Analysis of clinical phenotype and genotype of Chinese children with disorders of sex development / 中华儿科杂志

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.

Identification of a novel variant of SRD5A2 gene in a child featuring steroid 5α-reductase type 2 deficiency / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical characteristics and genetic basis of a child with 5α-reductase type 2 deficiency.@*METHODS@#Clinical data of the child was retrospectively analyzed. Targeted capture-next generation sequencing and Sanger sequencing were carried out to detect potential variants.@*RESULTS@#The patient's main features included micropenis and hypospadia. He was found to harbor compound heterozygous c.680G>A (p.R227Q) and c.3G>T (p.M1I) variants of the SRD5A2 gene. Among these, c.680G>A (p.R227Q) was inherited from his father and was a known pathogenic mutation, while c.3G>T (p.M1I) was inherited from his mother and was unreported previously.@*CONCLUSION@#The compound heterozygous variants of the SRD5A2 gene probably underlay the disease in this child, who was eventually diagnosed with 5α-reductase 2 deficiency.

Variant analysis on steroid 5-reductase type 2 deficiency caused by a novel SRD5A2 mutation / 中国当代儿科杂志

This article reported the clinical characteristics and SRD5A2 gene mutation pattern of a child with steroid 5-α reductase type 2 deficiency. The 2-month-old boy showed hypospadias and short penis shortly after birth. DNA was extracted from the peripheral blood of the child and his parents. The endocrine disease-related genes were captured and sequenced by high-throughput sequencing technology, and the family DNA samples were verified by Sanger sequencing. The results showed that c.680G>A(p.R227Q) and c.608G>A(p.G203D) compound heterozygous mutations existed in the SRD5A2 gene of the child. The c.680G>A mutation inherited from his father, which was a known pathogenic mutation. The c.608G>A mutation originated from his mother, which was a novel mutation discovered in this study. These results provide molecular evidence for the etiological diagnosis of the child and genetic counseling for the family, as well as extend the mutation spectrum of SRD5A2 gene.

Identification of three novel SRD5A2 mutations in Chinese patients with 5α-reductase 2 deficiency / 亚洲男科学杂志(英文版)

In this study, we investigated the genetics, clinical features, and therapeutic approach of 14 patients with 5α-reductase deficiency in China. Genotyping analysis was performed by direct sequencing of PCR products of the steroid 5α-reductase type 2 gene (SRD5A2). The 5α-reductase activities of three novel mutations were investigated by mutagenesis and an in vitro transfection assay. Most patients presented with a microphallus, variable degrees of hypospadias, and cryptorchidism. Eight of 14 patients (57.1%) were initially reared as females and changed their social gender from female to male after puberty. Nine mutations were identified in the 14 patients. p.G203S, p.Q6X, and p.R227Q were the most prevalent mutations. Three mutations (p.K35N, p.H162P, and p.Y136X) have not been reported previously. The nonsense mutation p.Y136X abolished enzymatic activity, whereas p.K35N and p.H162P retained partial enzymatic activity. Topical administration of dihydrotestosterone during infancy or early childhood combined with hypospadia repair surgery had good therapeutic results. In conclusion, we expand the mutation profile of SRD5A2 in the Chinese population. A rational clinical approach to this disorder requires early and accurate diagnosis, especially genetic diagnosis.

Karyotyping and analysis of 5α -reductase-2 gene mutation in 25 patients with hypospadias / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the karyotypes and SRD5A2 gene mutations in 25 patients with sporadic or familial hypospadias.</p><p><b>METHODS</b>The patients included 10 adults and 15 children, whose chromosomes were analyzed by G-banded karyotyping, and the SRD5A2 genes were sequenced.</p><p><b>RESULTS</b>Two patients were found to have an abnormal karyotype, while eight have carried compound heterozygous mutations of the SRD5A2 gene, which included 5 genotypes formed by 6 types of mutations, i.e., p.G203S/p.R227Q, p.R227Q/p.R246Q, p.Q6X/p.Q71X, p.L20P/p.G203S, and p.Q71X/p.R227Q. Mutations of the SRD5A2 gene were present in 32% (8/25) of all patients, 35% (8/23) in those with a normal karyotype, and 44.4% (8/18) in those with proximal type hypospadia. Bioinformatic analysis, literature review and pedigree analysis confirmed that all such mutations are pathogenic.</p><p><b>CONCLUSION</b>Chromosomal anomalies and mutations of the SRD5A2 gene are the main cause of hypospadias. Sequencing of the SRD5A2 gene may explain the etiology of nearly half of the patients with proximal type of hypospadas but a normal karyotype, which can facilitate genetic consulting.</p>

