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Postmenopausal osteoporosis (PMOP) is a public health issue that needs to be addressed urgently. Patients are with decreased estrogen level, decreased estrogen receptor α (ERα) expression level, decreased osteoblast function, increased osteoclast activity, and imbalance of bone metabolic. ERα plays an important role in the occurrence and development of PMOP. In this paper, literature in the field of osteoporosis in recent ten years (2013-2023) are retrieved from PubMed, Web of Science, and CNKI. The role of ERα in bone mesenchymal stem cells, osteoblast progenitors, osteoblast progenitors, mature osteoblasts, osteoclasts, and osteocytes is summarized. The relevant anti-PMOP mechanism of ERα is expounded based on an example of icariin, a flavonoid component that significantly relieves PMOP. [ABSTRACT FROM AUTHOR]

Background and purpose: Aberrant N6-methyladenosine (m6A) modification caused by dysregulation of methyltransferase-like factor 14 (METTL14) plays an important role in the progression of various cancers, and it is unclear whether it is involved in the endometrial cancer (EC) progression. This study aimed to investigate the role of aberrant m6A modification caused by dysregulation of METTL14 in EC invasion and metastasis. Methods: Ninety-six EC patients who underwent curative surgery in Qinghai Provincial People’s Hospital from 2017 to 2021 were enrolled. RNA (70 pairs) or proteins (10 pairs) were isolated from frozen tissues for real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) or immunoblot analysis to assess METTL14 expression in EC. The expression of METTL14 and its correlation with clinicopathological features of EC were assessed. The biological effects of METTL14 in EC were determined in vitro and in vivo. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) combined with RNA sequencing (RNA-seq), and following m6A dot blot, MeRIP-RTFQ-PCR, RIP-RTFQ-PCR or dual luciferase reporter assays were employed to screen and validate the candidate targets of METTL14. Results: The mRNA expression and protein levels of METTL14 were significantly downregulated in EC compared with matched adjacent tissues. Compared with the METTL14 high expression group, the METTL14 low expression group had a significant increase in International Federation of Gynecology and Obstetrics (FIGO) stage, infiltration depth, lymphovascular invasion, lymph node metastasis and the number of cases of tumor metastasis (P

Background and purpose: Aberrant N6-methyladenosine (m6A) modification caused by dysregulation of methyltransferase-like factor 14 (METTL14) plays an important role in the progression of various cancers, and it is unclear whether it is involved in the endometrial cancer (EC) progression. This study aimed to investigate the role of aberrant m6A modification caused by dysregulation of METTL14 in EC invasion and metastasis. Methods: Ninety-six EC patients who underwent curative surgery in Qinghai Provincial People's Hospital from 2017 to 2021 were enrolled. RNA (70 pairs) or proteins (10 pairs) were isolated from frozen tissues for real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) or immunoblot analysis to assess METTL14 expression in EC. The expression of METTL14 and its correlation with clinicopathological features of EC were assessed. The biological effects of METTL14 in EC were determined in vitro and in vivo. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) combined with RNA sequencing (RNA-seq), and following m6A dot blot, MeRIP-RTFQ-PCR, RIP-RTFQ-PCR or dual luciferase reporter assays were employed to screen and validate the candidate targets of METTL14. Results: The mRNA expression and protein levels of METTL14 were significantly downregulated in EC compared with matched adjacent tissues. Compared with the METTL14 high expression group, the METTL14 low expression group had a significant increase in International Federation of Gynecology and Obstetrics (FIGO) stage, infiltration depth, lymphovascular invasion, lymph node metastasis and the number of cases of tumor metastasis (P＜0.05). Functionally, METTL14 inhibited the proliferation and invasive capacity of EC cells in vitro and in vivo. Mechanistically, METTL14-mediated demethylation of m6A resulted in post-transcriptional repression of estrogen receptor alpha (ERα). Furthermore, compared with METTL14, ERα induced oncogenic behavior of tumors. Conclusion: METTL14 attenuates ERα expression in EC cells in a m6A-dependent manner, thereby inhibiting tumor metastasis and invasion. [ABSTRACT FROM AUTHOR]

At present,hepatitis B cirrhosis still has a major impact on public health and national development. The development and progression of hepatitis B cirrhosis is associated with estrogen. Estrogen acts by binding to estrogen receptors,and the expression and functional status of estrogen receptors are affected by estrogen receptor gene polymorphisms. This article mainly introduces the structure and biological characteristics of estrogen and estrogen receptor α gene and their association with the pathogenesis of hepatitis B cirrhosis,and it is pointed out that the abnormal expression of estrogen may be associated with the development and progression of hepatitis B cirrhosis. However,due to the differences in genetic loci,environment,and location between studies,the results of the association of estrogen receptor α gene polymorphisms with hepatitis B cirrhosis may not be completely consistent. [ABSTRACT FROM AUTHOR]

In recent years, molecular signaling pathways related to bone metabolism has become a hot topic. Studies have revealed that signaling pathways associated with osteoporosis include Wnt/β-catenin, RANKL/RANK/OPG, NF-κB, PPAR-γ, PTH, MAPK, PI3K/Akt, Hedgehog and Notch signaling pathways. However, there has been no study concentrated on ERα-AMPK-Sirt1 signaling pathway. Estrogen receptor alpha (ERα), AMP-activated protein kinase (AMPK) and Sirtuin 1(Sirt1) can regulate the function of osteoblast or osteoclast by ligand binding, phosphorylation and deacetylation, respectively. Studies have indicated that ERα can directly increase the activity of LKB1, while LKB1 is the most important upstream protein kinase of AMPK, as a result, AMPK is activated. In addition, the mutual promotion of AMPK and Sirt1 can also modulate the autophagy or apoptosis of osteoblast; all these suggested that ERα-AMPK-Sirt1 signaling pathway may play an important role in the pathogenesis of osteoporosis. To sum up, it is necessary to start the research of ERα-AMPK-Sirt1 signaling pathway in order to better guide the development of new drugs and clinical practice. [ABSTRACT FROM AUTHOR]

