The Role of miRNAs 340-5p, 92a-3p, and 381-3p in Patients with Endometriosis: A Plasma and Mesenchymal Stem-Like Cell Study.

BACKGROUND: Endometriosis is the most prevalent gynecological disease with elusive etiology. The mysterious entity and the lack of noninvasive diagnostic methods affect women's lives negatively. This study is aimed at finding the relationship between miR-340-5p, 92a-3p, and miR-381-3p and the pathogenesis of endometriosis in endometrial mesenchymal stem-like cells (eMSCs) of endometriosis and assessing their potential as a noninvasive biomarker in plasma. METHODS: Peripheral blood and eMSC specimens were collected from suspected women of endometriosis before laparoscopy. Total RNA was isolated from plasma and cultured eMSCs to synthesize complementary DNA. The expression of miR-340-5p, miR-92a-3p, and miR-381-3p was analyzed by RT-qPCR. To understand these miRNAs' role, we also did a bioinformatic analysis. RESULTS: There was a downregulation of miR-340-5p, miR-92a-3p, and miR-381-3p in plasma, and the upregulation of miR-340-5p and the downregulation of miR-92a-3p and miR-381-3p in eMSCs of women with endometriosis. There was a positive concordance between the expression of miR-92a-3p and miR-381-3p in plasma and eMSCs. Our study also showed three genes, Solute Carrier Family 6 Member 8 (SLC6A8), Zinc Finger Protein 264 (ZNF264), and mouse double minute 2 (MDM2), as common targets of these miRNAs. CONCLUSIONS: This study has been one of the first attempts to examine the expression of miR-340-5p, miR-92a-3p, and miR-381-3p in both plasma and eMSCs and revealed their possible role in endometriosis based on in silico analysis. Biomarkers pave the way to develop a new therapeutic approach to the management or treatment of endometriosis patients. Our result as a first report shows that combined levels of miRNAs 340-5p and 381-3p may have the potential to be utilized as diagnostic biomarkers for endometriosis.

Combination of Ferulic Acid, Ligustrazine and Tetrahydropalmatine attenuates Epithelial-mesenchymal Transformation via Wnt/ß-catenin Pathway in Endometriosis.

Previously the potential therapeutic action of ferulic acid, ligustrazine and tetrahydropalmatine (FLT) are discovered with unclear mechanism in rat autograft endometriosis. However, the effect of FLT on endometrial cells and allograft endometriosis is still unclear. This study is designed to elucidate the influence of FLT on epithelial-mesenchymal transformation in allograft endometriosis and endometrium cells. In vivo, fluorescent xenogeneic endometriosis model was established. In vitro, invasion and metastasis were analyzed after treating FLT. Epithelial-mesenchymal transformation and Wnt/ß-catenin pathway were inspected in vitro and in vivo. Activator or inhibitor of Wnt/ß-catenin signaling was performed to inspect mechanism of epithelial-mesenchymal transformation. In vivo, FLT not only decreased fluorescent intensity and volume of ectopic lesion, but also ameliorated pathological morphology. E2 and PROG levels in serum were reduced by FLT. In endometrial cells, FLT significantly inhibited the invasion and metastasis. Meantime, epithelial-mesenchymal transformation was reversed, accompanied by suppression of Wnt/ß-catenin pathway. In-depth study, activation of Wnt/ß-catenin pathway lead to promotion of epithelial-mesenchymal transformation, which was reversed by FLT. FLT prevented fluorescent allograft endometriosis and endometrium cells, which was related to suppress epithelial-mesenchymal transformation through inactivating Wnt/ß-catenin pathway. The findings disclose molecular mechanism of epithelial-mesenchymal transformation in endometriosis by FLT, and contribute to further application.

Protopanaxadiol improves endometriosis associated infertility and miscarriage in sex hormones receptors-dependent and independent manners.

