yuh-ming hwu - Academia.edu (original) (raw)

Papers by yuh-ming hwu

Research paper thumbnail of Dual trigger with gonadotropin releasing hormone agonist (GnRHa) and human chorionic gonadotropin (hCG) for the treatment of ‘immature oocyte syndrome’ (IOS)

Fertility and Sterility, 2012

funding and multiple pregnancy rates were significantly decreased. Further studies are needed to ... more funding and multiple pregnancy rates were significantly decreased. Further studies are needed to choose a viable embryo in order to transfer to reduce multiple-pregnancy rate even in the IVM program.

Research paper thumbnail of Intrauterine adhesion prevention with Malecot catheter after hysteroscopic myomectomy: A novel approach

Taiwanese Journal of Obstetrics and Gynecology, 2020

Objective: Intrauterine adhesion after hysteroscopic myomectomy contributes to infertility, recur... more Objective: Intrauterine adhesion after hysteroscopic myomectomy contributes to infertility, recurrent miscarriages, menstrual irregularities, and hinders pregnancy outcomes. The aim of this study was to apply the indwelling Malecot catheter in prevention of intrauterine adhesion after hysteroscopic myomectomy and to further evaluate the effectiveness of this approach with reported live birth rates in infertile patients who underwent subsequent infertility treatment. Materials and methods: Seventeen patients with FIGO Classification System PALM-COIEN Type 0 or 1 submucous myoma that received hysteroscopic myomectomy were recruited in this retrospective analysis. Post-operative insertion of the Malecot catheter via the aid of the uterine sound was performed and the catheter was left in place for seven days. Results: The mean duration of TTP (time to pregnancy) was 15.6 months after hysteroscopy. Within three years after the operation, 10 out of 17 infertility patients achieved ongoing pregnancy over 12 weeks. Ongoing pregnancy rate was 58.8% (10/17). Eight patients achieved live birth (seven singletons, one twin pregnancy) with mean gestational age of 38 weeks. Live birth rate was 47.1% (8/17). Conclusion: The Malecot catheter is an inexpensive, easy-to-operate, and effective physical barrier method for preventing IUA in infertile patients undergoing hysteroscopic myomectomy with high live birth rate and no obvious visible post-operative adhesions.

Research paper thumbnail of Survival Motor Neuron Protein Participates in Mouse Germ Cell Development and Spermatogonium Maintenance

International Journal of Molecular Sciences, 2020

The defective human survival motor neuron 1 (SMN1) gene leads to spinal muscular atrophy (SMA), t... more The defective human survival motor neuron 1 (SMN1) gene leads to spinal muscular atrophy (SMA), the most common genetic cause of infant mortality. We previously reported that loss of SMN results in rapid differentiation of Drosophila germline stem cells and mouse embryonic stem cells (ESCs), indicating that SMN also plays important roles in germ cell development and stem cell biology. Here, we show that in healthy mice, SMN is highly expressed in the gonadal tissues, prepubertal spermatogonia, and adult spermatocytes, whereas low SMN expression is found in differentiated spermatid and sperm. In SMA-like mice, the growth of testis tissues is retarded, accompanied with gamete development abnormalities and loss of the spermatogonia-specific marker. Consistently, knockdown of Smn1 in spermatogonial stem cells (SSCs) leads to a compromised regeneration capacity in vitro and in vivo in transplantation experiments. In SMA-like mice, apoptosis and accumulation of the R-loop structure were s...

Research paper thumbnail of Spatiotemporal Expression of the Serine Protease Inhibitor SERPINE2 in the Mouse Uterus During the Estrous Cycle, Pregnancy, and Lactation Period

Biology of Reproduction, 2010

Research paper thumbnail of Derivation of Patient Specific Pluripotent Stem Cells Using Clinically Discarded Cumulus Cells

PloS one, 2016

Induced pluripotent stem cells (iPSCs) are powerful tools for basic and translational research, a... more Induced pluripotent stem cells (iPSCs) are powerful tools for basic and translational research, as well as regenerative medicine. In routine human in vitro fertilization (IVF) practices, cumulus cells (CCs) are discarded, representing a potential source of biological materials for regenerative medicine. In this study, we derived patient-specific iPSCs using CCs from human infertility clinics for the first time. The human cumulus cell derived iPSCs (hc-iPSCs) were characterized for growth, karyotype, expression of pluripotency genes, and were subjected to embryoid bodies (EBs) and teratoma assays to evaluate their differentiation capacity. Hc-iPSCs display typical iPSC characteristics, and are capable of differentiating into all germ layers in vitro and in vivo. We further show that putative primordial germ cell like cells (PGCLCs) can be derived using hc-iPSCs. Our data demonstrate the feasibility of deriving patient-specific pluripotent stem cells using CCs.

Research paper thumbnail of VEGF and FGF2 Improve Revascularization, Survival, and Oocyte Quality of Cryopreserved, Subcutaneously-Transplanted Mouse Ovarian Tissues

International journal of molecular sciences, Jan 30, 2016

This study was conducted to investigate the effect of the vascular endothelial growth factor (VEG... more This study was conducted to investigate the effect of the vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) on revascularization, survival, and oocyte quality of cryopreserved, subcutaneously-transplanted mouse ovarian tissue. Autologous subcutaneous transplantation of vitrified-thawed mouse ovarian tissues treated with (experimental group) or without (control group) VEGF and FGF2 was performed. After transplantation to the inguinal region for two or three weeks, graft survival, angiogenesis, follicle development, and oocyte quality were examined after gonadotropin administration. VEGF coupled with FGF2 (VEGF/FGF2) promoted revascularization and significantly increased the survival rate of subcutaneously-transplanted cryopreserved ovarian tissues compared with untreated controls. The two growth factors did not show long-term effects on the ovarian grafts. In contrast to the untreated ovarian grafts, active folliculogenesis was revealed as the number of ...

