Serum inhibin B levels in males with gonadal dysfunction

OBJECTIVE: To determine whether inhibin B levels are reflective of the etiology of gonadal dysfunction. DESIGN: Institutional study. SETTING: A tertiary care university-affiliated infertility clinic. PATIENT(S): Forty-four men: 16 with primary testicular failure, 10 with partial idiopathic hypogonadotropic hypogonadism (IHH), 8 with primary germ cell failure, one with iatrogenic hypogonadotropic hypogonadism, one with untreated Kallmann's syndrome, and 8 healthy fertile controls. INTERVENTION(S): Three individuals (one each with IHH, hypogonadotropic hypogonadism [HH], and Kallmann's syndrome) underwent treatment with human chorionic gonadotropin. MAIN OUTCOME MEASURE(S): Baseline serum inhibin B, FSH, LH, total testosterone and estradiol levels, and sperm concentrations were measured. RESULT(S): Serum inhibin B concentrations were significantly higher in fertile controls (255 +/- 59 pg/mL) than in men presenting with primary testicular failure (75 +/- 46 pg/mL, P<.0001) or in those presenting with primary germ cell failure (73 +/- 31 pg/mL, P<.0001). Inhibin B levels were also lower in males with partial IHH (187 +/- 112 pg/mL, P<.05). The patient with iatrogenic HH had a level of 184 pg/mL, whereas the patient with Kallmann's syndrome had nondetectable levels (<10 pg/mL). Serum inhibin B levels correlated positively with sperm concentration (P=.0001), and negatively with FSH levels (P=.01) and LH levels (P<.05). Human chorionic gonadotropin therapy altered inhibin B levels. CONCLUSION(S): Inhibin B plays an important role as an endocrine regulator of FSH secretion, whereas gonadotropins are involved in the regulation of inhibin B secretion.     

AZFa candidate gene deletions in Taiwanese patients with spermatogenic failure.

BACKGROUND AND PURPOSE: Deletions of the azoospermia factor subregion a (AZFa) genes in proximal Yq11 are not frequently reported. The majority of AZFa deletions are thought to be associated with more severe testicular phenotypes, such as Sertoli cell-only syndrome. There is a lack of data on AZFa gene deletions in East Asian populations. In this study, we investigated the deletion status of AZFa genes in Taiwanese men with spermatogenic failure. METHODS: One hundred and eighty-three consecutive men with severe oligozoospermia or non-obstructive azoospermia were enrolled in this study. Genomic DNA was extracted from peripheral blood samples and polymerase chain reaction (PCR) was performed using primers specific to four AZFa genes: AZFaT1, DFFRY, DBY, and UTY. Sequence-tagged site markers (sY740, sY630, sY86, sY85, sY87, sY709, and sY88) were used to define the position of deletions. One hundred and twenty fertile men with normal spermatogenesis were enrolled as controls. RESULTS: Of the 183 patients, two showed single AZFa gene deletions, resulting in an overall frequency of 1.1%. One of these two patients had DFFRY deletion and the other had DBY deletion; their testicular phenotypes were Sertoli cell-only syndrome and hypospermatogenesis, respectively. Neither patient had deletions extending from AZFa through AZFb or AZFc. CONCLUSION: Our results suggest that AZFa gene deletion is infrequent in Taiwanese patients with severe oligozoospermia or non-obstructive azoospermia.     

Pulsatile luteinizing hormone-releasing hormone treatment of male hypogonadotropic hypogonadism

Luteinizing hormone-releasing hormone (LH-RH) secretion from the hypothalamus follows a rhythmic pattern, inducing pulsatile luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion from the pituitary gland. Consideration of this physiologic principle led to the introduction of pulsatile LH-RH therapy via infusion pump for the treatment of different forms of hypogonadotropic hypogonadism. We report on 10 male patients, 16 to 28 years of age, suffering from idiopathic hypogonadotropic hypogonadism (IHH) including Kallman's syndrome (n = 2) and delayed puberty (n = 2). All presented with complete eunuchoidism and had undergone no treatment for their conditions during the previous 2 years. LH-RH was administered in subcutaneous pulses of 4 to 16 micrograms, with a portable infusion pump (ZYKLOMAT, Ferring Corp., Kiel, FRG); treatment periods ranged from 6 to 24 months. With therapy, the subjects improved secretion of LH, FSH and testosterone. Testicular volumes and penis size increased; all patients developed normal secondary sexual characteristics. Spermatogenesis was induced in all patients. The time to onset of spermatogenesis ranged from 3 to 15 months. No major side effects were observed, and no patient dropped out of the study. The results indicate that pulsatile LH-RH therapy is an highly effective treatment for IHH and delayed puberty.     

