Objective: To study the effect of Taoren Quyu Decoction (TQD) on endometrial cells in patients with endometriosis (EMs) and EMs in rats. Methods: A total of 60 female Wistar rats were randomly divided into 4 groups, n...Objective: To study the effect of Taoren Quyu Decoction (TQD) on endometrial cells in patients with endometriosis (EMs) and EMs in rats. Methods: A total of 60 female Wistar rats were randomly divided into 4 groups, namely, normal group, model group, positive group and TQD group, each group having 15 rats. Except the normal group, EMs model was established in the other three groups by transplanting the rat autologous endometrium. After 4 weeks of intragastric administration, blood, eutopic and ectopic endometrial tissues of rats in each group were collected to detect the serum levels of estrogen (E2), cancer antigen 125 (CA125), endometrial antibody (EMA13), and expressions of microvessel density (MVD), vascular endothelial growth factor (VEGF) and angiopoietin (Ang-2). The volume of endometriosis cyst was determined simultaneously. For the in vitro culture of human endometrial cells, 4 groups, namely, normal group, model group, positive group and TQD group were used. The positive group and TQD group were treated with danazol and TQD respectively. Then 24 h after the treatment, the expressions of survivin and tumor suppressor gene (p53) of each group were detected. Results: The volumes of the endometriosis cysts in the positive group and the TQD group were significantly reduced compared with the model group (P<0.05). The serum levels of E2, CA125 and EMAb, and the expressions of MVD. VEGF and Ang-2 in the model group were significantly increased compared with the normal group (P<0.0.5): while they were all significantly reduced in the positive group and TQD group (P<0.05). Compared with the normal group, the expression of survivin in the model group was significantly up regulated (/1/40.05), and expression of p53 was significantly reduced (P<0.05); compared with the model group, the expressions of survivin in the positive and TQD groups were significantly decreased (P<0.05), and expression of p53 was significantly up-regulated (P<0.05). The difference between positive group and TQD group was not statistically significant (P>0.05). Conclusions: TQD has a significant anti-EMs effect, and its mechanism of action may be related to anti-angiogenesis and promoting apoptosis of ectopic endometrial cell.展开更多
In order to investigate the mode of action ofYimu Shenghuasan preparation in endometrial cells of dairy cows, the primary cultured endometrial cells in cows were isolated and the inflammatory models were made by lipop...In order to investigate the mode of action ofYimu Shenghuasan preparation in endometrial cells of dairy cows, the primary cultured endometrial cells in cows were isolated and the inflammatory models were made by lipopolysaccharide （LPS） induction. The inflammatory cells were treated with gradient concentration of herbal medicine preparation, Yimu Shenghuasan for 48 and 72 h. The expression of cytochrome P450 （CYP450） was detected by Western blot. The amounts of IgG and lgA in sera were also detected in the endometritis of dairy cows. The expression level of CYP450 in the endometrial cells of dairy cow was increased gradually, and the amounts of IgG, IgA were increased significantly as compared with those in the control group. The expression level of CYP450 in the inflammatory cells was increased significantly in the treatment of 2 000 μg mL^-1 of Yimu Shenghuasan after 48 h of treatment.展开更多
Background:Endometriosis(EM)is a complex benign gynecological disease,but it has malignant biological behavior and can invade any part of the body.Clinical manifestations include pelvic pain,dysmenorrhea,infertility,p...Background:Endometriosis(EM)is a complex benign gynecological disease,but it has malignant biological behavior and can invade any part of the body.Clinical manifestations include pelvic pain,dysmenorrhea,infertility,pelvic nodules,and masses.Our previous study successfully detected circulating endometrial cells(CECs)in the peripheral blood of patients with EM.The purpose of this study is to overcome the limitation of cell size in the previous microfluidic chip method,to further accurately capture CECs,understand the characteristics of these cells,and explore the relationship between CECs and the clinical course characteristics of patients with EM.Methods:Human peripheral venous blood used to detect CECs and circulating vascular endothelial cells(CVECs)was taken from EM patients(n=34)hospitalized in the Peking University People’s Hospital.We used the subtraction enrichment and immunostaining fluorescence in situ hybridization(SE-iFISH)method to exclude the interference of red blood cells,white blood cells,and CVECs,so as to accurately capture the CECs in the peripheral blood of patients with EM.Then we clarified the size and ploidy number of chromosome 8 of CECs,and a second grouping of patients was performed based on clinical characteristics to determine the relationship between CECs and clinical course characteristics.Results:The peripheral blood of 34 EM patients and 12 non-EM patients was evaluated by SE-iFISH.Overall,34 eligible EM patients were enrolled.The results showed that the detection rates of CECs were 58.8%in EM patients and 16.7%in the control group.However,after classification according to clinical characteristics,more CECs could be detected in the peripheral blood of patients with rapidly progressive EM,with a detection rate of 94.4%(17/18).In total,63.5%(40/63)of these cells were small cells with diameters below 5μm,and 44.4%(28/63)were aneuploid cells.No significant association was found between the number of CECs and EM stage.Conclusion:The number and characteristics of CECs are related to the clinical course characteristics of patients with EM,such as pain and changes in lesion size,and may be used as biomarkers for personalized treatment and management of EM in the future.展开更多
The development of receptive endometrium(RE) from pre-receptive endometrium(PE) for successful embryo implantation is a complex dynamic process in which the morphology and physiological states of the endometrial epith...The development of receptive endometrium(RE) from pre-receptive endometrium(PE) for successful embryo implantation is a complex dynamic process in which the morphology and physiological states of the endometrial epithelium undergo a series of significant changes, including cell proliferation and apoptosis. However, the molecular mechanisms are not yet fully understood. In this study, a higher circRNA3669 level was observed in PE than in RE of goats. Functional assays revealed that this overexpression promoted the proliferation of goat endometrial epithelial cells(GEECs) by activating the expression of genes related to the PI3K/AKT-mTOR and MAPK pathways,thereby inhibiting apoptosis in vitro. Furthermore, circRNA3669 functioned as a competing endogenous RNA(ceRNA) to upregulate Reticulocalbin-2(RCN2) expression at the post-transcriptional level by interacting with and downregulating miR-26a in GEECs. In addition, RCN2, which is highly expressed in the PE of goats, was found to be regulated by β-estradiol(E2) and progesterone(P4). Our results demonstrated that RCN2 also affected the key proteins PI3K, AKT, mTOR, JNK, and P38 in the PI3K/AKT-mTOR and MAPK pathways, thereby facilitating GEECs proliferation and suppressing their apoptosis in vitro. Collectively, we constructed a new circRNA3669-miR-26aRCN2 regulatory network in GEECs, which further provides strong evidence that circRNA could potentially play a crucial regulatory role in the development of RE in goats.展开更多
