TJZYF Improves Endometrial Receptivity through Regulating VEGF and PI3K/AKT Signaling Pathway.

The endometrium receptivity was impaired by controlled ovarian hyperstimulation (COH), which would then lead to fertility issues and increased abortion clinically. In the present study, to explore the effectiveness of Tiaojing Zhuyun Formula (TJZYF) in improving endometrial receptivity of COH rats and the possible active ingredients and mechanisms, an approach of network pharmacology was performed and a COH animal model was established. As analyzed, stigmasterol and quercetin may be the active ingredients of TJZYF on improving endometrial receptivity and positive regulation of ion transport, the cytokine-mediated signaling pathway, and endocrine process, and vascular endothelial growth factor receptor signaling pathway may be involved. Eighty female rats were divided into four groups randomly: control, model, TJZYF, and TJZYF+si-VEGFA. COH rat models were constructed by injecting with human menopausal gonadotropin (HMG) and human chorionic gonadotropin (HCG). We found that both endometrial thickness and number of embryo implantations in model were substantially reduced vs. control. The gene and protein expressions of VEGF, PI3K, and p-Akt in the uterus were significantly reduced. TJZYF could increase the endometrial thickness and number of embryo implantations and enhance the expressions of VEGF, PI3K, and p-Akt in the uterus. In the TJZYF+si-VEGFA group, the effect of TJZYF was impaired. Generally, TJZYF could improve the endometrium receptivity and facilitate embryo implantation of COH rats by upregulating VEGF and enhancing the PI3K/Akt signaling pathway.

Elevated basal luteinizing hormone does not impair the outcome of human menopausal gonadotropin and medroxyprogesterone acetate treatment cycles.

The potential effects of high basal luteinizing hormone (LH) levels on human reproduction were controversial. To demonstrate the effects of elevated basal LH levels on the outcome of patients undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles, we performed a retrospective data analysis of 1011 polycystic ovarian syndrome (PCOS) patients treated with human menopausal gonadotropin and medroxyprogesterone acetate (hMG + MPA) protocol at our center between Nov. 2013 and Jun. 2017. PCOS patients with elevated basal LH levels had significantly higher LH exposure during the stimulation period. The group with LH ≥ 10 mIU/mL showed a lower mean total hMG dose used but higher numbers of oocytes retrieved, metaphase II oocytes, embryos and top-quality embryos developed than the groups with lower basal LH levels. Moreover, partial correlation analysis showed that the basal LH level was negatively correlated with the total hMG dose but positively correlated with the numbers of oocytes retrieved, metaphase II oocytes, embryos, and top-quality embryos. There were no significant differences in the rates of oocyte retrieval, fertilization, implantation, clinical pregnancy and miscarriage between the groups based on frozen embryo transfer (FET). We concluded that elevated basal LH level does not impair the final outcome of hMG + MPA-treated IVF/ICSI cycles in PCOS women.

Male reproductive endocrinology: when to replace gonadotropins.

Infertility is generally defined as a couple's inability to conceive after 1 year of unprotected intercourse. When infertile couples seek assistance, a male factor will be identified half of the time. Once the male has been evaluated, there are four main categories to describe his infertility: (1) idiopathic, (2) post-testicular/obstructive, (3) primary-where the Sertoli and/or Leydig cells of the testis fail, and (4) secondary-where there is a problem with the hypothalamus and/or pituitary. The last, hypogonadotropic hypogonadism (HH), accounts for up to 2% of infertile men. HH is either congenital or acquired and usually can be successfully treated by medical intervention. This review will focus on the hypothalamus-pituitary-gonadal axis, specific defects of this coordination center, and potential interventions for improving male-factor fertility.

Outcome of gonadotropin therapy for male infertility due to hypogonadotrophic hypogonadism.

Data on the management of male infertility secondary to hypogonadotrophic hypogonadism (HH) are limited. We report our extensive experience with intramuscular injections of gonadotropins, one of the two methods used for this purpose. Eighty-seven married men (median age, 28 years) with either congenital (47 men) or acquired (40 men) HH were treated for a median of 26 months (range, 6-57) with intramuscular injections of gonadotropins (HCG/HMG) three times weekly for the purpose of achieving fertility. The outcome was assessed by achievement of one or more pregnancies. Of the 151 courses of HCG/HMG treatment administered to 87 patients, 85 courses (56.3%) were successful, resulting in 85 pregnancies (median pregnancy rate 2, range 1-10) in 35 patients (40%) while 52 cases did not achieve pregnancy. Responders had larger pretherapy testicular volume (9 +/- 3.6 cc) compared to non-responders [(5.7 +/- 2.0 cc), P < 0.0001], but there was no difference in age, LH, FSH or testosterone levels or doses of HCG/HMG used. The pregnancy rate was similar in those with congenital (51.4%) and acquired causes (48.6%) of HH (P = 0.83). Only testicular size was predictive of conception (P < 0.001, odds ratio 1.5, 95% CI 1.21-1.92) while age, pretherapy levels of testosterone, LH, FSH and doses of HCG/HMG did not predict the success of pregnancy. Gonadotropins are moderately effective in achieving one to several pregnancies in HH. Only testicular size is predictive of success in achieving pregnancy. There is no difference in success between those with congenital and acquired causes of HH.

