Chii Ruey Tzeng

Assessment of uterine receptivity by the endometrial-subendometrial blood flow distribution pattern in women undergoing in vitro fertilization-embryo transfer ☆

OBJECTIVE To investigate the correlation of blood flow detected by color Doppler sonography in the endometrial-subendometrial unit with pregnancy outcome of IVF-ET treatments. DESIGN Prospective clinical study. SETTING University setting. PATIENT(S) Six hundred twenty-three patients selected prospectively on the day of ET. INTERVENTION(S) Transvaginal ultrasound examination was performed before ET. MAIN OUTCOME MEASURE(S) Association between pregnancy outcome and distribution of endometrial-subendometrial blood flow (primary outcome measure) and between pregnancy rate and endometrial measurements as well as uterine arterial blood flow (secondary outcome measures). RESULT(S) The overall pregnancy rate was 28.4% (177/623) per ET. The pregnancy and implantation rates of patients with the presence of both endometrial and subendometrial flow were 47.8% (64/134) and 24.2% (94/388); for patients with subendometrial flow only, 29.7% (102/343) and 15.8% (153/967); and for patients with no detectable endometrial-subendometrial flow, 7.5% (11/146) and 3.5% (13/376), respectively. The presence of both endometrial and subendometrial blood flow is indicative of good endometrial receptivity, whereas the absence of both represents a poor uterine environment. Nondetectable endometrial-subendometrial flow was associated with women who were older, had a thinner endometrium, and had higher uterine arterial resistance compared with those women who had detectable flow. CONCLUSION(S) Endometrial-subendometrial blood flow distribution pattern assessed by transvaginal color Doppler before ET is correlated with the implantation and pregnancy rate of IVF treatment.

Nitric oxide as a regulator in preimplantation embryo development and apoptosis

OBJECTIVE To investigate the mechanisms of nitric oxide (NO) in the development and apoptosis of preimplantation mouse embryos. DESIGN Prospective, controlled study. SETTING Medical college laboratory. SUBJECT(S) Two-cell embryos from outbred ICR mice. INTERVENTION(S) Hyperstimulation protocol, two-cell embryos were collected, then treated with or without an NO synthase inhibitor (L-NAME) or an NO donor (SNP) and combined with a cGMP analogue (8-Br-cGMP) or a selective inhibitor of NO-sensitive soluble guanylyl cyclase (ODQ). MAIN OUTCOME MEASURE(S) The development of ICR mouse embryo from two cells to blastocyst stages in vitro. RESULT(S) The development of blastocyst was inhibited by L-NAME in a concentration-dependent manner (0.1-10 microM) and 0.1 microM SNP reversed this effect (80.5% of control). Annexin-V/propidium iodide and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling techniques demonstrated that excessive NO (> or =10 microM) might induce apoptosis in the mouse embryos. 8-Br-cGMP reversed the inhibitory effect of L-NAME and rescued the embryo growth. ODQ inhibited the embryo development in a dose-responsive fashion (0.1--100 microM) but had no effect in the NO-induced embryo apoptosis. P53 and Bax were found to be up-regulated during the embryo fragmentation. CONCLUSION(S) These results indicate that the cGMP pathway might be involved in the NO-regulated embryonic development, but not in NO-induced apoptosis, for which P53/Bax pathway might be involved.

Antimüllerian hormone and polycystic ovary syndrome

OBJECTIVE To assess the relationship between antimüllerian hormone (AMH) and parameters related to polycystic ovary syndrome (PCOS). DESIGN Prospective study. SETTING Academic tertiary care center. PATIENT(S) A total of 290 women. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Parameters related to insulin resistance and metabolic syndrome. RESULT(S) Women with polycystic ovary morphology had significantly higher AMH levels than women in the control group. The prevalence of PCOS increased from 21% in the low-AMH (<4 ng/mL) group to 37% in the moderate-AMH (4-11 ng/mL) group and 80% in the high-AMH (>11 ng/mL) group. However, significant differences in insulin resistance parameters were not observed among groups. The results of the correlation analysis revealed that AMH levels were positively correlated with LH, total T, A, and total cholesterol content; however, AMH levels were negatively correlated with age, body mass index, and the number of menstrual cycles per year. AMH levels were not correlated with insulin resistance parameters. CONCLUSION(S) Elevated serum AMH levels increase the risk of PCOS but do not affect the risk of insulin resistance or metabolic syndrome.

Assessment of changes in utero‐ovarian arterial impedance during the peri‐implantation period by Doppler sonography in women undergoing assisted reproduction

To investigate changes in utero‐ovarian blood flow during the peri‐implantation period and their significance in successful embryo implantation.