Background： Circulating endometrial cells （CECs） have been reported to be present in the peripheral blood of women with endometriosis （EM）, providing clear and specific evidence of the presence of ectopic lesions...Background： Circulating endometrial cells （CECs） have been reported to be present in the peripheral blood of women with endometriosis （EM）, providing clear and specific evidence of the presence of ectopic lesions. In this study, we established a method with a high detection rate of CECs, assessed the diagnostic value of CECs for EM and compared with serum CA125, and proposed a hypothesis for the pathogenesis of EM from the new perspective of CECs. Methods： The participants were enrolled prospectively from October 2015 to July 2016. The peripheral blood samples were collected from 59 participants, and the blood cells were isolated for immunofluorescence staining via microfluidic chips. The cells that were positive for vimentin/cytokeratin and estrogen/progesterone receptor and negative for CD45 were identified as CECs. The serum CA125 level was tested with electrochemiluminescence immunoassay. Results： The detection rate of CECs reached 89.5% （17/19） in the EM group, which was significantly higher than that of the control group （15.0% [6/40], P 〈 0.001） and was independent of menstrual cycle phases. Furthermore, a positive CEC assay detected 4/5 cases of Stage Ⅰ–Ⅱ EM. In contrast, a positive CA125 test had limited value in detecting EM （13/19, 68.4%） and detected only one case of Stage Ⅰ–Ⅱ EM. Conclusion： CECs are promising biomarkers for EM with great potential for a noninvasive diagnostic assay.展开更多
BACKGROUND Type 1 diabetes(T1D),a chronic metabolic and autoimmune disease,seriously endangers human health.In recent years,mesenchymal stem cell(MSC)transplantation has become an effective treatment for diabetes.Mens...BACKGROUND Type 1 diabetes(T1D),a chronic metabolic and autoimmune disease,seriously endangers human health.In recent years,mesenchymal stem cell(MSC)transplantation has become an effective treatment for diabetes.Menstrual bloodderived endometrial stem cells(MenSC),a novel MSC type derived from the decidual endometrium during menstruation,are expected to become promising seeding cells for diabetes treatment because of their noninvasive collection procedure,high proliferation rate and high immunomodulation capacity.AIM To comprehensively compare the effects of MenSC and umbilical cord-derived MSC(UcMSC)transplantation on T1D treatment,to further explore the potential mechanism of MSC-based therapies in T1D,and to provide support for the clinical application of MSC in diabetes treatment.METHODS A conventional streptozotocin-induced T1D mouse model was established,and the effects of MenSC and UcMSC transplantation on their blood glucose and serum insulin levels were detected.The morphological and functional changes in the pancreas,liver,kidney,and spleen were analyzed by routine histological and immunohistochemical examinations.Changes in the serum cytokine levels in the model mice were assessed by protein arrays.The expression of target proteins related to pancreatic regeneration and apoptosis was examined by western blot.RESULTS MenSC and UcMSC transplantation significantly improved the blood glucose and serum insulin levels in T1D model mice.Immunofluorescence analysis revealed that the numbers of insulin+and CD31+cells in the pancreas were significantly increased in MSC-treated mice compared with control mice.Subsequent western blot analysis also showed that vascular endothelial growth factor(VEGF),Bcl2,Bcl-xL and Proliferating cell nuclear antigen in pancreatic tissue was significantly upregulated in MSC-treated mice compared with control mice.Additionally,protein arrays indicated that MenSC and UcMSC transplantation significantly downregulated the serum levels of interferonγand tumor necrosis factorαand upregulated the serum levels of interleukin-6 and VEGF in the model mice.Additionally,histological and immunohistochemical analyses revealed that MSC transplantation systematically improved the morphologies and functions of the liver,kidney,and spleen in T1D model mice.CONCLUSION MenSC transplantation significantly improves the symptoms in T1D model mice and exerts protective effects on their main organs.Moreover,MSC-mediated angiogenesis,antiapoptotic effects and immunomodulation likely contribute to the above improvements.Thus,MenSC are expected to become promising seeding cells for clinical diabetes treatment due to their advantages mentioned above.展开更多
Objective To investigate the regulatory effect of multifactor on the matrix metalloproteinases-9 （MMP-9） and the tissue inhibitor of metalloproteinase-1 （TIMP-1） in endometrial stromal cells. Methods The endometri...Objective To investigate the regulatory effect of multifactor on the matrix metalloproteinases-9 （MMP-9） and the tissue inhibitor of metalloproteinase-1 （TIMP-1） in endometrial stromal cells. Methods The endometrial stromal cells separated from the proliferative endometrial tissues were incubated with medium alone, 17-β estradiol （E2,10^-8 mol/L）, medroxyprogesterone acetate （MPA, 10^-6 mol/L）, E2（10^-8 mol/L）＋MPA （10^-6 mol/L）, E2 （10^-8 mol/L）＋MPA （10^-6 mol/L）＋RU486 （10^-5 mol/L） or HB-EGF （10 ng/ml） for 48 h respectively. The expressions of MMP-9 and TIMP-1 were detected by in situ hybridization, immunocytochemistry, reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting. Results Compared with control group [mRNA, 0. 729 ± 0. 090 （MMP-9） and 1.056± 0.154 （TIMP-1）; protein, 0.545 ±0.086 （MMP-9） and 0.745 ±0.154 （TIMP-1）], expressions of MMP-9 and TIMP-1 in E2 alone, progestin alone or E2 combined with progestin group were respectively：mRNA, 0.413 ± 0.069, 0.402 ± 0.073 and 0.407 ± 0.039; 0.487 ± 0.093, 0.503 ± 0.093 and 0.468 ± 0.075：protein, 0.294 ± 0.076, 0.331 ±0.064 and 0.265 ±0.049; 0.425 ±0.085, 0.397 ±0.065 and 0.435 ± 0.099. RU486 weakened the expression level of down-regulation, while HB-EGF elevated the level of MMP-9 and TIMP-1 after 48 h treatment （mRNA, 0.955 ± 0.068 and 1.396 ± 0.238; protein, 0. 780 ± 0.109 and 0.985 ± 0.165）. Conclusions 1） Both E2 and progestin can down-regulate the expressions of MMP-9 and TIMP-1 in endometrial stromal cells, but RU486 can inhibit the effect. 2） HB-EGF can elevate the level of MMP-9 and TIMP-1. 3） E2, progestin and HB-EGF have effect on the ratio of MM-P/TIMP-1.展开更多
Objective： To observe the effect of the artesunate （ART） on cellular proliferation in vitro, to search for the possible anti-tumor mechanism of ART on endometrial carcinoma at the molecular level and to provide the...Objective： To observe the effect of the artesunate （ART） on cellular proliferation in vitro, to search for the possible anti-tumor mechanism of ART on endometrial carcinoma at the molecular level and to provide the experimental and theoretical foundations for the clinical applications of ART. Methods： The cell proliferation was observed by microscope; MTT was used to examine the effects of ART on proliferation of HEC-1B cells, and flow cytometric analysis was used to detect cell cycle and apoptosis. The human endometrial carcinoma HEC-1B cells were conventionally cultured; ART was administered with a concentration of 40 μg/ml before the total RNA were extracted, mRNA expression of Survivin, Caspase-3, N-Cadherin, E-Cadherin, Fibronectinl and Cox-2 were detected using RT-PCR. Results： ART reduced proliferation in human endometrial carcinoma cell line HEC-1B in a dose- and time-dependent effect. The cells of G0/G1 stage were significantly increased （P〈0.05）, but the cells of G2/M stages were significantly decreased （P〈0.05）, so it has shown that the cell cycle was probably blocked in G0/G1 stage. After intervention with ART at 20 and 80 μg/ml for 48 h, cellular apoptosis rate respectively was （36.42±0.77）% and （11.77±0.58）%, and the difference was statistically significant compared with the control （[6.64±0.191%, P〈0.01）. The expression of Cox-2 mRNA in the ART group was lower than those of control group, yet the expression of Caspase-3 and E-Cadherin mRNA in the ART group was higher than those of control group. Conclusion： ART can inhibit HEC-1B cell growth and proliferation in a dose- and time-dependent manner. Furthermore, ART can induce apoptosis in a dose-dependent manner. ART is able to downregulate Cox-2 mRNA expression and to upregulate E-Cadherin and Caspase-3 mRNA expression. So we can conclude that ART could induce the endometrial carcinoma HEC-1B cell apoptosis and inhibit tumor cell proliferation.展开更多
