Progression of Tripterygium wilfordii polyglycoside in mice with premature ovarian insufficiency at various time points / 中国比较医学杂志

Objective Changes in relevant indexes in the mouse model of early-onset ovarian insufficiency caused by Tripterygium wilfordii polyglycoside were analyzed,and the optimal time point for intervention was determined.Methods Forty female ICR mice were randomly divided into control and A,B,C,and D model groups with eight mice in each group.The control group was gavaged with purified water for 14 days(0.01 mL/10 g),and the remaining groups were administered a Tripterygium wilfordii polyglycoside suspension(80 mg/kg,0.01 mL/10 g)for 1 day(A model group),3 days(B model group),7 days(C model group),or 14 days(D model group),and samples were collected.Body weight and wet weights of the uterus and bilateral ovaries of mice were determined in each group.Serum FSH,LH,E2,P,AMH,INH-B,and T contents were measured using enzyme-linked immunoassays.HE staining was used to observe the number and developmental status of follicles and corpus luteum at all levels in mice of each group.TUNEL staining was used to detect the apoptosis in the ovaries of mice in each group.IHC detected expression of VEGFA,CD34,and EPO proteins in the ovaries of mice in each group.mRNA expression of HIF-1α,SDF-1,and CXCR4 in each group of mice was detected by PCR.Results Compared with the control group,changes in indicators in model A mice did not meet the POI modeling standard.The ovarian index,uterine index,and body weight of mice in the B model group were decreased significantly(P＜0.01),the weight of the C model group was decreased significantly(P＜0.01),and the ovarian index of the D model group was decreased significantly(P＜0.05).Serum contents of FSH and LH in B,C,and D model groups were increased(P＜0.05,P＜0.01),the E2,PROG,AMH,INH-B,and T contents were decreased(P＜0.01).The numbers of basal follicles,pre-sinus follicles,sinusoidal follicles,antral follicles,preovulatory follicles,and corpus luteum were decreased significantly(P＜0.05,P＜0.01)and the number of atresia follicles was increased significantly(P＜0.01)in B,C,and D model groups.The apoptotic area of TUNEL staining in A,B,C,and D model groups was increased significantly(P＜0.05,P＜0.01).Expression of CD34,VEGFA,and EPO in B,C,and D model groups was decreased significantly(P＜0.05,P＜0.01).mRNA expression of HIF-1α,SDF-1,and CXCR4 in A and B model groups was significantly increased(P＜0.05,P＜0.01).Compared with the B model group,the relevant indexes of C and D model groups were changed significantly,indicating that C and D models were more serious and tended to develop POF.Conclusions The B model group is the turning point of ovarian function from impaired POI to irreversible POF,suggesting that 3 days of administrating Tripterygium wilfordii polyglycoside is optimal to induce a POI disease model for effective drug intervention.

Study on the Mechanism of Guanyuan Mingmen Sequential Acupuncture Activating FSHR/cAMP/PKA Pathway to Promote Granulosa Cell Proliferation in POI Model Rats / 广州中医药大学学报

Objective To observe the therapeutic effects and mechanisms of Guanyuan Mingmen Sequential Acupuncture on rats with premature ovarian insufficiency(POI)model.Methods Female SD rats were divided into the blank group,the model group,the protein kinase A(PKA)inhibitor(H89)+acupuncture group,and the acupuncture group,with 12 rats in each group.Except for the blank group,the POI model was prepared by gavage with Tripterygium Glycosides Tablets in the other three groups of rats.After the model was successfully established,the blank group and the model group were bundled once a day;in the acupuncture group,Guanyuan(RN4)point was taken during the intermotility period,and in the pre-motility period,Mingmen(DU4)point was taken;in the H89+acupuncture group,the intervention was performed in accordance with the acupuncture protocol of the acupuncture group,and H89 was injected intraperitoneally for 30 minutes prior to each acupuncture session.Continuous intervention was performed for 20 days.Samples were taken from each group of rats in the first estrus period and in proestrus period after intervention.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of follicle stimulating hormone(FSH)and estradiol(E2)during the estrous phase,Western Blot was used to measure the protein expressions of follicle stimulating hormone receptor(FSHR)and aromatase P450(P450arom)during the estrous phase,and the activity of granulocytes during the estrous phase and the proestrus phase were measured using the cell-counting kit 8(CCK-8)method.The immunohistochemistry method was used to detect the protein expression of pre-motility proliferating cell nuclear antigen(PCNA).Results(1)Compared with the blank group,the serum FSH level of the model group and H89+acupuncture group was significantly increased(P＜0.01),and the E2 level was significantly decreased(P＜0.001);there was no difference between the FSH level of the H89+acupuncture group and that of the model group(P＞0.05),and the E2 level of the H89+acupuncture group was lower than that of the model group(P＜0.05);the FSH level of the acupuncture group was lower than that of the model group and that of the H89+acupuncture group(P＜0.05),had no difference with the blank group(P＞0.05),E2 level was significantly higher than the model group and H89+ acupuncture group(P＜0.01),still being lower than the blank group(P＜0.05).(2)The protein expressions of FSHR and P450arom in the model group and H89 + acupuncture group was lower than those in the blank group;the protein expression of FSHR in the H89 + acupuncture group was not different from that in the model group(P＞0.05),while the protein expression level of P450arom was lower than that in the model group(P＜0.05);the protein expression levels of FSHR and P450arom in the acupuncture group were higher than those in the model group and H89 + acupuncture group,but still lower than those in the blank group(P＜0.05).(3)Both GCs activity and average optical density value of PCNA in the model group and H89+acupuncture group were lower than the blank group(P＜0.05);both GCs activity and average optical density value of PCNA in the H89+acupuncture group were lower than the model group(P＜0.05);the activity of GCs and average optical density value of PCNA of the acupuncture group were significantly higher than that of the model group and H89+acupuncture group(P＜0.05 or P＜0.01).Conclusion Guanyuan Mingmen Sequential Acupuncture can regulate sex hormone levels,increase GCs activity and promote GCs cell proliferation by up-regulating protein expressions of follicle stimulating hormone receptor(FSHR)/cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)pathway FSHR,P450arom,thus improving POI.