Abnormal mitochondrial structure in human unfertilized oocytes and arrested embryos

Abstract: To clarify the relationship between mitochondria and embryo development, we collected human unfertilized oocytes, early embryos, and arrested embryos. Unfertilized oocytes and poor‐quality embryos were collected, and the ultrastructure of mitochondria was determined by transmission electron micrography. Four criteria for determining the mitochondrial state were mitochondrial morphology, cristae shape, location, and number of mitochondria. In mature oocytes, mitochondria were rounded with arched cristae and a dense matrix and were distributed evenly in the ooplasm. In pronuclear zygotes, the size and shape of mitochondria were similar to those in mature oocytes; however, mitochondria appeared to migrate and concentrate around pronuclei. In this study, 67% of examined unfertilized oocytes had fewer mitochondria in the cytoplasm. A decreased number of mitochondria located near the nucleus was also demonstrated in 60% of arrested embryos. Fewer differentiated cristae were determined in all three arrested blastocyst stages of embryos. The relative expressions of oxidative phosphorylation genes in oocytes and embryos were also determined. These data imply that inadequate redistribution of mitochondria, unsuccessful mitochondrial differentiation, or decreased mitochondrial transcription may result in poor oocyte fertilization and compromised embryo development.

Multiple rearrangements of mitochondrial DNA in unfertilized human oocytes

OBJECTIVE To determine the rearrangement of mitochondrial DNA (mtDNA) in unfertilized human oocytes and compromised embryos to evaluate the fertilization capacity of oocytes. DESIGN Prospective laboratory research. SETTING IVF laboratory in a university hospital. PATIENT(S) One hundred twenty-four unfertilized oocytes, 98 arrested embryos, and 45 tripronucleate (3PN) embryos from 65 female patients undergoing in vitro fertilization (IVF). INTERVENTION(S) Unfertilized oocytes and poor quality embryos were collected 48 hours after IVF. MAIN OUTCOME MEASURE(S) Comparison of the frequency of mtDNA deletions and fertilization rates of oocytes. RESULT(S) Multiple deletions of mtDNA were found in unfertilized oocytes and arrested embryos obtained from IVF patients. A 4977-bp deletion was the most frequent deletion in human oocytes and embryos. About 66.1% of the unfertilized oocytes, 34.8% of the arrested or fragmented embryos, and 21.1% of the 3PN embryos harbored the 4977-bp deletion of mtDNA. There was a significant increase in the proportion of deleted mtDNA in unfertilized oocytes. CONCLUSION(S) Accumulation of mtDNA deletions may contribute to mitochondrial dysfunction and impaired ATP production. We conclude that the accumulation of rearranged mtDNA may interfere with fertilization of human oocytes and further embryonic development.

miRNA-34b as a tumor suppressor in estrogen-dependent growth of breast cancer cells

IntroductionEstrogen is involved in several physiological and pathological processes through estrogen receptor (ER)-mediated transcriptional gene regulation. miRNAs (miRs), which are noncoding RNA genes, may respond to estrogen and serve as posttranscriptional regulators in tumorigenic progression, especially in breast cancer; however, only limited information about this possibility is available. In the present study, we identified the estrogen-regulated miR-34b and investigated its functional role in breast cancer progression.MethodsEstrogen-regulated miRNAs were identified by using a TaqMan low density array. Our in vivo Tet-On system orthotopic model revealed the tumor-suppressive ability of miR-34b. Luciferase reporter assays and chromatin immunoprecipitation assay demonstrated miR-34b were regulated by p53-ER interaction.ResultsIn this study, we identified one such estrogen downregulated miRNA, miR-34b, as an oncosuppressor that targets cyclin D1 and Jagged-1 (JAG1) in an ER+/wild-type p53 breast cancer cell line (MCF-7), as well as in ovarian and endometrial cells, but not in ER-negative or mutant p53 breast cancer cell lines (T47D, MBA-MB-361 and MDA-MB-435). There is a negative association between ERα and miR-34b expression levels in ER+ breast cancer patients. Tet-On induction of miR-34b can cause inhibition of tumor growth and cell proliferation. Also, the overexpression of miR-34b inhibited ER+ breast tumor growth in an orthotopic mammary fat pad xenograft mouse model. Further validation indicated that estrogens inhibition of miR-34b expression was mediated by interactions between ERα and p53, not by DNA methylation regulation. The xenoestrogens diethylstilbestrol and zeranol also showed similar estrogenic effects by inhibiting miR-34b expression and by restoring the protein levels of the miR-34b targets cyclin D1 and JAG1 in MCF-7 cells.ConclusionsThese findings reveal that miR-34b is an oncosuppressor miRNA requiring both ER+ and wild-type p53 phenotypes in breast cancer cells. These results improve our ability to develop new therapeutic strategies to target the complex estrogenic pathway in human breast cancer progression through miRNA regulation.

