Mouse model for endometriosis is characterized by proliferation and inflammation but not epithelial-to-mesenchymal transition and fibrosis

Endometriosis is a common disorder of unknown etiology, and non-surgical therapies are still a challenge. Tounderstand the pathogenesis and preclinical testing of drugs for endometriosis, animal models are highlydesirous. Herein, we carried out longitudinal characterization of a mouse model for endometriosis whereuterine tissue was transplanted onto the intestinal mesentery. During the course of lesion development from day15 to 60 post-induction, the ectopic endometrium became pale, fluid-filled and the animals developed peritoneal adhesions. Most lesions resembled a well-differentiated type of endometriosis and * 13% of animalshad mixed type of lesions. There was extensive stromal compaction in the ectopic tissue. During the progression of endometriosis, there was increased proliferation of epithelial and stromal cells as evident by PCNAstaining. Cyp19a1 (aromatase) mRNA was detected in the ectopic lesions on day 15 and 30 post-induction ofendometriosis, by day 60 the expression was reduced. As compared to the control endometrium, the mRNAlevels of Esr1 progressively reduced while the levels of inflammation associated genes (Esr2, Ifng, Tnf andIl1b) increased in the ectopic lesions. Infiltration of macrophages and polymorphonuclear leucocytes was alsoobserved in the ectopic lesions indicative of inflammation. As compared to control, there was no change inlevels of Cytokeratin and E-cadherin in the epithelial cells of ectopic endometrium. We did not observeexcessive collagen deposition or a-SMA positive myofibroblasts in the stroma of the ectopic endometrium.Thus, epithelial-to-mesenchymal transition and fibrosis are not detected in the mouse model of endometriosis.Our results show that the mouse model of endometriosis mimics some but not all the features of humanendometriosis.

Study on the Marker Steroids of New Zealand Deer (Cervus elaphus var. scoticus) Velvet Antler by UPLC-MS/MS and HPLC-PDA Methods

Eleven steroid hormones (SHs: androstene-3,17-dione, estrone, β-estradiol, α-estradiol, testosterone, dehydroepiandrosterone, 17á-hydroxyprogesterone, medroxyprogesterone, megestrol acetate, progesterone, and androsterone) were detected from New Zealand deer (Cervus elaphus var. scoticus) velvet antler (NZA, 鹿茸). A method for the quantification of eleven SHs was established by using ultraperformance liquid chromatography (UPLC)-MS/MS. The linearities (R² > 0.991), limits of quantification (LOQ values, 0.3 ng/mL to 23.1 ng/mL), intraday and interday precisions (relative standard deviation: RSD 0.999), LOQ values (30 ng/mL to 350 ng/mL), intraday and interday precisions (RSD < 1.93%), and recovery rates (97.2% to 103.5%) for the three 7-O-CSs were determined. These quantitative methods are accurate, precise, and reproducible. As a result, it is suggested that the five steroid compounds of androstene-3,17-dione, androsterone, 7-ketocholesterol, 7α-hydroxycholesterol, and 7β-hydroxycholesterol could be marker steroids of NZA. These methods can be applied to quantify or standardize the marker steroids present in NZA.

Effects of Pim-1 inhibitor on mouse model of inflammatory bowel disease induced by TNBS / 中南大学学报(医学版)