Research paper thumbnail of Quantitative analysis of total β-subunit of human chorionic gonadotropin concentration in urine by immunomagnetic reduction to assist in the diagnosis of ectopic pregnancy

International journal of nanomedicine, 2015

The initial diagnosis of ectopic pregnancy depends on physical examination, ultrasound, and seria... more The initial diagnosis of ectopic pregnancy depends on physical examination, ultrasound, and serial measurements of total β-subunit of human chorionic gonadotropin (hCGβ) concentrations in serum. The aim of this study was to explore the possibility of using quantitative analysis of total hCGβ in urine rather than in serum by immunomagnetic reduction (IMR) assay as an alternative method to diagnose an ectopic pregnancy. We established a standard calibration curve of IMR intensity against total hCGβ concentration based on standard hCGβ samples, and used an IMR assay to detect total hCGβ concentrations in the urine of pregnant women with lower abdominal pain and/or vaginal bleeding. The final diagnosis of ectopic pregnancy was based on ultrasound scans, operative findings, and pathology reports. In this prospective study, ten clinical samples were used to analyze the relationship of total hCGβ IMR signals between urine and serum. Furthermore, 20 clinical samples were used to analyze the...

Research paper thumbnail of Concordant myelomeningocele in dizygotic twins conceived by intracytoplasmic sperm injection, in vitro fertilization, and embryo transfer

Taiwanese journal of obstetrics & gynecology, 2013

Research paper thumbnail of Embryonic early-cleavage rate is decreased with aging in GnRH agonist but not inantagonist protocols

Journal of assisted reproduction and genetics, Jan 20, 2015

The aim of this study was to evaluate the correlation between embryonic early-cleavage status and... more The aim of this study was to evaluate the correlation between embryonic early-cleavage status and the age of patients receiving either a GnRH agonist long protocol or a GnRH antagonist protocol. This retrospective study included 534 patients undergoing a fresh cycle of oocyte retrieval and day-3 embryo transfer. Of the 534 patients treated, 331 received a GnRH agonist long stimulation protocol (GnRH agonist group) for ovarian stimulation and 203 patients received a GnRH antagonist protocol (GnRH antagonist group). By logistic regression analysis, the rate of embryonic early-cleavage was significantly decreased with increasing age of women in the agonist…

Research paper thumbnail of The safety and efficacy of microwave endometrial ablation after endometrial curettage without hormonal pretreatment

Taiwanese journal of obstetrics & gynecology, 2007

To evaluate the safety and efficacy of a microwave endometrial ablation (MEA) procedure to treat ... more To evaluate the safety and efficacy of a microwave endometrial ablation (MEA) procedure to treat patients with menorrhagia after endometrial curettage but without medical pretreatment. From February to September 2001, 19 eligible and consenting premenopausal women with menorrhagia underwent endometrial curettage, immediately followed by MEA. No medical pretreatment with gonadotropin-releasing hormone (GnRH) analogues or danazol was given for endometrial preparation. The severity of menorrhagia was assessed using the menstrual score. The patients were followed up for at least 3 years. Of the 19 women, 17 had completed at least 3 years' follow-up. Fourteen patients (82%) were satisfied with the posttreatment menstrual status, comparable with those patients receiving medical pretreatment of other studies (78-94%). The mean menstrual score was 11.8 before treatment and 1.95, 2.25, 2.2, 2.1 and 2.1 at 3, 6, 12, 24 and 36 months posttreatment, respectively. Of the 12 patients who comp...

Research paper thumbnail of The efficiency of conventional microscopic selection is comparable to the hyaluronic acid binding method in selecting spermatozoa for male infertility patients

Taiwanese journal of obstetrics & gynecology, 2015

To evaluate if hyaluronic acid (HA)-bound spermatozoa surpassed conventional microscopy-selected ... more To evaluate if hyaluronic acid (HA)-bound spermatozoa surpassed conventional microscopy-selected spermatozoa in the status of sperm DNA integrity by acridine orange (AO) fluorescence staining. Spermatozoa obtained from couples with indication for the intracytoplasmic sperm injection (ICSI) procedure due to male infertility (n = 34) and control males with normal sperm parameters (n = 12) were analyzed using AO fluorescence staining after density-gradient centrifugation (DGC), polyvinylpyrrolidone (PVP)-microscopic selection, and HA-binding selection to determine sperm DNA integrity. Percentages of DNA intact spermatozoa with green fluorescence were significantly higher in both PVP-microscopic selected spermatozoa (82.1 ± 24.0%) and HA-bound spermatozoa (83.9 ± 21.1%) than in spermatozoa prepared by DGC (66.8 ± 24.0%). However, there was no significant difference between the PVP-sperm and HA-sperm groups. When the percentage of green fluorescent spermatozoa prepared by DGC fell initia...