Kallmann's syndrome: pregnancy through intracytoplasmic sperm injection and complicated by gestational diabetes.

Kallmann's syndrome is characterized by hypogonadotropic hypogonadism and anosmia. Assisted reproductive techniques such as intracytoplasmic sperm injection (ICSI) may be required to treat the infertile couple with oligozoospermia. Rare complications have been described in patients with Kallmann's syndrome, but gestational diabetes has not previously been reported. A case of Kallmann's syndrome with infertility is reported. Ovulation was successfully induced by human menopausal gonadotropin therapy, but pregnancy could not be achieved by artificial insemination or by conventional in vitro fertilization, although the husband had only moderate oligozoospermia. A high fertilization rate of the retrieved oocytes and successful pregnancy was achieved by ICSI. The pregnancy was complicated by gestational diabetes that was managed by insulin therapy. Successful ovulation induction in Kallmann's syndrome is not rare, but ICSI may be needed in selected cases. Some recent data have suggested that diabetes may occur in patients with Kallmann's syndrome, but further investigation is needed to establish whether gestational diabetes is associated with Kallmann's syndrome or is purely coincidental.     

Successful induction of ovulation using highly purified follicle-stimulating hormone in a woman with Kallmann's syndrome

OBJECTIVE: To describe a woman with Kallmann's syndrome who was treated successfully with highly purified FSH to achieve ovulation induction and pregnancy. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 32-year-old woman with Kallmann's syndrome who had been treated with oral contraceptives to prime secondary sex characteristics and genital organs since the age of 16 years. INTERVENTION(S): Highly purified FSH was administered intramuscularly for a total dose of 3,825 IU. MAIN OUTCOME MEASURE(S): Follicle number and diameter. RESULT(S): Three follicles with a diameter of > 1.7 cm and an endometrial thickness of 8 mm were observed. A clinical pregnancy, which subsequently was spontaneously aborted, was obtained. CONCLUSION(S): In primed patients with Kallmann's syndrome, highly purified FSH may be a useful alternative to pulsatile GnRH or menopausal gonadotropins to achieve ovulation induction and pregnancy.     

Association of AccI polymorphism in the follicle-stimulating hormone beta gene with polycystic ovary syndrome

Objective: To search for FSH β-subunit gene mutations in patients with polycystic ovary syndrome (PCOS) and determine the association between the mutations and the syndrome.

Design: Clinical and molecular studies.

Setting: Clinics and laboratories of the National University Hospital Obstetrics and Gynecology Department in Singapore.

Patient(s): One hundred thirty-five patients with PCOS and 105 normal control subjects.

Intervention(s): Exons two and three were screened for mutations by single-stranded conformational polymorphism and DNA sequencing.

Main Outcome Measure(s): Polymerase chain reaction followed by restriction enzyme analysis.

Result(s): No missense mutation was found in the functional units of the FSHβ gene in patients with PCOS, but a thymine-cytosine substitution in exon 3 (codon 76, TAT to TAC) was identified. The nucleotide change led to creation of an AccI digestion site. The distribution pattern of AccI polymorphism in the patients was significantly different from that in the control group, and the occurrence of homozygous carriers was significantly higher in patients (12.6%) than in the control group (3.8%). The frequency of polymorphism and prevalence of homozygosity were significantly higher in patients with PCOS with obesity (0.50% and 31.0%, respectively) than in those with menstrual disorders only (0.366% and 8.5%, respectively), which correlated with significantly higher androgen levels in the obese patients.

Conclusion(s): The AccI polymorphism in FSHβ gene may be associated with PCOS in some women, especially those with obesity.     