BACKGROUND: Wnt signaling pathway plays an important role in formation, differentiation and mature of osteocytes. As negative regulators of Wnt signaling pathway, DKK1 and Sost play negative roles in regulating bone mass and osteoblast differentiation. XCT-790, an inhibitor of estrogen receptor-related receptor α, can regulate the effect of estrogen signaling pathway on osteoclast function. OBJECTIVE: To observe the effects of XCT-790 on the proliferation of MG63 cells transfected with DKK 1 and Sost overexpression adenovirus vector and the expression of low-density lipoprotein receptor-related protein (LRP) 5, bone morphologic protein 2, osteopontin, osteoprotegerin, and connective tissue growth factor (CTGF) protein. METHODS: MG63 cells were divided into eight groups: blank control, overexpressed DKK1 (ad-DKK1), overexpressed Sost (ad-Sost), overexpressed DKK1+Sost (ad-DKK1+Sost), XCT-790-intervened blank adenovirus, XCT-790-intervened ad-Sost, XCT-790-intervened ad-DKK1-Sost groups. The cell viability was detected by MTT assay. The alkaline phosphatase activity was detected by alkaline phosphatase kit. Ca2+ concentration was detected by flow cytometer. Expression of LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin and CTGF was detected by western blot assay. RESULTS AND CONCLUSION: Compared with the blank control group, ad-DKK1, ad-Sost, ad-DKK1+Sost and XCT-790-intervened blank adenovirus groups could significantly decrease the cell activity, alkaline phosphatase activity, increase calcium concentration and down-regulate the expression of LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin and CTGF (P < 0.05). In the XCT-790-intervented ad-DKK1, ad-Sost, ad-DKK1+Sost groups, the cell activity, alkaline phosphatase activity, and the expression of LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin and CTGF were significantly reduced, and the Ca2+ concentration was significantly increased (P < 0.05). In summary, XCT-790 can directly regulate Wnt signaling pathway, and can cooperate with DKK1 and Sost to inhibit Wnt signaling pathway, thereby decreasing the activity of osteoblasts, LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin, and CTGF protein expression, and further decreasing differentiation, proliferation and osteogenesis of osteoblasts. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the therapeutic effect of Leonuri Herba aqueous decoction on primary dysmenorrhea(PD) and explore the underlying mechanism in conjunction with untargeted metabolomics. Forty adult female rats were randomly divi-ded into a normal group, a model control group, ibuprofen(0.12 g·kg~(-1)) group, and high-and low-dose Leonuri Herba aqueous decoction(5 and 2.5 g·kg~(-1)) groups, with eight rats in each group. The PD rat model was prepared using intramuscular injection of estradiol benzoate combined with intraperitoneal injection of pitocin. Drugs were administered by gavage from the 4th day of modeling for 7 d. After the last administration, pitocin was injected intraperitoneally, and the writhing latency and writhing times within 30 min were recorded. The uterine and ovarian coefficients were determined. Estradiol(E&#95;2), progesterone(Prog), oxytocin(OT), cyclooxyge-nase 2(COX-2), prostaglandin E&#95;2(PGE&#95;2), prostaglandin F&#95;(2α)(PGF&#95;(2α)), and Ca~(2+) levels in uterine tissues were measured by ELISA and biochemical kits. Morphological changes in uterine and ovarian tissues were observed by hematoxylin-eosin(HE) staining. The protein expression of oxytocin receptor(OTR), prostaglandin E&#95;2 receptor 3(EP3), and estrogen receptor alpha(ERα) in uterine tissues was detected by immunohistochemistry. The mRNA expression of OTR, PGE&#95;2 receptors 1-4(EP1, EP2, EP3, and EP4), and PGF&#95;(2α) receptor(FP) in uterine tissues was detected by quantitative real-time PCR. Untargeted metabolomics analysis was performed by ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(LC-QTOF-MS) technology to screen potential biomarkers and enrich metabolic pathways. The results showed that Leonuri Herba was able to significantly reduce the writhing times in PD rats(P&lt;0.05 or P&lt;0.01), significantly reduce the uterine and ovarian coefficients(P&lt;0.01), and improve their histomorphology. After treatment with Leonuri Herba, PGE&#95;2 content was significantly increased(P&lt;0.05), COX-2, PGF&#95;(2α) and Ca~(2+) content, and PGF&#95;(2α)/PGE&#95;2 was significantly decreased(P&lt;0.05 or P&lt;0.01), and OT content was decreased, while E&#95;2 and Prog content tended to further increase in uterine tissues of PD rats. Correspondingly, OTR and EP3 protein expression was significantly downregulated(P&lt;0.05 or P&lt;0.01) and ERα protein expression was upregulated(P&lt;0.05) in uterine tissues. The mRNA expression of FP and EP4 in uterine tissues was significantly downregulated(P&lt;0.01), and the mRNA expression of EP1, EP3, and OTR showed a decreasing trend. The untargeted metabolomics results showed that 10 differential metabolites were restored in the plasma of PD rats after Leonuri Herba treatment. The results indicate that Leonuri Herba is effective in the prevention and treatment of PD, and the underlying mechanism may be attributed to the regulation of PGs synthesis and corresponding receptor binding.

Objective To further investigate the regulatory mechanism of estrogen on apolipoprotein M（ApoM）. Methods The mRNA and protein levels of ApoMwere assayed by reverse transcription polymerase chain reaction（RT-PCR） and Western blotting after incubation of bovine serum albumin conjugated estrogen（E2-BSA）at different concentrations in HepG2 cells after 24 h. ApoM mRNA were determined by RT-PCR after incubation of different estrogens with estrogen receptor α（ER-α）antagonist MPP in HepG2 cells after 24 h. Luciferase reporter assay and site-directed mutagenesis were used to analyze the regulatory region of ER-α. Electrophoretic mobility shift assay（EMSA）and chromatin immunoprecipitation（CHIP） were carried out to verify the interaction of ER-α with the regulatory region of ApoM. Results E2-BSA increased mRNA and protein levels of ApoM in the HepG2 cells with a dose-dependent manner，which could be abolished by addition of MPP. A regulatory region [from -1 580 to -1 575 bp（-GGTCA-）] upstream of the transcriptional start site of ApoM gene was essential for the basal transcriptional activity that regulated by the ER-α. Mutation in ER-α binding site [-1 580 to -1 575 bp（-GGTCA-）] of this region resulted in a decrease of luciferase activity，while cotransfection with ER-α expressing plasmids led to a remarkable increase of luciferase activity. EMSA and CHIP demonstrated that ER-α interacted with this region. Conclusion Estrogen can upregulate ApoM via ER-α. [ABSTRACT FROM AUTHOR]

Objective：To explore the association of single nucleotide polymorphism (SNP) of estrogen receptor α (ERα) gene with male infertility in Chinese Zhuang population. SNPs of PvuII and XbaI in ERα gene were checked as targets. Methods：A total of 96 infertile men were recruited as the observation group, and 76 fertile men as the control group. After routine semen analysis, the restricted enzyme digestion and DNA direct sequencing of PvuII and XbaI in ERα gene were performed. Results：The XbaIallele frequency and genotype distributions were significantly different between the control group and the observation group (all P<0.05). Genetic model analysis showed that the ERα gene XbaI were significantly different between the two groups under codominant (CC vs. TT) and recessive genetic model (both P<0.05). Subjects carrying the T allele frequency of XbaI had the decreased risk of infertility compared to the subjects carrying the G allele frequency. However, the SNPs of ERα gene Pvu II did not show the correlation with male infertility. Conclusions：We concluded that in this study ERα XbaI polymorphism was associated with male infertility. [ABSTRACT FROM AUTHOR]

Objective To analyze the regulation of estrogen receptor α (ERα) on truncated neurokinin-1 receptor (NK1R-Tr), and the influence of this regulation on cell proliferation in estrogen receptor-positive breast cancer cell lines. Methods The chromatin immune coprecipitation (CHIP) was used to observe the transcriptional regulation function of ERα on NK1R- Tr in breast cancer cells. Luciferase reporter gene assay was used to verify whether ERα played a positive regulatory role in the expression of NK1R-Tr. Western blot assay and real-time-PCR were used to detect the expression of ERα and NK1R-Tr in breast cancer cells, MCF-7 and T47D, as well as the expression of NK1R-Tr protein and mRNA level. NK1R-Tr levels were also detected after using estradiol (E2, ERα agonist) and small interfering RNA (knock out ERα). CCK- 8 and clone formation experimen were used to detect the proliferation ability of breast cancer cells after knocking out NK1R-Tr with small interfering RNAs. Results CHIP test and Luciferase reporter gene assay proved that ERα can positively regulate the expression of NK1R-Tr via the ERα sequences in the upstream of the NK1R-Tr gene promoter. The expression of NK1R-Tr at both protein level and mRNA level dropped in the estrogen receptor-positive breast cancer cell line MCF-7 upon knocking out ERα. After knocking out NK1R-Tr, the proliferation ability of estrogen receptorpositive breast cancer cells was lower than that of the control group. Conclusion The ERα positively regulates the expression of NK1R-Tr, resulting in the increased cell proliferation in estrogen positive breast cancer cells. [ABSTRACT FROM AUTHOR]

Objective To investigate the relationship between estrogen gene polymorphism and the incidence of melasma. Methods A total of 56 patients with chloasma blepharoplasty surgery (case group) and 39 healthy women with eyelid blepharoplasty (control group) were included in this study. The expression of estrogen receptor (ER) was detected by immunohistochemistry. The ERα gene PvuⅡ and XbaⅠrestriction site polymorphisms and RsaⅠ and AluⅠ restriction site polymorphism of ERβ gene were detected by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis in two groups. The relationship between the incidence of ER and the score of melasma was analysed. Results Results of immunohistochemistry showed that there were positive expressions of ERα and ERβ in dermal fibroblasts and endothelial cells of case group, while there was weak expressions of ERα and ERβ in control group. Spearman correlation analysis showed that there was a positive correlation between ERα, ERβ and melasma score (rs=0.462 and 0.512, P＜0.05). There was a significant difference in distribution of ERα gene XbaⅠ genotype between two groups (P＜0.05). With xx genotype reference, the value of OR exposed to X allele (Xx + XX) was 2.23 (95% CI: 1.41-3.89, P＜0.05). There was a significant difference in ERβ gene Alu Ⅰ genotypes between two groups (P＜0.05 ). With aa genotype reference, the value of OR exposed to the A allele (AA+Aa) was 1.58 (95% CI: 1.21-4.29, P＜0.05). There was a significant difference in Rsa Ⅰ genotype distribution between two groups (P＜0.05). With reference rr genotype, the value of OR exposed to R allele (RR + Rr) was 2.37 (95% CI: 1.19 - 6.33, P＜0.05). Conclusion The incidence of melasma is related with XbaⅠ ERα genotype, ERβ gene AluⅠ, RsaⅠ genotype polymorphisms. The gene mutations increase the risk of incidence of melasma. Persons with Xx, Aa and RR genotypes are susceptible to melasma. [ABSTRACT FROM AUTHOR]