Research paper thumbnail of Embryonic Dormancy Phenomenon in Obstructed Healthy Mouse Fallopian Tubes

A mouse model of unilateral oviductal obstruction was designed to study whether healthy oviducts ... more A mouse model of unilateral oviductal obstruction was designed to study whether healthy oviducts can support embryo development in an advanced stage toward blastocyst hatching and implantation when the embryos in the contralateral side normally move into the uterine cavity. Methods: The oviducts of 80 female ICR mice (aged 5-8 weeks) were ligated unilaterally 12-40 hr postcoitus. The ligated oviducts were isolated from day 4 to 19.5 postcoitus. Embryos within the ligated oviducts were then flushed out to record the developmental stage and compared with the conceptuses in the contralateral uterine horns with unligated oviducts. Embryos recovered from ligated oviducts were then cultured in vitro to observe their potential for further development. Results: In 33 mice, 53.4% (163/305) and 86.1% (241/280) of the morphologically normal blastocysts had hatched from the zona pellucida within the obstructed tube and contralateral uterine horns, respectively, on the 5th day postcoitus. The data demonstrated that the hatching process could take place within the obstructed fallopian tube, but the timing was delayed. From 5.5 to 19.5 days postcoitus, a total of 362 implanted embryos were obtained in unligated control

Research paper thumbnail of Mosaic Trisomy 7 at Amniocentesis: Prenatal Diagnosis and Molecular Genetic Analyses

Taiwanese Journal of Obstetrics and Gynecology, 2010

Objective: To present prenatal diagnosis and molecular genetic analyses of mosaic trisomy 7. Mate... more Objective: To present prenatal diagnosis and molecular genetic analyses of mosaic trisomy 7. Materials, Methods and Results: A 38-year-old primigravid woman underwent amniocentesis at 19 weeks of gestation because of her advanced maternal age. Amniocentesis revealed a karyotype of 47,XY,+7[26]/46, XY[16]. Repeated amniocentesis at 21 weeks of gestation revealed a karyotype of 47,XY,+7[20]/46,XY[17]. Simultaneous cordocentesis revealed a karyotype of 46,XY in 100/100 cultured lymphocytes. Polymorphic DNA marker analyses of uncultured amniocytes and cord blood revealed a diallelic pattern with seemingly equal biparental inheritance of chromosome 7. Repeated cordocentesis and chorionic villus sampling at 23 weeks of gestation revealed a karyotype of 47,XY,+7[2]/46,XY[66] in cord blood and a karyotype of 47,XY,+7 in 24/24 cultured chorionic villi cells. Level II ultrasonography was normal. At 40 weeks of gestation, a 2,708 g normal male baby was delivered. The peripheral blood had a karyotype of 46,XY in 100/100 lymphocytes. Molecular analyses of placenta, urine, buccal swab, and peripheral blood revealed a diallelic pattern and seemingly equal biparental inheritance of chromosome 7 in all tissues. At 3 months of age, he manifested hypopigmented skin and inguinal hernia, but showed normal growth and mental development. Fluorescence in situ hybridization analysis of inguinal hernia sac tissue revealed that 19/100 (19%) of nuclei had three chromosome 7 signals. Conclusion: Mosaic trisomy 7 at amniocentesis may be derived from a cell culture artifact from an undetected low level of trisomy 7 mosaicism in uncultured amniocytes, and can be associated with favorable fetal outcome if the blood has a normal karyotype or a very low level of mosaicism and if uniparental disomy for chromosome 7 is excluded.

Research paper thumbnail of Clinical application of immunomagnetic reduction for quantitative measurement of insulin-like growth factor binding protein-1 in the prediction of pregnant women with preterm premature rupture of membranes

Clinica Chimica Acta, 2015

Background: The most important factor for a successful pregnancy after in vitro fertilization is ... more Background: The most important factor for a successful pregnancy after in vitro fertilization is embryo quality. The aim of this study was to explore the possibility that using the immunomagnetic reduction (IMR) assay to quantitatively measure β-subunit of human chorionic gonadotropin (β-hCG) in blastocyst culture media to differentiate embryo quality. Methods: This was a prospective case-control study including 28 samples of blastocyst culture media. We used single-step blastocyst culture and IMR assay to analyze β-hCG concentrations in culture media. We also explored the relationship between IMR signals of β-hCG and morphological grading of blastocysts. Results: β-hCG concentration-dependent IMR signals were highly correlated with blastocyst morphological quality (Spearman correlation coefficient: 0.731). Receiver-operating characteristic curve analysis showed a cutoff IMR value to differentiate embryo quality of 0.873%, with an area under the curve of 0.947, sensitivity of 0.882 and specificity of 0.818. Furthermore, subanalysis also revealed a positive correlation between β-hCG concentration-dependent IMR signals and trophectoderm grading, with a Spearman correlation coefficient of 0.576. Conclusions: An IMR assay can quantitatively measure β-hCG in blastocyst culture media, and may be a potential clinical tool to assist in the assessment of good blastocyst quality before embryo transfer.

Research paper thumbnail of Differential expression and distribution of alternatively spliced transcripts of PDGF-A and of PDGF receptor-alpha in mouse reproductive tissues

Life sciences, Jan 23, 2005

Platelet-derived growth factor-A (PDGF-A) affects cellular activities such as proliferation, diff... more Platelet-derived growth factor-A (PDGF-A) affects cellular activities such as proliferation, differentiation, and development by way of paracrine or autocrine interaction with PDGF-A receptor alpha (PDGFR-alpha). Two forms of alternatively spliced PDGF-A mRNA, a long and a short isoform, have been found in several mammalian species. Expression of PDGF-A and its cognate receptor PDGFR-alpha has been well studied in various tissues. However, these investigations did not distinguish between the individual isoforms of PDGF-A. In the present investigation, we identified the differential cellular expression patterns of the two isoforms of PDGF-A and of PDGFR-alpha in mouse reproductive tissues by using laser capture microdissection coupled with reverse transcriptase polymerase chain reaction. The long PDGF-A mRNA isoform was primarily detected in the epithelium, while the short isoform was ubiquitously distributed in epithelium, stroma, and muscle cells, although it was still more promine...