Objective:To explore the role of Pim-1 in the pathology of inflammatory bowel disease and the potential effect of Pim-1 inhibitor on treating such disease.Methods:Forty-five BALB/c mice were randomly divided into 5 groups (n=9):A normal control group,a inflammatory bowel disease group,two different dose of Pim-1 inhibitor treatment groups,and steroidhormone treatment group.The model of inflammatory bowel disease was induced by intracolonic administration of 2,4,6-trinitrobenzenestdfonic acid (TNBS) and ethanol mixture.Mice were treated with Pim-1 inhibitor [intraperitoneal inject,5 or 10 mg/(kg.d)] for 5 days and prednisone (intragastric administration,0.1 mg/d) for 5 days.The DAI,colon length,gross score and pathological grade were evaluated.The expressions ofT cell master transcription factors T-box expressed in T cells (T-bet),GATA binding protein 3 (GATA-3),RA orphan receptorγ (RORyt)and forkhead box P3 (Foxp3) were measured by Real-time PCR and Western blot,respectively.Results:Pim-1 inhibitor and prednisone showed therapeutic effect on acute TNBS colitis in vivo.GATA3 and RORγt were significantly up-regulated in acute TNBS colitis (P＜0.05).In contrast,the expression of Foxp3 was suppressed in the inflammatory bowel disease group,whereas it did not cause any significant change in T-bet expression (P＞0.05).Administration of Pim-1 inhibitor and prednisone resulted in suppression of GATA3,RORγt expression,and the increase of Foxp3 expression (P＜0.05).Administration of Pim-1 inhibitor and prednisone resulted in inhibition of T-bet mRNA expression (P＜0.05),but only prednisone could inhibit T-bet protein expression (P＞0.05).Conclusion:Pim-1 inhibitor significantly suppresses Th2-and Th17-type immune responses.Furthermore,Pim-1 inhibitor could induce T-cell differentiation towards a Treg phenotype.Pim-1 inhibitor has therapeutic effect on acute TNBS colitis.

Research progress on analytical methods for blood steroid hormones / 中草药

Steroid hormone, the component of the endocrine system, plays an extremely important role in body development, immune regulation, and birth control. The accurate determination of steroid hormone is important for the prevention, diagnosis, and treatment of clinical endocrine diseases. In recent years, the determination of endogenous steroid hormones in biological samples has become increasingly widespread in clinical applications. In this article, the commonly used analysis methods of typical blood steroids from 2013 to 2018 were generalized and compared in order to provide more reference for the accurate determination of steroid hormone levels, and also provide more accurate basis for later clinical diagnosis and treatment.

The Hsp90 Chaperone Machinery: An Important Hub in Protein Interaction Networks.

Heat shock protein 90 (Hsp90) represents one of the most conserved proteins in living organisms and is present in all kingdoms of life except for Archaea. HSP90 proteins define a widespread family of molecular chaperones that play a fundamental role in protein homoeostasis and viability. HSP90s mediate folding and maturation of a broad spectrum of client proteins including steroid hormone receptors, transcription factors, and protein kinases. HSP90s primarily exist as homodimers whose activity is regulated by ATP. Hsp90 can adopt different ATP-triggered conformations, ranging from an open V-shaped unliganded to a closed ATP-bound state. HSP90 chaperones can be found not only in the cytosol, ER, chloroplasts, mitochondria, and the nucleus but also in the extracellular milieu where they act as potent stimulators of immune responses. The activity of Hsp90 is regulated by post-translational modifications and its association with numerous co-chaperones and client proteins involved in signal transduction and transcriptional regulation. Elevated levels of HSP90s can be found in a broad spectrum of cancers where they enhance cell growth, suppress senescence, and confer resistance to stress-induced apoptosis, including protection against cytostatic drugs and radiation therapy. Since numerous oncoproteins are clients of Hsp90, targeting Hsp90 represents a useful anti-cancer approach. In this review, the current knowledge on the Hsp90 chaperone machinery and its role in disease and therapy is compiled.

Steroid hormone receptors and prostate cancer: Role of structural dynamics in therapeutic targeting / 亚洲男科学杂志(英文版)

Steroid hormone receptors (SHRs) act in cell type- and gene-specific manner through interactions with coregulatory proteins to regulate numerous physiological and pathological processes at the level of gene regulation. Binding of steroid receptor modulator (SRM) ligand leads to allosteric changes in SHR to exert positive or negative effects on the expression of target genes. Due, in part, to the fact that current SRMs generally target ligand binding domain (LBD)/AF2 and neglect intrinsically disordered (ID) N-terminal domain (NTD)/AF1, clinically relevant SRMs lack selectivity and are also prone to the development of resistance over time. Therefore, to maximize the efficacy of SHR-based therapeutics, the possibility of developing unique modulators that act to control AF1 activity must be considered. Recent studies targeting androgen receptor′s (AR′s) ID AF1 domain for the castration-resistant prostate cancer has provided the possibility of therapeutically targeting ID NTD/AF1 surfaces by allosteric modulations to achieve desired effects. In this review article, we discuss how inter- and intra- molecular allosteric regulations controlled by AR′s structural flexibility and dynamics particularly the ID NTD/AF1 is an emerging area of investigation, which could be exploited for drug development and therapeutic targeting of prostate cancer.

