Estrogen Levels Research Articles

The effect of Mongolian Medicine Chagan Gaoyou-4 Powder on bone mineral density induced by retinoic acid in rats

Objective: To investigate the effects of Chagan-Gaoyou-4 powder on spine bone mineral density and serum estrogen and estrogen receptor alpha (ERa) and estrogen receptor beta (ERβ) protein expression in bone tissue induced by retinoic acid in rats. Methods: Sixty 3-month-old SPF female SD rats were randomly divided into model group (retinoin group), normal group (SHAM group), Mongolian medicine group (Chagan-Gaoyou 4 group), and control group (Gushukang granule group). After 14 days of administration, spinal Bone Mineral Density (BMD) was measured, and external manifestations were observed. Results: Compared with the SHAM group, the BMD values of all medication groups decreased (P < 0.05, P < 0.001). Compared with the model group, the spinal BMD value of the Chagan Gaoyou-4 group was increased after 21 days of treatment, and the difference was statistically significant (P < 0.01, P < 0.05). After 21 days, the spinal BMD value of all treatment groups was increased (P < 0.05), the spinal BMD value was also increased, especially Chagan Gaoyou-4 group was more obviously increased (P < 0.05). Compared with the normal group, ERa and ERβ levels in the model group were significantly decreased (P < 0.001), and ERa and ERβ levels in all drug groups were significantly increased (P < 0.001) compared with the model group. After 21 days of treatment, there was no significant difference among the three treatment groups (P > 0.05). ERa and ERβ levels in the Chagan Gaoyou-4 group were higher than those in the model group (P < 0.05), but there was no significant difference between the group and the Gushukang granule group (P > 0.05). The E2 content of the Chagan Gaoyou-4 group was higher than that of the model group (P < 0.05). The mRNA expression of estrogen receptor ERa and ERβ in bone tissue of the left proximal femur was detected by RT-PCR. Compared with the model group, the expression of ERa and ERβ mRNA in the Chagan Gaoyou-4 group and Gushukang granule group was increased. Compared with the model group, ERp mRNA expression was increased in the Chagan Gaoyou-4 powder medium-dose group. Compared with the Gushukang granule group, there was no significant difference in ERm RNA expression in the Chagan Gaoyou-4 dose groups. Chagan Gaoyou-4 can up-regulate the expression of ERa and ERβ induced by retinoic acid in rats, indicating that the Chagan Gaoyou-4 powder group may promote bone formation, regulate bone resorption, improve bone mineral density, and achieve the purpose of preventing and treating OP by increasing estrogen level, stimulating estrogen receptor, and increasing the expression of ERa and ERβ in bone tissue. Conclusion: Chagan-Gaoyu-4 powder may have estrogen-like effects on the bone tissue of rats induced by retinoic acid and may increase the level of serum estrogen, ERa, and ERβ protein expression, thereby improving the spinal BMD of experimental rats. The Chagan-Gaoyou-4 powder group could improve the general condition of osteoporosis induced by retinoic acid in rats.

The intersection between menopause and depression: overview of research using animal models.

Menopausal women may experience symptoms of depression, sometimes even progressing clinical depression requiring treatment to improve quality of life. While varying levels of estrogen in perimenopause may contribute to an increased biological vulnerability to mood disturbances, the effectiveness of estrogen replacement therapy (ERT) in the relief of depressive symptoms remains controversial. Menopausal depression has a complex, multifactorial etiology, that has limited the identification of optimal treatment strategies for the management of this psychiatric complaint. Nevertheless, clinical evidence increasingly supports the notion that estrogen exerts neuroprotective effects on brain structures related to mood regulation. Indeed, research using preclinical animal models continues to improve our understanding of menopause and the effectiveness of ERT and other substances at treating depression-like behaviors. However, questions regarding the efficacy of ERT in perimenopause have been raised. These questions may be answered by further investigation using specific animal models of reduced ovarian function. This review compares and discusses the advantages and pitfalls of different models emulating the menopausal stages and their relationship with the onset of depressive-like signs, as well as the efficacy and mechanisms of conventional and novel ERTs in treating depressive-like behavior. Ovariectomized young rats, middle-to-old aged intact rats, and females treated with reprotoxics have all been used as models of menopause, with stages ranging from surgical menopause to perimenopause. Additionally, this manuscript discusses the impact of organistic and therapeutic variables that may improve or reduce the antidepressant response of females to ERT. Findings from these models have revealed the complexity of the dynamic changes occurring in brain function during menopausal transition, reinforcing the idea that the best approach is timely intervention considering the opportunity window, in addition to the careful selection of treatment according to the presence or absence of reproductive tissue. Additionally, data from animal models has yielded evidence to support new promising estrogens that could be considered as ERTs with antidepressant properties and actions in endocrine situations in which traditional ERTs are not effective.

Antipsychotic-induced prolactin elevation in premenopausal women with schizophrenia: associations with estrogen, disease severity and cognition.