Objective To investigate differentially expressed genes related to inflammation in endometrial cells induced by Lipopolysaccharide（LPS）. Methods Normal endometrium in the proliferative phase of specimen from 3 cases...Objective To investigate differentially expressed genes related to inflammation in endometrial cells induced by Lipopolysaccharide（LPS）. Methods Normal endometrium in the proliferative phase of specimen from 3 cases for the experiment was collected. The LPS group were treated with 50 μg/ml LPS. Total RNA was extracted using Trizol reagent from cells. RNA quality was assessed by determining the OD260/280 ratio by agarose gel electrophoresis, the chip was scanned by laser scanner. The acquired was analyzed. Results A total of differentially expressed genes were found, these genes were relative to many aspects. Among them, the expression of genes involved in inflammation were up-regulated by LPS, such as overexpression of IL-1β, 8, etc. Conclusion The results indicates that inflammation-related genes may be one of the mechanisms of abnormal uterine bleeding by LPS-induced.展开更多
The purpose of this study was to explore the change of telomerase in passage from human endometrial stromal stem cells isolated from human endometrium.Telomerase activity of cultured endometrial stromal cells was asse...The purpose of this study was to explore the change of telomerase in passage from human endometrial stromal stem cells isolated from human endometrium.Telomerase activity of cultured endometrial stromal cells was assessed at mRNA and protein levels using RT-PCR and immunohistochemistry technique.Telomerase mRNA and protein levels were higher at early passages,and then had a gradually decreased trend of immunoreactive intensity and gradually weakened positive cells with progressive passage in endometrial stromal stem cells.These results suggest that telomerase is contributed to the insenecence of endometrial stem cell.展开更多
[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying bio...[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [Method] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated cells. [Result] The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purity was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [Conclusion] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purity.展开更多
Objective To explore the effect of genistein on proliferation of human endometrial endothelial cells （HEECs） and glandular epithelium. Methods In vitro HEECs and human endometrial cancer-1B cell （HEC-1B ） were cu...Objective To explore the effect of genistein on proliferation of human endometrial endothelial cells （HEECs） and glandular epithelium. Methods In vitro HEECs and human endometrial cancer-1B cell （HEC-1B ） were cultured with 0, 1, 10, 50, 100, and 200 μmol/L of genistein alone or indicated concentrations of genistein combined with 0.2 or 1 nmol/L 17β- estradiol （ 17β-E2）. Cell proliferation was determined by [ 3H ] -thymidine incorporation and cell cycle was measured by flow cytometry. Results After 96 hours of treatment, genistein inhibited the proliferation of HEECs in a dose-dependent manner. The stimulation index reduced from 100% （ without genistein treatment ） to about 1% （ 200 μmol/L genistein ）. HEECs were arrested at G1/0 and G2/M phase when treated with genistein for 96 hours. When the concentration of genistein was 200 μmol/L, the percentages of HEECs at G1/0, G2/M, and S phase were 96. 0%, 2.1%, and 1.9%, respectively. However, when HEECs were treated without genistein, the percentages of HEECs at G1/0, G2/M, and S phase were 76. 7%, 8.5%, and 14. 7%, respectively. 1713-E2 could not influence the effects of genistein on the proliferation of HEECs. Meanwhile, genistein could suppress the proliferation of HEC-1B. If the stimulation index of HEC-IB was defined as 100% when HEC-1B was treated with different doses of 1713-E2 （without genistein）, it was 67%, 19%, as well as 32% when cell was supplemented with 200 μmol/L genistein combined with 0, 0. 2, or 1 nmol/L 17β-E2, respectively. Conclusion Genistein at the concentration of 200 μmol/L can sufficiently inhibit the proliferation of HEECs and endometrial glandular epithelium simultaneously in vitro.展开更多
Primary small cell carcinoma（SCC） is a group of aggressive neoplasms that mainly arise from the lung and digestive tract. Endometrial small cell carcinoma（ESCC） is extremely rare. To our knowledge, less than 90 ca...Primary small cell carcinoma（SCC） is a group of aggressive neoplasms that mainly arise from the lung and digestive tract. Endometrial small cell carcinoma（ESCC） is extremely rare. To our knowledge, less than 90 cases have been reported, and most of these reports were dedicated to describing the clinicopathologic or immunochemical features of ESCC. Herein, we present a new case of ESCC involving a 51-year-old woman and mainly focus on the magnetic resonance imaging（MRI） and positron emission tomography/computed tomography（PET/CT） findings. MRI showed that the uterus was significantly enlarged（11.6 cm × 11.1 cm × 14.4 cm）, and a giant irregular mass（7.5 cm × 8.4 cm × 8.5 cm） was observed in the uterine cavity. The lesion demonstrated an extremely low apparent diffusion coefficient（ADC） value [（0.553±0.088）×10^–3 mm^2/s] and a high FDG uptake value（22.7）. Multiple metastatic lymph nodes（LNs） were identified at different positions, with diameters ranging from 0.3 to 2.8 cm and a maximum standardized uptake value（SUV max） ranging from 6.9 to 19.3.展开更多
BACKGROUND Cervical squamous cell carcinoma(SCC)is the most common type of cervical carcinoma and is generally derived from a precancerous stage called cervical high-grade squamous intraepithelial lesion(HSIL).Usually...BACKGROUND Cervical squamous cell carcinoma(SCC)is the most common type of cervical carcinoma and is generally derived from a precancerous stage called cervical high-grade squamous intraepithelial lesion(HSIL).Usually,the cancer metastasizes through lymphatic or hematogenous dissemination,but rarely spreads upward into the uterus.Here,we report a case of cervical HSIL extending into the endometrium and finally progressing to SCC in the uterine cavity.CASE SUMMARY A 57-year-old postmenopausal woman visited our department and requested a routine cervical check-up.Four years ago,she had undergone a cervical loop electrosurgical excision procedure because of HSIL found during the gynecological examination,and she had not been checked again since.This time,a relapse of the cervical HSIL was diagnosed along with uterine pyometra and endometrial polyps.After 2 wk of antibiotic treatment,a laparoscopic hysterectomy was performed,and the final pathological examination revealed that the cervical HSIL had spread directly upward into the uterine cavity,gradually developing into cervical SCC in the endometrium.CONCLUSION Cervical HSIL/SCC can directly spread upward into the uterus with the most common symptoms of pyometra and cervical stenosis.More attention should be given to the early detection and prevention of this disease.展开更多
s To study the influence of insulin on IGF Ⅰ and IGFBP Ⅰ secretion of the hum an endometrial stromal cells Methods Late proliferative phase endometrial stromal cells were isolated from endometriu m tissues an...s To study the influence of insulin on IGF Ⅰ and IGFBP Ⅰ secretion of the hum an endometrial stromal cells Methods Late proliferative phase endometrial stromal cells were isolated from endometriu m tissues and then cultured for 24 h in Hams F 12 only as a control and in Hams F 12 with different concentrations of estradiol (E2) and insulin (INS) as trea ted groups Simultaneously, the endometrial stromal cells from late secretory phase endometrium were cultured for 24 h in Hams F 12 only as a control and in Hams F 12 supplemented with different concentrations of progesterone (P) and i nsulin as treated groups After 24 h of culturing, the mediums were collected f or either IGF Ⅰ or IGFBP Ⅰ assays Result The concentrations of IGF Ⅰ in medium from cultured endometrial stromal cells i n the proliferative phase were 0 78±0 47 ng/ml in the hormone free control g roup; 1 44±0 59 ng/ml and 1 39± 0 33 ng/ml in 100 pg/ml E2 group and 20 μU /ml INS group, which was higher than that of the control group ( P <0 05 and P <0 01, respectively) The IGF Ⅰ concentration in the 100 μU/ml INS group was 2 03±0 53 ng/ml, which was higher than that of the 20 μU/ml INS group ( P <0 01) Levels of IGF Ⅰ in the 100 pg/ml E2 plus 20 μU/ml INS gro up was 2 18±0 36 ng/ml, which was significantly higher than that of the 20 μU/ml INS and 100 pg/ml E2 group ( P <0 01), but lower than that of the 100 pg/ml E2 p lus 100 μU/ml INS group (3 42±0 75 ng/ml), P <0 01 The concentration o f IGFBP Ⅰ in medium from cultured endometrial stromal cells in the secretory ph a se was 2 50±1 39 ng/ml in the hormone free control group and 5 44±2 09 ng /ml in the 10 pg/ml P group, which was significantly higher than that of the con trol ( P <0 01) IGFBP Ⅰ concentration in 20 μU/ml INS group was 0 1 6±0 58 ng/ml, which was lower compared with control, but higher compared with the 100 μU/ml INS group ( P <0 01) The level of IGFBP Ⅰ in the 10 ng/ml P plu s 20 μU/ml INS group was 2 10±1 17 ng/ml, lower compared with the 10 ng/ml P gr oup, but higher compared with the 10 pg/ml P plus 100 μU/ml INS group, P <0 01 Conclusions Insulin can stimulate basal (without hormone) and E2 stimulated IGF Ⅰ secreti on in cultured stromal cells from human late proliferative endometrium in a dose dependent manner Insulin can suppress basal (without hormone) and P stimula ted IGFBP Ⅰ secretions in cultured stromal cells from human secretory endomet rium in a dose dependent manner展开更多