The molecular mechanism of Liuwei Dihuang Pill in the treatment of premature ovarian insufficiency in a mouse model / 中国组织工程研究

BACKGROUND:Most of the formulas for the clinical treatment of premature ovarian insufficiency have evolved from the basic formula of Liuwei Dihuang Pills,and have achieved good therapeutic efficacy.Currently,most of the experimental studies on Liuwei Dihuang Pills focus on morphological observations and physiological and biochemical detection of in vivo animal models,while fewer studies on molecular mechanisms have been reported. OBJECTIVE:To explore the molecular mechanism of Liuwei Dihuang Pills in the treatment of premature ovarian insufficiency based on the receptor gamma coactivator-1 alpha/mitochondrial transcription factor A/reactive oxygen species pathway. METHODS:Premature ovarian insufficiency model was established in mice by intraperitoneal injection of cyclophosphamide 120 mg/kg combined with busulfan 12 mg/kg,and then Liuwei Dihuang Pill suspension was used to intervene in premature ovarian insufficiency mice.After 12 weeks of intervention,the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,anti-Mullerian hormone,8-hydroxydeoxyguanosine,total antioxidant capacity and reactive oxygen species in serum of mice were detected by ELISA method.The morphological changes in mouse ovaries were observed by hematoxylin-eosin staining.The ultrastructure of mouse follicular granulosa cells and the apoptosis of granulosa cell mitochondria were observed by transmission electron microscopy.The expression levels of receptor gamma coactivator-1 alpha and mitochondrial transcription factor A in mouse ovarian granulosa cells were detected by immunohistochemistry. RESULTS AND CONCLUSION:Compared with the model group,serum levels of follicle-stimulating hormone,luteinizing hormone,reactive oxygen species,and 8-hydroxydeoxyguanosine were decreased in the experimental group(P＜0.05),and the levels of estradiol,anti-Mullerian hormone,and total antioxidant capacity were increased(P＜0.05).Hematoxylin-eosin staining showed that in the model group,there were more atretic follicles and corpus luteum forms,some secondary follicles,and interstitial fibrosis and hyperplasia;in the experimental group,a large number of atretic follicles,few corpus luteum forms,primordial follicles were observed at the edges but there were few secondary follicles and no mature follicles.Transmission electron microscopy showed that the organelles in ovarian granulosa cells of mice in the experimental groups were relatively intact.Immunohistochemical results showed that compared with the model group,the expression level of receptor gamma coactivator-1 alpha in the ovarian tissue of mice increased slightly in the experimental group at the 4th week,and there was no significant change at the 8th and 12th weeks.The expression level of mitochondrial transcription factor A in the ovarian tissues of mice in the experimental group was transiently increased at the 4th week,and then slightly decreased,which were all significantly different from those of the model group.To conclude,Liuwei Dihuang Pills inhibit ovarian granulosa cell apoptosis in mice with premature ovarian insufficiency to a certain extent through the receptor gamma coactivator-1 alpha/mitochondrial transcription factor A/reactive oxygen species signaling pathway,thereby improving the endocrine function of the ovary,enhancing the antioxidant capacity,and attenuating the degree of oxidative stress damage.

Myasthenia Gravis with subsequent premature ovarian insufficiency: A case report

@#The incidence rate of Myasthenia Gravis coexisting with other autoimmune diseases is approximately 8.7 – 25%, but it is rarely associated with premature ovarian insufficiency (POI) with only less than 1% of women affected. This is a case of premature ovarian insufficiency in a 29 year old woman diagnosed with Myasthenia Gravis, who presented with lower extremity weakness and experienced two episodes of myasthenic crisis requiring thymectomy. Three years after, she noted oligomenorrhea that quickly progressed to amenorrhea. Extensive immunologic and genetic investigative studies showed no identifiable cause for the POI, except for its close temporal relationship with the occurrence of Myasthenia Gravis. The patient has been responsive to hormone replacement and immunomodulation therapy, and has not developed any further episodes of myasthenic crisis. A review of seven other reported cases describing a similar condition was also included in the discussion.