Research paper thumbnail of Relationship of follicular size to the development of intracytoplasmic sperm injection-derived human embryos

Taiwanese journal of obstetrics & gynecology, 2010

To compare the embryonic development of oocytes obtained from follicles of different sizes. Oocyt... more To compare the embryonic development of oocytes obtained from follicles of different sizes. Oocytes (n = 819) were retrieved from women at 40 years of age or younger during 86 in vitro fertilization cycles and categorized as small, medium, or large based on the estimated volume of follicular fluid at the time of retrieval. The rates of good quality embryos from the large, medium, and small groups on days 2 and 3 were 76.85% and 66.20%, 74.00% and 61.33%, and 69.81% and 58.49%, respectively. There were no significant differences in the rates of good quality embryos between the three follicular volume groups. Even though fewer oocytes completed maturation in the small follicle group than in the other two groups, the quality of the embryos in all three groups was the same on days 2 and 3. These findings suggest that follicles of all sizes should be aspirated during the intracytoplasmic sperm injection cycle as follicles of every size were a good source of embryos.

Research paper thumbnail of Comparison of day 2 tubal embryo transfer with day 3 embryo transfer into the uterus of intracytoplasmic sperm injection derived human embryos

Taiwanese journal of obstetrics & gynecology, 2006

To compare the results of embryo transfer into the uterus (ET) performed on day 3 vs. tubal embry... more To compare the results of embryo transfer into the uterus (ET) performed on day 3 vs. tubal embryo transfer (TET) on day 2 in couples diagnosed with male factor infertility. We collected data from 34 and 110 patients who underwent intracytoplasmic sperm injection (ICSI) followed by either TET on day 2 or ET on day 3 (January 2001-June 2005), respectively. All couples were diagnosed with male factor infertility; there were no tubal or uterine factors, diminished ovarian reserve, pelvic adhesions, history of polycystic ovary syndrome or previous ovarian surgery. The clinical pregnancy rates of day 2 TET and day 3 ET following ICSI were 64.71% and 57.27% (p > 0.05), respectively. The corresponding implantation rates were 35.47% and 29.58% (p > 0.05), respectively. The implantation and clinical pregnancy rates of ET on the 3rd day following ICSI were similar to the rates obtained with TET 2 days after ICSI. Therefore, ET performed 3 days after ICSI should be the first choice for c...

Research paper thumbnail of Early-cleavage is a reliable predictor for embryo implantation in the GnRH agonist protocols but not in the GnRH antagonist protocols

Reproductive Biology and Endocrinology, 2009

Background To test if early-cleavage was a strong predictor of pregnancy in patients receiving ei... more Background To test if early-cleavage was a strong predictor of pregnancy in patients receiving either a GnRH agonist long protocol or a GnRH antagonist protocol for in-vitro fertilization treatment (IVF) and intracytoplasmic sperm injection (ICSI). Methods This retrospective study included 534 patients undergoing a fresh cycle of oocyte retrieval and the day-3 embryo transfer (from 22 to 46 years old). Of the 534 patients treated, 331 received a GnRH agonist long stimulation protocol (GnRH agonist group) for ovarian stimulation and 203 patients received a GnRH antagonist protocol (GnRH antagonist group). In each group, patients who had at least one early-cleavage embryo transferred were designated as the 'early-cleavage' subgroup. Patients who had no early-cleavage embryos transferred were designated as the 'late-cleavage' subgroup. Results The early cleavage rate was significantly lower in the GnRH antagonist group compared with that in the GnRH agonist group (IVF c...

Research paper thumbnail of Prenatal identification of fetal overgrowth, abdominal wall defect, and neural tube defect in pregnancies achieved by assisted reproductive technology

Research paper thumbnail of Involvement of the Serine Protease Inhibitor, SERPINE2, and the Urokinase Plasminogen Activator in Cumulus Expansion and Oocyte Maturation

PLoS ONE, 2013

The serpin peptidase inhibitor, clade E, member 2 (SERPINE2) inhibits urokinase-type plasminogen ... more The serpin peptidase inhibitor, clade E, member 2 (SERPINE2) inhibits urokinase-type plasminogen activator (PLAU) and tissue-type plasminogen activator. Higher SERPINE2 expression levels were detected in cumulus cells of human immature oocytes than in those of mature oocytes. The objective of this study was to evaluate whether high SERPINE2 levels in cumulus cells are associated with oocyte immaturity. Using the mouse cumulus-oocyte complex as an experimental model, the effects of elimination and overexpression of SERPINE2 in cumulus cells on cumulus expansion and oocyte maturation were assayed by in vitro maturation. Serpine2 and PLAU transcripts were the most highly expressed serpins and plasminogen activators, respectively. Their expression was coordinately regulated in cumulus cells during gonadotropin-induced oocyte maturation. Silencing of Serpine2 expression using small interfering RNAs or blockage of SERPINE2 protein using a specific antibody had no effect on oocyte maturation. However, overexpression of Serpine2 or exogenous supplementation with high levels of SERPINE2 impaired cumulus expansion and oocyte maturation, probably by decreasing hyaluronan synthase 2 (Has2) and versican (Vcan) mRNA expression. Amiloride, a specific PLAU inhibitor, also suppressed these processes. PLAU supplementation of the oocyte in vitro maturation medium caused earlier and more extensive expansion of cumulus cells and oocyte maturation that may be mediated by increased Has2 mRNA expression. However, these effects were neutralized by coincubation with SERPINE2 or amiloride and PLAU. In conclusion, SERPINE2 and PLAU are involved in cumulus expansion and oocyte maturation. High SERPINE2 levels impair these processes, probably by decreasing cumulus matrix gene expression as well as reducing cumulus hyaluronan contents and inhibiting PLAU activity. These findings may explain why cumulus cells surrounding immature human oocytes express high SERPINE2 levels.

