Investigation of the molecular genetic causes of non-syndromic primary ovarian ınsufficiency by next generation sequencing analysis

ABSTRACT Objective: The aim of this study is to investigate the molecular genetic causes of non-syndromic primary ovarian insufficiency (POI) cases with the gene panel based on next generation sequencing analysis and to establish the relationship between genotype and phenotype. Subjects and methods: Twenty three cases aged 14-40 years followed up with POI were included. Patients with a karyotype of 46, XX, primary or secondary amenorrhea before the age of 40, with elevated FSH (>40 IU/mL) and low AMH levels (<0.03 ng/mL) were included in the study. Molecular genetic analyzes were performed by the next generation sequencing analysis method targeted with the TruSightTM Exome panel. Results: Median age of the cases was 17.8 (14.0-24.3) years, and 12 (52%) cases admitted before the age of 18. Fifteen (65%) patients had consanguineous parents. In 2 (8.6%) cases, variants detected were in genes that have been previously proven to cause POI. One was homozygous variant in FIGLA gene and the other was homozygous variant in PSMC3IP gene. Heterozygous variants were detected in PROK2, WDR11 and CHD7 associated with hypogonadotropic hypogonadism, but these variants are insufficient to contribute to the POI phenotype. Conclusion: Genetic panels based on next generation sequencing analysis technologies can be used to determine the molecular genetic diagnosis of POI, which has a highly heterogeneous genetic basis.

Chromosomal abnormalities detected by karyotyping among patients with secondary amenorrhea: a retrospective study

ABSTRACT BACKGROUND: Chromosomal abnormalities (CAs) have been described in patients with secondary amenorrhea (SA). However, studies on this association are scarce. OBJECTIVES: To evaluate the frequency and types of CAs detected by karyotyping in patients with SA. DESIGN AND SETTING: This retrospective study was performed in a reference clinical genetic service in South Brazil. METHODS: Data were obtained from the medical records of patients with SA who were evaluated between 1975 and 2022. Fisher's bicaudate exact test and Student's t-test were used, and P < 0.05 was considered significant. RESULTS: Among 43 patients with SA, 14 (32.6%) had CAs, namely del (Xq) (n = 3), 45,X (n = 2), 46,X,r(X)/45,X (n = 2), 46,XX/45,X (n = 1), 46,X,i(q10)/45,X (n = 1), 47,XXX (n = 1), 46,XX/47,XXX (n = 1), 46,XX/47,XX,+mar (n = 1), 45,XX,trob(13;14)(q10;q10)/46,XXX,trob(13;14)(q10;q10) (n = 1), and 46,XX,t(2;21)(q23;q11.2) (n = 1). Additional findings were observed mostly among patients with CA compared with those without CA (P = 0.0021). No difference in the mean age was observed between the patients with SA with or without CAs (P = 0.268025). CONCLUSIONS: CAs are common among patients with SA, especially those with short stature and additional findings. They are predominantly structural, involve the X chromosome in a mosaic, and are compatible with the Turner syndrome. Patients with SA, even if isolated, may have CAs, particularly del (Xq) and triple X.

Effect of "Zhibian" (BL 54)-to-"Shuidao" (ST 28) needle insertion on the expression of Fas/FADD/Caspase-8 of death receptor pathway in rats with primary ovarian insufficiency / 中国针灸

OBJECTIVE@#To explore the effect of "Zhibian" (BL 54)-to-"Shuidao" (ST 28) needle insertion on the ovarian function in the rats with primary ovarian insufficiency (POI) and the potential effect mechanism based on the Fas/FADD/Caspase-8 of death receptor pathway.@*METHODS@#Forty-eight female SD rats were randomly divided into a blank group, a model group, a medication group and an acupuncture group, with 12 rats in each group. Except in the blank group, the rats in the other groups were intraperitoneally injected with cyclophosphamide to establish the POI model. In the acupuncture group, after successful modeling, the intervention was given with "Zhibian" (BL 54)-to- "Shuidao" (ST 28) needle insertion, once daily, 30 min in each intervention; and the duration of intervention was 4 weeks. In the medication group, estradiol valerate tablets were administered intragastrically, 0.09 mg•kg-1•d-1, for 4 weeks. The general situation and the estrous cycle of the rats were compared among groups. Using ELISA, the levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) in the serum were detected. HE staining was adopted to observe the morphological changes of ovarian tissue of rats. The protein expression of Fas, FADD and Caspase-8 in ovarian tissue was detected with immunohistochemistry and Western blot.@*RESULTS@#After modeling, except the rats of the blank group, the rats of the other groups had dry fur, lost hair, low spirits, reduced food intake, increased urination and loose stool. After intervention, the stool became regular gradually in the acupuncture group and the medication group. The percentage of estrous cycle disturbance was increased in the rats of the model group when compared with the blank group (P<0.01); in comparison with the model group, the percentages of estrous cycle disturbance were reduced in the acupuncture group and the medication group after intervention (P<0.01). When compared with the blank group, the body mass and E2 content in the serum were lower (P<0.01), the levels of FSH and LH in the serum and the protein expression levels of Fas, FADD and Caspase-8 were increased (P<0.01) in the model group. Compared with the model group, the body mass and E2 contents in the serum were higher (P<0.01), the levels of FSH and LH in the serum and the protein expression levels of Fas, FADD and Caspase-8 were reduced (P<0.01) in the acupuncture group and the medication group.@*CONCLUSION@#"Zhibian" (BL 54)-to-"Shuidao" (ST 28) needle insertion can effectively improve the ovarian function of POI rats, and its effect mechanism may be related to regulating the serum sex hormone levels, reducing the expression of Fas, FADD and Caspase-8 in ovarian tissue and retarding apoptosis of ovarian cells.

