Idiopathic Premature Ovarian Failure Research Articles

Diagnosis of Idiopathic Premature Ovarian Failure by Color Doppler Ultrasound under the Intelligent Segmentation Algorithm.

The aim of this study was to explore the application value of transvaginal color Doppler ultrasound based on the improved mean shift algorithm in the diagnosis of idiopathic premature ovarian failure (POF). In this study, 80 patients with idiopathic POF were selected and included in the experimental group, and 40 volunteers who underwent health examinations during the same period were selected and included in the control group, who underwent transvaginal Doppler ultrasound examination. At the same time, an improved mean shift algorithm was proposed based on artificial intelligence technology and applied to ultrasound image processing. In addition, the ovarian artery parameters of patients were compared in two groups, including peak systolic flow rate (PSV), diastolic flow rate (EDV), resistance index (RI), and pulsatile index (PI). The results showed that the relative difference degree (RDD) of the segmentation results of the algorithm in this study was significantly lower than that of Snake, Live_wire, and the traditional mean shift algorithm, while the relative overlap degree (ROD) and Dice coefficient were opposite, and the differences were significant (P<0.05). The mediolateral diameter of control group was 2.87±0.31cm, and the anteroposterior diameter was 1.86±0.28 cm; while those were 2.11±0.36 cm and 1.13±0.34 cm, respectively, in the experimental group, showing significant differences between the groups (P<0.05). Of the 80 patients in the experimental group, 132 cases with ovarian arteries were found; among 40 patients in the experimental group, 76 cases were found with ovarian arteries, and the hemodynamic detection rate of the experimental group was significantly lower than that of the control group (P<0.05). The ovarian artery parameters PI, RI, and S/D of the experimental group were significantly higher than those of the control group, and the differences were statistically significant (P<0.05). The results showed that the segmentation results of the improved algorithm in this study were more superior to the segmentation results of other algorithms. The regional information loss of the segmentation results was not serious, and the resolution was higher and the definition was higher. The transvaginal color Doppler ultrasound based on the artificial intelligence segmentation algorithm can clearly show the functional status and hemodynamics of the patient's ovaries. The ovarian artery parameters PI and RI can be used as specific indicators for evaluating the POF.

P–541 Identification of novel variants and candidate genes in women with familial idiopathic premature ovarian failure using whole-exome sequencing

Abstract Study question Is it possible to identify a genetic cause of familial premature ovarian failure (POF) with whole-exome sequencing (WES)? Summary answer Whole-exome sequencing is the most efficient strategy to identify probably pathogenic mutations in different genes in pathologies of polygenic etiology such as premature ovarian failure. What is known already Premature ovarian failure is the loss of ovarian function before the age of 40, and it is a common cause of infertility in women. This pathology has a heterogeneous etiology. Some chromosomal and genetic alterations have been described, and could explain approximately 20% of cases. However, in most patients the origin remains unknown. Recent studies with next-generation sequencing (NGS) have identified new variants in candidate genes related with premature ovarian insufficiency (POI) or premature ovarian failure (POF). These genes are not only involved in processes such as folliculogenesis, but also with DNA damage repair, homologous recombination, and meiosis. Study design, size, duration Fourteen women, from 7 families, affected by idiopathic POF were included in the study from October 2019 to September 2020. Seven POF patients were recruited when they came to our clinic to undergo assisted reproductive treatment. In the anamnesis, it was found that they had relatives with a diagnosis of POF, who were also recruited for the study. The inclusion criteria were amenorrhea before 38 years old and analytical and ultrasound signs of ovarian failure. Participants/materials, setting, methods WES was performed using TrusightOne (Illumina®). Sequenced data were aligned through BWA tool and GATK algorithm was used for SNVs/InDel identification. VCF files were annotated using Variant Interpreter software. Only the variants shared by each family were extracted for analysis and these criteria were followed: (1) Exonic/splicing variants in genes related with POF or involved in biological ovarian functions (2) Variants with minor allele frequency (MAF) ≤0.05 and (3) having potentially moderate/strong functional effects. Main results and the role of chance Seventy-nine variants possibly related with the POF phenotype were identified in the seven families. All these variants had a minor allele frequency (MAF) ≤0.05 in the gnomAD database and 1000 genomes project. Among these candidate variants, two were nonsense, six splice region, one frameshift, two inframe deletion and 68 missense. Thirty-two of the missense variants were predicted to have deleterious effects by minimum two of the four in silico algorithms used (SIFT, PolyPhen–2, MutationTaster and PROVEAN). All variants were heterozygous, and all the families carried three or more candidate variants. Altogether, 43 probably damaging genetic variants were identified in 39 genes expressed in the ovary and related with POF/POI or linked to ovarian physiology. We have described genes that have never been associated to POF pathology, however they may be involved in key biological processes for ovarian function. Moreover, some of these genes were found in two families, for example DDX11, VWF, PIWIL3 and HSD3B1. DDX11 may function at the interface of replication-coupled DNA repair and sister chromatid cohesion. VWF gene is suggested to be associated with follicular atresia in previous studies. PIWIL3 functions in development and maintenance of germline stem cells, and HSD3B1 is implicated in ovarian steroidogenesis. Limitations, reasons for caution Whole-exome sequencing has some limitations: does not cover noncoding regions of the genome, it also cannot detect large rearrangements, copy-number variants (large deletions/duplications), mosaic mutations, mutations in repetitive or high GC rich regions and mutations in genes with corresponding pseudogenes or other highly homologous sequences. Wider implications of the findings: WES has previously shown to be an efficient tool to identify genes as cause of POF, and has demonstrated the polygenic etiology. Although some studies have focused on it, and many genes are identified, this study proposes new candidate genes and variants, having potentially moderate/strong functional effects, associated with POF. Trial registration number Not applicable