Objective：Bovine endometritis is one of the most common reproductive disorders in cattle.The aim of this study was to investigate the anti-inflammation potential of punicalagin in lipopolysaccharide（LPS）-induced bo...Objective：Bovine endometritis is one of the most common reproductive disorders in cattle.The aim of this study was to investigate the anti-inflammation potential of punicalagin in lipopolysaccharide（LPS）-induced bovine endometrial epithelial cells（bE ECs）and to uncover the underlying mechanisms.Methods：bE ECs were stimulated with different concentrations（1,10,30,50,and 100μg/ml）of LPS for 3,6,9,12,and 18 h.MTT assay was used to assess cell viability and to identify the conditions for inflammatory injury and effective concentrations of punicalagin.Quantitative real-time polymerase chain reaction（q RT-PCR）was used to assess gene expression of pro-inflammatory cytokines.Western blotting was used to assess levels of inflammation-related proteins.Results：Treatment of b EECs with 30μg/ml LPS for 12 h induced cell injury and reduced cell viability.Punicalagin（5,10,or 20μg/ml）pretreatment significantly decreased LPS-induced productions of interleukin（IL）-1β,IL-6,IL-8,and tumor necrosis factor-α（TNF-α）in bE ECs.Molecular research showed that punicalagin inhibited the activation of the upstream mediator nuclear factor-κB（NF-κB）by suppressing the production of inhibitorκBα（IκBα）and phosphorylation of p65.Results also indicated that punicalagin can suppress the phosphorylation of mitogen-activated protein kinases（MAPKs）including p38,c-Jun N-terminal kinase（JNK）,and extracellular signal-regulated kinase（ERK）.Conclusions：Punicalagin may attenuate LPS-induced inflammatory injury and provide a potential option for the treatment of dairy cows with Escherichia coli endometritis.展开更多
Extracellular vesicles from highly metastatic tumor cells have been shown to mediate epithelial-mesenchymal transition(EMT)-related events in recipient cells.In endometrial epithelial cells,EMT processes are known to ...Extracellular vesicles from highly metastatic tumor cells have been shown to mediate epithelial-mesenchymal transition(EMT)-related events in recipient cells.In endometrial epithelial cells,EMT processes are known to be involved in the development of adenomyosis.We aimed to investigate whether adenomyosis-derived extracellular vesicles(AMEVs)are able to induce an EMT process in endometrial epithelial cells.In this study,AMEVs were isolated from patients with adenomyosis and characterized by transmission electron microscopy,Western blot,and nanoparticle tracking.Primary endometrial epithelial cells(EECs)were derived from normal endometrium tissues from patients with leiomyoma and co-cultured with AMEVs in vitro.AMEV uptake was examined by fluorescence confocal microscopy.The invasion of EECs was confirmed by Transwell assay.Immunohistochemistry,Western blot,and qRT-PCR were performed on EECs to illustrate the expression levels of cytokeratin 19,E-cadherin,vimentin,and zinc finger E-boxbinding homeobox 1(ZEB1).The results indicated that the cellular fluorescence intensity gradually increased after 48 h of co-culture,but decreased after 72 h.After co-culturing with AMEVs for 72 h,EECs expressed significantly lower levels of cytokeratin 19 and E-cadherin,and significantly higher levels of vimentin and ZEB1.Together these results demonstrated that AMEVs induce an EMT process and enhance the invasion of EECs.These changes may contribute to the pathogenesis and progression of adenomyosis.展开更多
Objective:This study aimed to investigate the differentiation of human adipose-derived stem cells(hASCs)into endometrial epithelial cells(EECs)under certain induction conditions and to a further step provide a promisi...Objective:This study aimed to investigate the differentiation of human adipose-derived stem cells(hASCs)into endometrial epithelial cells(EECs)under certain induction conditions and to a further step provide a promising approach for ASCs in clinical practice to the treatment of severe intrauterine adhesion.Methods:Four groups of hASCs were separately cultured as follows:in Group 1,hASCs were cultured in a control medium(5%fetal bovine serum[FBS]+α-minimum Eagle’s medium[α-MEM]);in Group 2,hASCs were cultured in an induction medium(5%FBS+α-MEM+[1×10-7 mol/L 17β-estradiol]+10 ng/mL transforming growth factorβ1[TGF-β1]+10 ng/mL epidermal growth factor[EGF]+10 ng/mL platelet-derived growth factor BB[PDGF-BB]);in Group 3,hASCs and human endometrium cells(hEMCs)were cocultured in the control medium;and in Group 4,hASCs and hEMCs were cocultured in the induction medium.Results:When cocultured with hEMCs,the morphology of hASCs became similar with EECs,and the addition of factors such as EGF,TGFβ,PDGF-BB,and 17β-estradiol promoted differentiation.This study,for the first time,demonstrated estrogen receptor(ER)αand ERβexpression in hASCs and preliminarily explored changes in ERα,ERβ,β-catenin,and H19 mRNA expression during hASC differentiation.Furthermore,we concluded that H19 mRNA expression was negatively correlated with differentiation,which is seemingly related to the estrogen signaling pathway.Conclusions:hASCs revealed the potential for differentiating to EECs when cocultured with hEMCs.展开更多
Endometrial stromal cell decidualization is a crucial step in endometrial remodeling during pregnancy.Decidualization is controlled by orchestrated ovarian hormones,followed by the activation of various downstream sig...Endometrial stromal cell decidualization is a crucial step in endometrial remodeling during pregnancy.Decidualization is controlled by orchestrated ovarian hormones,followed by the activation of various downstream signaling pathways.Accumulating evidence has shown multiple functions of decidualized endometrial stromal cells during embryo implantation,including tissue remodeling,antioxidative stress,angiogenesis,and immune tolerance.The distinct secretomes of decidualized stromal cells also reveal their intensive interactions with epithelial,endothelial,and immune cells.However,aberrant decidualization leads to pregnancy failures,such as recurrent pregnancy loss and repeated implantation failure.This review aimed to provide an overview of the molecular mechanisms underlying the divergent functions of decidualized endometrial stromal cells and their potential clinical applications.Moreover,the use of single-cell RNA sequencing data further enhances our understanding of these biological processes.This review discusses decidualization-related signaling pathways that serve as potential therapeutic targets for treating implantation failure in in vitro fertilization and provides novel approaches to investigate the underlying causes of female infertility.展开更多