FMR1 alleles in women with idiopathic infertility / Alelos do gene FMR1 em mulheres com infertilidade idiopática

INTRODUCTION: The frequency of the premutation alleles of the FMR1 gene varies from 1:100 to 1:260 Israeli, Canadian, Finnish and American women, but it is unknown in Brazil. Premutation carriers may have reduced reproductive age and are at risk of transmitting the expanded allele to their offspring, and consequently fragile X syndrome. OBJECTIVE: To observe the distribution range of the FMR1 gene alleles in a population of women with idiopathic infertility, without symptoms of premature ovarian insufficiency. METHODS: The presence of premutation in FMR1 was assessed by conventional PCR, agarose, and acrylamide gel and analysis of fragments in capillary electrophoresis. Lymphocyte DNA obtained from 283 women undergoing infertility treatment was analyzed. RESULTS: 169 patients had the normal heterozygous allele (59.7%), 114 had the normal homozygous allele (40.6%) and no patient had the premutation. Premature ovarian insufficiency is seen in 20 to 30% of women with the permutated allele. Thus, the condition can be asymptomatic in a large part of the premutation carriers. Brazil has a diverse population and, therefore, the allele frequencies of many gene variants are unknown. Previous Brazilian studies have shown a low frequency of the premutation allele in different patient cohorts. Corroborating these articles, the results demonstrated that the frequency of the premutation allele is low in the infertile women population studied. CONCLUSION: Tracking the size of the FMR1 gene alleles allows the expansion of knowledge about the frequency of risk alleles associated with genetic diseases in the Brazilian population.

INTRODUÇÃO: A frequência dos alelos pré-mutados do gene FMR1 varia de 1:100 e 1:260 mulheres israelenses, canadenses, finlandesas e americanas, mas é desconhecida no Brasil. Portadoras da pré-mutação podem apresentar redução da idade reprodutiva e possuem risco de transmissão do alelo expandido para a prole, e consequentemente a Síndrome do X frágil. OBJETIVO: Observar a faixa de distribuição dos alelos do gene FMR1 em uma população de mulheres com infertilidade idiopática, sem sintomas de insuficiência ovariana prematura. MÉTODOS: A presença da pré-mutação em FMR1 foi avaliada por PCR convencional, gel de agarose e acrilamida e análise de fragmentos em eletroforese capilar. Analisou-se DNA de linfócitos obtidos de 283 mulheres em tratamento de infertilidade. RESULTADOS: Foi observado que 169 pacientes apresentam o alelo heterozigoto normal (59,7%), 114 apresentam o alelo homozigoto normal (40,6%) e nenhuma paciente apresentou a pré-mutação. A insuficiência ovariana prematura é observada em 20 a 30% das mulheres portadoras do alelo pré-mutado. Assim, a presença de um alelo pré-mutado pode ser assintomática em grande parte dos casos. O Brasil possui uma população diversificada e, portanto, as frequências alélicas de muitas variantes gênicas são desconhecidas. Estudos brasileiros anteriores mostraram uma baixa frequência do alelo pré-mutado em diferentes coortes de pacientes. Corroborando estes autores, os resultados demonstram que frequência do alelo pré-mutado é baixa na população de mulheres inférteis estudada. CONCLUSÃO: O rastreamento do tamanho dos alelos do gene FMR1 permite ampliar o conhecimento sobre a frequência dos alelos de risco para doenças genética na população brasileira.

Nueva variante del gen STAG3 causante de insuficiencia ovárica prematura / New STAG3 gene variant as a cause of premature ovarian insufficiency

RESUMEN Objetivos: describir un caso de falla ovárica secundaria a una variante patogénica homocigota en el gen STAG3 no reportada previamente. Materiales y métodos: paciente de 16 años con amenorrea primaria y ausencia de características sexuales secundarias, en quien se documentó hipotiroidismo autoinmune, pobre desarrollo genital y cintilla gonadal, por lo cual se realizó secuenciación de exorna clínico. Se identificó una variante homocigota patogénica previamente no reportada en el gen STAG3, el cual ha sido relacionado con insuficiencia ovárica prematura (IOP). Conclusiones: en este caso, la realización de exorna clínico fue determinante para identificar una alteración del gen STAG, probablemente asociada a la IOP y el pronóstico a largo plazo de la paciente. Se establece una nueva variante patogénica c.2773delT; p.Ser925Profs*6 del gen STAG3 asociada a la IOP.