Analytical identification of additional impurities in urinary-derived gonadotrophins.

Advances in proteomic technology have enabled contaminant proteins to be identified from complex protein mixtures. The purity of two purified urinary gonadotrophin products, human menopausal gonadotrophin (u-HMG) and human FSH (u-hFSH), was compared with a preparation of recombinant human FSH (r-hFSH). After separation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), western blot analysis showed that the recombinant preparation contained only FSH, whereas the urine-derived preparations exhibited several non-FSH or LH-related bands. These urinary components were further investigated by a proteomic approach using two-dimensional SDS-PAGE followed by mass spectrometric identification. The proteomic approach detected a total of 23 non-gonadotrophin-related proteins, at variable levels in different batches of the urine-derived preparations. Of these, 16 co-purified proteins have not been previously reported to be present in urine-derived gonadotrophins. These results indicate that the process used to purify urinary gonadotrophins may not remove all non-gonadotrophin proteins. By using a comprehensive proteomic approach, it has been shown that the recombinant FSH preparation has greater purity than either of the urine-derived preparations.

Primary Ovarian Insufficiency: X chromosome defects and autoimmunity.

Premature ovarian failure (POF) is a primary ovarian defect characterized by absent menarche or premature depletion of ovarian follicles before the age of 40 years. However, in several instances the distinction between definitive or intermittent POF may be difficult on clinical bases, therefore the more appropriate term Primary Ovarian Insufficiency (POI) has been recently proposed and will be used in this review. POI is a heterogeneous disorder affecting approximately 1% of women <40 years. The most severe forms present with absent pubertal development and primary amenorrhea, whereas forms with post-pubertal onset are characterized by disappearance of menstrual cycles (secondary amenorrhea) associated with a defective folliculogenesis. POI is generally characterized by low levels of gonadal hormones (estrogens and inhibins) and high levels of gonadotropins (LH and FSH) (hypergonadotropic amenorrhea). Heterogeneity of POI is reflected by the variety of possible causes, including autoimmunity, toxics, drugs, as well as genetic defects. Several data indicate that POI has a strong genetic component. In this manuscript we discuss the X chromosome abnormalities that are associated with POI.

[Characterization of DNA-protein cross-links forming under the influence of cis-DDP]. / Charakterystyka wiazan DNA-bialko powstajacych pod wplywem cis-DDP.

In this article we described different nuclear proteins which have participated in DNA-protein cross-links induced by cis-DDP. The role of DNA-protein cross-links in the anticancer activity of cis-DDP is discussed.

Recent advances in FRET: distance determination in protein-DNA complexes.

Fluorescence resonance energy transfer (FRET) provides information on the distance between a donor and an acceptor dye in the range 10 to 100 A. Knowledge of the exact positions of some dyes with respect to nucleic acids now enables us to translate these data into precise structural information using molecular modeling. Advances in the preparation of dye-labeled nucleic acid molecules and in new techniques, such as the measurement of FRET in polyacrylamide gels or in vivo, will lead to an increasingly important role of FRET in structural and molecular biology.

Floppy SOX: mutual induced fit in hmg (high-mobility group) box-DNA recognition.

The high-mobility group (HMG) box defines a DNA-bending motif of broad interest in relation to human development and disease. Major and minor wings of an L-shaped structure provide a template for DNA bending. As in the TATA-binding protein and a diverse family of factors, insertion of one or more side chains between base pairs induces a DNA kink. The HMG box binds in the DNA minor groove and may be specific for DNA sequence or distorted DNA architecture. Whereas the angular structures of non-sequence-specific domains are well ordered, free SRY and related autosomal SOX domains are in part disordered. Observations suggesting that the minor wing lacks a fixed tertiary structure motivate the hypothesis that DNA bending and stabilization of protein structure define a coupled process. We further propose that mutual induced fit in SOX-DNA recognition underlies the sequence dependence of DNA bending and enables the induction of promoter-specific architectures.

SRYand architectural gene regulation: the kinetic stability of a bent protein-DNA complex can regulate its transcriptional potency.

