Hung-Sheng Chen

Correlations between umbilical and maternal serum adiponectin levels and neonatal birthweights

Objective.  To measure adiponectin levels in maternal serum and umbilical cord serum at delivery, and examine whether or not there are correlations between adiponectin levels and neonatal birthweights, maternal body weights and body mass indexes.

Comparative study of human eutopic and ectopic endometrial mesenchymal stem cells and the development of an in vivo endometriotic invasion model

OBJECTIVE To elucidate the role of endometrial stem-progenitor cells in the etiology of endometriosis and to develop an animal model to study the invasion ability of endometrial cells. DESIGN Gene expression and cell function studies were designed. SETTING Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan. PATIENT(S) Human endometrial mesenchymal stem cells (MSCs) were isolated from 22 different endometrium biopsies after surgery for treatment of endometriosis. INTERVENTION(S) Endometrial MSCs developed from eutopic and ectopic endometrial tissues. MAIN OUTCOME MEASURE(S) Characterization of MSC phenotypes (i.e., differentiation induction and gene expression by flow cytometric analysis); comparative study of cell functions (i.e., cell growth, migration, and invasion assays). The invasion of implants in an animal model was examined by histologic staining. RESULT(S) We compared the characteristics of eutopic and ectopic endometrial MSCs from the same endometrial donor. Although both showed similar mesenchymal cell phenotypes, ectopic endometrial MSCs showed distinctly greater ability of cell migration and invasion. Furthermore, in an in vivo cell invasion model using cells grown in scaffold and transplantation in immune-deficient mice, the ectopic endometrial MSCs were found to form many new blood vessels and to invade surrounding tissue. CONCLUSION(S) These results indicate unique invasion and angiogenesis characteristics of ectopic endometrial MSCs that may underlie the pathogenesis of ectopic endometriosis. The animal invasion model will be useful for future characterization of endometrial MSCs.

Increased Serum Retinol-Binding Protein 4 Concentrations in Women With Gestational Diabetes Mellitus

The authors hypothesized that serum retinol-binding protein 4 (RBP4) concentrations will be higher in gestational diabetes mellitus (GDM) subjects. This study tested both women with GDM and healthy pregnant women and correlated their serum RBP4 concentrations with body mass index (BMI) and a variety of other parameters. Also, since there is no information on the relationship between RBP4 concentrations in maternal and fetal serum, this study measured these at delivery and examined whether there were correlations between the cord serum RBP4 levels and maternal serum RBP4 concentrations, neonatal birth weights, and gestational age at delivery. A total of 40 women were evaluated: 20 women with GDM and 20 healthy pregnant women to serve as control subjects. Serum RBP4 concentrations were analyzed with the use of an enzyme-linked immunosorbent assay kit. Serum RBP4 concentrations at glucose challenge test (GCT) were significantly higher in the GDM group (42.4 ± 13.8 ng/mL) than in the healthy control group (32.0 ± 8.7 ng/mL; P = .007). BMI at GCT (P = .003) and GDM/no GDM (P = .014) were significantly correlated to serum RBP4 concentrations at GCT by multiple linear regression analysis. In GDM subjects, serum RBP4 concentrations immediately after delivery were significantly lower than those at GCT (30.1 ± 11.0 ng/mL, 42.4 ± 13.8 ng/mL; P < .001), but there was no such difference in normal subjects (30.9 ± 10.0 ng/mL, 32.0 ± 8.7 ng/mL; P = .581). Cord serum RBP4 concentrations were significantly lower than maternal serum RBP4 concentrations at delivery (10.9 ± 3.8 ng/mL, 30.5 ± 10.4 ng/mL; P < .001). Only fetal birth weight (P = .049) was independently related to cord serum RBP4 concentrations at delivery by multiple linear regression analysis. This study found increased serum RBP4 concentrations at GCT in GDM subjects, and GDM was significantly correlated to serum RBP4 levels after adjustment for the effect of BMI. Lower RBP4 concentrations were found at delivery in GDM subjects. Maternal serum RBP4 concentrations were significantly higher than cord serum RBP4 concentrations, and fetal birth weights were independently correlated to cord serum RBP4 concentrations. These findings may indicate that RBP4 plays a role in the pathogenesis of GDM. However, further experiments are required to clarify this role and find a possible regimen for GDM treatment.

miRNA-199a-5p regulates VEGFA in endometrial mesenchymal stem cells and contributes to the pathogenesis of endometriosis.