Evaluation of safety, feasibility and efficacy of intra-ovarian transplantation of autologous adipose derived mesenchymal stromal cells in idiopathic premature ovarian failure patients: non-randomized clinical trial, phase I, first in human

BackgroundPremature ovarian failure (POF) is characterized by the loss of ovarian activity before the age of 40 years. Stem cell therapy has the capability to create a regenerative microenvironment and is a proposed treatment for POF-related infertility due to the presence of renewal folliculogenesis and germ cells in the adult ovaries. In this study, we assessed the safety, feasibility, efficacy and dose adjustment of autologous adipose-derived stromal cells (ADSCs) and their ability to improve ovarian function in POF patients.MethodsThis study was a non-randomized clinical trial, phase I. Nine women with a definitive diagnosis of POF were divided into three groups (n = 3 per group) that received either 5 × 106, 10 × 106, or 15 × 106 autologous ADSCs suspension transplanted in the one ovary. Participants were followed-up at 24 h after the transplantation, and at 1 and 2 weeks, and 1, 2, 3, 6, and 12 months after the transplantation. The primary objective was to evaluate the safety of ADSCs transplantation. Secondary objectives included the effects of ADSCs transplantation on the resumption of menstruation, hormones level (Follicle-stimulating hormone (FSH) and anti-Müllerian hormone), ovarian function (Antral follicle count and ovary volume by ultrasonography evaluation) as well as dose escalation.ResultsParticipants had not shown any early-onset possible side effects and secondary complications during follow-up. The menstruation resumption was observed in four patients which established for several months. In the 15 × 106 group, two POF patients had a return of menstruation second months after the intervention. Two other POF patients in 5 × 106 and 10 × 106 cell groups reported menstruation resumption at 1 month after the intervention. We observed decreased serum FSH levels of less than 25 IU/l in four patients. In two patients in 5 × 106 and 10 × 106 cell groups, serum FSH showed an inconsistent decline during a 1 year follow up after ADSCs transplantation. The ovarian volume, AMH, and AFC were variable during the follow-up and no significant differences between cell groups (p > 0.05).ConclusionsWe showed the intra-ovarian embedding of ADSCs is safe and feasible and is associated with an inconsistent decline in serum FSH. This should be further investigated with a large RCT.Trial registrationNCT02603744, Registered 13 November 2015 - Retrospectively registered, http://www.Clinicaltrials.gov

The Predictive Value of Osteocalcin, Granulin, Cathepsin K and Some Other Biomarkers in Women with Premature Ovarian Failure

