Estradiol Research Articles

Reproductive hormones influence zinc homeostasis in the bovine cumulus-oocyte complex: Impact on intracellular zinc concentration and transporters gene expression

Zinc (Zn) is a vital trace element for the body and its bioavailability influences numerous reproductive events. However, the mechanisms that regulate Zn homeostasis in the cumulus-oocyte complex (COC) are yet to be elucidated. The aim of this study was to investigate the role of estradiol 17-beta (E2), FSH and LH in Zn homeostasis regulation in bovine COC matured in vitro and Zn transporters gene expression. For this purpose, intracellular Zn levels in oocytes and cumulus cells (CC) were assessed using a Zn-specific fluorescent indicator. In addition, gene expression and sequencing of six Zn transporters (Slc39a6, Slc39a8, Slc39a14, Slc30a3, Slc30a7 and Slc30a9) were assessed. Our results demonstrated that the simultaneous presence of E2, FSH, and LH during oocyte maturation altered intracellular zinc levels and transporters expression in both oocytes and CC. Transporter’s gene expression was different in oocytes and CC, possibly due to cell-specific changes in Zn levels during maturation. The interaction effects of Zn with hormonal treatments influenced the results. This study emphasizes that Slc39a6 is highly sensitive to hormone induction. Overall, the hormonal modulation of Zn homeostasis in the COC was evidenced. Also, a preponderant role of FSH as a modulator of Zn intracellular levels and transporter gene expression is suggested.

The etiology and therapeutic strategies of premature ovarian insufficiency in children and adolescents

Premature ovarian insufficiency(POI), previously known as premature ovarian failure(POF), is defined as the occurrence of premature ovarian failure in women under the age of 40 who may present with delayed puberty, primary or secondary(for 4 months or more)amenorrhea with two times and more of abnormal serum hormonal levels(low estradiol and raised gonadotrophins). And POI is not common in children and adolescents.It is generally believed that the etiologies of POI may be genetic, immunological, iatrogenic, etc.Hormone replacement therapy(HRT)is the primary treatment of POI in children and adolescents, and evaluation of therapeutic strategies to POI including cardiovascular, skeletal and emotional health is needed as well.Recently, the incidence of POI in children and adolescents has increased, therefore, early diagnosis and active treatment are urgent and necessary.Thus, this review will mainly be focused on the etiology and treatment of POI in children and adolescents. Key words: Children; Adolescents; Premature ovarian insufficiency; Etiology; Hormone replacement therapy

Ovariectomized rat model of osteoporosis: a practical guide.

Osteoporosis affects about 200 million people worldwide and is a silent disease until a fracture occurs. Management of osteoporosis is still a challenge that warrants further studies for establishing new prevention strategies and more effective treatment modalities. For this purpose, animal models of osteoporosis are appropriate tools, of which the ovariectomized rat model is the most commonly used. The aim of this study is to provide a 4-step guideline for inducing a rat model of osteoporosis by ovariectomy (OVX): (1) selection of the rat strain, (2) choosing the appropriate age of rats at the time of OVX, (3) selection of an appropriate surgical method and verification of OVX, and (4) evaluation of OVX-induced osteoporosis. This review of literature shows that (i) Sprague-Dawley and Wistar rats are the most common strains used, both responding similarly to OVX; (ii) six months of age appears to be the best time for inducing OVX; (iii) dorsolateral skin incision is an appropriate choice for initiating OVX; and (iv) the success of OVX can be verified 1-3 weeks after surgery, following cessation of the regular estrus cycles, decreased estradiol, progesterone, and uterine weight as well as increased LH and FSH levels. Current data shows that the responses of trabecular bones of proximal tibia, lumbar vertebrae and femur to OVX are similar to those in humans; however, for short-term studies, proximal tibia is recommended. Osteoporosis in rats is verified by lower bone mineral density and lower trabecular number and thickness as well as higher trabecular separation, changes that are observed at 14, 30, and 60 days post-OVX in proximal tibia, lumbar vertebrae and femur, respectively.