Lc-ms/ms for determination of seven steroid hormones in sera of primary hepatocellular carcinoma patients / 第二军医大学学报

Objective To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of seven steroid hormones in sera of patients with primary hepatocellular carcinoma. Methods The serum samples were extracted with ethyl acetate. and then derivatized with dansyl chloride. Chromatographic column was Agilent Poroshell 120 EC-Ci8 (2. 1 mmX150 mm, 2. 7 μm) column, mobile phase was acetonitrile and 0. 1% formic acid-water solution, flow rate was 0. 3 mL/min. the column temperature was 35°C. and the MSdetection was selected in dynamic MRM mode. Results Seven steroid hormones (estradiol. estriol. estrone. hydrocortisone. testosterone. androstenedione and progesterone) were baseline separated within 8 min. which possessed good linear relationship (r>0. 99) within the linear concentration range; the intra-day and inter-day precisions were less than 15%. the recoveries were ranged from80% to 119%. the matrix effect was 85%-112%. The detection results were stable when the samples were examined 6 h after collection. 24 h after treatment at room temperature. after three freezethaw cycles. and 30 dayswhen stored at —80°C. Conclusion The current LC-MS/MS method is simple. accurate. sensitive and reproducible. and it canbe applied to determine steroidhormones in the sera of patients with primary hepatocellular carcinoma.

Influence of steroid hormone on expression of Bin1b in primary cultured epididymal epithelial cells / 第二军医大学学报

Objective To study the expression of Bin1b in primary cultured epididymal epithelial cells treated with different steroid hormones at various concentrations. Methods Expression of Bin1b mRNA was examined by agarose gel electrophoresis and PCR in primary cultured epididymal epithelial cells, which were treated with dihydrotestosterone (DHT, 10-9,10-8 and 10-7mol/L), estradiol (E2, 10-9 and 10-8mol/L)) or dexamethasone (Dex, 10-8 and 10-7mol/L) at different concentrations. Results The expression of Bin1b mRNA in primary cultured epididymal epithelial cells was decreased in a time-dependent manner. The mRNA level of Bin1b was decreased by 40% at day 2 and 70% at day 3, and it was hardly detectable at day 5. DHT up-regulated the expression of Bin1b mRNA in a dosage-dependent manner compared with corresponding vehicle control. 10-9mol/L DHT up-regulated Bin1b mRNA level by 37% compared with corresponding vehicle control(P<0.05), and it was further up-regulated after the treatment with 10-8mol/L and 10-7mol/L DHT. E2 and Dex at various concentrations showed no significant effects on the expression of Bin1b mRNA in primary cultured epididymal epithelial cells. Conclusion Dihydrotestosterone can up-regulate the expression of Bin1b mRNA in primary cultured epididymal epithelial cells, while estradio and dexamethasone can not.

Historical story on natural medicinal chemistry: Steroids / 中草药

Steroids are the most important class of natural products. In the history of the development of human beings, not only steroidal compounds have made the special contribution for human health, but also its special stereochemical structure plays an important role in the development of organic chemistry, especially organic chemistry theory, which could improve the stereochemical theory. In this paper, the discovery, biological activity, the stereochemical structure of important compounds, stereochemistry, and the synthesis of steroids are briefly introduced for the purpose to broaden their horizon and provide some ideas for young teachers in the teaching and scientific research.

Research progress in steroid hormone analysis in vivo by chromatography-mass spectrometry / 药学实践杂志

Steroid hormones were chemical compounds of high efficiency , which played a significant role in various physiologi-cal activities.However, the concentrations of hormones in vivo were extremely low, so it was very important for their accurate quantifi-cation.Chromatography-mass spectrometry technology was widely employed because of advances in high efficient , rapid and sensitive. The application of steroid hormone analysis by chromatography-mass spectrometry was reviewed in this paper , which could provide ref-erence for furthuer clinical research .

Important role of the C-terminal region of pig aldo-keto reductase family 1 member C1 in the NADPH-dependent reduction of steroid hormones.