Mutations in KISS1 are not responsible for idiopathic hypogonadotropic hypogonadism in Chinese patients

PurposeTo investigate whether mutations in the KISS1 gene are present in 170 Chinese patients with idiopathic hypogonadotropic hypogonadism (IHH).MethodsMutational screening of the KISS1 gene was performed in 170 Chinese patients with IHH (133 male cases and 37 female cases) and 187 matched controls (94 males and 93 females).ResultsTwo known single-nucleotide polymorphisms (SNP), c. 58G > A in exon 1 and c. 242C > G in exon 2, were identified. However, no difference of genotype and allelic frequencies between cases and controls was observed.ConclusionsThe results suggest that mutations in the coding sequence of KISS1 are not common in patients with IHH in this Chinese population.     

Genetics and male infertility

This article reviews chromosomal and genetic disorders in the context of male fertility. Particular emphasis is on those disorders, which are encountered, in clinical practice including Klinefelter's syndrome, Kallman's syndrome, Androgen insensitivity, Y microdeletions, Y fertility gene deletions, and cystic fibrosis gene mutations. These disorders are discussed in relation to the aetiology of male fertility and also risks to children who are born of fathers with these disorders. A list of fathers' categories is proposed for outcome studies for children born after IVF-ICSI. Finally a question is proposed to catalyse debate about germ line therapy.     

Pulsatile gonadotropin-releasing hormone therapy in male patients with Kallmann's syndrome or constitutional delay of puberty

The response to low-dose pulsatile gonadotropin-releasing hormone (GnRH) therapy was tested in three hypogonadotropic hypogonadal male patients and a boy with delayed puberty showing different luteinizing hormone responses (delta LH) to test doses of 25 micrograms GnRH intravenously before treatment. Four male patients, 16 to 20 years of age, three with Kallmann's syndrome and one with idiopathic delay of puberty, received 2 micrograms GnRH subcutaneously every 2 1/2 hours for 3 months by the use of the Zyklomat pump (Ferring GmbH, Kiel, FRG). In two patients with Kallmann's syndrome and decreased delta LH, serum testosterone did not increase during treatment, even after increasing the dosage and changing the route of administration (4 micrograms subcutaneously or 8 micrograms intravenously every 2 1/2 hours for 4 weeks with every dosage). The third patient with Kallmann's syndrome and the boy with delayed puberty, both with normal delta LH, presented normal serum testosterone after 3 months of subcutaneous low-dose treatment. The different responses to a GnRH test dose corresponding to the response to pulsatile GnRH therapy probably reflect different degrees of maturation of the pituitary gonadotrophs.     

The genetics of hypogonadotropic hypogonadism

An up-to-date review of the genetic aspects of idiopathic hypogonadotropic hypogonadism (IHH)/Kallmann syndrome (KS) is presented. Because proper development of the neuroendocrine axis must occur for normal puberty and reproductive function, gonadotropin-releasing hormone (GnRH) neuron migration is outlined first, followed by an introduction to the in vitro analysis of GnRH neuron migration. The normal hypothalamic-pituitary-gonadal (HPG) axis at different ages is discussed, along with a brief overview of normal and delayed puberty in both boys and girls. The phenotype of IHH/KS is discussed in detail, with its relation to Mendelian inheritance and chromosomal translocations. The molecular basis of IHH/KS is reviewed, with particular emphasis on the three most common genes ( KAL1, FGFR1, and GNRHR) that possess mutations in these patients. However, all other known genes for which mutations occur are also addressed briefly. The goal of this review is to provide a comprehensive discussion of IHH/KS, and to include both basic science and clinical findings that should allow a more complete understanding of hypothalamic-pituitary neuroendocrinology that is important in puberty and reproduction.     