Objective:To analyze the expression of SULT1E1，ERα, ERβ, PRA and PRB protein in uterine sarcoma in order to find their clinical significance. Methods:The immunohistochemical method was used to detect the expressions of SULT1E1，ERα, ERβ, PRA and PRB in 31 cases of endometrial stromal sarcoma，15 cases of leiomyosarcoma tissues，10 cases of uterine leiomyoma tissues and 10 cases of normal myometrium tissues far away the wrapping membrane of myoma. Results:①The positive rate of SULT1E1 protein in endometrial stromal sarcoma and leiomyosarcoma tissues is significantly lower than that in normal myometrium tissues (P<0.016 7).② The positive rate of ERα protein in endometrial stromal sarcoma and leiomyosarcoma tissues is significantly lower than that in uterine leiomyoma (P<0.0167). ③The positive rate of PRA protein in leiomyosarcoma is significantly lower than that in uterine leiomyoma (P<0.0167). ④ In endometrial stromal sarcoma tissues’ SULT1E1 had negative correlation with ERα，PRA and PRB (rs=-0.463, P=0.009; rs=-0.389, P=0.031;rs=-0.421, P=0.018, respectively)，and had no correlation with ERβ (rs=0.018, P=0.925).⑤ In endometrial stromal sarcoma tissues, ERβ had positive correlation with PRA and PRB (P<0.01). Conclusions：The diminution or deletion of SULT1E1 protein expression relates to the development uterine sarcoma；SULT1E1 has significantly negative correlation with ERα, PRA and PRB in endometrial stromal sarcoma, which may influence the development of endometrial stromal sarcoma. [ABSTRACT FROM AUTHOR]

The classical estrogen receptor (ER) and its variants, membrane estrogen receptor (mER) involved in mediating different signaling effects of estrogen. Studies have shown estrogen is closely related to the occurrence and development process of endometrial cancer, while excessive estrogen stimulation and lack of progesterone antagonist is widely accepted as the main cause of endometrial cancer and the function of estrogen is mediated by estrogen receptor. Learning the possible mechanisms of the occurrence and development of endometrial cancer and finding new ways for its diagnosis and treatment through estrogen receptor also showing great significance. This paper reviews the structure and function of estrogen receptor, and analyses its relationship with endometrial cancer. [ABSTRACT FROM AUTHOR]

Objective To investigate the genetic polymorphisms in the estrogen receptor (ER) alpha gene at position 29 (ERα-29) in patients with chronic hepatitis B (CHB) from Han nationality and Hui minority populations in Gansu Province, China. Methods Polymorphisms of ERα-29 were analyzed in 84 Han and 84 Hui CHB patients who were admitted to our hospital from August 2012 to November 2013 by polymerase chain reaction-restriction fragment length polymorphism analysis. Allele frequencies were determined by gene counting and tested for expectations of Hardy-Weinberg equilibrium. Unconditional logistic regression analysis was applied to adjust the confounding factors. Comparison of the genotype and allele frequencies was performed by chi-square test. ResultsThe genotype frequencies of ERα-29 in Han versus Hui CHB patients from Gansu Province were as follows: TT, 52.4% vs 60.7%; TC, 40.5% vs 34.5%; CC, 7.1% vs 4.8%. The allele frequencies were 72.6% vs 78.0% for the T allele and 27.4% vs 22.0% for the C allele. No significant differences in the allele and genotype frequencies of ERα-29 were found between Han and Hui CHB patients in Gansu Province (P＞0.05). Conclusion The genetic polymorphisms in ERα-29 are not significantly different between Han and Hui CHB patients in Gansu Province, China. [ABSTRACT FROM AUTHOR]

This study aims to explore the anti-inflammatory and hemostatic effects of the total extract of Clinopodium chinense(TEC), total saponins of C. chinense(TSC), and total flavonoids of C. chinense(TFC) in female rats with abnormal uterine bleeding(AUB), and the possible mechanism. Mifepristone(i.g., 12.4 mg·kg~(-1)) and misoprostol(i.g., 130 μg·kg~(-1)) were used to induce AUB in SD female rats conceiving on the same day. Then the AUB rats were randomized into model group, TEC group, TSC group, TFC group, Yimucao Granules(LG) group, and estradiol valerate(EV) group, with 8 rats in each group. Another 8 non-pregnant female rats were selected as normal group. During the experiment, each group was given the corresponding drug by gavage once a day for 7 days. After the administration, blood and uterine tissue were collected. The uterine bleeding volume was measured by ultraviolet spectrophotometry and the pathological changes of endometrium were observed based on hematoxylin-eosin(HE) staining. In addition, the microvessel density of endometrium was determined by immunohistochemistry, and the content of thromboxane B2(TXB2), 6-keto-PGF&#95;(1α), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in plasma and levels of lutenizing hormone(LH), follicle stimulating hormone(FSH), estradiol(E&#95;2), and progesterone in serum were detected by enzyme-linked immunosorbent assay(ELISA). The mRNA and protein expression of estrogenreceptor α(ERα), progesterone receptor(PR), matrix metalloproteinase(MMP)-2, MMP-9, and vascular endothelial growth factor(VEGF) in uterine tissue was determined by Western blot. Compared with the model group, TEC, TSC, and TFC can reduce uterine bleeding volume, alleviate the pathological damage of endometrium, and increase the microvessel density in endometrium. Moreover, TEC and TSC can significantly raise plasma TXB2 level and ratio of TXB2 to 6-keto-PGF&#95;(1α), and TEC and TFC can significantly reduce the levels of IL-6 and TNF-α. In addition, TEC significantly elevated serum progesterone level and TFC significantly increased serum levels of E&#95;2, FSH, and LH. TSC can significantly raise serum progesterone and FSH levels. In addition, TEC can significantly down-regulate the protein expression of PR, MMP-2, and VEGF and TSC significantly reduced the expression of MMP-9. TFC significantly decreased the expression of PR, MMP-9, and VEGF, and up-regulated the expression of ERα. In conclusion, TEC, TSC, and TFC all show therapeutic effects on AUB, particularly TEC. TSC exerts the effects by enhancing the coagulation function and promoting endometrial repair, and TFC by regulating estrogen levels and reducing inflammatory response. This study reveals the mechanism of C. chinense against AUB and also explains the holistic characteristics of Chinese medicine.

