Celecoxib attenuates interleukin 33-induced expression of vascular cell adhesion molecule-1 in human ovarian endometriotic stromal cells.

OBJECTIVE: Endometriosis is an estrogen-dependent chronic inflammatory disease in women of reproductive age. A review of the literature revealed that cytokines and inflammatory factors are associated with endometriosis-associated infertility. Interleukin 33 (IL-33) is a strong inducer of other pro-inflammatory cytokines. Vascular cell adhesion molecule-1 (VCAM-1) plays a central role in recruiting inflammatory cells, whose expression facilitates leukocyte adhesion and is rapidly induced by pro-inflammatory cytokines. Many studies have indicated that VCAM-1 expression is high in endometriosis; however, whether the expression of VCAM-1 is related to IL-33 is unclear. MATERIALS AND METHODS: Human ovarian endometriotic stromal cells (hOVEN-SCs) were treated with IL-33 to enable investigation of cell characterization, gene and protein expression, and signal pathways. Proliferation potential was measured using an MTT assay. Gene expression was analyzed using reverse transcription-polymerase chain reaction. Protein expression assay was performed using western blot analysis. RESULTS: This study investigated the effects of IL-33 on VCAM-1 and COX-2 expression in hOVEN-SCs. First, the results revealed that the IL-33/ST2/mitogen-activated protein kinase (MAPK) signaling pathway could increase the expression of VCAM-1 and COX-2 in hOVEN-SCs. Second, we discovered that COX-2 expression was essential for IL-33-induced VCAM-1 expression because the effects could be negated through NS398, a selective COX-2 inhibitor. Finally, treatment of IL-33-treated hOVEN-SCs with celecoxib significantly and dose-responsively decreased VCAM-1 expression. CONCLUSION: Taken together, these results indicate that IL-33 can upregulate VCAM-1 expression in hOVEN-SCs through the IL-33/ST2/MAPK/COX-2 signaling pathway and thereby contribute to endometriosis.

Segmentation of In vitro Fertilization with High-intensity Focused Ultrasound in Repeated Implantation Failure with Adenomyosis.

Adenomyosis is a complex issue in reproductive-age women not only on worsening of quality of life due to severe dysmenorrhea or heavy menstrual bleeding but also on the impact of infertility. A 39-year-old female, gravida 0 para 0, with a history of bilateral ovarian endometrioma post laparoscopic surgery presented to our hospital due to suspected deep infiltrative endometriosis (DIE), adenomyosis, and repeated implantation failure. Initially, gonadotropin-releasing hormone analog treatment for DIE with progestin-primed ovarian stimulation protocol was arranged. Four D5 blastocysts were obtained and freezed. Two frozen embryo transfer were performed after ultrasound-guided high-intensity focused ultrasound (USgHIFU) treatment of adenomyosis. She later had a dichorionic diamniotic twin pregnancy, and two healthy newborns were delivered by Cesarean section at gestational age of 35 weeks due to antepartum hemorrhage with placenta previa and preeclampsia. In conclusion, USgHIFU can be a potential treatment option in segmented in vitro fertilization in future.

Oct-4 induces cisplatin resistance and tumor stem cell-like properties in endometrial carcinoma cells.