Background: Premature ovarian failure (POF), often and misleadingly referred to as ‘premature menopause’,is defined as a loss of ovarian activity before the age of 40 years and is characterized by irregular or absentperiods and reduced fertility. Symptoms include those associated with the natural menopause (night sweatsand vaginal dryness), and with the long-term adverse effects of estrogen deficiency (osteoporosis andcardiovascular disease): the latter is believed to explain the shorter life expectancy associated with POF.Aim: To determinethe predictive value of serum osteocalcin(OC), granulin(GRN), cathepsin k (CTK),vitamin D, parathyroid hormones (PTH), calcium and phosphrus as biomarkers for POF.Method: Sixty (60) women with idiopathic POF with thirty (30) women as control groups were included inthe study. Baseline investigation in all subjects included serum Osteocalcin(OC), granulin(GRN), cathepsink (CTK), vitamin D, parathyroid hormones (PTH), calcium and phosphrus levels were estimated using theappropriate assays for each.Results: The mean serum levels of Osteocalcin, Ganulin,Parathyroid hormones,vitamin D and phosphorus,were significantly higher in women with POF,when compared to healthy controls (P=&lt;0.05). However, meanserum levels of cathepsin k and calcium in POF group were non significant differences,when compared tohealthy controls.Conclusion: Osteocalcin and granulin serum levels may be used as new biomarker for the diagnosis ofpremature ovarian failure.

Single Strand Conformation Polymorphism and Sequencing of HS6ST2 Gene in Patients of Idiopathic Premature Ovarian Failure

Single Strand Conformation Polymorphism and Sequencing of HS6ST2 Gene in Patients of Idiopathic Premature Ovarian Failure

Idiopathic Premature Ovarian Failure and its association to the abnormal longitudinal changes of telomere length in a population of Iranian Infertile Women: A pilot study

BachgroundThe premature ovarian failure (POF) is a reason of infertility that affects about 1–4% of women before age 40. The importance of telomeres' length in different diseases has been explored before. This study examines the association between the relative telomere length and idiopathic POF in a group of Iranian women. MethodsThe blood genomic DNA was extracted from 40 idiopathic POF patients (case group) and 40 fertile women (control group). The relative telomere length (RTL) was evaluated by quantitative Real-Time PCR using specific telomeric primers. RTL was calculated as T (telomere)/S (single copy gene) ratio and compared between infertile and fertile groups. ResultsA strong association was considered between telomere size and idiopathic premature ovarian failure. In patients the relative telomere length showed to be significantly longer than those of control group (P < .05, 95% CI). ConclusionOur findings demonstrate a possible relationship between telomere lengthening and idiopathic POF. The reason of the observed elevated telomeres' genetic material could be explained by numerous probable mechanisms like reduced ovarian cell division rate, a sudden increase in the estrogen level before menopause and after egg depletion, and/or an autoimmune condition which could change the composition of blood cells and their consequent conversion to the cells with longer telomeres in patients. More experiments with larger population are necessary to confirm the results of present study.

Premutations of FMR1 CGG repeats are not related to idiopathic premature ovarian failure in Iranian patients: A case control study

Premature ovarian failure (POF) is a reproductive disease which affects 1 in 100 under 40 years women. FMR1 premutation carriers of CGG repeats are supposed to be at increased risk for POF. We have examined the 5′UTR region of the gene to find any association between the repeat size and the disease etiology in Iranian population. 30 women with early idiopathic POF and 30 fertile control women were selected. We used triplet repeat primed PCR (TP PCR) assay and gene-specific primers to amplify the CpG Island of FMR1 gene promoter region. The amplification results were analyzed by capillary electrophoresis and Gene Marker software. Among 30 patients, two had intermediate repeat size, one had premutation and the rest had CGG repeat of the normal range. Two of controls had intermediate repeats and none had a premutation. Two groups had significant differences in the repeat number average (p = 0.007) and in the average length of the smallest allele (p < 0.001), but had no promising difference in average length of the longest allele (p = 0.453).Although the two groups showed a significant difference in the length of alleles, their length was within normal range. Perhaps the polymorphism, in connection with the genome's entire network, has been involved in the development of the disease, or there has been a fundamentally different mechanism for the disease in Iranian population. A larger number of Iranian POF patients should be investigated for any probable relationship between the CGG triplet repeat length and the etiology of the disease.

