The objectives of study were to explore the effects of exogenous methyltestosterone( MT) on the growth and gonadal development of overwintering Whitmania pigra. Before overwintering,0. 1,1. 0,10. 0,100. 0,150. 0 μg·L^-1 of MT were added to the aquaculture water for 6 weeks. The changes of growth performance,gonad index,endogenous steroid hormones level and internal quality were measured after hibernate for 60 days. Then the tissue slice technique was used to observe the spermary( ovary) of Wh. pigra.The results showed that the body weight,survival rate and gonadal index increased first and then decreased with the increase of exogenous MT concentration; the male gland index was found the highest at the concentration of MT 10. 0 μg·L^-1 and the female gland index was the highest at the concentration of MT 1. 0 μg·L^-1. The survival rate of Wh. pigra peaked at the concentration of MT 10. 0 μg·L^-1.The weight reaches a peak at a concentration of MT 100. 0 μg·L^-1( P<0. 05). The number of primary spermatocytes in the testis was negatively correlated with the concentration of exogenous MT. The number of secondary spermatocytes and sperm cells increased first and then decreased. The concentration of secondary spermatocytes was the highest when the concentration of MT was 100. 0 μg·L^-1.The number and volume of oocytes in the ovary and the yolk granules increased first and then decreased with the increase of exogenous MT concentration,and the highest was observed at the MT concentration of 100. 0 μg·L^-1. The endogenous steroid hormone of Wh.pigra increased first and then decreased with the increase of exogenous MT concentration. The concentration of androgen and progesterone was the highest in MT 100. 0 μg·L^-1 treatment( P<0. 05),and the concentration of estrogen was found the highest in MT 10. 0 μg·L^-1 treatment( P<0. 05). After adding exogenous MT,Wh. pigra moisture content,acid-insoluble ash content,p H and anti-thrombin activity met the quality criterion of medicinal Wh. pigra in Chinese Pharmacopoeia( 2015 edition). In conclusion,the short-term addition of 1. 0-100. 0 μg·L^-1 exogenous MT before hibernation can promote the growth,the development of sperm cells and the antithrombin activity of Wh. pigra.
Animals ; Estrogens ; Female ; Gonads ; Leeches/physiology* ; Male ; Methyltestosterone ; Ovary ; Progesterone

BACKGROUNDPolycystic ovary syndrome (PCOS) is the commonest endocrinopathy in women of reproductive age. The patients often develop insulin resistance (IR) or hyperinsulinemia despite manifesting anovulation and signs of hyperandrogenism. The cause and effect relationship of hyperinsulinemia and hyperandrogenemia (HA) is still debated. Micro-ribonucleic acids (miRNAs) have recently been shown to play a role in regulation of ovarian function. Our current study focused on the altered expression of miRNAs with PCOS.

METHODSOvarian theca interna tissues were obtained from 10 PCOS patients and 8 controls that were non-PCOS and had normal insulin sensitivity undergoing laparoscopy and/or ovarian wedge resection. Total RNA of all samples was extracted. We studied the repertoire of miRNAs in both PCOS and non-PCOS women by microarray hybridization. Bioinformatic analysis was performed for predicting targets of the differentially expressed miRNAs. Furthermore, selected miRNAs were validated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR).

RESULTSA total of 27 miRNAs were differentially expressed in PCOS patients with respect to the controls in our discovery evaluationand two (miR-92a and miR-92b) of them were significantly downregulated in PCOS women in followed validation (P < 0.05). Targets prediction revealed that miR-92a targeted both GATA family of zinc finger transcription factor GATA-binding factor 6 (GATA6) and insulin receptor substrate proteins 2 (IRS-2).

CONCLUSIONSMiRNAs are differentially expressed between PCOS patients and controls. We identified and validated two miRNAs-miR-92a and miR-92b. They are significantly downregulated and may be involved in the pathogenesis of PCOS.

Adult ; Female ; Humans ; Hyperandrogenism ; genetics ; Insulin Resistance ; genetics ; physiology ; Male ; MicroRNAs ; genetics ; Ovary ; metabolism ; Polycystic Ovary Syndrome ; genetics

