Identification and validation of anoikis-related lncRNAs for prognostic significance and immune microenvironment characterization in ovarian cancer.

Anoikis, a form of apoptotic cell death resulting from inadequate cell-matrix interactions, has been implicated in tumor progression by regulating tumor angiogenesis and metastasis. However, the potential roles of anoikis-related long non-coding RNAs (arlncRNAs) in the tumor microenvironment are not well understood. In this study, five candidate lncRNAs were screened through least absolute shrinkage and selection operator (LASSO), and multivariate Cox regression analysis based on differentially expressed lncRNAs associated with anoikis-related genes (ARGs) from TCGA and GSE40595 datasets. The prognostic accuracy of the risk model was evaluated using Kaplan-Meier survival analysis and receiver operating characteristic (ROC) curves. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene set enrichment analysis (GSEA) analyses revealed significant differences in immune-related hallmarks and signal transduction pathways between the high-risk and low-risk groups. Additionally, immune infiltrate analysis showed significant differences in the distribution of macrophages M2, follicular T helper cells, plasma cells, and neutrophils between the two risk groups. Lastly, silencing the expression of PRR34_AS1 and SPAG5_AS1 significantly increased anoikis-induced cell death in ovarian cancer cells. In conclusion, our study constructed a risk model that can predict clinicopathological features, tumor microenvironment characteristics, and prognosis of ovarian cancer patients. The immune-related pathways identified in this study may offer new treatment strategies for ovarian cancer.

Immune system changes in those with hypertension when infected with SARS-CoV-2.

The coronavirus disease 2019 (COVID-19) outbreak has become an evolving global health crisis. With an increasing incidence of primary hypertension, there is greater awareness of the relationship between primary hypertension and the immune system [including CD4+, CD8+ T cells, interleukin-17 (IL-17)/T regulatory cells (Treg) balance, macrophages, natural killer (NK) cells, neutrophils, B cells, and cytokines]. Hypertension is associated with an increased risk of various infections, post-infection complications, and increased mortality from severe infections. Despite ongoing reports on the epidemiological and clinical features of COVID-19, no articles have systematically addressed the role of primary hypertension in COVID-19 or how COVID-19 affects hypertension or specific treatment in these high-risk groups. Here, we synthesize recent advances in understanding the relationship between primary hypertension and COVID-19 and its underlying mechanisms and provide specific treatment guidelines for these high-risk groups.

A deep learning-based system for assessment of serum quality using sample images.

BACKGROUND: Serum quality is an important factor in the pre-analytical phase of laboratory analysis. Visual inspection of serum quality (including recognition of hemolysis, icterus, and lipemia) is widely used in clinical laboratories but is time-consuming, subjective, and prone to errors. METHODS: Deep learning models were trained using a dataset of 16,427 centrifuged blood images with known serum indices values (including hemolytic index, icteric index, and lipemic index) and their performance was evaluated by five-fold cross-validation. Models were developed for recognizing qualified, unqualified and image-interfered samples, predicting serum indices values, and finally composed into a deep learning-based system for the automatic assessment of serum quality. RESULTS: The area under the receiver operating characteristic curve (AUC) of the developed model for recognizing qualified, unqualified and image-interfered samples was 0.987, 0.983, and 0.999 respectively. As for subclassification of hemolysis, icterus, and lipemia, the AUCs were 0.989, 0.996, and 0.993. For serum indices and total bilirubin predictions, the Pearson's correlation coefficients (PCCs) of the developed model were 0.840, 0.963, 0.854, and 0.953 respectively. Moreover, 30.8% of serum indices tests were deemed unnecessary due to the preliminary application of the deep learning-based system. CONCLUSIONS: The deep learning-based system is suitable for the assessment of serum quality and holds the potential to be used as an accurate, efficient, and rarely interfered solution in clinical laboratories.

Interactions Between Specific Immune Status of Pregnant Women and SARS-CoV-2 Infection.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen responsible for the Coronavirus Disease 2019 (COVID-19) global pandemic. Because it is a new and highly contagious coronavirus, most people, especially pregnant women, lack immunity. It is therefore important to understand the interaction between why pregnant women are susceptible to SARS-CoV-2 and the specific immune systems of pregnant women. Here, we provide an overview of the changes that occur in the immune system during pregnancy, the activation and response of the immune system in pregnant women with COVID-19, adverse pregnancy outcomes in pregnant women with COVID-19, and the treatment and prevention of COVID-19 in this population.

