Target gene panel method versus whole-exome sequencing in detection of idiopathic hypogonadotropic hypogonadism in males / 中华男科学杂志

Objective@#To compare the efficiency of the target gene panel method and whole-exome sequencing (WES) in detecting idiopathic hypogonadotropic hypogonadism (IHH), and select a more suitable gene detection method.@*METHODS@#We selected 24 genes closely related to the molecular pathogenesis of IHH to make up the gene panel, detected the mutation sites in 73 patients with IHH using the panel method, and verified the results of sequencing with the Sanger method. Using the key words "idiopathic hypogonadotropic hypogonadism", we searched databases for relevant literature, calculated the positive rate of IHH detected by WES and compared it with that detected with the panel method.@*RESULTS@#Of the 73 cases of IHH detected with the panel method, 7 were found with pathogenic mutations, including 2 cases of FGFR1, 2 cases of CHD7, 2 cases of KISS1R, and 1 case of NR5A1 mutation. Sanger sequencing showed that the positive rate of the panel method was 9.7%. Of the 1 336 articles retrieved, 5 met the inclusion criteria and were included, in which WES revealed a positive rate of about 30%.@*CONCLUSIONS@#For detection of the diseases with clear mutated genes, the panel method is relatively inexpensive and has a high sequencing depth, while for detection of the diseases with complicated genetic patterns and unclear mutated genes, WES is more efficient. Further studies are needed for choice of the two methods for different purpose of detection./.

Correlation between CFTR 5T polymorphisms and the risk of congenital bilateral absence of the vas deferens / 中华男科学杂志

Objective@#To investigate the association between the 5T site polymorphism of the cystic fibrosis transmembrane conductance regulator (CFTR) gene and the risk of congenital bilateral absence of the vas deferens (CBAVD).@*METHODS@#This case-control study included 40 male patients with isolated CBAVD in the experimental group and 104 healthy men as controls. We used the Sanger sequencing method to encode the CFTR gene intron 9 (TG) m-n(T) and type the haplotypes, followed by a review and meta-analysis of the data obtained from the experiment and relevant literature from the PubMed, Web of science, Medline, CNKI and an exploration of the correlation between 5T mutation and the risk of CBAVD.@*RESULTS@#Sanger sequencing revealed 6 genotypes in the CBAVD patients, including TG11-5T, TG12-5T, TG13-5T, TG11-7T, TG12-7T and TG11-9T, and 7 in the healthy controls, which were TG11-5T, TG12-5T, TG10-7T, TG11-7T, TG12-7T, TG13-7T and TG11-9T. Compared with the controls, the CBAVD patients showed obviously increased rates of the TG12-5T haplotype (4.81% ［10/208］ vs 16.25% ［13/80］) and the TG13-5T haplotype (0% vs 7.5% ［6/80］), but no significant difference in the TG11-5T haplotype (1.92% ［4/208］ vs 2.50% ［2/80］). There was a statistically significant difference between the experimental and control groups in the TG12_13-5T haplotype (OR = 7.40, 95% CI: 4.83－11.34, P < 0.01). The TG12_13-5T haplotype was found to be highly correlated with CBAVD.@*CONCLUSIONS@#The haplotype of TG12_13-5T increases the risk of CBAVD in men, which has provided a theoretical basis for male reproduction.

Correlation of the single nucleotide polymorphism rs662 of PON1 with the risk of male infertility / 中华男科学杂志

<p><b>Objective</b>To investigate the correlation between the single nucleotide polymorphism (SNP) rs662 of the paraoxonase 1 gene (PON1) and the risk of male infertility.</p><p><b>METHODS</b>This case-control study included 403 male idiopathic infertility patients aged 29.00 ± 4.48 years in the case group and 329 normal fertile men aged 28.28 ± 4.08 years as healthy controls. We obtained DNA from the peripheral venous blood of the subjects, genotyped the SNP rs662 of PON1 by Sequenom MassArray, and analyzed the association between different genotypes of PON1 rs662 and male infertility using the logistic regression model.</p><p><b>RESULTS</b>Compared with the normal controls, the infertility patients showed a significantly increased level of follicle-stimulating hormone (FSH) (［16.30 ± 17.76］ vs ［4.72 ± 2.51］ U/L, P < 0.01) but a decreased percentage of progressively motile sperm (PMS) (［7.40 ± 14.17］ % vs ［41.93 ± 9.06］ %, P < 0.01) and sperm concentration (［2.74 ± 3.64］ vs ［75.83 ± 63.66］ ×10⁶/ml, P < 0.01). Statistically significant differences were not found in the other parameters between the two groups of subjects, nor in the correlation of male infertility with the heterozygous genotype GA versus the wild homozygous genotype GG (OR = 0.98, 95% CI: 0.63－1.53, P = 0.923) or the homozygous genotype AA versus the wild homozygous genotype GG (OR = 0.87, 95% CI: 0.56－1.34, P = 0.525).</p><p><b>CONCLUSIONS</b>The SNP rs662 of PON1 was not correlated with male infertility, which, however, needs to be confirmed by further studies with larger samples from a larger area.</p>