Effects of vitamin E on nicotine-induced lipid peroxidation in rat granulosa cells: Folliculogenesis

In this study, we investigated the mechanism of oxidative damage induced by nicotine and the efficacy of vitamin E, an integral component of cellular membranes, against the damage in follicular/granulosa cells of rat ovaries. The animals were randomly divided into 4 groups; control, nicotine, nicotine + vitaminE, vitamin E (n = 8, per each group). Nicotine and vitamin E were administrated intraperitoneally 1 mg/kg/day and 200 mg/kg/day, respectively, once daily for 2 weeks. Nicotine increased lipid peroxide levels such as lipid peroxide (LPO) and malondialdehyde (MDA) in serum, 4-hydroxynonenal (4-HNE) in granulosa cells and apoptotic granulosa cells in the ovary. Positive correlation occurred between the findings of LPO markers and TUNEL labeling. Level of 17-β estradiol (E2), number of follicles and granulosa cell proliferation decreased with nicotine treatment and negatively correlated with LPO levels and apoptosis in granulosa cells. Ultrastructural study of nicotine treated rat ovaries showed mitochondrial damage and autophagosomes in the granulosa cells. The administration of nicotine and vitamin E together, revealed an increase in E2 level, granulosa cell proliferation and the number of healthy follicles associated with decrease in LPO, MDA, 4-HNE levels and TUNEL reactivity in a manner correlated with each other, compared to the nicotine group. Vitamin E showed to alleviate mitochondrial damage and decrease the number of autophagosomes in granulosa cells. These results suggest that lipid peroxidation may be one of the nicotine’ damage mechanisms on folliculogenesis and vitamin E may prevent nicotine-induced follicular damage through reducing lipid peroxidation level in granulosa cells.

The association between vitamin D/25(OH)D and reproductive hormone in young women with recurrent aphthous stomatitis: An observational study

Aim: To investigate the association between vitamin D/25-hydroxyvitamin D [25(OH)D] level and reproductive hormone in women with recurrent aphthous stomatitis (RAS). The reproductive hormonal change suggested contributing to development of RAS in women. Studies have also shown that vitamin D deficiency may contribute to the development of RAS. So far, there is no study that reveals the association between vitamin D/25(OH)D and reproductive hormone in RAS. Materials and Methods: A cross-sectional study was performed on 38 female patients with RAS without any systemic history, any habit, or on any medications. All subjects with RAS underwent intraoral examination and hormonal level assessment to examine reproductive hormones (follicle-stimulating hormone [FSH], luteinizing hormone [LH], estradiol, and progesterone), cortisol, and vitamin D/25(OH)D. The demographic characteristics of oral ulcers, predisposing factors, and pain (visual analog scale [VAS]) scores of RAS were also obtained from all participants. The characteristics of subjects, severity of RAS, FSH, LH, estradiol, progesterone, cortisol, and vitamin D/25(OH)D were presented descriptively. Results: All subjects had mean value of estradiol (93.99 ± 81.34 pg/mL), progesterone (0.73 [0.44 to 3.73 ng/mL]), FSH (4.24 ± 1.90 mIU/mL), LH (8.12 ± 5.76 mIU/mL), cortisol (8.13 ± 2.70 µg/mL), and had low mean value of serum vitamin D/25(OH)D (10.88 ± 3.21 ng/mL) which was categorized with vitamin D deficiency. The result of Pearson correlation test showed a significant positive correlation between mean LH and vitamin D/25(OH)D (P < 0.05). Conclusion: All subjects with RAS have vitamin D deficiency and vitamin D is correlated with LH.

Effect of laparoscopic myomectomy on serum sex hormones, inflammatory factors and immune function in patients with uterine fibroids