Analysis of the reproductive outcomes and the size of the unicornuate uterus measured by magnetic resonance imaging and their relationship.

PURPOSE: To evaluate the relationship between the uterine size measured by pelvic magnetic resonance and reproductive outcome in women with a unicornuate uterus. METHODS: This was a retrospective study including 140 patients affiliated with unicornuate uterus diagnosed by the pelvic MR prior to their first pregnancy in the Obstetrics and Gynecology Hospital of Fudan University from April 2010 to December 2017. All the length of the unicornuate uterus were re-measured and recorded by skilled radiologists during the study period. We divided all the 140 participants with complete pelvic MR imaging into four groups by the best reproductive outcomes, which refers to Group 1 (primary infertility, n = 21), Group 2 ( < 24 weeks' gestation, n = 34), Group 3 (preterm delivery, 24-35 weeks' gestation, n = 13), Group 4 ( ≥ 35 weeks' gestation, n = 72), followed them up and then analyzed the data. RESULTS: Measurements of 140 patients with hemi-uteri were retrieved for analysis. The mean length of the uterine was 4.90  ± 0.56 cm. There were no significant differences in the uterine cavity length, cervical length, endometrial thickness and uterine wall thickness between the four groups while the uterine length (P = 0.001) was statistically significant. Women with uterine lengths ≥  4.5 cm were more likely to experience full-term delivery compared with the other group (P = 0.001). Ordinal multiple logistic regression analysis showed that the uterine length [OR = 9.03 (95% CI: 2.90-28.13)] and uterine cavity length [OR = 0.32 (95% CI: 0.06-2.04)] were independent protective factors for better obstetric outcomes CONCLUSION: The uterine length is a reliable prognostic factor for the gestational week of delivery and an appropriate antenatal surveillance factor of women with unicornuate uterus.

A Multi-center, Randomized, Controlled and Open Clinical Trial of Heyan Kuntai Capsule () and Hormone Therapy in Perimenopausal Women.

OBJECTIVE: To evaluate the efficacy and safety of Heyan Kuntai Capsule (, HYKT) and hormone therapy (HT) on perimenopausal syndromes (PMSs). METHODS: From 2005 to 2008, 390 women with PMSs were recruited from 4 clinic centers. The inclusion criteria included ages 40 to 60 years, estradiol (E2) below 30 ng/L, and follicle stimulating hormone (FSH) above 40 IU/L, etc. The patients were randomly assigned to HYKT group or HT group by random number table method, administrated HYKT or conjugated estrogen with/without medroxyprogesterone acetate tablets for 12 months. During treatment, the patients were interviewed quarterly, Kupperman Menopausal Index (KMI) scores, hot flush scores, insomnia scores, Menopause-Specific Quality of Life (MENQOL) scores and adverse effects were used for evaluating drug efficacy and safety respectively. The last interview was made at the end of 12-month treatment RESULTS: After treatment, KMI scores of HYKT group and HT group were both significantly decreased compared with baseline (P <0.01) and there was no significant difference between groups (P >0.05), except that KMI of HYKT group was higher after 3-month treatment (P <0.05). After treatment, hot flush and insomnia scores were both improved significantly in two groups (P <0.01); and HT had a better performance than HYKT in improving hot flush (P <0.05). MENQOL were significantly improved in both groups after treatment (P <0.01); but there was no significant difference between two groups (P >0.05). The incidence of adverse event in the HYKT group was much lower than that in the HT group (P <0.01). CONCLUSIONS: HYKT could effectively relieve PMSs and improve patients quality of life without severe adverse reactions. Although HYKT exerted curative effects more slowly than hormone, it possessed better safety profile than hormone.

Initiation of GnRH agonist treatment on 3-5 days postoperatively in endometriosis patients: a randomized controlled trial.