OBJECTIVE: Research has suggested that tumor-initiating tumor stem cells are derived from normal stem cells and that tumor cells undergo progressive de-differentiation to achieve a stem cell-like state. Tumor stem cells are characterized by high proliferation ability, high plasticity, expression of multi-drug resistance proteins, and the ability to seed new tumors. Octamer-binding transcription factor 4 (Oct-4) and its activation targets are overexpressed in the tumor stem cells of various types of tumors, and this expression is associated with the pathogenesis, development, and poor prognosis of tumors. The primary objective of this study was to test if a stably transfected with Oct-4 gene cell line, RL95-2/Oct-4, has the characteristics of tumor stem cells. MATERIALS AND METHODS: Human endometrial carcinoma cells (RL95-2) were transfected with a plasmid carrying genes for Oct-4 and green fluorescent protein (GFP). The stably transfected cells, RL95-2/Oct-4, were selected using G418 and observed to express the GFP reporter gene under the control of the Oct-4 promoter. GFP expression levels of RL95-2/Oct-4 cells were measured using flow cytometry. The proliferation potential of cells was determined according to cumulative population doubling and colony-formation efficiency. Gene expression was analyzed using reverse transcription-polymerase chain reaction. RESULTS: RL95-2/Oct-4 cells not only exhibited increased expression of the three most important stem cell genes, Oct-4, Nanog, and Sox2, but also had increased expression of the endometrial tumor stem cell genes CD133 and ALDH1. Furthermore, enhanced expression of these genes in the RL95-2/Oct-4 cells was associated with higher colony-forming ability and growth rate than in parental RL95-2 cells. We also observed that cisplatin induced less cell death in RL95-2/Oct-4 cells than in RL95-2 cells, indicating that RL95-2/Oct-4 cells were more resistant to chemotherapeutic agents. CONCLUSION: The study findings contribute to investigate the effects of Oct-4 on tumor stem cell origins.

Complex rearrangements of Y chromosome suggest RPS4Y1 as lymphedema candidate gene.

OBJECTIVE: Cystic hygromas are frequently encountered in fetus with Turner syndrome (TS). Nevertheless, identification of genetic loci responsible for the cystic hygroma has been problematic. Here, we tried to elucidate the candidate gene for cystic hygroma through a rare case of complex Y chromosomal rearrangements involving duplication of partial Yq and monosomy of partial Yp. CASE REPORT: A 30-year-old woman, gravida 1 para 0, was diagnosed with fetal cystic hygroma at 12 weeks of gestation. The genetic analysis of the product of conception revealed complex rearrangement of Y chromosome: microdeletion in Yp11.2p11.31 and microduplicatin in Yq11.223q11.23. The deleted region spans about 6.25 Mb and includes 76 genes, including SRY. The duplicated region spans about 4.76 Mb and includes 145 genes. CONCLUSION: From this rare case with non-mosaic complex Y-chromosome rearrangements, we could narrow down Turner stigmata critical region to Yp11.2～p11.3. We also propose RPS4Y1 as lymphedema candidate gene.

Pretreatment bowel manipulation during ultrasound-guided high-intensity focused ultrasound therapy for posterior wall uterine masses.

OBJECTIVE: High-intensity focused ultrasound (HIFU) therapy is a noninvasive alternative to conventional abdominal surgery in obstetrics and gynecology. The aim of this study is to evaluate the reduction of pain intensity with bowel manipulation before ultrasound-guided HIFU treatment in women with posterior wall uterine fibroids and/or adenomyosis. MATERIALS AND METHODS: This is a multicenter retrospective observational study. Data from all patients who underwent HIFU therapy at three HIFU clinics (Sichuan Maternal and Child Health Hospital, Xiangya Hospital of Central South University, and Kuo General Hospital) between January 2019 and December 2019 were analyzed. We compared pain intensity with and without bowel manipulation during the HIFU treatment and evaluated tolerability without intravenous sedation. The presence of discomfort or pain during the HIFU procedure was evaluated using the visual analog scale (VAS). RESULTS: A total of 86 women were included in this study. All women underwent HIFU therapy with the PRO-2008 system in the supine position for posterior wall uterine fibroids and/or adenomyosis. Thirty-seven women received pretreatment anal catheterization with a condom and 49 women were not subjected to bowel manipulation. All patients received pretreatment condom-catheter device were well tolerated during the procedure of bowel manipulation. During the HIFU procedure, the women who had received bowel manipulation experienced lower pain intensity, especially less sacrococcygeal pain (VAS score 1.56 ± 1.46 vs 2.89 ± 1.61), target region pain (1.54 ± 1.30 vs 2.53 ± 1.29), and radiating pain (0.13 ± 0.34 vs 0.41 ± 0.54), compared with the women without bowel manipulation. CONCLUSION: Bowel manipulation with anal catheterization before HIFU therapy for posterior wall uterine masses can be safely performed and is effective as a low risk intervention to aid in reducing potential HIFU complications related to nerve involvement.

