regions of India. The prevalence of azoospermia in Kumool and Jodhpur is higher than any other worldwide reported literature, Further studies need to be carded out to determine the cause of this. （Asian J Androl 2006 Jan; 8： 89-93）

Outcome of repeated micro-surgical testicular sperm extraction in patients with non-obstructive azoospermia

Aim： To evaluate the outcome of repetitive micro-surgical testicular sperm extraction （mTESE） attempts in non-obstructive azoospermia （NOA） cases, in relation to patients＇ initial testicular histology results. Methods： A total of 68 patients with NOA in whom mTESE had been performed in previous intracytoplasmic sperm injection （ICSI） attempts were reviewed. Results： Among the 68 patients with NOA, the first mTESE yielded mature sperm for ICSI in 44 （64%） （Sp^＋）, and failed in the remaining 24 （36%） （Sp^-）. Following their first trial, 24 patients decided to undergo a second mTESE. Of these 24 patients, no spermatozoa were obtained in 5 patients, and Sp^＋ but no fertilization/pregnancy were achieved in 19. In these 24 cases, mTESE was successively repeated for two （n = 24）, three （n = 4） and four （n = 1） times. The second attempt yielded mature sperm in 3/5 patients from the Sp group and 16/19 patients from the Sp^＋ group. At the third and fourth trials, 4/4 and 1/1 of the original Sp^＋ patients were Sp^＋ again, respectively. Distribution of main testicular histology included Sertoli cell-only syndrome （16%）, maturation arrest （22%）, hypospermatogenesis （21%） and focal spermatogenesis （41%）. Overall, in repetitive mTESE, 24/29 （82%） of the attempts were finally Sp^＋. Conclusion： Repeated mTESE in patients with NOA is a feasible option, yielding considerably high sperm recovery rate. In patients with NOA, mTESE may safely be repeated one or more times to increase sperm retrieval rate, as well as to increase the chance of retrieving fresh spermatozoa to enable ICSI.

Deletion or underexpression of the Y-chromosome genes CDY2 and HSFY is associated with maturation arrest in American men with nonobstructive azoospermia

Maturation arrest （MA） refers to failure of germ cell development leading to clinical nonobstructive azoospermia. Although the azoospermic factor （AZF） region of the human Y chromosome is clearly implicated in some cases, thus far very little is known about which individual Y-chromosome genes are important for complete male germ cell development. We sought to identify single genes on the Y chromosome that may be implicated in the pathogenesis of nonobstructive azoospermia associated with MA in the American population. Genotype-phenotype analysis of 132 men with Y-chromosome microdeletions was performed. Protein-coding genes associated with MA were identified by visual analysis of a genotype-phenotype map. Genes associated with MA were selected as those genes within a segment of the Y chromosome that, when completely or partially deleted, were always associated with MA and absence of retrievable testicular sperm. Expression of each identified gene transcript was then measured with quantitative RT-PCR in testicular tissue from separate cohorts of patients with idiopathic MA and obstructive azoospermia. Ten candidate genes for association with MA were identified within an 8.4-Mb segment of the Y chromosome overlapping the AZFb region. CDY2and HSFYwere the only identified genes for which differences in expression were observed between the MA and obstructive azoospermia cohorts. Men with obstructive azoospermia had 12-fold higher relative expression of CDY2transcript （1.33__.0.40 vs. 0.11＋_0.04; P=O.O003） and 16-fold higher expression of HSFYtranscript （0.78__.0.32 vs. 0.05_0.02; P=O.O005） compared to men with MA. CDY2 and HSFYwere also underexpressed in patients with Sertoli cell only syndrome. These data indicate that CDY2and HSFYare located within a segment of the Y chromosome that is important for sperm maturation, and are underexpressed in testicular tissue derived from men with MA. These observations suggest that impairments in CDY2 or HSFYexpression could be implicated in the pathogenesis of MA.