Antipsychotic-induced prolactin elevation may impede protective effects of estrogens in women with schizophrenia-spectrum disorders (SSD). Our study sought to confirm whether the use of prolactin-raising antipsychotics is associated with lower estrogen levels, and to investigate how estrogen and prolactin levels relate to symptom severity and cognition in premenopausal women with SSD. This cross-sectional study included 79 premenopausal women, divided in three groups of women with SSD treated with prolactin-sparing antipsychotics (n = 21) or prolactin-raising antipsychotics (n = 27), and age-matched women without SSD (n = 31). Circulating 17β-estradiol was compared among groups. In patients, we assessed the relationship between prolactin and 17β-estradiol, and the relationships of these hormones to symptom severity and cognition, using correlation analyses and backward regression models. In women receiving prolactin-raising antipsychotics, 17β-estradiol levels were lower as compared to both other groups (H(2) = 8.34; p = 0.015), and prolactin was inversely correlated with 17β-estradiol (r=-0.42, p = 0.030). In the prolactin-raising group, 17β-estradiol correlated positively with verbal fluency (r = 0.52, p = 0.009), and 17β-estradiol and prolactin together explained 29% of the variation in processing speed (β17β-estradiol = 0.24, βprolactin=-0.45, F(2,25) = 5.98, p = 0.008). In the prolactin-sparing group, 17β-estradiol correlated negatively with depression/anxiety (r=-0.57, p = 0.014), and together with prolactin explained 26% of the variation in total symptoms (β17β-estradiol=-0.41, βprolactin = 0.32, F(2,18) = 4.44, p = 0.027). In women with SSD, antipsychotic-induced prolactin elevation was related to lower estrogen levels. Further, estrogens negatively correlated with symptom severity and positively with cognition, whereas prolactin levels correlated negatively with cognition. Our findings stress the clinical importance of maintaining healthy levels of prolactin and estrogens in women with SSD.

Induction of insulin resistance in female mice due to prolonged phenanthrene exposure: Unveiling the low-dose effect and potential mechanisms

Phenanthrene (Phe) is a commonly occurring polycyclic aromatic hydrocarbon (PAH) found in various food sources and drinking water. Previous studies have shown that long-term exposure to Phe in male mice leads to insulin resistance in a dose-dependent manner. However, the effect of Phe on glucose homeostasis in female mice remains unknown. To address this knowledge gap, female Kunming mice were exposed to Phe through their drinking water at concentrations of 0.05, 0.5, and 5 ng/mL. After 270 d of exposure, we surprisingly discovered a low-dose effect of Phe on insulin resistance in female mice, which differed from the effect observed in male mice and showed sexual dimorphism. Specifically, insulin resistance was only observed in the 0.05 ng/mL treatment, and this low-dose effect was also reflected in the concentration of Phe in white adipose tissue (WAT). Differences in metabolic enzyme activities in the liver may potentially explain this effect. The observed sexual dimorphism in Phe exposure could be attributed to variations in estrogen (E2) level and estrogen receptor beta (ERβ) expression in WAT. These findings highlight the association between environmental factors and the development of insulin resistance, emphasizing the pathogenic effect of even low doses of Phe. Moreover, sex dependent-effect should be given more attention when studying the toxic effects of environmental pollutants.

Oral dehydroepiandrosterone supplementation enhances osteoporotic fracture healing in the OVX rats

Osteoporosis easily causes delayed fracture union, even non-union. It has been demonstrated that dehydroepiandrosterone (DHEA) supplementation can increase estrogen levels and improve bone mineral density (BMD) in the elderly, while the role of DHEA on fracture healing remains unknown. This study aimed to elucidate the impact of DHEA supplementation on osteoporotic fracture healing. Seventy-two female Sprague-Dawley rats were used. Forty-eight rats received ovariectomy (OVX), and the remaining rats received a sham OVX operation (sham group). A right transverse femoral osteotomy was performed in all rats at 12 weeks post-OVX. OVX rats were randomly allocated into 2 groups (n = 24 in each group): (i) ovariectomized rats (control group) and (ii) ovariectomized rats treated with DHEA (DHEA group, 5 mg/kg/day). The DHEA supplementation was initiated on the first day post-fracture for 3, 6, and 12 weeks. Fracture healing was evaluated by radiography, histology, biomechanical analysis, and dual-energy X-ray absorptiometry (DEXA). Serum biomarkers were analyzed using enzyme-linked immunosorbent assay (ELISA). At 3 and 6 weeks, radiographs revealed reduced calluses formation and lower radiographic scores in the control group than in other groups. The sham and DHEA groups showed higher BMD and bone mineral content (BMC) at the fracture site than the control group after fracture. Histological analysis revealed the fracture callus was remodeled better in the sham and DHEA groups than in the control group. At the early phase of healing, DHEA supplementation increased osteoblast number, callus area, and cartilage area than the control group. An increased bone area was observed in the DHEA group than in the control group at the late phase of healing. Additionally, improved biomechanical characteristics were observed in both the sham and DHEA groups than those in the control group post-fracture. ELISA showed higher levels of insulin-like growth factor-1 (IGF-1) and 17β-estradiol (E2) in the DHEA group than in the control group post-fracture. Furthermore, the DHEA group exhibited significantly elevated alkaline phosphatase (ALP) and osteocalcin (OC) levels compared to the control group at 6 and 12 weeks. The DHEA group and the control group did not exhibit a notable difference in TRAP-5b levels. The present study demonstrated that the DHEA treatment has a favorable impact on osteoporotic fracture healing by enhancing callus formation, consolidation, and strength in the OVX rats.

Correlation between serum estrogen level and endometrial histology in cases of fibroid uterus in peri-menopausal peroid

Correlation between serum estrogen level and endometrial histology in cases of fibroid uterus in peri-menopausal peroid

Associations of endogenous estrogens, plasma Alzheimer's disease biomarkers, and APOE4 carrier status on regional brain volumes in postmenopausal women.