Numerous studies have shown aberrant immune cell function in endometriosis,including T cells,B cells,natural killer cells,and macrophages(MΦ).These alterations are thought to be induced by various mechanisms that pro...Numerous studies have shown aberrant immune cell function in endometriosis,including T cells,B cells,natural killer cells,and macrophages(MΦ).These alterations are thought to be induced by various mechanisms that promote the disease.Regulatory T cells(Tregs)may account for a decreased ability of newly recruited leukocytes to initiate effective immune responses against viable endometrial fragments,permitting their survival.Tregs differentiate during the development of endometriosis,which confer immunosuppression or play other roles in disease progression.In this review,we provide an overview of the regulation and roles of Tregs in endometriosis.These data provide further scientific evidence for the altered immune response in endometriosis,which could be a potential target in the treatment of endometriosis.This review could create new diagnostic strategies and effective immune-targeted therapies for this highly prevalent disease.Recent progress in the field indicates that these goals may be achieved in the future.展开更多
基金supported by Guangdong Provincial Medical Research Fund Project(A2011345)
文摘Objective: To study the effect of Taoren Quyu Decoction (TQD) on endometrial cells in patients with endometriosis (EMs) and EMs in rats. Methods: A total of 60 female Wistar rats were randomly divided into 4 groups, namely, normal group, model group, positive group and TQD group, each group having 15 rats. Except the normal group, EMs model was established in the other three groups by transplanting the rat autologous endometrium. After 4 weeks of intragastric administration, blood, eutopic and ectopic endometrial tissues of rats in each group were collected to detect the serum levels of estrogen (E2), cancer antigen 125 (CA125), endometrial antibody (EMA13), and expressions of microvessel density (MVD), vascular endothelial growth factor (VEGF) and angiopoietin (Ang-2). The volume of endometriosis cyst was determined simultaneously. For the in vitro culture of human endometrial cells, 4 groups, namely, normal group, model group, positive group and TQD group were used. The positive group and TQD group were treated with danazol and TQD respectively. Then 24 h after the treatment, the expressions of survivin and tumor suppressor gene (p53) of each group were detected. Results: The volumes of the endometriosis cysts in the positive group and the TQD group were significantly reduced compared with the model group (P<0.05). The serum levels of E2, CA125 and EMAb, and the expressions of MVD. VEGF and Ang-2 in the model group were significantly increased compared with the normal group (P<0.0.5): while they were all significantly reduced in the positive group and TQD group (P<0.05). Compared with the normal group, the expression of survivin in the model group was significantly up regulated (/1/40.05), and expression of p53 was significantly reduced (P<0.05); compared with the model group, the expressions of survivin in the positive and TQD groups were significantly decreased (P<0.05), and expression of p53 was significantly up-regulated (P<0.05). The difference between positive group and TQD group was not statistically significant (P>0.05). Conclusions: TQD has a significant anti-EMs effect, and its mechanism of action may be related to anti-angiogenesis and promoting apoptosis of ectopic endometrial cell.
基金supported by the National Technology Support Project of China(2006BAD04A10-4)
文摘In order to investigate the mode of action ofYimu Shenghuasan preparation in endometrial cells of dairy cows, the primary cultured endometrial cells in cows were isolated and the inflammatory models were made by lipopolysaccharide （LPS） induction. The inflammatory cells were treated with gradient concentration of herbal medicine preparation, Yimu Shenghuasan for 48 and 72 h. The expression of cytochrome P450 （CYP450） was detected by Western blot. The amounts of IgG and lgA in sera were also detected in the endometritis of dairy cows. The expression level of CYP450 in the endometrial cells of dairy cow was increased gradually, and the amounts of IgG, IgA were increased significantly as compared with those in the control group. The expression level of CYP450 in the inflammatory cells was increased significantly in the treatment of 2 000 μg mL^-1 of Yimu Shenghuasan after 48 h of treatment.
基金supported by grants from the National Key Research and Development Program of China(No.2022YFC2704000)the National Natural Science Foundation of China(No.81971360).
文摘Background:Endometriosis(EM)is a complex benign gynecological disease,but it has malignant biological behavior and can invade any part of the body.Clinical manifestations include pelvic pain,dysmenorrhea,infertility,pelvic nodules,and masses.Our previous study successfully detected circulating endometrial cells(CECs)in the peripheral blood of patients with EM.The purpose of this study is to overcome the limitation of cell size in the previous microfluidic chip method,to further accurately capture CECs,understand the characteristics of these cells,and explore the relationship between CECs and the clinical course characteristics of patients with EM.Methods:Human peripheral venous blood used to detect CECs and circulating vascular endothelial cells(CVECs)was taken from EM patients(n=34)hospitalized in the Peking University People’s Hospital.We used the subtraction enrichment and immunostaining fluorescence in situ hybridization(SE-iFISH)method to exclude the interference of red blood cells,white blood cells,and CVECs,so as to accurately capture the CECs in the peripheral blood of patients with EM.Then we clarified the size and ploidy number of chromosome 8 of CECs,and a second grouping of patients was performed based on clinical characteristics to determine the relationship between CECs and clinical course characteristics.Results:The peripheral blood of 34 EM patients and 12 non-EM patients was evaluated by SE-iFISH.Overall,34 eligible EM patients were enrolled.The results showed that the detection rates of CECs were 58.8%in EM patients and 16.7%in the control group.However,after classification according to clinical characteristics,more CECs could be detected in the peripheral blood of patients with rapidly progressive EM,with a detection rate of 94.4%(17/18).In total,63.5%(40/63)of these cells were small cells with diameters below 5μm,and 44.4%(28/63)were aneuploid cells.No significant association was found between the number of CECs and EM stage.Conclusion:The number and characteristics of CECs are related to the clinical course characteristics of patients with EM,such as pain and changes in lesion size,and may be used as biomarkers for personalized treatment and management of EM in the future.
基金supported by the China Postdoctoral Science Foundation(2019M653776 and 2020M673516)the Natural Science Basis Research Plan in Shaanxi Province of China(2023-JC-QN-0181)+1 种基金the Shaanxi Livestock and Poultry Breeding Double-chain Fusion Key Project,China(2022GD-TSLD-46-0202)the Natural Science Fundation of Tibet Autonomous Region,China(XZ202101ZR0063G)。
文摘The development of receptive endometrium(RE) from pre-receptive endometrium(PE) for successful embryo implantation is a complex dynamic process in which the morphology and physiological states of the endometrial epithelium undergo a series of significant changes, including cell proliferation and apoptosis. However, the molecular mechanisms are not yet fully understood. In this study, a higher circRNA3669 level was observed in PE than in RE of goats. Functional assays revealed that this overexpression promoted the proliferation of goat endometrial epithelial cells(GEECs) by activating the expression of genes related to the PI3K/AKT-mTOR and MAPK pathways,thereby inhibiting apoptosis in vitro. Furthermore, circRNA3669 functioned as a competing endogenous RNA(ceRNA) to upregulate Reticulocalbin-2(RCN2) expression at the post-transcriptional level by interacting with and downregulating miR-26a in GEECs. In addition, RCN2, which is highly expressed in the PE of goats, was found to be regulated by β-estradiol(E2) and progesterone(P4). Our results demonstrated that RCN2 also affected the key proteins PI3K, AKT, mTOR, JNK, and P38 in the PI3K/AKT-mTOR and MAPK pathways, thereby facilitating GEECs proliferation and suppressing their apoptosis in vitro. Collectively, we constructed a new circRNA3669-miR-26aRCN2 regulatory network in GEECs, which further provides strong evidence that circRNA could potentially play a crucial regulatory role in the development of RE in goats.