Inhibin α gene –16C → T and 769 G → A polymorphisms in Syrian women with idiopathic premature ovarian failure

Inhibin α gene –16C → T and 769 G → A polymorphisms in Syrian women with idiopathic premature ovarian failure

Bee Stings at Sites of Acupuncture as a Potential Therapy for Idiopathic Premature Ovarian Failure: A Pilot Study

Objectives:To evaluate administration of bee stings at sites of acupuncture on the hormonal profile of patients with idiopathic premature ovarian failure (POF). Study design: A pilot study carried at Obstetrics and Gynaecology Department, Tanta University Hospital, and at the Entomology Unit, Zoology Department, Faculty of Science, Tanta University from June 2015 to December 2015. Patients and Methods: 20 patients aged 18 to 40 years with idiopathic POF were included in a case series study. Measurement of serum concentrations of Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH), 17β-estradiol (E2) and Anti-Mullerian hormone (AMH) with calculation of Antral Follicles Count (AFC) and volume of ovaries was done before and after bee stings that were administered for 3 months, 2 sessions per week. Every session was 15 minutes with a range of 3-8 stings per session at the site of acupuncture. The outcome measures were changes in estimated variables within one month after bee stings. Results:FHS and LH significantly decreased from 54.5 ± 3.2 to 27.1 ± 5 mIU/mL and from 53.8 ± 2.7 to 25.2 ± 4.1 mIU/mL respectively (P value< 0.001). E2 significantly increased from 38.7 ± 3.3 to 92.5 ± 2.9 Pg/mL (P value< 0.05). Ovarian volume, AFC and AMH were not specifically different after bee stings.Conclusion:Bee stings at the sites of acupunctures significantly reduced the serum levels of both FSH and LH and increased serum concentrations of E2 with improvement in hot flushes and vaginal dryness. Bee stings had no effect on AFC, AMH or ovarian volume. The use of bee venom (stings) therapy at the sites of acupunctures for patients with idiopathic POF is recommended.

Premature Ovarian Failure: An Association with Autoimmune Diseases.

Premature Ovarian Failure (POF) is the cessation of ovarian function before the age of 40 years. POF is reported to be associated with autoimmune diseases in 20-30% of cases. Patients presenting with idiopathic POF were screened for the presence of autoimmune disorders. Twenty patients with idiopathic POF were included in the study. Baseline investigation in all subjects included fasting serum FSH, LH, E2, progesterone, free T3, free T4, Thyroid-Stimulating Hormone (TSH) and Anti-Thyroperoxidase (anti-TPO) antibodies, testosterone and Dehydroepiandrosterone (DHEAS) levels. Fasting and post-glucose (2 hours after 75g of oral glucose) serum calcium and phosphate were estimated using appropriate assays in biochemistry laboratory. Seven patients (35%), who presented with secondary amenorrhea, had thyroid disorders and were already on thyroxine replacement therapy. One patient also had vitiligo. There was no history of adrenal disorder. Anti-TPO levels were elevated in two (10%) patients of secondary amenorrhea group. The levels of serum testosterone were low in three patients. Serum DHEAS levels were low in 13 patients. Blood sugar levels (fasting and 2 hour post 75g glucose load) and fasting insulin levels were normal. Serum calcium and phosphate levels were normal in all the patients. Thyroid autoimmunity is the most common autoimmune disease associated with POF. The finding of low DHEAS in a large percentage of patients (65%), suggests possibility of adrenal dysfunction. This requires further testing for adrenal reserve and adrenal autoantibodies.

Analysis of FMR1 gene premutation and X chromosome cytogenetic abnormalities in 100 Tunisian patients presenting premature ovarian failure

ObjectiveTo evaluate the prevalence of FMR1 premutations and X chromosome cytogenetic abnormalities in a large cohort of Tunisian women with premature ovarian failure (POF). Patients and methodsThe cohort consisted of 127 Tunisian women with POF referred by endocrinologists and gynecologists for genetic investigation in the context of idiopathic POF and altered hormonal profiles. Clinical information concerning the reproductive function in the family, previous hormonal measurements and/or possible fertility treatment were collected. Karyotype, FISH analyses, FMR1 and FMR2 testing were performed for all patients. ResultsFifteen patients (11.81%) presented structural or numerical X chromosomal abnormalities. Moreover, we detected in 12 patients (10.71%) a high level of X mosaicism. Analysis of FMR1 gene in the 100 patients without X chromosomal abnormalities showed that five percent of the patients carried a FMR1 premutation allele. On the other hand, the FMR2 screening did not reveal any deletion. ConclusionOur study confirms the major role of X chromosome abnormalities in POF and highlights the importance of karyotype analyses and FMR1 screening. These investigations provide valuable information for diagnosis and genetic counseling for these women who still have a 5% chance of spontaneous conception.