Correlation of the single nucleotide polymorphisms rs34349826 and rs6521 of the LHB gene with male infertility in Chinese men / 中华男科学杂志

<p><b>Objective</b>To study the relationship of the single nucleotide polymorphisms (SNP) rs34349826 (c.104 A>G) and rs6521 (c.114 C>G) of the luteinizing hormone beta-subunit (LHB) gene with male infertility in Chinese men.</p><p><b>METHODS</b>This case-control study included 405 males with primary infertility (the infertility group) and 424 normal fertile men (the control group), the former again divided into subgroups of oligospermia, severe oligozoospermia and azoospermia according to the sperm concentration. Clinical data were collected from all the subjects and genomic DNA obtained from their peripheral blood for genotyping rs34349826 and rs6521 of the LHB gene by Sequence MassArray. We analyzed the correlation of male infertility with the SNPs of the two loci using the logistic regression model as well as its association with their haplotype combination with the SHEsis online software.</p><p><b>RESULTS</b>There were statistically significant differences between the control and infertility groups in the semen volume (［3.51 ± 1.36］ vs ［3.74 ± 1.71］ ml, P <0.05), sperm concentration (［79.21 ± 61.60］ vs ［27.37 ± 30.80］ ×10⁶/ml, P <0.01), percentage of progressively motile sperm (［39.40 ± 9.64］ % vs ［11.90 ± 14.72］ %, P <0.01), and levels of serum luteinizing hormone (LH) (［3.29 ± 1.39］ vs ［6.25 ± 4.83］ IU/L, P <0.01) and follicle-stimulating hormone (FSH) (［4.56 ± 2.31］ vs ［15.64 ± 17.03］ IU/L, P <0.01). Logistic regression analysis revealed no correlation between male infertility and the genotypes of the rs34349826 and rs6521 loci of the LHB gene, and similar results were found in the subgroups of the infertile males. SHEsis analysis on the haplotypes of the rs34349826 and rs6521 loci showed the GG genotype combination to be a protective factor against male infertility.</p><p><b>CONCLUSIONS</b>The rs34349826 and rs6521 loci of the LHB gene were not related to male infertility, which can be further confirmed by larger-sample studies. The GG genotype combination is a protective factor against male infertility.</p>

Kallmann syndrome with deafness caused by SOX10 mutation: Advances in research / 中华男科学杂志

The transcription factor SOX10, as a major actor in the development of the neural crest, plays a key role in the maintenance of progenitor cell multipotency, lineage specification, and cell differentiation. Abnormalities of neural crest development in humans lead to a number of genetic diseases known as neurocristopathies or neural crest disorders. The mutation of SOX10 can cause Kallmann syndrome (KS), which is a clinically and genetically heterogeneous condition and defined by the association between anosmia and hypogonadotropic hypogonadism due to incomplete migration of neuroendocrine gonadotropin-releasing hormone (GnRH) cells along the olfactory, vomeronasal, and terminal nerves. Since then, there have been a number of related reports that mutation of SOX10 will lead to KS with deafness. This review focuses on the SOX10 gene and the advances in the diagnosis and genetic studies of KS with deafness caused by the mutatuin of SOX10.

Relationship between the FSHR Thr307Ala-Asn680Ser gene polymorphism and male infertility: A meta-analysis / 中华男科学杂志