Seventy patients with stage III or IV endometriosis were randomly assigned to 2 groups after conservative surgery. Group O (n = 35) received 3 cycles of a 28-day gonadotropin-releasing hormone agonist (GnRH-a) treatment (goserelin, 3.6 mg) starting 3-5 days postoperatively. Group M (n = 35) received the same treatment starting on days 1-5 of menstruation. Groups were further subdivided according to add-back treatment. Pre- and posttreated levels of estradiol (E2 ), follicle stimulating hormone (FSH), and luteinizing hormone (LH) and visual analog scale (VAS), Kupperman menopausal index (KMI), and bone mineral density (BMD) scores were recorded. The incidence of uterine bleeding was assessed. In both groups, serum levels of E2 , FSH, and LH and VAS scores decreased significantly after treatment. Spotting was the most frequent bleeding pattern. During cycle 1, the bleeding time in group M was much longer that than that in group O (P =.001), and the bleeding rate in group M was significantly higher than that in group O (P =.024, RR = 1.185). In patients with stage III or IV endometriosis, the efficacy of GnRH-a initiated 3-5 days postoperatively was equivalent to that of GnRH-a initiated on days 1-5 of menstruation. Female patients who initiated GnRH-a treatment 3-5 days postoperatively experienced less uterine bleeding during the first cycle of treatment.

Human Adenomyosis Endometrium Stromal Cells Secreting More Nerve Growth Factor: Impact and Effect.

Abnormal expression of nerve growth factor (NGF) was found in adenomyosis (AM). We collected AM foci from patients and eutopic endometrium from non-AM controls. Endometrium stromal cells (ESCs) were cultured. Different levels of 17ß-estradiol, tumor necrosis factor (TNF), CoCl2, and H2O2 were added to the culture system separately, then the expression level of NGF in ESCs was detected. After adding different levels of NGF, the proliferation and apoptosis of ESCs and aromatase expression were detected. We found that 17ß-estradiol promoted NGF production in AM ESCs but not in control ESCs; TNF promoted NGF production in both AM and control ESCs; and CoCl2 inhibited NGF production in control ESCs, but had no effect in AM ESCs. Nerve growth factor promoted the proliferation and synthesis of aromatase in AM ESCs. In conclusion, locally increased estrogen levels and inflammation may cause increased NGF production in the uterus of patients with AM. Nerve growth factor stimulated the proliferation and increased aromatase expression of ESCs from AM foci, suggesting NGF might contribute to the pathology and etiology of AM.

[Expression of nerve growth factor produced by ectopic endometrium from patients with adenomyosis and its relationship with pain scales and innervation].

OBJECTIVE: To investigate the expression of nerve growth factor (NGF) in the ectopic endometrium in adenomyosis patients, and explore the relationship between NGF expression and innervation or pain scales. METHODS: From Mar. 2009 to Oct. 2009, 45 adenomyosis patients undergoing hysterectomy in Obstetrics and Gynecology Hospital of Fudan University were enrolled in this study, which were classified into 33 cases in pain group and 12 cases in non-pain group based on symptom. The degree of dysmenoreal, chronic pelvic pain and dyspareunia was evaluated by visual analogue scale, including no pain, mild to moderate pain and severe pain group. In the mean time, 26 patients with leiomyoma or cervical intraepithelial neoplasia III (CIN III) undergoing hysterectomy were defined as control group. Ectopic endometrium from experimental group and eutopic endometrium from control group were collected in the surgery. The expression of NGF was examined by immunohistochemistry. The density of protein gene product (PGP) 9.5 positive nerve fibers was detected by immuno-fluorescence. RESULTS: The NGF level and the density of PGP 9.5 positive nerve fibers in adenomyosis pain group (0.25 ± 0.08, 16 ± 8 )were higher than adenomyosis painless (0.19 ± 0.05, P = 0.007;11 ± 5, P = 0.018) and control group (0.18 ± 0.05, P = 0.000; 9 ± 4, P = 0.000) . The NGF level and the density of PGP9.5 positive nerve fibers in severe dysmenorrheal group (0.29 ± 0.07, 19 ± 10) were higher than mild to moderate dysmenorrheal (0.22 ± 0.07, P = 0.018;13 ± 4, P = 0.035) and painless group (0.18 ± 0.05, P = 0.000;11 ± 5, P = 0.006) of adenomyosis patients. There was no difference of NGF level and the density of PGP 9.5 positive nerve fibers in chronic pelvic pain group and no chronic pelvic pain group of adenomyosis patients, so was dyspareunia group and no dyspareunia group. CONCLUSION: The increased NGF level of adenomyosis nodules and improving innervation might be involved in the mechanism of adenomyosis related pain.