Background: Patients with endometriosis (EMs) have high risks of infertility and spontaneous abortion. How to remodel the fertility of patients with EMs has always been the hot spot and difficulty in the field of reproductive medicine. As an aglycone of ginsenosides, protopanaxadiol (PPD) possesses pleiotropic biological functions and has high medicinal values. We aimed to investigate the effect and potential mechanism of PPD in the treatment of EMs-associated infertility and spontaneous abortion. Methods: The EMs mice models were constructed by allotransplantation. The pregnancy rates, embryo implantation numbers and embryo resorption rates of control and EMs were counted. RNA sequencing, qRT-PCR, enzyme linked immunosorbent assay (ELISA) and FCM analysis were performed to screen and confirm the expression of endometrial receptivity/decidualization-related molecules, inflammation cytokines and NK cell function-related molecules in vitro and/or in vivo. The SWISS Target Prediction, STRING and Cytoscape were carried out to predict the potential cellular sensory proteins, the protein-protein interaction (PPI) network between sensory proteins and fertility-related molecules, respectively. Micro-CT detection, liver and kidney function tests were used to evaluate the safety. Results: Here, we observe that PPD significantly up-regulates endometrial receptivity-related molecules (e.g., Lif, Igfbp1, Mmps, collagens) and restricts pelvic inflammatory response (low levels of IL-12 and IFN-γ) of macrophage, and further remodel and improve the fertility of EMs mice. Additionally, PPD increases the expression of decidualization-related genes and Collagens, and promotes the proliferation, residence, immune tolerance and anagogic functions of decidual NK cells (low levels of CD16 and NKp30, high levels of Ki67, VEGF, TGF-ß) in pregnant EMs mice, and further triggers decidualization, decidual NK cell-mediated maternal-fetal immune tolerance and angiogenesis, preventing pregnant EMs mice from miscarriage. Mechanically, these effects should be dependent on ESRs, PGR and other sensory proteins (e.g., AR). Compared with GnRHa (the clinic first-line drug for EMs), PPD does not lead to the decline of serum estrogen and bone loss. Conclusion: These data suggest that PPD prevents EMs-associated infertility and miscarriage in sex hormones receptors-dependent and independent manners possibly, and provides a potential therapeutic strategy with high efficiency and low side effects to remodels the fertility of patients with EMs.

The potentialities of oxytocin receptor inhibitors for endometriosis therapy.

OBJECTIVE: Genital endometriosis (GE) is a widespread gynecological disease which requires its further pathogenesis investigation and search for new effective treatments. The known data of oxytocin receptor presence in endometrioid heterotopy smooth muscle cells give some grounds to assume oxytocin participation in the pathogenesis of endometriosis. The present study objective was to evaluate oxytocin level in peripheral blood (PB) in patients with endometriosis associated pain syndrome and to estimate the efficacy of oxytocin receptor inhibitors (IOXTR) administration based on animal endometriosis model. MATERIALS AND METHODS: The basic group comprised 61 patients with endometriosis associated pain syndrome, while 21 patients formed the control group. VAS, MPQ, and BBS objective tests were applied for pain syndrome evaluation. Oxytocin level in PB was measured by immunoenzyme method. After confirmation of endometriosis experimental model formation in rats and further randomization, a daily IOXTR intra-abdominal injection was performed in a dose of 0.35 mg/kg/24 h in the basic group (n = 12) or saline solution administration in the control (n = 12). On the final stage, endometrioid heterotopy size measuring was performed along with histological examination. RESULTS: Oxytocin level in PB was authentically higher in patients with GE compared to the control: 51.45 (35.54-62.76) pg/mL and 27.64 (23.23-34.12) pg/mL, respectively (p<.001). Positive correlation between oxytocin PB level and pain syndrome expression was established in patients with GE: VAS (r = 0.76; p<.001), MPQ (r = 0.52; p<.001), and BBS (r = 0.57; p<.001). Based on the experimental disease model authentical decrease of endometrioid heterotopy average area was observed after IOXTR therapy compared to the control (7.3 ± 1.8 mm2 and 22.2 ± 1.2 mm2, respectively, p<.05). CONCLUSIONS: The obtained results confirm the oxytocin role in the pathogenesis of endometrioid associated pain syndrome. The high efficacy of IOXTR administration based on animal model of surgically induced endometriosis allows viewing this method as a perspective therapy.

Phytochemical profiling and ameliorative effects of Achillea cretica L. on rat model of endometriosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Achillea cretica (AC) is a medicinal plant emphasized for treatment of gynecological disorders and pathological symptoms similar to endometriosis in traditional Persian medicine. Since information about its chemical constituents is limited, the aim of this study is to investigate phenolic composition of AC extract as well as its effect on experimental model of endometriosis. MATERIALS AND METHODS: RP-HPLC-PDA-ESI-MS/MS analysis was used for the determination of polyphenolic compounds. Endometriosis was induced in rats by suturing of uterus segments to abdominal wall of same rat, after eight weeks when the model was induced, it was followed by 28 days of treatment with 100, 200, and 400 mg/kg/day of hydroethanolic extract of the plant. Blood samples and implanted tissues were collected in the final day, and area of foci, tumor necrosis factor-α, vascular endothelial growth factor, interleukin-6, and serum total thiol molecules were measured and compared with positive group (0.2 mg/kg/day letrozole) and control group (solvent of extract: normal saline). Implanted tissue sections of the sacrificed rats were also assessed histopathologically. RESULTS: Nine polyphenolic compounds were identified in AC extract including 7 flavonoids and 2 phenolic acids. Plant extract decreased area of foci and cytokine levels in serum and local tissue. Histopathological assessments confirmed the effectiveness of treatments by decreasing the thickness of epithelial layer and increasing the infiltration of leukocytes into this layer. Doses of 100 mg/kg and 400 mg/kg of extract showed better effects in comparison with the dose of 200 mg/kg in reduction of cytokine levels and size of implanted tissue. Extract and letrozole did not demonstrate significant effect on thiol level. CONCLUSION: AC aerial extract may be a favorable medicine for management of endometriosis by modulating inflammatory cytokines; however, further studies are needed for more conclusive and reliable decision about its efficacy and safety.

