Increased Burden of Second Bladder Cancer and Rectal Cancer in Prostate Cancer Treated With Radiotherapy: Results From Surveillance, Epidemiology, and End Results.

BACKGROUND: Previous studies have confirmed the higher risk of bladder cancer (BC) and rectal cancer (RC) development among prostate cancer (PCa) patients receiving radiotherapy. In this study, we intend to explore the long-term trend in second BC and RC incidence among PCa patients undergoing radiotherapy. METHOD: We identified first primary PCa patients diagnosed between 1975 and 2014 from the Surveillance, Epidemiology, and End Results (SEER)-9 cancer registries. Standardized incidence ratios (SIRs) were calculated by calendar year of diagnosis among PCa patients receiving radiotherapy and not. P trends were evaluated using Poisson regression. 10-year cumulative incidence of BC and RC was calculated utilizing competing risk regression model. RESULT: Of PCa patients treated with radiotherapy, SIRs of BC increased from .82 (95% CI: .35- 1.61) in 1980-1984 to 1.58 (95% CI: 1.48-1.68) in 2010-2014 (Ptrend=.003). SIRs of RC increased from 1.01 (95% CI: .27-2.58) in 1980-1984 to 1.54 (95% CI: 1.31-1.81) in 2010-2014 (Ptrend=.025). No statistically significant change in both BC and RC incidence was observed. The 10-year cumulative incidence of BC increased from 1975-1984 (.04%) to 2005-2014 (.15%) among PCa treated with radiotherapy. Simultaneously, the 10-year cumulative incidence of RC was demonstrated to range from 1975-1984 (.02%) to 2005-2014 (.11%). CONCLUSION: we have observed an increasing trend in second BC and RC incidence in PCa patients receiving radiotherapy. There was no significant change in the incidence of second BC and RC in PCa without radiotherapy. These results reflect the increasing clinical burden of second malignant tumors in PCa patients undergoing radiotherapy.

Comprehensive analysis of the expression, prognostic value and biological importance of OVO­like proteins in clear cell renal cell carcinoma.

This study examined the expression levels of OVO-like proteins (OVOLs) in clear cell renal cell carcinoma (ccRCC) tissues and their value in predicting disease prognosis. The transcript levels, genetic alterations, and biological functions of OVOLs and their correlation with tumor immune cell infiltration and drug sensitivity and survival outcomes, as well as their prognostic values, in patients with ccRCC were analyzed based on data obtained from The Cancer Genome Atlas, Gene Expression Profiling Interactive Analysis, cBioPortal, and GSCALite databases. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed using R software (Bioconductor, clusterProfiler packages). A protein-protein interaction network was established and visualized using the R software with the ggplots package. The ggstatsplot package was used to plot the correlation between gene expression and immune cell infiltration. The mRNA expression levels of OVOL1 and OVOL2 were significantly downregulated in patients with ccRCC, whereas those of OVOL3 were upregulated. OVOL1 expression was correlated with tumor stage and histological grades. The OVOL1, OVOL2, and OVOL3 levels were significantly correlated with the prognosis of patients with ccRCC, the infiltration of immune cells, and drug sensitivity. Multivariate and univariate analyses showed that the expression of OVOL1 was an independent prognostic factor for the overall survival (OS) of patients with ccRCC. The OVOL proteins were associated with various pathways, including tight junction, cell adhesion molecules, and ether lipid metabolism. Additionally, OVOL3 upregulation, and OVOL1 and OVOL2 downregulation in clinical ccRCC samples were experimentally verified. Thus, OVOL1 and OVOL2 are potential therapeutic targets and prognostic markers for ccRCC. Additionally, OVOL1 can serve as an independent prognostic factor for OS in patients with ccRCC.

IL-34 was high in serum of women with polycystic ovary syndrome and may function as potential diagnostic biomarker and therapeutic target.

