Estradiol Research Articles

¿Qué sabemos de los esteroides sexuales y las gonadotropinas en la reproducción de teleósteos neotropicales?

Esta revisión provee un resumen general de las investigaciones realizadas en los últimos 10 años acerca de los esteroides sexuales (17β-Estradiol, Testosterona, 11 Ketotestosterona y 17α, 20β-DHP), las gonadotropinas (Fsh y Lh) y sus principales reguladores (GnRH1-3 y GnIH1-3); y su respectiva función en la modulación de la reproducción. Específicamente sobre el eje Hipotálamo – Pituitaria – Gónada (HPG), en teleósteos modelo y neotropicales (Astyanax altiparanae, Steindachneridion parahybae, Salminus hillarii, Centropomus undecimalis). Mostramos que el papel regulador de éstos continúa siendo tema de discusión, pues varía en función de múltiples características, entre ellas, las estrategiareproductivas de la especie en cuestión y/o las condiciones en las que éstas son mantenidas durante el estudio. Históricamente, se ha descrito que la Fsh actúa en las fases tempranas de la reproducción, estimulando la biosíntesis de estradiol y testosterona necesarios para la gametogénesis; mientras la Lh está implicada en las fases finales del ciclo reproductivo (maduración final de los gametos y desove). Sin embargo, en especies con desarrollo asincrónico y desove múltiple, la Lh se produce durante todo el ciclo en paralelo con la Fsh, sugiriendo la necesidad de ambas hormonas en la gametogénesis y maduración final. Adicionalmente, investigaciones recientes en especies con comportamiento migratorio reproductivo, han evidenciado una disminución significativa en la expresión del ARNm de la subunidad β de Lh (lhb) cuando los individuos son mantenidos en cautiverio, lo que podría ocasionar alguna disfunción reproductiva bajo los actuales sistemas de cultivo, considerado hoy el gran problema de la acuicultura contemporánea. Desafortunadamente, los estudios de estos tópicos en especies suramericanas son aún incipientes, por lo que se hace necesario centrar las investigaciones hacia el esclarecimiento del control neuroendocrino de la reproducción en especies nativas, más aún cuando estas especies son mantenidas en cautiverio.

The Impact of Prenatal Exposure to Bisphenol A on Male Reproductive Function.

Bisphenol A (BPA) is a recognized xenoestrogen, in that it possesses oestrogenic and anti-androgenic properties. These endocrine-disrupting effects of BPA at the estrogen receptor (ER) occur despite the very low affinity of BPA for the ERβ, which is 10,000 times lower than that of 17-β estradiol, and despite the European regulatory authorities stating that BPA is safe, at usual exposure concentrations, the use of BPA in baby drink bottles was banned in 2011. There exists conflicting evidence from human epidemiological studies as to its influence on adult male reproductive function, although animal data is more convincing. This mini-review will report on the limited epidemiological data from human studies relating early life exposure to BPA on adult male reproductive function. A long term follow-up study from Western Australia using a birth cohort, the Raine Study, demonstrated no adverse associations of antenatal exposure to BPA, and potentially a positive association with antenatal BPA exposure with sperm concentration and motility at 20 years of age, although recent scientific reports suggest traditional measures of BPA exposure may underestimate exposure levels, which makes data interpretation potentially flawed.

Biocompatible amino acid-based ionic liquids for extracting hormones and antibiotics from swine effluents

The generalized use of antibiotics and hormones in swine feed has triggered a growing concern on their presence in wastewater effluents and manure. Therefore, the implementation of strategies for their removal is a matter of practical significance and the synthesis of biocompatible ionic liquids to be used as segregation agents in aqueous waste effluents could be a suitable means for removing these emerging contaminants. In this work, the salting out capacity of cholinium alaninate, (N1112OHAla) in aqueous streams containing surfactants (Triton X-100) was demonstrated. Afterwards, its ability to separate a hormone (17-β estradiol) and antibiotics belonging to tetracycline and fluoroquinolone families was investigated in an aqueous synthetic swine effluent at environmental conditions. It was checked that the cautious selection of the feed compositions allowed high levels of hormone extraction (E) and partition coefficients (K): E > 90% and K > 13.86, in surfactant rich-phase. Antibiotics migrated to any of the two phases depending on system compositions. However, the decisive addition of a cationic surfactant (2% CTAB) led to increased extraction efficiencies of chlortetracycline (K = 15.77 and E = 92.5%), proving the versatility of the process.

Pharmacological Modulation of Steroid Activity in Hormone-Dependent Breast and Prostate Cancers: Effect of Some Plant Extract Derivatives.