Objective To investigate the effect of laparoscopic myomectomy on serum sex hormones, inflammatory factors and immune function in patients with uterine fibroids. Methods From January 2014 to May 2017, 100 patients with uterine fibroids who were treated in the People's Hospital of Taigu County were selected.The patients were divided into control group and observation group according to the random number table method, with 50 cases in each group.The control group was treated with conventional myomectomy, while the observation group was treated with laparoscopic myomectomy.The serum levels of follicle stimulating hormone(FSH), luteinizing hormone(LH), estradiol(E2), interleukin 6(IL-6), interleukin 10(IL-10), tumor necrosis factor-α(TNF-α) and immune function were compared before and after operation. Results The serum levels of FSH, LH, IL-10, CD3+ , CD4+ , CD8+ in the observation group after operation were (7.81±2.29)IU/L, (10.52±2.73)IU/L, (12.64±2.18)ng/L, (52.67±4.87)%, (31.56±3.82)%, (22.55±2.16)%, respectively, which were higher than those in the control group[(7.36±2.11)IU/L, (9.86±2.48)IU/L, (8.64±2.03)ng/L, (46.32±4.64)%, (27.62±3.78)%, (16.87±2.14)%], the serum levels of E2, IL-6, TNF-α in the observation group after operation were (65.56±5.45)ng/L, (13.67±3.56)ng/L, (23.58±4.05)ng/L, which were lower than those in the control group[(70.64±5.81)ng/L, (30.57±4.11)ng/L, (27.56±4.21)ng/L], the differences were statistically significant(t=2.542, 2.589, 9.495, 6.675, 5.184, 13.209, 4.509, 21.977, 4.817, all P<0.05). Conclusion Compared with the traditional laparotomy, laparoscopic myomectomy can improve the levels of FSH, LH and E2 in patients with uterine fibroids, relieve postoperative inflammatory response, and improve the immune function of patients. Key words: Uterine Neoplasms; Laparoscopy; Myomectomy; Follicle stimulating hormone; Luteinizing hormone; Estradiol; Interleukin-6; Interleukin-10; Tumor necrosis factor-alpha; Immune function

Evaluation of osteocalcin and reproductive hormones in men with type 2 diabetes in Misan province/Iraq

Background: Diabetes mellitus (DM) is a group of metabolic disorders in which there are high levels of blood sugar over a prolonged period. DM can be accompanied by numerous complications, including different organ systems for instance skeleton and reproductive system. Objectives: The aim of the present study was to determine the levels of osteocalcin (OC) and reproductive hormones in men with type 2 DM (T2DM) of Misan province from December 2018 to May 2019. Materials and Methods: The study population consists of 110 men aged 35–49 years of whom 66 type 2 diabetic and 44 healthy men; patients with T2DM were divided based on the age into three groups: the first (35–39) years, second (40–44) years, and third (45–49) years; patients with T2DM were also divided based on the body mass index into four groups: normal weight (18.5–24.9), overweight (25–29.9), obesity class I (30–34.9), and obesity class II (35–39.9) groups. Results: According to the age, the mean values of fasting blood glucose and glycosylated hemoglobin A1c (HbA1c) were significantly higher in diabetic subjects when compared with the control. OC increases significantly in the second and third groups as compared to control. Luteinizing hormone (LH) of the patients in the third group significantly increases as compared to the first group. Patients of the third group increase significantly compared to the patients in the first group. Follicle-stimulating hormone (FSH) means value no different significantly according to the age. The testosterone (TT) mean value in compare between patients group, the third increase significantly in compare with first group. Estradiol (E2) in the first group increases significantly as compared to control. Prolactin (PRL) in the first group decreases significantly compared to control. According to the BMI, the mean value of fasting blood glucose and HbA1c was significantly higher in diabetic subjects when compared to control. OC value of T2DM significantly increases in the group obesity class II and overweight. LH, TT, and FSH were not significantly significant compared with control. E2 value significant different in compare with control of obesity class I and class II. PRL decreased significantly as compared to control in normal weight group; in patients of obesity class I, the E2 value increases significantly as compared to patients of normal weight.Conclusions: This study revealed no relationship between OC and reproductive hormones and no relationship between OC and T2DM.

Protective effect of laparoscopic removal of water and suture hemostasis on ovarian function