B lymphocytes inactivation by Ibrutinib limits endometriosis progression in mice.

STUDY QUESTION: What are the effects of B lymphocyte inactivation or depletion on the progression of endometriosis? SUMMARY ANSWER: Skewing activated B cells toward regulatory B cells (Bregs) by Bruton's tyrosine kinase (Btk) inhibition using Ibrutinib prevents endometriosis progression in mice while B cell depletion using an anti-CD20 antibody has no effect. WHAT IS KNOWN ALREADY: A polyclonal activation of B cells and the presence of anti-endometrial autoantibodies have been described in a large proportion of women with endometriosis though their exact role in the disease mechanisms remains unclear. STUDY DESIGN, SIZE, DURATION: This study included comparison of endometriosis progression for 21 days in control mice versus animals treated with the anti-CD20 depleting antibody or with the Btk inhibitor Ibrutinib that prevents B cell activation. PARTICIPANTS/MATERIALS, SETTING, METHODS: After syngeneic endometrial transplantation, murine endometriotic lesions were compared between treated and control mice using volume, weight, ultrasonography, histology and target genes expression in lesions. Phenotyping of activated and regulatory B cells, T lymphocytes and macrophages was performed by flow cytometry on isolated spleen and peritoneal cells. Cytokines were assayed by ELISA. MAIN RESULTS AND THE ROLE OF CHANCE: Btk inhibitor Ibrutinib prevented lesion growth, reduced mRNA expression of cyclooxygenase-2, alpha smooth muscle actin and type I collagen in the lesions and skewed activated B cells toward Bregs in the spleen and peritoneal cavity of mice with endometriosis. In addition, the number of M2 macrophages decreased in the peritoneal cavity of Ibrutinib-treated mice compared to anti-CD20 and control mice. Depletion of B cells using an anti-CD20 antibody had no effect on activity and growth of endometriotic lesions and neither on the macrophages, compared to control mice. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: It is still unclear whether B cell depletion by the anti-CD20 or inactivation by Ibrutinib can prevent establishment and/or progression of endometriosis in humans. WIDER IMPLICATIONS OF THE FINDINGS: Further investigation may contribute to clarifying the role of B cell subsets in human endometriosis. STUDY FUNDING/COMPETING INTEREST(S): This research was supported by a grant of Institut National de la Santé et de la Recherche Médicale and Paris Descartes University. None of the authors has any conflict of interest to disclose.

Downregulation of lncrna uca1 as a diagnostic and prognostic biomarker for ovarian endometriosis

SUMMARY OBJECTIVE: Ovarian endometriosis seriously affects the quality of life of females, and long non-coding RNA lncRNA urothelial carcinoma-associated 1 (UCA1) plays pivotal roles in the pathogenesis of various ovarian diseases. However, the involvement of lncRNA UCA1 in ovarian endometriosis remains unknown to date. Therefore, the present study aims to study the role of UCA1 in ovarian endometriosis. METHODS: A total of 98 patients with ovarian endometriosis and 28 healthy females were included. The expression of lncRNA UCA1 in ectopic and eutopic endometrium tissues of ovarian endometriosis patients and controls was detected using qRT-PCR. A ROC curve analysis was performed to evaluate the diagnostic values of serum lncRNA UCA1 for ovarian endometriosis. Patients were followed up for 2 years after discharge, and the recurrence of ovarian endometriosis was recorded. RESULTS: The expression level of lncRNA UCA1 was significantly higher in ectopic endometrium tissues than in paired eutopic endometrium tissues for most of the patients. The serum lncRNA UCA1 level showed no significant correlations with either patients' age or living habits. After the treatment, the serum lncRNA UCA1 level increased, and serum levels of lncRNA UCA1 on the day of discharge were significantly lower in patients with recurrence than those in patients without recurrence. Conclusion: The downregulation of lncRNA UCA1 is involved in the pathogenesis of ovarian endometriosis and may serve as a promising diagnostic and prognostic biomarker for the disease.