Women carrying the APOE4 allele are at greater risk of developing Alzheimer's disease (AD) from ages 65-75 years compared to men. To better understand the elevated risk conferred by APOE4 carrier status among midlife women, we investigated the separate and interactive associations of endogenous estrogens, plasma AD biomarkers, and APOE4 carrier status on regional brain volumes in a sample of late midlife postmenopausal women. Participants were enrolled in MsBrain, a cohort study of postmenopausal women (n = 171, mean age = 59.4 years, mean MoCA score = 26.9; race = 83.2% white, APOE4 carriers = 40). Serum estrone (E1) and estradiol (E2) levels were assessed using liquid chromatography-tandem mass spectrometry. APOE genotype was determined using TaqMan SNP genotyping assays. Plasma AD biomarkers were measured using single molecule array technology. Cortical volume was measured and segmented by FreeSurfer software using individual T1w MPRAGE images. Multiple linear regression models were conducted to determine whether separate and interactive associations between endogenous estrogen levels, plasma AD biomarkers (Aβ42/Aβ40, Aβ42/p-tau181), and APOE4 carrier status predict regional brain volume (21 regions per hemisphere, selected a priori); and, whether significant interactive associations between estrogens and AD biomarkers on brain volume differed by APOE4 carrier status. There was no main effect of APOE4 carrier status on regional brain volumes, endogenous estrogen levels, or plasma AD biomarkers. Estrogens did not associate with regional brain volumes, except for positive associations with left caudal middle frontal gyrus and fusiform volumes. The interactive association of estrogens and APOE4 carrier status on brain volume was not significant for any region. The interactive association of estrogens and plasma AD biomarkers predicted brain volume of several regions. Higher E1 and E2 were more strongly associated with greater regional brain volumes among women with a poorer AD biomarker profile (lower Aβ42/40, lower Aβ42/p-tau181 ratios). In APOE4-stratified analyses, these interactions were driven by non-APOE4 carriers. We demonstrate that the brain volumes of postmenopausal women with poorer AD biomarker profiles benefit most from higher endogenous estrogen levels. These findings are driven by non-APOE4 carriers, suggesting that APOE4 carriers may be insensitive to the favorable effects of estrogens on brain volume in the postmenopause.

Case Report: The Effect of Ajwa Date Consumption on Total Cholesterol Levels in Perimenopausal Women

Based on the 2016 Riskesdas, the prevalence of high cholesterol in Indonesia among women aged 35-59 years was 52.9%, and among those aged ≥60 years, it was 58.7%. According to provincial data, the highest percentage of patients with high cholesterol levels at Posbindu and FKTP in Indonesia was in West Papua Province, reaching up to 70%. The concentration of cholesterol levels in women's blood tends to increase with age, particularly for those over 40 years old, due to hormonal factors, especially the decrease in estrogen production and function. The decline in estrogen levels leads to an increase in lipids or total cholesterol, resulting in changes in body fat composition associated with hypercholesterolemia. This case report aims to investigate the impact of consuming Ajwa dates on the total cholesterol levels of perimenopausal women. Results: Ajwa dates contain higher polyphenol concentrations with an impressive nutritional profile that enhances blood lipoproteins. Additionally, dates can increase estrogen levels due to the estradiol and estrone present in their pollen. Several experimental studies have shown a significant difference in total cholesterol levels in subjects before and after consuming Ajwa dates. Conclusion: There is a notable difference in total cholesterol levels before and after the administration of Ajwa dates.

Therapeutic effect of Momordica charantia on cardiomyopathy in a diabetic maternal rat model.

Myocardial structural and functional abnormalities are hallmarks of diabetic cardiomyopathy (DCM), a chronic consequence of diabetes mellitus (DM). Maternal DM affects and increases the risk of heart defects in diabetic mothers compared with nondiabetic mothers. Momordica charantia exhibits antidiabetic effects due to various bioactive compounds that are phytochemicals, a broad group that includes phenolic compounds, alkaloids, proteins, steroids, inorganic compounds, and lipids. Pregnant maternal rats were split into four groups: control (C), M. charantia-treated (MC), type 2 diabetes mellitus (T2DM) (DM), and diabetic (MC + DM) groups. Diabetes mothers had increased serum glucose, insulin, total cholesterol, triglyceride, and low-density lipoprotein cholesterollevels and reduced high-density lipoprotein cholesterollevels. Cardiac biomarkers such as cardiac troponin T (cTnT), creatine kinase-myocardial band (CK-MB), and lactate dehydrogenasewere increased. Hormone levels of follicle-stimulating hormone, luteinizing hormone, progesterone, and estrogen decreased significantly. Inflammatory markers such as interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-α), and vascular adhesion molecule-1 (VCAM-1) were elevated in diabetic mothers. Oxidative stress markers indicated increased malondialdehydeand nitric oxidelevels, while antioxidants such as glutathione, superoxide dismutase, and catalasewere decreased in maternal heart tissue. The levels of apoptotic markers such as tumor suppressor 53 (P53) and cysteine aspartic protease-3 (caspase-3) were significantly greater in diabetic maternal heart tissue. Histopathological analysis revealed heart tissue abnormalities in diabetic maternal rats. M. charantia extract improved maternal diabetes-induced changes in inflammation, antioxidant levels, and heart tissue structure.