The great majority of breast and prostate tumors are hormone-dependent cancers; hence, estrogens and androgens can, respectively, drive their developments, making it possible to use pharmacological therapies in their hormone-dependent phases by targeting the levels of steroid or modulating their physiological activity through their respective nuclear receptors when the tumors relapse. Unfortunately, at some stage, both breast and prostate cancers become resistant to pharmacological treatments that aim to block their receptors, estrogen (ER) or androgen (AR) receptors, respectively. So far, antiestrogens and antiandrogens used in clinics have been designed based on their structural analogies with natural hormones, 17-β estradiol and dihydrotestosterone. Plants are a potential source of drug discovery and the development of new pharmacological compounds. The aim of this review article is to highlight the recent advances in the pharmacological modulation of androgen or estrogen levels, and their activity through their cognate nuclear receptors in prostate or breast cancer and the effects of some plants extracts.

In vitro oocyte maturation and blood‐circulating steroid hormones in Sterlet ( Acipenser ruthenus ) and Siberian sturgeon ( Acipenser baeri )

The oocyte of Sterlet (Acipenser ruthenus) and Siberian sturgeon (Acipenser baeri) was evaluated after incubation with 17α, 20β-Dihydroxyprogesterone (DHP) in artificial media. Concentration of testosterone (T), progesterone (P4) and 17-β estradiol (E2) in blood and their relation to fertilization rate were measured during experimental period. The oocytes at the same developmental stages were incubated in SIS (based on sturgeon blood ionic composition), RM2 (Ringer solution modified for sturgeons) and L-15 (Leibovitz medium) artificial mediums in the presence of 1 μg/ml of 17α, 20β-Dihydroxyprogesterone for 12, 18 and 24 hr. The result of this study demonstrated that fertilization rate decreased with the increasing duration of incubation in all mediums. In the Siberian and Sterlet sturgeon, the highest fertilization rate was observed in the oocytes incubated in RM2 (0.81 ± 0.4) and SIS (0.44 ± 0.8) mediums for 12 hr respectively. In the Siberian sturgeon, fertilization rate decreased significantly (p < .05) with incubation time in all three mediums. In Starlet, fertilization rate decreased significantly (p < .05) with time and reached a minimum after 24 hr (0.16 ± 0.1) in L-15 medium. No significant (p > .05) differences were observed between blood plasma hormones and fertilization rate. The results of this study indicated that 17α, 20β-Dihydroxyprogesterone is essential for induction of in vitro oocyte maturation in these species.

Hormonal and biochemical studies on female dromedary camels affected with multiple ovarian cysts

Abstract The purpose of the present study was to investigate the hormonal and biochemical profile of dromedary she-camels with multiple ovarian cysts (MOC). Seventeen she-camels suffering from MOC and 10 control animals were enrolled in this study. Blood samples were collected from all animals. Progesterone (P4), estradiol 17-β (E2), Testosterone, tri-iodothyronine (T3), Thyroxine (T4), glucose, total protein, albumin, total cholesterol, triglycerides, HDL, nitric oxide (NO), malondialdehyde (MDA) and glutathione peroxidase (GPX) levels were estimated in blood serum. Results revealed the presence of three or more large follicular ovarian cysts either on one (n = 6) or both ovaries (n = 11) of MOC cases with absence of any luteal structure. She-camels with multiple MOC had significantly (P ≤ 0.001) lower levels of P4 (0.24 ± 0.02 vs. 0.40 ± 0.03 ng/mL, respectively) and significantly higher levels of E2 and T3 (19.62 ± 0.62 vs. 13.74 ± 0.67 pg/mL, P ≤ 0.001 and 1.85 ± 0.09 vs. 1.44 ± 0.10 ng/mL, P ≤ 0.012, respectively) as compared with control. There were no significant differences between the two groups regarding testosterone and T4 levels (0.031 ± 0.002 vs. 0.026 ± 0.001 and 10.93 ± 0.24 vs. 10.17 ± 0.28 ng/mL, respectively). She-camels having MOC had significantly higher levels of glucose, cholesterol, triglycerides, HDL, total lipids and MDA and lower levels of GPX and NO as compared with control group. There were no significant differences between the two groups in levels of total proteins, albumin and globulin. In conclusion, the hormonal imbalances, metabolic disorders and oxidative stress are involved in the formation and/or persistence of multiple ovarian cysts in dromedary camels.