The NADPH-dependent reduction activities of two paralogous pig AKR1C1s with and without 19 additional amino acid residues in C-terminus were evaluated against steroid hormones including 5a-dihydrotestosterone, testosterone, progesterone, androstenedione and 5a-androstane-3,17-dione, which act as substrates of the AKR1C1s. Among the hormones, the AKR1C1s exhibited the highest activity against 5a-dihydrotestosterone and the lowest activity against testosterone and progesterone. Furthermore, the AKR1C1s showed the largest differential activities against 5a-dihydrotestosterone, but no such change of activities was found against progestrone and testosterone. These results suggest that the C-terminal region of AKR1C1 plays an important effect in the reduction activities of pig AKR1C1. Thus, the differential activities of two AKR1C1 paralogs observed in the present study provide important insights in understanding the molecular evolution.

Progress of research in adrenal cortical cells model / 中华内分泌代谢杂志

This paper describes the biological characteristics of human and mouse adrenal cortical carcinoma cell lines and presents research progress made in rat and bovine primary cultured adrenal cortical cells.The effect of angiotensin Ⅱ,potassium,adrenocorticotropic hormone and cAMPon different adrenal cortical cell lines were compared and analyzed,regarding the differences of steroid hormone secreted.The article suggests that it is necessary to develop and establish some ideal adrenal cortex cell models for furtherstudying the mechanisms of steroid hormone synthesis and secretion.

Hormonal regulation of uterine chemokines and immune cells / 대한생식의학회지

The ultimate function of the endometrium is to allow the implantation of a blastocyst and to support pregnancy. Cycles of tissue remodeling ensure that the endometrium is in a receptive state during the putative 'implantation window', the few days of each menstrual cycle when an appropriately developed blastocyst may be available to implant in the uterus. A successful pregnancy requires strict temporal regulation of maternal immune function to accommodate a semi-allogeneic embryo. To preparing immunological tolerance at the onset of implantation, tight temporal regulations are required between the immune and endocrine networks. This review will discuss about the action of steroid hormones on the human endometrium and particularly their role in regulating the inflammatory processes associated with endometrial receptivity.

Changes of sex hormone levels and sexual function in male patients after liver transplantation / 中华普通外科杂志

ObjectiveTo investigate changes of sex hormone levels and sexual function in male patients after liver transplantation.MethodsBlood samples were taken from 69 male patients receiving liver transplantation pretransplantation and postoperative 3rd,7th,14th day,1st and 3rd month respectively.Serum estradiol and testosterone were measured by radioimmunoassay.Serum sex hormone binding globulin was measured by ELISA.Sixty-nine discharged adult male liver transplant recipients ( age ranging 24 -45 years) with a normally functioning allograft for at least 6 months were questioned on their pre-and post-operativesexualfunction compared with 23adulthealthy controls.ResultsPretransplantation serum estradiol and sex hormone binding globulin were significantly higher (87.56±31.21 vs.26.00 ±9.12,u=9.30,P＜0.0001,134.50±30.68 vs.51.04 ±12.05,u=12.69,P ＜ 0.0001,respectively),and testosterone was lower than normal controls (2.02 ± 1.28 vs.4.82 ± 1.48,u =-8.73,P ＜0.0001 ) and levels were correlated with MELD scores (r =0.80,P ＜0.0001,r =-0.77,P ＜0.0001,r =0.72,P ＜0.0001,respectively).Sex hormone levels were back to normal on postoperative 2nd week (serum estradiol) or 1st month (serum testosterone and sex hormone binding globulin).Compared with pretransplant status,sexual function improved significantly when evaluated on postoperative 6th month.ConclusionsLevels of serum sex hormone were correlated with MELD scores before liver transplant.After liver transplant,estradiol was back to normal level on postoperative 2nd week,testosterone and sex hormone binding globulin back to normal level on postoperative 1 st month.Sexual function significantly improves after liver transplantation.