BACKGROUND: Polycystic ovary syndrome (PCOS) is one of the most prevalent endocrine disorders in females of reproductive age, with a prevalence of 20%-33% in the general population. Interleukin (IL)-34 is a recently explored proinflammatory cytokine and is an important modulator in different disease types. However, the function of IL-34 in PCOS has yet to be investigated. OBJECTIVE: The purpose of this study was to determine the IL-34 serum level in women with PCOS and to compare it to that of a relatively healthy control group. Focusing on its relationship with IL-6, TNF-α, and IL-1ß and homeostatic model assessment for insulin resistance (HOMA-IR), triglyceride, and low-density lipoprotein cholesterol (LDL-C). MATERIALS AND METHODS: In this study, blood samples were obtained from 100 women with PCOS and 100 healthy control women for the purpose of estimating their serum levels of IL-34, IL-6, TNF-α, and IL-1ß using the enzyme-linked immunosorbent assay technique. RESULTS: Serum levels of IL-34, IL-6, TNF-α, and IL-1ß were all higher in PCOS women than in healthy controls, and the difference was highly statistically significant. Serum IL-34 concentration was positively correlated with IL-6, TNF-α, and IL-1ß concentration. Additionally, serum concentrations of IL-34 were positively correlated with HOMA-IR, triglyceride, and LDL-C. CONCLUSION: When compared to normal women, IL-34, IL-6, TNF-α, and IL-1ß levels were highly statistically significant in PCOS, and these high levels were associated with other cytokines (IL-6, TNF-α, and IL-1ß), HOMA-IR, triglyceride, and LDL-C.

Effect of continuous positive airway pressure on albuminuria in patients with obstructive sleep apnea: a meta-analysis.

PURPOSE: A relationship between albuminuria and obstructive sleep apnea (OSA) has been documented in previous studies. Nevertheless, the impact of continuous positive airway pressure (CPAP) treatment on albuminuria in subjects with OSA is debated. This meta-analysis was carried out to investigate whether or not CPAP treatment affected urinary albumin-to-creatinine ratio (UACR) in subjects with OSA. METHODS: A comprehensive literature search was conducted on Web of Science, Embase, and PubMed from January 1990 to December 2020. Information on patients' characteristics, features of the studies, and UACR of pre- and post-CPAP treatment was collected. For estimation of the pooled effects, standardized mean difference (SMD) was applied. RESULTS: This meta-analysis included 6 articles and 211 subjects. The pooled analysis suggested that CPAP therapy exerted a favorable effect on the decrease of UACR in subjects with OSA (SMD = 0.415, 95% CI = 0.026 to 0.804, z = 2.09, p = 0.037). Subgroup analyses revealed that the CPAP treatment effect was not influenced by sample size, BMI, age, or AHI. CONCLUSION: The present meta-analysis indicated that UACR was significantly reduced by CPAP therapy in subjects with OSA. Further well-designed randomized controlled trials with large sample size are required to confirm the benefits.

Lack of cyclical fluctuations of endometrial GLUT4 expression in women with polycystic ovary syndrome: Evidence for direct regulation of GLUT4 by steroid hormones.

Background Determination of the role of steroid hormones in expression and regulation of endometrial glucose transport 4 (GLUT4) in humans is important for understanding endometrial disorders such as polycystic ovary syndrome (PCOS), a common hormone-imbalance disease. Methods Endometrial biopsy samples were collected from non-PCOS patients with regular menstrual cycles or with hyperplasia and from PCOS patients with or without hyperplasia. In addition, endometrial tissues from postmenopausal women were incubated with human chorionic gonadotropin (hCG, 10 IU/ml), 17ß-estradiol (E2, 10 nM), progesterone (P4, 100 nM), or a combination of E2 and P4 for 24 h. The expression of GLUT4 was measured at the mRNA level using quantitative real-time polymerase chain reaction (qRT-PCR) and at the protein level using Western blot analysis and immunohistochemistry. Results A cyclical change in GLUT4 expression pattern was observed in non-PCOS patients, and a high level of GLUT4 expression was seen in the proliferative phase compared to the secretory phase. Low levels of GLUT4 expression were found in PCOS patients compared to menstrual cycle phase-matched non-PCOS patients, and there was no significant change in GLUT4 expression in PCOS patients during the menstrual cycle. GLUT4 was localized in both epithelial and stromal cells, with notable changes in epithelial cells. We postulate that decreased GLUT4 expression might be regulated by steroid hormones. In support of this, we showed that in cultured endometrial tissues hCG and E2 alone had no effect on GLUT4 expression. However, P4 alone and P4 in combination with E2 decreased GLUT4 expression. Compared with non-PCOS controls, PCOS patients with endometrial hyperplasia exhibited decreased GLUT4 expression in particular in the epithelial cells. Conclusion We conclude that P4 can induce changes in endometrial GLUT4 expression during the menstrual cycle and that abnormal hormonal conditions such as PCOS disrupt normal patterns of GLUT4 expression in endometrial cells.