Objective To investigate the protective effect of laparoscopic water separation and removal combined with suture hemostasis on ovarian reserve function after bilateral ovarian endometriotic cyst (OEC) stripping. Methods From January 2016 to January 2018, 60 patients with bilateral ovarian endometriosis cystectomy underwent laparoscopic surgery in the Department of Obstetrics and Gynecology, Jiading Hospital of traditional Chinese medicine.According to the random number table method, they were divided into study group (water separation and stripping combined suture hemostasis group) and the control group (direct stripping combined with electrocoagulation hemostasis group), 30 cases in each group.The changes of operation time, hemoglobin level before and after operation and the levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2) and anti Mullerian hormone (AMH) were compared between the two groups. Results There was no significant difference in operation time (47.52 ±10.11) min, hemoglobin decrease (0.55 ±0.26) g/L, hospital stay (6.1 ±0.3) d, control group (48.01 ±10.24) min, hemoglobin decrease (0.56 ±0.25) g/L and hospitalization time (6.2 ±0.4) d before and after operation (t=0.056, 0.964, 0.863, all P>0.05). The levels of FSH, E2, LH and AMH in the study group before operation were (6.15 ±2.31) U/L, (152.41 ±41.40) nmol/L, (5.44 ±1.52) U/L and (2.21 ±0.13) μg/L, respectively.One month after operation, they were(6.21±2.24) U/L, (150.63±40.33) nmol/L, (5.13±1.58) U/L, (2.18±0.16) μg/L, respectively.Three months after operation, they were (6.52±2.41) U/L, (149.57±42.37) nmol/L, (5.30±1.45) U/L, (2.17± 0.15) μg/L, respectively.Six months after operation, they were (6.53±2.44) U/L, (151.36±41.54) nmol/L, (4.98±1.61) U/L, (2.20±0.08) μg/L, respectively.The levels of FSH, E2, LH and AMH in the control group before operation were (6.14±2.21) U/L, (153.31±40.39) nmol/L, (5.51±1.46) U/L, (2.23±0.13) μg/L, respectively.One month after operation, they were (8.11±2.44) U/L, (131.43±41.23) nmol/L, (5.92±1.64) U/L, (1.58±0.14) μg/L, respectively.Three months after operation, they were (8.42±2.35) U/L, (135.67±40.38) nmol/L, (6.12±1.51) U/L, (1.54±0.16) μg/L, respectively.Six months after operation, they were (9.17±2.64) U/L, (133.66±40.44) nmol/L, (6.28±1.74) U/L, (1.51±0.13) μg/L, respectively.There was no significant difference in the levels of FSH, E2, LH and AMH between the two groups (all P>0.05). There was significant difference between the preoperative FSH level and the postoperative 1, 3, 6 months in the control group (all P<0.05); there was significant difference between the preoperative E2 level and the postoperative 1 month in the control group (P<0.05); there was statistical significance between the preoperative AMH level and the postoperative 1, 3 months in the control group (all P<0.05). The levels of FSH, LH, E2 and AMH in the study group were significantly higher than those in the control group (all P<0.05). Conclusion Laparoscopic hydrodissection combined with suture hemostasis does not increase the operation time and the amount of bleeding, and does not affect the ovarian reserve function after operation. Key words: Ovarian endometrioma; Hydrodissection; Suture hemostasis; Ovarian reserve function

Comparison the Effect of Metformin and Clomiphene Citrate on Sirtuin3 gene Expression in the Oocytes of Mice with Polycystic Ovary Syndrome.

Oxidative stress (OS) is a common biological event in polycystic ovarian syndrome (PCOS), causing oocytes to undergo OS-induced changes. Sirtuin3 (Sirt3) has a critical role in oocyte maturation through the modulation of OS. In the current study, we compared the effects of metformin and clomiphene citrate on the expression of the Sirt3 gene in oocytes obtained from the mice, induced by PCOS. The induction of PCOS was performed by the single injection of estradiol valerate. The animals were divided into control, PCOS, metformin (500 mg/Kg), and clomiphene (18 mg/kg) groups. At the end of the experiment, the levels of LH and FSH were determined using the ELISA method. The ovarian tissues were evaluated histologically, and the expression of the Sirt3 gene was analyzed by the Real-time PCR. The induction of PCOS led to an increase in the ratio of LH/FSH elevation, the number of follicle atresia, as well as the presence of hydrated cysts. The results showed that both treatment regimens returned the altered parameters to the baseline values. The gene of Sirt3 was significantly (P < 0.001) reduced in the PCOS group compared to the control. Also, no significant difference was found in the expression of Sirt3 between clomiphene and PCOS group, whereas, in the metformin group, Sirt3 expression had the higher rate of expression in comparison with the PCOS group (P < 0.05). The administration of metformin and clomiphene showed that metformin is capable of preventing the downregulation of the Sirt3 gene in oocytes, collected from PCOS mice.