Uniform deletion junctions of complete azoospermia factor region c deletion in infertile men in Taiwan

Aim： To determine the deletion junctions of infertile men in Taiwan with azoospermia factor region c （AZFc） deletions and to evaluate the genotype/phenotype correlation. Methods： Genomic DNAs from 460 infertile men were examined. Bacterial artificial chromosome clones were used to verify the accuracy of polymerase chain reaction. Deletion junctions of the AZFc region were determined by analysis of sequence-tagged sites and gene-specific markers. Results： Complete AZFc deletions, including BPY2, CDY1 and DAZ genes, were identified in 24 men. The proximal breakpoints were clustered between sY1197 and sY1192, and the distal breakpoints were clustered between sY1054 and sYl125 in all but one of the 24 men. The testicular phenotypes of men with complete AZFc deletion varied from oligozoospermia, to hypospematogenesis, to maturation arrest. Conclusion： We identified a group of infertile men with uniform deletion junctions of AZFc in the Taiwan population. Despite this homogeneous genetic defect in the AZFc region, no clear genotypedphenotype correlation could be demonstrated. （Asian JAndrol 2006 Mar; 8： 205-211）

Clinical observation of loupe-assisted intussusception vasoepididymostomy in the treatment of obstructive azoospermia(analysis of 49 case reports)

To evaluate the clinical outcomes of loupe-assisted intussusception vasoepididymostomy(VE)in the treatment of epididymal obstructive azoospermia(EOA),we retrospectively analyzed data from 49 patients with EOA who underwent two-suture longitudinal intussusception vasoepididymostomy(LIVE)between 2000 and 2007.The data included the surgical method,postoperative motile sperm count per ejaculation,percentage of progressive motile sperm and patency and pregnancy outcomes.There were a total of 49 men undergoing scrotal exploration,and epididymal obstruction was found in all cases.Bilateral or unilateral anastomoses were performed in 40 and 6 men,respectively.The postoperative courses of 42 patients were followed up for more than 6 months,and the courses of 38 patients were followed up for more than 1 year.The overall patency and pregnancy rates were 71.4%and 26.3%,respectively.Moreover,progressive motile sperm was more frequently present in those patients who had undergone anastomosis at cauda than at corpus or caput.Pregnancy was achieved only in those patients who had undergone anastomosis at least on one side of the cauda epididymis.We think that the loupe-assisted method,with a lower overall cost and a simplified surgical procedure,can achieve satisfactory patency outcomes and pregnancy results.Data from this paper also suggest that paternity outcomes occur more frequently after anastomoses at cauda than at corpus or caput.

Surgical management of nonobstructive azoospermia

Nonobstructive azoospermia(NOA)is characterized by the complete absence of sperm in the ejaculate due to testicular failure.The evaluation and management of patients with NOA offer a challenge to the reproductive urologist.In the era of in vitro fertilization with intracytoplasmic sperm injection,surgical sperm extraction techniques can afford men with NOA biologic paternity.To provide a comprehensive review of surgical sperm retrieval approaches in the patient with NOA emphasizing complications,success rates and outcome optimization,a Medline search was conducted querying surgical approaches used to manage NOA.Four sperm extraction techniques are described including:testicular sperm aspiration,testicular sperm extraction,fine needle aspiration mapping and microdissection testicular sperm extraction.In addition,the roles for pre-extraction varicocelectomy and sperm cryopreservation are discussed.The management of NOA continues to evolve as newer tools become available.Several modalities of sperm acquisition exist.An understanding of their complications and success rates is fundamental to the treatment of NOA.

Testicular fine needle aspiration for sperm retrieval in non-obstructive azoospermia

Objective: To investigate the feasibility of obtaining mature spermatozoa for intracytoplasmic sperm injection (ICSI) by testicular fine needle aspiration (TEFNA) in men diagnosed non-obstructive azoospermia. Methods: TEFNA was performed in 121 patients with a mean of 15 punctures and aspirations from each testis with a #23 butterfly needle connected to a 20 mL syringe with an aspiration handle. Results: One hundred and twenty-one patients underwent 176 TEFNA cycles. Testicular sperm were recovered in 56.3 % (99/176) cycles from 57 % (69/121) of patients. The sperm recovery rate was 46.7 % (21/45) in patients with Sertoli cell-only syndrome, 45.7 % (16/35) in patients with maturation arrest, 96.1 % (25/26) in patients with hypospermatogenesis and 63.6 % (7/11) in patients of non-mosaic Klinefelter's syndrome as judged by testicular histology. No sperm were found in 3 cases with post-irradiation fibrosis and one, after resection and chemotherapy of unilateral testicular cancer. In 87 cycles of ICSI using the husbands' sperm, 591 mature oocytes were injected, 218 (36.9 %) were normally fertilized and 202 embryos developed; 178 were transferred in 62 cycles resulting in 26 pregnancies (41.9 %) with 44 gestational sacs (implantation rate: 24.7 %). Conclusion: TEFNA was an efficient, easy to learn, safe and well tolerated treatment in patients with non-obstructive azoospermia.