Xinjia Congrong Tusizi Decoction Regulates Ferroptosis of Granulosa Cells in Rat Model of Premature Ovarian Insufficiency via p53/Nrf2 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effects of Xinjia Congrong Tusizi decoction （XJCTD） on ovarian functions in the rat model of premature ovarian insufficiency （POI） and decipher the mechanism of regulating the tumor suppressor protein （p53）/nuclear factor E2-related factor 2 （Nrf2） pathway to attenuate granulosa cell ferroptosis. MethodForty-eight SPF-grade female SD rats were randomized into control， model， low-， medium-， and high-dose （1.1， 2.2， 4.4 g·kg-1） XJCTD， and Western medicine （coenzyme Q10， 0.002 7 g·kg-1） groups， with eight rats in each group. The rat model of POI was established by gavage of triptolide （TP）， and after successful modeling， each group was administrated with the corresponding drugs by gavage for 14 d. The body weight and ovarian weight of each rat were weighed and the ovarian index was calculated. The morphology of the ovarian tissue was observed by hematoxylin-eosin staining， and the proportions of growing follicles and atretic follicles were calculated. The serum levels of anti-Müllerian hormone （AMM）， estradiol （E2）， and follicle-stimulating hormone （FSH） were measured by enzyme-linked immunosorbent assay （ELISA）. The DCFH-DA fluorescent probe was used to measure the reactive oxygen species （ROS） content in granulosa cells. The content of cellular Ferrous ion （Fe2+）， lipid peroxide （LPO）， malondialdehyde （MDA）， glutathione （GSH）， and superoxide dismutase （SOD） was detected by colorimetry. The expression of the tumor suppressor protein p53，Nrf2， solute carrier family 7 member 11 （SLC7A11）， and glutathione peroxidase 4 （GPX4） was determined by immunohistochemistry and Western blot. ResultCompared with the control group， the model group showed decreased ovarian weight， body weight， and ovarian index （P<0.01）， reduced ovarian tissue volume and proportion of growing follicles （P<0.01）， increased proportion of atretic follicles （P<0.01）， lowered AMH and E2 levels and elevated FSH level in the serum （P<0.01）， and elevated levels of Fe2+， ROS， LPO， and MDA （P<0.01） and lowered levels of GSH and SOD in granulosa cells （P<0.01）. Moreover， the modeling up-regulated the expression of p53 （P<0.01） and down-regulated the expression of Nrf2， SLC7A11， and GPX4 （P<0.05， P<0.01） in the ovarian tissue. Compared with the model group， XJCTD increased the body weight， ovarian weight， and ovarian index （P<0.01）， alleviated the pathological changes in the ovarian tissue， increased the proportion of growing follicles （P<0.01）， decreased the proportion of atretic follicles （P<0.01）， and reduced the content of ROS in granulosa cells （P<0.05， P<0.01）. In addition， medium- and high-dose XJCTD lowered the FSH level （P<0.01） and raised E2 and AMH levels （P<0.01） in the serum， reduced the Fe2+ content （P<0.05， P<0.01）， and increased the SOD content （P<0.01） in granulosa cells. High-dose XJCTD reduced the LPO and MDA content （P<0.01） and increased the SOD content （P<0.01） in the granulosa cells， down-regulated the expression of p53 （P<0.05）， and up-regulated the expression of Nrf2， SLC7A11， and GPX4 in the ovarian tissue （P<0.05， P<0.01）. ConclusionXJCTD may protect the ovarian function in the rat model of POI by regulating the p53/Nrf2 signaling pathway to attenuate the ferroptosis of ovarian granulosa cells.

Clinical and Laboratory Characteristics of Infertile Women with Premature Ovarian Insufficiency

Background: Premature ovarian insufficiency (POI) is the preferred term for the condition that was previously referred to as premature menopause or premature ovarian failure. The condition differs from menopause in that there are varying and residual ovarian functions. This study aimed to analyze the clinical characteristics of infertile women with premature ovarian insufficiency. Material & Methods: This prospective study was conducted at the OPD of Impulse Fertility Center, Impulse Hospital, Dhaka, Bangladesh. The study was conducted for a period from January 2022 to December 2022. A total of 23 patients who received outdoor treatment during the study period were purposively selected as sample size. A purposive sampling technique was followed in this study. After approval by the Ethics committee of the Hospital, informed consent from the respondents was obtained after explaining the purpose of the study to them. To be included in the study, respondents should have met the classical definition of infertility defined by the WHO as the inability of a sexually active non-contraceptive using woman to have a live birth after 12 or more months of regular sexual intercourse without a malefactor. Women who had male-factor infertility were excluded. In this study live birth was used as a measure of proven fertility (Because couples desire children, not simply pregnancies, infertility affects couples regardless of whether the etiology lies in conception or the progression of the pregnancy). All the necessary laboratory investigations were done. A questionnaire was developed and data were collected by interviewing the patients and some data were collected from the laboratory results. Data were processed and analyzed by SPSS 19 version. Results: All patients had normal puberty, and menarche occurred at ages 11–15, followed by a regular menstrual cycle. Women who presented with oligomenorrhea were younger than patients with infertility. The mean period of oligomenorrhea before diagnosis was 0.9 years in the oligomenorrhea group and 1.8 years in the infertility group. The mean age when the infrequent periods started was 28 years in the group with oligomenorrhea and 29 years in the group with infertility, and their anthropometric characteristics were not different. In most patients, the FSH levels on day 3 of their menstrual cycle were less than 25?mU/ml. FSH levels >25?mU/ml were confirmed in two patients with oligomenorrhea and five patients in the infertility group. AMH levels were low than 1.0?ng/ml (considered to be in poor ovarian reserve ranges: from 0.13 to 1.0?ng/ml in patients with oligomenorrhea and from 0.13 to 0.9?ng/ml in the infertility group. Transvaginal ultrasound-determined AFC on menstrual cycle days 4–8 was accessed in all patients. We consider AFC?<?6 small follicles (diameters 3–9?mm) as a low ovarian reserve indicator. The lowest AFC (one small follicle) was noted in one infertile patient with oligomenorrhea. Conclusion: This study concluded that the subjects usually present with menstrual irregularity (oligomenorrhea) or infertility, and after proper evaluation, their poor ovarian reserve can be confirmed and an occult form of POI established. women who presented with only oligomenorrhea were younger than infertile patients; therefore, menstrual irregularity may be the earliest clinical symptom of occult POI.