Testosterone enhances lipopolysaccharide-induced interleukin-6 and macrophage chemotactic protein-1 expression by activating the extracellular signal-regulated kinase 1/2/nuclear factor-κB signalling pathways in 3T3-L1 adipocytes.

Low-grade chronic inflammation is commonly found in patients with polycystic ovary syndrome (PCOS) who exhibit hyperandrogenism or hyperandrogenemia. Clinical studies have shown that hyperandrogenemia is closely correlated with low-grade chronic inflammation. However, the mechanism underlying this correlation remains unclear. Recent studies have suggested that adipocytes increase the production of proinflammatory mediators such as interleukin-6 (IL-6) and macrophage chemotactic protein-1 (MCP-1) when the inflammatory signal transduction cascade system is activated by external stimuli. The present study aimed to evaluate the effects of testosterone on the innate signalling and expression of proinflammatory mediators in 3T3-L1 adipocytes, which were or were not induced by lipopolysaccharide (LPS). The effects of testosterone on the expression of proinflammatory mediators, nuclear factor-κB (NF-κB), and extracellular signal-regulated kinase 1/2 (ERK1/2) signalling pathways were investigated using an enzyme-linked immunosorbent assay, reverse transcriptase-polymerase chain reaction, western blot analysis and an electrophoresis mobility shift assay. Testosterone induces IL-6 and MCP-1, and enhances LPS-induction of IL-6 and MCP-1. However, the effects are not simply additive, testosterone significantly enhanced the effects of LPS-induced inflammation factors. Testosterone induces the phosphorylation of ERK1/2 and NF-κB. The effect of testosterone on the expression of IL-6 and MCP-1 is inhibited by PD98059 , an ERK1/2 inhibitor, and PDTC, an NF-κB inhibitor. The results indicate that testosterone enhances LPS-induced IL-6 and MCP-1 expression by activating the ERK1/2/NF-κB signalling pathways in 3T3-L1 adipocytes.

Direct effects of metformin in the endometrium: a hypothetical mechanism for the treatment of women with PCOS and endometrial carcinoma.

Although a number of in vitro studies have demonstrated the antiproliferative, anti-invasive, and antimetastatic effects of metformin in multiple cancer cell types, its cellular and molecular mechanisms of anti-cancer action in the endometrium of women with polycystic ovary syndrome (PCOS) have not yet been fully elucidated. Organic cation transporters (OCTs) and multidrug and toxin extrusion proteins (MATEs) are known to be involved in metformin uptake and excretion in cells. In this article, we discuss the novel therapeutic possibilities for early-stage endometrial carcinoma (EC) in women with PCOS focusing on metformin, which might have a direct effect in the endometrium through the OCTs and MATEs. We then review the molecular mechanism(s) of the action of metformin in the endometrium and highlight possible mechanistic insights into the inhibition of cell proliferation and tumor growth and, ultimately, the reversal of early-stage EC into normal endometria in women with PCOS.

Combination of Diane-35 and Metformin to Treat Early Endometrial Carcinoma in PCOS Women with Insulin Resistance.

BACKGROUND: Young women with polycystic ovary syndrome (PCOS) have a high risk of developing endometrial carcinoma. There is a need for the development of new medical therapies that can reduce the need for surgical intervention so as to preserve the fertility of these patients. The aim of the study was to describe and discuss cases of PCOS and insulin resistance (IR) women with early endometrial carcinoma while being co-treated with Diane-35 and metformin. METHODS: Five PCOS-IR women who were scheduled for diagnosis and therapy for early endometrial carcinoma were recruited. The hospital records and endometrial pathology reports were reviewed. All patients were co-treated with Diane-35 and metformin for 6 months to reverse the endometrial carcinoma and preserve their fertility. Before, during, and after treatment, endometrial biopsies and blood samples were obtained and oral glucose tolerance tests were performed. Endometrial pathology was evaluated. Body weight (BW), body mass index (BMI), follicle-stimulating hormone (FSH), luteinizing hormone (LH), total testosterone (TT), sex hormone-binding globulin (SHBG), free androgen index (FAI), insulin area under curve (IAUC), and homeostasis model assessment of insulin resistance (HOMA-IR) were determined. RESULTS: Clinical stage 1a, low grade endometrial carcinoma was confirmed before treatment. After 6 months of co-treatment, all patients showed normal epithelia. No evidence of atypical hyperplasia or endometrial carcinoma was found. Co-treatment resulted in significant decreases in BW, BMI, TT, FAI, IAUC, and HOMA-IR in parallel with a significant increase in SHBG. There were no differences in the FSH and LH levels after co-treatment. CONCLUSIONS: Combined treatment with Diane-35 and metformin has the potential to revert the endometrial carcinoma into normal endometrial cells in PCOS-IR women. The cellular and molecular mechanisms behind this effect merit further investigation.