Research paper thumbnail of Dual trigger with gonadotropin releasing hormone agonist (GnRHa) and human chorionic gonadotropin (hCG) for the treatment of ‘immature oocyte syndrome’ (IOS)

Fertility and Sterility, 2012

funding and multiple pregnancy rates were significantly decreased. Further studies are needed to ... more funding and multiple pregnancy rates were significantly decreased. Further studies are needed to choose a viable embryo in order to transfer to reduce multiple-pregnancy rate even in the IVM program.

Research paper thumbnail of Intrauterine adhesion prevention with Malecot catheter after hysteroscopic myomectomy: A novel approach

Taiwanese Journal of Obstetrics and Gynecology, 2020

Objective: Intrauterine adhesion after hysteroscopic myomectomy contributes to infertility, recur... more Objective: Intrauterine adhesion after hysteroscopic myomectomy contributes to infertility, recurrent miscarriages, menstrual irregularities, and hinders pregnancy outcomes. The aim of this study was to apply the indwelling Malecot catheter in prevention of intrauterine adhesion after hysteroscopic myomectomy and to further evaluate the effectiveness of this approach with reported live birth rates in infertile patients who underwent subsequent infertility treatment. Materials and methods: Seventeen patients with FIGO Classification System PALM-COIEN Type 0 or 1 submucous myoma that received hysteroscopic myomectomy were recruited in this retrospective analysis. Post-operative insertion of the Malecot catheter via the aid of the uterine sound was performed and the catheter was left in place for seven days. Results: The mean duration of TTP (time to pregnancy) was 15.6 months after hysteroscopy. Within three years after the operation, 10 out of 17 infertility patients achieved ongoing pregnancy over 12 weeks. Ongoing pregnancy rate was 58.8% (10/17). Eight patients achieved live birth (seven singletons, one twin pregnancy) with mean gestational age of 38 weeks. Live birth rate was 47.1% (8/17). Conclusion: The Malecot catheter is an inexpensive, easy-to-operate, and effective physical barrier method for preventing IUA in infertile patients undergoing hysteroscopic myomectomy with high live birth rate and no obvious visible post-operative adhesions.

Research paper thumbnail of Survival Motor Neuron Protein Participates in Mouse Germ Cell Development and Spermatogonium Maintenance

International Journal of Molecular Sciences, 2020

The defective human survival motor neuron 1 (SMN1) gene leads to spinal muscular atrophy (SMA), t... more The defective human survival motor neuron 1 (SMN1) gene leads to spinal muscular atrophy (SMA), the most common genetic cause of infant mortality. We previously reported that loss of SMN results in rapid differentiation of Drosophila germline stem cells and mouse embryonic stem cells (ESCs), indicating that SMN also plays important roles in germ cell development and stem cell biology. Here, we show that in healthy mice, SMN is highly expressed in the gonadal tissues, prepubertal spermatogonia, and adult spermatocytes, whereas low SMN expression is found in differentiated spermatid and sperm. In SMA-like mice, the growth of testis tissues is retarded, accompanied with gamete development abnormalities and loss of the spermatogonia-specific marker. Consistently, knockdown of Smn1 in spermatogonial stem cells (SSCs) leads to a compromised regeneration capacity in vitro and in vivo in transplantation experiments. In SMA-like mice, apoptosis and accumulation of the R-loop structure were s...

Research paper thumbnail of Spatiotemporal Expression of the Serine Protease Inhibitor SERPINE2 in the Mouse Uterus During the Estrous Cycle, Pregnancy, and Lactation Period

Biology of Reproduction, 2010

Research paper thumbnail of Derivation of Patient Specific Pluripotent Stem Cells Using Clinically Discarded Cumulus Cells

PloS one, 2016

Induced pluripotent stem cells (iPSCs) are powerful tools for basic and translational research, a... more Induced pluripotent stem cells (iPSCs) are powerful tools for basic and translational research, as well as regenerative medicine. In routine human in vitro fertilization (IVF) practices, cumulus cells (CCs) are discarded, representing a potential source of biological materials for regenerative medicine. In this study, we derived patient-specific iPSCs using CCs from human infertility clinics for the first time. The human cumulus cell derived iPSCs (hc-iPSCs) were characterized for growth, karyotype, expression of pluripotency genes, and were subjected to embryoid bodies (EBs) and teratoma assays to evaluate their differentiation capacity. Hc-iPSCs display typical iPSC characteristics, and are capable of differentiating into all germ layers in vitro and in vivo. We further show that putative primordial germ cell like cells (PGCLCs) can be derived using hc-iPSCs. Our data demonstrate the feasibility of deriving patient-specific pluripotent stem cells using CCs.

Research paper thumbnail of VEGF and FGF2 Improve Revascularization, Survival, and Oocyte Quality of Cryopreserved, Subcutaneously-Transplanted Mouse Ovarian Tissues

International journal of molecular sciences, Jan 30, 2016

This study was conducted to investigate the effect of the vascular endothelial growth factor (VEG... more This study was conducted to investigate the effect of the vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) on revascularization, survival, and oocyte quality of cryopreserved, subcutaneously-transplanted mouse ovarian tissue. Autologous subcutaneous transplantation of vitrified-thawed mouse ovarian tissues treated with (experimental group) or without (control group) VEGF and FGF2 was performed. After transplantation to the inguinal region for two or three weeks, graft survival, angiogenesis, follicle development, and oocyte quality were examined after gonadotropin administration. VEGF coupled with FGF2 (VEGF/FGF2) promoted revascularization and significantly increased the survival rate of subcutaneously-transplanted cryopreserved ovarian tissues compared with untreated controls. The two growth factors did not show long-term effects on the ovarian grafts. In contrast to the untreated ovarian grafts, active folliculogenesis was revealed as the number of ...