A survey of career-decision factors for obstetrics and gynecology residents in Taiwan.

OBJECTIVE: The recruitment of obstetrics and gynecology residents has been challenging in Taiwan since 2000. There are a lots factors influencing applying, including career interest and prospects, lifestyle, salary, litigation, stress level, time demands of specialty work and gender consideration. The purpose of this study was to evaluate the factors influencing the career choice of current OB/GYN residents and identify the important factors influencing recruitment. MATERIALS AND METHODS: A cross-sectional questionnaire of career-decision factors was sent to all 280 residents during December, 2019. Total 16 factors were rated on a 5-point Likert and were ranking by their average score on the scale. Gender of residents was analyzed for their association with the scores by a two-sample test and two-tailed t test. RESULTS: 91 (32.5%) residents responded and 61 were female and 30 were male. The top influencing factor was being interested in clinical expertise with an average score of 4.32. The second factor was having a great sense of accomplishment in saving people with an average score of 3.82, followed by it being easier to become a medical practitioner at 3.77. The lowest scoring factor was fewer hours of work, with an average score of 1.77 and a safer working environment with a score of 2.33 was the second lowest factor. Female residents agreed that it was easier to get specialist certification with a score of 3.0. This score was significantly higher than male doctors at 2.63 with p value of 0.017. CONCLUSION: Career interest and prospects were key influencing factors for applying OB/GYN residents. Minimizing the influence of detracting factors like heavy workload and medical litigation was also effective. Enrolling new residents is key to maintaining adequate staffing in specialties in healthcare. Establishing a safer work environment and determining the optimal workload will be the next reforms in the future.

Interleukin-33 promotes invasiveness of human ovarian endometriotic stromal cells through the ST2/MAPK/MMP-9 pathway activated by 17ß-estradiol.

OBJECTIVE: Endometriosis is an estrogen-dependent, benign, and chronic gynecological disorder occurring in women of reproductive age. Although the pathogenesis of endometriosis is poorly understood, implantation theory indicates that viable endometrial cells shed from the endometrium into the pelvic peritoneum or ovaries, possibly through retrograde menstruation, and then reattach, invade, and damage other tissues. Interleukin (IL)-33, a new member of the IL-1 superfamily, is mainly upregulated by stromal cells following proinflammatory stimulation. Matrix metalloproteinases (MMPs) are involved in the degradation and reconstruction of the extracellular matrix. MMP-9 participates in the pathogenesis of endometriosis by promoting the invasion of endometriotic cells. This study investigated the effect of IL-33 on the cell invasion ability of and MMP-9 expression in human stromal cells derived from ovarian endometrioma (hOVEN-SCs). MATERIALS AND METHODS: We isolated hOVEN-SCs from human ovarian endometrioma. Gene expression was analyzed using the Illumina Human WG-6 v2 Expression BeadChips microarray platform and through reverse transcription-polymerase chain reaction. Cell migration and invasion were examined by performing the transwell chamber assay. RESULTS: We found that 17ß-estradiol could increase the expression of IL-33 and ST2 through the estrogen receptor pathway in hOVEN-SCs. Moreover, IL-33 upregulated MMP-9 expression in and enhanced the invasion ability of hOVEN-SCs through the ST2/MAPK signaling pathway. Our results showed that MMP-9 expression was essential for IL-33-induced cell invasion. CONCLUSION: Our main finding is that 17ß-estradiol could increase IL-33 expression through the estrogen receptor pathway and activate MMP-9 expression in and invasion ability of hOVEN-SCs through the IL-33/ST2/MAPK signaling pathway. The results of this study and further related studies may provide new strategies for the prevention and treatment of endometriosis.