P-367 Letrozole or Tamoxifen co-administration during fertility preservation of breast cancer patients: a retrospective cohort study

Abstract Study question How do fertility preservation (FP) outcomes differ between breast cancer patients stimulated with FSH and Tamoxifen versus FSH and Letrozole? Summary answer Letrozole and Tamoxifen co-administration yielded similar number of retrieved and cryopreserved oocytes/embryos, as well as similar oocyte maturation and fertilization and rates. What is known already Oocyte/embryo cryopreservation is recommended before initiation of chemotherapy in cancer patients. The co-administration of Tamoxifen, a selective estrogen receptor modulator, or Letrozole, an aromatase inhibitor, during ovarian stimulation with gonadotropins, are used to limit the potentially harmful effects of a supra-physiological rise in estrogen levels on hormone-sensitive cancers. However, the optimal choice between the two is still unclear and with no sound evidence. Study design, size, duration Retrospective cohort study of all FP treatments of newly diagnosed breast cancer patients in two tertiary centers between 2000-2022. Collected data included 354 FP cycles of 290 patients and compared treatment parameters and outcomes of Tamoxifen or Letrozole co-administration. Participants/materials, setting, methods Breast cancer patients who underwent FP before chemotherapy were included. Patients with a history of radiation or gonadotoxic treatment, as well as those undergoing other fertility preservation treatments, such as ovarian tissue cryopreservation were excluded. Main results and the role of chance Overall, 215 Letrozole and 139 Tamoxifen treatment cycles were compared, with no significant demographic differences. Treatments with Letrozole had higher FSH dosages (3970IU vs. 2839IU, p < 0.001) but shorter duration (9.7 vs. 10.5 days, p < 0.001). The Letrozole group achieved lower peak E2 levels and endometrial thickness (1508pmol/L, 8.1mm vs. 8760pmol/L, 9.5mm, p < 0.001). The mean number of retrieved oocytes was comparable between the groups (13.6 with Letrozole vs 14.7 with Tamoxifen, p = 0.06). In 122 Letrozole and 46 Tamoxifen patients that underwent strictly oocytes cryopreservation the mean number of cryopreserved oocytes was comparable (11 vs. 12.1, p = 0.21). Moreover, in embryo cryopreservation cycles of 71 patients treated with Letrozole compared to 76 patients with Tamoxifen, no difference was observed in the number of cryopreserved embryos (6.2 vs. 7.0, p = 0.12). Oocyte maturation rate among strictly oocyte preservation cycles was comparable (86% with Letrozole vs. 83% with Tamoxifen, p = 0.80), as were fertilization rates in embryo cryopreservation cycles (62% vs. 63%, p = 0.8 and 71% vs. 76%, p = 0.3 in IVF and ICSI with Letrozole and Tamoxifen, respectively). Multivariate analysis, controlling for potential confounders showed no significant correlations between Tamoxifen or Letrozole co-administration and the number of retrieved oocytes or oocyte maturation rate. Limitations, reasons for caution The primary limitation of our study is its retrospective design and the variation in treatment duration and gonadotrophin dosages among groups. Wider implications of the findings This study provides reassurance that both Tamoxifen and Letrozole are equally effective options when co-administered in FP treatments of breast cancer patients. Trial registration number not applicable

Features of intestinal microbiota taxonomic composition and their relation with hormonal and immune status assessed in women with external genital endometriosis

Aim: systemically assessed characteristics of intestinal microbiota taxonomic composition in relation to parameters of hormonal and immune status in patients with external genital endometriosis (EGE).Materials and Methods. The controlled cross-sectional study included 33 patients with ЕGE comprising main group, and 30 healthy women enrolled to control group. All women underwent assessment of hormonal status and cytokine expression levels in peripheral blood. Level of blood hormones estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) was carried out using enzyme-linked immunosorbent and chemiluminescent assays. Expression levels of cytokines such as interleukin (IL) IL-6, IL-8, tumor necrosis factor alpha (TNF-α) were analyzed by an enzyme-linked immunosorbent assay. Taxonomic composition of intestinal microbiota at the level of phyla and genera was carried out by 16S ribosomal RNA genesequencing. Microbial community α-diversity, the Chao1, ACE, and Sobs indices were used.Results. The concentration of blood E2 in main group was significantly higher compared to control group. Also, women with ЕGE had higher plasma concentrations for IL-6, IL-8, IL-17 and TNF-α compared to those in control group (p < 0.001). While analyzing bacterial community α-diversity in main group, Chao1 index was found to be significantly decreased. At the phylum level, the Firmicutes/Bacteroidetes ratio was increased in patients with ЕGE compared with that in control group. Among the 20 largest genera in patients with ЕGE, significant abundance was observed for Lachnospira, Blautia, Dorea, Streptococcus, Eubacterium hallii_group paralleled with significant decline in Bifidobacterium and Eubacterium eligens_group. A positive correlation was obtained between estrogen levels and the number of representatives from the genera Eubacterium hallii_group and Streptococcus, IL-8 and Streptococcus, TNF-α and Streptococcus and Lachnospira, as well as a negative correlation between TNF-α and Bifidobacterium.Conclusion. A relation between dysbiotic intestinal alterations and developing endometriosis was found. The identified correlations between altered taxonomic composition of the intestinal microflora and parameters of hormonal and immune status in patients with ЕGE suggest that intestinal microbiota is involved in pathophysiology of endometriosis.