Objective: To compare the effect of electroacupuncture (EA) on "Guanyuan" (CV4) or sensitization points in mice with polycystic ovary syndrome (PCOS), so as to explore its mechanisms underlying improvement of PCOS., Methods: In the first part of this study, 26 female ICR mice were randomized into control group (8 mice) and model group (18 mice). The PCOS model was established by gavage of bisphenol A (BPA) at a dose of 100 mg/kg, and the control group were gavage of equal volume of corn oil, once daily, 5 days a week for 4 conse-cutive weeks. Evans blue (EB) dye (0.1 mL/10 g) was injected into the caudal vein after modeling. The size, number and distribution of EB exudation points at the skin were observed. In the second part of this study, 32 mice were randomized into control, model, EA-CV4 and EA-sensitization points groups (8 mice in each group). EA (2 Hz, 2 mA) was applied to the sensitization points or CV4 for 20 min, once daily, 5 days a week for 4 conse-cutive weeks. The body weight was measured once a week for 8 consecutive weeks. The behavior changes were evaluated by open field test and elevated plus maze test. H.E. staining was used to observe the histopathologic changes of the ovary tissue. Serum level of estradiol (E 2 ) was measured by ELISA. The expressions of estrogen receptor α (ER-α) in ovarian and uterine tissues were detected by Western blot and immunofluorescence., Results: (1) In PCOS mice, the EB exudation points were found to overlap the lower abdomen, lumbosacral, chest, back and lower limbs regions, and the number of EB points was significantly more than that of the control group ( P <0.01). (2) After the intervention and compared with the control group, the ovaries showed polycystic changes and an increase of atresia follicles with a larger diameter, the activity time in the central area, the total distance of movement, the times of open-arm entries, the duration in open-arm, the serum E2 content and the expression of ER-α in ovarian tissue were significantly decreased ( P <0.01, P <0.05), while the mice's body weight and the expression of ER-α in uterine tissue were increased ( P <0.05) in the model group. After the intervention and compared with the model group, a small number of normal follicles and corpus luteum were observed under microscope, the activity time in the central area, the total distance of movement, the times of open-arm entries, the duration in open-arm, the serum E2 content and the expression of ER-α in ovarian tissue were significantly increased ( P <0.05, P <0.01), while the mice's body weight and the expression of ER-α in uterine tissue was decreased ( P <0.05) in both EA-CV4 and EA-sensitization points groups., Conclusion: EA at sensitization points and CV4 can regulate the expression of estrogen and ER-α in PCOS mice, and improve the anxiety like behavior. EB exudation points on the body surface can not only reflect the functional changes of organs, but also treat diseases through body surface stimulation, suggesting a dual role in diagnosis and treatment.

Objective: To detect the expression level of small ubiquitin-like modifier protein-1 (SUMO-1) and estrogen receptor alpha (ERα), and their correlation in endometrial carcinoma. Methods: 50 cases of endometrial carcinoma samples and 35 cases of normal endometrial samples were collected in our hospital from 2006 January to 2013 December. The expression of SUMO-1 and ERα were detected in all tissues by immunohistochemical method, and their co-localization expression was detected by immunofluorescence method. Then the relationship between SUMO-1 and ERα, and their express level and the clinicopathological parameters were analyzed respectively. Results: The positive rates of SUMO-1 protein in endometrial carcinoma was higher than that in control group, while the expression of ERα protein was the opposite, and the difference between the two groups was statistically significant (P<0.05). The positive rate of SUMO-1 and ERαin endometrial carcinoma patients were differ in different ages, menopausal status and different histological grades (all P<0.05), but the difference was not significant in different pathology subtypes (P>0.05). The expression of SUMO-1 and ERα protein had good consistency (Kappa=0.875, Z=5.473, P=0.001). Conclusions: SUMO-1 and ERα were co-localization in endometrial carcinoma, and may play a synergistic role in the process and development of endometrial carcinoma. [ABSTRACT FROM AUTHOR]

Objective: To observe the effects of estrogen and its receptor modulators on the expressions of NF-κB and MMP-9 mRNAs in the thoracic aorta tissues of the castrated APOE-/- mice. Methods: The model of atherosclerotic plaque was developed by 44 castrated APOE-/- mice. These mice were randomly divided into 6 groups: ➀ the control group: saline 0.2 mL/d; ➁ the HT group - progynova 0.13 mL/d plus dydro-gesterone 0.13 mL/ d; ➂ the ICI+E2 group-ICI subcutaneous injection 0.13 mL/week plus progynova 0.13 mL/d; ➃ the ICI+HT group: ICI subcutaneous injection 0.13 mL/week, progynova 0.13 mL/d plus dydro-gesterone 0.13 mL/d; ➄ the PHTTP+E2 group: progynova 0.13 mL/d plus PHTTP 0.13 mL/d; 6 the PHTTP+HT group - progynova 0.13 mL/d plus dydro-gesterone 0.13 mL/d plus PHTTP 0.13 mL/d. After 8 weeks treatment, total RNA were extracted from the atheroselerotic plaque tissues. Expressions of NF-κB and MMP-9 mRNAs were detect using real-time PCR. Results: There was not significant difference in the expression of MMP-9 mRNA among 6 groups (P>0.05), while there was significant difference in NF-κB mRNA among 6 groups (P>0.05). Compared with the control group, the PHTTP+E2 group and the PHTTP+HT group had significantly higher expressions of NF-κB mRNAs (P=0.047 ... P =0.035). The NF-κB expression in the ICI+HT group was significantly lower than that in the PHTTP+HT group (P=0.028). However, there were not significantly differences in NF-κB expression among the control group, HT group, ICI+E2 group and ICI + HT group (compared with the control group, the values of P were 0.072,0.068 and 0.054, respectively). Conclusions: The hormone replacement therapy mediated by ERα (PHTTP+E2 and PHTTP+HT) increases the expression of NF-κB in atherosclerotic tissues, which indicated that ERa might increase the instability of atherosclerotic. [ABSTRACT FROM AUTHOR]

Reduced bioavailability of estrogen increases osteoporosis and fracture risk in postmenopausal women significantly, but the mechanisms by which estrogen regulates bone mass are incompletely understood. In recent years, mouse models with cell-specific deletion of the estrogen receptor α (ERα) have provided novel insights into the function and signaling of ERα which provide opportunity to develop new drugs for preventing and treating osteoporosis. Illuminating the role of ERα in different bone cells, including chondrocyte, osteoclast, osteoblast progenitor cells, osteoblast and osteocyte to figure out the mechanisms of ERα in the pathogenesis of osteoporosis is of great importance to find new targets for osteoporosis. [ABSTRACT FROM AUTHOR]

The coordination and unification of Yin and Yang are the basis of normal human life activities. Along with the age growth and aging of the body, women will suffer from menopausal syndrome during menopause. In addition to the significant changes in the genital system, there are also pathological manifestations in estrogen target points including bone, nerve and cardiovascular systems, due to the imbalance of Yin and Yang. Besides the insufficiency of estrogen, the main cause of menopausal syndrome is the changes in the response of target organs to estrogen. In other words, the biological effects mediated by estrogen receptor(ER) alpha and beta subtypes in target cells are often different or even opposite; the changes of expression level and ratio of ERα and ERβ are also important causes for the abnormal estrogenic effects in target organs and the imbalance of Yin and Yang of the body. Therefore, on one hand, the therapeutic mechanism of drugs is ER-mediated estrogenic effect. On the other hand, the drugs have a regulatory effect on ER subtype expression in target cells and Yin-Yang state in target organs and even organisms, so as to cause further changes in the response of target cells to estrogen or estrogenic components, and exert its therapeutic effects. This paper reviews the pharmacological mechanism of gynecological traditional Chinese medicine in harmonizing Yin and Yang in estrogen-positive target cells and the clinical efficacy in the following aspects, including estrogen and its mechanism, the estrogenic effect of ER in traditional Chinese medicine and the mechanism of ER subtype in balancing Yin and Yang and mediating and regulating the main target tissues in menopausal syndrome treatment.