ABSTRACT Objectives: To describe a case of ovarian failure secondary to a homozygous pathogenic variant in the STAG3 gene not previously reported. Material and methods: A 16-year-old patient with primary amenorrhea and absence of secondary sexual characteristics, with documented autoimmune hypothyroidism, poor genital and gonadal streak development which prompted the performance of clinical exorne sequencing. A homozygous pathogenic variant not previously reported in the STAG3 gene, which has been associated with premature ovarian insufficiency (POI), was identified. Conclusions: In this case, clinical exorne sequencing was key for identifying a STAG gene abnormality, probably associated with POI and long term prognosis for the patient. A new pathogenic variant c.2773delT; p.Ser925Profs*6 of the STAG3 gene associated with POI was established.

Insuficiencia ovárica primaria en adolescentes: revisión de la literatura a propósito de un caso clínico / Primary ovarian insufficiency in adolescents: review of the literature regarding a clinical case

El objetivo de este manuscrito es realizar una revisión y actualización de la literatura de la insuficiencia ovárica primaria (IOP) en población adolescente, a partir del diagnóstico, manejo y seguimiento de un caso clínico. La insuficiencia ovárica primaria se define como la menopausia en una mujer antes de los 40 años, acompañada de amenorrea, hipogonadismo hipergonadotrópico e infertilidad. Su prevalencia varía entre 1 a 2%, y en mujeres menores de 20 años su prevalencia es un caso de cada 10,000. Aunque se sabe que muchas afecciones pueden llevar a una IOP, la más común es la causa idiopática. La presentación clínica es diversa, y varios trastornos diferentes pueden también, llevar a esta condición. CASO CLÍNICO: Se presenta el caso de una adolescente de 17 años, previamente sana, con historia de amenorrea secundaria, no embarazada, con examen físico general y ginecológico normal. Se solicita estudio analítico complementario resultando con niveles de hormona folículo estimulante (FHS), estradiol (E2) y hormona antimülleriana (AMH) compatibles con una insuficiencia ovárica como la observada en la posmenopausia. Se inicia terapia hormonal (TH) clásica con estradiol y progesterona, siendo posteriormente reemplazada por anticoncepción hormonal combinada (AHC) oral, coincidente con el inicio de vida sexual, con respuesta favorable y sangrados regulares. La IOP tiene graves consecuencias para la salud incluyendo trastornos psicológicos como angustia, síntomas depresivos o depresión, infertilidad, osteoporosis, trastornos autoinmunes, cardiopatía isquémica, y un mayor riesgo de mortalidad. La enfermedad de Hashimoto es el trastorno autoinmune más frecuente asociado a la IOP. Su tratamiento y diagnóstico deben establecerse de forma precoz para evitar consecuencias a largo plazo. La terapia con estrógenos es la base del tratamiento para eliminar los síntomas de la deficiencia de estrógenos, además de evitar las consecuencias futuras del hipogonadismo no tratado. También el manejo debe incluir los siguientes dominios: fertilidad y anticoncepción, salud ósea, problemas cardiovasculares, función psicosexual, psicológica y neurológica, informando a los familiares y a la paciente sobre la dimensión real de la IOP y la necesidad de tratamiento multidisciplinario en muchos casos. CONCLUSIÓN: El caso presentado, pese a ser infrecuente, permite abordar de manera sistematizada el diagnostico de IOP y evaluar alternativas de manejo plausibles para evitar graves consecuencias en la salud, así como conocer respuesta clínica y de satisfacción de la adolescente.

The objective of this manuscript is to review and update the literature on primary ovarian insufficiency (POI) in an adolescent population, based on the diagnosis, management and follow-up of a clinical case. Primary ovarian insufficiency is defined as menopause in a woman before the age of 40, accompanied by amenorrhea, hypergonadotropic hypogonadism, and infertility. Its prevalence varies between 1 to 2%, and in women under 20 years of age its prevalence is one case in every 10,000. Although it is known that many conditions can lead to POI, the most common is the idiopathic cause. The clinical presentation is diverse, and several different disorders can also lead to this condition. CLINICAL CASE: The case of a 17-year-old adolescent, previously healthy, with a history of secondary amenorrhea, not pregnant, with a normal general physical and gynecological examination is presented. A complementary analytical study is requested, resulting in levels of follicle stimulating hormone (FHS), estradiol (E2) and anti-müllerian hormone (AMH) compatible with ovarian insufficiency such as that observed in postmenopause. Classic hormonal therapy (HT) with estradiol and progesterone was started, later being replaced by combined hormonal contraception (CHC), coinciding with the beginning of sexual life, with a favorable response and regular bleeding. POI has serious health consequences including psychological disorders such as distress, depressive symptoms or depression, infertility, osteoporosis, autoimmune disorders, ischemic heart disease, and an increased risk of mortality. Hashimoto's disease is the most common autoimmune disorder associated with POI. Its treatment and diagnosis must be established early to avoid long-term consequences. Estrogen therapy is the mainstay of treatment to eliminate the symptoms of estrogen deficiency, in addition to avoiding the future consequences of untreated hypogonadism. Management should also include the following domains: fertility and contraception, bone health, cardiovascular problems, psychosexual, psychological and neurological function, informing family members and the patient about the real dimension of POI and the need for multidisciplinary treatment in many cases. CONCLUSION: The case, although infrequent, allows a systematic approach to the diagnosis of POI and evaluate plausible management alternatives to avoid serious health consequences, as well as to know the clinical response and satisfaction of the adolescent.

