Hsien-An Pan

The Expression Level of Septin12 Is Critical for Spermiogenesis

Septins belong to a family of polymerizing GTP-binding proteins that are required for many cellular functions, such as membrane compartmentalization, vesicular trafficking, mitosis, and cytoskeletal remodeling. One family member, septin12, is expressed specifically in the testis. In this study, we found septin12 expressed in multiple subcellular compartments during terminal differentiation of mouse germ cells. In humans, the testicular tissues of men with either hypospermatogenesis or maturation arrest had lower levels of SEPTIN12 transcripts than normal men. In addition, increased numbers of spermatozoa with abnormal head, neck, and tail morphologies lacked SEPT12 immunostaining signals, as compared with normal spermatozoa. To elucidate the role of septin12, we generated 129 embryonic stem cells containing a septin12 mutant allele with a deletion in the exons that encode the N-terminal GTP-binding domain. Most chimeras derived from the targeted embryonic stem cells were infertile, and the few fertile chimeras only produced offspring with a C57BL/6 background. Semen analysis of the infertile chimeras showed a decreased sperm count, decreased sperm motility, and spermatozoa with defects involving all subcellular compartments. The testicular phenotypes included maturation arrest of germ cells at the spermatid stage, sloughing of round spermatids, and increased apoptosis of germ cells. Electron microscopic examination of spermatozoa showed misshapen nuclei, disorganized mitochondria, and broken acrosomes. Our data indicate that Septin12 expression levels are critical for mammalian spermiogenesis.

Ovarian stroma flow intensity decreases by age: a three-dimensional power doppler ultrasonographic study

This was a prospective comparative clinical study to test the hypothesis that the flow intensity of the ovarian stroma decreases in the order of the aging process. A total of 100 consecutive women who came to our outpatient clinic for Pap smear examination were recruited. They were divided into three groups. The premenopause group (58 women) had menstruated within the last 3 months and had normal ovaries (no polycystic ovary or any pathologic cyst or mass), as demonstrated on the baseline ultrasound (US) examination. The perimenopause group (20 women) had last menstruated between 3 and 12 months earlier and had normal ovaries. The postmenopause group (22 women) had had no menstrual cycle within the last 12 months and had normal ovaries. Three-dimensional power Doppler US was applied to quantify the blood flow and vascularization within the stroma of the bilateral ovaries. The results showed that the E2 level decreased in the order of: premenopause (mean +/- SD; 40.88 +/- 40.65 pg/mL), perimenopause (22.00 +/- 13.61 pg/mL), then postmenopause (17.25 +/- 16.40 pg/mL). The vascularization index (VI) (6.95 +/- 8.35; 1.11 +/- 0.93; 0.53 +/- 1.75; respectively), flow index (FI) (15.98 +/- 7.59; 12.00 +/- 3.86; 5.18 +/- 5.31; respectively) and vascularization-flow index (VFI) (1.25 +/- 1.59; 0.18 +/- 0.15; 0.09 +/- 0.32; respectively) all decreased significantly in the order of premenopause, perimenopause, then postmenopause. To the best of our knowledge, this is the first study using 3-D power Doppler sonography that proves that the flow intensity decreases along with the aging process in the ovarian stroma.

Three‐dimensional power Doppler imaging of ovarian stromal blood flow in women with endometriosis undergoing in vitro fertilization

The aims of this retrospective study were to investigate whether the quantification of ovarian stromal blood flow and/or leptin concentration are predictive of in vitro fertilization (IVF) outcomes in women after laparoscopic ovarian cystectomy for large endometriomas.

Upregulation of Steroidogenic Enzymes and Ovarian 17β-Estradiol in Human Granulosa-Lutein Cells by Cordyceps sinensis Mycelium