RAS/RAF mutation and defective DNA mismatch repair in endometrial cancers

OBJECTIVE: Defective DNA mismatch repair is a common genetic abnormality in both colon cancers and endometrial cancers. Cancers with defective DNA mismatch repair have the so-called mutator phenotype and accumulate genetic errors at an increased rate. An early mutational target in cells with defect DNA mismatch repair may be the RAS/RAF pathway. Colon cancers often have KRAS2 mutations and, if not KRAS2 mutations, may have BRAF mutations. This study investigated the spectrum and frequency of mutations in BRAF and KRAS2 in endometrial carcinomas on the basis of mismatch repair status. STUDY DESIGN: Four hundred forty-one patients with endometrial cancer were staged properly and graded and evaluated for mismatch repair status. These patients were then stratified to groups by the degree of microsatellite instability that was observed in their tumors. One hundred forty-six of the selected tumors were then evaluated for KRAS2 and BRAF mutations on the basis of their microsatellite instability. RESULTS: One hundred forty-six endometrioid endometrial cancers were evaluated for KRAS2 and BRAF mutations. Thirty-five cancers (24%) had activating KRAS2 mutations, but only a single BRAF mutation was identified in an microsatellite instability-positive cancer. Twenty-four of 81 microsatellite instability high cancers (29.6%) in which the MLH1 repair gene was methylated had KRAS2 mutations. When compared with the other groups, this finding approached statistical significance (P=.06). KRAS2 mutation status was associated with increasing age at diagnosis (P=.02). CONCLUSION: Despite many similarities between colon and endometrial cancers, the mechanism of the development of endometrial cancers appears to be different from colon cancers in that BRAF is not affected by a mismatch repair problem, because only KRAS2 mutations were seen. In addition, increasing age appears to lead to an increased likelihood that such a mutation will occur.     

Gonadotropin-releasing hormone, follicle-stimulating hormone beta, luteinizing hormone beta gene structure in idiopathic hypogonadotropic hypogonadism.

OBJECTIVE: To determine if the genes for gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone beta (FSH beta), and luteinizing hormone beta (LH beta) are present, and if so, whether gene structure is normal in patients with idiopathic hypogonadotropic hypogonadism (IHH). DESIGN: Patients with clinical and laboratory characteristics of IHH were studied at the deoxyribonucleic acid (DNA) level to assess gene structure. SETTING: This study took place in an academic setting. PATIENTS: Human volunteers with documented IHH and fertile controls were studied. INTERVENTIONS: Genomic DNAs were extracted from each patient, Southern blots were constructed and hybridized to DNA probes for GnRH, FSH beta, and LH beta. DNA samples were also subjected to polymerase chain reaction analysis. MAIN OUTCOME MEASURES: Gene structure was assessed by analysis of autoradiographs and gel electrophoresis of polymerase chain reaction products in both the study patients and controls. RESULTS: Each analysis for FSH beta, LH beta, and GnRH demonstrated the same sized fragments in both the study group and control group. A 1.2-kilobase fragment containing the coding region for GnRH was present in all patients with IHH and controls by polymerase chain reaction. CONCLUSIONS: The genes for GnRH, LH beta, and FSH beta are present in patients with IHH. No large deletions or rearrangements of any of these genes were identified in any of these patients.     

Molecular epidemiologic study of mitochondrial DNA mutations in patients with mitochondrial diseases in Taiwan.

We report an 8-year molecular study of mitochondrial DNA (mtDNA) mutations in patients with mitochondrial diseases in Taiwan. One hundred and seventy-seven patients met the diagnostic criteria of mitochondrial disease and were recruited into the study. The results showed that 32 patients, including 25 with mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome, one with Kearns-Sayre syndrome (KSS), one with diabetes mellitus and deafness, and five with chronic progressive external ophthalmoplegia (CPEO), harbored the A3243G mtDNA mutation. The A8344G mutation was found in nine patients, all of whom suffered from myoclonic epilepsy and ragged-red fibers (MERRF) syndrome. The G11778A mtDNA mutation was found in 18 of 22 patients with Leber's hereditary optic neuropathy. The T8993C and T8993G mutations were found, respectively, in one and two patients with Leigh syndrome. Large-scale deletions of mtDNA were found in 17 patients with CPEO, one with KSS, one with MELAS, and two with MERRF syndrome. The mtDNA mutations in patients with each of the mitochondrial diseases found in Taiwan were restricted mainly to a single site, while those reported for the same diseases in other ethnic groups occurred in many sites. Furthermore, significant levels of additional mtDNA mutations occurred in some patients with mitochondrial encephalomyopathies. We suggest that these additional (or secondary) mtDNA mutations are generated as a consequence of the preexisting primary mtDNA mutations and may contribute to the age-dependent progressive deterioration characteristic of mitochondrial diseases.     