RESUMO OBJETIVO: A endometriose ovariana afeta seriamente a qualidade de vida das mulheres, e o carcinoma urotelial 1 de urcélio de RNA não codificador longo 1 (UCA1) desempenha um papel crucial na patogênese de várias doenças ovarianas. No entanto, o envolvimento do lncRNA UCA1 na endometriose ovariana permanece desconhecido até o momento. Portanto, o presente estudo tem como objetivo estudar o papel do UCA1 na endometriose ovariana. Métodos: Um total de 98 pacientes com endometriose ovariana e de 28 mulheres saudáveis foi incluído. A expressão de lncRNA UCA1 em tecidos de endométrio ectópico e eutópico de pacientes com endometriose ovariana e controles foi detectada por qRT-PCR. A análise da curva ROC foi realizada para avaliar os valores diagnósticos do lncRNA UCA1 sérico para endometriose ovariana. Os pacientes foram acompanhados por dois anos após a alta, e a recorrência da endometriose ovariana foi registrada. RESULTADOS: O nível de expressão do lncRNA O UCA1 foi significativamente maior nos tecidos do endométrio ectópico do que nos tecidos do endométrio eutópico pareados para a maioria dos pacientes. O nível sérico de UCA1 foi diminuído com a progressão da endometriose ovariana. O soro UCA1 pode ser usado para diagnosticar com precisão a endometriose ovariana. O nível sérico de UCA1 não apresentou correlações significativas com a idade ou com os hábitos de vida dos pacientes. Após o tratamento, o nível sérico do lncRNA UCA1 foi aumentado, e os níveis séricos de lncRNA UCA1 no dia da alta foram significativamente menores nos pacientes com recidiva do que naqueles sem recorrência. CONCLUSÃO: A regulação negativa do lncRNA UCA1 está envolvida na patogênese da endometriose ovariana e pode servir como um promissor biomarcador diagnóstico e prognóstico para a doença.

Evaluation of serum cancer antigen 125, resistin, leptin, homocysteine, and total antioxidant capacity in rat model of endometriosis treated with Curcumin.

Endometriosis is one of the most common reproductive diseases of women, with some important biochemical changes in the serum. Curcumin was reported to have anti-endometriosis, anti-inflammatory and antioxidant activity. This study explores the changes of serum levels of Cancer Antigen 125 (CA125), leptin, resistin, homocysteine, and total antioxidant capacity (TAC) in a rat model of endometriosis and the effect of curcumin treatment on these factors. Fifty female Sprague-Dawley rats (220-250 gr body weight) were randomly divided into control (received 0.3 mL of the vehicle), sham(stress of surgery + 0.3 mL of the vehicle) and three endometriosis groups as danazol treatment (7.2 mg/kg BW, IP), curcumin treatment (48 mg/kg BW, IP) and test (received 0.3 mL of the vehicle). Endometriosis was induced by surgically auto-transplantation of uterine tissue to the abdominal wall and intestinal mesentery. The animals were treated for 4 weeks. On the last day, the blood sample was collected by heart puncture, and the above-mentioned factors were measured in the sera. Leptin in the curcumin treatment group was markedly higher than all of the studied groups, except the danazol group, while there were no significant differences between other groups (P < 0.05). Level of resistin in endometriosis groups did not show significant differences with the control group (P < 0.05). There was no significant difference in the serum levels of homocysteine and CA 125 between all of the studied groups (P > 0.05). The serum levels of TAC in the control group were significantly higher than all of the studied groups (P < 0.05). Curcumin could prevent the growth of endometriosis, while there was no significant alteration on measured factors in the serum of rat with induced endometriosis. Hence follow up of these parameters in serum may not be a proper indicator to evaluate the status of endometriosis in the rat model.

Heavy black tea consumption and elevated CA 19-9 and CA 125 levels. A case report on a patient with ovarian endometriotic cysts.

In this case report, we present a case of false positive CA 19-9 and CA 125 levels in a patient with suspected endometriotic cysts. The patient is a 34-year-old nulliparous woman with heavy black tea consumption and elevated CA 19-9 and CA 125 levels. After discontinuation of black tea intake and careful exploration of other possible factors, CA 19-9 and CA 125 levels dropped markedly. As a conclusion, heavy black tea consumption can lead to false positive results of elevated CA 19-9 and CA 125 levels.

Elevated serum chemokines are independently associated with both endometriosis and uranium exposure.