Progress in molecular mechanism and clinical application of primordial follicle activation / 中国医师杂志

With the continuous elucidation of the molecular mechanism of primordial follicle activation, clinical in vitro activation (IVA) has gradually become a clinical treatment for premature ovarian insufficiency (POI) and reduced ovarian reserve (DOR). This article will review the mechanism of phosphatase and tensin homolog (PTEN)/phosphatidylinositol-3-kinase (PI3K), hippo signal and mammalian target of rapamycin complex 1 (mTORC1) signal involved in the activation of primordial follicles, and the application of IVA in patients with POI and DOR.

Assessment of FMR1 triplet repeats in patients affected with mental retardation, fragile X syndrome and primary ovarian insufficiency

The CGG repeats in the FMR1 gene expand in patients with fragile X syndrome, fragile X-associated tremour/ataxia syndrome and fragile X-associated primary ovarian failure. In this study, the CGG repeats in the FMR1 gene were studied in 449 males and 207 females using traditional polymerase chain reaction and triplet repeat primed PCR methods, also 18 CVS samples (six males and 12 females) were tested for prenatal diagnosis. Further, methylation sensitive multiplexed ligation dependent probe amplification was performed on some samples to confirm the results. Regarding the male patients, 1.1% and 9.7% had premutation (PM) and full mutation (FM) alleles, respectively. Also three (0.66%) male patients were mosaic for PM and FM alleles. Among females, 1.9% were GZ carriers and 5.8% were PM carriers. Prenatal diagnosis resulted in detection of two PM and one FM males as well as one FM carrier female. Our results were in concordance with the previously published results.

Sintomas geniturinários em mulheres com câncer de mama em tratamento quimioterápico / Genitourinary symptoms in women with breast cancer during chemotherapy treatment

Introdução: O câncer de mama representa a neoplasia mais incidente em mulheres, com projeção para representar 29% de todos os tipos de câncer para o biênio 2018-2019 no Brasil. A quimioterapia é a modalidade terapêutica mais utilizada para o tratamento desta neoplasia, utilizando-se largamente a droga ciclofosfamida, a qual apresenta diversos eventos adversos, dentre eles a supressão gônodal que pode induzir ou intensificar os sintomas menopausais. Objetivo: Analisar a qualidade de vida e ocorrência de sintomas geniturinários durante o tratamento quimioterápico adjuvante com os esquemas FAC; AC-T, CMF ou TC para câncer de mama. Método: Estudo prospectivo que avaliou mulheres antes (T0) e 30 dias após a realização do tratamento quimioterápico(T1). Foram avaliadas quanto ao status menopausal, a força e resistência exercida pelos músculos do assoalho pélvico com o perineômetro, além, de responderem a um questionário sociodemográfico, escala de avaliação da menopausa e o ICIQ-SF. Os dados foram analisados por meio de testes com qui-qradrado, teste t, análise multifatorial para medias repetidas, LSD-Fisher e análise de componentes principais. Resultados: A média de idade foi de 46,3 anos (DP± 5,77), as caraterísticas de maior prevalecia foram a hipertensão arterial (35%), carcinoma ducal infiltrante (90,2%), e estadiamento clinico em IIA (40%). Anterior a quimioterapia 60% das mulheres apresentavam ciclos menstruais regulares, mas nenhuma manteve a regularidade após quimioterapia tornaram-se amenorreicas. A escala de avaliação da menopausa apresentou piora significativa dos sintomas menopausais quando comparados T0 e T1(p<0,05) e quanto ao ICIQ-SF todas as mulheres apresentaram aumento significativo das médias do score final (p<0,001), demonstrando piora dos sintomas urinários e qualidade de vida. Os valores de contração rápida e lenta verificados com o perineômetro em T0 apresentaram redução significativa em T1 (p<0,05) e observou-se aumento da incidência de incontinência urinária de esforço. A análise dos componentes principais sintetizou os resultados mostrando que as variáveis obtidas pela perineometria, Escala de avaliação de menopausa ICIQ-SF não diferiam entre grupos de quimioterapia, status menopausal e número de partos (p>0,05), porém ao longo das avaliações, entre T0 e T1, houve diferença estatística (p<005). Conclusão: A exposição ao tratamento quimioterápico contendo ciclofosfamida, independente de outros fatores aqui avaliados, promove alterações geniturinárias