Protein-directed DNA bending is proposed to regulate assembly of higher-order DNA-multiprotein complexes (enhanceosomes and repressosomes). Because transcriptional initiation is a nonequilibrium process, gene expression may be modulated by the lifetime of such complexes. The human testis-determining factor SRY contains a specific DNA-bending motif, the high-mobility group (HMG) box, and is thus proposed to function as an architectural factor. Here, we test the hypothesis that the kinetic stability of a bent HMG box-DNA complex can in itself modulate transcriptional potency. Our studies employ a cotransfection assay in a mammalian gonadal cell line as a model for SRY-dependent transcriptional activation. Whereas sex-reversal mutations impair SRY-dependent gene expression, an activating substitution is identified that enhances SRY's potency by 4-fold. The substitution (I13F in the HMG box; fortuitously occurring in chimpanzees) affects the motif's cantilever side chain, which inserts between base pairs to disrupt base pairing. An aromatic F13 cantilever prolongs the lifetime of the DNA complex to an extent similar to its enhanced function. By contrast, equilibrium properties (specific DNA affinity, specificity, and bending; thermodynamic stability and cellular expression) are essentially unchanged. This correlation between potency and lifetime suggests a mechanism of kinetic control. We propose that a locked DNA bend enables multiple additional rounds of transcriptional initiation per promoter. This model predicts the occurrence of a novel class of clinical variants: bent but unlocked HMG box-DNA complexes with native affinity and decreased lifetime. Aromatic DNA-intercalating agents exhibit analogous kinetic control of transcriptional elongation whereby chemotherapeutic potencies correlate with drug-DNA dissociation rates.

High progesterone levels and ciliary dysfunction--a possible cause of ectopic pregnancy.

OBJECTIVES: To investigate the effects of different levels of hormones on the ciliary activity of human oviducts and, consequently, to assess their possible role in tubal implantation of the fertilized egg. DESIGN: Fallopian tube epithelial samples were incubated in media with the addition of Estradiol (E2), progesterone (P), human menopausal gonadotropin (hMG), LH, or pure FSH (Metrodin) in different concentrations. The ciliary beat frequency (CBF) was measured after 24 h of incubation. Then the media were exchanged to media without the addition of hormones and the CBF was measured again 24 h later by using the photoelectric technique. SETTING: University teaching hospital, IVF unit. RESULTS: Twenty-four hr after the addition of P to the culture medium in concentrations of 0.5 or 1 ng/ml a significant decline of the CBF down to 63% of the control level was observed (P < 0.001) and with P in concentration of 2 ng/ml or greater, 50-70% of the cilia were paralyzed. These effects of P were found to be reversible. Incubation with E2 induced a slight increase of 4% in the mean CBF (P = 0.002). Twenty-four hr incubation with Metrodin, Pergonal, or LH did not affect ciliary motility. CONCLUSIONS: Higher levels of progesterone cause ciliary dysfunction and subsequently may be a possible cause of ectopic pregnancy.

Medical aspects of ketone body metabolism.

Ketone bodies are produced in the liver, mainly from the oxidation of fatty acids, and are exported to peripheral tissues for use as an energy source. They are particularly important for the brain, which has no other substantial non-glucose-derived energy source. The 2 main ketone bodies are 3-hydroxybutyrate (3HB) and acetoacetate (AcAc). Biochemically, abnormalities of ketone body metabolism can present in 3 fashions: ketosis, hypoketotic hypoglycemia, and abnormalities of the 3HB/AcAc ratio. Normally, the presence of ketosis implies 2 things: that lipid energy metabolism has been activated and that the entire pathway of lipid degradation is intact. In rare patients, ketosis reflects an inability to utilize ketone bodies. Ketosis is normal during fasting, after prolonged exercise, and when a high-fat diet is consumed. During the neonatal period, infancy and pregnancy, times at which lipid energy metabolism is particularly active, ketosis develops readily. Pathologic causes of ketosis include diabetes, ketotic hypoglycemia of childhood, corticosteroid or growth hormone deficiency, intoxication with alcohol or salicylates, and several inborn errors of metabolism. The absence of ketosis in a patient with hypoglycemia is abnormal and suggests the diagnosis of either hyperinsulinism or an inborn error of fat energy metabolism. An abnormal elevation of the 3HB/AcAc ratio usually implies a non-oxidized state of the hepatocyte mitochondrial matrix resulting from hypoxia-ischemia or other causes. We summarize the differential diagnosis of abnormalities of ketone body metabolism, as well as pertinent recent advances in research.

Progesterone response to exogenous gonadotropins after suppression with leuprolide acetate in gamete intrafallopian transfer.