P-644 Effect of Letrozole co-stimulation with gonadotropin in women with normal ovarian reserve on oocyte maturation: a Randomized controlled trial

Abstract Study question To define whether letrozole co-stimulation with gonadotropin in normal-ovarian reserve women will improve the number of oocyte maturation. Summary answer Letrozole co-stimulation in normal ovarian reserve resulted in a reduced number of MII. What is known already Letrozole functions as an aromatase inhibitor. By inhibiting this enzyme, letrozole effectively lowers estrogen levels. Thus, letrozole prevents the estrogen-negative feedback to the HPO axis, which can increase the secretion of gonadotropins from the pituitary gland to promote folliculogenesis. The application of letrozole co-stimulation in women with a normal ovarian reserve is still debated. Study design, size, duration This study is an open-labeled, randomized controlled trial. Letrozole co-stimulation with gonadotropin in women with normal ovarian reserve. The RCT was conducted at the Fertility and Reproductive Biology Unit, Department of Obstetrics and Gynecology, Faculty of Medicine, Siriraj Hospital in Thailand, from March 2023 to October 2023. Participants/materials, setting, methods One hundred forty-four infertile women 18-40 years old with normal ovarian reserve (AMH 1.2-7 ng/ml, AFC ≥ 5) were random to gonadotropin co-stimulation with letrozole 5 mg once daily (n = 72) and gonadotropin alone (n = 72). The number of MII oocytes, follicle-to-oocyte index, GDF-9 and BMP-15 in follicular fluid were assessed. Main results and the role of chance Letrozole co-stimulation resulted in a significantly reduced number of MII [8 (4,12) vs 9 (5, 15.8), p = 0.042] and the rate of MII [ 71.7(60, 83.8) vs 80 (66.7, 90), p = 0.013]. However, the number of retrieved oocytes [12 (6,16.8) vs. 12 (6,16), p = 0.335] and FOI [85.7 (57.4,107.5) vs 84.2 [61.8, 112.5], p = 0.396] remained unchanged. The total dose of gonadotropin in the letrozole group was no different from the control group [1,350 (1,200, 1,650) vs 1,500 (1,350, 1,718.8) I.U., p = 0.137]. The duration of ovarian stimulation was shortened in the letrozole group compared to the control group [9 (8, 10) vs 9 (8.3, 10) days, p = 0.027]. The letrozole and control group had a similar level of GDF-9 (0.036 ± 0.003 A.U. vs. 0.028 ± 0.004 A.U., p = 0.119, letrozole and control group, respectively) and BMP-15 (0.034 ± 0.002 A.U. vs. 0.033 ± 0.004 A.U., p = 0.923, letrozole and control group respectively) in follicular fluid. Limitations, reasons for caution This study was open-label, with limited generalizability due to 91% of ovarian stimulation being strict to the research protocol (r-FSH and antagonist protocol) and no pregnancy outcome. Wider implications of the findings The study showed no benefit of letrozole co-stimulation in women with normal ovarian reserve. Trial registration number TCTR20221125002

O-279 The lower reproductive tract microbiota composition alters upon ovarian stimulation and is associated with, but not accurately predictive of reproductive outcomes following ART

Abstract Study question Are ovarian stimulation (OS) and ART success associated with the microbiota of the lower reproductive tract (LRT), i.e. vagina and cervix? Summary answer Estrogen rise during OS has a pivotal role in the LRT microbiota variation. The LRT microbiota pre-OS, although associated, may not accurately predict ART success. What is known already OS is a key component to enhance success rates in ART. OS triggers a significant rise in serum estrogen levels, which are known to influence the vaginal microbiota. The role of the female LRT microbiota in ART success is increasingly recognized, yet the impact of OS, particularly the estrogen rise, on these microbial communities and how the change may influence ART success remains largely unknown. Additionally, finding putative biomarkers within LRT microbiota could aid in predicting ART success. Understanding this relationship could refine and personalize fertility treatments, but robust evidence is needed to guide microbiota-based clinical decisions in ART. Study design, size, duration Vaginal and cervical swabs were collected longitudinally from patients undergoing ART and this at baseline (pre-OS) and at oocyte retrieval (post-OS). We gathered a final dataset comprising 275 vaginal samples at baseline and 240 samples at oocyte retrieval. Additionally, we defined a prospective cohort of n = 228x2 samples corresponding to patients for which both pre- and post-OS samples were available. The main results as detected within the vaginal samples were confirmed in the cervical samples. Participants/materials, setting, methods Bacterial DNA from −80 °C preserved swabs was extracted in a Tecan Nucleic Acid Extraction Platform, using the QIAamp DNA-Stool-Mini-Kit. Samples for paired-end Illumina MiSeq were constructed using a two-step PCR amplicon approach targeting the V4 region of the 16S rRNA. Then, LotuS, DADA2, the RDP classifier trainset 16 and variance stabilisation (poscounts, DESeq2) were used for read de-multiplexing, pre-processing, taxonomy assignment and transformation, respectively, as recommended for zero-inflated compositional data. Main results and the role of chance First, a confounder analysis revealed that both the menstrual phase and intercourse correlated with the LRT’s baseline microbiota composition (multivariate distance-based redundancy analysis [dbRDA], AdjP<0.05, N = 275). Next, in the paired cohort, we observed that the OS-associated rise in estrogen was correlated with confounder-independent compositional (confounder-adjusted dbRDA, AdjP=0.002, R2=1.14%, N = 228x2) and microbial-feature changes (Wilcoxon test, AdjP<0.05, Ndelta=228) in the LRT. Interestingly, post-OS we noted a decrease in Lactobacillus proportions compared to baseline (in participants sampled at either the follicular or luteal phase of the menstrual cycle). Additionally, post-OS microbiota composition was not associated with the live birth rate (LBR, following the first embryo transfer of the cycle) or the cumulative LBR (CLBR, following all embryo transfers of the cycle), nor with the total number of high-quality (HQ) embryos available (dbRDA, AdjP>0.05, N = 240). Notably, pre-OS microbiota communities and LRT Anaerococcus and Peptoniphilus proportions were associated with (C)LBR (confounder-adjusted dbRDA, AdjP<0.05; Logistic regression, odds-ratio<1, AdjP<0.05, N = 275). Machine learning models determined that vaginal Peptoniphilus and Anaerococcusproportions alone did not have good accuracy/ROC for predicting (C)LBR (test-set;<70%). Notably, the number of HQ embryos showed good accuracy/ROC for CLBR (test-set; 80%/84.5%). Including vaginal Anaerococcus proportions only slightly improved the model’s accuracy/ROC (test-set; 82.5%/85.6%). Limitations, reasons for caution The data’s proportional nature restricted assessing whether the estrogen rise decreased Lactobacillus proportions or increased other bacteria. Luteal phase sampling was avoided given the intravaginal progesterone support. Larger datasets may further improve predictive models. Wider implications of the findings Our study highlights the confounding impact of baseline variables and the influence of the OS-associated estrogen rise on LRT microbiota. It questions the clinical relevance of the LRT microbiota’s impact on reproductive outcomes following ART, warning for caution in microbiota-based decisions in ART. Trial registration number NCT03105453