Objective@#To assess the association of the FSHR Thr307Ala-Asn680Ser gene polymorphism with male infertility.@*METHODS@#We searched Pubmed, EMBASE, Web of Science, CNKI, and WANFANG databases for literature on the correlation of the FSHR Thr307Ala-Asn680Ser gene polymorphism with male infertility published from 2005 to the present time. According to the inclusion criteria, we included 12 epidemiological case-control studies and subjected them to a comprehensive analysis with the Stata11.0 software.@*RESULTS@#A total of 2 893 male infertility patients and 3 312 controls were involved in the 12 studies. The Thr307Ala (rs6165) gene polymorphism was shown to be a risk factor for male infertility among the three comparison models (homozygous comparison model, hybrid comparison model and dominant comparison model), with the pooled odds ratios (OR) of 1.26 (95% CI: 1.03－1.54, P = 0.023), 1.18 (95% CI: 1.03－1.36, P = 0.018), and 1.20 (95% CI: 1.05－1.37, P = 0.006), respectively. And the Asn680Ser(rs6166) polymorphism was a risk factor for male infertility in the homozygous comparison and recessive comparison models, with the pooled ORs of 1.24, (95% CI: 1.05－1.45, P = 0.009) and 1.20 (95% CI: 1.04－1.39, P = 0.013), respectively. Layered meta-analysis showed that in the homozygous comparison model, the Thr307Ala-Asn680Ser polymorphism is a risk factor for male infertility in the white population, with the OR of 1.37 (95% CI: 1.03－1.82, P = 0.003) and 1.21 (95% CI: 1.00－1.47, P = 0.048), respectively.@*CONCLUSIONS@#In the homozygous model (GG vs AA), the FSHRThr307Ala-Asn680Ser gene polymorphism might be a protective factor against male infertility.

Nucleotide polymorphism rs4880 of the SOD2 gene and the risk of male infertility / 中华男科学杂志

Objective@#To investigate the correlation of the single nucleotide polymorphism (SNP) rs4880 of the superoxide dismutase 2 (SOD2) gene with the risk of male infertility.@*METHODS@#This casecontrol study included 519 male patients with idiopathic infertility (aged 19－40 ［28.93±4.93］ years) in the case group and 338 fertile men (aged 19－40 ［28.40±4.25］ years) in the control group. We collected the clinical data, genotyped the SNP rs4880 of the SOD2 gene by Sequenom Mass Array, and analyzed the association of different genotypes with male infertility using the logistic regression model.@*RESULTS@#Statically significant differences were observed between the case and control groups in the level of folliclestimulating hormone (FSH) (［4.72±2.51］ vs ［15.65±17.24］ U/L, P< 0.01), the percentage of progressively mobile sperm (［9.12±13.5］ vs ［41.95±9.03］%, P< 0.01), and sperm concentration (［12.95±24.38］ vs ［72.88±45.60］ ×106/ml, P< 0.01), but not in other parameters. No correlation was found between male infertility and the heterozygous genotype TC (OR = 0.90, 95% CI: 0.65－1.25, P = 0.516) or the homozygous genotype CC (OR=1.49, 95% CI: 0.38－5.81, P = 0.566) as compared with the wild genotype TT, and similar results were obtained in the analysis of the subgroups.@*CONCLUSIONS@#The SNP rs4880 of the SOD2 gene was not correlated with male infertility, which, however, is to be supported by further studies with larger samples from more areas.

Single nucleotide polymorphism of the TP53 gene is not correlated with male infertility / 中华男科学杂志

Objective@#To investigate the correlation of the single nucleotide polymorphism (SNP) rs1042522 of the tumor protein p53 (TP53) gene with the risk of male infertility.@*METHODS@#This casecontrol study included 380 male patients with idiopathic infertility and 398 normal fertile men as controls from the Nanjing area. We genotyped the SNP rs1042522 of the TP53 gene by Sequence Mass Array and analyzed the correlation of the SNP with male infertility using the logistic regression model.@*RESULTS@#Compared with the normal controls, the patients with idiopathic infertility showed significantly decreased sperm concentration (［77.34±49.24］ vs ［13.13±24.96］ ×106/ml), percentage of progressively motile sperm (［42.55±9.57］ vs ［10.38±5.57］%), serum testosterone level (［14.07±5.36］ vs ［11.89±4.50］ nmol/L), and folliclestimulating hormone level (［16.80±18.20］ vs ［4.55±7.17］ U/L) (P < 0.05) but no statistically significant differences in other parameters. No correlation was observed between the SNP frequencies and male infertility and similar results were found in the subgroups of the cases.@*CONCLUSIONS@#SNP rs1042522 of the TP53 gene is not significantly correlated with the risk of male infertility.