Oligospermia due to partial maturation arrest responds to low dose estrogen-testosterone combination therapy resulting in live-birth: a case report

A man having severe oligospermia, due to partial maturation arrest at spermatid stage, was given low dose estrogen-testosterone combination therapy for three months. His sperm count increased enormously, following which his wife conceived and delivered a healthy baby at term.

Identification of CFTR Gene Mutations in Chinese Patients with Congenital Obstructive Azoospermia

ve To analyze the frequency and hot spot of CFTR gene mutations in Chinese patients with congenital obstructive azoospermia

Network pharmacology research of kidney-invigorating and spleen-strengthening core drugs in treatment of oligospermia and asthenospermia

Objective:Exploring the material basis and mechanism of Kidney-invigorating and Spleenstrengthening method for treating oligospermia based on network pharmacology.Method:Select the effective elements and targets of Yishen Jianpi core drug pairs by searching the Chinese Medicine System Pharmacology Platform(TCMSP);select the disease targets of oligozoospermia by searching the human genetic information database;use R language software to select drugs and Disease common target,use Cytoscape software to build a drugdisease-target interaction network;build a common target protein interaction network(PPI)through the STRING platform;apply R language to the core drugs of Yishen Jianpi and oligospermia Enrichment analysis of GO and KEGG was performed on the common target.Result:A total of 97 effective active ingredients were obtained from Yishen Jianpi traditional Chinese medicine,and 46 possible targets for Yishen Jianpi traditional Chinese medicine to treat oligospermia were obtained.Combined with the results of KEGG enrichment analysis,it can be speculated that Yishen Jianpi traditional Chinese medicine improved male oligospermia The whole mechanism of action may be related to the regulation of human endogenous hormones.Conclusion:Through the research method of network pharmacology,the possible effect targets and pathways of Yishen Jianpi traditional Chinese medicine in the treatment of male oligospermia are predicted.

Frequency of Y chromosome microdeletions and chromosomal abnormalities in infertile Thai men with oligozoospermia and azoospermia

Aim： To investigate the possible causes of oligozoospermia and azoospermia in infertile Thai men, and to find the frequencies of Y chromosome microdeletions and cytogenetic abnormalities in this group. Methods： From June 2003 to November 2005, 50 azoospermic and 80 oligozoospermic men were enrolled in the study. A detailed history was taken for each man, followed by general and genital examinations. Y chromosome microdeletions were detected by multiplex polymerase chain reaction （PCR） using 11 gene-specific primers that covered all three regions of the azoospermic factor （AZFa, AZFb and AZFc）. Fifty men with normal semen analysis were also studied. Karyotyping was done with the standard G- and Q-banding. Serum concentrations of follicle stimulating hormone （FSH）, luteinizing hormone （LH）, prolactin （PRL） and testosterone were measured by electrochemiluminescence immunoassays （ECLIA）. Results： Azoospermia and oligozoospermia could be explained by previous orchitis in 22.3%, former bilateral cryptorchidism in 19.2%, abnormal karyotypes in 4.6% and Y chromosome microdeletions in 3.8% of the subjects. The most frequent deletions were in the AZFc region （50%）, followed by AZFb （33%） and AZFbc （17%）. No significant difference was detected in hormonal profiles of infertile men, with or without microdeletions. Conclusion： The frequencies of Y chromosome microdeletions and cytogenetic abnormalities in oligozoospermic and azoospermic Thai men are comparable with similarly infertile men from other Asian and Western countries.