It is believed that endometrial miRNAs contribute to the aetiology of endometriosis in stem cells; however, the mechanisms remain unclear. Here we collected serum samples from patients with or without endometriosis and characterized the miRNA expression profiles of these two groups. MicroRNA‐199a‐5p (miR‐199a‐5p) was dramatically down‐regulated in patients with endometriosis compared with control patients. In addition, we found that the tumour suppressor gene, SMAD4, could elevate miR‐199a‐5p expression in ectopic endometrial mesenchymal stem cells. Up‐regulation of miR‐199a‐5p suppressed cell proliferation, motility and angiogenesis of these ectopic stem cells by targeting the 3′ untranslated region of VEGFA. Furthermore, we established an animal model of endometriosis and found that miR‐199a‐5p could decrease the size of endometriotic lesions in vivo. Taken together, this newly identified miR‐199a‐5p module provides a new avenue to the understanding of the processes of endometriosis development, especially proliferation, motility and angiogenesis, and may facilitate the development of potential therapeutics against endometriosis. Copyright

Leptin Affects Pregnancy Outcome of In Vitro Fertilization and Steroidogenesis of Human Granulosa Cells

Purpose: This study was to examine the serum leptin levels in the prediction of pregnancy outcomes in women receiving ovarian hyperstimulation. Effect of leptin on the steroidogenesis was evaluated for granulosa cell (GC) culture in vitro. Method: Serum levels of leptin and estradiol were measured on Day 2, the time of hCG administration and oocyte retrieval in 50 women undergoing long-course GnRH agonist ovarian hyperstimulation. The production of estrogen and progesterone in granulosa cell culture were detected after administration of leptin at the absence or presence of FSH 1 mIU. Results: Leptin levels at the time of hCG injection were significantly lower in the pregnant women than in those without pregnancy. Estradiol concentrations were not correlated with leptin levels during the time of hCG injection and oocyte retrieval. High leptin concentration suppressed both basal estradiol and progesterone productions in GC. Leptin in high doses suppressed estradiol production, but did not alter progesterone production of GC in the presence of FSH. Conclusions: Serum leptin levels may be predictive of IVF pregnancy outcome with the effect on intraovarian progesterone/estradiol ratio during the follicular phase. Significantly low serum leptin levels were noted in the pregnant women than in the nonpregnant women.

Increasing CD44+/CD24- tumor stem cells, and upregulation of COX-2 and HDAC6, as major functions of HER2 in breast tumorigenesis

BackgroundCancer cells are believed to arise primarily from stem cells. CD44+/CD24- have been identified as markers for human breast cancer stem cells. Although, HER2 is a well known breast cancer oncogene, the mechanisms of action of this gene are not completely understood. Previously, we have derived immortal (M13SV1), weakly tumorigenic (M13SV1R2) and highly tumorigenic (M13SV1R2N1) cell lines from a breast epithelial cell type with stem cell phenotypes after successive SV40 large T-antigen transfection, X-ray irradiation and ectopic expression of HER2/C-erbB2/neu. Recently, we found that M13SV1R2 cells became non-tumorigenic after growing in a growth factor/hormone-deprived medium (R2d cells).ResultsIn this study, we developed M13SV1R2N1 under the same growth factor/hormone-deprived condition (R2N1d cells). This provides an opportunity to analyze HER2 effect on gene expression associated with tumorigenesis by comparative study of R2d and R2N1d cells with homogeneous genetic background except HER2 expression. The results reveal distinct characters of R2N1d cells that can be ascribed to HER2: 1) development of fast-growing tumors; 2) high frequency of CD44+/CD24- cells (~50% for R2N1d vs. ~10% for R2d); 3) enhanced expression of COX-2, HDAC6 mediated, respectively, by MAPK and PI3K/Akt pathways, and many genes associated with inflammation, metastasis, and angiogenesis. Furthermore, HER2 expression can be down regulated in non-adhering R2N1d cells. These cells showed longer latent period and lower rate of tumor development compared with adhering cells.ConclusionsHER2 may induce breast cancer by increasing the frequency of tumor stem cells and upregulating the expression of COX-2 and HDAC6 that play pivotal roles in tumor progression.