The genetics of premature ovarian failure: current perspectives.

Premature ovarian failure (POF) is a common cause of infertility in women, characterized by amenorrhea, hypoestrogenism, and elevated gonadotropin levels in women under the age of 40. Many genes have been identified over the past few years that contribute to the development of POF. However, few genes have been identified that can explain a substantial proportion of cases of POF. The unbiased approaches of genome-wide association studies and next-generation sequencing technologies have identified several novel genes implicated in POF. As only a small proportion of genes influencing idiopathic POF have been identified thus far, it remains to be determined how many genes and molecular pathways may influence idiopathic POF development. However, owing to POF’s diverse etiology and genetic heterogeneity, we expect to see the contribution of several new and novel molecular pathways that will greatly enhance our understanding of the regulation of ovarian function. Future genetic studies in large cohorts of well-defined, unrelated, idiopathic POF patients will provide a great opportunity to identify the missing heritability of idiopathic POF. The identification of several causative genes may allow for early detection and would provide better opportunity for early intervention, and furthermore, the identification of specific gene defects will help direct potential targets for future treatment.

Human bone marrow mesenchymal stem cells therapy restores ovarian functions and reverses infertility in a preclinical model of chemotherapy induced premature ovarian failure

To determine the efficacy of xenogeneic human bone marrow mesenchymal stem cells (HBMSCs) in restorative functioning of chemotherapy-damaged ovaries in mice which will serve as a prelude to assess the utility of preserving fertility with adult stem cells among cancer survivors & for treatment of patients with idiopathic premature ovarian failure.

Granulin levels in patients with idiopathic premature ovarian failure

ObjectiveWith the prolongation of expected lifespan, premature ovarian failure (POF) has gained increased significance as a crucial female health problem. Here we aimed to investigate the diagnostic value of the levels of the growth factor Granulin, which has been associated with epithelial ovarian carcinoma, in the pathogenesis of POF. Study designThirty-one patients with POF and 57 patients as a control group were recruited for this study in a tertiary referral center in the capital of Turkey. Granulin levels and individual characteristics of the two groups were compared. ResultsAge and body mass indices were similar between the two groups. Granulin levels in patients with POF (2.94±1.91ng/mL) were significantly lower than in the control group (4.77±1.62ng/mL) (p<0.001). ConclusionFuture larger studies should focus on Granulin levels and its correlation with ovarian reserve markers, its clinical applicability, and its possible therapeutic value, thus providing further insight into the role of Granulin in the pathogenesis of POF.

Increased Incidence of Mitochondrial Cytochrome C Oxidase 1 Gene Mutations in Patients with Primary Ovarian Insufficiency.

Primary ovarian insufficiency (POI), also known as premature ovarian failure (POF), is defined as more than six months of cessation of menses before the age of 40 years, with two serum follicle stimulating hormone (FSH) levels (at least 1 month apart) falling in the menopause range. The cause of POI remains undetermined in the majority of cases, although some studies have reported increased levels of reactive oxygen species (ROS) in idiopathic POF. The role of mitochondrial DNA in the pathogenesis of POI has not been studied extensively. This aim of this study was to uncover underlying mitochondrial genetic defects in patients with POI. The entire region of the mitochondrial genome was amplified in subjects with idiopathic POI (n=63) and age-matched healthy female controls (n=63) using nine pair sets of primers, followed by screening of the mitochondrial genome using an Illumina MiSeq. We identified a total of 96 non-synonymous mitochondrial variations in POI patients and 93 non-synonymous variations in control subjects. Of these, 21 (9 in POI and 12 in control) non-synonymous variations had not been reported previously. Eight mitochondrial cytochrome coxidase 1 (MT-CO1) missense variants were identified in POI patients, whereas only four missense mutations were observed in controls. A high incidence of MT-CO1 missense variants were identified in POI patients compared with controls, and the difference between the groups was statistically significant (13/63 vs. 5/63, p=0.042). Our results show that patients with primary ovarian insufficiency exhibit an increased incidence of mitochondrial cytochrome c oxidase 1 gene mutations, suggesting that MT-CO1 gene mutation may be causal in POI.