Expression of SRD5A1 and its prognostic role in prostate cancer: Analysis based on the data-mining of ONCOMINE / 中华男科学杂志

<p><b>Objective</b>To explore the expression of I-5α-reductase (SRD5A1)and its prognostic role in prostate cancer .</p><p><b>METHODS</b>Data about SRD5A1 were retrieved from the ONCOMINE database and the role of SRD5A1 in prostate cancer was analyzed.</p><p><b>RESULTS</b>Totally, 992 studies of different types relevant to the expression of SRD5A1 were identified in the ONCOMINE database. The SRD5A1 expression was statistically significant in 239 of the studies, overexpressed in 157 (11 in prostate cancer) and underexpressed in the other 82 (3 in prostate cancer). Eighteen of the studies, with 1 068 samples, addressed the expression of SRD5A1 in prostate cancer and normal tissues, which was significantly higher in the former than in the latter tissue (P<0.05). In 3 of the studies, the SRD5A1 expression was high in primary prostate cancer and increased with its metastasis (P<0.0 5). Two of the studies with prognostic data showed a higher rate of postoperative biochemical recurrence and a higher total mortality rate in the patients with a high than in those with a low expression of SRD5A1 (P<0.05).</p><p><b>CONCLUSIONS</b>SRD5A1 is highly expressed in prostate cancer, especially in metastatic and castration-resistant prostate cancer and its expression is associated with the prognosis of prostate cancer, which may be an important target of medication for prostate cancer.</p>

The association of 5-alpha reductase type 2 (SRD5A2) gene polymorphisms with prostate cancer in a Korean population

PURPOSE: Steroid 5-alpha reductase type 2 (SRD5A2) modifies testosterone to dihydrotestosterone (DHT) in the prostate. Single-nucleotide polymorphisms (SNPs) of the SRD5A2 gene might affect DHT. We sought to understand the relationship of SRD5A2 SNPs to prostate cancer in the Korean population. MATERIALS AND METHODS: Twenty-six common SNPs in the SRD5A2 gene were assessed in 272 prostate cancer cases and 173 controls. Single-locus analyses were conducted by using conditional logistic regression. Additionally, we performed a haplotype analysis for the SRD5A2 SNPs tested. RESULTS: Among the 20 SNPs and 4 haplotypes, there were no statistically significant results in the prostate cancer patients and the controls. In the logistic analysis of SRD5A2 polymorphisms with prostate-specific antigen (PSA) criteria, two SNPs (rs508562, rs11675297) and haplotype 1 displayed significant results (odds ratio [OR], 1.76; p=0.05; OR, 1.88-2.02; p=0.01-0.04; OR, 0.59; p=0.02, respectively). rs508562, rs11675297, rs2208532, and haplotype 1 (OR, 1.49; p=0.05; OR, 2.02; p=0.05; OR, 2.01; p=0.04; OR, 0.56-0.64, p=0.03-0.04, respectively) had significant associations with Gleason score. rs508562, rs11675297, and haplotype 1 (OR, 1.41-2.34; p=0.004-0.05; OR, 1.74-1.82; p=0.03-0.05; OR, 0.42-0.67; p=0.0005-0.03, respectively) were significantly associated with clinical stage. CONCLUSIONS: We conclude that there was no significant association between SRD5A2 SNPs and the risk of prostate cancer in the Korean population. However, we found that some SNPs and 1 haplotype influenced PSA level, Gleason score, and clinical stage.

Is the Expression of Androgen Receptor Protein Associated With the Length of AC Repeats in the Type III 5-alpha Reductase Gene in Prostate Cancer Patients?

PURPOSE: Type III 5-alpha reductase (SRD5A3; steroid 5-alpha reductase 3) may be associated with the progression of prostate cancer (PCa). The aim of our study was to determine whether the length of AC repeats in the SRD5A3 gene is associated with the risk of PCa and the expression of androgen receptor (AR) protein in Korean men. MATERIALS AND METHODS: We compared the length of AC repeats in the short tandem repeat (STR) region of the SRD5A3 gene in 68 PCa patients and 81 control subjects by genotyping. A total of 55 patients in the PCa group underwent radical prostatectomy. We evaluated the expression of AR protein by using Western blotting and tested the association between the type of AC repeats in the SRD5A3 gene and AR protein expression and clinical and pathologic parameters. RESULTS: The short type of STR had less than 21 copies of AC repeats in the SRD5A3 gene. The SS type (short and short type) of STR of the SRD5A3 gene was 2.2 times as likely to occur in PCa patients as in controls (odds ratio, 2.21; 95% confidence interval, 1.14 to 4.31; p=0.019). However, AC repeats of the SRD5A3 gene were not associated with AR protein expression or clinical or pathologic parameters in PCa samples. CONCLUSIONS: These results suggest that the short AC repeats of SRD5A3 polymorphism are associated with an increased risk of PCa. SRD5A3 polymorphism may contribute to a genetic predisposition for PCa.