文摘Background： Circulating endometrial cells （CECs） have been reported to be present in the peripheral blood of women with endometriosis （EM）, providing clear and specific evidence of the presence of ectopic lesions. In this study, we established a method with a high detection rate of CECs, assessed the diagnostic value of CECs for EM and compared with serum CA125, and proposed a hypothesis for the pathogenesis of EM from the new perspective of CECs. Methods： The participants were enrolled prospectively from October 2015 to July 2016. The peripheral blood samples were collected from 59 participants, and the blood cells were isolated for immunofluorescence staining via microfluidic chips. The cells that were positive for vimentin/cytokeratin and estrogen/progesterone receptor and negative for CD45 were identified as CECs. The serum CA125 level was tested with electrochemiluminescence immunoassay. Results： The detection rate of CECs reached 89.5% （17/19） in the EM group, which was significantly higher than that of the control group （15.0% [6/40], P 〈 0.001） and was independent of menstrual cycle phases. Furthermore, a positive CEC assay detected 4/5 cases of Stage Ⅰ–Ⅱ EM. In contrast, a positive CA125 test had limited value in detecting EM （13/19, 68.4%） and detected only one case of Stage Ⅰ–Ⅱ EM. Conclusion： CECs are promising biomarkers for EM with great potential for a noninvasive diagnostic assay.
基金Henan Province Foundation of China,No.202300410307 and No.212102310611Xinxiang City Foundation of China,No.GG2020009.
文摘BACKGROUND Type 1 diabetes(T1D),a chronic metabolic and autoimmune disease,seriously endangers human health.In recent years,mesenchymal stem cell(MSC)transplantation has become an effective treatment for diabetes.Menstrual bloodderived endometrial stem cells(MenSC),a novel MSC type derived from the decidual endometrium during menstruation,are expected to become promising seeding cells for diabetes treatment because of their noninvasive collection procedure,high proliferation rate and high immunomodulation capacity.AIM To comprehensively compare the effects of MenSC and umbilical cord-derived MSC(UcMSC)transplantation on T1D treatment,to further explore the potential mechanism of MSC-based therapies in T1D,and to provide support for the clinical application of MSC in diabetes treatment.METHODS A conventional streptozotocin-induced T1D mouse model was established,and the effects of MenSC and UcMSC transplantation on their blood glucose and serum insulin levels were detected.The morphological and functional changes in the pancreas,liver,kidney,and spleen were analyzed by routine histological and immunohistochemical examinations.Changes in the serum cytokine levels in the model mice were assessed by protein arrays.The expression of target proteins related to pancreatic regeneration and apoptosis was examined by western blot.RESULTS MenSC and UcMSC transplantation significantly improved the blood glucose and serum insulin levels in T1D model mice.Immunofluorescence analysis revealed that the numbers of insulin+and CD31+cells in the pancreas were significantly increased in MSC-treated mice compared with control mice.Subsequent western blot analysis also showed that vascular endothelial growth factor(VEGF),Bcl2,Bcl-xL and Proliferating cell nuclear antigen in pancreatic tissue was significantly upregulated in MSC-treated mice compared with control mice.Additionally,protein arrays indicated that MenSC and UcMSC transplantation significantly downregulated the serum levels of interferonγand tumor necrosis factorαand upregulated the serum levels of interleukin-6 and VEGF in the model mice.Additionally,histological and immunohistochemical analyses revealed that MSC transplantation systematically improved the morphologies and functions of the liver,kidney,and spleen in T1D model mice.CONCLUSION MenSC transplantation significantly improves the symptoms in T1D model mice and exerts protective effects on their main organs.Moreover,MSC-mediated angiogenesis,antiapoptotic effects and immunomodulation likely contribute to the above improvements.Thus,MenSC are expected to become promising seeding cells for clinical diabetes treatment due to their advantages mentioned above.
文摘Objective To investigate the regulatory effect of multifactor on the matrix metalloproteinases-9 （MMP-9） and the tissue inhibitor of metalloproteinase-1 （TIMP-1） in endometrial stromal cells. Methods The endometrial stromal cells separated from the proliferative endometrial tissues were incubated with medium alone, 17-β estradiol （E2,10^-8 mol/L）, medroxyprogesterone acetate （MPA, 10^-6 mol/L）, E2（10^-8 mol/L）＋MPA （10^-6 mol/L）, E2 （10^-8 mol/L）＋MPA （10^-6 mol/L）＋RU486 （10^-5 mol/L） or HB-EGF （10 ng/ml） for 48 h respectively. The expressions of MMP-9 and TIMP-1 were detected by in situ hybridization, immunocytochemistry, reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting. Results Compared with control group [mRNA, 0. 729 ± 0. 090 （MMP-9） and 1.056± 0.154 （TIMP-1）; protein, 0.545 ±0.086 （MMP-9） and 0.745 ±0.154 （TIMP-1）], expressions of MMP-9 and TIMP-1 in E2 alone, progestin alone or E2 combined with progestin group were respectively：mRNA, 0.413 ± 0.069, 0.402 ± 0.073 and 0.407 ± 0.039; 0.487 ± 0.093, 0.503 ± 0.093 and 0.468 ± 0.075：protein, 0.294 ± 0.076, 0.331 ±0.064 and 0.265 ±0.049; 0.425 ±0.085, 0.397 ±0.065 and 0.435 ± 0.099. RU486 weakened the expression level of down-regulation, while HB-EGF elevated the level of MMP-9 and TIMP-1 after 48 h treatment （mRNA, 0.955 ± 0.068 and 1.396 ± 0.238; protein, 0. 780 ± 0.109 and 0.985 ± 0.165）. Conclusions 1） Both E2 and progestin can down-regulate the expressions of MMP-9 and TIMP-1 in endometrial stromal cells, but RU486 can inhibit the effect. 2） HB-EGF can elevate the level of MMP-9 and TIMP-1. 3） E2, progestin and HB-EGF have effect on the ratio of MM-P/TIMP-1.
文摘Objective： To observe the effect of the artesunate （ART） on cellular proliferation in vitro, to search for the possible anti-tumor mechanism of ART on endometrial carcinoma at the molecular level and to provide the experimental and theoretical foundations for the clinical applications of ART. Methods： The cell proliferation was observed by microscope; MTT was used to examine the effects of ART on proliferation of HEC-1B cells, and flow cytometric analysis was used to detect cell cycle and apoptosis. The human endometrial carcinoma HEC-1B cells were conventionally cultured; ART was administered with a concentration of 40 μg/ml before the total RNA were extracted, mRNA expression of Survivin, Caspase-3, N-Cadherin, E-Cadherin, Fibronectinl and Cox-2 were detected using RT-PCR. Results： ART reduced proliferation in human endometrial carcinoma cell line HEC-1B in a dose- and time-dependent effect. The cells of G0/G1 stage were significantly increased （P〈0.05）, but the cells of G2/M stages were significantly decreased （P〈0.05）, so it has shown that the cell cycle was probably blocked in G0/G1 stage. After intervention with ART at 20 and 80 μg/ml for 48 h, cellular apoptosis rate respectively was （36.42±0.77）% and （11.77±0.58）%, and the difference was statistically significant compared with the control （[6.64±0.191%, P〈0.01）. The expression of Cox-2 mRNA in the ART group was lower than those of control group, yet the expression of Caspase-3 and E-Cadherin mRNA in the ART group was higher than those of control group. Conclusion： ART can inhibit HEC-1B cell growth and proliferation in a dose- and time-dependent manner. Furthermore, ART can induce apoptosis in a dose-dependent manner. ART is able to downregulate Cox-2 mRNA expression and to upregulate E-Cadherin and Caspase-3 mRNA expression. So we can conclude that ART could induce the endometrial carcinoma HEC-1B cell apoptosis and inhibit tumor cell proliferation.