CYP1A1 rs4646422 gene polymorphisms not correlated with male infertility in Chinese Han population / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the correlation of the CYP1A1 (rs4646422) gene polymorphisms with male infertility in the Chinese Han population.</p><p><b>METHODS</b>Using the Mass ARRAY iPLEX GOLD technique, we conducted a case-control study on theCYPlA1 (rs4646422) gene polymorphisms in 636 infertile males aged 21-49 years (case group) and 442 normal healthy men aged 23-47 years (control group) of the Chinese Han population. We analyzed the genotypes and allele frequencies in the two groups ofsubjects with the SPSS 20.0 software.</p><p><b>RESULTS</b>Compared with the wild homozygous genotype GG, the heterozygous genotype AG (OR = 1.06, 95% CI 0.81-1.38) and homozygous genotype AA (OR = 1.11, 95% CI 0.56-2.21) showed no correlation with male infertility, nor did the mutant allele A (OR = 1.06, 95% CI 0.85-1.32) in comparison with the wild allele G.</p><p><b>CONCLUSION</b>The CYP1A1 (rs4646422) gene polymorphisms might not be correlated with male infertility in the Chinese Han population.</p>

Androgen receptor gene polymorphisms and male infertility / 中华男科学杂志

The androgen receptor (AR), as a ligand-dependent transcription protein and a member of the steroid receptor superfamily widely present in the body, is involved in the adjustment of the function of androgens and plays an important role in spermatogenesis. Androgens participate in spermatogenesis by binding AR and initiating the expression of the target gene. The polymorphisms of the AR gene may change the structure of AR and affect its avidity of binding androgens, making their downstream target genes unable to transcribe proteins. With the development of DNA sequencing techniques, studies on the association of the polymorphisms of the AR gene with male infertility have become a hot topic.

FSH and FSHR gene polymorphisms and male infertility: An update / 中华男科学杂志

Follicle-stimulating hormone (FSH) is synthesized and secreted by the anterior pituitary, which binds to its receptors expressed on the membrane of Sertoli cells in the testis to bring about spermatogenesis. With the development of DNA sequencing technology, FSH SNPs rs10835638 and FSHR SNPs rs6165, rs6166, and rs1394205 were detected, which might directly affect the expression of FSH and activity of FSHR, resulting in male spermatogenic dysfunction. This review focuses on the relationship of FSH and FSHR gene polymorphisms with male infertility.

Therapeutic efficacy of compound Xuanju capsule on autoimmune prostatitis in rats: an experimental study / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of Compound Xuanju Capsule (CXC) on autoimmune prostatitis in rat models.</p><p><b>METHODS</b>Sixty healthy male Wistar rats were randomly divided into five groups of equal number: blank control, low-concentration purified prostate protein (low-conc PPP), low-conc PPP + CXC treatment, high-concentration PPP (hi-con PPP), and hi-conc PPP + CXC treatment. Autoimmune prostatitis models were established by intragastric administration of PPP solution at 15 mg/ml (low concentration) and 80 mg/ml, respectively. At 30 days after modeling, the rats in the blank control and low-conc and hi-conc PPP model groups were treated with normal saline, and those in the other two groups with CXC at a daily dose of 0.068 g/ml. At 30, 45, and 60 days, all the animals were sacrificed for observation of pathological changes in the prostate tissue and determination of the levels of IL-8, IL-10, and TNF-alpha in the serum.</p><p><b>RESULTS</b>Compared with the PPP models, the hi-conc PPP + CXC group showed significantly reduced levels of IL-8 and TNF-alpha in the serum at 45 days ([148.54 +/- 17.23] and [62.14 +/- 5.59] pg/ml vs [100.77 +/- 11.08] and [32.63 +/- 2.91] pg/ml, P < 0.05) and at 60 days ([143.69 +/- 17.28] and [59.38 +/- 5.50] pg/mlvs [95.77 +/-10.53] and [29.63 +/- 2.66] pg/ml, P < 0.05), and so did the low-cone PPP + CXC group at 45 days ([128.47 +/- 12.21] and [40.43 +/- 3.64] pg/ml vs [111.76 +/- 10.07] and [35.44 +/- 3.17] pg/ml, P < 0.05) and at 60 days ([131.07 +/- 10.93] and [43.34 +/- 3.91] pg/ml vs [97.46 +/- 8.75] and [30.44 +/- 2.75] pg/ml, P < 0.05). The serum level of IL-10 was remarkably elevated in the hi-cone PPP + CXC group as compared with that of the PPP models at 45 and 60 days ([189.14 +/- 16.78] and [184.14 +/- 15.89] pg/ml vs [230.48 +/- 29.96] and [248.48 +/- 31.03] pg/ml, P < 0.05), and so was it in low-cone PPP + CXC group ([223.14 +/- 17.87] and [224.14 +/- 17.93] pg/ml vs [231.42 +/- 23.18] and [249.42 +/- 24.97] pg/ml, P < 0.05). Pathological examination revealed morphological damages to the prostate tissue and infiltration of inflammatory cells in the model rats, but no obvious changes in the normal controls. At 15 days of treatment, the rats in the PPP + CXC group showed enlarged prostate glandular cavity, mild proliferation of epithelial cells, no obvious infiltration of inflammatory cells in the interstitial tissue, and a few visible fibrous tissues under the light microscope.</p><p><b>CONCLUSION</b>Compound Xuanju Capsule is efficacious on autoimmune prostatis in rats by reducing inflammatory changes in the prostate tissue and improving the expression of inflammatory factors.</p>