Serum Adiponectin Levels Increase after Human Chorionic Gonadotropin Treatment during in vitro Fertilization

Aims: To determine whether or not serum adiponectin concentrations are influenced by ovarian hyperstimulation during in vitro fertilization (IVF). Methods: This study involved 52 women who were participating in IVF-ET cycles. Adiponectin levels in serum were determined by radioimmunoassay and compared. Results: Serum adiponectin levels fell from Day-basal to Day-hCG (p = 0.047), and then rose on Day-OR and again on Day-7ET (p < 0.001; p < 0.001). Estradiol levels on Day-hCG were significantly and positively correlated with serum adiponectin levels on Day-OR and Day-7ET (r = 0.325, p = 0.019; r = 0.372, p = 0.007). Progesterone levels on Day-OR positively correlated with serum adiponectin levels on Day-basal (r = 0.278, p = 0.046). There was also a positive correlation between progesterone levels on Day-7ET and serum adiponectin levels on Day-OR (r = 0.289, p = 0.038). Multiple linear regression analysis revealed that adiponectin levels on Day-OR and Day-7ET were negatively correlated with age and body mass index after adjustment was made for concomitant diseases. Conclusions: To sum up, following gonadotropin treatment, serum adiponectin levels decrease as a result of the negative effect of high estradiol levels on adiponectin production. Conversely, serum adiponectin levels increase following human chorionic gonadotropin treatment.

Laparoscopically Assisted Vaginal Hysterectomy versus Total Abdominal Hysterectomy: A Study of 100 Cases on Light-Endorsed Transvaginal Section

The objective of this study was to compare the results of a modified laparoscopically assisted vaginal hysterectomy (LAVH) procedure, using light-endorsed transvaginal section by two puncture trocars, with those of total abdominal hysterectomy (TAH) in a prospective, randomized, short-term study. A new, modified LAVH technique using Endo GIA stapler and two puncture trocars was established. For the laparoscopic phase, each adnexum was dissected, and the vesicouterine junction was identified clearly with the laparoscopic light from the vaginal side. Vaginal-phase surgery was performed as usual. Two hundred patients scheduled for abdominal hysterectomy were randomized to either LAVH (n = 100) or TAH (n = 100). Duration of hospitalization, time of surgery, dose of analgesics, and rates of complications were significantly lower in the LAVH group (p < 0.001). The average operating time was 77 ± 30 min for LAVH and 102 ± 18 min for TAH. The duration of hospitalization was 3.2 ± 0.7 days for LAVH and 5.5 ± 1.3 days for TAH. There were three complications in the LAVH group and 15 in the TAH group. Postoperative meperidine requirements (1.2 vs. 3.7 ampoules, 1 ampoule = 50 mg) were significantly fewer in the LAVH group. Regarding the training time, the mean operating time in the first 20 cases was 98 min, and in the last 20 cases it was 70.9 min. As compared with TAH and other modified LAVH procedures reported previously, the present technique is easy to learn and timesaving with fewer complications.

Targeted next-generation sequencing for molecular diagnosis of endometriosis-associated ovarian cancer

Recent molecular and pathological studies suggest that endometriosis may serve as a precursor of ovarian cancer (endometriosis-associated ovarian cancer, EAOC), especially of the endometrioid and clear cell subtypes. Accordingly, this study had two cardinal aims: first, to obtain mutation profiles of EAOC from Taiwanese patients; and second, to determine whether somatic mutations present in EAOC can be detected in preneoplastic lesions. Formalin-fixed paraffin-embedded (FFPE) tissues were obtained from ten endometriosis patients with malignant transformation. Macrodissection was performed to separate four different types of cells from FFPE sections in six patients. The four types of samples included normal endometrium, ectopic endometriotic lesion, atypical endometriosis, and carcinoma. Ultra-deep (>1000×) targeted sequencing was performed on 409 cancer-related genes to identify pathogenic mutations associated with EAOC. The most frequently mutated genes were PIK3CA (6/10) and ARID1A (5/10). Other recurrently mutated genes included ETS1, MLH1, PRKDC (3/10 each), and AMER1, ARID2, BCL11A, CREBBP, ERBB2, EXT1, FANCD2, MSH6, NF1, NOTCH1, NUMA1, PDE4DIP, PPP2R1A, RNF213, and SYNE1 (2/10 each). Importantly, in five of the six patients, identical somatic mutations were detected in atypical endometriosis and tumor lesions. In two patients, genetic alterations were also detected in ectopic endometriotic lesions, indicating the presence of genetic alterations in preneoplastic lesion. Genetic analysis in preneoplastic lesions may help to identify high-risk patients at early stage of malignant transformation and also shed new light on fundamental aspects of the molecular pathogenesis of EAOC.Key messagesMolecular characterization of endometriosis-associated ovarian cancer genes by targeted NGS.Candidate genes predictive of malignant transformation were identified.Chromatin remodeling, PI3K-AKT-mTOR, Notch signaling, and Wnt/β-catenin pathway may promote cell malignant transformation.

Elevated amniotic fluid leptin levels in early second trimester are associated with earlier delivery and lower birthweight in twin pregnancy

Objective.  To explore the possibility of using early second trimester amniotic fluid leptin levels as a predictor of pregnancy outcome in twin pregnancy.