Research paper thumbnail of Quantitative analysis of total β-subunit of human chorionic gonadotropin concentration in urine by immunomagnetic reduction to assist in the diagnosis of ectopic pregnancy

International journal of nanomedicine, 2015

The initial diagnosis of ectopic pregnancy depends on physical examination, ultrasound, and seria... more The initial diagnosis of ectopic pregnancy depends on physical examination, ultrasound, and serial measurements of total β-subunit of human chorionic gonadotropin (hCGβ) concentrations in serum. The aim of this study was to explore the possibility of using quantitative analysis of total hCGβ in urine rather than in serum by immunomagnetic reduction (IMR) assay as an alternative method to diagnose an ectopic pregnancy. We established a standard calibration curve of IMR intensity against total hCGβ concentration based on standard hCGβ samples, and used an IMR assay to detect total hCGβ concentrations in the urine of pregnant women with lower abdominal pain and/or vaginal bleeding. The final diagnosis of ectopic pregnancy was based on ultrasound scans, operative findings, and pathology reports. In this prospective study, ten clinical samples were used to analyze the relationship of total hCGβ IMR signals between urine and serum. Furthermore, 20 clinical samples were used to analyze the...

Research paper thumbnail of Concordant myelomeningocele in dizygotic twins conceived by intracytoplasmic sperm injection, in vitro fertilization, and embryo transfer

Taiwanese journal of obstetrics & gynecology, 2013

Research paper thumbnail of Embryonic early-cleavage rate is decreased with aging in GnRH agonist but not inantagonist protocols

Journal of assisted reproduction and genetics, Jan 20, 2015

The aim of this study was to evaluate the correlation between embryonic early-cleavage status and... more The aim of this study was to evaluate the correlation between embryonic early-cleavage status and the age of patients receiving either a GnRH agonist long protocol or a GnRH antagonist protocol. This retrospective study included 534 patients undergoing a fresh cycle of oocyte retrieval and day-3 embryo transfer. Of the 534 patients treated, 331 received a GnRH agonist long stimulation protocol (GnRH agonist group) for ovarian stimulation and 203 patients received a GnRH antagonist protocol (GnRH antagonist group). By logistic regression analysis, the rate of embryonic early-cleavage was significantly decreased with increasing age of women in the agonist…

Research paper thumbnail of The safety and efficacy of microwave endometrial ablation after endometrial curettage without hormonal pretreatment

Taiwanese journal of obstetrics & gynecology, 2007

To evaluate the safety and efficacy of a microwave endometrial ablation (MEA) procedure to treat ... more To evaluate the safety and efficacy of a microwave endometrial ablation (MEA) procedure to treat patients with menorrhagia after endometrial curettage but without medical pretreatment. From February to September 2001, 19 eligible and consenting premenopausal women with menorrhagia underwent endometrial curettage, immediately followed by MEA. No medical pretreatment with gonadotropin-releasing hormone (GnRH) analogues or danazol was given for endometrial preparation. The severity of menorrhagia was assessed using the menstrual score. The patients were followed up for at least 3 years. Of the 19 women, 17 had completed at least 3 years' follow-up. Fourteen patients (82%) were satisfied with the posttreatment menstrual status, comparable with those patients receiving medical pretreatment of other studies (78-94%). The mean menstrual score was 11.8 before treatment and 1.95, 2.25, 2.2, 2.1 and 2.1 at 3, 6, 12, 24 and 36 months posttreatment, respectively. Of the 12 patients who comp...

Research paper thumbnail of The efficiency of conventional microscopic selection is comparable to the hyaluronic acid binding method in selecting spermatozoa for male infertility patients

Taiwanese journal of obstetrics & gynecology, 2015

To evaluate if hyaluronic acid (HA)-bound spermatozoa surpassed conventional microscopy-selected ... more To evaluate if hyaluronic acid (HA)-bound spermatozoa surpassed conventional microscopy-selected spermatozoa in the status of sperm DNA integrity by acridine orange (AO) fluorescence staining. Spermatozoa obtained from couples with indication for the intracytoplasmic sperm injection (ICSI) procedure due to male infertility (n = 34) and control males with normal sperm parameters (n = 12) were analyzed using AO fluorescence staining after density-gradient centrifugation (DGC), polyvinylpyrrolidone (PVP)-microscopic selection, and HA-binding selection to determine sperm DNA integrity. Percentages of DNA intact spermatozoa with green fluorescence were significantly higher in both PVP-microscopic selected spermatozoa (82.1 ± 24.0%) and HA-bound spermatozoa (83.9 ± 21.1%) than in spermatozoa prepared by DGC (66.8 ± 24.0%). However, there was no significant difference between the PVP-sperm and HA-sperm groups. When the percentage of green fluorescent spermatozoa prepared by DGC fell initia...