The aim of this study is to evaluate pituitary-ovarian function at different postpartum periods during the lactational amenorrhea in order to understand the mechanism by which puerperal lactation is associated with a protracted period of amenorrhea and natural infertility. Ninety four lactating women and 119 lactating women with menstruation, aged between 21 and 38 years, volunteered for this study. The pituitary was relatively insensitive to LH-RH during the first 3 weeks following delivery. The recovery of FSH responsiveness to LH-RH occurred earlier than that of LH. Normal FSH response resumed in the 2nd week while the LH response, although not normal, started at the 3rd week postpartum. Pituitary responsiveness after the 5th week postpartum was similar to that occurring in normally menstruating women, except that FSH response was exagerated. Serum prolactin levels were elevated above 160 ng/ml until the 5th week postpartum and decreased to 84.2 ng/ml in the 6th week postpartum. It appears that at least one reason for anovulation during the first four weeks following delivery is the relative insensitivity of the pituitary to hypothalamic stimulation. Prolactin does not seem to modulate pituitary responsiveness to LH-RH. In order to clarify hormonal profiles during the lactational amenorrhea beyond the 5th week puerperium, serum levels of LH, FSH, prolactin, estradiol and progesterone were determined during different postpartum periods. Serum FSH and LH levels during 1-10 months postpartum were similar to basal levels seen during the normal menstrual cycle. Serum estradiol concentrations throughout 1-10 months postpartum, however, were significantly decreased as compared with the levels during the follicular phase of the normal menstrual cycle. Serum prolactin levels were elevated throughout 1-10 months postpartum in lactating amenorrhic women but decreased as the postpartum period lengthened. As compared with lactating amenorrhic women, lactating women with resumed menstruation showed a decrease in prolactin levels from 89.20 ng/ml to 51.39 ng/ml at 1-3 months, from 75.08 ng/ml to 49.99 ng/ml at 4-6 months, and from 54.73 ng/ml to 28.74ng/ml at 7-10 months postpartum. These results suggest that the apparent anovulation seen beyond 5th week postpartum during lactation was not due to pituitary insensitivity to LH-RH. Rather, prolactindependent mechanism interfering with cyclic activity may be operative during long term lactation.
Amenorrhea/etiology* ; Female ; Gonadorelin/pharmacology ; Gonadotropins, Pituitary/secretion ; Human ; Lactation* ; Ovary/physiology* ; Pituitary Gland/physiology* ; Pregnancy ; Prolactin/physiology ; Puerperium*

Melatonin (N-acetyl-5-methoxytryptamine, MT) is a hormone synthesized and secreted by the pineal gland. Recent studies show that melatonin plays an essential role in the pathogenesis of many reproductive processes. High-concentration melatonin exists in human preovulatory follicular fluid and melatonin receptors are present in ovarian granulosa cells, which indicates the direct effects of melatonin on ovarian function. Reactive oxygen species are involved in a number of reproductive events, including folliculogenesis, follicular atresia, ovulation, oocyte maturation, and corpus luteum formation. Melatonin and its metabolites, as powerful antioxidants and free radical scavengers, can potentially inhibit premature ovarian failure. Literature published in recent years shows the essential roles of melatonin in improving human ovarian function and oocyte quality as well as in the management of infertility. Researches on the action mechanisms of melatonin may provide a theoretical basis for the prevention and treatment of some clinical diseases.
Female ; Granulosa Cells ; metabolism ; physiology ; Humans ; Melatonin ; metabolism ; physiology ; Ovarian Follicle ; growth & development ; metabolism ; Ovary ; physiology ; Reactive Oxygen Species ; metabolism

Anti-Müllerian hormone (AMH) is a member of the transforming growth factor (TGF)-beta superfamily and mainly expressed by the granulosa cells of ovarian follicles. In women AMH is only expressed in ovarian follicles and therefore can be used for the evaluation of the ovarian reserve function and the prediction of ovary ageing and ovarian response during in vitro fertilization (IVF) treatment. This article summarizes the clinical application of AMH, especially in evaluating ovarian reserve functions.
Anti-Mullerian Hormone ; blood ; Biomarkers ; blood ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; blood ; Humans ; Ovarian Follicle ; physiology ; Ovarian Hyperstimulation Syndrome ; prevention & control ; Ovary ; physiology ; Polycystic Ovary Syndrome ; blood

Ovarian cryopreservation can save a large number of germ cells during the cryogenic process, and can restore ovarian endocrine function and ovulation potential after thawing and transplantation, which is an ideal way to retain fertility for patients with cancer. Many factors influence the effect of ovarian cryopreservation, like cryoprotectant (CPA), frozen carrier, cortical block size and freezing procedure. An efficient and standard transplantation procedure is needed to develop for further clinical application and scientific research. In order to optimize this technology, we analyzed different factors to improve the recovery of ovarian function after freezing during ovarian cryopreservation.
Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Female ; Humans ; Ovary ; anatomy & histology ; physiology

OBJECTIVETo observe the protective effect of Oviductus Ranae (OR) capsules on the reproductive organs in an aged mouse model established by D-galactose injection.

METHODSForty-eight female Kunming mice were randomly divided into 4 equal groups, namely the high- and low-dose OR groups, diethylstilbestrol (DT) group, and model group. The mice received subcutaneous injection of D-galactose for 6 weeks to establish aging models. Another 12 mice were injected daily with normal saline (NS) to serve as the normal control group. From the third week of the experiment, the mice were given oral OR at low or high doses (in the OR groups) or vegetable oil (in the model or control groups) till the sixth week. In the last two weeks, the vaginal smears were obtained from the mice for evaluating the changes of the vaginal keratinocytes and counting the days of estrus. After completion of drug administration, all the mice were sacrificed and the serum content of estradiol (E(2)) was detected by radioimmunoassay, with the ovarian and uterine indices determined. The ovarian and uterine pathologies were observed using HE staining, and SOD and MDA activities in the ovary and uterus were also assessed.