Abstract There is increasing evidence that 17β-estradiol (E2) directly influences the quality of maturing oocytes and thus the outcome of assisted reproduction treatment. Although Cordyceps sinensis (CS) mycelium, a Chinese herbal medicine, is believed to enhance libido and fertility in both sexes, the mechanism of its effect in women has not been determined. The aim of the present study was to evaluate the effects of CS on steroidogenic enzyme expression and E2 biosynthesis in human granulosa-lutein cells (GLC). We found that CS induced E2 production by GLC in a dose- and time-dependent manner and that a 3-h treatment with CS induced increased levels of mRNAs coding for the P450 side chain cleavage enzyme (P450scc), 3β-hydroxysteroid dehydrogenase (3β-HSD), and aromatase. Western blot analysis demonstrated that, after treatment with CS for 3 h, protein levels of steroidogenic acute regulatory protein (StAR) and aromatase were upregulated while P450scc and 3β-HSD levels showed no substantial change. New protein synthesis was required for CS-induced E2 production because it was abrogated by cycloheximide pretreatment. Addition of 22(R)-hydroxycholesterol, thus bypassing the need for StAR protein, did not induce as much E2 production as CS treatment, indicating that upregulation of StAR protein was not the only factor contributing to CS-induced steroidogenesis. Cotreatment of GLCs with CS and aminoglutethimide, an aromatase inhibitor, completely abolished CS-induced E2 production. In conclusion, treatment of GLCs with CS results in increased E2 production due, at least in part, to increased StAR and aromatase expression. These data may help in the development of treatment regimens to improve the success rate of in vitro fertilization.

Quantification of ovarian Doppler signal in hyperresponders during in vitro fertilization treatment using three-dimensional power Doppler ultrasonography

To test the hypothesis that the increased ovarian sensitivity to gonadotropins observed in women embarking on an in vitro fertilization (IVF) treatment may be due to changes in ovarian stromal blood flow, we undertook this prospective comparative clinical study using three-dimensional (3-D) power Doppler ultrasound (US). The 3-D power Doppler ultrasonographic indexes were used to quantify ovarian stromal blood flow and vascularization in hyperresponders. A total of 58 patients undergoing an IVF cycle were recruited and divided into two groups, a hyperresponder group (n = 23) (peak estradiol > 3000 pg/mL or >/= 15 oocytes retrieved) and normal responders (n = 35), based on their response to a standard down-regulation protocol for controlled ovarian stimulation. During ovarian stimulation, on the day of human chorionic gonadotropin (HCG) administration, patients underwent hormonal (serum E2), ovarian volume and 3-D power Doppler (ovarian stroma flow) evaluation. The serum estradiol levels on the day of HCG administration, the number of oocytes retrieved and the ovarian volume were significantly higher in the hyperresponders than in the normal groups. The vascularization flow index (VFI), flow index (FI), and vascularization index (VI), were significantly higher (p < 0.05) in the hyperresponders (1.18 +/- 0.60, 50.23 +/- 2.81 and 2.27 +/- 1.08, respectively), compared to the women with a normal response (0.63 +/- 0.61, 43.19 +/- 7.81 and 1.25 +/- 1.18, respectively). Our study may help to explain the excessive response during gonadotropin administration in the hyperresponsive women.

Effects of Laparoscopic Ovarian Drilling on Young Adult Women with Polycystic Ovarian Syndrome

STUDY OBJECTIVE To assess changes in serum hormone levels and ovarian stromal blood flow after laparoscopic ovarian drilling (LOD) in young adult women with polycystic ovary syndrome (PCOS). DESIGN Prospective, nonrandomized study (Canadian Task Force classification II-1). SETTING Tertiary care, major teaching hospitals. PATIENTS Anovulatory young women with PCOS who were resistant to clomiphene citrate. INTERVENTION Laparoscopic ovarian drilling. MEASUREMENTS AND MAIN RESULTS To evaluate the endocrinological effects of LOD, serum leptin, insulin-like growth factor-1, estrone (E1), and estradiol were measured before and after ovarian drilling in the early follicular phase. Three-dimensional transabdominal power Doppler examinations were performed to determine the effects of LOD. Serum leptin was correlated with body mass index (BMI) before LOD. Levels of BMI, fasting blood sugar, and leptin were higher and LH, LH/FSH, and the sugar/insulin ratio were lower in the obese group. There were significant decreases in the free androgen index, and total testosterone, luteinizing hormone (LH), and LH/follicle-stimulating hormone (FSH) levels, and a significant increase in sex hormonebinding globulin (SHBG) concentration in the 3 months after the operation. The vascularization index and vascularization flow index of the intraovarian stroma significantly decreased after treatment. Reversed correlations between leptin and LH, LH/FSH, E1, thyroid-stimulating hormone, and SHBG were noted 3 months after the operation compared with levels obtained before the operation. CONCLUSIONS Treatment of young adult women with PCOS using LOD did not influence leptin levels but changed the ovarian stromal blood flow dynamics during short-term follow-up. The surgical procedure may be beneficial both to endocrine profiles and to intraovarian stromal flow in patients with PCOS.