The incidence and characteristics of uterine bleeding during postoperative GnRH agonist treatment combined with estrogen-progestogen add-back therapy in endometriosis patients of reproductive age.

To evaluate the incidence and characteristics of uterine bleeding during postoperative gonadotropin-releasing hormone agonist (GnRHa) treatment combined with the lowest effective dose of estrogen-progestogen add-back therapy in Chinese women of reproductive age with endometriosis. Seventy Chinese women aged 18 to 50 years with stage III or IV endometriosis and treated with postoperative GnRHa after conservative surgery for endometriosis were eligible for this study. Patients were randomly divided into two equal groups, G and A. Group G (n = 35) received three 28-day cycles of postoperative GnRHa treatment by subcutaneous injection (goserelin, 3.6 mg). Group A (n = 35) received the same GnRHa treatment in addition to daily estradiol valerate (0.5 mg) and dydrogesterone (5 mg) add-back therapy. Serum E2 and FSH levels were assessed at the end of each treatment cycle, as well as incidence and patterns of uterine bleeding. After the last GnRHa treatment cycle, endometrial thickness was evaluated by ultrasonography and the recovery of menstruation was recorded. Uterine bleeding incidence was above 90% in both groups during the first treatment cycle (group G: 90.6%; group A: 93.8%), but decreased markedly in the second treatment cycle (group G: 15.6%; group A: 21.9%), and continued to decline until the end of the third treatment cycle (group G: 6.3%; group A: 12.5%). For each cycle, the incidence of uterine bleeding in group A was slightly but not statistically higher. Irregular spotting was the most common uterine bleeding pattern observed in each of the three treatment cycle. The addition of estrogen and progestogen therapy to a postoperative GnRHa regimen does not lead to an increase in the duration or amount of treatment-induced uterine bleeding.

Benign and malignant ovarian steroid cell tumors, not otherwise specified: case studies, comparison, and review of the literature.

Ovarian steroid cell tumors, not otherwise specified (NOS) are rare sex cord-stromal tumors of the ovary with malignant potential. So far only a few cases were reported in English literature through the Pubmed search. Here we report two cases of such tumor, one was benign (first case underwent laparoscopic cystectomy) and the other was malignant (died 10 months later after initial diagnosis), both presented with amenorrhea and clinical signs or symptoms of virilization. In malignant case, we provided evidence (tumor embolus) in addition to the reported five characteristics associated with malignancy. On further evaluation, laboratory investigations revealed hyperandrogenism in the male range, while follicle stimulating hormone (FSH) and luteinising hormone (LH) levels were within normal limits. Various aspects of the presentation, diagnosis, and treatment of these tumors are discussed.

Marker gene screening for human mesenchymal stem cells in early osteogenic response to bone morphogenetic protein 6 with DNA microarray.

AIMS: Microarray data were analyzed using bioinformatic tools to screen marker genes of human mesenchymal stem cells (hMSC) in response to bone morphogenetic protein 6 (BMP6). RESULTS: A total of 190 differentially expressed genes were identified. The interaction network was divided into three functional modules. These genes were connected with BMP signaling pathways and regulation of cell processes, while NOG and BMPR2 participated in the transforming growth factor-beta signal pathway. Besides, several related small molecules were acquired. CONCLUSION: Marker genes in osteogenic responses to BMP6 treatment for hMSC were screened with microarray data along with elaborate function analysis by bioinformatics. NOG and BMPR2 showed potential to become indicators to monitor the directed differentiation of hMSC into osteoblasts, which can be used for bone disease treatment. Moreover, small molecules such as W-13 were retrieved and provided directions for future drug design.

The relationship between endogenous androgens and body fat distribution in early and late postmenopausal women.