Endometrial progesterone resistance and PCOS.

Polycystic ovary syndrome (PCOS) is a state of altered steroid hormone production and activity. Chronic estrogen exposure or lack of progesterone due to ovarian dysfunction can result in endometrial hyperplasia and carcinoma. A key contributor to our understanding of progesterone as a critical regulator for normal uterine function has been the elucidation of progesterone receptor (PR) expression, regulation, and signaling pathways. Several human studies indicate that PR-mediated signaling pathways in the nucleus are associated with progesterone resistance in women with PCOS. The aim of this review is to provide an overview of endometrial progesterone resistance in women with PCOS; to present the PR structure, its different isoforms, and their expression in the endometrium; to illustrate the possible regulation of PR and PR-mediated signaling in progesterone resistance in women with PCOS; and to discuss current clinical treatments for atypical endometrial hyperplasia and endometrial carcinoma in women with PCOS and accompanying progesterone resistance.

Prevalence of polycystic ovary syndrome in women in China: a large community-based study.

STUDY QUESTION: What is the prevalence of polycystic ovary syndrome (PCOS) in Han Chinese women from different communities? SUMMARY ANSWER: The prevalence of PCOS in Chinese women aged 19-45 years is 5.6%. WHAT IS KNOWN ALREADY: The prevalence of PCOS is reported to range from 5 to 10% but to the best of our knowledge the Han Chinese population has not been studied. STUDY DESIGN, SIZE, DURATION: A large-scale epidemiological study was carried out between October 2007 and September 2011 in 15 924 Han Chinese women of reproductive age (19-45 years) from the 10 provinces and municipalities in China. PARTICIPANTS/MATERIALS, SETTING, METHODS: A total of 16 886 women from 152 cities and 112 villages were involved in the study. All study participants received a questionnaire and underwent a physical and transvaginal ultrasound examination. Blood samples were collected from a subsample of women (n = 3565) for analysis of metabolic markers and hormones. Based on the Rotterdam PCOS criteria, we assessed hyperandrogenism (H), chronic anovulation (O) and polycystic ovaries (P). Following diagnosis, women with PCOS were assigned to one of four different phenotypes. Finally, the prevalence and related risks of PCOS among Chinese women were estimated based on all the data sources. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 16 886 women were initially involved in the study and 15 924 eligible participants then completed the study; the overall response rate was 94.3% (15 924/16 886). The prevalence of PCOS in the Chinese community population was 5.6% (894/15 924). Blood samples were analyzed from 833 of these women who were assigned to the four PCOS phenotypes as follows: 19% H + O, 37% H + P, 15% O + P and 29% H + O + P. Comparing the 833 women with PCOS to 2732 women without PCOS indicated that PCOS occurs in younger women (P < 0.05) and these women were prone not only to menstrual problems, hyperandrogenism, PCO and infertility but also metabolic syndrome (MS) and insulin resistance (IR). However, there was no significant difference in the rate of hypertension or hyperlipemia between the two groups. Obese patients with PCOS had a higher rate of MS (16 versus 48%), IR (7 versus 28%), hypertension (8 versus 30%) and hyperlipemia (48 versus 73%) compared with non-obese patients (all P < 0.05), respectively. The rates of metabolic complications in patients with PCOS increased with age. LIMITATIONS, REASONS FOR CAUTION: Age and ethnic origin contribute to the differing manifestations of PCOS; therefore, sampling is one of the most important issues in epidemiological research into PCOS. Owing to the mobility of the Chinese population, the survey among resident populations caused a certain deviation in the age distribution. WIDER IMPLICATIONS OF THE FINDINGS: The prevention and treatment of PCOS, particularly in those who are obese, are essential in Chinese women of reproductive age.