Effects of combined administration of low dose gossypol with steroid hormones on spermatocyte apoptosis and phagocytosis of sertoli cells / 解剖学报

ObjectiveTo investigate effects of combined regimen of low dose gossypol with steroid hormones on apoptosis of spermatocyte and phagocytosis of sertoli cells. Methods Adult male rats were divided into four groups randomly, group GH: rats were fed orally with gossypol(GA) [ 12.5mg / (kg·d) ] and desogestrel(DSG)[125μg/(kg·d) ]/ethinyloestradil(E)[25μg/ (kg·d) ]/testosterone undecanoate(TU)[100mg/(kg·d) ]; group G: a single does of GA[12.5mg/(kg·d ) ] was given; group H: DSG[125μg/(kg·d) ]/E[25μg/(kg·d) ]/TU[100mg/(kg·d) ] were administered; group C: rats only received vehicle(1% methyl cellulose). Testes from all the rats were removed at 4, 6 and 8 weeks after treatment for counting of spermatocyte and round spermatid using stereological assay, and for detecting apoptosis of spermatocyte and phagocytosis of sertoli cell by TUNEL and oil red O staining respectively. Results In GH group, the number of spermatocyte and round spermatid were reduced, while apoptosis of spermatocyte and staining of oil red O in seminiferous epithelium increased significantly. All changes were caused by steroid hormones in the combined regimen. Conclusion Induction of apoptosis of spermatocyte and then being phagocytosed by Sertoli cell is one of antifertility mechanisms of the combined regimen.

The Effect of Collagen Supplementation from Pork Skin on Serum Collagen, Serum Sex Steroid Hormone, Serum Lipid and Skin Crack in Korean Middle-aged Women / 대한지역사회영양학회지

This study was performed to examine if the effects of collagen supplementation from pork skin could improve the sex steroid hormone, serum lipid and skin crack in Korean middle-aged women. Middle-aged women (40-55 years) who were not diagnosed with any type of disease were included in this study and thirty subjects were randomly assigned to a control group (n = 15) or a collagen supplemented group (n = 15). The collagen supplemented group ingested collagen flour 2 g, 3 times a day for 12 weeks. We measured serum collagen, estrogen, estradiol, estriol, progesterone, total cholesterol, triglyceride, HDL-cholesterol and LDL-cholesterol concentration. The collagen supplementation group had significantly increased serum collagen (p < 0.05) compared with the control group. In addition, skin crack was improved. But, there were no differences for sex steroid hormone and lipid profile in control and collagen supplemented groups. The result of the present study demonstrated that supplementation of 6 g collagen per day for 12 weeks can give beneficial effects on skin crack reduction and serum collagen concentration.

Alteration of Estrogen Receptor, Progesterone Receptor, and HER-2 Expression in Breast Cancer after Neoadjuvant Chemotherapy / 한국유방암학회지

PURPOSE: We aimed to assess the concordance of the immunohistochemical profiles of core biopsy before administrating neoadjuvant chemotherapy with that of the surgical specimens after a definitive operation for breast cancer. METHODS: We retrospectively reviewed the estrogen receptor (ER), progesterone receptor (PR), and HER-2 expressions in 130 consecutive patients who received neoadjuvant chemotherapy and were followed by surgery during the period between February 2002 and March 2006. The pathologic complete tumor response rate for this group was 4.6% (6/130). Both the pre- and post-operative immunohistochemical profiles were available in 32 of the 124 patients (25.8%). Immunohistochemical staining was done on the core biopsies before chemotherapy and on the surgical specimens after operation. RESULTS: There were 12 markers from 11 patients that were altered out of the 96 total markers (ER, PR, or HER-2) from 32 patients: 2 ER (2/12, 16.7%), 4 PR (4/12, 33.3%), and 6 HER-2 (6/12, 50.0%). One patient simultaneously had changes in the expressions of PR and HER-2. Conversion of the hormone receptor status occurred in 3 patients (3/32, 9.4%): this was positive to negative in two, and vice versa in one. In addition, there were 6 conversions (6/32, 18.8%) of the HER-2 status from negative to positive. CONCLUSION: The hormone receptor status changed in 9.4% of the 32 patients and the HER-2 status changed in 18.8% of the 32 patients after neoadjuvant chemotherapy. We have concluded that conducting only a single immunohistochemical study about ER, PR, and HER-2 may not be enough to exactly estimate the tumor marker status in the neoadjuvant setting.

Cerebral Aneurysm and Aortic Coarctation in a 46, XY Female. Is it Causal or Coincidental?