OBJECTIVES: To investigate the relationship between endogenous androgens and body fat distribution in early and late postmenopausal women. MATERIALS AND METHODS: We enrolled postmenopausal women consisting of an early group (≤ 5 years since menopause, n = 105) and a late group (≥ 10 years since menopause, n = 107). Each group was subdivided into normal weight (BMI <24 kg/m(2)) group, overweight and obese (BMI ≥ 24 kg/m(2)) group. Fasting total testosterone (T), dehydroepiandrosterone-sulfate (DHEA-S) and sex hormone-binding globulin (SHBG) levels were measured. Body fat distribution was evaluated by dual-energy X-ray absorptiometry (DEXA). RESULTS: Late postmenopausal women had a higher proportion of body fat than early postmenopausal women. The body fat of the overweight and obese women had a greater tendency to accumulate in the abdomen compared with the normal weight women both in early and late postmenopausal groups. The overweight and obese women had a higher free testosterone (FT) than the normal weight women in early postmenopausal women (P<0.05). In late postmenopausal women, the overweight and obese women had higher DHEA-S levels than normal weight women (P<0.05). No direct relationship was observed between the T levels and body fat distribution both in early and late postmenopausal groups (P>0.05).The FT in early postmenopausal women and the DHEA-S levels in late postmenopausal women correlated positively with the trunk/leg fat ratio (T/L) and the proportion of android fat whereas correlated negatively with the proportion of gynoid fat in the partial correlation and multiple linear regression analyses (all P<0.05). CONCLUSIONS: Serum T levels do not correlate directly with body fat distribution, the FT in early postmenopausal women and DHEA-S levels in late postmenopausal women correlate positively with abdominal fat accumulation.

Oral continuous combined 0.5 mg estradiol valerate and 5 mg dydrogesterone as daily add-back therapy during post-operative GnRH agonist treatment for endometriosis in Chinese women.

OBJECTIVE: To evaluate the lowest effective dose of combined estrogen and progestogen (E(2)+P) add-back therapy during post-operative gonadotropin-releasing hormone agonist (GnRHa) treatment for endometriosis in Chinese women. STUDY DESIGN: The study enrolled 81 patients aged 18 to 50 years with stage III or IV endometriosis, as diagnosed by surgery. All patients were given GnRHa 3.6 mg by subcutaneous injection once every 28 days for a total of three times. Patients were divided into three groups: the first (n = 35; GnRHa only group) received GnRHa only without add-back therapy, the second (n = 35; 0.5 mg E(2)+P add-back group) received GnRHa plus 0.5 mg estradiol valerate and 5 mg dydrogesterone orally every day, and the third (n = 11; 1 mg E(2)+P add-back group) received GnRHa plus 1 mg estradiol valerate and 10 mg dydrogesterone orally every day for the duration of treatment. All patients were required to follow up at our hospital at 4, 8 and 12 weeks after treatment initiation to assess efficacy and levels of serum reproductive hormones. RESULTS: Compared with baseline levels, serum levels of the four reproductive hormones assessed (E(2), LH, P(4) and FSH) were significantly decreased in both the GnRHa only and the 0.5 mg E(2)+P add-back groups at 4, 8, and 12 weeks after treatment; and levels reached a stable state at 4 weeks of treatment. In the 1 mg E(2)+P add-back group, LH and FSH serum levels were significantly decreased, while those of E(2) and P were not significantly different at any of the time points assessed. In the 0.5 mg E(2)+P add-back group, E(2) serum levels decreased drastically at first, then gradually over the course of the study. In contrast, pre- and post-treatment E(2) serum levels in the 1 mg E(2)+P add-back group were not significantly different, and these levels were over 45 pg/mL for the entire study duration. Comparison among groups showed that E(2) levels in both add-back groups were significantly higher than in the GnRHa only group at 12 weeks after treatment. Furthermore, E(2) serum levels in the two add-back groups at 8 and 12 weeks after treatment were significantly different. CONCLUSION: Oral continuous combined 0.5 mg/d estradiol valerate and 5 mg/d dydrogesterone as immediate add-back therapy during post-operative GnRH agonist treatment for severe endometriosis may be the most suitable regimen for Chinese women.

[Survey on characteristics of menopause of Chinese women with the age of 40-60 years at gynecological clinic from 14 hospitals].