Cardiovascular Risk Factors in Premature Ovarian Insufficiency using Hormonal Therapy / Fatores de risco cardiovascular na insuficiência ovariana prematura em uso de terapia hormonal

Abstract Objective Premature ovarian insufficiency (POI) is characterized by early hypoestrogenism. An increased risk of cardiovascular (CV) disease is a long-term consequence of POI. A challenge of hormone therapy (HT) is to reduce the CV risk. Methods Cross-sectional study with lipid profile analysis (total cholesterol, LDL-C, HDL-C, VLDL-C and triglycerides), blood glucose levels and arterial blood pressure of women with POI using HT, compared with age and BMI-matched women with normal ovarian function (controls). Results The mean age and BMI of 102 POI patients using HT and 102 controls were 37.2 ± 6.0 and 37.3 ± 5.9 years, respectively; 27.0 ± 5.2 and 27.1 ± 5.4 kg/m2. There wasn't difference between groups in arterial systolic and diastolic blood pressure, blood glucose levels, total cholesterol, LDL-C, VLDL-C and triglycerides. HDL-C levels were significantly higher in the POI group (56.3 ± 14.6 and 52 ± 13.9mg/dL; p = 0.03). Arterial hypertension was the most prevalent chronic disease (12% in the POI group, 19% in the control group, p = ns), followed by dyslipidemia (6 and 5%, in POI and control women). Conclusion Women with POI using HT have blood pressure levels, lipid and glycemic profile and prevalence of hypertension and dyslipidemia similar to women of the same age and BMI with preserved gonadal function, in addition to better HDL levels.

Resumo Objetivo A insuficiência ovariana prematura (IOP) é caracterizada pelo hipoestrogenismo precoce. Risco aumentado de doença cardiovascular (CV) é uma consequência a longo prazo da IOP e um desafio da terapia hormonal (TH) é reduzir o risco CV. Métodos Estudo transversal com análise do perfil lipídico (colesterol total, LDL-C, HDL-C, VLDL-C e triglicerídeos), glicemia e pressão arterial de mulheres com IOP em uso de TH, em comparação a mulheres com função ovariana normal (controles) pareadas por idade e IMC. Resultados A média de idade e IMC de 102 pacientes com IOP em uso de TH e 102 controles foi de 37,2 ± 6,0 e 37,3 ± 5,9 anos, respectivamente; 27,0 ± 5,2 e 27,1 ± 5,4 kg/m2. Não houve diferença entre os grupos na pressão arterial sistólica e diastólica, glicemia, colesterol total, LDL-C, VLDL-C e triglicerídeos. Os níveis de HDL-C foram significativamente maiores no grupo IOP (56,3 ± 14,6 e 52 ± 13,9mg/dL; p = 0,03). A hipertensão arterial foi a doença crônica mais prevalente (12% no grupo POI, 19% no grupo controle, p = ns), seguida da dislipidemia (6 e 5%, no grupo POI e controle). Conclusão Mulheres com IOP em uso de TH apresentam níveis pressóricos, perfil lipídico e glicêmico e prevalência de hipertensão e dislipidemia semelhantes às mulheres da mesma idade e IMC com função gonadal preservada, além de melhores níveis de HDL.

Efficacy of Zishen Ankun Granules in Treatment of Mice with Immune Premature Ovarian Insufficiencyon and Its Influence on Cytokines IL-2、IL-6 and TGF-β / 世界科学技术-中医药现代化

Objective To observe efficacy of Zishen Ankun Granules in treatment of mice with immune premature ovarian insufficiency(POI)and its influence on cytokines interleukin 2(IL2),interleukin 6(IL6)and transforming growth factor-β(TGF-β)Methods Screening female mice with regular estrus to establish immune POI mice model with zona pellucida 3(ZP3)as antigen.Low and high doses of Zishen Ankun Granules were set up for treatment,and estradiol valerate was used as a positive control.Observing the estrous cycle of mice by papanicolaou staining,observing the morphological changes of ovarian tissues by HE staining,and detecting serum estradiol(E2),follicle-stimulating hormone(FSH),anti mullerian hormone(AMH),IL2,IL6 and TGF-β levels by ELISA.Results Compared with the blank control group,the estrous cycle of model group was prolonged and disordered,inflammatory cell were infiltrated in the ovarian stroma,growth follicles were decreased,and atresia follicles were increased(P<0.05,P<0.01);serum E2,AMH and TGF-βlevels were decreased(P<0.01),serum FSH,IL2 and IL6 levels were increased(P<0.01).Compared with model group,the estrous cycle disorder and inflammatory cell infiltration were improved in each treatment group,in which the growing follicles increased in the high dose group of Zishen Ankun and the atretic follicles decreased in the estradiol valerate group(P<0.05);serum E2,AMH and TGF-βlevels were increased in Zishen Ankun high dose group and estradiol valerate group(P<0.05,P<0.01),Serum FSH and IL6 levels in Zishen Ankun high-dose group were decreased(P<0.05),Serum IL2 levels was decreased in each treatment group(P<0.05,P<0.01).Conclusion Zishen Ankun granules can improve the levels of serum reproductive hormone,IL2,IL6 and TGF-βin immune POI mice,reduce the inflammatory response of ovary,regulate immune disorders,improve the ovarian reserve capacity,and play a protective role on the ovary.

ffects of Zuogui Pills on kidney-yin deficiency syndrome of premature ovarian insufficiency rats / 国际中医中药杂志