重度子痫前期患者及其新生儿线粒体三功能蛋白酶α亚单位G1528C基因突变的研究

目的 探讨线粒体三功能蛋白酶（ MTP）α亚单位G1528C基因突变,在重度子痫前期及溶血、肝酶升高和低血小板计数（HELLP综合征）孕妇及其新生儿中的携带情况.方法 北京地区汉族重度子痫前期（子痫前期组）孕妇及其新生儿130例（其中合并肝损害18例）,HELLP综合征（HELLP组）孕妇及其新生儿10例,同期无妊娠期特发性肝损害等并发症及其他代谢性疾病的正常孕妇90例及新生儿560例作为对照组.应用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）技术分析3组孕妇外周血及其新生儿脐血中MTPα亚单位G1528C基因突变的携带情况.实验中阳性对照标准为西方白种人的G1528C杂合子标本.结果 3组孕妇外周血及其新生儿脐血中的MTPα亚单位G1528C基因电泳图中均无与阳性对照标本中相应分子质量的条带出现.结论 MTP的α亚单位G1528C基因突变可能不是北京汉族人种常见的突变位点; MTP缺陷在北京汉族人与西方白种人之间可能存在着种族差异.     

年轻子宫内膜癌患者保留生育能力治疗的思考

最新发布的《CA CANCER J CLIN》报道称,2019年美国有70余万例子宫内膜癌患者,有超过5万的新发病例。子宫内膜癌已成为美国发病率排名第二的妇科肿瘤,仅次于乳腺癌［1］。随着经济及生活水平的提高,以及生育年龄延迟,我国子宫内膜癌的发病率越来越接近发达国家。据最新一期国家癌症中心数据,我国2015年子宫内膜癌新发病例居我国恶性肿瘤发病前10位,占女性全部恶性肿瘤发病的3.88%,这一数据也较2014年的3.79%有所上升。城市地区的子宫内膜癌发病率为11.35/10万,高于农村地区;在上海,子宫内膜癌新发病例已超过子宫颈癌［2］。 浏览更多请关注本刊微信公众号及当期杂志。     

A predictive index of cure versus no cure in advanced ovarian carcinoma patients—Replacement of second-look laparotomy as a diagnostic test

Three hundred forty-two Stage III and IV epithelial ovarian carcinoma patients received cytoreductive surgery followed by Adriamycin and cisplatin, 50 mg/m2 each, q 4 weeks for 9 courses. One hundred ninety-seven were clinically NED at completion of treatment and 173 of these 197 had a second-look laparotomy. One hundred twenty had persistent disease. Fifty-three were second-look negative and had no further treatment. Thirty of these latter patients relapsed--all (with one exception) within 2 years. Those not relapsing after negative second-look are considered "cured" (median follow-up 42 months, range 24-68 months) and all others "failures." Stage was a significant predictor of treatment failure--there were no Stage IV "cures." In Stage III patients, age and largest residual tumor diameter post initial surgery were significant predictors of failure. Performance status was marginally significant. In our series, any patient with Stage IV disease or Stage III disease with at least two of the following three poor prognostic factors had a chance of cure of 2.2% (2 "cures" out of 90 patients): age greater than 60 years, macroscopic residual initially, or initial performance status of 2 or 3. Under normal circumstances a second-look procedure to identify persistent disease in this group of patients does not appear justified.     

亚甲基四氢叶酸还原酶、凝血因子V和凝血酶原基因多态性与原因不明复发性早期流产的关系

目的探讨亚甲基四氢叶酸还原酶（MTHFR）基因C677T突变、凝血因子V（FV）基因G1691A突变和凝血酶原（PT）基因G20210A突变与原因不明复发性早期流产（URESA）的关系。方法应用聚合酶链反应．限制性片段长度多态性（PCR-RFLP）技术分析112例URESA患者（病例组）和100例健康妇女（对照组）MTHFR、FV和PT3种基因多态性。结果（1）MTHFR基因的T／T基因型和T等位基因频率,病例组[分别为38．4％（43／112）和59．8％（134／224）]高于对照组[分别为18．0％（18／100）和43．0％（86／200）],两组分别比较,差异均有统计学意义（P＜0．01）。病例组与对照组比较,T／T基因型者发生URESA的相对风险增加（OR=2．8390,95％CI为1．5022～5．3661）。（2）病例组和对照组妇女Fv基因和PT基因均为G／G基因型（即正常带型）。结论MTHFR基因C677T位点多态性与URESA发病密切相关,T／T基因型是其发病的危险因素。FV和PT基因的突变率在中国妇女人群中极低。     

Metformin reduces abortion in pregnant women with polycystic ovary syndrome.