Endometriosis is a complex disease impacted by the hormonal and immune systems. Cytokines and chemokines are serum biomarkers that maybe useful to develop a noninvasive disease diagnosis. Individuals in the Fernald Community Cohort were exposed to uranium, a heavy metal with radioactive properties and estrogenic potential; therefore, serum samples from women in this cohort with or without uranium and with or without endometriosis were compared for alterations in chemokine, cytokine, and matrix metalloproteinase (MMP) levels. Control women were matched to endometriosis cases by uranium exposure, age, and body mass index. MMP levels were not altered. Five chemokines and one cytokine significantly increased in endometriosis cases versus controls irrespective of uranium exposure. Uranium exposure alone was associated with an increase in inflammatory chemokines. The majority of the elevated chemokines in endometriosis cases play important roles in attracting T helper-2 cells, which may be vital to understanding the immune response in endometriosis.

Differential Proteomics Analysis of Endometriosis in Blood Stasis Syndrome.

OBJECTIVE: To investigate the innate characters of 3 endometriosis (EMT) syndromes, blood stasis (BS), qi stagnation and blood stasis (QSBS) as well as Shen (Kidney) deficiency and blood stasis (KDBS) in terms of proteomics, lay a molecular biological basis for the differentiation of various blood stasis syndromes of EMT, establish a EMT microscopic syndrome differentiation and diagnosis system in terms of proteomics, discover the evolution principles and therapeutic targets of these EMT syndromes, and search their signifificant molecular markers and genetic intervention targets. METHODS: Six specimens from the ectopic and entopic endometrium tissues of patients with EMT in each syndrome, BS, QSBS as well as KDBS, in the early proliferative phase of the menstrual cycle, and 6 specimens from normal endometrium tissues in the early proliferative phase of the menstrual cycle were obtained. Three groups were formed in each syndrome by mixing two random specimens in equal amount, and then their respective two-dimensional electrophoresis graphs were obtained after total protein extraction. Finally, the detected differences in protein expression were identifified through matrix-assisted laser desorption Ionization-time of flflight mass spectrometry (MALDI-TOF/MS) and protein database. RESULTS: The results of differential proteins expressed in each syndrome were shown as follows: BS syndrome had 2 differential proteins in entopic endometrium and 1 differential protein in ectopic endometrium; KDBS syndrome had 3 in entopic endometrium and 3 in ectopic endometrium; and QSBS syndrome had 3 in entopic endometrium and 4 in ectopic endometrium. It was found out that annexin was highly expressed in both entopic and ectopic endometrium of KDBS syndrome; and myosin light chain 3 was highly expressed in both entopic and ectopic endometrium of QSBS syndrome. CONCLUSION: There are differential protein expressions among the 3 EMT syndromes, which might be the inner origin of syndrome characters, and these differential proteins might be the candidate biomarkers for the pathogenesis of various EMT syndromes.

Shedding new light on female fertility: The role of vitamin D.

In the last decades several studies suggested that vitamin D is involved in the modulation of the reproductive process in women due to the expression of VDR and 1α-hydroxylase in reproductive tissues such as ovary, uterus, placenta, pituitary and hypothalamus. Vitamin D has also a role in the regulation of sex hormone steroidogenesis. Increasing evidence suggests that vitamin D might have a regulatory role in polycystic ovary syndrome (PCOS)-associated symptoms, including ovulatory dysfunction, insulin resistance and hyperandrogenism. Vitamin D deficiency also has been reported to contribute to the pathogenesis of endometriosis due to its immunomodulatory and anti-inflammatory properties. Although most of the studies supported a role of vitamin D in the onset of these diseases, randomized controlled trials to assess the efficacy of vitamin D supplementation have never been performed. In this review we critically discuss the role of vitamin D in female fertility, starting from in vitro and in vivo studies, focusing our attention on the two most frequent causes of female infertility: PCOS and endometriosis.

Myeloperoxidase as a Potential Target in Women With Endometriosis Undergoing IVF.

As infertility is intimately associated with endometriosis, the levels of myeloperoxidase (MPO), a leukocyte enzyme and an oxidative stress marker, were determined in a case-control prospective study of 68 women with and without endometriosis undergoing in vitro fertilization in the outpatient fertility center within a tertiary care academic medical center. Measured values included plasma and follicular fluid (FF) concentrations of MPO, plasma estradiol, as well as oocyte quality, fertilization, implantation, and pregnancy rates in these women. In FF (mean ± standard error of mean [SEM]), the MPO concentrations (ng/mL) for controls were 4.3 ± 0.37, mild endometriosis (stages I-II) 3.9 ± 0.17, and moderate/severe endometriosis (stages III-IV) 16.6 ± 12.5 ( P < 0.0143). In FF, among patients supplemented with vitamins E and C, the MPO levels decreased significantly only in moderate/severe endometriosis from 25.3 ± 22.0 ng/mL to 4.9 ± 1.61 ng/mL, respectively. Plasma levels of MPO between groups did not change. Outcome data revealed a trend toward decreased percentage of mature oocytes, implantation rate, and clinical pregnancy rate with severity of endometriosis and MPO levels. Myeloperoxidase may be a potential oxidative stress target for endometriosis-associated infertility.