Estrogen increases expression of vascular alpha 2C adrenoceptor through the cAMP/Epac/JNK/AP-1 pathway and potentiates cold-induced vasoconstriction

Cutaneous cold-induced vasoconstriction is a normal physiological reaction mediated by alpha 2C-adrenergic receptors (α2C-ARs) expressed in vascular smooth muscle cells (VSMCs). When this reaction is exaggerated, Raynaud's phenomenon (RP) ensues. RP is more prevalent in females compared to age-matched men. We previously established that 17-β estradiol (estrogen) upregulates α2C-ARs in human VSMCs via a cAMP/Epac/Rap pathway. We also showed that cAMP acts through JNK to increase α2C-AR expression. However, whether estrogen employs JNK to regulate α2C-AR is not investigated. Knowing that the α2C-AR promoter harbors an activator protein-1 (AP-1) binding site that can be potentially activated by JNK, we hypothesized that estrogen regulates α2C-AR expression through an Epac/JNK/AP-1 pathway. Our results show that estrogen (10−10 M) activated JNK in human VSMCs extracted from cutaneous arterioles. Pretreatment with ESI09 (10 μM; an Epac inhibitor), abolished estrogen-induced JNK activation. In addition, pre-treatment with SP600125 (3 μM; a JNK specific inhibitor) abolished estrogen-induced expression of α2C-AR. Importantly, estrogen-induced activation of α2C-AR promoter was attenuated with SP600125. Moreover, transient transfection of VSMCs with an Epac dominant negative mutant (Epac-DN) abolished estrogen-induced activation of α2C-AR promoter. However, co-transfection of constitutively active JNK mutant overrode the inhibitory effect of Epac-DN on α2C-AR promoter. Moreover, estrogen caused a concentration-dependent increase in the activity of AP-1-driven reporter construct. Mutation of AP-1 site in the α2C-AR promoter abolished its activation by estrogen. This in vitro estrogen-increased α2C-AR expression was mirrored by an increase in the ex vivo functional responsiveness of arterioles. Indeed, estrogen potentiated α2C-AR-mediated cold-induced vasoconstriction, which was abolished by SP600125. Collectively, these results indicate that estrogen upregulates α2C-AR expression via an EPAC-mediated JNK/AP-1- dependent mechanism. These results provide an insight into the mechanism by which exaggerated cold-induced vasoconstriction occurs in estrogen-replete females and identify Epac and JNK as potential targets for the treatment of RP.

Luteinizing hormone and ovarian steroids affect in vitro prostaglandin production in the equine myometrium and endometrium

Prostaglandins (PGs) play crucial roles in the regulation of the oestrus cycle and establishment of pregnancy in animals. Luteinizing hormone (LH) and ovarian steroids are involved in regulating endometrial PG production in many species. Their effects on PG production and associated pathways in the mare myometrium and endometrium are the subjects of our interest. This study aimed to evaluate the specific effects of LH and ovarian steroids on equine myometrial and endometrial tissues on (i) PGE2 and PGF2α secretion and (ii) transcription of genes encoding specific enzymes responsible for PG synthesis, such as prostaglandin–endoperoxide synthase (PTGS2), PGE2 synthases (PGES), PGF2α synthases (PGFS), and PGI2 synthases (PGIS), using equine myometrial and endometrial explants. Equine myometrial and endometrial tissues were collected at the mid-luteal (n = 6) and follicular (n = 6) phases of the oestrus cycle and were exposed to: (1) vehicle (control), (2) arachidonic acid (AA, 50 ng/mL, positive control), (3) LH (10 ng/mL), (4) progesterone (P4, 10−7M) and (5) 17-β oestradiol (E2, 10−9M) for 24 h. After exposure, PGF2α and PGE2 concentrations were determined using direct enzyme immunoassays. Alterations in PG synthase mRNA expression were determined using RT-qPCR. After 24 h, LH and P4 increased PGE2 and PGF2α secretion by myometrial tissues at the mid-luteal phase (P < 0.05), whereas PG secretion was augmented by LH and E2 during the follicular phase (P < 0.01). In contrast, LH and E2 increased PGE2 and PGF2α secretion by endometrial tissues during the mid-luteal phase (P < 0.05), while E2 enhanced PGE2 secretion during the follicular phase of the oestrus cycle (P < 0.01). These results indicate that LH and ovarian steroids modulate PG production in equine myometrial and endometrial tissues and affect PG synthase expression at the mRNA level. We conclude that the equine myometrium is an alternative source of PG production and participates in the regulation of uterus function during the oestrus cycle.

The effect of female sex steroid hormones on osteogenic differentiation of endometrial stem cells.