SEPT12 deficiency causes sperm nucleus damage and developmental arrest of preimplantation embryos

Oocytes fertilized with spermatozoa obtained from Septin 12+/- chimeric mice failed to develop beyond the morula stage after IVF and intracytoplasmic sperm injection because of significant DNA defects in the spermatozoa. Given that SEPT12 is expressed at the edge of the sperm nucleus in both humans and mice, we hypothesized the vital roles of Septin 12 in sperm head shaping, nuclear DNA condensation, and early embryonic development.

Three-dimensional and power Doppler ultrasonography in infertility and reproductive endocrinology.

Three-dimensional (3D) ultrasound is an easy and noninvasive technique in the clinical practice of obstetrics and gynecology. It can be used in the analysis of morphologic anatomy and volume measurement and is highly reproducible. Power Doppler ultrasound can further depict and quantify the microcirculation of the target organ or the region of interest. The aim of this review is to report the present status and the development of 3D ultrasonography, as well as 3D power Doppler in the field of infertility and reproductive endocrinology. 3D ultrasound may help us investigate the functional and potential roles in various pathologic conditions of the female pelvis. Furthermore, 3D ultrasound may also provide substantial assistance in the clinical application of infertility and reproductive endocrinology.

Quantification of ovarian stromal Doppler signals in postmenopausal women receiving hormone replacement therapy.

ObjectiveTo investigate the effect of continuous-combined hormone replacement therapy (HRT) on ovarian circulation, we used three-dimensional power Doppler ultrasound to assess ovarian stromal blood flow in postmenopausal women. DesignForty healthy, postmenopausal women were enrolled in this prospective, controlled study. They were divided into two groups: the control group (20 patients with no HRT treatment) and the HRT group [initially 20 patients, but later 15 patients who completed the study, using continuous-combined conjugated equine estrogen (0.625 mg/day) plus medroxyprogesterone (5 mg/day)]. All patients underwent transvaginal three-dimensional power Doppler ultrasonography at the time of recruitment and 3 months later. The ovarian stromal flow indices, including vascularization-flow index (VFI), flow index (FI), and vascularization index (VI), were measured. Blood withdrawals for serum follicle-stimulating hormone and estrogen level testing were obtained before and after 3 months of treatment. ResultsAll the variables in the hormone profile showed significant difference (P < 0.05) after 3 months of treatment in the HRT group. Of interest, the three-dimensional power Doppler indices of ovarian stromal flow, VFI (0.13 ± 0.11 → 0.59 ± 0.49), FI (30.47 ± 12.06 → 38.41 ± 10.21), and VI (0.31 ± 0.27 → 1.12 ± 0.95) all showed significant increase (P < 0.05) after 3 months of HRT treatment. ConclusionsThere was a significant increase in ovarian stromal flow indices after 3 months of treatment in the HRT group, but not in the controls. Monitoring the ovarian flow changes by three-dimensional power Doppler may be of clinical importance when HRT is given.

Clinical suppression of precocious puberty with cetrorelix after failed treatment with GnRH agonist in a girl with gonadotrophin-independent precocious puberty

This report presents the case of a 7-year-old girl with gonadotrophin-independent precocious puberty treated with cetrorelix [gonadotrophin-releasing hormone (GnRH) antagonist] after poor response to GnRH agonist therapy was observed in the endocrinology outpatient clinic. Uterine and ovarian morphology returned to within the normal prepubertal range after GnRH antagonist was injected subcutaneously. Vaginal bleeding stopped completely. The effects of GnRH antagonist treatment were comparable to those of GnRH agonist. The potential advantage of GnRH antagonists would be a clinically significant direct effect on the ovary, if it exists, and GnRH antagonists should be available for use in such children.