Objective:To study the effects of Zuogui Pills on rats with kidney-yin deficiency syndrome of premature ovarian insufficiency.Methods:Totally 40 SD female unmated rats were randomly divided into blank group, model group, Zuogui Pills group and Bujiale group, with 10 rats in each group. Except for blank group, rats in other groups were subcutaneously injected with ZP3 and gavaged with levothyroxine sodium to induce kidney-yin deficiency syndrome model of premature ovarian insufficiency. At the same time of modeling, Zuogui Pills group and Bujiale group received corresponding drugs for gavage, and the other groups received corresponding solvent for gavage, once a day, for consecutive 21 days. On day 0, 7, 14 and 21, ear temperature and body weight of rats were measured, and the ovarian index, uterus index and thyroid index were calculated. Serum levels of adenosine cyclic phosphate (cAMP), cyclic guanosine phosphate (cGMP), Cortisol, follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E 2) were detected by ELISA. Pathological changes of ovary was observed with HE staining. Results:On day 14 and 21, compared with model group, the body weight of rats in Zuogui Pills group increased ( P<0.05), and the ear temperature decreased ( P<0.05); compared with model group, the ovarian index, uterine index and thyroid index of rats in Zuogui Pills group decreased ( P<0.05), the levels of serum cAMP/cGMP, cortisol, FSH and LH decreased ( P<0.05), and the level of E 2 increased ( P<0.05). Conclusion:Zuogui Pills have certain improvement effect on rats with kidney-yin deficiency syndrome induced by levothyroxine sodium tablets combined with ZP3.

Qirong Tablets inhibits apoptosis of ovarian granulosa cells via PI3K/Akt/ HIF-1 signaling pathway / 中国中药杂志

This study aims to observe the effect and explore the mechanism of Qirong Tablets in the treatment of premature ovarian insufficiency(POI) in mice via the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/hypoxia inducible factor 1(HIF-1) signaling pathway. Sixty SPF female BALB/c mice were randomly divided into normal group, model group, positive control group, Qirong Tablets low-, medium-and high-dose group. The normal group was intraperitoneally injected with the same amount of normal saline, and the other groups were intraperitoneally injected with cyclophosphamide 120 mg·kg~(-1)·d~(-1) once to establish a POI animal model. After the model was successfully established, the low-, medium-and high-dose groups of Qirong Tablets were administered orally with 0.6, 1.2, 2.4 mg·kg~(-1)·d~(-1) respectively. The positive control group was given 0.22 mg·kg~(-1)·d~(-1) Clementine Tablets by intragastric administration, and the normal group and model group were given intragastric administration with the same amount of normal saline, and the treatment was 28 d as a course of treatment. After drug intervention, enzyme-linked immunosorbent assay(ELISA) was employed to measure the levels of estradiol(E_2), follicle-stimulating hormone(FSH), luteinizing hormone(LH), and anti-mullerian hormone(AMH) in peripheral blood, and hematoxylin-eosin(HE) staining to observe the ovarian tissue. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay was used to detect the apoptosis of granulosa cells, and Western blot to determine the expression levels of B-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), caspase-3, PI3K, Akt, and HIF-1. Compared with the normal group, the modeling of POI caused loose or destroyed ovarian tissue with vacuolar structures, edema and fibrosis in the ovarian interstitium, disordered or loose arrangement of granulosa cells, and reduced normal follicles. Compared with the model group, drug interventions restored the ovarian tissue and follicles at all the development stages and reduced atretic follicles. Compared with the normal group, the modeling of POI lowered the serum level of E_2 and AMH(P<0.01), and elevated the level of FSH and LH(P<0.01). Compared with the model group, high-dose Qirong Tablets elevated the levels of E_2 and AMH(P<0.05), and lowered the levels of FSH and LH(P<0.05). Compared with the normal group, the modeling of POI up-regulated the protein levels of PI3K, Akt, HIF-1, Bax, and caspase-3 and down-regulated the protein level of Bcl-2 in the ovarian tissue(P<0.01). Compared with the model group, low-, medium-, and high-dose Qirong Tablets down-regulated the protein levels of PI3K, Akt, HIF-1, Bax, and caspase-3 proteins and up-regulated the protein level of Bcl-2 in the ovarian tissue(P<0.05). In conclusion, Qirong Tablets can up-regulate the expression Bcl-2, down-regulate the expression of Bax and caspase-3 in POI mice. Qirong Tablets may inhibit the apoptosis of follicular granulosa cells in mice, thereby delaying ovarian aging, improving reproductive axis function, and strengthening ovarian reserve capacity, which may be associated with the inhibition of PI3K/Akt/HIF-1 pathway.

Protective Mechanism of Zhuluan Decoction in Rats with Premature Ovarian Insufficiency Based on PI3K/Akt/mTOR Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effect of Zhuluan decoction on the ovarian reserve function of rats with cyclophosphamide-induced premature ovarian insufficiency， and explore the protective mechanism of Zhuluan decoction in the rat model of premature ovarian insufficiency based on the phosphatidylinositol-3-kinases/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway. MethodSixty female SD rats were randomly divided into normal group （n=10） and model group （n=50）. The model group was given intraperitoneal injection of cyclophosphamide （50 mg·kg-1 loading dose on the 1st day+8 mg·kg-1 low-dose maintenance on the 2nd–15th days）. After successfully modeling， the rats were randomly divided into model group， positive drug （progynova） group （0.1 mg·kg-1·d-1）， and low-， medium-， and high-dose Zhuluan decoction groups （14， 28， 56 g·kg-1·d-1 ）， with 10 rats in each group. The model group and the normal group were given equal volume of distilled water by gavage， once a day， continuous administration for 21 d. The estrous cycle and body weight of rats in each group were detected， and the ovarian organ index and uterine organ index were calculated. The ovarian tissue pathology and ovarian follicle counts at all levels were determined by hematoxylin-eosin （HE） staining. The content of the serum antimullerian hormone （AMH）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and inhibin-B （INH-B） of rats was determined by enzyme-linked immunosorbent assay （ELISA）， and the protein expression levels of PI3K， Akt， mTOR in the rat ovarian tissue were determined by Western blot. The microtubule-associated protein 1 light chain 3B （LC3B） protein expression in the rat ovarian tissue was determined by immunohistochemistry. ResultAs compared with the blank group， the estrous cycle of rats in the model group was disordered， the body weight， ovarian organ index， and uterine organ index decreased， the number of primordial follicles decreased， and the number of secondary follicles and atretic follicles increased. In the model group， FSH increased （P<0.01）， LH increased （P<0.05）， AMH level decreased （P<0.05）， the protein expression levels of PI3K， Akt， and mTOR in the ovarian tissue decreased （P<0.01）， and the protein expression level of LC3B increased significantly （P<0.01）. As compared with the model group， the above indexes were improved in the progynova group and different doses of Zhuluan decoction groups， the content of AMH increased （P<0.05）， and FSH decreased （P<0.05）. In the progynova group and different doses of Zhuluan decoction groups， the protein expression level of LC3B decreased obviously （P<0.01）， and the protein expression levels of PI3K， Akt， and mTOR all showed an increasing trend. Moreover， there was a statistically significant difference in the progynova group and low- and medium-dose Zhuluan decoction groups （P<0.05）. ConclusionZhuluan decoction may inhibit the occurrence of excessive autophagy in ovarian granulosa cells by activating the PI3K/Akt/mTOR pathway， thereby reversing the effect of modeling on ovarian reserve in rats.