Bone regeneration is a significant and crucial health issue worldwide. Tissue bioengineering has shown itself to be the best substitute for common clinical treatment of bone loss. The suitable cell source is human endometrial stem cells (hEnSCs) which have several suitable characteristics for this approach. Since sex steroid hormones are involved in expansion and conservation of the skeleton, the effect of two sex steroid hormones known as estrogen (17-β estradiol) and progesterone on osteogenic differentiation of hEnSCs were examined. For this purpose, hEnSCs were treated with 17-β estradiol and progesterone separately (1 × 10-6 M) and simultaneously (1 × 10-7 M). Osteogenic differentiation tests including measurement of total mineral calcium content, Alizarin Red staining, the quantitative expression levels of some osteogenic markers by Real-time RT-PCR, and immunofluorescence staining were performed at 7 and 14days of differentiation. To exhibit the morphology of the cells in osteogenic and culture medium, the hEnSCs were stained with Acridine Orange (AO) solution. In this research, MTT assay and AO staining revealed progesterone and 17-β estradiol increase the proliferation of hEnSCs in a dose-dependent manner. Furthermore, the results of calcium content analysis, Real-time RT-PCR assay, and all tests of differentiation staining have shown that 17-β estradiol and progesterone cannot induce hEnSCs' osteogenic differentiation. In conclusion, it is indicated that 17-β estradiol and progesterone do not have positive effects on hEnSCs' osteogenic differentiation in vitro.

Effect of supplementing additives in leptin-enriched maturation medium during in vitro maturation and vitrification of goat oocytes

The present study aimed to evaluate the effect of supplementing epidermal growth factor EGF , insulin-like growth factor-I IGF-I , fetal bovine serum FBS , and bovine serum albumin BSA in different combinations in a maturation medium MM comprising TCM-199 + 5 µg/mL 17-β estradiol + 5 µg/mL ovine-luteinizing hormone oLH + 5 µg/mL pure follicle-stimulating hormone pFSH + 100 µM/mL cysteamine containing the best concentration of leptin among 10/20/25/30 ng/mL on in vitro maturation IVM and subsequent vitrification of matured goat oocytes. The IVM of oocytes was carried out in a CO2 incubator for 27 h at 38.5 °C. The percentage of oocytes with cumulus expansion and polar body extrusion was found to be highest in 25 ng/mL leptin at 47.37% and 15.63%; rates of 66.67% and 28.57% were obtained when supplemented with 50 ng/mL IGF-I + 10 ng/mL EGF in medium containing 25 ng/mL leptin as compared to the other media. The in vitro matured oocytes were vitrified in 5 M ethylene glycol + 5 M propylene glycol with 0.5 M sucrose in TCM-199 + 20% FBS using liquid nitrogen. The highest percentage of morphologically intact in vitro matured oocytes following vitrification was obtained in medium containing 25 ng/mL leptin 89.29% . It was concluded that addition of IGF-I + EGF with 25 ng/mL leptin in MM yielded a higher rate of maturation of oocytes, and supplementation of 25 ng/mL leptin in MM resulted in a higher rate of morphologically intact vitrified oocytes, indicating the efficiency of leptin for cryotolerance.

Comparative analysis of the chemical constituents from the tuberous root and stem of Pueraria candollei var. mirifica and evaluation of their estrogenic activity

Background: Pueraria candollei var.mirifica (PC) is a tuberous plant enriched with bioactive phytoestrogens. PC is currently high demanded in the global markets as dietary supplement products, which resulted in a shortage of tuberous root from the natural resource and the field cultivation. Materials and Methods: We compare phytoestrogens contents from the tuberous root and stem of PC and their estrogenic activities on MCF-7 cells proliferation. Results: The root and stem of PC accumulated different contents of phytoestrogens. The highest amount of total isoflavonoids and total chromenes was found in the root bark of PC. However, the stem bark (10 μg/mL crude extract) which contains a high level of chromenes and low amount of isoflavonoids exhibited higher growth stimulation of MCF-7 cells (127.25% ± 4.43% relative proliferative effect [RPE] as compared to estradiol [10−10 M]) than root bark at the same concentration with 116.32% ± 3.59% RPE. After the solubilization of the ethanol extract with 10% (v/v) aqueous ethanol, the soluble fraction of crude extract of stem stimulated proliferation of MCF-7 cells without cell growth suppression effect at high concentration (100 μg/mL) as observed in the insoluble fraction of the extract. Conclusion: We suggested that the soluble part of crude extract might be less toxic than the ethanolic crude extract and its non-polar components. Our study opens the possibility of using the stem of PC as a new alternative source of bioactive phytoestrogens for dietary supplement products.