V89L polymorphism of the testosterone 5-alpha-reductase II gene and prognostic factors of prostate cancer / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the association of V89L polymorphism of the SRD5A2 gene with the prognostic factors of prostate cancer (PCa).</p><p><b>METHODS</b>We identified the V89L polymorphic sites of the SRD5A2 gene after Rsa-1 restriction enzyme digestion, observed the distribution of V89L (VV, VL and LL) polymorphism in 112 PCa and 89 benign prostate hyperplasia (BPH) patients, and determined the association of V89L polymorphism with the age, free PSA (fPSA), total PSA (tPSA), fPSA/tPSA ratio, tumor stage and Gleason score of the PCa patients.</p><p><b>RESULTS</b>No statistically significant differences were found in the V89L polymorphism-induced genetic risk frequencies between the PCa and BPH groups (chi2 = 3. 606, df = 2, P = 0. 165), nor any significant correlation between the genotypes of VV and VL + LL and the differences in the fPSA, tPSA, fPSA/tPSA ratio, tumor stage, Gleason score and age of the PCa patients. VV and VL + LL showed no obvious association with the prognostic factors of PCa.</p><p><b>CONCLUSION</b>V89L polymorphism is not related with the prognosis of PCa, but may be indirectly associated with its risk.</p>

The Effect of Finasteride on Microvessel Density and Expression of Vascular Endothelial Growth Factor and 5alpha-Reductase in Prostatic Hyperplasia

PURPOSE: Vascular endothelial growth factor (VEGF), a potent stimulator of angiogenesis and microvessel density (MVD), which is an important indicator of neoangiogenesis, were independently evaluated to elucidate the mechanism of decreased bleeding observed in patients treated with finasteride, an inhibitor of 5alpha-reductase (5AR). We evaluated MVD and the expression of VEGF and 5AR type II in patients with benign prostatic hyperplasia (BPH) treated with finasteride. MATERIALS AND METHODS: The study included 61 patients undergoing transurethral prostatectomy (TURP) for BPH. Among these patients, 29 had well-preserved paraffin blocks, 13 of whom were given finasteride for a minimum of 3 weeks before surgery; the remaining 16 patients served as controls. MVD was calculated by counting the number of positively stained blood vessels on 5 random, high-power fields within the prostatic section. Expressions of VEGF and 5AR type II were analyzed with a confocal laser scanning microscope and an image analyzer. RESULTS: Prostatic MVD was significantly lower in the finasteride-treated group (p<0.05). The expression of VEGF and 5AR type II at the level of the prostatic glandular epithelium and stroma was not significantly different between the 2 groups. VEGF and 5AR type II were more strongly expressed in the epithelium of both groups than in stromal smooth cells (p<0.05). CONCLUSIONS: Finasteride treatment had no clear effect on the expression of VEGF or 5AR type II. It is possible, however, that finasteride improves blood loss after TURP and BPH-induced hematuria by reducing MVD. Further study on the mechanism of MVD reduction is needed.

Molecular diagnosis of 5alpha-reductase-2 gene mutation in two Indian families with male pseudohermaphroditism / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To identify the genotype of two Indians with male pseudohermaphroditism.</p><p><b>METHODS</b>Standard radioimmunoassay procedure was used for estimating hormonal levels. Conventional cytogenetic analysis was carried out for diagnosing the genetic sex in these subjects with genital ambiguity. Molecular analysis was carried out by standard polymerase chain reaction procedure using different sets of primers and reaction conditions specific for the 5alpha-reductase type 2 gene (SRD5A2) gene. Direct sequencing was carried out using the ABI Prism dye terminator sequencing kit and the ABI 310 sequencing apparatus.</p><p><b>RESULTS</b>We found an SRD5A2 gene mutation in exon 5, where arginine is substituted with glutamine (R246Q), in two males with pseudohermaphroditism and ambiguous genitalia from unrelated families. This is the first time this mutation has been reported in individuals from India.</p><p><b>CONCLUSION</b>Identification of the R246Q mutation of the SRD5A2 gene from two unrelated Indian families possibly extends the founder gene effect.</p>