Introduction: Breast cancer represents the most common cancer in women, projected to represent 29% of all cancers for the 2018-2019 biennium in Brazil. Chemotherapy is the most widely used therapeutic modality for the treatment of this neoplasm, largely using the drug cyclophosphamide, which has several adverse events, including gonodal suppression that can induce or intensify menopausal symptoms. Objective: To analyze the quality of life and occurrence of genitourinary symptoms during adjuvant chemotherapy treatment with FAC regimens AC-T, CMF or CT for breast cancer. Method: Prospective study that evaluated women before (T0) and 30 days after chemotherapy (T1). They were evaluated for menopausal status, strength and endurance exerted by the pelvic floor muscles with the perineometer, and answered a sociodemographic questionnaire, menopause assessment scale and the ICIQ-SF. Data were analyzed using chi-square tests, t-test, repeated-factor multifactorial analysis, LSD-Fisher and principal component analysis. Results: The mean age was 46.3 years (SD ± 5.77), the most prevalent characteristics were arterial hypertension (35%), infiltrating ducal carcinoma (90.2%), and clinical staging in IIA (40%). Prior to chemotherapy 60% of women had regular menstrual cycles, but none maintained regularity after chemotherapy became amenorrheic. The menopause assessment scale showed a significant worsening of the menopausal symptoms when compared to T0 and T1 (p<0.05). Regarding the ICIQ-SF, all women presented a significant increase in the final score means (p<0.001), showing a worsening of the urinary symptoms and quality of life. The fast and slow contraction values verified with the perineometer at T0 showed a significant reduction at T1 (p<0.05) and an increased incidence of stress urinary incontinence was observed. The principal component analysis summarized the results showing that the variables obtained by perineometry, ICIQ-SF Menopause Rating Scale did not differ between chemotherapy groups, menopausal status and number of deliveries (p>0.05), but throughout the evaluations, between T0 and T1, there was a statistical difference (p<005). Conclusion: Exposure to chemotherapeutic treatment containing cyclophosphamide, regardless of other factors evaluated here, promotes genitourinary changes

Fragilidad del X y otras entidades asociadas al gen FMR1: estudio de 28 familias afectadas / Fragile X syndrome and other entities associated with the FMR1 gene: Study of 28 affected families

El síndrome de fragilidad del cromosoma X es la causa de discapacidad intelectual heredable más frecuente. Asociado a trastornos del espectro autista en un tercio de los pacientes, afecta, con mayor prevalencia, a los varones. Se debe a una expansión de trinucleótidos CGG (citosina, guanina, guanina), llamada mutación completa en el locus Xq27.3 del gen FMR1, que conduce a la hipermetilación en el promotor del gen y reduce los niveles de expresión de FMRP, una proteína implicada en la maduración y plasticidad sináptica. Una expansión menor de CGG es la causa de insuficiencia ovárica primaria y del síndrome de temblor/ataxia asociado a X frágil, caracterizado por ataxia cerebelosa progresiva, de inicio tardío, y temblor de intención. En el presente estudio de serie de casos, se analiza la segregación de mutaciones del gen FMR1 en diferentes familias y la variabilidad de expresión clínica que llevó a la consulta genética.

The fragile X syndrome occurs due to an expansion of CGG trinucleotides, called full mutation, which is found at the Xq27.3 locus of the FMR1 gene. It is the most common cause of inherited intellectual disability. Associated with autistic spectrum disorders in one third of the patients, it affects males with higher prevalence. It also leads to hypermethylation of the gene promoter, silencing it and reducing the expression levels of FMRP, a protein involved in synaptic maturation and plasticity. A lower expansion causes primary ovarian failure syndrome as well as tremor and ataxia syndrome characterized by progressive cerebellar ataxia of late onset and intention tremor. In the present case-control study we analyze the segregation of mutations of the FMR1 gene in different families and the variability of expression that led to the genetic consultation.

Effect of Bushen Yanggan decoction combine with acupoint catgut embedding therapy in the treatment of patients with premature ovarian failure and its influence on serum follicle stimulating hormone and estradiol levels / 中国基层医药

Objective To study the effect of Bushen Yanggan decoction combine with acupoint catgut embedding therapy in the treatment of patients with premature ovarian failure and its influence on serum follicle stimulating hormone(FSH) and estradiol(E2) levels. Methods From January 2016 to June 2017,76 patients with premature ovarian failure in the First People＇s Hospital of Xiaoshan District were selected. They were divided into observation group(n=38) and control group(n=38) by the random number table method. The control group was given western medicine hormone replacement therapy, the observation group was given Bushen Yanggan decoction combined with acupoint catgut embedding therapy. The patients in the two groups were treated for 3 months. The clinical efficacy,clinical symptoms,serum FSH,E2levels and endometrial thickness before and after treatment were compared between the two groups. Results The total effective rate of the observation group was 86. 84% (33/38), which was slightly higher than 78. 95% (30/38) of the control group,but the difference was not statistically significant (χ2=0. 835,P＞0. 05). The incidence rates of soreness and weakness of waist and knees,hot flashes sweating,sexual decline,loss of control of the observation group after 3 months of treatment were 21. 05% (8/38),26. 32% (10/38), 18.42%(7/38),21. 05% (8/38),respectively,which were significantly lower than those of the control group [44.74%(17/38),50. 00% (19/38),39. 47% (15/38),42. 11% (16/38)],the differences were statistically significant(χ2=4. 828,4. 517,4. 094,3. 897,all P＜0. 05). The serum FSH of the observation group after 3 months of treatment was (31. 44 ± 11. 04) IU/L,which was lower than (60. 38 ± 13. 02) IU/L of the control group,the difference was statistically significant(t =10. 447,P ＜0. 05). The serum E2levels,endometrial thickness of the observation group were (92. 77 ± 10. 72) pmol/L,(7. 95 ± 1. 17) mm,respectively,which were higher than those of the control group[(60.24±11.33)pmol/L,(4.82 ±1.65)mm],the differences were statistically significant(t=12. 856,9. 539,all P＜0. 05). Conclusion Bushen Yanggan decoction combined with acupoint catgut embedding therapy for premature ovarian failure has good curative effect,it can improve the clinical symptoms and serum sex hormone levels,increase the thickness of the endometrium,which is worthy of clinical application.