BACKGROUND: Women with polycystic ovary syndrome (PCOS) are considered to be at increased risk of miscarriage. Since metformin has beneficial effects on the risk factors contributing to first-trimester abortion in PCOS patients, we hypothesized that metformin - owing to its metabolic, endocrine, vascular and anti-inflammatory effects - may reduce the incidence of first-trimester abortion in PCOS women. MATERIALS AND METHODS: A prospective cohort study was set up to determine the beneficial effects of metformin on PCOS patients during pregnancy. Two hundred non-diabetic PCOS patients were evaluated while undergoing assisted reproduction. One hundred and twenty patients became pregnant while taking metformin, and continued taking metformin at a dose of 1000-2000 mg daily throughout pregnancy. Eighty women who discontinued metformin use at the time of conception or during pregnancy comprised the control group. RESULTS: Both groups were similar with respect to all background characteristics (age, body mass index, waist/hip ratio, follicle-stimulating hormone, luteinizing hormone, estradiol and dehydroepiandrosterone sulfate levels). Rates of early pregnancy loss in the metformin group were 11.6% compared with 36.3% in the control group (p < 0.0001; odds ratio = 0.23, 95% confidence interval 0.11-0.42). CONCLUSIONS: Administration of metformin throughout pregnancy to women with PCOS was associated with a marked and significant reduction in the rate of early pregnancy loss.     

Molecular analysis of defects in the CFTR gene and AZF locus of the Y chromosome in male infertility

OBJECTIVE: To investigate the frequency and potential impact of mutations and polymorphisms in the CFTR gene and deletions in AZF locus of the Y chromosome in patients with azoospermia (AZOO), cryptozoospermia (CRYPTO) or oligoasthenoteratozoospermia (OAT) who were to be included in an assisted reproductive technologies (ART) program. STUDY DESIGN: A total of 188 infertile men were enrolled in the study: 100 patients with AZOO, 38 with CRYPTO and 50 with OAT. RESULTS: The CFTR gene mutations or IVS8-5T variant in at least 1 allele was identified with similar frequencies among the AZOO (33%) and CRYPTO (21%) patients; 55% of the AZOO patients with normal spermatogenesis (NS) had mutations in 1 or 2 alleles. The novel R810G mutation in exon 13 was identified in 1 NS patient. The OAT or AZOO patients with Sertoli cell only syndrome (SCO) had mutations in the CFTR gene with similar frequencies to that in the general Polish population. The deletions in the AZF locus were detected in 20% of SCO patients, 11.5% of AZOO patients with maturation arrest and in 5% of CRYPTO patients. The other groups (NS, OAT) did not carry deletions in the region studied. CONCLUSION: Molecular diagnosis of the CFTR gene, Y chromosome deletion analysis and genetic counseling are necessary diagnostic elements for patients with male infertility, especially if the are included in an ART program.     

Screening for mutations in the SRY gene in patients with mixed gonadal dysgenesis or with Turner syndrome and Y mosaicism

OBJECTIVE: To investigate the presence of mutations in the open reading frame (ORF), as well as on the 5' and 3', flanking regions of the SRY gene in patients with mixed gonadal dysgenesis (MGD) or with Turner syndrome (TS) and Y mosaicism. STUDY DESIGN: We studied 13 patients with MGD and three patients with TS and Y mosaicism. DNA was isolated from blood leukocytes for subsequent polymerase chain reaction (PCR) and direct sequencing were performed in the ORF, as well as from the 5' and 3' flanking regions of the SRY gene. RESULTS: No mutations were present in any of the patients studied. CONCLUSION: The absence of mutations in these regions indicated that mutations were an unlikely cause of MGD or TS with Y mosaicism and suggested that there are others genes playing an important role in sex development.