Эффективность применения гепариноидов у пациенток с дисфункцией эндометрия на фоне недифференцированной дисплазии соединительной ткани

Objective. To study the effectiveness of heparinoid sulodexide in complex therapy of patients with endometrial dysfunction and concomitant undifferentiated connective tissue disease (UCTD). Patients and methods. We examined 88 patients with UCTD and endometrial dysfunction (the «thin endometrium» phenomenon). The therapeutic regimen of 41 patients (group 1а) additionally to cyclical therapy with 17-b estradiol and didrogesterone was supplemented by continuous heparinoid sulodexide for 3 menstrual cycles (MC), 20 patients (group 1b) in addition to cyclical therapy received the heparinoid from the 5th day of MC until the 7th day after ovulation, 27 patients of the reference group (group 2) received only cyclical therapy without heparinoid supplementation. Results. In patients of groups 1а and 1b, restoration of M-mode echo values ≥7mm was obtained in 71–73% of cases, the median endometrial thickness in group 1а increased by 2.8 mm, in group 1b – by 2.25 mm, in group 2 – only by 1.9 mm. The continuous intake of, including during menstruation, was not associated with an increase in blood loss nor changes in red blood counts. As was also found, in patients with UCTD who had algomenorrhoea with ultrasonic signs of varicose veins of the uterus administration of sulodexide resulted in a decrease of the severity of pain syndrome up to its complete disappearance. Conclusion. The treatment regimen for patients with endometrial dysfunction concomitant with UCTD НДСТ with inclusion of continuous heparinoid sulodexide (without a break during menstruation) enhances the effectiveness of therapy, does not involve a higher menstrual blood loss, and also has a positive effect on the severity of algomenorrhoea. Key words: algomenorrhoea, undifferentiated connective tissue disease, sulodexide, chronic endometritis

Xiao-Yao-San Formula Improves Cognitive Ability by Protecting the Hippocampal Neurons in Ovariectomized Rats.

Xiao-Yao-San (XYS) decoction is a traditional Chinese medicine formula. This study aimed to investigate the effect of XYS on cognitive abilities and its underlying mechanism in ovariectomized rats. Female Sprague-Dawley rats were ovariectomized and treated with XYS (3 g/kg or 9 g/kg) by gavage, with subcutaneous injection of 17-β estradiol (E2, 2 μg/kg) as a positive drug control and gavage of 1 ml saline (0.9%) as a placebo control. After 6 weeks of treatment, rats were examined using the Morris water maze test. The estradiol level in the serum and hippocampus was measured by ELISA. Golgi staining was performed to observe neuronal morphology in the hippocampus. Apoptosis of hippocampal cells was observed by TUNEL staining. The protein content of N-methyl-D-aspartate receptor (NMDAR) 2A and 2B in the hippocampal CA1 region was determined by Western blot and immunohistochemistry. Expression of estrogen receptor (ER) and PI3K signaling was detected by Western blot. Compared with the sham group, both learning and memory were impaired in ovariectomized rats. Rats treated with E2 or high-dose XYS showed better learning and memory compared with the saline-treated rats. High-dose XYS significantly reduced escape latency in the spatial acquisition trial; meanwhile, the cross times and duration in the probe quadrant were increased in the spatial probe trial. High-dose XYS promoted the de novo synthesis of E2 content in the hippocampus but had no significant effect on the serum E2 level. Golgi staining indicated that high-dose XYS could increase the branch number and density of dendritic spines in the hippocampal CA1 area. TUNEL staining showed that high-dose XYS alleviated ovariectomy-induced neuronal apoptosis. The expression level of NMDAR2A and NMDAR2B in hippocampal CA1 was upregulated by XYS treatment. The beneficial effect of XYS was through activating ERα-PI3K signaling. In conclusion, high-dose XYS treatment can improve the cognitive abilities of ovariectomized rats by protecting the hippocampal neurons and restoring the hippocampal E2 level.

Greater Muscle Stiffness during Contraction at Menstruation as Measured by Shear-Wave Elastography.

It is important to measure mechanical properties of muscle, since muscle stiffness is an important component of stabilizing or controlling joint stability. The levels of sex hormones especially estrogen vary over the phase of the menstrual cycle and impact the mechanical properties of soft tissue such as muscle, tendon, and ligaments due to the presence of 17-β estradiol receptor in human connective tissues. Recently, shear-wave elastography (SWE), based on ultrasound imaging, has been used as an accurate technique for visualizing and assessing tissue stiffness. The purpose of this study was to compare the muscle stiffness at rest and during contraction condition between the early follicular phase (menstruation) and ovulation in young women, measured using SWE. Thirty-seven young women with regular menstrual cycles completed this study throughout one full menstrual cycle. Stiffness of lower limb muscles such as the rectus femoris, biceps femoris, tibialis anterior, and medial gastrocnemius was measured at resting and during contraction conditions using SWE during menstruation and ovulation. All muscles showed significantly greater stiffness during the menstruation than ovulation when muscles were actively contracted (P < 0.05), whereas no significant differences in muscle stiffness at rest were noted across phase of the menstrual cycle. These significant findings suggest that muscular factors are changed with estradiol fluctuations; muscles are less stiff during ovulation where the levels of estradiol peak when muscles in a contraction condition. As muscle stiffness is an important part of joint stability, these differences should be recognized to prevent the risk of musculoskeletal injuries.