OBJECTIVE: To investigate the characteristics of menopause of Chinese women with the age of 40-60 years concerning gynecologic clinics in China. METHODS: From Mar.2008 to Sept.2008, a face-to-face questionnaire was conducted in gynecological clinic in perimenopausal and postmenopausal women in 14 hospitals in China, which included general demographic data, menstrual change process, climacteric symptoms and knowledge about menopause. Modified Kupperman index were used to evaluate climacteric symptoms during the recent week and awareness of hormonal replacement therapy were studied. RESULTS: A total of 1641 women were investigated. The ages of onset of menopause transition, climacteric symptoms and natural menopause were (47 ± 4), (46 ± 4), (49 ± 3) years old respectively. Climacteric symptoms could be found in 78.43% (1287/1641) women during menopausal transition, which were mainly mild to moderate symptoms. The top 5 symptoms were fatigue and weakness (71.48%, 1173/1641), irritability (68.68%, 1127/1641), insomnia (67.65%, 1110/1641), muscle and joint pain (64.11%, 1052/1641) and hot flush (57.90%, 950/1641). The climacteric symptoms were not constant during menopausal transition, usually more severe in late transition and postmenopausal periods, during which the moderate and severe symptoms were 59.1% (189/320) and 51.1% (291/570) respectively. Although most symptoms primarily appeared along with menstruation change, there are about 17.5% (172/981) patients experienced climacteric symptoms before menstruation change occurrence. There were 56.39% (733/1300) women had ever heard (mostly from gynecologist) about hormone replacement therapy from Obstetrician and Gynecologist. CONCLUSIONS: Most of the women during menopausal transition had climacteric symptoms, usually mild and moderate ones. Although most symptoms primarily appeared along with menstruation change, there are other patients' experienced climacteric symptoms before menstruation change occurrence.

[Study on relationship between endogenous androgens and insulin resistance at the different stages of postmenopause].

OBJECTIVE: To investigate the relationship between insulin resistance and endogenous androgens at early and late phase of postmenopause. METHODS: A total of 105 women with early postmenopause ( ≤ 5 years since menopause) and 107 women with late postmenopause ( ≥ 10 years since menopause) were enrolled in this study.In the mean time, those women were classified into normal weight[body mass index (BMI), BMI <24 kg/m(2)] group and overweight (BMI ≥ 24 kg/m(2)) group.Sex hormone-binding globulin (SHBG), testosterone (T) , dehydroepiandrosterone-sulfate (DHEA-S) , fasting blood glucose(FBG), fasting insulin (FINS) levels were measured and then calculated free androgen index (FAI) and homeostatic model assessment of insulin resistance (HOMA-IR) . The relationship between sex hormones and insulin resistance was analyzed by partial correlation and multiple linear regression analyses. RESULTS: Compared to early postmenopausal women, late postmenopausal women had higher FINS [(7.9 ± 6.6) mU/L versus (6.6 ± 4.0) mU/L] and HOMA-IR(2.1 ± 1.9 versus 1.7 ± 1.1), but they had lower DHEA-S[(0.9 ± 0.5) mg/L versus (1.1 ± 0.5) mg/L, all P < 0.05) ]. Both in early postmenopausal and late postmenopausal groups, overweight women had higher HOMA-IR (early group, 2.2 ± 1.0 versus 1.2 ± 0.9;late group, 2.8 ± 2.6 versus 1.6 ± 1.1) and FINS early group[(6.9 ± 2.9) mU/L versus (4.6 ± 2.0) mU/L];late group[(10.2 ± 9.3) mU/L versus (6.4 ± 3.6) mU/L] than those at women with normal weight group (all P < 0.05) .In early postmenopausal group, overweight women had lower SHBG[ (52 ± 37) nmol/L versus (71 ± 37) nmol/L] and higher FAI (2.5 ± 2.1) versus (1.3 ± 1.1) than those at normal weight women group (all P < 0.05) .In late postmenopausal group, overweight women had higher DHEA-S (1.0 ± 0.5) mg/L versus (0.8 ± 0.4) mg/L (P < 0.05) . The analyses suggested that in early postmenopausal group, SHBG was correlated negatively with FINS and HOMA-IR (ß = -0.386, P < 0.05;ß = -0.553, P < 0.05) , DHEA-S was correlated positively with FBG (ß = 0.348, P < 0.05) in early postmenopausal group.FAI was correlated positively with FBG in late postmenopausal group (ß = 0.505, P < 0.05) . CONCLUSIONS: The increased androgenic activities are associated with insulin resistance after of menopause. These correlations are different at different stages of postmenopause, which SHBG levels correlate with high risk of insulin resistance and DHEA-S levels correlates with high blood glucose levels at early postmenopause and FAI correlates with high blood glucose levels at late postmenopause.