Estradiol’s Permissive Role in Social Stress Susceptibility in Female Rats

Social stress is a common risk factor for both depression and cardiovascular disease (CVD), and individuals suffering from depression are at increased risk of CVD‐related mortality. Additionally, women are more likely than men to suffer from depression and anxiety along with comorbid CVD, however this increased risk is confined to the range between the beginning of puberty and end of menopause. Therefore, while specific systems regulating heightened susceptibility in females are unclear, there is evidence to suggest it may be regulated in part by ovarian hormones. Using a social stress model in female rats whereby a female is forced to witness a social defeat encounter between two males (15 mins, 5 days), we have previously shown that intact females are highly susceptible to this form of stress as evidenced by anxiety‐and depressive‐like behaviors, brain cytokines, and diminished vagal tone. Importantly, ovariectomized (OVX) females are resistant. Here we conducted two separate sets of studies to determine the role that 17‐β estradiol (17‐βE, 10μg/rat, sc) plays in susceptibility to social stress in females. In study #1, following ovariectomy and surgical implantation of cardiovascular transmitters (HD‐S11, DSI), rats were treated with either 17‐βE or vehicle and subjected to control or witness stress during cardiovascular recording. In study #2 all females were implanted with an i.v. catheter and either left intact (sham, and treated with vehicle) or OVX (and treated with vehicle or 17‐βE), followed one hour later by witness stress or control handling. Blood samples were collected (0, 15 and 60 mins) and behavior was video recorded. OVX females exhibited a blunted tachycardic response to stress that was reversed by 17‐βE administration. Moreover, epinephrine release in response to witness stress was highly reactive in intact females above that of OVX females, despite similar corticosterone levels. Anxiety‐like responses were also shown to be regulated by 17‐βE; witness stress‐evoked burying that emerged in intact females was blunted by OVX and reinstated by 17‐βE. Taken together these studies identify that proestrous levels of estradiol are critical for acute behavioral and cardiovascular responses to social stress in females. Future studies are aimed at determining the neural systems that are regulated by estradiol and precipitate stress susceptibility.Support or Funding InformationMH113892‐01A1, NARSAD Young Investigator Award 26809

Association between Urinary Metabolite Levels of Organophosphorus Flame Retardants and Serum Sex Hormone Levels Measured in a Reference Sample of the US General Population.

Use of organophosphorus flame retardants (OPFRs) in consumer materials have led to widespread human exposure.Research is needed to examine the health effects attributable to the general population's exposure to OPFRs. Using the data from the National Health and Nutrition Examination Surveys (NHANES) (2013-2014), multiple regression analyses were performed to compare the adjusted geometric means (aGMs) of serum sex hormone by quartiles of urinary metabolites of OPFRs, including diphenyl phosphate (DPhP), bis(1,3-dichloro-2-propyl) phosphate (BDCPP), bis(2-chloroethyl) phosphate (BCEP) and dibutyl phosphate(DBuP), in children (6 - 9 years old), adolescents (10 - 19 years old) and adults(≥ 20 years old), while accounting for potential confounding factors. The aGMs of sex hormone-binding globulin increased by 36% (95% CI: 6.1 - 56.7%) in female children (p = 0.03), 44% (95%CI: 16 - 63%) in female adolescents (p = 0.010), and 22% (95%CI: 3.51 - 37%) in female adults (p = 0.025), from the 1st to 4th quartiles of the levels of DPhP, BDCPP, DBUP, respectively. The aGMs of estradiol (EST) decreased by 64% and 77% from the 1st to 4th quartiles of the DBUP levels in female children (p = 0.015) and female adolescent (p = 0.020), respectively. The aGMs of EST increased by 31% (95%CI: 3.8 - 51%) from the 1st to 4th quartiles of the DBUP levels in female adults (p = 0.031). These findings suggest that exposure to certain OPFRs is associated with the altered sex hormone levels in this sample of US population. More studies are needed to examine the mechanisms responsible for these observations.

Type 5 17-hydroxysteroid dehydrogenase/prostaglandin F synthase (AKR1C3) inhibition and potential anti-proliferative activity of cholest-4-ene-3,6-dione in MCF-7 breast cancer cells

Cholest-4-ene-3,6-dione (KS) is a cholesterol oxidation product which exhibits anti-proliferative activity. However, its precise mechanism of action remains unknown. In this study, the effects of KS on AKR1C3 inhibition and anti-proliferative activities were investigated in the hormone-dependent MCF-7 breast cancer cells. We identified that KS arrested the enzymatic conversion of estrone to 17-β estradiol, by inhibiting AKR1C3 in intact MCF-7 cells. The anti-proliferative effects of KS were evaluated by MTT assay, acridine orange and ethidium bromide dual staining, cell cycle analysis and Western blotting. KS arrested the cell cycle progression in the G1 phase with a concomitant increase of the Sub-G0 population to increase in concentration and time. It also enhanced the p53 and NFkB expression and induced caspase-12, 9 and 3 processing and down-regulated the Bcl-2 expression. Molecular docking studies performed to understand the inhibition mechanism of KS on AKR1C3 revealed that KS occupied the binding region of AKR1C3 with almost similar orientation as indomethacin (IM), thereby acting as an antagonistic agent for AKR1C3. Based on the results it is identified that KS induces inhibition of AKR1C3 and cell death in MCF-7 cells. These results indicate that KS can be used as a molecular scaffold for further development of novel small-molecules with better specificity towards AKR1C3.