Objective: To elucidate whether metformin could regulate the mRNA expression level of estrogen synthetase and ER in human uterine leiomyoma tissues. Methods: (1) Seventeen pairs of uterine leiomyoma tissues and adjacent myometrium (>2 cm) were collected from patients underwent hysterectomy in Peking University First Hospital between December 2016 and January 2017. Real-time PCR was used to measure the mRNA expression level of estrogen synthetase [including cytochrome P450 cholesterol side chain cleavage enzyme (P450scc), cytochrome P450 17α-hydroxylase (P450c17), 3-beta-hydroxysteroid dehydrogenase type 2 (3β-HSD-2), 17-beta-hydroxysteroid dehydrogenase type 1 (17β-HSD-1) and aromatase cytochrome P450 (P450arom)] and ER (including ERα and ERβ) in the uterine leiomyoma tissues and adjacent myometrium. (2) Uterine leiomyoma cells derived from uterine leiomyoma tissues were identified by immunocytochemistry method and cultured to the third generation. The treatment groups were cultured with different concentrations of metformin (10, 50 and 100 μmol/L) for 48 hours, and the control group was cultured with deionized water for 48 hours. The mRNA expression level of estrogen synthetase and estrogen receptor subtypes were measured by real-time PCR. Results: (1) P450scc, P450c17, 3β-HSD-2, 17β-HSD-1, P450arom mRNA median expression levels were 112, 4, 13, 42 and 194 in the uterine leiomyoma tissues, and were respectively 114, 5, 11, 32 and 6 in the myometrium. Compared to those of the myometrium, 3β-HSD-2 and P450arom mRNA expression levels in the uterine leiomyoma tissue were significantly higher ( P< 0.05), while there were no significant change of mRNA expression levels among P450scc, P450c17 and 17β-HSD-1 ( P> 0.05). ERα and ERβ mRNA median expression levels were 208 and 116 in the uterine leiomyoma tissues, and were 24 and 95 in the myometrium. Compared to that of the myometrium, ERα mRNA level in the uterine leiomyoma tissue was significantly higher ( P= 0.001), while there were no significant change of ERβ mRNA level ( P= 0.193). (2) After cultured with different concentrations of metformin (10, 50 and 100 μmol/L), the P450arom mRNA levels in the uterine leiomyoma tissues were 9±4, 8±5 and 8±3 respectively in the treatment groups and was 16±5 in the control group. Compared to that of the control group, P450arom mRNA expression levels in the treatment groups were significantly declined ( P< 0.05). There were no significant different change of mRNA expression levels among 3β-HSD-2, ERα and ERβ between the treatment groups and the control group ( P> 0.05). Conclusions: Metformin could down-regulate the mRNA expression level of aromatase in the uterine leiomyoma cells. These results indicate that metformin may inhibit the local estrogen synthesis and therefore suppress the development of uterine leiomyoma.

Objective: To observe the effects of moxibustion at "Guanyuan" (CV 4) and "Sanyinjiao" (SP 6) on bone morphology, metabolism and ERα of bone marrow mesenchymal stem cells (MSCs) in the ovariectomized rats and explore the underlying mechanism of moxibustion at Guanyuan (CV 4) and Sanyinjiao (SP 6) on the regulation of bone metabolism., Methods: A total of 60 SD rats were randomized into a normal group (20 rats) and an ovariectomy group (40 rats). In the normal group, no any treatment was provided. In the ovariectomy group, the classical ovariectomy was adopted to set up the osteoporosis model. In the 13th week after ovariectomy, 10 rats were collected from each of the normal group and the ovariectomy group for model identification. After model identification, the ovariectomy group was subdivided into a model group, an estradiol group and a moxibustion group, 10 rats in each one. In the normal group and the model group, the gavage was provided with 2 mL 0.9% sodium chloride solution, once a day. In the estradiol group, the gavage was provided with 17β-estradiol 100 μg/(kg•d), once a day. In the moxibustion group, moxibustion was applied at "Guanyuan" (CV 4) and "Sanyinjiao" (SP 6), 15 min at each acupoint, once a day. The 6 treatments were as one course, with 1 day of interval, 12 courses were required. After 12-week intervention, the dual-energy X-ray was adopted to determine the bone mineral density (BMD) and bone mineral content of the whole body in living condition. After sacrificed, HE staining was adopted to observe femur structure, the enzyme-linked immunosorbent assay was to determine the absorbance of estradiol (E 2 ) and alkaline phosphatase (ALP) in serum and the real-time quantitative PCR method was to determine the mRNA expressions of ERα of MSCs in tibia and femur., Results: Compared with the model group, BMD was increased obviously in the pelvis and spine in the moxibustion group and the estradiol group ( P <0.05, P <0.01). Compared with the model group, the bone mineral content was higher in the rib, pelvis and spine in the moxibustion group and the estradiol group, there was no statistically significant difference (all P >0.05). Compared with the model group, the bone trabeculas were stronger and well connected in the moxibustion group, indicated by HE staining. Compared with the model group, the concentrations of E 2 in serum were increased obviously in the estradiol group and the moxibusiton group (both P <0.01), and ALP concentrations reduced obviously in the estradiol group and the moxibustion group (both P <0.001), the mRNA expressions of ERα in MSCs increased in the estradiol group and the moxibustion group (both P <0.001)., Conclusion: Moxibustion at "Guanyuan" (CV 4) and "Sanyinjiao" (SP 6) effectively increases BMD and bone strength in the ovariectomized rats and the mechanism may be related to the improvement of serum E 2 concentration, the decrease of serum ALP concentration and the up-regulation of mRNA expression of ERα in MSCs.

Objective: To determine the effects of arsenic and estrogen receptor antagonist (ICI182, 780) on the expression of estrogen receptor beta (ERβ) in alveolar Ⅱ epithelial cells (AECⅡ) of female and male mice., Methods: Nineteen or twenty day fetus mice were obtained through caesarean section of ICR mice. Purified AECⅡ cells were separated from the female and male fetus,respectively,and confirmed using immunofluorescence staining. The cells were exposed to sodium arsenite (NaAsO 2 ) at a low,medium,or high dosage determined by MTT and cultured for 24 h. The NaAsO 2 (5 μmol/L) exposed cells were compared with those treated (for 24 h) with dimethyl sulfoxide (DMSO) or ICI182, 780 (1×10 -4 mol/L). Apoptosis rates of the cells were measured by flow cytometry. Real-time fluorescence quantitative PCR method and Western blot technique were used to detect the expression of ERβ mRNA and protein in AECⅡ., Results: Purity of AECⅡ cells reached (87.0±2.5)%. NaAsO 2 exposure was set at a concentration of 0.5 (low),1.25 (medium),and 5 (high) μmol/L. The cells exposed to medium and high dosage of NaAsO 2 had higher apoptosis rates than the blank controls ( P <0.05),without sex differences. Female cells exposed to medium and high dosage of NaAsO 2 had higher levels of expressions of ERβ mRNA and protein than the blank controls ( P <0.05) and male cells exposed to the same dosage of NaAsO 2 ( P <0.05). No significant differences were found in the expressions of ERβ mRNA and protein between the exposed male cells and the blank controls. ICI182, 780 lowered the expression levels of ERβ mRNA and protein in the female exposed cells ( P <0.01)., Conclusion: Arsenic exposure increases expressions of AECⅡ's ERβ,more so in female cells than in male cells. This can be blocked by estrogen receptor antagonists., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).)

Objective: The aim of this study is to analyze the different expression and function of androgen receptors (AR) and estrogen receptor (ER-α) in laryngeal squamous cell carcinoma (LSCC). In combination with the expression of prolactin receptor (PRLR), we analyzed the prognostic impact of three receptors on the laryngeal carcinoma. Method: In this study, immunohistochemistry and reverse transcription polymerase chain reaction (RT-PC) analysis were performed on the tumor tissues and adjacent normal tissues in 112 LSCC patients (95 males and 17 females). We found that hormone receptor expression is closely related to the clinical tumor lesions and overall survival data. Result: The expression of AR, ER-α and PRLR in tumor tissues were much higher than those in adjacent normal tissues ( P >0.05) at both protein and mRNA levels. The higher PRLR level indicate poor survival in LSCC patients ( P =0.02), while higher ER-α expression could influence the survival with considerable trend toward significance ( P =0.06). Furthermore, the higher expression of ER-α in tumours were corresponding with PRLR cytoplasmic higher level expression ( r =0.802, P =0.04). This mutual promoted effect between ER-α and PRLR possibly suggests potential mechanisms among those sex related hormone receptors in laryngeal cancer. Conclusion: It has become increasingly credible that the sex related hormone receptors play an important role in the development of LSCC.