Mutation analysis of SRD5A2 gene in patients with hypospadias / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore possible molecular mechanism of hypospadias and relationship of the mutation of SRD5A2 gene to hypospadias.</p><p><b>METHODS</b>96 blood samples from the patients with hypospadias were obtained and DNA was extracted from blood leukocytes. Polymerase chain reaction and direct sequencing were performed to analyze the coding regions of SRD5A2 gene.</p><p><b>RESULTS</b>8 mutations were detected from 14 cases, including 5 missense mutations, 1 synonymous mutation, 1 nonsense mutation and 1 frameshift mutation. The mutations are in 1st, 4th and 5th exon. Gln6stop, His232His, Phe234Leu and frameshift mutations are novel.</p><p><b>CONCLUSIONS</b>Exon 4 is a hot spot region of mutation within SRD5A2 gene, and about 10 percent of hypospadiac patients complicated with SRD5A2 dysfunction or deficiency.</p>

Association of polymorphisms in testosterone 5-alpha-reductase II genotype and prognosis factors of prostate cancer / 中华外科杂志

<p><b>OBJECTIVE</b>The correlation were studied between testosterone 5-alpha-reductase II (SRD5A2) gene polymorphisms and prognosis factors.</p><p><b>METHODS</b>V89L and A49T variants was identified with Mwo1 and Rsa1. The differences of V89L and A49T between cancer of prostate (CaP) and benign prostatic hyperplasia (BPH) were studied. In addition, we also researched the association of polymorphisms with age of onset, free prostate specific antigen (FPSA), total PSA (TPSA), FPSA/TPSA (F/T), Gleason score, and T stage in cancer group.</p><p><b>RESULTS</b>We found no differences of V89L and A49T polymorphisms between CaP and BPH. In CaP group the A49T variant was associated with lower age of onset (P = 0.03) and higher Gleason score (P = 0.015). There were no differences between VV and VL+LL polymorphisms with any of the characteristics studied. When the characteristics above were regarded as two-level discrete variable, there were no differences by A49T and V89Lvariants.</p><p><b>CONCLUSION</b>In CaP group, the AT+TT genotype was perhaps associated with poor prognosis. VL+LL genotype has no relation with prognosis.</p>

Diferenciação sexual do cérebro: do animal experimental ao homem / Sexual differentiation of the brain: from the experimental animals to human beings

As diferenças existentes entre machos e fêmeas na morfologia e nas modalidades de funcionamento de numerosos núcleos cerebrais são determinadas, em grande parte, pelo ambiente hormonal presente durante o desenvolvimento embrionário. Os hormônios esteróides, em modo particular a testosterona, desenvolvem um papel crucial na masculinização dos centros cerebrais que governarão a secreção de vários hormônios hipotalâmicos e hipofisários, o comportamento sexual e a capacidade de aprendizagem e de memorização do indivíduo adulto. Nos últimos anos, surgiram dois aspectos importantes em relação ao mecanismo de ação da testosterona: 1) sua conversão, diretamente em nível de células-alvo, em compostos em grau de amplificar ou diversificar sua ação; 2) a presença, no sistema nervoso central (em particular no hipotálamo), da 5-alfa-redutase e da aromatase, enzimas necessárias a tais transformações. A interação dos metabólitos ativos resultantes (respectivamente, diidrotestosterona e estradiol) com específicos receptores intracelulares induz a transcrição de genes que determinam a diferenciação, em sentido masculino, das estruturas cerebrais em via de desenvolvimento. Nesta breve revisão, descrevem-se os conhecimentos atuais das principais características das duas vias enzimáticas que medeiam efeitos de diferenciação dos androgênios sobre o cérebro embrionário. Particular atenção é direcionada à discussão dos dados experimentais, obtidos principalmente nos roedores, em relação aos efeitos dos estrogênios de origem androgênica sobre a diferenciação sexual do cérebro. Discute-se também o papel co-primário que o genótipo desenvolve na diferenciação dimórfica do cérebro. Por fim, aborda-se a possível influência dos metabólitos ativos da testosterona sobre a diferenciação sexual do cérebro humano à luz de algumas doenças que comportam desequilíbrio nos níveis normais ou no mecanismo de ação dos hormônios gonadais durante o desenvolvimento embrionário.