Changes in Rehmanniae Radix processing and their impact on ovarian hypofunction: potential mechanisms of action.

This study evaluates the research developments concerning Rehmanniae Radix in ovarian hypofunction diseases. It explores the processing methods of Rehmanniae Radix, the variations in its compounds before and after processing, the mechanism of Rehmanniae Radix and its active compounds in improving ovarian function, and the advancements in clinical applications of traditional Chinese medicine (TCM) compound that include Rehmanniae Radix. Comprehensive literature search was conducted using databases such as China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database, National Science and Technology Library, the Pharmacopoeia of the People's Republic of China, Pubmed, and the Web of Science Database. The search utilized the following Medical Subject Headings (MeSH) and keywords: "Rehmanniae Radix," "Drying Rehmannia Root," "Rehmannia glutinosa," "Rehmanniae Radix Praeparata," "Traditional Chinese Medicine Processing," "Pharmacological Effects," "Ovarian Aging," "Diminished ovarian reserve," "Premature ovarian insufficiency," "Premature Ovarian Failure," "Ovarian hypofunction diseases". The ancient Chinese medical books document various processing techniques for Rehmanniae Radix. Contemporary research has identified changes in its compounds processing and the resultant diverse therapeutic effects. When processed into Rehmanniae Radix Praeparata, it is noted for its ability to invigorate the kidney. TCM compound containing Rehmanniae Radix is frequently used to treat ovarian hypofunction diseases, demonstrating significant clinical effectiveness. The key changes in its compounds processing include cyclic dilute ether terpene glycosides, phenylethanol glycosides, sugars, and 5-hydroxymethylfurfural. Its pharmacological action is primarily linked to the improvement of granulosa cell proliferation, antioxidative and anti-aging properties, and modulation of the immune and inflammatory microenvironment. Furthermore, Rehmanniae Radix also offers therapeutic benefits for cardiovascular and cerebrovascular diseases, osteoporosis and cognitive dysfunction caused by low estrogen levels. Thereby Rehmanniae Radix mitigates both the short-term and long-term health risks associated with ovarian hypofunction diseases. Processed Rehmanniae Radix has shown potential to improve ovarian function, and its compound prescriptions have a definite effect on ovarian dysfunction diseases. Therefore Rehmanniae Radix was garnering interest for both basic and clinical research, with promising application prospects as a future therapeutic agent for ovarian hypofunction diseases. However, further studies on its toxicology and the design of standardized clinical trials are necessary to fully establish its efficacy and safety.

O-213 Analysis of risk factors for monozygotic twins following in vitro fertilization-embryo transfer

Abstract Study question To investigate the risk factors associated with the incidence of monozygotic twinning（MZT）following in vitro fertilization and embryo transfer (IVF-ET). Summary answer MZT following IVF-ET may be associated with a reduced endometrial thickness on the day of embryo transfer and a decrease in estradiol levels post-transfer. What is known already MZT pregnancy arises from the division of a single fertilized ovum, resulting in offspring with identical genetic material. These twins exhibit markedly higher rates of complications compared to dizygotic counterparts, including increased incidences of miscarriage, congenital anomalies, intrauterine fetal demise, growth restriction, and preterm delivery. The underlying biological processes leading to MZT remain elusive. Presently, scholarly discourse centers around two predominant models and one hypothesis: the zygote division model, the embryonic membrane fusion model, and the over-mature gametes hypothesis, respectively. IVF-ET has been implicated in the heightened occurrence of MZT, suggesting a potential iatrogenic element to this phenomenon. Study design, size, duration A retrospective cohort study has been conducted. We analyzed 582 IVF-ET cycles at a large teaching hospital from January 2007 to December 2022, including 291 clinically confirmed MZTs, and 291 matched non-MZTs. Participants/materials, setting, methods A total of 582 participants, 291 in both MZT group and non-MZT group. There were 146 fresh embryo transfer cycles and 145 frozen-thawed cycles in the MZT group. Each cycle type was matched on a 1:1 basis by age and embryo transfer date with a control group that did not result in MZT following IVF-ET. We examined the disparities between the MZT group and the non-MZT group, and performed the Logistics regression analysis. Main results and the role of chance Significant differences were observed in the MZT group compared to the non-MZT group in the fresh embryo transfer cycle, particularly regarding estradiol (E2) and progesterone (P) levels on day 7 post-transfer, E2/P ratios on day 14 post-transfer, and endometrial thickness on the day of transfer (p<0.05). Logistic regression analysis indicated a correlation between reduced endometrial thickness on the transfer day and the occurrence of MZT. During the frozen-thawed cycle, significant differences were noted in E2/P ratios, E2 levels on day 7 post-transfer, and endometrial thickness on embryo transfer day (p<0.05). Logistic regression analysis suggested that lower E2 levels on day 7 post-transfer and reduced endometrial thickness on embryo transfer day were predictive of MZT. Additional potential influencing factors included the duration of the participants’ menstrual cycle, the protocol of ovulation induction, and the duration of embryo culture in vitro. Limitations, reasons for caution Primarily, the retrospective design may introduce bias, notably due to the potential absence of comprehensive data. Secondly, the study is constrained by the sample size, with 146 fresh cycles and 145 frozen cycles, which may not provide a fully representative statistical power. Wider implications of the findings The incidence of MZT may implicate signaling pathways at the maternal-fetal interface, with early embryonic split potentially serving as a compensatory mechanism in response to suboptimal estrogen levels or diminished endometrial thickness, thus enhancing embryonic viability. Trial registration number not applicable