Preparation of ferric oxide-aluminium oxide carbon nanofiber nanocomposites for ultrasound-assisted dispersive magnetic solid phase extraction of 17-beta estradiol in wastewater

Abstract In this work, Fe3O4-Al2O3@CNFs nanocomposite was synthesised and used as a nanosorbent in the ultrasound-assisted dispersive magnetic solid phase extraction (UA-DMSPE) of 17-beta estradiol (E2) in wastewater samples. The quantification of E2 was achieved using high performance liquid chromatography coupled with diode array detector (HPLC-DAD). Various parameters affecting the efficiency of this sample preparation technique were optimised to achieve excellent sensitivity and high recoveries of E2. Response surface methodology was utilised for optimisation of these parameters. Using the optimised conditions, the linear dynamic range was achieved in the range of 0.1–1000 μg L−1 and the correlation coefficient was found to be 0.9981. The preconcentration factor, enrichment factor, limit of detection (LOD) and limit of quantification (LOQ) were 67, 169, 0.025 μg L−1 and 0.083 μg L−1, respectively. The relative standard deviation (%RSD) for the intraday (n = 10) and interday (n = 5 working days) were 1.8% and 3.3%, respectively. The developed UA-DMSPE/HPLC-DAD method was applied for the preconcentration and determination of E2 in wastewater samples. The obtained results indicated that E2 was present in the wastewater samples.

Application value of estradiol in seminal plasma in predicting semen parameters of male infertile patients

Objective To research the correlation between estradiol in seminal plasma and semen parameters of male infertile patients and to explore the value of seminal estradiol in predicting sperm quality. Methods A total of 86 infertility men who visited the reproductive center of our hospital from October to December 2018 were randomly selected and were measured semen parameters according to the WHO laboratory standard. We also tested seminal plasma estradiol by chemiluminescence. The patients were divided into four groups according to the seminal parameters: oligozoospermia (group 1, n=12), asthenospermia (group 2, n=24), oligoasthenozoospermia (group 3, n=24), normozoospermia (group 4, n=26). Results The level of estradiol in seminal plasma in group 1 to 4 were respectively (56.97±10.55) pg/ml, (52.30±8.64) pg/ml, (54.02±11.63) pg/ml, and (45.79±5.87) pg/ml, with statistically significant difference (P<0.01). The correlation analysis showed that the seminal plasma estradiol was negatively correlated with sperm concentration and sperm progressive motility, was not correlated with sperm normal morphology and sperm acrosin activity. The ROC curve of estradiol was drawn for the diagnosis of abnormal sperm. The cut off value of curve was 42.50 pg/ml, the sensitivity was 90.00%, the specificity 69.23%, the area under the curve was 0.737 (P<0.01), and the 95% confidence interval was (0.634, 0.840). Conclusion Seminal plasma E2 more than 42.50 pg/ml may be connected with abnormal sperm counts and vitality. Therefore, seminal plasma estradiol has certain forecastable value, which can be used as evaluation index of the diagnosis and treatment of male infertility patients. Key words: Estradiol; Sperm motility parameters; Sperm morphology; Acrosin

MiRNAs Regulating Insulin Sensitivity Are Dysregulated in Polycystic Ovary Syndrome (PCOS) Ovaries and Are Associated With Markers of Inflammation and Insulin Sensitivity.