Meta-analysis on modified Zuogui Pill combined with hormone in the treatment of premature ovari-an failure / 中国医师杂志

Objective To evaluate the effectiveness and safety of modified Zuogui Pill combined with hormone in the treatment of premature ovarian failure ( POF) . Methods Randomized controlled trials were carried out by searching Chinese and English databases from the establishing to June, 2018. The meth-odological quality was assessed in the included randomized controlled trials. The treatment of premature o-varian failure by modified Zuogui Pill combined with hormone was systematically evaluated by RevMan 5. 3 software. Results 11 randomized controlled trials involving 1004 patients were included. Meta-analysis results showed that total effective rate (OR=4. 44, 95％ CI: 3. 10 -6. 36, P<0. 00001), Tranditional Chinese Medcine ( TCM ) syndromes ( including dizziness, tinnitus, hot flash night sweats, five upset fe-ver, waist and knees), serum follicle stimulating hormone ( FSH) level [ standardised mean difference (SMD) =0. 54, 95％ CI:0. 13-0. 95, P=0. 01], luteinizing hormone (LH) level (SMD=0. 90, 95％CI:0. 45-1. 36, P<0. 0001), estradiol (E2)level (SMD=1. 26, 95％ CI:0. 12-2. 40, P=0. 03), endometrial thickness [weighted mean difference (WMD) =2. 03, 95％ CI:0. 07-3. 99, P=0. 04] im-provement and the incidence of adverse reactions (OR=0. 34, 95％ CI:0. 12-0. 95, P=0. 04) in modi-fied Zuogui Pill combined with hormone therapy were superior to those of pure hormone therapy. Conclu-sions The modified Zuogui Pill combined with hormone therapy shows better therapeutic effect and safety than hormone therapy on premature ovarian failure.

Primordial follicle activation as new treatment for primary ovarian insufficiency / 대한생식의학회지

Primordial follicle activation is a process in which individual primordial follicles leave their dormant state and enter a growth phase. While existing hormone stimulation strategies targeted the growing follicles, the remaining dormant primordial follicles were ruled out from clinical use. Recently, in vitro activation (IVA), which is a method for controlling primordial follicle activation, has provided an innovative technology for primary ovarian insufficiency (POI) patients. IVA was developed based on Hippo signaling and phosphatase and tensin homolog (PTEN)/phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT)/forkhead box O3 (FOXO3) signaling modulation. With this method, dormant primordial follicles are activated to enter growth phase and developed into competent oocytes. IVA has been successfully applied in POI patients who only have a few remaining remnant primordial follicles in the ovary, and healthy pregnancies and deliveries have been reported. IVA may also provide a promising option for fertility preservation in cancer patients and prepubertal girls whose fertility preservation choices are limited to tissue cryopreservation. Here, we review the basic mechanisms, translational studies, and current clinical results for IVA. Limitations and further study requirements that could potentially optimize IVA for future use will also be discussed.

Consequences of chemotherapeutic agents on primordial follicles and future clinical applications

The ovarian reserve is necessary for female fertility and endocrine health. Commonly used cancer therapies diminish the ovarian reserve, thus, resulting in primary ovarian insufficiency, which clinically presents as infertility and endocrine dysfunction. Prepubertal children who have undergone cancer therapies often experience delayed puberty or cannot initiate puberty and require endocrine support to maintain a normal life. Thus, developing an effective intervention to prevent loss of the ovarian reserve is an unmet need for these cancer patients. The selection of adjuvant therapies to protect the ovarian reserve against cancer therapies underlies the mechanism of loss of primordial follicles (PFs). Several theories have been proposed to explain the loss of PFs. The “burn out” theory postulates that chemotherapeutic agents activate dormant PFs through an activation pathway. Another theory posits that chemotherapeutic agents destroy PFs through an “apoptotic pathway” due to high sensitivity to DNA damage. However, the mechanisms causing loss of the ovarian reserve remains largely speculative. Here, we review current literature in this area and consider the mechanisms of how gonadotoxic therapies deplete PFs in the ovarian reserve.

Criopreserved ovarian tissue transplantation and bone restoration metabolism in castrated rats / Transplante de tecido ovariano criopreservado e restauração do metabolismo ósseo em ratas castradas

ABSTRACT Objectives: to evaluate estradiol levels and autotransplantation heated ovarian tissue effects, after vitrification, on rats bone metabolism previously oophorectomized bilaterally. Methods: experimental study with 27 rats aged 11 to 12 weeks and weighing 200g to 300g, submitted to bilateral oophorectomy and ovarian tissue cryopreservation for subsequent reimplantation. Animals were divided into two groups, A and B, with 8 and 19 rats, respectively. Autotransplantation occurred in two periods according to castration time: after one week, in group A, and after one month in group B. Serum estradiol measurements and ovary and tibia histological analysis were performed before and after oophorectomy period (early or late) and one month after reimplantation. Results: in groups A and B, tibia median cortical thickness was 0.463±0.14mm (mean±SD) at the baseline, 0.360±0.14mm after oophorectomy and 0.445±0.17mm one month after reimplantation p<0.005). Trabecular means were 0.050±0.08mm (mean±SD) at baseline, 0.022±0.08mm after oophorectomy and 0.049±0.032mm one month after replantation (p<0.005). There was no statistical difference in estradiol variation between the two study groups (p=0.819). Conclusion: cryopreserved ovarian tissue transplantation restored bone parameters, and these results suggest that ovarian reimplantation in women may have the same beneficial effects on bone metabolism.