Clinical effect of ethinylestradiol cycloproterone combined with metformin in the treatment of PCOS patients with insulin resistance

Objective To investigate the clinical effect of ethinylestradiol cycloproterone combined with metformin in the treatment of polycystic ovary syndrome (PCOS) with insulin resistance. Methods Ninety PCOS patients with insulin resistance admitted to the First People's Hospital of Taizhou from May 2017 to May 2019 were selected in the study.According to the random number table method, the patients were divided into control group and treatment group, with 45 cases in each group.The control group was treated with ethinylestradiol and cycloprogesterone, while the observation group was treated with ethinylestradiol and cycloprogesterone combined with metformin.The clinical effects of two groups were compared, including estradiol (E2), testosterone (T), luteinizing hormone (LH), follicle stimulating estrogen (FSH), insulin resistance index (HOMA-IR), fasting plasma glucose (FPG) before and after treatment.Body mass index (BMI) and fasting insulin (FINS) levels were measured.The ovulation, pregnancy, menstrual recovery and adverse reactions of the two groups were analyzed. Results The total effective rate of the treatment group (97.77%) was significantly higher than that of the control group (77.77%), the difference was statistically significant (χ2=8.389, P=0.003). Before treatment, there were no statistically significant differences in E2, T, LH, FSH between the treatment group and the control group (all P>0.05). After treatment, the levels of E2[(112.90±18.90)pmol/L], T[(1.30±0.78)nmol/L], LH[(8.00±1.50)U/L]and FSH[(1.20±0.39)U/L] in the treatment group were significantly lower than those in the control group[E2(128.90±19.95) pmol/L, T(2.19±1.00) nmol/L, LH(11.65±1.60)U/L and FSH(1.89±0.50)U/L], the differences were statistically significant(t=3.905, 4.707, 10.871, 7.299, all P 0.05). After treatment, the levels of HOMA-IR (2.19±0.50), FPG[(4.30±1.19)mmol/L], BMI[(22.40±1.89)kg/m2], FINS[(15.98±5.00)mU/L] in the treatment group were significantly lower than those in the control group[HOMA-IR(3.90±0.58), FPG (6.09±1.20) mmol/L, BMI (24.69±4.60)kg/m2, FINS (19.00±6.89)mU/L], the differences were statistically significant(t=14.979, 7.105, 3.089, 2.379, all P<0.05). The ovulation, pregnancy and menstrual recovery in the treatment group were significantly higher than those in the control group (χ2=4.121, 4.285, 10.000, all P<0.05). The incidence of adverse reactions in the treatment group (13.33%) was significantly lower than that in the control group (37.77%), the difference was statistically significant between the two groups (χ2=7.066, P=0.007). Conclusion Ethinylestradiol cycloproterone combined with metformin in the treatment of PCOS with insulin resistance has significant clinical efficacy, can alleviate the endocrine metabolic disorders and insulin resistance, it is worthy of clinical application and promotion. Key words: Polycystic ovary syndrome; Insulin resistance; Ethinyl estradiol; Gyproterone; Metformin; Sex hormones; Lipid metabolism

Comparison of clinical efficacy of dydrogesterone and progesterone in the treatment of threatened abortion

Objective To compare the clinical effect of dydrogesterone and progesterone in the treatment of threatened abortion. Methods From March 2015 to May 2017, 50 patients with threatened abortion admitted in the Maternal and Child Health Hospital of Ninghai County were selected and randomly divided into two groups according to the digital table, with 25 cases in each group.The control group was treated with progesterone.The observation group was treated with dydrogesterone.The hormone levels, symptom disappearance time, fetal retention and adverse drug reactions were compared between the two groups. Results One week after treatment, the levels of progesterone, estradiol and human chorionic gonadotropin in the observation group were (1 12.65±6.35)nmol/L, (2 137.99±371.22)pmol/L, (4 261.54±245.55)IU/L, respectively, which in the control group were (101.56±5.32)nmol/L, (1 782.12±350.71)pmol/L, (3 642.26±210.20)IU/L, respectively, the differences between the two groups were statistically significant (t=6.694, 3.484, 9.581, all P 0.05). Conclusion The effect of dydrogesterone in the treatment of threatened abortion is better than progesterone. Key words: Abortion, threatened; Progesterone; Dydrogesterone; Receptors, progesterone; Estradiol; Chorionic gonadotropin; Drug therapy; Comparative study