基金supported by Chinese National Natural Science Foundation (No: 30472227, 30572405)
文摘Objective To investigate differentially expressed genes related to inflammation in endometrial cells induced by Lipopolysaccharide（LPS）. Methods Normal endometrium in the proliferative phase of specimen from 3 cases for the experiment was collected. The LPS group were treated with 50 μg/ml LPS. Total RNA was extracted using Trizol reagent from cells. RNA quality was assessed by determining the OD260/280 ratio by agarose gel electrophoresis, the chip was scanned by laser scanner. The acquired was analyzed. Results A total of differentially expressed genes were found, these genes were relative to many aspects. Among them, the expression of genes involved in inflammation were up-regulated by LPS, such as overexpression of IL-1β, 8, etc. Conclusion The results indicates that inflammation-related genes may be one of the mechanisms of abnormal uterine bleeding by LPS-induced.
基金Supported by the National Natural Science Foundation of China(30801238)
文摘The purpose of this study was to explore the change of telomerase in passage from human endometrial stromal stem cells isolated from human endometrium.Telomerase activity of cultured endometrial stromal cells was assessed at mRNA and protein levels using RT-PCR and immunohistochemistry technique.Telomerase mRNA and protein levels were higher at early passages,and then had a gradually decreased trend of immunoreactive intensity and gradually weakened positive cells with progressive passage in endometrial stromal stem cells.These results suggest that telomerase is contributed to the insenecence of endometrial stem cell.
基金Supported by the Innovation Foundation For Postgraduate of Guangxi University(2008105930905D001) the Tackle Key Program in Science and Technology of Science and Technology Bureau of Guangxi Province(0815008-2-4)~~
文摘[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [Method] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated cells. [Result] The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purity was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [Conclusion] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purity.
文摘Objective To explore the effect of genistein on proliferation of human endometrial endothelial cells （HEECs） and glandular epithelium. Methods In vitro HEECs and human endometrial cancer-1B cell （HEC-1B ） were cultured with 0, 1, 10, 50, 100, and 200 μmol/L of genistein alone or indicated concentrations of genistein combined with 0.2 or 1 nmol/L 17β- estradiol （ 17β-E2）. Cell proliferation was determined by [ 3H ] -thymidine incorporation and cell cycle was measured by flow cytometry. Results After 96 hours of treatment, genistein inhibited the proliferation of HEECs in a dose-dependent manner. The stimulation index reduced from 100% （ without genistein treatment ） to about 1% （ 200 μmol/L genistein ）. HEECs were arrested at G1/0 and G2/M phase when treated with genistein for 96 hours. When the concentration of genistein was 200 μmol/L, the percentages of HEECs at G1/0, G2/M, and S phase were 96. 0%, 2.1%, and 1.9%, respectively. However, when HEECs were treated without genistein, the percentages of HEECs at G1/0, G2/M, and S phase were 76. 7%, 8.5%, and 14. 7%, respectively. 1713-E2 could not influence the effects of genistein on the proliferation of HEECs. Meanwhile, genistein could suppress the proliferation of HEC-1B. If the stimulation index of HEC-IB was defined as 100% when HEC-1B was treated with different doses of 1713-E2 （without genistein）, it was 67%, 19%, as well as 32% when cell was supplemented with 200 μmol/L genistein combined with 0, 0. 2, or 1 nmol/L 17β-E2, respectively. Conclusion Genistein at the concentration of 200 μmol/L can sufficiently inhibit the proliferation of HEECs and endometrial glandular epithelium simultaneously in vitro.
文摘Primary small cell carcinoma（SCC） is a group of aggressive neoplasms that mainly arise from the lung and digestive tract. Endometrial small cell carcinoma（ESCC） is extremely rare. To our knowledge, less than 90 cases have been reported, and most of these reports were dedicated to describing the clinicopathologic or immunochemical features of ESCC. Herein, we present a new case of ESCC involving a 51-year-old woman and mainly focus on the magnetic resonance imaging（MRI） and positron emission tomography/computed tomography（PET/CT） findings. MRI showed that the uterus was significantly enlarged（11.6 cm × 11.1 cm × 14.4 cm）, and a giant irregular mass（7.5 cm × 8.4 cm × 8.5 cm） was observed in the uterine cavity. The lesion demonstrated an extremely low apparent diffusion coefficient（ADC） value [（0.553±0.088）×10^–3 mm^2/s] and a high FDG uptake value（22.7）. Multiple metastatic lymph nodes（LNs） were identified at different positions, with diameters ranging from 0.3 to 2.8 cm and a maximum standardized uptake value（SUV max） ranging from 6.9 to 19.3.
文摘BACKGROUND Cervical squamous cell carcinoma(SCC)is the most common type of cervical carcinoma and is generally derived from a precancerous stage called cervical high-grade squamous intraepithelial lesion(HSIL).Usually,the cancer metastasizes through lymphatic or hematogenous dissemination,but rarely spreads upward into the uterus.Here,we report a case of cervical HSIL extending into the endometrium and finally progressing to SCC in the uterine cavity.CASE SUMMARY A 57-year-old postmenopausal woman visited our department and requested a routine cervical check-up.Four years ago,she had undergone a cervical loop electrosurgical excision procedure because of HSIL found during the gynecological examination,and she had not been checked again since.This time,a relapse of the cervical HSIL was diagnosed along with uterine pyometra and endometrial polyps.After 2 wk of antibiotic treatment,a laparoscopic hysterectomy was performed,and the final pathological examination revealed that the cervical HSIL had spread directly upward into the uterine cavity,gradually developing into cervical SCC in the endometrium.CONCLUSION Cervical HSIL/SCC can directly spread upward into the uterus with the most common symptoms of pyometra and cervical stenosis.More attention should be given to the early detection and prevention of this disease.