Mutational analysis of the FIGLA gene in women with idiopathic premature ovarian failure.

The aim of our study was to identify the association of FIGLA (factor in the germ line α) gene variants with premature ovarian failure (POF) in the Indian population. Two hundred nineteen women with idiopathic POF and 230 healthy controls were recruited for this study. All exons, intron-exon boundaries, and untranslated regions of the FIGLA gene were analyzed by polymerase chain reaction and sequencing. All FIGLA variants were analyzed in silico, using PolyPhen, SIFT, MutationTaster, PMUT, I-Mutant, Mupro, Align-GVGD, PROVEAN, and HOPE software, to predict pathogenicity and changes in protein stability. Seven different FIGLA variants were detected among women with POF. Two exon 3 variants, c.427G → C and c.557C → T, showed strong association between cases and controls (P = 0.02 and P = 0.02, respectively). Significant differences in both of these variants were observed between cases and controls in genotype and dominant models. No significant difference between controls and women with POF was found on haplotype analysis. A nonsynonymous variant, p.(Arg83Cys), was found only in POF cases. Variant p.(Arg83Cys) lies in the functional domain of the FIGLA gene (basic helix-loop-helix), and protein alignments reveal that it is highly conserved among mammals. In silico analysis suggests the functional involvement of p.(Arg83Cys) and p.(Ser141Thr) variants in POF pathogenesis. We have established a strong association between FIGLA gene variants and POF in Indian women, which may be a potential genetic risk factor in the development of idiopathic POF. However, further independent genetic and functional studies are necessary to confirm our findings.

Association of FOXE1 gene polymorphism with idiopathic premature ovarian failure

To assess the influence of length of the alanine tract of forkhead box E1 (FOXE1) gene on genetic susceptibility to idiopathic premature ovarian failure (POF). Totally 110 patients with idiopathic POF were recruited between February 2009 and December 2012 at the Affiliated Shenzhen City Maternity and Child Healthcare Hospital of Southern Medical University. Controls (n = 110) were individuals with normal menstrual cycles, normal FSH concentrations. The polyalanine tract and flanking sequence of FOXE1 were screened using the multiplex ligation-dependent probe amplification (MLPA) technique and direct sequence technique. The most frequent of FOXE1 polyalanine stretch length was 14 residues in both groups. The length of FOXE1 polyalanine reported in this study varied from 12 to 16 alanines, and three variants of FOXE1-polyalanine length, containing 12, 14, or 16 alanine residues, and 5 different genotypes were identified. The most common genotypes were 14/14 homozygote, occurring with the frequency of 81.8% (90/110) in the POF group, while 96.4% (106/110) in control subjects, respectively. The incidence of 14/14 genotypes of FOXE1-polyalanine was significantly lower in patients with POF (χ² = 119.730, P = 0.001) in comparison to the controls. There were significantly higher frequencies of the 16/16 genotypes in cases with POF [10.0% (11/110) versus 0; χ² = 3.403, P = 0.001], as compared with the controls. The FOXE1 14 alanine allele was significantly less common in the POF patient group than the controls [84.5% (186/220) versus 98.2% (216/220); χ² = 25.923, P = 0.001]. The FOXE1 16 alanine allele was significantly more common in the POF patient group than the controls [12.7% (28/220) versus 1.8% (4/220); χ² = 19.412, P = 0.001]. The polymorphism of the polyalanine tract of FOXE1 gene have a certain relevance for the genetic aetiology of idiopathic POF.