Objective: MicroRNAs (miRNAs) are gene expression regulators. Altered miRNA levels are associated with diabetes, insulin resistance, and inflammation. Insulin resistance and inflammation are both features of Polycystic ovary syndrome (PCOS). The aim of this study was first to assess differences in selected miRNAs (miR-146a, miR-155, miR-320, miR-370, miR-486), involved in insulin sensitivity regulation and inflammation, in women with or without PCOS. Second, to investigate relationships among these miRNAs, insulin, High mobility group box 1 (HMGB1), and IL-6 in follicular fluid (FF), serum 17-beta estradiol (E2), and the number of dominant follicles.Methods: Thirty PCOS and thirty-six non-PCOS women undergoing in vitro fertilization were enrolled. RNA from granulosa cells (GC) and FF was extracted and the specific miRNAs were evaluated using qRT-PCR. HMGB1, insulin, and IL-6 in FF, and serum E2 were assayed using specific kits.Results: MiR-146a, miR-155, miR-486 were upregulated and miR-320 and miR-370 were downregulated in GC from the PCOS patients. In FF, miR-146a, miR-155, and miR-486 showed lower levels in PCOS, whereas miR-320 and miR-370 showed an opposite trend but no significant changes were observed. These miRNAs showed relationships with Body Mass Index (BMI), age, E2, number of dominant follicles, insulin, and HMGB1.Conclusion: In conclusion, the miRNAs analyzed showed changes in PCOS ovaries and had relationships with indices of inflammation and insulin sensitivity within the ovary, providing evidence for new regulatory mechanisms.

Polythiophene as highly efficient sorbent for microextraction in packed sorbent for determination of steroids from bovine milk samples

Herein, a method based on microextraction in packed sorbent (MEPS) with polythiophene (PTh) as adsorbent material and HPLC-DAD was developed. PTh was synthesized by oxidative method and used to extract progesterone (PGN), prednisolone (PRE) and estradiol (ESD) from bovine milk samples. The adjusted chromatographic conditions were: C18 column (150 mm × 4.60 mm, 5 µm), mobile phase: methanol: acetonitrile (55: 45, v/v) with flow rate at 0.85 mL min−1 and temperature at 40 °C. The main parameters that affecting the sample preparation were optimized: washing solvent, eluent solvent, sample pH, volume sample, mass of sorbent, and cycles of aspersion/dispersion. The method presented linearity in the linear range between 16 and 1200 ng mL−1 and correlation coefficients (r) ≥ 0.99 for the three analytes with recoveries greater than 85%. The validated method was used successfully for the analyses of bovine milk samples.

The effects of methamphetamine on gonadotropin and androgen, estrogen receptors in male rats

Objective To investigate the effects of methamphetamine on gonadotropin and testosterone receptors in male rats. Methods Acute and chronic exposure models were established by intraperitoneal injection of Methamphetamine (METH). Enzyme linked immunosorbent assay (ELISA) was performed to detect the expression of testosterone (T), estradiol (E2), Follicle stimulating hormone (FSH), luteinizing hormone (LH), progesterone (PROG), prolactin (PRL), structural changes of testicular were observed by hematoxylin and eosin (HE) staining, immunohistochemical staining (IHC) and Western blotting were performed to detect the localization and expression of androgen receptor (AR), estrogen receptor alpha (ER) estrogen receptor β (ERβ) in the testis. Results ELISA results showed that compared with control group, the concentration of T in the acute exposure group was (2.16±0.51) μg/L, which was significantly decreased (t=-2.432, P<0.05). The concentrations of T, E2, PRL and INH B in the chronic exposure group were (0.41±0.37) μg/L, (73.84±33.63) ng/L, (5.86±4.27) μg/L, and (22.52±7.19) ng/L respectively. The T, E2 and INH B in the chronic exposed group was significantly decreased compared to the control group (t=-6.211, -2.853, 2.553, P<0.05). While the PRL was significantly increased (t=2.318, P<0.05). HE staining showed that the spermatogenic cells in the METH exposure group were slightly reduced and disordered. Compared with control group, IHC and Western blotting showed that the protein expression of AR, ERxpond ERβ decreased in the testis. Conclusion The acute and chronic exposure of METH in male rats could lead to gonadotropin disorder and decreased expression of testosterone receptors AR, ERed with the chronic controt the occurrence and development of the sperm. Key words: Methamphetamine; Male reproduction; Gonadal hormone; Hormone receptor