Objective: To investigate the effect of equol on the proliferation of colom cancer cells and to explore the mechanisms., Methods: Colon cancer cells (DLD1,HCT15,COLO205,LOVO,SW480) were incubated, the cell proliferation was identified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay. Reverse transcription PCR and Western blot were used to measure the mRNA and the protein expression of estrogen receptor and nuclear factor (erythroid-derived 2)-like 2 (Nrf2)in the colon cancer cells, respectively. Moreover, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay was used to investigate the effect of estrogen receptor(ER) inhibitor,ERα agonist, and estrogen receptor ERβagonist on the cell proliferation., Results: ERα was faintly expressed in the DLD-1 and HCT-15 cells. However, ERβ expression in DLD1, HCT15, COLO205, LOVO, and SW480 colon cancer cells. Different concentrations of equol (0, 0.5, 1, 5, 10 μmol/L) significantly inhibited the growth of HCT-15 cell with the expression of ERα and ERβ.More-over, different concentrations of equol (0, 0.5, 1, 5, 10 μmol/L) significantly inhibited the growth of LOVO, and SW480 cells with the ERβ expression in a dose-dependent manner as demonstrated with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay. mRNA expressions of ERα and ERβ in HCT-15 were stimulated significantly. Western blotting proved that the protein expressions of ERα and ERβ increased with the increasing of equol dose. Moreover we found significant difference of Nrf2 protein expression in HCT-15 cell stimulated by different concentrationss of equol. After the similation of estrogen receptor inhibitor, ERα agonist, or ERβ agonist, we found that only dif-ferent concentrations of ERβ agonist(0, 1, 10, 100, 1 000, 10 000 nmol/L) significantly inhibited the growth of HCT-15, LOVO, and SW480 in adose-dependent manner. Estrogen receptor inhibitor and ERα agonistdid not present significant effect on the cell proliferation of HCT-15, LOVO, and SW480., Conclusion: Equol inhibited the colon cancer cell proliferation by its estrogenic activities and antioxidant activities.

Purpose: To evaluate the possible role and mechanism of estrogen in the morphologic alteration process of mandibular condyle., Methods: Forty healthy non-pregnant female 6-week-old Sprague-Dawley rats were randomly divided into control group, contraceptive group (intragastric administration with contraceptive), contraceptive + normal saline (intragastric administration with contraceptive and normal saline) and contraceptive +estradiol group(intragastric administration with contraceptive and estradiol) equally. The serum levels of estradiol were detected respectively before the experiment, on the twelfth week and sixteenth week. The rats were sacrificed after the experiment and the expression of ERα, ERβ of condyle was detected and histologic observation of the condyle was performed. SPSS11.0 software package was used for statistical analysis., Results: The serum estradiol concentration of the other 3 groups were significantly lower than control group at 12 week (P<0.05)； the serum estradiol concentration of contraceptive + estradiol group was significantly higher than contraceptive + normal saline group on sixteenth week (P<0.05). H-E and immunohistochemistry showed that compared with control group, not only retrogression occurred in rat condylar cartilage but also the expression of ERα and ERβ of condylar cartilage was decreased(P<0.05) in contraceptive group; Compared with the contraceptive + normal saline group, the retrogression was repaired and ERαexpression was increased(P<0.05) while ERβ was not increased (P>0.05) in contraceptive + estradiol group., Conclusions: The lowered serum estrogen would lead to retrogression in condylar cartilage. Estradiol valerate could be used to raise concentration of serum estrogen, increase the expression of ERα and slow down the process of condylar cartilage retrogression through ER pathway.

Objective: To investigate the regulatory functions and molecular mechanisms of miR211 in hepatocellular carcinoma (HCC)., Methods: Real-time reverse transcription-PCR was used to analyze the expression of miR-211 in 20 paired clinical specimens of HCC and adjacent noncancerous tissues.QGY7703 and HepG2 cells with stimulation or inhibition of miR-211 expression were used to evaluate the effects on malignant phenotypes with the transwell invasion assay. Candidate target genes of miR-211 were identified by bioinformatic screening and verified by the EGFP report assay, real-time PCR and western blotting. Moreover, the regulatory functions of miR-211 on the target genes were investigated by RNA interference and cell phenotype assays., Results: miR-211 was up-regulated in HCC tissue specimens (t =6.26, P < 0.01).HCC cells overexpressing miR-211 showed greater invasive capacity than cells with inhibited expression (QGY-7703:t =12.59, P < 0.01; HepG2: t =17.82, P < 0.01). Estrogen receptor a (ESR1) was identified and validated as a target gene of miR-211; knockdown of ESR 1 promoted HCC invasive capacity (QGY-7703:t =8.97, P < 0.01; HepG2:t =29.31, P < 0.01)., Conclusion: miR-211 promotes invasion of carcinoma cells by directly targeting ESR1.

Estrogen receptors (ERs), including two sub-types ERα and ERβ, belong to the steroid hormone superfamily of nuclear receptors. ERα distributes in the male reproductive system and plays a crucial role in the regulation of male reproduction through estrogen-dependent and -independent ways. In this article, we mainly reviewed the molecular structure, mode of action and location of ERα in the male reproductive system, and explored the mechanism of ERα in regulating the male reproductive system by analyzing different animal models of disrupted ERα.

Objective: To study the expression of the estrogen receptor (ER) and type II collagen in the mandibular condylar cartilage of the human embryo., Methods: The expression and localization of ERalpha, ERbeta, and type II collagen in the mandibular condylar cartilage of the human embryo were examined through hematoxylin-eosin (HE) and immunohistochemistry staining., Results: Type II collagen was primarily localized in the transitional and hypertrophic layers of the condylar cartilage. ERa was mostly expressed in the transitional and hypertrophic cartilaginous layers of the condylar cartilage. ERa was evenly distributed in the cell, whereas ERbeta was localized in the nuclei. No expression of type II collagen and ER was found in the fibrage and the proliferative layer although minimal expression was found in the calcified cartilage., Conclusion: The distribution of ER and type II collagen in the mandibular condylar cartilage was consistent. Estrogen can selectively combine with different subtypes of ER that regulate the ability of the condylar cartilage cells to secrete type II collagens.

Purpose: To evaluate the effect of estradiol on the expressions of estrogen receptor in mandibular condylar cartilage cell (MCC)., Methods: The MCC were harvested from neonatal rats. The experimental groups was cultured in medium with different concentration(10(-3),10(-6),10(-9),l0(-12) mmol/L) of estradiol(E2), while the control group was cultured in medium with ethanol. The ERα mRNA and ERβ mRNA in cells were detected with RT-PCR.The data was processed with SPSS10.0 software package., Resluts: Both ERα and ERβ expression were observed in MCC. Exogenous 17β-E2 raised the gene expression of ERβ and lowered the expression of ERα. Under the concentration of 10-9 mmol/L(physiological condition), the up-regulation of ERα mRNA was found most obviously., Conclusions: E2 can raise the expression of ERα and decrease the expression of ERβ, and the effect of up grading is the most obvious under physiological concentration.

Objective: To determine whether polymorphisms in the estrogen receptor (ER) genes are associated with chronic periodontitis in a Chinese population overall the Shanxi territory., Methods: 109 patients with chronic periodontitis and 99 healthy controls were recmited in this study. Genomic DNA was extracted from oral mucosa swab sample of each subject by the Chelex-100 method. Determination of the ER-alpha and -beta polymorphisms was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), respectively., Results: The analysis of the Xba I polymorphisms in ER-alpha gene revealed significant differences between patients and controls. The detection frequency of XX genotype was significantly higher in the chronic periodontitis patients than in control subjects. The difference between the female chronic periodontitis patients and healthy controls was statistically significant, but no difference was found between the male patients and controls. Nevertheless, no significant association was noted in the frequency of both Rsa I and Alu I polymorphisms in ER-beta gene between chronic periodontitis and controls., Conclusion: It is indicated that the ER-alpha XX genotype may be a susceptible indicator for chronic periodontitis in female Han Chinese subjects.