RESULTSOR obviously increased E(2) level and the ovarian and uterine indices in the aged mice, also alleviating the pathological change of the ovary and uterus. OR substantially depressed MDA content and enhanced SOD activity in the ovary and uterus.

CONCLUSIONOR has definite antioxidative effects and ameliorates the degenerative changes of the reproductive organs in mouse models of aging.

Aging ; Animals ; Capsules ; Estradiol ; blood ; Female ; Materia Medica ; pharmacology ; Mice ; Ovary ; drug effects ; physiology ; Random Allocation ; Uterus ; drug effects ; physiology

OBJECTIVETo investigate the influence of premenopausal hysterectomy on the function of the conserved ovaries.

METHODSA retrospective survey was conducted by sending questionnaire to 521 cases of hysterectomized women (with the conservation one or two ovaries) for benign gynecological diseases and 1,600 cases of normal controls (with contact uterus and bilateral ovaries). Age, type of operation, diagnosis and the time of the onset of the menopausal syndrome were asked in the questionnaire.

RESULTSThe mean age of the onset of the hot flush (293 cases) in the hysterectomized group was (44.64 +/- 4.31) years, which was significantly lower than that of the normal group [(46.87 +/- 4.22) years, 197 cases] (P < 0.01). The mean age of the above 293 women undergoing hysterectomized was (42.84 +/- 4.37) years. The range of the operation was positively correlated with the time of the onset of the menopause syndrome. So there was only (1.99 +/- 2.40)-year interval between the hysterectomy and the onset of the hot flush.

CONCLUSIONHysterectomy with the conservation of bilateral/unilateral ovaries may have some certain influence on ovarian function.

Adult ; Female ; Hot Flashes ; etiology ; Humans ; Hysterectomy ; Middle Aged ; Ovary ; physiology ; Premenopause ; physiology ; Retrospective Studies ; Surveys and Questionnaires

OBJECTIVETo assess the recovery of the reproductive endocrine function in rats following orthotopic transplantation of fetal ovarian allograft.

METHODSNinety female SD rats (50-60 days old) were randomized into graft recipient group (n=50), positive control group (n=20), and negative control group (n=20) to receive orthotopic transplantation of fetal (17-19 gestational days) ovaries following bilateral oophorectomy, sham abdominal surgery, and bilateral oophorectomy, respectively. At 45 days after the surgeries, serum estradiol and progesterone levels were measured and the ovaries were removed for evaluation of the ovarian volume and follicle development.

RESULTSOn day 45 after the operations, the estradiol or progesterone levels showed no significant difference between the recipient group and positive control group (P>0.05), but both were significantly lowered in the negative control group (P<0.05). The ovarian volume was comparable between the recipient group and positive control group (P>0.05), and optical microscopy showed follicles in different stages of development and formation of corpus luteum in the ovaries in both groups.

CONCLUSIONFetal rat ovary allografts can develop into functional ovaries capable of ovulation to restore the reproductive endocrine function of recipient female rats.

Animals ; Estradiol ; blood ; Female ; Fetus ; Ovariectomy ; Ovary ; physiology ; transplantation ; Ovulation ; physiology ; Pregnancy ; Progesterone ; blood ; Rats ; Rats, Sprague-Dawley ; Transplantation, Homologous

Supplementation of beta-mercaptoethanol (beta-ME) in in vitro maturation (IVM) medium was shown to improve embryo development and quality in several species. Epidermal growth factor (EGF) was also shown to improve IVM of human oocyte and embryo development after in vitro fertilization (IVF). The effect of these two compounds were suggested to be mediated through the synthesis of glutathione (GSH) which is known to play an important role in protecting the cell or embryos from oxidative damage. Thus, it is suggested that supplementation of canine IVM medium with beta-ME or EGF may be of benefit due to its positive role in IVM of various mammalian oocytes and embryo development, including cattle, pigs, rodents and humans. This study investigates the effect of ovarian estrus stage on canine oocyte quality and supplementation of medium with beta-ME or EGF on IVM of canine oocytes. As results, a significantly higher percentage of oocytes progressed to metaphase II (MII) stage in 50 or 100 microM of beta-ME supplemented oocytes collected from the follicular stage. The maturation rate to metaphase I (MI) stage was also significantly higher in oocytes collected from follicular stage and cultured with 25 or 100 microM compared to other experimental groups. After IVM culture, oocytes recovered from dogs with the follicular stage and matured in TCM-199 supplemented with 20 ng/ml EGF yielded better oocyte maturation to MII phase compared to other groups. Taken together, supplementation of beta-ME (50 or 100 microM) or EGF (20 ng/ml) improved IVM of canine oocytes to MII stage.
Animals ; Benzimidazoles/chemistry ; Dogs/*physiology ; Epidermal Growth Factor/*pharmacology ; Estrus/*physiology ; Female ; Fluorescent Dyes/chemistry ; Meiosis/drug effects/physiology ; Mercaptoethanol/*pharmacology ; Microscopy, Ultraviolet/veterinary ; Oocytes/drug effects/growth&development/*physiology ; Ovary/drug effects/*physiology