文摘s To study the influence of insulin on IGF Ⅰ and IGFBP Ⅰ secretion of the hum an endometrial stromal cells Methods Late proliferative phase endometrial stromal cells were isolated from endometriu m tissues and then cultured for 24 h in Hams F 12 only as a control and in Hams F 12 with different concentrations of estradiol (E2) and insulin (INS) as trea ted groups Simultaneously, the endometrial stromal cells from late secretory phase endometrium were cultured for 24 h in Hams F 12 only as a control and in Hams F 12 supplemented with different concentrations of progesterone (P) and i nsulin as treated groups After 24 h of culturing, the mediums were collected f or either IGF Ⅰ or IGFBP Ⅰ assays Result The concentrations of IGF Ⅰ in medium from cultured endometrial stromal cells i n the proliferative phase were 0 78±0 47 ng/ml in the hormone free control g roup; 1 44±0 59 ng/ml and 1 39± 0 33 ng/ml in 100 pg/ml E2 group and 20 μU /ml INS group, which was higher than that of the control group ( P <0 05 and P <0 01, respectively) The IGF Ⅰ concentration in the 100 μU/ml INS group was 2 03±0 53 ng/ml, which was higher than that of the 20 μU/ml INS group ( P <0 01) Levels of IGF Ⅰ in the 100 pg/ml E2 plus 20 μU/ml INS gro up was 2 18±0 36 ng/ml, which was significantly higher than that of the 20 μU/ml INS and 100 pg/ml E2 group ( P <0 01), but lower than that of the 100 pg/ml E2 p lus 100 μU/ml INS group (3 42±0 75 ng/ml), P <0 01 The concentration o f IGFBP Ⅰ in medium from cultured endometrial stromal cells in the secretory ph a se was 2 50±1 39 ng/ml in the hormone free control group and 5 44±2 09 ng /ml in the 10 pg/ml P group, which was significantly higher than that of the con trol ( P <0 01) IGFBP Ⅰ concentration in 20 μU/ml INS group was 0 1 6±0 58 ng/ml, which was lower compared with control, but higher compared with the 100 μU/ml INS group ( P <0 01) The level of IGFBP Ⅰ in the 10 ng/ml P plu s 20 μU/ml INS group was 2 10±1 17 ng/ml, lower compared with the 10 ng/ml P gr oup, but higher compared with the 10 pg/ml P plus 100 μU/ml INS group, P <0 01 Conclusions Insulin can stimulate basal (without hormone) and E2 stimulated IGF Ⅰ secreti on in cultured stromal cells from human late proliferative endometrium in a dose dependent manner Insulin can suppress basal (without hormone) and P stimula ted IGFBP Ⅰ secretions in cultured stromal cells from human secretory endomet rium in a dose dependent manner
基金supported by the National Key Technology R&D Program of China(No.2013BAD10B04)the Importation and Development of High-Caliber Talents Project of Beijing Municipal Institutions(No.CIT&TCD20130324),China
文摘Objective：Bovine endometritis is one of the most common reproductive disorders in cattle.The aim of this study was to investigate the anti-inflammation potential of punicalagin in lipopolysaccharide（LPS）-induced bovine endometrial epithelial cells（bE ECs）and to uncover the underlying mechanisms.Methods：bE ECs were stimulated with different concentrations（1,10,30,50,and 100μg/ml）of LPS for 3,6,9,12,and 18 h.MTT assay was used to assess cell viability and to identify the conditions for inflammatory injury and effective concentrations of punicalagin.Quantitative real-time polymerase chain reaction（q RT-PCR）was used to assess gene expression of pro-inflammatory cytokines.Western blotting was used to assess levels of inflammation-related proteins.Results：Treatment of b EECs with 30μg/ml LPS for 12 h induced cell injury and reduced cell viability.Punicalagin（5,10,or 20μg/ml）pretreatment significantly decreased LPS-induced productions of interleukin（IL）-1β,IL-6,IL-8,and tumor necrosis factor-α（TNF-α）in bE ECs.Molecular research showed that punicalagin inhibited the activation of the upstream mediator nuclear factor-κB（NF-κB）by suppressing the production of inhibitorκBα（IκBα）and phosphorylation of p65.Results also indicated that punicalagin can suppress the phosphorylation of mitogen-activated protein kinases（MAPKs）including p38,c-Jun N-terminal kinase（JNK）,and extracellular signal-regulated kinase（ERK）.Conclusions：Punicalagin may attenuate LPS-induced inflammatory injury and provide a potential option for the treatment of dairy cows with Escherichia coli endometritis.
文摘Extracellular vesicles from highly metastatic tumor cells have been shown to mediate epithelial-mesenchymal transition(EMT)-related events in recipient cells.In endometrial epithelial cells,EMT processes are known to be involved in the development of adenomyosis.We aimed to investigate whether adenomyosis-derived extracellular vesicles(AMEVs)are able to induce an EMT process in endometrial epithelial cells.In this study,AMEVs were isolated from patients with adenomyosis and characterized by transmission electron microscopy,Western blot,and nanoparticle tracking.Primary endometrial epithelial cells(EECs)were derived from normal endometrium tissues from patients with leiomyoma and co-cultured with AMEVs in vitro.AMEV uptake was examined by fluorescence confocal microscopy.The invasion of EECs was confirmed by Transwell assay.Immunohistochemistry,Western blot,and qRT-PCR were performed on EECs to illustrate the expression levels of cytokeratin 19,E-cadherin,vimentin,and zinc finger E-boxbinding homeobox 1(ZEB1).The results indicated that the cellular fluorescence intensity gradually increased after 48 h of co-culture,but decreased after 72 h.After co-culturing with AMEVs for 72 h,EECs expressed significantly lower levels of cytokeratin 19 and E-cadherin,and significantly higher levels of vimentin and ZEB1.Together these results demonstrated that AMEVs induce an EMT process and enhance the invasion of EECs.These changes may contribute to the pathogenesis and progression of adenomyosis.
基金funded by grants from the National Natural Science Foundation of China(No.81671463)the Key Research and Development Plan of Shaanxi Province(No.2017ZDCXL-SF-02-03)。
文摘Objective:This study aimed to investigate the differentiation of human adipose-derived stem cells(hASCs)into endometrial epithelial cells(EECs)under certain induction conditions and to a further step provide a promising approach for ASCs in clinical practice to the treatment of severe intrauterine adhesion.Methods:Four groups of hASCs were separately cultured as follows:in Group 1,hASCs were cultured in a control medium(5%fetal bovine serum[FBS]+α-minimum Eagle’s medium[α-MEM]);in Group 2,hASCs were cultured in an induction medium(5%FBS+α-MEM+[1×10-7 mol/L 17β-estradiol]+10 ng/mL transforming growth factorβ1[TGF-β1]+10 ng/mL epidermal growth factor[EGF]+10 ng/mL platelet-derived growth factor BB[PDGF-BB]);in Group 3,hASCs and human endometrium cells(hEMCs)were cocultured in the control medium;and in Group 4,hASCs and hEMCs were cocultured in the induction medium.Results:When cocultured with hEMCs,the morphology of hASCs became similar with EECs,and the addition of factors such as EGF,TGFβ,PDGF-BB,and 17β-estradiol promoted differentiation.This study,for the first time,demonstrated estrogen receptor(ER)αand ERβexpression in hASCs and preliminarily explored changes in ERα,ERβ,β-catenin,and H19 mRNA expression during hASC differentiation.Furthermore,we concluded that H19 mRNA expression was negatively correlated with differentiation,which is seemingly related to the estrogen signaling pathway.Conclusions:hASCs revealed the potential for differentiating to EECs when cocultured with hEMCs.
基金supported by the RGC General Research Fund(17120720 to K.F.L.)the Professor PC Ho Research and Development Fund in Reproductive Medicine from the Department of Obstetrics and Gynecology,University of Hong Kong and Health and Medical Research Fund,Hong Kong(L.H.J.)a Conference and Research Committee grant from the University of Hong Kong(K.F.L.).
文摘Endometrial stromal cell decidualization is a crucial step in endometrial remodeling during pregnancy.Decidualization is controlled by orchestrated ovarian hormones,followed by the activation of various downstream signaling pathways.Accumulating evidence has shown multiple functions of decidualized endometrial stromal cells during embryo implantation,including tissue remodeling,antioxidative stress,angiogenesis,and immune tolerance.The distinct secretomes of decidualized stromal cells also reveal their intensive interactions with epithelial,endothelial,and immune cells.However,aberrant decidualization leads to pregnancy failures,such as recurrent pregnancy loss and repeated implantation failure.This review aimed to provide an overview of the molecular mechanisms underlying the divergent functions of decidualized endometrial stromal cells and their potential clinical applications.Moreover,the use of single-cell RNA sequencing data further enhances our understanding of these biological processes.This review discusses decidualization-related signaling pathways that serve as potential therapeutic targets for treating implantation failure in in vitro fertilization and provides novel approaches to investigate the underlying causes of female infertility.
文摘Numerous studies have shown aberrant immune cell function in endometriosis,including T cells,B cells,natural killer cells,and macrophages(MΦ).These alterations are thought to be induced by various mechanisms that promote the disease.Regulatory T cells(Tregs)may account for a decreased ability of newly recruited leukocytes to initiate effective immune responses against viable endometrial fragments,permitting their survival.Tregs differentiate during the development of endometriosis,which confer immunosuppression or play other roles in disease progression.In this review,we provide an overview of the regulation and roles of Tregs in endometriosis.These data provide further scientific evidence for the altered immune response in endometriosis,which could be a potential target in the treatment of endometriosis.This review could create new diagnostic strategies and effective immune-targeted therapies for this highly prevalent disease.Recent progress in the field indicates that these goals may be achieved in the future.