Objective: The aim of this study was to demonstrate the temporal and spatial expression of estrogen receptor (ER), 17beta-hydroxysteroid dehydrogenase (17beta-HSD) in uterine endometria and the endometrial-myometrial interface (EMI) of adenomyosis and the effects of estrogen activity-related molecules on the occurrence of adenomyosis., Methods: Thirty-three cases of normal endometria (and myometrial) and eighteen cases of endometria (and myometrial) with adenomyosis were collected. Immunohistochemical assay was performed to locate the ERalpha, ERbeta, 17beta-HSDI and 17beta-HSDII in endometria and EMI., Results: The ERalpha positive cell number in glandular epithelial cells at the early proliferative phase increased evidently in adenomyosis (90%), while it was 60% in normal endometria. We found ERalpha signal in cytoplasm in glandular epithelial cells of adenomyosis endometria as well as in nucleus. Compared with normal endometrium (early proliferative phase: +; late proliferative phase: ++; late secretory phase: +), eutopic endometrium with adenomyosis exhibited a higher level of 17beta-HSDI (early proliferative phase: ++; late proliferative phase: +++; late secretory phase: ++). The intensity of 17beta-HSDIIin glandular epithelial cells of eutopic endometrium with adenomyosis (early proliferative phase: +++; secretory phase: ++++) was also higher than that of normal endometrium (early proliferative phase: - approximately ++; secretory phase: +++). Higher intensities of ERalpha, ERbeta and 17beta-HSDIand lower intensities of 17beta-HSDII were observed in EMI than in the eutopic endometrium of adenomyosis., Conclusions: The elevation of ERalpha positive cell number, 17beta-HSDI level as well as the insufficient compensation of 17beta-HSDII in eutopic endometrium with adenomyosis and the change in expression pattern of ERalpha, ERbeta, 17beta-HSDI and 17beta-HSDII in EMI lead to the local enhancement of estrogen effect, which would promote cell proliferation.

Objective: To investigate the possible mechanism by which estrogen regulates apoptosis through the estrogen receptor., Methods: By means of fluorescence immunocytochemistry, the present study investigated the distribution of Bcl-2 and the colocolization of Bcl-2 and ERalpha immunoreactivity in the hippocampus of 10 Alzheimer's disease (AD) patients and 10 aged controls., Results: Bcl-2 immunoreactivity was widely distributed in neurons, concentrating predominantly on the subfields CA3 and CA4 in the stratum pyramidale of hippocampus both in controls and in AD patients. Bcl-2 staining in the labeled neuron was observed mainly in the cytoplasm and neuritic processes, but a few nuclei were also positive. Bcl-2 labeling was also detected in the astrocytes mainly in AD, but sparsely in controls. Double-labeled fluorescence immunocytochemistry showed that most Bcl-2-immunolabeled neurons also exhibited positive staining for ERalpha., Conclusions: Estrogen may function as a regulator of apoptosis to modulate the expression of Bcl-2 in neurons and astrocytes in hippocampus of AD through ERalpha.

Estrogen receptor (ERalpha) plays an important role in the development and progression of breast cancer. Several recent studies have demonstrated that expression of human X-box binding protein 1 (XBP-1) is associated with ERalpha status in breast tumors and overexpressed in a subset of breast tumors. XBP-1 has two splicing variants, which were designated as XBP-1S and XBP-1U, respectively. However, little is known about the expression pattern of XBP-1S and XBP-1U in breast cancer cells and about their roles in ERalpha signaling. In this study, the expression of two splicing forms of XBP-1 was detected in breast cancer cell lines with RT-PCR. Estrogen response element (ERE) -containing luciferase reporter assay was used to determine the effects of XBP-1S and XBP-1U on the transcription activity of ERalpha in MDA-MB-435 breast cancer cells. The result showed that both XBP-1S and XBP-1U enhanced the transcription activity of ERalpha in a hormone-independent and dose-dependent manner and the activity of of XBP-1S is higher than that of XBP-1U. Enhancement of ERE-containing luciferase reporter gene expression by XBP-1S and XBP-1U was dependent on ERalpha. These data suggest that XBP-1S and XBP-1U may play important roles in breast cancer growth and progression through ERalpha signaling.

One third of estrogens in the male are from the testis and the others from outside the testis. Aromatase P450 (CYP19) is an enzyme responsible for the conversion of androgens to estrogens. Estrogens regulate cell function via specific receptors--estrogen receptors (ER) which include ER alpha and ER beta. It has been found that the role of estrogens in male reproduction is complex and important, particularly during the neonatal life. Males lacking ER alpha are completely infertile because ER alpha-induced estrogens regulate the reabsorption of luminal fluid in the head of the epididymis and disruption of this essential function causes sperm to enter the epididymis diluted, rather than concentrated, resulting in infertility. Whereas males lacking aromatase or ER beta are fully fertile. Therefore, it is concluded that ER alpha, but not aromatase or ER beta, is essenitial for normal male fertility.

Objective: To investigate the effect of mutant-type [(12)Asp]K-ras4B gene on the expression of estrogen receptor (ER) alpha and beta and their transcriptional activity as a transcription factor in endometrial carcinoma HEC-1A cell line., Methods: (1) Effect of [(12)Asp]K-ras4B on the expression of ER alpha and beta were determined using Western blot assay. (2) Eukaryotic expression plasmid pGL3-luciferase-ERE containing luciferase report gene and estrogen receptor element (ERE) was constructed, and co-transfected into NIH3T3 and HEC-1A cell lines with pEGFP-N1 to examine the effect of [(12)Asp]K-ras4B on ER transcription that is regulated by estradiol. In addition, they were transfected into pSV5-HER0 (containing full length wide type ERalpha cDNA) and pCMV-rafS621A (inhibiting raf kinase) plasmids to test the effect of [(12)Asp]K-ras4B/raf signal pathway on transcriptional activity of ER proteins., Results: (1) Protein level of ERs expressed in pcDI transfected control cells was low while it was increased for 3.6-fold (97 +/- 25, 349 +/- 67, P < 0.01) and 1.9-fold (128 +/- 37, 349 +/- 30, P < 0.05) in ERalpha and ERbeta, respectively, in pcDI-[(12)Asp]K-ras4B NIH3T3 cells after transfection. (2) In pcDI-[(12)Asp]K-ras4B NIH3T3 cells, the ratios for ERalpha and and ERbeta levels before transfection of rafS621A plasmids to that after the transfection, were 2.4:1 (724 +/- 45, 310 +/- 46, P < 0.05) and 1.8:1 (493 +/- 20, 284 +/- 20, P < 0.01), respectively; In HEC-1A cells, these ratios were 2.1:1 (566 +/- 22, 279 +/- 30, P < 0.01) and 2.4:1 (405 +/- 33, 165 +/- 15, P < 0.01), respectively. (3) In low serum (2%) culture condition, estradiol (E(2)) stimulated luciferase activity with an increase of 13-fold (130 +/- 42, 1681 +/- 242, P < 0.01) in pcDI-[(12)Asp] K-ras4B NIH3T3 cells, 19-fold (141 +/- 39, 2644 +/- 331, P < 0.001) in HEC-1A cells, respectively, when compared with those in the absence of E(2). (4) In pSV5-HER0 transfected pcDI-[(12)Asp] K-ras4B NIH3T3 cells and HEC-1A cells, compared to the untransfected cells, the ER transcriptional activity in the transfected cells increased markedly. The luciferase activity was increased for 8-fold (1048 +/- 91, 8099 +/- 452, P < 0.01) and 6-fold (2148 +/- 259, 12,705 +/- 2670, P < 0.001), respectively. rafS621A mutant had suppressive effects on luciferase activities in HEC-1A cells and pcDI-[(12)Asp]K-ras4B NIH3T3 cells. The ratio of luciferase activities in pcDI-[(12)Asp]K-ras4B NIH3T3 and HEC-1A cells, before and after transfection was 7.8:1 (1184 +/- 168, 152 +/- 27, P < 0.05) and 6.4:1 (1949 +/- 212, 304 +/- 60, P < 0.01), respectively., Conclusions: (1) [(12)Asp]K-ras4B can enhance the expression of ERalpha and beta proteins. This may be correlated with [(12)Asp]K-ras4B/raf signaling pathway. (2) The effect of mutant-type [(12)Asp]K-ras4B gene on ERs transcriptional activity in HEC-1A cells appears to need E(2).