[Clinical effect evaluation of acupuncture combined with medication for prevention of endometriosis recurrence after surgery].

OBJECTIVE: To evaluate the effect and safety of acupuncture combined with Jiawei Mojie tablet for the prevention of endometriosis recurrence after surgery. METHODS: One hundred and six patients after the conservative surgery against endometriosis were randomly divided into a western medication group and an acupuncture and medication group(a combination group), 53 cases in each one. In the western medication group, gestrinone was applied orally. In the combination group, acupuncture and Jiawei Mojie tablet with oral administration were adopted. Xuehai (SP 10), Sanyinjiao (SP 6) and Guanyuan (CV 4) were mainly used, and acupoints based on syndrome differentiation were combined. After the surgery and when the menstruation was end, 3 to 5 days later the preventive treatment was used successively for 6 months. Patients were followed up after treatment and in 3 months, 6 months, and 18 months (two years after surgery) after treatment. The recurrence [Abdominal/perineal ultrasonic examination, serum cancer antigen 125 (CA125) test], menstruation score, endometriosis health profile-5 (EHP-5) and the safety index of serum alanine aminotransferase (ALT) were compared between the two groups. RESULTS: Eight patients (five cases in the western medication group and three cases in the combination group) were rejected because of dropping, and six ones (two cases in the western medication group and four cases in the combination group) discontinued. When patients were followed up in 18 months after treatment, the suspected recurrence rate through ultrasonic examination was 2.0% (1/50), and the unusual rate of serum CA125 text was 4.0% (2/50) in the combination group, which were apparently lower than 18.8% (9/48) and 25.0% (12/48) in the western medication group (both P < 0.01). After treatment and 3-month, 6-month, and 18-month following up, the menstruation scores of the combination group were obviously lower than those of the western medication group (P < 0.01, P < 0.05). After 18 months, the positive rate of EHP-5 in the combination group was 0 (0/50), which was markedly lower than 18.8% (9/48) in the western medication group (P < 0.01). After treatment and 3-month, 6-month, and 18-month following up, the unusual rates of serum ALT text in the combination group were lower than those in the western medication group (P < 0.01, P < 0.05). After treatment, the unusual rate of serum ALT text was not significantly different in the combination group compared with that before treatment (P > 0.05), and after treatment and 3-month, 6-month following up, the unusual rates of serum ALT text in the western medication group were remarkablely higher than those before treatment (P < 0.01, P < 0.05). After 18-month following up, the unusual rate of serum ALT text in the western medication group was not statistically significant compared with that before treatment (P > 0.05). CONCLUSION: Acupuncture combined with Jiawei Mojie tablet can effectively prevent endometriosis recurrence after surgery, and improve menstruation condition and life quality, which are less injurious for liver than gestrinone.

Effects of 1,25-Dihydroxy Vitamin D3 on Endometriosis.

CONTEXT: Endometriosis is an estrogen-dependent, chronic inflammatory disease. Recent studies have shown that vitamin D (VD) is an effective modulator of the immune system and plays an important role in controlling many inflammatory diseases. OBJECTIVE: The objective of the study was to clarify the in vitro effects of 1,25-dihydroxy vitamin D3 (1,25[OH]2D3) on human endometriotic stromal cells (ESCs) and to determine the serum levels of VD in endometriosis patients. DESIGN, PATIENTS, AND MAIN OUTCOME MEASURES: ESCs were isolated from ovarian endometrioma and cultured with 1,25(OH)2D3. Gene expression of IL-8, cyclooxygenase-2, microsomal prostaglandin E synthase-1, microsomal prostaglandin E synthase-2, cytosolic prostaglandin E synthase, 15-hydroxyprostaglandin dehydrogenase, matrix metalloproteinase (MMP)-2, and MMP-9 was examined using quantitative RT-PCR. The production of IL-8 and prostaglandin E2 was measured using an ELISA and an enzyme immunoassay. Viable cell number was assessed using a cell-counting assay, and DNA synthesis was assessed using the bromodeoxyuridine incorporation assay. Apoptosis was assessed using flow cytometry. The expression of inhibitory-κBα protein was detected using Western blotting. The serum levels of 25-hydroxyvitamin D3 and 1,25(OH)2D3 were measured by a RIA. RESULTS: In vitro studies showed that 1,25(OH)2D3 significantly reduced IL-1ß- or TNF-α-induced inflammatory responses, such as IL-8 expression and prostaglandin activity. 1,25(OH)2D3 also reduced viable ESC numbers and DNA synthesis but did not affect apoptosis. MMP-2 and MMP-9 expressions were reduced by 1,25(OH)2D3. 1,25(OH)2D3 inhibited nuclear factor-κB activation. The serum 25-hydroxyvitamin D3 levels were significantly lower in women with severe endometriosis than in the controls and women with mild endometriosis. Serum 1,25(OH)2D3 levels were not different between groups. CONCLUSIONS: VD modulates inflammation and proliferation in endometriotic cells, and a lower VD status is associated with endometriosis. Taken together, VD supplementation could be a novel therapeutic strategy for managing endometriosis.