[Effects of testosterone on insulin sensitivity in liver cells and related molecular mechanisms].

OBJECTIVE: To explore the regulation of insulin sensitivity in liver cells by androgen signaling. METHODS: Eleven adult female C57BL/6 mice were injected daily with testosterone (group T) for 24 weeks. And 10 control mice received sesame oil only (group Con). HepG2 liver cells were initially pretreated with different doses of testosterone (10(-9)-10(-5) mol/L ) for 0-36 h or with 10(-7) mol/L testosterone for 0-96 h followed by a stimulation of 100 nmol/L insulin for 15 min. Later HepG2 cells were pretreated with 10(-7) mol/L testosterone for 36 h followed by a stimulation of 100 nmol/L insulin for 15 min and then a restimulation of 100 nmol/L insulin for 15 min at 4 h and 6 h interval respectively. Phosphorylation and protein expression of Akt and GSK3ß in C57BL/6 mice liver tissues and HepG2 cells were analyzed by Western blot. RESULTS: The 24-week treatment of testosterone decreased the phosphorylation of Akt and GSK3ß in C57BL/6 adipose and liver tissues (43.1% ± 3.2% vs 77.1% ± 6.7%, 14.7% ± 6.7% vs 82.3% ± 2.0% respectively, P < 0.05). Pretreatment with 10(-8)-10(-6) mol/L testosterone within 36 h obviously increased the phosphorylation of Akt and GSK3ß (P < 0.05). However pretreatment with 10(-5) mol/L within 36 h or with 10(-7) mol/L for 96 h had no effect on the phosphorylation of Akt and GSK3ß compared with control group (P > 0.05). Pretreatment with 10(-7) mol/L testosterone for 36 h followed by insulin stimulation and restimulation after 6 h interval obviously decreased the phosphorylation of Akt and GSK3ß (P < 0.05). CONCLUSION: Androgen signaling may contribute to insulin resistance in liver cells.

[Ovarian ultrasonographic features in reproductive age females with polycystic ovary syndrome and the establishment of ultrasonographic criteria].

OBJECTIVE: To understand the ovarian ultrasound imaging features in the reproductive age females with polycystic ovary syndrome (PCOS). METHODS: A total of 396 PCOS patients aged 18 - 35 years were recruited from our gynaecology & endocrinology clinic, including obese (OB-PCOS group, n = 153) and non-obese (NOB-PCOS group, n = 241). And 635 reproductive period females with normal menstruation for the control group, including obese (OB-CON group, n = 72) and non-obese (NOB-CON group, n = 563). Questionnaire surveys were conducted on their menstrual history. Vagina or rectum ultrasound methods were employed to determine the values of ovarian follicle number (FN) and ovarian volume (OV). Also the clinical symptom scores and endocrine and metabolic indices were measured. RESULTS: (1) As compared to the control group, the values of ovarian FN and OV 95% site for physiological high limit were 10 and 9.5 ml respectively. (2) In PCOS patients, their ovarian volumes and the number of follicles were significantly higher than those of the control group (P < 0.01). (3) In 90.4% of these patients, their values were OV > 9.5 ml and/or FN ≥ 10. And in 66.9% PCOS patients of reproductive age, the polycystic ovarian changes of ultrasound imaging reached the Rotterdam consensus diagnostic criteria. CONCLUSION: The features of ovarian ultrasound imaging in reproductive period PCOS patients are enlarged ovarian volume and increased follicles numbers. The preliminary Chinese ovarian ultrasonographic diagnostic cut-off points have been proposed for reproductive period PCOS patients.

[Evaluation the efficacy and safety of estradiol and drospirenone tablets in the treatment of menopausal symptoms among postmenopausal Chinese healthy women:a randomized, multi-center, double-blind, placebo-controlled clinical study].