Immunocytochemical localization were investigated in the nervous system, wheel organ, Hatschek's pit and gonads of amphioxus using polyclonal antibodies against estrogen receptor-alpha and beta. The results revealed that ER-alpha and beta protein distributed in the above regions in larvae and adult at different developmental stages of both sexes. A major ER-alpha were expressed within nucleus of nerve cells, a few expressed in the cytoplasm and process as well as fiber of nerve cells in the forebrain, midbrain, hindbrain and nerve tube, while ER-beta were detected in the cytoplasm or on the cellular membrane, a few were within nucleus. ER-alpha immunopositive material distributed mainly in the nucleus of epithelial cells in the second layer of Hatschek's pit, a few were in the cytoplasm of upper layer epithelial cells, while ER-beta distributed in the nucleus of upper layer. In gonads, ER-alpha were distributed in the cytoplasm and nucleolus of oogonia and oocyte of small growth stage, germinal vesivle(nucleus) showed immunonegative reaction. In the large growth stage, strong immunopositive reaction were showed in nuclear membrane and nucleolus of oocyte, and within nucleus of mature egg cell in the mature stage. ER-beta immunopositive material distributed in the cytoplasm of oogonia and early oocytes as well as egg envelope of mature egg cell, the germinal vesicle showed immunonegative reaction. In testis, both ER subtype were localized in the cytoplasm of spermatogonia, primary and second spermatocyte as well as Sertoli cell, and within nucleus of spermatid cell, while spermatozoa showed immunonegative reaction. On the other hand, the results of double staining revealed that ER-alpha and beta numerously coexisted in a same cell, and fewly expressed in different cells in above regions. It is found for the first time that both estrogen receptor subtype, which mediated the regulate role of estrogen to neuroendocrine tissues in amphioxus, distributed extensively in amphiuoxus. The different localization of ER-alpha and beta receptor in the target cells suggests that the mediating estrogen signal line and the mechanism of gene transcription may be different.

The estrogen receptor (ERalpha) is a member of a large superfamily of nuclear receptors that regulates the transcription of estrogen-responsive genes. Several recent studies have demonstrated that human X-box binding protein 1 (XBP-1) mRNA expression is associated with ERalpha status in breast tumors. More recently, two forms of XBP-1 were identified due to their unique splicing. The two splicing variants of XBP-1 were designated XBP-1S and XBP-1U, respectively. In this study, the coding sequences of XBP-1S and XBP-1U were cloned respectively into the expression vector pcDNA3 harboring FLAG epitope, generating the recombinant plasmids pcDNA3-FLAG-XBP-1S and pcDNA3-FLAG-XBP-1U. Western blot analysis showed that both XBP-1S and XBP-1U were expressed in mammalian cells. To determine the effects of XBP-1S and XBP-1U on the transcriptional activity of ERalpha, MDA-MB-231 breast cancer cells were cotransfected with the expression vectors for ERalpha and either pcDNA3-FLAG-XBP-1S or pcDNA3-FLAG-XBP-1U. The results indicated that XBP-1S and XBP-1U enhanced ERalpha-mediated transcriptional activities in a hormone-independent manner. GST pull-down assay showed that both XBP-1S and XBP-1U interacted with ERalpha. These data suggest that XBP-1S and XBP-1U may play an important role in breast cancer growth and progression through ERalpha signaling.

Objective: AIM To establish a drug screening model based on transcriptional regulation of estrogen responsive element (ERE) and use it to screen compounds for discovering new ligands of estrogen receptor (ER) subtypes., Method: A recombinant reporter vector pERE-TAL-SEAP was constructed by inserting a synthetic sequence composed of five tandem copies of EREs upstream of promoter of the reporter vector pTAL-SEAP. The pERE-TAL-SEAP and the internal control plasmid pCMV were transiently co-transfected into Hela cells expressing ER subtype or ER subtype, and the effects of pure ER agonists 17estradiol, phytoestrogen genistein and pure ER antagonist ICI182, 780 on reporter gene SEAP expression were observed., Result: In the Hela cells expressing ER alpha or ER beta subtype, the expression of SEAP gene were induced in a dose dependent manner by 17-estrodiol with a maximal effect at approximately 10 nmol.L-1 and with EC50 of (80.58 +/- 8.51) pmol.L-1 and (103.90 +/- 5.29) pmol.L-1, respectively, so done by phytoestrogen genistein with a maximal effect at 1 mumol.L-1 and with EC50 of (10.86 +/- 0.75) nmol.L-1 and (39.38 +/- 2.26) nmol.L-1, respectively. The maximal level induced by estrodiol and genistein were about 7-14 fold higher than that of vehicle. The pure antiestrogen ICI182, 780 at concentration of 1 mumol.L-1 completely blocked the inductions of 17-estrodiol and genistein., Conclusion: The cellular drug screening model can be established by transfecting reporter vector pERE-TAL-SEAP in Hela cell lines expressing ER alpha or ER beta. The cell lines can be used to screen compounds with estrogenicity by testing SEAP activity in the culture media of cells growing in microtitier wells. The system should provide an efficient model for screening and analyzing the activity of large numbers of ligands of ER.

Objective: To determine the expression and location of estrogen and progesterone receptor isoforms in the endometrium after abortion and discuss the relationship between the changes of isoforms and prolonged uterine bleeding after medical abortion., Methods: 45 endometrium biopsy were obtained from 15 women with prolonged bleeding after medical abortion, 15 women with prolonged bleeding after vacuum aspiration and 15 without prolonged bleeding after vacuum aspiration as control group. Immunohistochemical staining intensity of ER alpha, ER beta, PR-A and PR-B in endometrium were compared among these groups, as medical abortion group, surgical abortion group and control group., Results: The endometrial ER alpha, ER beta, PR-A and PR-B H-Scores of medical abortion group were 166.7 (88.5), 161.5 (84), 180.0 (109) and 136.5 (119.0), The endometrial ER alpha, ER beta, PR-A and PR-B H-Scores of control group were 306.7 (45.9), 230.0 (75.0), 224.2 (57) and 389.6 (106.6). The endometrial ER alpha, ER beta, PR-A and PR-B H-Scores of surgical abortion group were 109.4 (114.6), 225.0 (120.0), 175.5 (225) and 50.0 (89.0). The endometrial ER alpha, ER beta, PR-B H-Scores of medical abortion group were significantly lower than those of control group. And in surgical abortion group ER alpha and PR-B H-Scores were significantly lower than those in control group. But there was no significant difference between ER beta H-Scores of surgical abortion group and control group. There were no significant differences of ER and PR isoforms level ratios of endometrial glands to endometrial stroma among three groups., Conclusions: The decreased ER alpha, ER beta and PR-B levels in the human endometrium may be related to prolonged uterine bleeding after medical abortion by mifepristone accompanied with misopristol. The endometrial ER alpha, PR-B decreased in women after vacuum aspiration abortion may be related to abnormal prolonged uterine bleeding. The changes of endometrial ER, PR isoforms in glands and stroma were simultaneous after medical abortion and surgical abortion.

Aim: To establish a sensitive and efficient reporter gene based screening model and use it to screen compounds for discovering new ligands of estrogen receptor alpha subtype., Methods: A recombinant Epstein-Barr virus episomal vector (pMT/ERE-CAT) was constructed by inserting a synthetic sequence composed of five estrogen response elements upstream of promoter and a chloramphenicol acetyltransferase (CAT) gene downstream of promoter. pMT/ERE-CAT was transfected into HepG2 cells expressing estrogen receptor alpha subtype (ER +HepG2). Hygromycin (200 micrograms.mL-1) was added 48 h after transfection for selection. One stably transfected clone was isolated and used to screen compounds for activity of stimulating CAT gene expression using colorimetric CAT assay., Results: In the ER +HepG2 cells, the expression of CAT gene was induced by estradiol. A dose-dependent expression of CAT gene with half-maximal induction at 0.07 nmol.L-1 was observed. The ER +HepG2 cell was used to screen compounds for activity of stimulating CAT gene expression. Resveratrol was found to produce a maximal level of induction (1.75 times of estradiol). In vitro radiation survival experiment showed that the radioprotection activity of resveratrol (D0 = 3.18 Gy) is stronger than that of estradiol (D0 = 2.59 Gy)., Conclusion: Vector pMT/ERE-CAT was used to generate stably transfected ER +HepG2 cell lines. The cell lines can be used to screen compounds for estrogen activity by testing extracts of cells grown in microtiter wells directly using colorimetric CAT assay. This system should provide an efficient method for screening and analyzing the activity of large numbers of ligands of estrogen receptor.