Research paper thumbnail of Embryonic Dormancy Phenomenon in Obstructed Healthy Mouse Fallopian Tubes

A mouse model of unilateral oviductal obstruction was designed to study whether healthy oviducts ... more A mouse model of unilateral oviductal obstruction was designed to study whether healthy oviducts can support embryo development in an advanced stage toward blastocyst hatching and implantation when the embryos in the contralateral side normally move into the uterine cavity. Methods: The oviducts of 80 female ICR mice (aged 5-8 weeks) were ligated unilaterally 12-40 hr postcoitus. The ligated oviducts were isolated from day 4 to 19.5 postcoitus. Embryos within the ligated oviducts were then flushed out to record the developmental stage and compared with the conceptuses in the contralateral uterine horns with unligated oviducts. Embryos recovered from ligated oviducts were then cultured in vitro to observe their potential for further development. Results: In 33 mice, 53.4% (163/305) and 86.1% (241/280) of the morphologically normal blastocysts had hatched from the zona pellucida within the obstructed tube and contralateral uterine horns, respectively, on the 5th day postcoitus. The data demonstrated that the hatching process could take place within the obstructed fallopian tube, but the timing was delayed. From 5.5 to 19.5 days postcoitus, a total of 362 implanted embryos were obtained in unligated control

Research paper thumbnail of Mosaic Trisomy 7 at Amniocentesis: Prenatal Diagnosis and Molecular Genetic Analyses

Taiwanese Journal of Obstetrics and Gynecology, 2010

Objective: To present prenatal diagnosis and molecular genetic analyses of mosaic trisomy 7. Mate... more Objective: To present prenatal diagnosis and molecular genetic analyses of mosaic trisomy 7. Materials, Methods and Results: A 38-year-old primigravid woman underwent amniocentesis at 19 weeks of gestation because of her advanced maternal age. Amniocentesis revealed a karyotype of 47,XY,+7[26]/46, XY[16]. Repeated amniocentesis at 21 weeks of gestation revealed a karyotype of 47,XY,+7[20]/46,XY[17]. Simultaneous cordocentesis revealed a karyotype of 46,XY in 100/100 cultured lymphocytes. Polymorphic DNA marker analyses of uncultured amniocytes and cord blood revealed a diallelic pattern with seemingly equal biparental inheritance of chromosome 7. Repeated cordocentesis and chorionic villus sampling at 23 weeks of gestation revealed a karyotype of 47,XY,+7[2]/46,XY[66] in cord blood and a karyotype of 47,XY,+7 in 24/24 cultured chorionic villi cells. Level II ultrasonography was normal. At 40 weeks of gestation, a 2,708 g normal male baby was delivered. The peripheral blood had a karyotype of 46,XY in 100/100 lymphocytes. Molecular analyses of placenta, urine, buccal swab, and peripheral blood revealed a diallelic pattern and seemingly equal biparental inheritance of chromosome 7 in all tissues. At 3 months of age, he manifested hypopigmented skin and inguinal hernia, but showed normal growth and mental development. Fluorescence in situ hybridization analysis of inguinal hernia sac tissue revealed that 19/100 (19%) of nuclei had three chromosome 7 signals. Conclusion: Mosaic trisomy 7 at amniocentesis may be derived from a cell culture artifact from an undetected low level of trisomy 7 mosaicism in uncultured amniocytes, and can be associated with favorable fetal outcome if the blood has a normal karyotype or a very low level of mosaicism and if uniparental disomy for chromosome 7 is excluded.

Research paper thumbnail of Clinical application of immunomagnetic reduction for quantitative measurement of insulin-like growth factor binding protein-1 in the prediction of pregnant women with preterm premature rupture of membranes

Clinica Chimica Acta, 2015

Background: The most important factor for a successful pregnancy after in vitro fertilization is ... more Background: The most important factor for a successful pregnancy after in vitro fertilization is embryo quality. The aim of this study was to explore the possibility that using the immunomagnetic reduction (IMR) assay to quantitatively measure β-subunit of human chorionic gonadotropin (β-hCG) in blastocyst culture media to differentiate embryo quality. Methods: This was a prospective case-control study including 28 samples of blastocyst culture media. We used single-step blastocyst culture and IMR assay to analyze β-hCG concentrations in culture media. We also explored the relationship between IMR signals of β-hCG and morphological grading of blastocysts. Results: β-hCG concentration-dependent IMR signals were highly correlated with blastocyst morphological quality (Spearman correlation coefficient: 0.731). Receiver-operating characteristic curve analysis showed a cutoff IMR value to differentiate embryo quality of 0.873%, with an area under the curve of 0.947, sensitivity of 0.882 and specificity of 0.818. Furthermore, subanalysis also revealed a positive correlation between β-hCG concentration-dependent IMR signals and trophectoderm grading, with a Spearman correlation coefficient of 0.576. Conclusions: An IMR assay can quantitatively measure β-hCG in blastocyst culture media, and may be a potential clinical tool to assist in the assessment of good blastocyst quality before embryo transfer.

Research paper thumbnail of Differential expression and distribution of alternatively spliced transcripts of PDGF-A and of PDGF receptor-alpha in mouse reproductive tissues

Life sciences, Jan 23, 2005

Platelet-derived growth factor-A (PDGF-A) affects cellular activities such as proliferation, diff... more Platelet-derived growth factor-A (PDGF-A) affects cellular activities such as proliferation, differentiation, and development by way of paracrine or autocrine interaction with PDGF-A receptor alpha (PDGFR-alpha). Two forms of alternatively spliced PDGF-A mRNA, a long and a short isoform, have been found in several mammalian species. Expression of PDGF-A and its cognate receptor PDGFR-alpha has been well studied in various tissues. However, these investigations did not distinguish between the individual isoforms of PDGF-A. In the present investigation, we identified the differential cellular expression patterns of the two isoforms of PDGF-A and of PDGFR-alpha in mouse reproductive tissues by using laser capture microdissection coupled with reverse transcriptase polymerase chain reaction. The long PDGF-A mRNA isoform was primarily detected in the epithelium, while the short isoform was ubiquitously distributed in epithelium, stroma, and muscle cells, although it was still more promine...