OBJECTIVE: To study the efficacy and safety of estradiol and drospirenone tablets (Angeliq) in treatment of menopausal symptoms among postmenopausal Chinese healthy women. METHODS: Total 244 postmenopausal Chinese healthy women who had moderate to severe hot flushes were randomly assigned into estradiol and drospirenone (observation group, n = 183) or placebo group (n = 61) by the ratio of 3:1 for 16 weeks in this randomized multi-center double-blind placebo-controlled study. During the trial, the follow-up visits were conducted at week 4, 8, 12, 16 of treatment and 2 weeks after treatment respectively. Height, weight, vital signs, hot flushes, other relevant menopausal symptoms and vaginal bleeding were observed in each follow-up visit, while the clinical global impression scale was assessed at 16 weeks as well. RESULTS: It showed that hot flushes were reduced significantly more in observation group than that in placebo group (P < 0.01), although both treatments were effective. The absolute values of mean severity index of total hot flushes decreased by -0.6 ± 0.5 in observation group and -0.4 ± 0.4 in placebo group from baseline respectively, which reached significant difference (P < 0.05). However, the absolute values of mean severity index of moderate to severe hot flushes decreased by -0.6 ± 0.8 in observation group and -0.3 ± 0.6 in placebo group from baseline respectively, which had no significant difference (P > 0.05). After 16 weeks treatment, it also showed that estradiol and drospirenone had significant better efficacy than placebo on moderate to severe sweating, vaginal dryness and clinical global impression scale (P < 0.01). During the trial, blood pressure in observation group was stable. The rate of vaginal bleeding in observation group was higher than that in the placebo group, especially during the week 4 to week 8 when 48.9% (87/178) in observation group and 10.7% (6/56) in placebo group of patients bled. Although the cumulative amenorrhea rate of observation group was lower than that of placebo group in each cycle (28 days), it increased gradually along with duration of the treatment. The commonest adverse event in observation group was breast tenderness which accounted for 12.0% (22/183). The level of serum potassium was in the normal range in observation group mostly.Meanwhile, the other adverse events rate was low. Serious adverse events reported in this trial were assessed as not study drug related or as unlikely study drug related. CONCLUSION: Estradiol and drospirenone tablets which could effectively alleviate menopausal symptoms in postmenopausal Chinese healthy women is a novel hormone replacement therapy regimen with high safety and efficacy.

[Clinical features, reproductive endocrine and metabolic abnormalities of adolescent girls with polycystic ovary syndrome].

OBJECTIVE: To investigate the clinical features, reproductive endocrine and metabolic abnormalities in adolescent girls with polycystic ovary syndrome (PCOS). METHODS: A total of 325 adolescent girls with normal menstruation, 18 obese (OB-CON) and 307 non-obese (NOB-CON), were enrolled as controls from multiple middle schools in Shanghai, China. A total of 167 adolescent girls with PCOS, 90 obese (OB-PCOS) and 77 non-obese (NOB-PCOS), were also recruited. All cases were evaluated for their clinical manifestations, reproductive endocrine and metabolic parameters. Hyperandrogenism was determined by serum testosterone (T), free androgen index (FAI) and dehydroepiandrosterone sulfate (DHEA-S). Insulin sensitivity was measured by fasting insulin (FINS) and homeostasis model assessment of insulin resistance (HOMA-IR). RESULTS: (1) Menarche was significantly earlier in adolescent PCOS than in controls. The incidence of obesity was significantly higher in adolescent PCOS than that in control group. (2) T, FAI and DHEA-S were significantly higher in adolescent PCOS group than those in control group. FAI was higher in OB-PCOS group than in NOB-PCOS group. LH and LH/FSH were higher in PCOS groups than those in controls. LH and LH/FSH were also much higher in NOB-PCOS group than those in OB-PCOS group. (3) HOMA-IR and FINS were significantly higher in PCOS group than those in control group. Incidence of acanthosis nigricans, FINS, HOMA-IR and triglyceride were significantly higher in OB-PCOS group than those in NOB-PCOS group. (4) 95.21% PCOS girls presented with an ultrasonic morphological evidence of polycystic ovarian. CONCLUSION: The essential features of adolescent PCOS are an earlier onset of menarche, a persistent menstrual disorder over 2 years after menarche, a higher incidence of obesity, marked hyperandrogenism and insulin resistance and disorderly gonadotropin secretion in comparison with control subjects. Hyperandrogenism and insulin resistance are much more severe in obese adolescent PCOS.

[The effects of testosterone on the expression of inflammatory factors in 3T3-L1 adipocytes and its mechanism].