Animal Model of Premature Ovarian Insufficiency Based on Data Mining / 中国实验方剂学杂志

ObjectiveTo study the modeling characteristics of the animal model of premature ovarian insufficiency and provide references for the standardization of the animal model of premature ovarian insufficiency， thus offering a better research basis to the pathogenesis， diagnosis， and treatment of the disease. MethodThe animal experimental literature of premature ovarian insufficiency in the past decade was obtained by searching China National Knowledge Infrastructure （CNKI）， Wanfang， VIP Chinese Technology Periodical Database， China Biology Medicine （CBM）， and PubMed. The types of experimental animals， modeling methods， dosage， administration scheme， modeling standards， and detection indicators were summarized. The frequency analysis was performed with Excel， the association rule analysis was performed with SPSS Modeler 18.0， and the results were visually upgraded with Cytascape 3.6.1. ResultA total of 281 articles were included， and most animal experiments on premature ovarian insufficiency were performed on SD rats or BALB/c mice. Most modeling methods were iatrogenic induction， and the main modeling drug was cyclophosphamide. Erestrous cycle disorder was selected as the modeling standard. Ovarian histomorphology， estradiol， follicle stimulating hormone， and estrous cycle were selected as the detection indicators to comprehensively evaluate the model from multiple aspects. ConclusionSD rats are mostly used to induce modeling in animal experiments on premature ovarian insufficiency through first intraperitoneal injection of cyclophosphamide 50 mg·kg-1 and 8 mg·kg-1·d-1 from the next day for 14 d， which has the advantages of easy operation， high modeling rate， and consistency with the disease pathogenesis. This paper can provide references for basic animal experimental research of premature ovarian insufficiency.

Effect and Mechanism of Nongsuo Dangguiwan in Relieving Oxidative Stress in Premature Ovarian Insufficiency Rats Based on Nrf2/HO-1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect and mechanism of Nongsuo Dangguiwan in improving ovarian oxidative stress in rats with ovarian dysfunction. MethodThirty-six adult female SD rats were randomly divided into normal group， model group， positive drug group （Femoston， 0.3 mg·kg-1）， and high-， medium-， and low-dose groups of concentrated Nongsuo Dangguiwan （2.08， 4.16， 8.32 g·kg-1）， with six rats in each group. Rats， except for those in the normal group， were injected with 80 mg·kg-1 vinyl cyclohexene dioxide （VCD） per day for 14 consecutive days to induce ovarian dysfunction. From the 15th day， rats were treated with corresponding drugs by gavage， while those in the model group received 2 mL·kg-1 saline， once daily for 28 consecutive days. The ovarian index， levels of related hormones including estradiol （E2）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and anti-mullerian hormone （AMH） in serum， as well as superoxide dismutase （SOD） activity and glutathione （GSH） content in serum were measured by enzyme-linked immunosorbent assay （ELISA）. The malondialdehyde （MDA） content in serum was detected by the thiobarbituric acid （TAB） method. Ovarian morphology was observed by hematoxylin-eosin （HE） staining. The expression of nuclear factor E2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， SOD2， and SOD1 in ovarian tissues was detected by immunohistochemistry （IHC） and Western blot. ResultCompared with the normal group， the model group showed a significant reduction in growing follicles in the ovary， loose arrangement of granulosa cells in the follicle， decreased body weight， ovarian index， and serum AMH and E2 levels， increased LH and FSH levels （P<0.01）， reduced levels of SOD and GSH in serum （P<0.01）， and increased MDA level （P<0.01）. Compared with the model group， the groups with drug intervention showed increased ovarian index （P<0.05， P<0.01）， increased serum E2 level （P<0.05， P<0.01）， decreased FSH， AMH， and LH levels （P<0.05， P<0.01）， increased number of growing follicles in the ovary， potentiated SOD activity in serum， increased GSH content， decreased MDA content （P<0.05， P<0.01）， and up-regulated expression levels of Nrf2， HO-1， SOD2， and SOD1 proteins in ovarian tissues （P<0.05， P<0.01）. ConclusionNongsuo Dangguiwan can regulate serum hormone levels， increase the expression of Nrf2， HO-1， SOD2， and SOD1 in ovarian tissues， and improve ovarian antioxidant capacity to resist oxidative stress injury， thereby improving ovarian reserve function.