Hormone replacement therapy did not alleviate temporomandibular joint inflammatory pain in ovariectomized rats.

This study had the aim of examining the relationships between variations in estrogen levels resulting from ovariectomy, and estrogen hormone replacement therapy (HRT) in rats subjected to an orofacial inflammatory pain model. Eighty adult female Wistar rats were initially divided into 2 groups: Sham or ovariectomy (OVX-D1). Seven days later (D7), the rats were subjected to an unilateral infiltration of Freund's Complete Adjuvant (CFA) or saline solution into the right temporomandibular joint (TMJ). Then, rats received 17β-estradiol (28µg/kg/day) or placebo for 21days (D10-D31). Nociception was evaluated by the von Frey (VF) and the Hot Plate (HP) tests, and depressive-like behavior by the Forced Swimming (FS) test. On D32 all rats were euthanized and serum, hippocampus and brainstem were collected. The CFA groups presented a mechanical hyperalgesia until day 21 (p ≤ 0.05). No differences were observed among groups in the HP (p = 0.735), and in the immobility and swimming time of the FS (p = 0.800; p = 0.998, respectively). In the brainstem, there was a significant difference in the TNF-ɑ levels (p = 0.043), and a marginal significant difference in BDNF levels (p = 0.054), without differences among groups in the hippocampal BDNF and TNF-ɑ levels (p = 0.232; p = 0.081, respectively). In conclusion, the hormone replacement therapy did not alleviate orofacial pain in ovariectomized rats. However, there is a decrease in brainstem TNF-ɑ levels in the animals submitted to both models, which was partially reverted by HRT.

The Comparative Study of Serum Estrogen and Lipid Profile in Pre- and Post-menopausal Women as Atherosclerosis Risk Factors in Pakistan.

Menopausesignifies the eternal termination of menstruation in women as a consequence of ovarian action loss, typically occurring around the age of 51 years. Cardiovascular disease is the leading cause of death among post-menopausal women, which may be due to lower levels of estrogen and lipid profile. The present study was undertaken to evaluate serum estrogen and lipid profile status to assess the risk of atherosclerosis in both pre- and post-menopausal women. The objective of this study is to explorethe relationship between estrogen and lipid levels of women in pre- and post-menopausal stages. A comparative cross-sectional study was conducted at Railway General Hospital Rawalpindi. A total of 100 participants were included of which 50 were pre-menopausal and 50 were post-menopausal women. Laboratory examination and questionnaires from the study population were used for data collection. Through the enzymatic method, serum cholesterol, triglycerides, low-density lipoprotein, and high-density lipoprotein (HDL) were assessed. Serum very low-density lipoprotein (VLDL) levels were calculated via Friedwald's components VLDL=TG/5.0. An enzyme-linked immunosorbent assay kit was used for estrogen measurement. For statistical analysis, Student's t-test and the Pearson correlation test were used. Women after menopause have significantly high serum cholesterol, low-density lipoproteins, VLDLs, and triglycerides while HDL-c levels were significantly low (P<0.001). Levels of estrogen were low in post-menopausal females (P<0.001) as compared to menstruating women. Estrogen with HDL concentrations showed a positive correlation with an r value of 0.08556 while LDL levels showed a negative correlation with ar value of -0.26219. This comparative study explores the relationship between estrogen and lipid levels in pre- and post-menopausal women. Low estrogen with changed lipid variables was observed. Decreased cardiovascular protective HDL-c marks that menopause is a phase that acts as an independent risk factor for atherosclerosis.