Clinical and biochemical presentation of polycystic ovary syndrome in women between the ages of 20 and 40

BACKGROUND The clinical features and metabolic complications of polycystic ovary syndrome (PCOS) may change with age. This study was designed to investigate the clinical and biochemical characteristics of PCOS patients between the ages of 20 and 40. METHODS The study included 781 Taiwanese women, of whom 453 were diagnosed with PCOS and 328 were non-PCOS controls. Anthropometric components, androgens, endocrine, insulin resistance, and metabolic components were measured and correlated with age. Above parameters were compared between younger and elder women with PCOS. RESULTS Age had significant negative correlations with androgens (total testosterone and dehydroepiandrosterone sulfate), the modified Ferriman-Gallwey score and the prevalence of acne and hirsutism. Age had significant positive correlations with fasting glucose, cholesterol, triglycerides and low-density lipoprotein. The 453 women who fulfilled diagnostic criteria for PCOS were classified by age into two groups: Group A (20-29 years old, n= 294) and Group B (30-40 years old, n= 159). Group A had significantly higher total testosterone levels than Group B. Group B had higher fasting insulin and glucose levels, triglycerides, body mass index and waist measurements and a higher incidence of obesity than Group A. The average ovarian volume was not significantly different among the two groups. CONCLUSIONS Increased age is accompanied by a decrease in the prevalence of both clinical and biochemical hyperandrogenism in women. Hyperandrogenism is the important factor for young women with PCOS; however, abdominal obesity and certain metabolic disturbances became major concerns for older women with PCOS.

Expression of the pluripotent transcription factor OCT4 promotes cell migration in endometriosis.

OBJECTIVE To identify the impact of the pluripotent transcription factor OCT4 in endometrial cell migration and endometriosis. DESIGN The OCT4 expression and cell migration study. SETTING Research institution and reproductive medical clinic. PATIENT(S) Nine subjects with normal endometrium, 3 subjects with normal myometrium, 36 patients with hyperplastic endometrium, and 58 patients with endometriosis. INTERVENTION(S) The expression of OCT4 messenger RNA in normal endometrium, normal myometrium, hyperplastic endometrium, and ectopic endometriotic tissues was analyzed using reverse transcription and quantitative real-time polymerase chain reaction (PCR). The effect of OCT4 expression on the migration activity of the endometrial cells was examined. MAIN OUTCOME MEASURE(S) Reverse transcription and quantitative real-time PCR, Western blotting, and wound closure and transwell assays. RESULT(S) The expression of OCT4 and NANOG messenger RNA was significantly higher in ectopic endometriotic tissues, compared with that of the normal endometrium, the normal myometrium, and the hyperplastic endometrium. The level of OCT4 messenger RNA in endometriotic tissues was positively correlated with the expression of genes associated with cell migration. Overexpression of the OCT4 protein in primary human endometriotic stromal cells and human RL95-2 and HEC1A endometrial carcinoma cell lines resulted in decreased levels of E-CADHERIN, the increased expression of the VIMENTIN, TWIST, and SLUG proteins, and an increase in the migration activity of endometrial cells in transwell and wound closure assays. CONCLUSION(S) The transcription of the OCT4 gene is significantly up-regulated in human ectopic endometriotic tissues. The expression of OCT4 may contribute to the pathology of ectopic endometrial growth by stimulating the migration activity of endometrial cells.

Effects of growth factors and granulosa cell co-culture on in-vitro maturation of oocytes

The maturation medium for in-vitro oocyte maturation is usually supplemented with serum. However, supplementation with serum from pregnant women adversely affects the outcome of in-vitro maturation. The purpose of the study was to assess if growth factors or granulosa cell co-culture could overcome the adverse effects of pregnant womens serum. The basal maturation medium consisted of TCM199, 75 mIU/ml human menopausal gonadotrophin (HMG), 0.2 mmol/l pyruvate, and 10% serum. The maturation medium for control 1 contained fertile Womens serum. The maturation medium for control 2 contained pregnant Womens serum. The maturation media for the study groups consisted of medium for control 2, with the addition of EGF, IGF-I, activin, TGFbeta or granulosa cell co-culture. Immature oocytes were obtained from FVB mice, and the experiment was repeated six times. After maturation, the oocytes were fertilized and cultured to blastocysts, and the cumulus cells were analysed for apoptosis. The maturation, fertilization and blastocyst rates of the control 2 group were significantly lower than those of control 1 group (P < 0.05). Addition of EGF, IGF-1, activin, TGFbeta or granulosa cell co-culture could not improve the outcome of in-vitro maturation. Cumulus cell proliferation was inhibited by pregnant womans serum. Apoptosis of cumulus cell was not related to in-vitro oocyte maturation and subsequent embryo development.