Abstract P400: The Association Between Cross-sex Hormone Therapy and Cardiovascular Disease Risk Factors Among Male-to-female Transgender Persons in Chiang Mai, Thailand

Background: Cross-sex hormone therapy (CSHT) is prescribed to transition secondary sexual characteristics among individuals undergoing male-to-female (MtF) transitions (age range 18-41, mean age=24). Limited data exist to inform the cardiovascular disease (CVD) risk factor profile associated with CSHT. We investigated the relationship between CSHT and cardiovascular risk factors in MtF transgender persons and hypothesize that CSHT will be associated with adverse CVD risk factor profiles. Methods: A cross-sectional study was conducted from October 1 st , 2018 to November 30 th , 2018 in 100 MtF transgender people not receiving CSHT vs. 100 receiving CSHT. CSHT use was defined by self-report use of up to 23 medications. Serum testosterone and 17-beta estradiol were assessed to validate CSHT use. Systolic and diastolic blood pressure was measured. Lipid profiles, fasting plasma glucose (FPG), C-reactive protein, cardiac troponin I and pro b-type natriuretic peptide (proBNP) were assessed from fasting blood. Non-invasive arterial examinations included: carotid intima-media thickness (CIMT), ankle-brachial index (ABI), cardio-ankle vascular index (CAVI), and pulse wave velocity (PWV). Multivariable linear regression models, regressed CVD risk factors on CSHT status. Among the subgroup of CSHT users, we assessed the relationship between duration of use and CVD risk factors. Multivariable models included age, gender, education, income, drinking, smoking, exercise, and BMI. Results: Participant mean age was 24±0.38 years and did not differ by CSHT use. Mean±SE values of testosterone were in the CSHT vs. control group were 4.8±0.3 vs. 5.8±0.3 ng/ml, p=0.06 and 17-beta estradiol levels were 45.6±14.9 vs. 34.7±14.8, p=0.7). CIMT was modestly lower among CSHT vs. controls (0.35±0.01 vs. 0.38±0.01, p=0.09). The average duration of CSHT use was 6.65±0.522 years. Among CSHT users, for every 1-year increase in duration of CSHT use total cholesterol decreased by -2.360 ± 1.096, p=0.0341 mg/dL, LDL-cholesterol decreased by -3.076 ± 1.182, p=0.0109 mg/dL, ABI decreased by -0.006 ± 0.002, p=0.0087 while FPG increased by 2.558 ± 0.899 mg/dL, p=0.0055. Conclusion: Among MtF transgender persons, using CSHT was not associated with increased CVD risk factors levels.

4-Hexylresorcinol Exhibits Different Characteristics to Estrogen

4-Hexylresorcinol (4HR) has been used as a food additive and antiseptic. The aim of this study was to evaluate whether the application of 4HR in breast cancer cells and ovariectomized rats showed estrogen-like effects. MCF-7 and SK-BR-3 cells were treated by solvent, 1–100 µM bisphenol-A (BPA), or 1–100 µM 4HR, respectively. 3-(4, 5-Dimethylthiazole-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) assay and Western blot for extracellular signal-regulated kinase-1/-2 (ERK1/2), phosphorylated ERK1/2 (p-ERK1/2), estrogen receptor-α (ERα), and ERβ were done. As an in vivo study, ovariectomized rats (n = 15) received solvent, 125 mg/kg of 4HR, or 10 µg of 17-β estradiol via daily subcutaneous injection for 7 days. Blood samples were obtained for evaluation of prolactin levels. Pituitary glands and uteruses were biopsied for histological evaluation and Western blot analysis. Compared with the control group, the application of 4HR decreased the proliferation of MCF-7 and SK-BR-3 cells, while the application of BPA increased (p &lt; 0.05). The application of BPA increased the expression of ERα, ERβ, and p-ERK1/2, but 4HR did not change the expression of ERα, ERβ, or p-ERK1/2 in MCF-7 cells. In an animal model, the 4HR group showed similar levels of ERα, ERβ, and prolactin expression in the pituitary gland compared to the solvent only group, while the estradiol group showed higher levels. Serum prolactin levels were similar between the 4HR and solvent only groups. Taken together, 1–100 µM 4HR did not show BPA-like behavior in MCF-7 cells, and 125 mg/kg of 4HR daily subcutaneous injection for 7 days did not demonstrate estradiol-like effects in ovariectomized rats. Collectively, 4HR has no estrogen-like effects on both ERα-positive cells and estrogen-deficient rat models.

