Characteristics of amniotic mesenchymal stromal cells derived from term and preterm labor.

OBJECTIVE: Amniotic mesenchymal stromal cells (AMSCs) can be obtained from the mesoderm of human amniotic membrane. AMSCs derived from term baby show increased expression of genes associated with apoptosis and senescence. The objective of this study was to examine gene expression profiles of AMSCs derived from preterm (preterm AMSCs) and term labors (term AMSCs) and analyze common and different mechanisms. MATERIALS AND METHODS: We isolated and cultured AMSCs from 43 placentas: 27 from term labor and 16 from preterm labor. Microarray analysis and gene network analysis were performed to compare gene expression profile (GEP) of preterm (n = 6) with term AMSCs (n = 10). Senescence-associated gene (CDKN2A and CDKN2B) expression was also measured by reverse transcription quantitative PCR. RESULTS: GEP demonstrated that preterm AMSCs showed upregulation of nicotinamide adenine dinucleotide biosynthetic process and downregulation of extracellular matrix, cholesterol import and transport, lipid storage, and maintenance of location. CDKN2A and CDKN2B genes showed similar expression levels between term and preterm AMSCs. CDKN2A gene expression was correlated with CDKN2B expression and population doubling time. Compared to term AMSCs, preterm AMSCs showed significantly different expression of genes associated with inflammatory response which could be one of the major players in labor events. CONCLUSION: Increased CDKN2A expression in AMSCs is associated with placental membrane aging which participates in both preterm and term labor. To the best of our knowledge, this is the first report to demonstrate the association of AMSCs with labor.

Optimizing the Diagnostic Strategy to Identify Genetic Abnormalities in Miscarriage.

INTRODUCTION: The single most common cause of miscarriage is genetic abnormality. OBJECTIVE: We conducted a prospective cohort study to compare the performance of conventional karyotyping and chromosomal microarray analysis (CMA) using array comparative genomic hybridization (array-CGH) and single nucleotide polymorphism array (SNP-array) to identify genetic abnormalities in miscarriage specimens. METHODS: A total of 63 miscarriage specimens were included. Conventional karyotyping, array-CGH, and SNP-array were performed and the results compared. RESULTS: Genetic abnormalities were detected in 31 cases (49.2%) by at least one testing modality. Single autosomal trisomy was the most common defect (71.0%), followed by polyploidy (16.1%), multiple aneuploidy (9.7%), and monosomy X (3.2%). Mosaicisms were identified in four cases and confirmed by fluorescence in situ hybridization (FISH) using appropriate probes. SNP-array had a higher detection rate of genetic abnormalities than array-CGH (93.5 vs. 77.4%), and conventional karyotyping had the lowest detection rate (76.0%). SNP-array enabled the detection of all types of genetic abnormalities, including polyploidy. CONCLUSIONS: Although conventional karyotyping and FISH are still needed, SNP-array represents the first choice for miscarriage because the technique showed excellent performance in the detection of genetic abnormalities and minimized the probability of testing failure as well as time, costs, and labor.

Anomalous Connection of Umbilical Vessels to the Left Ventricle: Case Report.

The anomalous connection of umbilical vessels to the heart is rare and has not yet been reported in the international scientific literature. Herein, we report the case of a newborn who was diagnosed with an anomalous connection of the umbilical vessels to the left ventricle. These anomalous vessels were functionally open for 2 weeks, and cellulitis was present in the area of the blood vessels connected to the skin. We performed division of these abnormal vessels and removal of the skin lesion.

Increased urinary neutrophil gelatinase-associated lipocalin in very-low-birth-weight infants with oliguria and normal serum creatinine.

BACKGROUND: In infants, oliguria is defined as a urine output of <1.5 mL/kg/h. The aim of our study was to assess the impact of oliguria on urinary neutrophil gelatinase-associated lipocalin (NGAL) and serum cystatin C (CysC) levels in very-low-birth-weight infants (VLBWIs) with a normal serum creatinine (Cr) level. METHODS: Fifty-seven VLBWIs were enrolled in the study. Urinary NGAL, serum CysC and Cr levels and urinary NGAL/Cr ratios were measured. Infants with Apgar scores of >5 at 5 min and/or a serum Cr level of >1.5 mg/dL or those treated for patent ductus arteriosus were excluded. In case of antibiotic treatment, blood and urine samples were collected at ≥48 h after discontinuation of antibiotic treatment. RESULTS: There was a significant difference in gestational age between infants with oliguric episodes during hospitalization and those without, but not in birth weight, perinatal or postnatal factors. Gestational age was negatively correlated with urinary NGAL and serum CysC levels and urinary NGAL/Cr ratio (p < 0.05), whereas postnatal age was negatively correlated with serum Cr level and urinary NGAL/Cr ratio (p < 0.05). Of the 117 urine and blood samples collected, 25 (21.4%) were obtained from neonates with oliguric episodes. After adjusting for gestational age and postnatal age, comparison of samples collected in infants with and without oliguric episodes revealed significant differences in the mean level of urinary NGAL and in the urinary NGAL/Cr ratio, but not in mean serum CysC or serum Cr levels. The urinary NGAL level [area under the curve (AUC) 0.886, 95% confidence interval (CI) 0.814-0.937] and urinary NGAL/Cr ratio (AUC 0.853, 95% CI 0.775-0.911) showed significantly greater discrimination for oliguria than serum CysC (AUC 0.610, 95% CI: 0.515-0.699) or serum Cr (AUC 0.747, 95%CI 0.659-0.823) levels. CONCLUSIONS: Urinary NGAL level and urinary NGAL/Cr ratio were more sensitive markers for the presence of oliguria in VLBWIs with normal serum Cr levels than serum CysC level.