The role of vitamin D in reproductive health--a Trojan Horse or the Golden Fleece?

In the last decade, vitamin D was in the spotlight in many fields of research. Despite numerous publications, its influence on reproductive health remains ambiguous. This paper presents an up-to-date review of current knowledge concerning the role of cholecalciferol in human reproduction. It covers various infertility issues, such as polycystic ovary syndrome, endometriosis, myoma-induced infertility, male infertility, premature ovary failure and in vitro fertilization techniques. Vitamin D deficiency, defined as serum concentration of 25-hydroxycalciferol of less than 50 nmol/L, is commonly noted more frequently than only in fertility clinic patients. It is a global trend that is observed in all age groups. The results of original publications dated up to 2015 have been summarized and discussed in a critical manner. Most experts agree that vitamin D supplementation is a necessity, particularly in women suffering from obesity, insulin resistance or small ovarian reserve, as well as in men with oligo- and asthenozoospermia if serum concentration should fall below 50 nmol/L (normal range up to 125 nmol/L). High concentration of vitamin D and its metabolites in decidua during the 1st trimester suggests its important role in the implantation process and a local immunological embryo-protection. On the other hand, evidence-based research did not prove a significant difference so far in ovulation stimulation or embryo development depending on vitamin D level. In one of the publications, it was also found that vitamin D binding protein (VDBP) has a molecular similarity to anti-sperm antibodies, and another one concluded that both low (<50 nmol/L) and high (>125 nmol/L) concentration of vitamin D are associated with decreased number and quality of spermatozoa in semen. Vitamin D is definitely not a Trojan Horse in reproductive health, since there were no adverse effects reported for vitamin D intake of up to 10,000 IU/day, but to proclaim it the Golden Fleece, more evidence is needed.

Effect of Bushenwenyanghuayu decoction on nerve growth factor and bradykinin/bradykinin B1 receptor in a endometriosis dysmenorrhea mouse model.

OBJECTIVE: To observe the effects of Bushenwenyanghuayu decoction (BD), a Traditional Chinese Medicine (TCM), on the serum concentration of nerve growth factor (NGF) and bradykinin (BK), and protein and mRNA levels of NGF and bradykinin B1 receptor (BKB1R) in a mouse model of endometriosis dysmenorrhea. METHODS: Seventy-five experimental female BALB/c mice were randomly divided into five groups, 15 mice each: sham, model, BD high dose (61.67 g/kg), BD low dose (15.42 g/kg), and gestrinone (0.4 mg/kg) groups. All the mice except for those in the sham group underwent auto-transplantation surgery and were gavaged estradiol valerate (0.5 mg/kg, daily for 12 days) after surgery. On the 12th day, 1 h after administration, writhing response was induced by intraperitoneal injection of oxytocin at 2 U/mouse. The writhing frequency and latency were recorded and the volume of the ectopic foci was measured. The concentration of serum NGF and BK was detected by enzyme-linked immunosorbent assay, the protein expression of NGF and BKB1R was tested by immunohistochemistry and western blotting, and NGF and BKB1R mRNAs were detected by real-time PCR. RESULTS: Compared with the model group, the volume of the ectopic foci in the treatment groups was significantly lower (P < 0.01), the writhing frequency was decreased (P < 0.05), and the writhing latency was prolonged (P < 0.01). Compared with the sham group, serum NGF and BK levels in the model group were significantly increased (P < 0.01). There were positive correlations for writhing frequency among the NGF and BK groups (P < 0.01). The serum NGF and BK levels were significantly lower in the treatment groups than the model group (P < 0.05). The protein expression of NGF, BKB1R was significantly decreased in the treatment groups compared with the model group (P < 0.01). NGF and BKB1R mRNA expression was significantly decreased in the treatment groups compared with the model group (P < 0.01). CONCLUSION: NGF and BK/BKB1R may play an important role in the development of endometriosis-associated dysmenorrhea, and BD was found to inhibit the development of endometriosis and relieve dysmenorrhea by influencing NGF and BK/ BKB1R mRNA and protein levels.