RESUMO Objetivos: avaliar os níveis de estradiol e os efeitos do autotransplante de tecido ovariano aquecido, após vitrificação, no metabolismo ósseo de ratas previamente ooforectomizadas bilateralmente. Métodos: trabalho experimental com 27 ratas com idades entre 11 e 12 semanas e pesando 200g a 300g, submetidas à ooforectomia bilateral e criopreservação de tecido ovariano para posterior reimplante. Os animais foram divididos em dois grupos, A e B, com oito e 19 ratas, respectivamente. O autotransplante ocorreu em dois períodos de acordo com o tempo de castração: após uma semana, no grupo A, e após um mês no grupo B. Mensurações de estradiol sérico e análise histológica de ovário e tíbia foram feitos antes e após o período de ooforectomia (precoce ou tardio) e um mês após o reimplante. Resultados: nos grupos A e B, as espessuras corticais médias da tíbia foram 0,463±0,14mm (média±DP) na linha de base, 0,360±0,14mm após ooforectomia e 0,445±0,17mm em um mês após o reimplante (p<0,005). As médias trabeculares foram 0,050±0,08mm (média±DP) na linha de base, 0,022±0,08mm após ooforectomia e 0,049±0,032mm em um mês após o reimplante (p<0,005). Não houve diferença estatística entre a variação do estradiol entre os dois grupos de estudo (p=0,819). Conclusão: o transplante de tecido ovariano criopreservado restabeleceu os parâmetros ósseos, e estes resultados sugerem que a reimplantação ovariana em mulheres pode apresentar os mesmos efeitos benéficos sobre o metabolismo ósseo.

Pregnancy outcome of patients ≥ 35 years old with diminished ovarian reserve treated with growth hormone / 中国医师杂志

Objective To explore the application effect of growth hormone in patients ≥35 years old with diminished ovarian reserve (DOR),and to analyze pregnancy outcome.Methods A total of 120 cases with diminished ovarian reserve (≥35 years old) in our hospital were randomly divided into observation group and control group,each group of 60 cases.The control group was given conventional treatment,while the observation group added growth hormone.Changes of follicle stimulatng homone (FSH) and luteinizing hormone (LH),FSH/LH,serum estradiol (E2) level were compared between the two groups before and after treatment,along with the pregnancy outcome of two groups such as normal fertilization rate,excellent embryo rate,quantity of egg,cleavage rate,cycle cancellation rate.Results After treatment,the FSH and FSH/LH of the observation group were significantly lower than those of the control group,the serum E2 was significantly higher than that of the control group (P ＜ 0.05);the normal fertilization rate,cleavage rate and excellent embryo rate in the observation group were 53.96％,86.67％ and 35.38％ respectively,which were significantly higher than those of 33.58％,71.11％ and 15.63％ in the control group (P ＜ 0.05);the cycle cancellation rate of the observation group was 6.67％,which was significantly lower than that of 21.67％ in the control group (P ＜ 0.05);the biochemical pregnancy rate and clinical pregnancy rate of the observation group were 38.33％ and 28.33％,which were significantly higher than those of 20％ and 13.33％ in the control group (P ＜ O.05).Conclusions Diminished ovarian reserve in patients ≥35 years old with growth hormone therapy,can promote the improvement of ovarian function,the biochemical pregnancy rate and clinical pregnancy rate,it is worthy of clinical promotion.

Establishment of Effective Mouse Model of Premature Ovarian Failure Considering Treatment Duration of Anticancer Drugs and Natural Recovery Time

OBJECTIVES: This study was aimed to establish the most effective premature ovarian failure (POF) mouse model using Cyclophosphamide (CTX), busulfan (Bu), and cisplatin considering treatment duration of anticancer drugs and natural recovery time. METHODS: POF was induced by intraperitoneally injecting CTX (120 mg/kg)/Bu (12 mg/kg) for 1 to 4 weeks or cisplatin (2 mg/kg) for 3 to 14 days to C57BL/6 female mice aged 6 to 8 weeks. Controls were injected with equal volume of saline for the same periods. Body weight was measured every week, and ovarian and uterine weights were measured after the last injection of anticancer drug. To assess ovarian function, POF-induced mice were superovulated with pregnant mare serum gonadotropin and human chorionic gonadotropin, and then mated with male. After 18 hours, zygotes were retrieved and cultured for 4 days. Finally, the mice were left untreated for a period of times after the final injection of anticancer drug, and the time for natural recovery of ovarian function was evaluated. RESULTS: After 2 weeks of CTX/Bu injection, ovarian and uterine weights, and ovarian function were decreased sharply. Cisplatin treatment for 10 days resulted in a significant decrease in ovarian and uterine weight, and ovarian function. When POF was induced for at least 2 weeks for CTX/Bu and for at least 10 days for cisplatin, ovarian function did not recover naturally for 2 weeks and 1 week, respectively. CONCLUSIONS: These results suggest that CTX/Bu should be treated for at least 2 weeks and cisplatin for at least 10 days to establish the most effective primary ovarian insufficiency mouse model.