Downregulation of gap junctional intercellular communication and connexin 43 expression by bisphenol A in human granulosa cells.

Gap junctional intercellular communication (GJIC) is the transfer of ions, metabolites, and second messengers between neighboring cells through intercellular junctions. Connexin 43 (Cx43) was found to be the type of gap junction protein responsible for human granulosa cells (GCs) and oocyte communication, which is required for folliculogenesis and oocyte maturation. Bisphenol A (BPA), an estrogenic-like endocrine-disrupting chemical, is one of the most widely produced chemicals around the world. There are reports that the chemical might cause endometrial tumorigenesis and several female reproductive disorders. This study demonstrated that cell culture medium, containing antioxidants (N-acetyl-l-cysteine and l-ascorbic acid-2-phosphate), was able to enhance the survival and self-renewal of GCs. In addition, we found that BPA at environmentally relevant concentration (10-7  M) reduced Cx43 expression and GJIC in GCs through estrogen receptor and mitogen-activated protein kinase pathways. The results of this study not only reveal the reproductive toxicity of BPA but also provide possible mechanisms by which BPA inhibited GJIC in GCs.

Modulation of tumor stem cell characteristics by 17ß-estradiol in human mesenchymal stem cells derived from ovarian endometrioma.

OBJECTIVE: Ovarian endometrioma is a cyst composed of endometrial tissue and is present in 20%-40% of patients with endometriosis. Endometriosis is an estrogen-dependent benign and chronic gynecological disease that affects women of reproductive age. Studies have reported that tumor stem cells can be isolated from numerous tumor types. Emerging evidence has indicated that tumor stem cells may be responsible for the development of endometriosis and endometrial tumors. The present study investigated the effects of 17ß-estradiol on levels of expression of stem cell markers and cell growth of human mesenchymal stem cells derived from ovarian endometrioma (hOVEN-MSCs). MATERIALS AND METHODS: hOVEN-MSCs were isolated from human ovarian endometrioma. The proliferation potential of hOVEN-MSCs was measured by the cumulative population doubling and colony-formation efficiency. The gene expression of the hOVEN-MSCs was examined by the reverse transcription-polymerase chain reaction analysis. Protein expression assays were performed using flow cytometry and western blot analysis. RESULTS: This study demonstrated that hOVEN-MSCs can be isolated from ovarian endometrioma and that 17ß-estradiol was capable of increasing colony-forming efficiency and cell proliferation of these cells. In addition, we found that 17ß-estradiol not only increased the expression of the stem cell marker OCT-4, but also increased the expression of endometrial tumor stem cell markers CD133 and ALDH1 in hOVEN-MSCs. CONCLUSION: The above results indicate an important role of 17ß-estradiol in cell growth of hOVEN-MSCs concomitant with enhanced expression of stem cell markers. This effect of 17ß-estradiol related to stem cell marker expression, if confirmed by further in vitro, in vivo studies, may be useful for developing new strategies for prevention and treatment of endometriosis and endometrioma.

Work hours of employed obstetricians in Taiwan.