Correction: The Relationship between Endogenous Androgens and Body Fat Distribution in Early and Late Postmenopausal Women.

[This corrects the article DOI: 10.1371/journal.pone.0058448.].

[Effects of daidzein on steroid receptor coactivator-1 expression in MC3T3-E1 cells and the mechanism].

OBJECTIVE: To investigate the roles of daidzein in the expressions of steroid receptor coactivator-1 (SRC-1) and nuclear receptor corepressor (NcoR) in MC3T3-E1 osteoblastic cells. METHODS: MC3T3-E1 cells were cultured in α-minimal essential medium (α-MEM) containing 2% fetal bovine serum and treated with various concentrations of daidzein (10(-9), 10(-7) and 10(-5) mol/L) or 17ß-estradiol at 10(-8) mol/L for 3 d. The protein levels of SRC-1 and NcoR in MC3T3-E1 cells were determined by Western blotting. Estrogen receptor (ER) antagonist ICI182780 at 10(-7) mol/L or specific ERα antagonist methyl-piperidino-pyrazole (MPP) at 10(-6) mol/L were used to block the corresponding receptors, and then MC3T3-E1 cells were treated with daidzein at 10(-7) mol/L or 10(-5) mol/L for 3 d. SRC-1 and NcoR protein levels were detected by Western blotting. RESULTS: The protein levels of SRC-1 increased by 2.5 fold (P<0.05) and 2 fold (P<0.05) by 10(-7) and 10(-5) mol/L of daidzein respectively, while the NcoR levels were not significantly altered. 17ß-Estradiol at dose of 10(-8) mol/L did not affect the expression of SRC-1 but decreased NcoR protein expression by 35% (P<0.05). Compared with the control, daidzein at 10(-7) and 10(-5) mol/L did not increase SRC-1 expression when ERs were blocked by antagonist ICI182780. Daidzein at 10(-7) and 10(-5) mol/L up-regulated SRC-1 by 1.8 fold (P<0.05) and 2.4 fold (P<0.05) respectively while ERα was blocked by MPP. CONCLUSION: Daidzein increases protein level of SRC-1 and the ratio of SRC-1/NcoR. ERß, instead of ERα, participates in the action of daidzein in regulating SRC-1 expression. Up-regulation of SRC-1 and increase of SRC-1/NcoR are part of the mechanism of the estrogenic effect of daidzein in improving osteogenesis.

Accumulation of nerve growth factor and its receptors in the uterus and dorsal root ganglia in a mouse model of adenomyosis.

BACKGROUND: Adenomyosis is a common gynecological disease, which is accompanied by a series of immunological and neuroendocrinological changes. Nerve growth factor (NGF) plays a critical role in producing pain, neural plasticity, immunocyte aggregation and release of inflammatory factors. This study aimed to investigate the expression of NGF and its two receptors in uteri and dorsal root ganglia (DRG) in an adenomyosis mouse model, as well as their relationship with the severity of adenomyosis. METHODS: Forty newborn ICR mice were randomly divided into the adenomyosis model group and control group (n = 20 in each group). Mice in the adenomyosis model group were orally dosed with 2.7 µmol/kg tamoxifen on days 2-5 after birth. Experiments were conducted to identify the expression of NGF- beta and its receptors, tyrosine kinase receptor (trkA) and p75 neurotrophin receptor (p75NTR), in the uterus and DRG in four age groups (90+/-5 d, 140+/-5 d, 190+/-5 d and 240+/-5 d; n = 5 mice in each group) by western bolt, immunochemistry and real time reverse transcription-polymerase chain reaction. RESULTS: Adenomyosis, which became more serious as age increased, was successfully induced in dosed ICR mice. NGF-beta, trkA and p75NTR protein levels in the uterus and trkA mRNA levels in DRG were higher in the older aged adenomyosis model group than those in controls (190+/-5 d and 240+/-5 d groups, P < 0.05). The expression of NGF-beta and its receptors in the uterus increased gradually as age increased for adenomyosis mice (190+/-5 d and 240+/-5 d, P < 0.05, compared with 90+/-5 d) but it showed little change in control mice. The mRNA level of trkA in DRG also increased as age increased in the adenomyosis model group (190+/-5 d and 240+/-5 d, P < 0.05, compared with 90+/-5 d) but was unchanged in controls. The mRNA level of p75NTR in DRG was not different between the adenomyosis and control groups and was stable from young to old mice. CONCLUSIONS: NGF- beta can be used as an indicator for the severity of adenomyosis. The gradually increasing level of NGF- beta and its receptors while the disease becomes more severe suggests an effect of NGF- beta on pathogenic mechanisms of adenomyosis.