Clinical observation and study of hormone replacement therapy on postoperative cervical squamous cell carcinoma

Objective To observe the clinical effect of hormone replacement therapy(HRT) on patients with cervical squamous cell carcinoma after operation and its effect on sex hormone, blood fat, bone content and tumor markers, and to evaluate the clinical significance of HRT therapy. Methods From January 2016 to January 2019, a total of 100 patients with cervical squamous cell carcinoma admitted to Zhuhai Maternal and Child Health Hospital were analyzed retrospectively.After surgical treatment, 50 patients were willing to receive hormone replacement therapy as the experimental group and 50 patients received routine treatment as the control group, Kupperma score was used to evaluate the severity of perimenopausal symptoms, and the serum levels of sex hormones, blood fat, bone content and tumor markers in the two groups before and after treatment were detected and the occurrence of adverse reactions were evaluated. Results Before treatment, there was no significant difference in kupperma score, estradiol, follicle-stimulating hormone, luteinizing hormone level, lipid level, bone content and tumor marker level between the two groups(all P>0.05). After HRT treatment, kupperma score in the experimental group was significantly lower than that before treatment((11.47±5.12) vs.(20.46±7.52), t=7.262). In the detection of sex hormones, the levels of estradiol in the experimental group was significantly increased((39.26±7.43) ng/L vs.(13.78±7.52) ng/L, t=12.345), and the levels of follicle stimulating hormone((34.25±7.61) U/L vs.(62.18±19.12) U/L, t=8.245)and luteinizing hormone((20.31±6.25) U/L vs.(35.08±6.27) U/L, t=5.452, P=0.004) in the experimental group were significantly decreased.In the detection of blood lipid level, bone content and tumor markers, high-density lipoprotein((1.62±0.33) mmol/L vs(1.34±0.26) mmol/L, t=4.592, P=0.008) and alkaline phosphatase levels((66.21±25.75) U/L vs.(46.88±9.06) U/L, t=5.912, P=0.001)was significantly increased, the low density lipoprotein((2.78±0.43) mmol/L vs.(2.87±0.78) mmol/L, t=4.265, P=0.012)and total cholestenone((4.02±0.45) mmol/L vs.(4.23±0.91) mmol/L, t=5.761, P=0.002) levels were significantly decreased in the experimental group, with statistically significant differences(all P 0.05); there was no significant difference in the above indicators between the experimental group and the control group before and after treatment (P> 0.05); there was no significant increase in adverse reactions such as breast swelling and pain (3 cases), body mass increase (2 cases), bleeding (1 case), body pain (4 cases), vomiting (4 cases), etc. between the experimental group and the control group (2, 1, 2, 3, 1 case) There was no statistical significance (P> 0.05). Conclusion HRT treatment of cervical squamous cell carcinoma patients after surgery can significantly improve the peri menstrual syndrome caused by low estrogen level, and did not significantly increase the risk of recurrence of cervical cancer patients and adverse reactions. Key words: Hormone replacement therapy; Cervical squamous cell carcinoma; Clinical observation

Ultrasmall Au(0) Inserted Hollow PCN-222 MOF for The High-Sensitive Detection of Estradiol.

Here, the effect of morphology on the electrocatalytic activity of metal-organic framework (MOF) was investigated. Post synthetic gold (Au) insertion was done into hollow PCN-222 (HPCN-222) and solid PCN-222 (SPCN-222) frameworks by a simple hydrothermal method. The crystalline nature, chemical composition, and morphologies of the synthesized MOFs were characterized by PXRD, XPS, and TEM. Electrochemical characterizations were done by cyclic voltammetry and electrochemical impedance spectroscopy. The excellent electrocatalytic activity of highly small-sized Au(0) with the enhancement of electrical conductivity through a hopping mechanism combined with a hollow structure and high surface area in the HPCN-222 MOF hugely alters the electrochemical properties. Overall, a better electrocatalytic surface area, charge transfer coefficient, and catalytic activity were generated at the modified electrode. The hollow structure MOF showed better electrocatalytic activity than solid structured MOF. The AuHPCN-222 modified glassy carbon electrode (AuHPCN-222/GCE) electrochemical sensor was employed for the analytical analysis of estradiol (ED) in an optimum experimental condition. AuHPCN-222 showed selective and better electrocatalytic activity toward ED than GCE. The differential pulse voltammetric measurements by the fabricated sensor showed two linear determination ranges for ED; those are 0.01-1 and 1-220 μM. The measured limit of detection (m-LOD) was found to be 0.5 nM with the highest sensitivity of 2.2 μA μM cm-2. The analytical performances for the detection of ED in human urine and serum samples were done with reasonable recoveries (98%-103%) using the standard addition method. Also, the sensor showed better reproducibility and repeatability with stability.