Detection of DPY19L2 gene mutation in a globozoospermia patient / 中华男科学杂志

<p><b>OBJECTIVE</b>Globozoospermia is mostly associated with homozygous deletion of the DPY19L2 gene. This study aimed to investigate the DPY19L2 gene mutation in a globozoospermia patient.</p><p><b>METHODS</b>We observed the sperm histomorphology of a patient with globozoospermia using Wright-Giemsa's staining and transmission electron microscopy, detected the mutation of the DPY19L2 gene by PCR amplification and DNA sequencing, and compared the findings with the sequences issued in the Genbank.</p><p><b>RESULTS</b>Wright-Giemsa's staining showed that all the spermatozoa were round-headed and lacked the acrosome, with the head nucleus darkly, fully and densely stained. Transmission electron microscopy revealed larger round sperm heads, with an even layer of unit membrane surrounding the nuclei and dispersed cytoplasmic vacuoles but no acrosomal structure. No DPY19L2 gene mutation was found by PCR amplification and DNA sequencing.</p><p><b>CONCLUSION</b>No homozygous mutation of the DPY19L2 gene was found in the globozoospermia patient, and therefore some other disease-causing genes might be involved.</p>

DPY19L2 gene and globozoospermia: an update / 中华男科学杂志

Globozoospermia is a severe teratozoospermia, and the cases with 100% round-headed sperm are rare clinically. Globozoospermia is generally characterized by absence or abnormality of acrosome, accompanied by round-headed sperm with deranged midpiece and tails. The acrosome normally contains the enzymes that enable sperm to fertilize oocytes, while defective sperm cannot independently fertilize oocytes either in vivo or in vitro, and therefore globozoospermia makes males infertile clinically. Recent studies show that the deletion of the DPY19L2 (dpy-19-like 2) gene is a major cause of globozoospermia. This paper updates the relationship between DPY19L2 and globozoospermia to provide some evidence for further studies on the gene diagnosis and molecular mechanisms of globozoospermia.

Cigarette smoking affects sperm plasma membrane integrity / 中华男科学杂志

<p><b>OBJECTIVE</b>To detect sperm plasma membrane integrity (PMI) of cigarette smoking infertile males using SYBR-14/ PI fluorescent staining and flow cytometry and investigate its clinical significance.</p><p><b>METHODS</b>We collected semen samples from 132 cigarette smoking infertile men and 70 normal fertile controls, the former divided into a heavy-smoker group (> 20 cigarettes a day, n = 68) and a light-smoker group (< or = 20 cigarettes a day, n = 64). We performed computer-assisted semen analysis of the semen samples, and determined sperm PMI by flow cytometry after rinsing with PBS and staining by SYBR-14/PI, the sperm with normal PMI indicated as the percentage of those emitting green fluorescence (SYBR-14+/PI- %), dead sperm as the percentage of those emitting red (SYBR-14-/PI+), and moribund sperm as the percentage of those emitting both green and red (SYBR-14+/PI+).</p><p><b>RESULTS</b>Both the heavy- and light-smoker groups showed significant differences in SYBR-14-/PI+ % and SYBR-14+/PI- % from the normal controls (P < 0.01 or P < 0.05). SYBR-14+/PI- % was remarkably lower, while SYBR-14-/PI+ % markedly higher in the heavy-smoker than in the light-smoker group (P < 0.05). There was a significant correlation between SYBR-14+/PI- % and sperm motility (r = 0.938, P = 0.000).</p><p><b>CONCLUSION</b>SYBR-14/PI fluorescent staining and flow cytometry analysis could quickly and exactly detect sperm PMI. Cigarette smoking reduces sperm PMI and consequently sperm motility, which might be an important factor of male infertility.</p>