Most vascular disorders tend to affect both the brain and heart, and among them, a clinical syndrome constituting cerebral aneurysm and aortic coarctation(AC) has been well recognized. Persistent hypertensive impact to the cerebral vasculature with developmental anomaly of the neural crest, precursor of ectomenchymal, would be closely associated with development of the cerebral aneurysm in AC. Gonadal steroid hormone, a guardian of the cardiovascular system, has been known for its protective effects on the vascular wall. Gonadal steroid hormone (androgen) insensitivity such as 46, XY female syndrome may increase the risk of hypertention and subsequent vascular anomalies. The authors report on a 46-year-old 46, XY female patient with AC who underwent surgical clipping of the ruptured cerebral aneurysm. Clinical implications and proposed pathogenetic mechanisms of aneurysm in this intersex syndrome are presented and discussed.

Steroid Receptor Coactivator Family and Breast Cancer / 中国普外基础与临床杂志

Objective To investigate the effect of steroid receptor coactivator family in initiation, development, treatment and prognosis of breast cancer. Methods The literatures in recent years which have related to the effect of steroid receptor coactivators in breast cancer are reviewed. Results Steroid receptor coactivators are essential for several kinds of steroid hormones binding to steroid receptors, so they are important accessory factors that induce the initiation, development and recurrence of breast cancer, and predictive factors that estimate the prognosis of breast cancer. Conclusion Inhibition of the expression and signaling pathway of steroid receptor coactivators may be effective for breast cancer prevention and treatment.

Testes biopsy analysis and hormonal reproduction levels of pig tail macaque (Macaca nemestrina) injected with testosterone enanthate (TE) and depot medroxy progesterone acetate (DMPA).

The objective of this study is to analyse the testes biopsy and to know the lnrmone level reproduction of pig tail macaque (Macaca nemestrina) injected with testosterone enanthate (TE) and depot medroxyprogesterone acetate (DMPA). Six pig tail male macaques, age 6 - 8 years old were used as sanrple. Three months after adaptation period, each animal was injected intra muscularly with 32 mg TE each week starting at week zero up to the sixth week. The treatment was continued every 3 weeks after the sixth week up to the 24th week. 40 mg of DMPA was injected intramuscularly at week zero, and continued every week up to week 18. Volume of the testes was taken every three weeks and blood samples for examination of gonadotropin hormone and steroid hormone were taken at 6 week intervals. Testes biopsy was perfomed at week 30 and week 48. Preparation of testes histological slides were made using the paraffin method and stained with hematoxylin-Eosin (HE). The results of this study showed that both testes volume decreased i.e. 18.35 cm3 ± 9.35 and 19.02cm3 ± 10.88 (at week zero) to 6.70 cm3 ± 3.80 and 7.02 cm3 ± 4.61 (lowest volume at week 21). In recovery period, the testes volume increased to 20.34 cmr ± 7.87 and 21.75 cm3 ± 7.09. The diameter of the seminiferous tubules and the score of spermatogenesis (right and left testes) at week 30 were 0.13 mm ± 0.027 and 0.13 mm ± 0.026 and score were 5.08 ± 2.67 and 5.41 ± 2.51. At week 48, both diameter of seminiferous tubules and spermatogenesis score increased to become 0.18 mm ± 0.029 and 0.18 mm ± 0.026, and score were 7.51 ± 2.14 and 7.57 ± 1.59, During this period, hyalinization and fibrosis of seminiferous tubule occurred. By week 6, the total testosterone, free testosterone, and estrogen hormone levels increased quite sharply and then decreased but still higher than base levels of hormone. In the recovery period, estrogen hormone increased significantly until the end of observation (week 48). FSH and LH hormone levels decreased until week 6, then the FSH hormone levels increased until the end of observation, while the LH hormone level is still lower than base level. Conclusion of this study is the injection of TE and DMPA combination will alter the histological structure of the pig tailed macaque testes i.e. decreasing the diameter of the seminiferous tubules, suppressing spermatogenesis and hyalinisation and fibrosis of seminiferous tubules. The damages of this structure are likely caused by inhibition of feedback mechanism of hypothalamus-hypophysis-testes.