The 5alpha-reductase type 1, but not type 2, gene is expressed in anagen hairs plucked from the vertex area of the scalp of hirsute women and normal individuals

The aim of the present study was to determine the expression of the genes for type 1 (SDR5A1) and type 2 (SDR5A2) 5alpha-reductase isoenzymes in scalp hairs plucked from 33 hirsute patients (20 with polycystic ovary syndrome and 13 with idiopathic hirsutism) and compare it with that of 10 men and 15 normal women. SDR5A1 and SDR5A2 expression was estimated by RT-PCR using the gene of the ubiquitously expressed protein ß2-microglobulin as an internal control. The results are expressed as arbitrary units in relation to ß2-microglobulin absorbance (mean ± SEM). SDR5A2 expression was not detected in any hair samples analyzed in this study. No differences were found in SDR5A1 mRNA levels between men and normal women (0.78 ± 0.05 vs 0.74 ± 0.06, respectively). SDR5A1 gene expression in the cells of hair plucked from the scalp of normal women (0.85 ± 0.04) and of women with polycystic ovary syndrome (0.78 ± 0.05) and idiopathic hirsutism (0.80 ± 0.06) was also similar. These results indicate that SDR5A1 gene expression in the follicular keratinocytes from the vertex area of the scalp seems not to be related to the differences in hair growth observed between normal men and women and hirsute patients. Further studies are needed to investigate the expression of the 5alpha-reductase genes in other scalp follicular compartments such as dermal papillae, and also in hair follicles from other body sites, in order to elucidate the mechanism of androgen action on the hair growth process and related diseases

Changes of 5 alpha-reductase type II activity in sexual gland of diabetic male rats / 中华男科学杂志

<p><b>OBJECTIVES</b>To study the changes of sexual gland 5 alpha-reductase type II activity in pubertal and adult rats with diabetes.</p><p><b>METHODS</b>We selected 40 and 90 days old male Wistar rats as pubertal and adult animal model respectively, 30 rats in each group. The rats were randomly divided into three groups: control group (C), diabetic group (D) and diabetes with insulin replacement group (ID). The activity of 5 alpha-reductase type II was measured with thin layer chromatography in the epididymis, prostate and testis.</p><p><b>RESULTS</b>1. In all sexual glands of pubertal rats, the activity of 5 alpha-reductase type II in D group is significantly lower than that in C and ID groups. 2. In all sexual glands of adult rats. there is no difference in the activity of 5 alpha-reductase type II among these groups.</p><p><b>CONCLUSIONS</b>The activity of 5 alpha-reductase type II is likely to be influenced by metabolic environment, hormonal levels and local specific factors in pubertal rats, but it is relatively stable in adult rats.</p>

novel mutation in the androgen receptor gene causing partial androgen insensitivity syndrome

In the present study, a novel mutation in exon 7 of the androgen receptor [AR] gene in an Egyptian patient with partial androgen insensitivity syndrome [PAIS] was described. A male patient aged seven months was presented with ambiguous genitalia; the parents were not consanguineous. The patient had 46, XY karyotype and normal testosterone levels. Both basal and after human chorionic gonadotrophin [HCG] testosterone/dihydrotestosterone ratio was within normal suggesting normal 5-alpha reductase activity. Sequencing analysis of the AR gene revealed a novel mutation [P817A] within the ligand-binding domain [LBD]

Normal bone density in male pseudohermaphroditism due to 5alpha- reductase 2 deficiency

Bone is an androgen-dependent tissue, but it is not clear whether the androgen action in bone depends on testosterone or on dihydrotestosterone. Patients with 5alpha-reductase 2 deficiency present normal levels of testosterone and low levels of dihydrotestosterone, providing an in vivo human model for the analysis of the effect of testosterone on bone. OBJECTIVE: To analyze bone mineral density in 4 adult patients with male pseudohermaphroditism due to 5alpha-reductase 2 deficiency. RESULTS: Three patients presented normal bone mineral density of the lumbar column (L1-L4) and femur neck, and the other patient presented a slight osteopenia in the lumbar column. CONCLUSION: Patients with dihydrotestosterone deficiency present normal bone mineral density, suggesting that dihydrotestosterone is not the main androgen acting in bone

Estudio in vitro de tres antiandrógenos empleados en la actualidad en enfermedades andrógenodependientes / In vitro assay of three antiandrogens presently used in the treatment of androgen dependent diseases

Objetivo: Analizar el efecto in vitro del acetato de ciproterona, finasteride y flutamida, sobre la enzima 5Ó-reductasa, principal indicador bioquímico responsable del potente efecto andrógenico de la tetosterona, al convertirla en dihidrotestosterona. Material y Métodos: Se midio la actividad de la enzima 5Ó-reductasa en la próstata de ratas de macho adultos, utilizando concentraciones de 20 a 500µM de cada antiandrógeno. Resultados: El análisis estadístico muestra que la flutamina presenta mejor actividad antiandrogénica a medida que se incrementa su concentración, mientras que en el acetato de ciproterona y el finasteride, el efecto antiandrogénico fue menor a diferentes concentraciones (p<0.05); probablemente por una mayor velocidad de disociación de estos compuestos con su receptor