OBJECTIVE: Obstetricians have longer work hours and work hour restriction of employed physicians will be implemented thereafter. This study was to survey the impact of work hour restriction and determine the weekly work hours of obstetricians. MATERIALS AND METHODS: Since December 1, 2017, to January 31, 2018, a questionnaire about reasonable workload investigation, impact of work hour restriction and weekly work diary was mailed to 196 employed obstetricians who delivered more than 20 newborns per months. The work types and coefficient of correlation with work hours was evaluated using the Pearson correlation coefficient (r). RESULTS: 72 returned the questionnaires, 21 (31.4%) believed that 40-49 h were reasonable, while 22 (32.8%), 13 (19.4%), 7 (10.4%), and 4 (6%) thought that 50-59, 60-69, 70-79, and >80 h were reasonable. 66 completed their weekly work diary. The average weekly work hours were 80.14 ± 6.85. If the time of awaiting delivery was calculated into total work hours, then the average work hours were 116.96 ± 12.41. The coefficient of correlation between on-duty and weekly work hours was 0.7. The average work hours of physicians with on-duty work were 92.08 ± 8.7, which were significantly higher than the work hours of those without on-duty works (63.95 ± 7.79). CONCLUSION: The weekly work hours of employed obstetricians is higher than their expected reasonable work hours. Implementation of work hour restriction will have a significant impact on medical care. It is important to determine a reasonable work hour restriction and modify the patient care model to reduce workload.

Enhancing the Microparticle Deposition Stability and Homogeneity on Planer for Synthesis of Self-Assembly Monolayer.

The deposition stability and homogeneity of microparticles improved with mask, lengthened nozzle and flow rate adjustment. The microparticles can be used to encapsulate monomers, before the monomers in the microparticles can be deposited onto a substrate for nanoscale self-assembly. For the uniformity of the synthesized nanofilm, the homogeneity of the deposited microparticles becomes an important issue. Based on the ANSYS simulation results, the effects of secondary flow were minimized with a lengthened nozzle. The ANSYS simulation was also used to investigate the ring-vortex generation and why the ring vortex can be eliminated by adding a mask with an aperture between the nozzle and deposition substrate. The experimental results also showed that particle deposition with a lengthened nozzle was more stable, while adding the mask stabilized deposition and diminished the ring-vortex contamination. The effects of flow rate and pressure were also investigated. Hence, the deposition stability and homogeneity of microparticles was improved.

Pueraria mirifica inhibits 17ß-estradiol-induced cell proliferation of human endometrial mesenchymal stem cells.

OBJECTIVE: The notion that the human endometrium may contain a population of stem cells has recently been proposed. The mesenchymal stem cells (MSCs) in the endometrium are believed to be responsible for the remarkable regenerative ability of endometrial cells. Estrogens influence the physiological and pathological processes of several hormone-dependent tissues, such as the endometrium. Pueraria mirifica (PM) is a herbal plant that contains several phytoestrogens, including isoflavones, lignans, and coumestans, and is known to exert an estrogenic effect on animal models. The present study investigated the effects of PM on the proliferation of human endometrial MSCs (hEN-MSCs). MATERIALS AND METHODS: The hEN-MSCs were isolated from human endometrial tissue. The surface markers of these hEN-MSCs were identified through reverse transcription-polymerase chain reaction analysis. The proliferation potential of hEN-MSCs was measured through a cell proliferation assay. Multilineage differentiation ability was confirmed through Oil red O and von Kossa staining. RESULTS: This study demonstrated that 17ß-estradiol-responsive MSCs with Oct-4, CD90, and CD105 gene expression can be derived from the human endometrium and that PM exerts biological effects on hEN-MSCs, specifically, enhanced cell growth rate, through the estrogen receptor. Furthermore, PM at 1500 and 2000 µg/mL significantly increased cell proliferation compared with the vehicle control, and PM concentration at 1000 µg/mL significantly inhibited the enhanced cell growth rate induced by 17ß-estradiol in hEN-MSCs. CONCLUSION: This study provides new insights into the possible biological effects of PM on the proliferation of hEN-MSCs.

Lifespan Extension and Sustained Expression of Stem Cell Phenotype of Human Breast Epithelial Stem Cells in a Medium with Antioxidants.