Effect of acupuncture at "Zhibian" (BL 54) through "Shuidao" (ST 28) on the expression of apoptosis-related factors in rats with premature ovarian insufficiency based on oxidative stress / 中国针灸

OBJECTIVE@#To explore the possible mechanism of acupuncture at "Zhibian" (BL 54) through "Shuidao" (ST 28) on premature ovarian insufficiency (POI) from the perspective of oxidative stress.@*METHODS@#Sixty female SD rats were randomly divided into a blank group, a model group, a sham acupuncture group, a medication group, and an acupuncture group, 12 rats in each group. Except the blank group, the rats in the remaining groups were intraperitoneally injected with cyclophosphamide to establish the POI model. After the model was successfully established, the rats in the acupuncture group were treated with acupuncture at "Zhibian" (BL 54) through "Shuidao" (ST 28), with a depth of about 12 mm, and the needle was retained for 30 min; the acupuncture was given once a day, for a total of 4 weeks. The rats in the sham acupuncture group were treated with blunt-head needle to tap the skin surface of "Zhibian" (BL 54), without penetrating the skin, once a day for 4 weeks. The rats in the medication group were treated with estradiol valerate by gastric gavage for 4 weeks. After the intervention, the level of reactive oxygen species (ROS) in the ovarian tissue was detected by fluorescence probe; the expression of c-Jun N-terminal kinase (JNK), forkhead box O1 (FoxO1), tumor suppressor gene protein 53 (p53) and p53 up-regulated modulator of apoptosis (Puma) mRNA and protein in ovarian tissue were detected by real-time fluorescence quantitative PCR and Western blot.@*RESULTS@#Compared with the blank group, the level of ROS and the expression of JNK mRNA, p-JNK protein, FoxO1, p53, Puma mRNA and protein in the ovarian tissue in the model group were increased (P<0.01). Compared with the model group, the level of ROS and the expression of p-JNK protein, FoxO1, p53, Puma mRNA and protein in the ovarian tissue in the sham acupuncture group were slightly reduced, but the difference was not statistically significant (P>0.05). The level of ROS and the expression of JNK mRNA, p-JNK protein, FoxO1, p53, Puma mRNA and protein in the ovarian tissue in the acupuncture group and the medication group were reduced (P<0.01).@*CONCLUSION@#Acupuncture at "Zhibian" (BL 54) through "Shuidao" (ST 28) could improve the level of oxidative stress, down-regulate the expression of apoptosis-related factors JNK, FoxO1, p53 and Puma induced by oxidative stress, and inhibit the premature failure of ovarian reserve function caused by apoptosis of ovarian granulosa cells in POI rats.

Fecal incontinence or pelvic organ prolapse among women with premature ovarian insufficiency

Objectives: To assess the status of the pelvic floor muscle (PFM) of premature ovarian insufficiency women (POI women) and the incidence of fecal incontinence (FI) and pelvic organ prolapse (POP). Methods: A secondary analysis of a cross-sectional study with 150 women with POI was performed. Pelvic floor muscle assessment was performed with the PERFECT scale. The subscales POPDI-6 and CRADI-8 of the questionnaire Pelvic Floor Distress Inventory-20 (PFDI-20) were used for pelvic floor symptoms focused on FI and POP. Moreover, FI and POP were also assessed as dichotomous variables (yes/no). Results: Women with FI and POP did not present differences in the PFM assessment across P (p = 0.61), E (p = 0.78), R (p = 0.22), and F (p = 0.79) variables when compared with women with POI; no differences were also seen between women with and without POP according the pelvic muscles: P (p = 0.91), E (p = 0.99), R (p = 0.62), and F (p = 0.10). Women with FI and POP presented higher scores in all PFDI-20 subscales and total score when compared with the control group (p < 0.05). Conclusions Pelvic floor muscle assessment within POI women with or without FI or POP did not differ. However, PF symptoms are more severe in the FI or POP groups. (AU)

Effect of regulating menstruation and promoting pregnancy acupuncture therapy on negative emotion in patients with premature ovarian insufficiency / 中国针灸

OBJECTIVE@#To observe the influence of regulating menstruation and promoting pregnancy acupuncture therapy on negative emotions in patients with premature ovarian insufficiency (POI).@*METHODS@#A total of 60 patients with POI were randomly divided into an acupuncture group and a western medication group, 30 cases in each group. The acupuncture group was treated with regulating menstruation and promoting pregnancy acupuncture therapy at Baihui (GV 20), Shenting (GV 24), Guanyuan (CV 4), Sanyinjiao (SP 6), Shenshu (BL 23), Ciliao (BL 32), etc. once a day, 5 times a week for 3 months. The western medication group was treated by oral administration of climen. The drug was given 1 tablet a day for 21 days and was stopped for 1 week as a course. The treatment was required 3 consecutive courses. The self-rating anxiety scale (SAS) score, modified Kupperman index (KI) score, agitated and depressive symptom scores in KI and serum level of follicle stimulating hormone (FSH) before and after treatment were compared between the two groups.@*RESULTS@#After treatment, the SAS scores, KI scores and serum levels of FSH in the two groups and the scores of agitated and depressive symptom in the acupuncture group were lower than those before treatment (@*CONCLUSION@#Regulating menstruation and promoting pregnancy acupuncture therapy can effectively improve the negative emotions of patients with POI and reduce serum level of FSH .