OBJECTIVE: To explore the effects of androgen upon the production of inflammatory Cultured factors in 3T3-L1 adipocytes and to investigate the mechanism at the molecular level. METHODS: pre-adipocytes from 3T3-L1 cell line were induced to differentiate into adipocytes. Mature adipocytes were treated with testosterone at a concentration of 10(-9) to 10(-15) mol/L for either short (0 to 30 min) or long (12, 24 and 48 h) treatment course. Secretion of inflammatory factors, i.e., interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1), in the culture medium were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of IL-6 and MCP-1 were determined by reverse transcriptase PCR. Phosphorylation of NF-KB was analyzed by Western blot with beta-actin as the reference gene. In another experiment, adipocytes were manipulated according to the same protocol except being pretreated with PDTC (inhibitor of NF-KB) for 2 h prior to the addition of testosterone. The results of two experiments were compared. RESULTS: (1) The secretion of IL-6 and MCP-1 in the culture medium were higher in the testosterone-treated groups than in the control groups (P < 0.05). The highest concentration of IL-6 and MCP-1 were observed in the group treated with 10(-5) M testosterone for 24 h. The mRNA expression of IL-6 and MCP-1 were higher in the groups treated with testosterone for 12 h, especially with testosterone of 10(-5) mol/L; (2) With a short treatment course of testosterone, more NF-kappaB were phosphorylated than in control, especially with testosterone of 10(-5) mol/L. More NF-kappaB was phosphorylated following the 12 h testosterone treatment (10(-9), 10(-7) and 10(-5) mol/L), especially with a testosterone concentration of 10(-9) and 10(-5) mol/L. (3) When pretreated by NF-kappaB inhibitor and followed by 10(-5) mol/L testosterone for 24 h, the secretion of IL-6 and MCP-1 in the culture medium decreased significantly (P < 0.01). Likewise, when pretreated by NF-kappaB inhibitor and followed by 10(-5) M testosterone for 12 h, the mRNA expression of IL-6 and MCP-1 decreased significantly. CONCLUSION: Within the certain scope, testosterone could increase the expression of inflammatory factors in adipocytes through the activation of NF-kappaB.

[Role of testosterone in tumor necrosis factor-alpha and monocyte chemotactic protein-1 expression in mouse macrophage RAW264.7 and its molecular mechanism].

OBJECTIVE: To investigate the role of androgen in TNF-alpha and MCP-1 expression in RAW264.7 macrophage and its molecular mechanism. METHODS: (1) RAW264.7 macrophage was treated with 10(-9) mol/L or 10(-7) mol/L testosterone (T) and then subjected to the measurement of: 1) the cellular expression of TNF-alpha and MCP-1 mRNA by RT-PCR; 2) the expression of TNF-alpha and MCP-1 in cell culture supernatant by ELISA; (2) The expression of phospho-NF-kappaB after treatment with T was measured by Western blot. (3) Cells were pre-incubated with 10(-4) mol/L PDTC (an inhibitor of NF-kappaB) for 1 hour, followed by T treatment. Expression of mRNA and supernatant levels of TNF-alpha and MCP-1 were measured by RT-PCR and ELISA. RESULTS: (1) 1) After a 6 h treatment with 10(-9) mol/L or 10(-7) mol/L T, the expression of TNF-alpha mRNA increased by 1.78 and 1.87 folds, MCP-1 by 1.58 and 1.66 folds respectively (all P < 0.05). 2) Incubated with both concentration of T for 6 h showed no significant changes of supernatant levels of TNF-alpha and MCP-1. After a 24 h treatment, the levels of TNF-alpha and MCP-1 increased significantly (all P < 0.05) while more significant increase was found in 10(-7) mol/L T group (P < 0.05). (2) The expression of phospho-NF-kappaB (ser276) increased significantly after cells were treated with 10(-7) mol/L T for 30 min (P < 0.05) and peaked at 60 min. (3) With 1 h PDTC pre-incubation, T (10(-9) mol/L or 10(-7) mol/L) treatment for 6 h led to a lower mRNA expression and 24 h led to lower supernatant levels of TNF-alpha and MCP-1 than those without (P < 0.05). However, both cellular and supernatant expression of TNF-alpha and MCP-1 with PDTC pre-incubation were still higher than those of blank controls (all P < 0.05, except for TNF-alpha in 10(-9) mol/L T treatment). CONCLUSION: Testosterone can increase TNF-alpha and MCP-1 expression in RAW264.7 macrophage in vitro. Activation of cellular NF-kappaB by testosterone may be one of underlying molecular mechanisms.