Analysis of CGG repeats in FMR1 in Chinese women with idiopathic premature ovarian failure

Excessive triple CGG repeats in the FMR1 gene have been widely associated with premature ovarian failure. The number of AGG interruptions and length of uninterrupted CGG repeats have been correlated with repeat instability on transmission. In this study, FMR1 CGG repeats and AGG interruption status were determined by triplet-primed PCR in 117 premature ovarian failure patients and 82 matched controls. A possible relationship between CGG repeats or AGG interruption and serum FSH concentrations in patients and controls was evaluated. One patient had a premutation allele (73 repeats) (1/117), while no such mutations were observed in controls (0/82). Other patients and all controls had CGG repeats in the normal range. There was no significant difference in the incidence of intermediate mutations of CGG repeats between patients and controls and no relationship between CGG repeats with serum FSH concentrations. Interestingly, more individuals with premature ovarian failure carried no AGG interruptions than the controls (4.27% versus 1.83%) but statistical significance was not reached. This small case–control study failed to find associations between CGG repeat sizes or AGG interruptions in FMR1 and premature ovarian failure in Chinese women. Further study with large sample size is warranted.

Novel NR5A1 Missense Mutation in Premature Ovarian Failure: Detection in Han Chinese Indicates Causation in Different Ethnic Groups

BackgroundThe etiology of most premature ovarian failure (POF) cases is usually elusive. Although genetic causes clearly exist and a likely susceptible region of 8q22.3 has been discovered, no predominant explanation exists for POF. More recently, evidences have indicated that mutations in NR5A1 gene could be causative for POF. We therefore screened for mutations in the NR5A1 gene in a large cohort of Chinese women with non-syndromic POF.MethodsMutation screening of NR5A1 gene was performed in 400 Han Chinese women with well-defined 46,XX idiopathic non-syndromic POF and 400 controls. Subsequently, functional characterization of the novel mutation identified was evaluated in vitro.ResultsA novel heterozygous missense mutation [c.13T>G (p.Tyr5Asp)] in NR5A1 was identified in 1 of 384 patients (0.26%). This mutation impaired transcriptional activation on Amh, Inhibin-a, Cyp11a1 and Cyp19a1 gene, as shown by transactivation assays. However, no dominant negative effect was observed, nor was there impact on protein expression and nuclear localization.ConclusionsThis novel mutation p.Tyr5Asp, in a novel non-domain region, is presumed to result in haploinsufficiency. Irrespectively, perturbation in NR5A1 is not a common explanation for POF in Chinese.

Application of DHPLC screening TGFBR-3 gene in Chinese women with idiopathic premature ovarian failure

To evaluate clinical value of denaturing high performance liquid chromatography (DHPLC) used in detecting transforming growth factor beta receptor 3 (TGFBR-3) exons 11 and 12 polymorphism in women with idiopathic premature ovarian failure (POF). From Feb. 2009 to Dec. 2011, 110 patients with idiopathic POF undergoing treatment at Shenzhen Maternal & Child Health Institute affiliated to Southern Medical University were enrolled as POF group in this study. In the mean time, 110 women under 40 years old with normal hormonal level and menstrual cycles as control group. The exons 11 and 12 of TGFBR-3 gene polymorphism were screened by using DHPLC, and results of DNA sequencing was as golden standard. Some related indexes were calculated, such as sensitivity, specificity, false negative value, false positive value, Youden index, positive predictive value, and negative predictive value. At the same time, 20% of the tested specimens were chosen randomly and detected by DHPLC again. The value of Kappa index were calculated by comparing the results between the first and second DHPLC analysis. The exon 11 of TGFBR-3 were not identified gene polymorphism and two nucleotide polymorphisms were identified in exon 12. For 2022 T/C polymorphism, the frequencies of CC with 0.9% (1/110), TC with 22.7% (25/110), TT with 76.4% (84/110), C with 12.3% (27/220) and T with 87.7% (193/220) in POF group were significantly different from CC with 0, TC with 9.1% (10/110) and TT with 90.9% (100/110), C with 4.5% (10/220) and T with 95.5% (210/220) in control group (all P<0.05). Allelic and genotypic frequencies of 2161-75 C/T were not differed significantly between the two groups (all P>0.05). As DNA sequencing as golden standard, DHPLC showed that the sensitivity was 100%, specificity was 97.9%, Youden index was 97.9%, positive predictive value was 96.3%, negative predictive value was 100%, and Kappa index was 0.888 (P<0.05). DHPLC analysis is higher validity, reliability and practicability method in detecting TGFBR-3 polymorphism in idiopathic premature ovarian failure.