Update on 46,XX primary ovarian insufficiency in adolescents / 中华实用儿科临床杂志

46,XX primary ovarian insufficiency(POI)is a clinical syndrome defined by loss of ovarian activity before the age of 40 years old with a karyotype 46,XX,characterized by menstrual disturbance(amenorrhea or oligomenorrhea)in association with hypergonadotropic hypogonadism.46,XX POI is a rare disease with the prevalence lower than 1％,of whom 2.5％are adolescents.Potential etiologies for 46,XX POI can be divided into genetic,autoimmune,and iatrogenic categories.Unfortunately,for most patients presenting with POI,the cause will remain unexplained.Once,POI is diagnosed,clinical indicated tests are needed to identify the mechanism causing POI.POI is a complex condition appearing with a strong genetic basis.Large-scale genomic sequencing had recently identified new mechanisms of POI.The management of the condition should address both of physical and emotional well-being health,including health education,hormone develoment treatment,prevention and treament of lower-estrogen associated diseases,with the support from a multidisciplinary team.

Curative effect of Kuntai Capsules on POI rats induced by VCD / 中成药

AIM To investigate the curative effects of Kuntai Capsules (Coptidis Rhizoma,Scutellariae Radix,Asini Corii Colla,etc.) on rat model for primary ovarian insufficiency (POI) caused by 4-vinylcyclohexene diepoxide (VCD).METHODS Seventy-five weaned 21-day old rats were divided equally into five groups randomly:control group,model group,low-,medium-,and high-dosage Kuntain group.Except the control group was given sesame oil rats,other four groups were given intraperitoneal injection of VCD solution.According to vaginal smear of rats,eight rats in each group were selected,the levels of FSH (follicle-stimulating hormone),LH (leuteinizing hormone),E2 (estradiol) and AMH (anti-Mullerian hormone) were measured,and morphological changes of ovary and uterus were observed.RESULTS Compared to the control group,FSH level in rats' serum of the model group significantly increased while AMH and E2 levels significantly decreased,and LH level rose;Kuntai Capsules significantly decreased FSH level while increased AMH level;The wet weight of rats' ovary in the model group was lighter than that in the control group.The ovary weight of Kuntai Capsules low-dosage group was increased;the number of follicles at all levels and corpora lutea of rats in the model group were decreased.After the treatment with Kuntai Capsules,the number of follicles at all levels and corpora lutea were increased;light microscopy manifested decreased uterine glands of the tube wall,reduced glandular cavity.After the treatment with Kuntai Capsules,the number of uterine glands increased and the gland cavity expanded.CONCLUSION Kuntai Capsules may slowdown the functional decline rate of ovarian.

Effects of autologous blood injection at Zusanli (ST 36) on ovarian function in patients with primary ovarian insufficiency / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of autologous blood injection and 0.9% NaCl at Zusanli (ST 36) on ovarian function in patients with primary ovarian insufficiency.</p><p><b>METHODS</b>Sixty patients with primary ovarian insufficiency were randomly divided into an observation group and a control group, 30 cases in each one. The patients in the observation group were treated with injection of autologous blood at Zusanli (ST 36); the patients in the control group were treated with 0.9% NaCl with identical volume at Zusanli (ST 36). Both the treatments were given once a week for 3 months. The ovarian function, including follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E) were tested before treatment, 1 month, 2 months and 3 months after first acupoint injection; the endometrial thickness before and after treatment and clinical efficacy were compared in the two groups.</p><p><b>RESULTS</b>Compared before treatment, FSH was lowered in the observation group after 1-month treatment (<0.05), while FSH and LH were lowered and Ewas increased after 2-month treatment and 3-month treatment (all<0.05). Compared with 1-month treatment, FSH and LH were lowered and Ewas increased in the observation group after 2-month treatment and 3-month treatment (all<0.05). Compared with 2-month treatment, FSH was lowered and Ewas increased in the observation group after 3-month treatment (both<0.05). The differences of all serum tests before and after treatment were insignificant in the control group (all>0.05). The FSH after 1-month treatment, and FSH, LH and Eafter 2-month treatment and 3-month treatment in the observation group were significantly different from those in the control group (all<0.05). The endometrial thickness after treatment in the observation group was higher than that before treatment (<0.05), while the endometrial thickness after treatment in the control group was similar to that before treatment (>0.05); the difference of endometrial thickness before and after treatment in the observation group was higher than that in the control group (<0.05). The clinical effective rate was 83.3% (25/30) in the observation group, which was superior to 46.7% (14/30) in the control group (<0.05).</p><p><b>CONCLUSION</b>The autologous blood injection at Zusanli (ST 36) can significantly improve ovarian function, promote endometrial growth in patients with primary ovarian insufficiency.</p>