[Correlations between adipocytokines and insulin resistance in women with polycystic ovary syndrome].

OBJECTIVE: To investigate the correlations between adipocytokines and insulin resistance in women with polycystic ovary syndrome. METHODS: Sixty women with polycystic ovary syndrome aged 19-34 years old were divided into 2 groups: group A [n = 36, BMI > or = 25 kg/m(-2) or WHR (waist height ratio) > 0.85] and group B (n = 24, BMI < 25 kg/m(-2) and WHR < or = 0.85). Twenty-six healthy infertile women with a mean age of 26 + or - 8 years old served as controls and they were named as group C (n = 26, BMI < 25 kg/m(-2) and WHR < or = 0.85). Anthropometric measurements, hormonal profiles and metabolic profiles were compared between three groups. Plasma leptin, CRP and free fatty acid were measured by enzyme-linked immunosorbent assay (ELISA). Adiponectin, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were measured by radioimmunoassay. Insulin resistance was estimated by homeostasis model assessment (HOMA). RESULTS: (1) Adiponectin and TNF-alpha levels were significantly higher in group B (17 + or - 9) mg/L and (1.0 + or - 0.3) mg/L respectively than in group A (10 + or - 7) mg/L and (0.8 + or - 0.2) mg/L respectively. Leptin was lower in group B (24 + or - 13) microg/L and group C than in group A (42 + or - 21) microg/L (all P < 0.01). IL-6 was higher in group A (173 + or - 184) ng/L and group B (184 + or - 44) ng/L than in group C (P < 0.05 and P < 0.01 respectively). No significant difference of circulating level of CRP was found between these 3 groups. (2) Leptin was positively correlated with BMI, WHR and HOMA-IR (all P < 0.01) whereas adiponectin was negatively correlated with BMI, WHR and HOMA-IR (P < 0.01; P < 0.01, P < 0.05 respectively). Multivariate regression showed that adiponectin was the most significant predictor of HOMA-IR and it explained 16.5% of variance. CONCLUSION: The higher levels of leptin and TNF-alpha, the lower level of adiponectin in obese PCOS and the higher level of TNF-alpha and IL-6 in non-obese PCOS suggested different adipocytokines play different roles of insulin resistance in PCOS.

[Characteristics of quantitative change relation between insulin and testosterone in polycystic ovary syndrome].

OBJECTIVE: To investigate the relationship of dynamic quantitative changes between insulin and testosterone in polycystic ovary syndrome (PCOS). METHODS: Peripheral blood samples were collected in the third to fifth day menstrual cycle from 97 PCOS patients, aged (24 +/- 6), 47 being obese (with BMI > 25 kg/m(2)) and 50 being non-obese (with BMI

[Testosterone impairs insulin sensitivity in 3T3-L1 adipocytes and the related nongenomic mechanisms].

OBJECTIVE: To investigate the influence of rapid nongenomic effect of androgen on the insulin sensitivity of mature adipocytes and the molecular mechanism thereof. METHODS: Fetal mice [corrected] preadipocytes of the line 3T3-L1 were cultured to develop into mature adipocytes. 3T3-L1 adipocytes were pretreated with testosterone of the concentration of 10-9 to approximately 10(-5) mol/L for a short-time 0-30 minutes) or a long-time (24 hours). Insulin (Ins) 200 microl at the concentration 100 micromol/L and 2-deoxy [3H] glucose were added to examine the glucose uptake. Phosphorylation and protein expression of Ins receptor (InsR) and its downstream signaling molecules (Akt and GSK3beta) were analyzed by Western blotting. RESULTS: The Ins-stimulated glucose uptake after the pretreatment of testosterone for 30 min and 24 h decreased gradually in response to the increasing of the concentration of testosterone with the nadir both at the testosterone concentration of 10(-5) mol/L (both P < 0.05). The phosphorylation levels of InsR, Akt, and GSK3Pbeta were significantly down-regulated by adding of testosterone at the concentration of 10(-6) mol/L for 3-30 minutes (all P < 0.05) or by adding of testosterone at the concentrations of 10(-7) - 10(-6) mol/L for 24 hours (all P < 0.05). The protein expression levels of InsR, Akt, and GSK3Pbeta, however, were not significantly affected by testosterone treatment. CONCLUSION: Rapid nongenomic effect of androgen may contribute to the insulin resistance in adipocytes.