Fifty-three cycle of ovarian stimulation with human menopausal gonadotropins after suppression of endogenous gonadotropin release with leuprolide acetate were completed for the purpose of performing gamete intrafallopian transfer. Thirty-seven biochemical pregnancies resulted, with 29 progressing to clinical status, for rates of 69.8% and 54.7%, respectively, per cycle. In comparing characteristics of the follicular recruitment phases of failed cycles to those of cycles resulting in pregnancies, no differences were observed in serum estradiol concentrations or follicular development. However, pregnancy rates were highly associated with peak concentrations of progesterone prior to the administration of human chorionic gonadotropin. Three levels of progesterone range were found on the basis of outcome: 6 high-progesterone cycles produced no pregnancies (0%), 32 mid-range progesterone cycles produced 31 pregnancies (97%), and 15 low-progesterone cycles produced six pregnancies (40%). The pregnancy rate in the last group improved when the luteal support was doubled.

Suppression of gonadotrophin secretion does not reverse premature ovarian failure.

The effect of prolonged inhibition of gonadotrophin secretion was studied in 12 women with premature ovarian failure. All the patients had plasma concentrations of follicle-stimulating hormone (FSH) greater than 20 i.u./l, and in six, primordial follicles had been seen on ovarian biopsy. Goserelin (Zoladex, ICI), a depot synthetic analogue of luteinizing hormone-releasing hormone (LHRH) was administered by three consecutive 4-weekly injections. Plasma concentrations of luteinizing hormone (LH) fell from 34 (SD 11) i.u./l to 2.4 (SD 1.9) i.u./l, and plasma concentrations of FSH fell from 106 (SD 29) i.u./l to 4.5 (SD 2.6) i.u./l 4 weeks after the first injection. Plasma concentrations of gonadotrophins returned to pretreatment values in every patient within 9 weeks of the final injection of goserelin. Regular ultrasonography during the period following the final injection failed to demonstrate the development of ovarian follicles in any patient, and plasma concentrations of oestradiol remained below 100 pmol/l. This study has failed to show that suppression of gonadotrophin secretion reverses premature ovarian failure.

Variation in the biologic and biochemical characteristics of human menopausal gonadotropin.

Lot differences in the biopotency of human menopausal gonadotropin (hMG) were evaluated and the potential biochemical basis was investigated. The in vivo biopotency of hMG was assessed by a unique bioassay that evaluates the number of ova shed in the cyclic hamster in response to hMG administration. Significant variation in hMG lots was observed using this assay. When subjected to chromatofocusing, hMG displayed five immunoreactive follicle-stimulating hormone (FSH) isohormones and nine luteinizing hormone (LH) isohormones. The relative distribution of FSH, but not LH isohormones, was slightly but significantly different between the lots tested. These data indicate that significant differences exist in the ability of commercially available hMG to stimulate follicular development and ovulation. The biochemical basis for these differences in in vivo biopotency remains to be elucidated.

Evidence for diverse structural variations of the forms of human FSH within and between pituitaries.

The biological activity of FSH in vitro (B-vitro) was compared with its radioimmunological (RIA) activity in 38 pituitary extracts from men and women. The B-vitro method was based upon the estimation of oestradiol produced by cultured Sertoli cells from 10-day-old rats. The mean B-vitro/RIA ratios for FSH of men and young and elderly women were almost identical. The mean values of median charge of the forms of FSH differed significantly between the three groups of human adults. The B-vitro/RIA ratio of different forms of FSH, separated by electrophoresis of 14 individual pituitary extracts, was higher for less negatively charged than for more negatively charged forms. Some forms of FSH with the same charge and separated from different pituitary extracts, e.g. from a young and an elderly woman, had significantly different B-vitro/RIA ratios. This ratio, thus, was not related to the charge per se of the hormone. However, the relation between the B-vitro/RIA ratio and the charge of FSH was similar for individual pituitary extracts when charge was expressed in relation to median charge. The results suggest that different molecular structural variations of FSH are involved in the polymorphisms observed within and between individual pituitaries. In human adults, the variation of FSH between the pituitaries mainly affects the metabolic clearance rate (MCR) of the hormone, whereas the variation within the individual pituitary affects both the MCR of the hormone and its biological effect at the target cell.

Deficiency of 3-methylglutaconyl-coenzyme A hydratase in two siblings with 3-methylglutaconic aciduria.

We studied two patients with 3-methylglutaconic aciduria in order to determine the molecular defect. A new assay for 3-methylglutaconyl-coenzyme A (CoA) hydratase has been developed in which the substrate, [5-14C]3-methylglutaconyl-CoA, was synthesized using 3-methylcrotonyl-CoA carboxylase purified from bovine kidney. In this assay the products of the reaction are isolated by reverse-phase high performance liquid chromatography and the rates of conversion from substrate are measured. The Michaelis constant for 3-methylglutaconyl-CoA in normal fibroblasts was 6.9 mumol/liter. The mean activity of 3-methylglutaconyl-CoA hydratase in control fibroblasts was 495 pmol/min per mg protein. In the two patients the values were 11 and 17 pmol/min per mg protein, or 2-3% of normal.