We have previously reported the isolation and culture of a human breast epithelial cell type with stem cell characteristics (Type I HBEC) from reduction mammoplasty using the MSU-1 medium. Subsequently, we have developed several different normal human adult stem cell types from different tissues using the K-NAC medium. In this study, we determined whether this low calcium K-NAC medium with antioxidants (N-acetyl-L-cysteine and L-ascorbic acid-2-phosphate) is a better medium to grow human breast epithelial cells. The results clearly show that the K-NAC medium is a superior medium for prolonged growth (cumulative population doubling levels ranged from 30 to 40) of normal breast epithelial cells that expressed stem cell phenotypes. The characteristics of these mammary stem cells include deficiency in gap junctional intercellular communication, expression of Oct-4, and the ability to differentiate into basal epithelial cells and to form organoid showing mammary ductal and terminal end bud-like structures. Thus, this new method of growing Type I HBECs will be very useful in future studies of mammary development, breast carcinogenesis, chemoprevention, and cancer therapy.

Bisphenol A-induced epithelial to mesenchymal transition is mediated by cyclooxygenase-2 up-regulation in human endometrial carcinoma cells.

Many studies have highlighted the correlation between the increase of bisphenol A (BPA) level in the environment and the incidence of tumor in humans. In human carcinogenesis, the overexpression of cyclooxygenase-2 (COX-2) and epithelial-mesenchymal transition (EMT) are closely related with tumor development. In this study, human endometrial carcinoma cells line (RL95-2) was used to investigate whether BPA can induce EMT and COX-2 expression. The results show that BPA increased growth rate and colony-forming efficiency in a dose-dependent manner, induced EMT and COX-2 gene expression and promoted the migration and invasion ability of RL95-2 cells. Furthermore, our study showed that the expression of COX-2 was essential for BPA-induced cell migration and invasion. The results of this study provide new insights into the mechanism of endometrial cancer cell growth and invasion and potential therapeutic strategy.

Upregulation of Nanog and Sox-2 genes following ectopic expression of Oct-4 in amniotic fluid mesenchymal stem cells.

Octamer-binding transcription factor 4 (Oct-4), an important gene regulating stem cell pluripotency, is well-known for its ability to reprogram somatic cells in vitro, either alone or in concert with other factors. The aim of this study was to assess the effect of ectopic expression of Oct human amniotic fluid stem cells. We developed a novel method for isolation of putative human amniotic fluid-derived multipotent stem cells. These cells showing mesenchymal stem cell phenotypes (human amniotic fluid-derived mesenchymal stem cells, hAFMSCs) were transfected with a plasmid carrying genes for Oct-4 and the green fluorescent protein (GFP). The stably transfected cells, hAFMSCs-Oct4/GFP, were selected by using G418 and found to express the GFP reporter gene under the control of Oct-4 promoter. We found that hAFMSCs developed by our method possess very high self-renewal ability (about 78 cumulative population doublings) and multilineage differentiation potency. Significantly, the hAFMSCs-Oct4/GFP cells showed enhanced expression of the three major pluripotency genes Oct-4, Nanog, and Sox-2, and increased colony-forming ability and growth rate compared with the parental hAFMSCs. We demonstrated that the ectopic expression of Oct-4 gene in hAFMSCs with high self-renewal ability could upregulate Nanog and Sox-2 gene expression and enhance cell growth rate and colony-forming efficiency. Therefore, the ectopic expression of Oct-4 could be a strategy to develop pluripotency in hAFMSCs for clinical applications.

A non-mosaic isodicentric Y chromosome resulting from breakage and fusion at the Yq pseudo-autosomal region in a fetus.

The dicentric Y chromosomes are the most commonly found in the structural aberration of Y chromosome. If the dicentric chromosome has completely symmetric arms, it is considered an isodicentric chromosome. The sites of breakage and fusion at Yp and Yq are variable, but breakage and fusion at the pseudo-autosomal region has never been reported. Herein we reported identification de novo isodicentric (Yq12) in a fetus. The fusion occurred at Yq pseudo-autosomal region very close the telomere and resulted in duplication of Y chromosome. The baby was grossly normal at birth. In conclusion, isodicentric Y chromosome could result from breakage and fusion at the Yq pseudo-autosomal region.