Comparison of Laparoscopic Cornual Resection and Cornuotomy for Interstitial Pregnancy.

STUDY OBJECTIVE: To compare the clinical efficacy and safety of laparoscopic cornuotomy and cornual resection in the treatment of interstitial pregnancy. DESIGN: Retrospective chart review between 2006 and 2014 (Canadian Task Force classification II-2). SETTING: Two academic tertiary care hospitals. PATIENTS: Seventy-five patients with interstitial pregnancy treated by laparoscopy. MEASUREMENT AND MAIN RESULTS: In the 75 patients, 53 who underwent cornual resection and 22 who underwent cornuotomy, we evaluated operating time, changes in hemoglobin levels after surgery, the rate of major complications, and the incidence of persistent interstitial pregnancy. The mean operating time was significantly shorter for cornuotomy than for cornual resection (59.36 ± 19.32 minutes vs. 77.11 ± 23.97 minutes, respectively). Changes in hemoglobin level after the operation, rates of major complications, and the incidence of persistent interstitial pregnancy were not significantly different in the 2 surgery groups. CONCLUSION: Laparoscopic cornuotomy yielded clinical results comparable to those of cornual resection. Laparoscopic cornuotomy may reduce the time of operation, and had the same incidence of persistent interstitial pregnancy as cornual resection.

Single-port laparoscopic debulking surgery of variant benign metastatic leiomyomatosis with simultaneous lymphatic spreading and intraperitoneal seeding.

Benign metastatic leiomyomatosis (BML) is a rare disease characterized by smooth muscle cell proliferation in extrauterine sites including the lung, abdomen, pelvis, and retroperitoneum. Depending on location, BML is classified as intravenous leiomyomatosis and diffuse peritoneal leiomyomatosis. Pathogenesis of BML can be iatrogenic after previous myomectomy or hysterectomy, hormonal, or coelomic metaplasia. Treatment options are observation, hormonal suppression, and/or surgical debulking via laparotomy or laparoscopy. Laparoscopic surgery is gaining in popularity in the gynecologic field compared to laparotomic surgery and single-port laparoscopy has the benefits of cosmesis and early tissue extraction by transumbilical morcellation. We report a 39-year-old woman with BML who underwent single-port laparoscopy debulking surgery.

[Proportions of cells expressing CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, or CD13,33+/CD34+ in the regenerating bone marrows during complete remission of acute leukemia or after bone marrow transplantation].

BACKGROUND: The hemopoietic stem cells increase in number during the regeneration after chemotherapy or bone marrow transplantation (BMT). Although the proportion of hemopoietic stem cells and their differentiation have been studied by immunophenotyping using the flow cytometry, no substantial research efforts have been directed toward the regenerating marrow. We attempted to discover the proportions of undifferentiated stem cells, committed stem cells, B cell precursors, and myeloid precursors in the regenerating bone marrows during complete remission (CR) and after engraftment of BMT. METHODS: Bone marrow samples from 82 patients with acute leukemia in CR and from 25 patients after BMT engraftment, along with 22 control samples, were used to find the numbers of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells in the large lymphocyte gate by flow cytometry. We cross-analyzed our results in terms of groups: CR, BMT, and initial diagnosis groups. We performed significance tests on age, relapse, chromosomal abnormalities, clinical outcomes, and initial immunophenotypes of the leukemic cells. RESULTS: The proportions of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells are more highly distributed in acute B-lymphoblastic leukemia than the normal group and also in the CR than the BMT group. CD19+/CD34+ cells were increased in the relapse group and CD38+/ CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells were increased in the group with chromosomal abnormality. The results were irrelevant to the initial immunophenotype of the leukemic blasts. CONCLUSIONS: The increases of the markers spanned too widely to apply one specific cutoff value to analyze them. They seemed to be the results of normal regeneration, irrelevant to relapse or initial immunophenotype of leukemic blasts.

Pioglitazone for treating polycystic ovary syndrome in non-obese women of reproductive age with different clinical presentations.