Serum Polyunsaturated Fatty Acids and Endometriosis.

Polyunsaturated fatty acids (PUFAs) are fatty acids containing 2 or more double bonds, and they are classified by the location of the last double bond. Omega 3 (n-3) and omega 6 (n-6) PUFAs are obtained through food sources including fatty fish and seed/vegetable oils, respectively, and they are important to a number of physiologic processes including inflammation. Previous work demonstrates suppressive effects of n-3 PUFAs on endometriotic lesions in animal models and decreased risk of endometriosis among women with high n-3 PUFA intake. Thus, we sought to determine the relationship between circulating levels of PUFAs and endometriosis in women. To do this, we performed a cross-sectional study of serum PUFAs and clinical data from 205 women undergoing in vitro fertilization (IVF). Serum PUFAs were measured using liquid chromatography coupled to tandem mass spectroscopy and included n-3 PUFAs such as α-linolenic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid and n-6 PUFAs such as linoleic acid and arachidonic acid. Multivariable logistic regression was used to determine relationships between specific and total serum PUFAs and patient history of endometriosis. Women with high serum EPA levels were 82% less likely to have endometriosis compared to women with low EPA levels (odds ratio = 0.18, 95% confidence interval 0.04-0.78).

[Effect of compound ezhu powder on serum levels of CA125 and CA19-9, and the expression of cyclin D protein in endometriosis patients].

OBJECTIVE: To study the effect of Compound Ezhu Powder (CEP) on serum levels of CA125 and CA19-9, and the expression of cyclin D protein in endometriosis patients, thus providing theoretical evidence for clinical application of CEP. METHODS: Totally 69 all endometriosis patients underwent surgical treatment at Department of Gynecology and Obstetrics, Tianjin Nankai Hospital from January 2011 to January 2013 were randomly assigned to group A (35 cases) and group B (34 cases). Meanwhile, 30 patients with uterine fibroids who prepared for surgical treatment during the same period were recruited as the control group. Patients in group A took EZP 3 months before surgery. No treatment was given to patients in group B and the control group. The serum CA125 level and the expression of cyclin D in the ectopic endometrium and the eutopic endometrium were detected in the 3 groups before surgery. RESULTS: The expression of cyclin D was higher in group A and group B than in the control group (P < 0.05). The serum levels of CA125 and CA19-9 were significantly lower in group A than in group B (P < 0.05). The expression of cyclin D in the ectopic endometrium was lower in group A than in group B, but with no statistical difference (P > 0.05). The expression of cyclin D in the eutopic endometrium was significantly lower in group A than in group B with statistical difference (P < 0.05). Meanwhile, the serum CA125 level was positively correlated with the serum CA19-9 level (r = 0.45, P < 0.05). CONCLUSIONS: The expression of cyclin D obviously increased in endometriosis patients, which was associated with the occurrence of endometriosis. CEP could lower serum levels of CA125 and CA19-9, and down-regulate the expression level of cyclin D, indicating its roles in inhibiting the cell cycle.

Resveratrol successfully treats experimental endometriosis through modulation of oxidative stress and lipid peroxidation.

BACKGROUND AND AIMS: The purpose of this study was to investigate the potential therapeutic efficiency of resveratrol in the treatment of experimental endometriosis in rats. SETTINGS AND DESIGN: Experimental study was carried out in a University hospital. MATERIALS AND METHODS: Endometriosis was surgically induced in 24 female rats. Four weeks after this procedure, the viability and dimensions of the endometriosis foci were recorded. Rats were then randomly divided into three groups: (1) Control group (n = 8); (2) low dose (10 mg/kg) resveratrol group (n = 8); (3) high dose (100 mg/kg) resveratrol group (n = 8). At the end of the 7-day treatment, blood samples were taken and laparotomy was performed. The endometrial implants were processed for biochemical, histological and immunohistochemical studies. STATISTICAL ANALYSIS USED: The Kruskal-Wallis H test and one-way ANOVA test were used. RESULTS: Resveratrol-treated rats showed significantly reduced endometriotic implant volumes (P = 0.004). After treatment, a significant and dose-dependent increase in activities of superoxide dismutase and glutathione peroxidase in serum and tissue of the rats in Group 2 and Group 3 was detected. Similarly, serum and tissue malonyl dialdehyde levels and tissue catalase levels were significantly higher in Group 3 than that of control animals. Histological scores and proliferating cell nuclear antigen expression levels were also significantly reduced in Group 2 and Group 3 than that of control group. CONCLUSION: In a rat endometriosis model, resveratrol showed potential ameliorative effects on endometriotic implants probably due to its potent antioxidative properties.