The Therapy Based on Traditional Chinese Medicine for Otolithiasis

Background: Otolithiasis (also named benign paroxysmal positional vertigo, BPPV) is one of the most common illness for hospital visits. Residual dizziness and recurrence after reduction are problems for the treating BPPV with Western medicine. Traditional Chinese medicine (TCM) has shown obvious superiority in the treating BPPV. Here, we explore the curative effect of the theory of dizziness of TCM in treating BPPV. Methods: 183 BPPV patients were randomly divided into 3 groups: Reset group (RG) 61 cases, Traditional Chinese Medicine group (TG) 61 cases, and Reset plus Traditional Chinese medicine group (R+TG) 61 cases. RG received machine reset. TG was given the therapy based on the theory of dizziness of TCM. R+TG received machine reset plus the therapy of TCM. All patients, before and on 1th day, 7th day, and 14th day of treatment, were evaluated with the dizziness handicap inventory (DHI) for the physical (P), functional (F) and emotional (E) state. The efficacy of the different therapy in the treatment of mild, moderate, and severe BPPV has been observed. Follow up for 6 months, we record and compare the ratio of patients with residual dizziness and the recurrence of BPPV for the three groups. Serum levels of vitamin D, estrogen, and calcium and phosphorus ions were detected and compared between the groups of BPPV patients, dizziness non-BPPV patients, and non-dizziness patients. Results: The DHI score of the three groups, compared with which before treatment, RG (p &lt; 0.05), TG (p &lt; 0.01), and R+TG (p &lt; 0.01), decreased significantly on the 7th day of treatment; R+TG decreased more significantly than RG (p &lt; 0.05). The DHI scores of the three groups, on the 14th day of treatment, were significantly lower than those before treatment (p &lt; 0.01). On the 7th day of treatment, compared with the same period of RG, the score of item E of TG and R+TG, decreased significantly (p &lt; 0.05). Patients of mild BPPV responded well to the three kinds of the therapy; For the patients of moderate BPPV, on the 7th day of treatment, assessed by DHI, the effect showed significant improve for RG (p &lt; 0.05), TG (p &lt; 0.01) and R+TG (p &lt; 0.01). For the patients of severe BPPV, on the 7th day of treatment, the effect of RG, assessed with DHI, showed no improvement (p &gt; 0.05), while TG (p &lt; 0.05) and RG+TG (p &lt; 0.01) showed effective. The incidence of residual dizziness between the three groups show significantly different on the 2nd week (x2 = 7.635, p = 0.022) and the 1th month (x2 = 7.502, p = 0.023) of treatment; The incidence of RG was higher than those of TG and R+TG. The recurrence rates of BPPV of the three groups were significantly different on the end of 2nd month (x2 = 8.528, p = 0.014), 4th month (x2 = 13.287, p = 0.001), and 6th month (x2 = 12.587, p = 0.002) of treatment. The recurrence rates of TG and R+TG were lower than that of RG. There was no significant difference in recurrence rate and residual dizziness occurrence rate between TG and R+TG. There were differences in serum vitamin D and estrogen levels between the group of BPPV patients, dizziness non-BPPV patients, and non-dizziness patients; There was no difference in calcium and phosphorus ion levels between the three groups. Conclusions: Compared with therapy of RG, TG and R+TG showed more effective in treating BPPV; They can significantly improve patient's bad mood, reduce the proportion of residual dizziness, and reduce the recurrence rate. TCM and TCM plus reset have better efficacy in treating moderate and severe BPPV. The specificity of abnormal changes in serum vitamin D and estrogen for BPPV patients needs be further study.

Menopause and Estrogen Associations With Gut Barrier, Microbial Translocation, and Immune Activation Biomarkers in Women With and Without HIV.

Estrogens may protect the gut barrier and reduce microbial translocation and immune activation, which are prevalent in HIV infection. We investigated relationships of the menopausal transition and estrogens with gut barrier, microbial translocation, and immune activation biomarkers in women with and without HIV. Longitudinal and cross-sectional studies nested in the Women's Interagency HIV Study. Intestinal fatty acid binding protein, lipopolysaccharide binding protein, and soluble CD14 (sCD14) levels were measured in serum from 77 women (43 with HIV) before, during, and after the menopausal transition (∼6 measures per woman over ∼13 years). A separate cross-sectional analysis was conducted among 72 postmenopausal women with HIV with these biomarkers and serum estrogens. Women in the longitudinal analysis were a median age of 43 years at baseline. In piecewise, linear, mixed-effects models with cutpoints 2 years before and after the final menstrual period to delineate the menopausal transition, sCD14 levels increased over time during the menopausal transition (Beta [95% CI]: 38 [12 to 64] ng/mL/yr, P = 0.004), followed by a decrease posttransition (-46 [-75 to -18], P = 0.001), with the piecewise model providing a better fit than a linear model (P = 0.0006). In stratified analyses, these results were only apparent in women with HIV. In cross-sectional analyses, among women with HIV, free estradiol inversely correlated with sCD14 levels (r = -0.26, P = 0.03). Lipopolysaccharide binding protein and intestinal fatty acid binding protein levels did not appear related to the menopausal transition and estrogen levels. Women with HIV may experience heightened innate immune activation during menopause, possibly related to the depletion of estrogens.

Exploring the Intersection of Depression, Anxiety, and Sexual Health in Perimenopausal Women.

The perimenopausal period is marked by hormonal fluctuations that trigger a complex interplay between estrogen levels and neurotransmitters' function, contributing to increased susceptibility to depression and anxiety in women. Concurrently, hormonal changes, coupled with alterations in vaginal tissue, lead to sexual dysfunction during this transitional phase. This study aimed at evaluating the association between menopausal symptoms and sexual dysfunction among perimenopausal women and identifying the mediating effects of depression and anxiety on this association. Data for the present cross-sectional study were collected from participants via Arabic versions of three questionnaires; the modified Menopausal Rating Scale (MRS), the Female Sexual Functioning Index (FSFI) and the Hospital Anxiety and Depression scale (HADS). Our study was conducted on 149 females with age ranged from 45 to 55 years. On studying the relation between modified MRS and HADS, the menopausal symptoms were significantly high among female with high anxiety scores. Regarding the relationship between MRS and FSFI, women with anxiety and physical and mental exhaustion had significantly lower FSFI scores than women without such symptoms (19.2 [2-31.4] vs 21.7 [3.8-30.9], p = 0.04, respectively). Furthermore, there were statistically significant negative correlations between depression scores and sexual desire (r = -0.32, p < 0.001), arousal (r = -0.25, p = 0.003), and total FSFI scores (r = -0.27, p = 0.04). Perimenopausal women experience a confluence of challenges related to depression, anxiety, and sexual dysfunction. Understanding the interconnectedness of hormonal and psychosocial factors is essential for tailored interventions aimed at improving mental health and sexual well-being during this transitional phase.