Genetic study on a consanguineous Chinese family with premature ovarian insufficiency caused by a missense mutation of PSMC3IP gene / 中国医师杂志

Objective:To explore the genetic etiology for a premature ovarian insufficiency (POI) patient from a consanguineous Chinese family, and to provide basis for genetic counseling and fertility counseling.Methods:Whole-exome sequencing was performed using DNA extracted from the blood sample of POI patient. Suspected pathogenic mutation was analyzed by bioinformatics methods and verified by Sanger sequencing. The pathogenicity of the variation was assessed according to the ACMG genetic variation classification criteria and guidelines.Results:A homozygous variation, c. 32G>T (p.G11V), of PSMC3IP was identified in the patient. Bioinformatics analysis revealed that the variation was conserved in different animal species, and this variation was classified as possible pathogenic variation according to the ACMG genetic variation classification criteria and guidelines.Conclusions:The homozygous missense variation of PSMC3IP is the cause of the POI patient in this family. We are reporting for the first time the missense variation in PSMC3IP gene caused POI, which enriched the mutation spectrum of PSMC3IP and provided the basis for genetic counseling and fertility guidance of this family.

Acupuncture-moxibustion treatment by stages based on the theory of "transformation of / 中国针灸

OBJECTIVE@#To compare the efficacy between acupuncture-moxibustion treatment by stages and femoston for premature ovarian insufficiency (POI).@*METHODS@#A total of 66 patients with POI were randomly divided into an observation group (33 cases, 3 cases dropped off) and a control group (33 cases, 2 cases dropped off). The patients in the observation group, based on the theory of "transformation of @*RESULTS@#Compared before treatment, the serum levels of FSH and LH were decreased (@*CONCLUSION@#Acupuncture- moxibustion treatment by stages based on the theory of "transformation of

Effect of Huluan Decoction on SIRT1/NF-κB/p53/p21 Pathway in Model Mice with Premature Ovarian Insufficiency / 中国实验方剂学杂志

Objective:To observe the effects of Huluan decoction on the expression of apoptosis-related protein silence information regulator 1 （SIRT1）， tumor suppressor gene （p53）， acetylated p53 （Ac-p53）， cyclin-dependent kinase inhibitor （p21）， and nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） p65 and changes in ovarian tissue structure of a mouse model with premature ovarian insufficiency （POI） induced by tripterygium glycosides. Method:Fifty female C57BL/6J mice were randomly divided into the blank group， model group， low- and high-dose Huluan decoction groups， and western medicine （estradiol valerate） group. After intragastric administration of tripterygium glycosides at 80 mg·kg^-1·d^-1 for 14 successive days， mice in the low- and high-dose Huluan decoction groups and western medicine group were treated with Huluan decoction at 1.6 g·kg^-1·d^-1 and 6.2 g·kg^-1·d^-1 and estradiol valerate at 0.13 mg·kg^-1·d^-1， respectively， by gavage since the 15^th day， while those in the blank group were provided with an equal amount of distilled water for 21 consecutive days. Following the last administration， the blood was sampled for detecting the levels of serum anti-Müllerian hormone （AMH）， follicle-stimulating hormone （FSH）， and luteinizing hormone （LH） by ELISA and the ovary was harvested for observing the morphological changes by HE staining. The mRNA expression levels of p53 and p21 were measured by Real-time polymerase chain reaction （Real-time PCR）， and the protein expression levels of SIRT1， p53， p21， Ac-p53， NF-<italic>κ</italic>B were assayed by Western blot. Result:Compared with the blank group， the model group exhibited a disordered estrous cycle， diminished ovarian volume， decreased number of follicles at various developmental stages， reduced AMH， and elevated FSH and LH， and elevated p53，p21，Ac-p53，NF-<italic>κ</italic>B protein expression and p53，p21 mRNA， reduced SIRT1 protein expression（<italic>P</italic><0.01）. As revealed by the comparison with the model group， each medication group displayed an increased number of follicles， elevated AMH， reduced FSH and LH （<italic>P</italic><0.01）， up-regulated SIRT1 protein expression （<italic>P</italic><0.01）， and significantly down-regulated mRNA and protein expression of aging-related genes p53， p21， and down-regulated Ac-p53，NF-<italic>κ</italic>B protein expression （<italic>P</italic><0.01）. Conclusion:Huluan decoction significantly reverses the aging process and improves ovarian function possibly by boosting the activity of SIRT1/NF-<italic>κ</italic>B/p53/p21 pathway in ovarian cells， changing the apoptotic state， increasing the growing and mature follicles， and reducing the atretic follicles.

Mulheres submetidas ao transplante de medula óssea: impacto dos quimioterápicos e da doença do enxerto contra o hospedeiro sobre o sistema reprodutor / Women undergoing bone marrow transplantation: impact of chemotherapy and graft-versus-host disease (GVHD) on the feminine reproductive system

O transplante de medula óssea (TMO) é um procedimento indicado para o tratamento de doenças hematológicas, que afetam muitas mulheres jovens. O aperfeiçoamento dos cuidados durante o TMO proporciona altos índices de cura e de sobrevida. No entanto, pode deixar sequelas em vários órgãos e sistemas, entre eles o sistema reprodutor e os órgãos genitais, impactando negativamente a qualidade de vida das receptoras do TMO. O objetivo desta publicação foi realizar uma revisão narrativa sobre o tema e propor um protocolo assistencial que torne acessível os cuidados relacionados à saúde sexual e reprodutiva a esse grupo especial de mulheres, baseado em dados clínicos de um ambulatório de assistência ginecológica às mulheres transplantadas no Hospital Amaral Carvalho, em Jaú, no interior de São Paulo.(AU)

Bone marrow transplantation (BMT) is indicated for the treatment of hematological diseases which affect many young women. The improvement of care during BMT procedures provides higher cure and survival rates. however, it can cause sequelae in various organs and systems, including the reproductive system and genitals, negatively impacting quality of life. The purpose of this publication is to present a narrative review related to this theme and to propose a healthcare protocol that allows sexual and reproductive care in this special group of patients, based on the clinical experience of a gynecological outpatient clinic at the Amaral Carvalho Hospital, in Jaú (SP) which specifically care for these women.(AU)