Research paper thumbnail of Relationship of follicular size to the development of intracytoplasmic sperm injection-derived human embryos

Taiwanese journal of obstetrics & gynecology, 2010

To compare the embryonic development of oocytes obtained from follicles of different sizes. Oocyt... more To compare the embryonic development of oocytes obtained from follicles of different sizes. Oocytes (n = 819) were retrieved from women at 40 years of age or younger during 86 in vitro fertilization cycles and categorized as small, medium, or large based on the estimated volume of follicular fluid at the time of retrieval. The rates of good quality embryos from the large, medium, and small groups on days 2 and 3 were 76.85% and 66.20%, 74.00% and 61.33%, and 69.81% and 58.49%, respectively. There were no significant differences in the rates of good quality embryos between the three follicular volume groups. Even though fewer oocytes completed maturation in the small follicle group than in the other two groups, the quality of the embryos in all three groups was the same on days 2 and 3. These findings suggest that follicles of all sizes should be aspirated during the intracytoplasmic sperm injection cycle as follicles of every size were a good source of embryos.

Research paper thumbnail of Comparison of day 2 tubal embryo transfer with day 3 embryo transfer into the uterus of intracytoplasmic sperm injection derived human embryos

Taiwanese journal of obstetrics & gynecology, 2006

To compare the results of embryo transfer into the uterus (ET) performed on day 3 vs. tubal embry... more To compare the results of embryo transfer into the uterus (ET) performed on day 3 vs. tubal embryo transfer (TET) on day 2 in couples diagnosed with male factor infertility. We collected data from 34 and 110 patients who underwent intracytoplasmic sperm injection (ICSI) followed by either TET on day 2 or ET on day 3 (January 2001-June 2005), respectively. All couples were diagnosed with male factor infertility; there were no tubal or uterine factors, diminished ovarian reserve, pelvic adhesions, history of polycystic ovary syndrome or previous ovarian surgery. The clinical pregnancy rates of day 2 TET and day 3 ET following ICSI were 64.71% and 57.27% (p > 0.05), respectively. The corresponding implantation rates were 35.47% and 29.58% (p > 0.05), respectively. The implantation and clinical pregnancy rates of ET on the 3rd day following ICSI were similar to the rates obtained with TET 2 days after ICSI. Therefore, ET performed 3 days after ICSI should be the first choice for c...

Research paper thumbnail of Early-cleavage is a reliable predictor for embryo implantation in the GnRH agonist protocols but not in the GnRH antagonist protocols

Reproductive Biology and Endocrinology, 2009

Background To test if early-cleavage was a strong predictor of pregnancy in patients receiving ei... more Background To test if early-cleavage was a strong predictor of pregnancy in patients receiving either a GnRH agonist long protocol or a GnRH antagonist protocol for in-vitro fertilization treatment (IVF) and intracytoplasmic sperm injection (ICSI). Methods This retrospective study included 534 patients undergoing a fresh cycle of oocyte retrieval and the day-3 embryo transfer (from 22 to 46 years old). Of the 534 patients treated, 331 received a GnRH agonist long stimulation protocol (GnRH agonist group) for ovarian stimulation and 203 patients received a GnRH antagonist protocol (GnRH antagonist group). In each group, patients who had at least one early-cleavage embryo transferred were designated as the 'early-cleavage' subgroup. Patients who had no early-cleavage embryos transferred were designated as the 'late-cleavage' subgroup. Results The early cleavage rate was significantly lower in the GnRH antagonist group compared with that in the GnRH agonist group (IVF c...

Research paper thumbnail of Prenatal identification of fetal overgrowth, abdominal wall defect, and neural tube defect in pregnancies achieved by assisted reproductive technology

Research paper thumbnail of Involvement of the Serine Protease Inhibitor, SERPINE2, and the Urokinase Plasminogen Activator in Cumulus Expansion and Oocyte Maturation

PLoS ONE, 2013

The serpin peptidase inhibitor, clade E, member 2 (SERPINE2) inhibits urokinase-type plasminogen ... more The serpin peptidase inhibitor, clade E, member 2 (SERPINE2) inhibits urokinase-type plasminogen activator (PLAU) and tissue-type plasminogen activator. Higher SERPINE2 expression levels were detected in cumulus cells of human immature oocytes than in those of mature oocytes. The objective of this study was to evaluate whether high SERPINE2 levels in cumulus cells are associated with oocyte immaturity. Using the mouse cumulus-oocyte complex as an experimental model, the effects of elimination and overexpression of SERPINE2 in cumulus cells on cumulus expansion and oocyte maturation were assayed by in vitro maturation. Serpine2 and PLAU transcripts were the most highly expressed serpins and plasminogen activators, respectively. Their expression was coordinately regulated in cumulus cells during gonadotropin-induced oocyte maturation. Silencing of Serpine2 expression using small interfering RNAs or blockage of SERPINE2 protein using a specific antibody had no effect on oocyte maturation. However, overexpression of Serpine2 or exogenous supplementation with high levels of SERPINE2 impaired cumulus expansion and oocyte maturation, probably by decreasing hyaluronan synthase 2 (Has2) and versican (Vcan) mRNA expression. Amiloride, a specific PLAU inhibitor, also suppressed these processes. PLAU supplementation of the oocyte in vitro maturation medium caused earlier and more extensive expansion of cumulus cells and oocyte maturation that may be mediated by increased Has2 mRNA expression. However, these effects were neutralized by coincubation with SERPINE2 or amiloride and PLAU. In conclusion, SERPINE2 and PLAU are involved in cumulus expansion and oocyte maturation. High SERPINE2 levels impair these processes, probably by decreasing cumulus matrix gene expression as well as reducing cumulus hyaluronan contents and inhibiting PLAU activity. These findings may explain why cumulus cells surrounding immature human oocytes express high SERPINE2 levels.