[Dose-dependent effects of daidzein in regulating bone formation through estrogen receptors and peroxisome proliferator-activated receptor γ].

OBJECTIVE: To investigate different doses of daidzein (DAI) in regulating bone formation of osteoblasts, and the regulating mechanisms of estrogen receptors (ERs) and peroxisome proliferator-activated receptor γ (PPARγ) in bone formation. METHODS: Human fetal osteoblasts (hFOBs) incubated without any treatment were served as controls (control group). The hFOBs were exposed to DAI of 10(-9), 10(-7) and 10(-5) mol/L for 72 h, and to ß-estradiol-17-valerate (E(2)) of 10(-8) mol/L as positive control, respectively. Methyl thiazolyl tetrazolium assay was employed to determine the proliferation status of osteoblasts, and 4-nitrophenyl phosphate disodium salt (PNPP) method was employed to determine the activity of alkaline phosphatase (ALP). ER antagonist ICI 182780 (ICI), ERα-selective antagonist methyl-piperidino-pyrazole (MPP) and irreversible PPARγ antagonist GW9662 (GW) were used to block the corresponding receptor, while hFOBs were exposed to E(2) or different concentrations of DAI for 48 h. MTT assay and PNPP method were used respectively to determine the proliferation status and ALP activity of osteoblasts cultured in vitro. RESULTS: The osteoblast proliferation rate decreased progressively as the dose of DAI increased. Compared with the controls, the osteoblast proliferation rate in the DAI 10(-9) mol/L group increased significantly, while DAI 10(-5) mol/L group decreased significantly (P<0.05). ALP level decreased progressively as the dose of DAI increased, but there was no significant difference between groups (P>0.05). When ERs were blocked by ICI, proliferation rates in the E(2) group and DAI 10(-9), 10(-7) and 10(-5) mol/L groups were 88.16%, 76.30%, 81.18% and 83.19% respectively, which were all significantly lower than before (P<0.05). After ERα was blocked by MPP alone, proliferation rates in E(2) group and DAI 10(-9), 10(-7) and 10(-5) mol/L groups were 69.78%, 63.31%, 70.71% and 78.43%, respectively, which were also significantly lower than before (P<0.05). ALP level in the DAI 10(-9) mol/L group decreased significantly when ERα was blocked alone. When PPARγ inhibitor GW was added to the culture system, proliferation rates in E(2) group and DAI 10(-9), 10(-7) and 10(-5) mol/L groups were 103.14%, 96.99%, 112.88% and 122.22%, respectively. Compared with before, proliferation rates in DAI 10(-7) and 10(-5) mol/L groups increased significantly (P<0.05), and ALP level increased significantly (P<0.05) in the DAI 10(-5) mol/L group. CONCLUSION: DAI shows a biphasic effect on osteoporosis, whereby the effect is dose-dependent; a low-dose DAI stimulates proliferation of osteoblasts, while a high-dose DAI inhibits proliferation of osteoblasts. Low-dose DAI mainly acts on ERs, whereas high-dose DAI mainly acts on PPARγ to inhibit proliferation of osteoblasts and to some extent, acts on ERs to promote the proliferation of osteoblasts.