349 Empty body composition and retained energy of Jersey steers using an aggressive implant strategy

Abstract This experiment was designed to quantify the empty body composition of Jersey steers administered an aggressive implant strategy. Jersey steers {n = 30; initial body weight (BW) 183 ± 43 kg} were randomly assigned to one of two implant strategies: negative control (CON), or implanted with Revalor 200 (200 mg trenbalone acetate / 20 mg estradiol 17-β; (REV) every 70 d (d 0, d 70, d 140, d 210, d 280, d 350) during a 420 d feeding period. Steers were harvested on d 421; 6 CON and 6 REV steers were randomly selected for collection of blood, hide, ground viscera, bone, and ground lean and fat to determine empty body composition. Proximate analysis was completed for each sample to determine total body percentages of moisture, crude protein, fat, and ash. Data were analyzed via independent t-test. Percentage empty body moisture (46.48% CON vs 49.69% REV) and empty body protein (15.32% CON vs 17.58% REV) were greater (P &amp;lt; 0.01) in REV cattle. In contrast empty body fat (33.51% CON vs 26.93% REV) was greater (P &amp;lt; 0.01) for CON cattle. Empty body ash did not differ (P &amp;gt; 0.10; 4.69% CON vs 5.80% REV) between treatments. Negative control steers contained a total empty body protein to total empty body fat ratio of 0.44:1 compared to 0.62:1 for REV steers. These data suggest that an aggressive implant strategy alters composition of gain during the finishing of Jersey steers toward increased protein and decreased fat.

Relationship between sex hormones, lipids and visceral fat area in postmenopausal women with type 2 diabetes mellitus

Objective To study the characteristics and correlation of sex hormone levels, blood lipids, and visceral fat area in postmenopausal women with T2DM. Methods A total of 258 cases of postmenopausal T2DM were recruited, including 46 cases of menopause for 1-5 years (group A), 49 cases of menopause for 6-10 years (group B), and 163 cases with menopause more than 10 years (C group). The related clinical information of patients was recorded. Results Compared among the three groups, there were statistical differences in total cholesterol(F=3.287, P=0.039) and testosterone(Chi-Square=8.324, P=0.016). No significant difference in FSH, LH, estradiol, low density lipoprotein, triglyceride, visceral fat area and subcutaneous fat area(P>0.05) among three groups was observed. After correction of confounding factors, FSH was independently positively related with for total cholesterol and low-density lipoprotein respectively (P=0.006, P=0.009). LH was independently positively related with total cholesterol(P=0.003) and low-density lipoprotein respectively(P=0.003). Estradiol was independently negatively related with total cholesterol(P=0.014) and low-density lipoprotein(P=0.020), respectively. Testosterone was correlated independently with visceral fat area(P=0.008); FSH, LH, estradiol, and testosterone were not correlated to triglyceride and subcutaneous fat area(P>0.05). Conclusion (1)In T2DM patients with the increase of postmenopausal time, the total cholesterol increases first, and then decreases; testosterone decreases first, and then increases. (2)In T2DM patients, the total cholesterol and low-density lipoprotein increase with the increase of FSH and LH, increase with the decrease of estradiol, and the area of visceral fat increases with the testosterone together. Key words: Diabetes mellitus, type 2; Postmenopausal women; Sex hormones; Blood lipids; Visceral fat area

Applicability of the OECD 455 in-vitro assay for determination of hERa agonistic activity of isoflavonoids

The Organisation for Economic Co-operation and Development (OECD)-validated transactivation assay using the human estrogen receptor alpha (hERα) Hela9903 cell line is used for activity evaluation of hERα agonists and antagonists. Due to many advantages, this assay is broadly used as an initial screening process. However, response significantly higher from that of 17-β estradiol (E2) was observed with phytoestrogens for concentrations commonly above 1 μM in previous studies. The main aim of this study was thus to ascertain the applicability of OECD protocol 455 for evaluation of estrogenic activity of natural flavonoids, including known phytoestrogens. The estrogenic activities of aglycones as well as of O-methylated and glycosylated flavonoids were evaluated. Supra-maximal luciferase activity was seen for most of the flavonoids tested at concentrations even below 1 μM. hERα-mediated luciferase expression was confirmed with the competition assay specified in OECD protocol 455. However, at concentrations above 1 μM, non-specific interactions were also observed. Instead of EC50 values, which could not be determined for most of the isoflavonoids tested, the concentrations corresponding to 10% (PC10) and 50% (PC50) of the maximum activity of the positive control, E2, were used for quantitative determination of estrogenic activities. Appropriate evaluation of the data obtained with the current OECD protocol 455 validated assay represents a valuable tool for initial screening of natural flavonoids for estrogenic activity.