AIMS: To evaluate the effects of pioglitazone on menstruation and the metabolic parameters of non-obese women of reproductive age with polycystic ovary syndrome (PCOS) and compare the effects among different subgroups of PCOS patients with different clinical presentations. METHODS: Twenty-eight women of reproductive age with PCOS were recruited; 20 women finished the study. The women were divided into three groups according to clinical presentations: group A (n = 4) had chronic oligo- or anovulation with polycystic ovaries; group B (n = 5) had chronic oligo- or anovulation with hyperandrogenism; and group C (n = 11) had chronic oligo- or anovulation, hyperandrogenism and polycystic ovaries. Pioglitazone (15 mg/day) was given for 6 months and the therapeutic effects were evaluated. Menstrual cycle regularity and hormone levels (plasma luteinizing hormone (LH), follicle-stimulating hormone, total cholesterol, triglycerides, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol, glucose, insulin, C-peptide, free testosterone, homeostatic model assessment (HOMA)) were evaluated during and after pioglitazone treatment. RESULTS: Thirty percent and 45.5% of the patients showed improvement of menstrual cycle regularity immediately after completion of treatment and at 6 months after completion of pioglitazone treatment, respectively, although there were no statistical differences among the subgroups. There was a significant difference in the change of body mass index (BMI) throughout the study period among the subgroups (p = 0.008). The decrease in BMI was significantly higher in group B than in groups A and C at 3 months (p = 0.0381) and 6 months of treatment (p = 0.0054), as well as at 6 months after completion of treatment (p = 0.003). HDL-C concentrations increased throughout the period (p = 0.001) without a difference among the subgroups. LH levels decreased at 6 months of treatment and throughout the follow-up period (p = 0.0045), but this did not differ among subgroups. The free testosterone level decreased, but without significance. There was no statistical improvement in any of the parameters of insulin resistance, but baseline free testosterone levels were related to the improvement of the HOMA insulin sensitivity score (p = 0.0009). Patients with more than a 50% decrease of their HOMA insulin resistance (HOMA-IR) score showed higher baseline free testosterone levels (hyperandrogenic groups B and C) than did the patients with less than 50% HOMA-IR score improvement or those patients with an increased HOMA-IR score. CONCLUSIONS: These preliminary results suggest that pioglitazone treatment for non-obese PCOS women of reproductive age may be effective to help the resumption of the menstrual cycle, and the patients showed a different response pattern according to their baseline free testosterone levels.

The incidence and characteristics of uterine bleeding during postoperative GnRH agonist treatment combined with tibolone add-back therapy in endometriosis patients of reproductive age.

OBJECTIVES: This study was undertaken to evaluate the incidence and the factors associated with uterine bleeding while taking GnRH agonist treatment combined with tibolone add-back therapy in endometriosis patients of reproductive age. STUDY DESIGN: The medical records of 188 endometriosis (stages III-IV) patients of reproductive age receiving postoperative GnRH agonist treatment combined with tibolone add-back therapy for 6 months were reviewed. Clinical features were analyzed and compared between the two groups: Group A, patients without an episode of uterine bleeding (n=137) and Group B, patients that had experienced uterine bleeding (n=51). RESULTS: Demographic profiles were not different in the two groups. The incidence of uterine bleeding was 27.1% and irregular spotting was the most frequent bleeding pattern. The proportion of patients who received ovarian surgery during pelviscopy was higher in Group B (p<0.05), but other clinical characteristics were not different between two groups. CONCLUSIONS: The incidence of uterine bleeding was 27.1%. And, the proportion of patients who received ovarian surgery was higher in patients with uterine bleeding.

Subserosal pregnancy in a previous myomectomy site: a variant of intramural pregnancy.

A 35-year-old woman with a history of myomectomy underwent in vitro fertilization and became pregnant. Transvaginal ultrasound revealed a gestational sac within the subserosal area of the posterior uterine wall. The patient was treated successfully with conservative surgery, and the pathologic evaluation of the excised mass demonstrated chorionic villi involving myometrium. Early in a subsequent pregnancy, placental invasion through the sinus tract was detected. However, the pregnancy outcome was uneventful. This constitutes the first report of subserosal implantation in the uterine body. Our findings suggest that the probable pathogenesis of this rare variant of intramural pregnancy is implantation through a sinus tract made during a previous uterine surgery.

Protective effect of vascular endothelial growth factor (VEGF) in frozen-thawed granulosa cells is mediated by inhibition of apoptosis.

OBJECTIVES: This study was conducted to evaluate the involvement of apoptosis in the freeze-thaw process and to investigate the anti-apoptotic effect of vascular endothelial growth factor (VEGF) in the frozen-thawed granulosa cells. STUDY DESIGN: Isolated rat granulosa cells were cultured, frozen-thawed, and were cultured for 24h. Cell viabilities (by Trypan blue exclusion test) and apoptotic patterns (by Annexin-V/propidium iodide (PI) Double-Staining) were determined at each step. Apoptotic cell death was confirmed by following DNA degradation and caspase-3 activity. RESULTS: After freeze-thaw process and 24h of culture, reductions in the cellular viabilities and increases in the number of cells containing degraded DNA were lower in the VEGF pretreated group than in the control group (p<0.05). In the VEGF pretreated group, increases in the proportions of late apoptotic cells [Annexin-V (+)/PI (+)] were significantly lower and caspase-3 expression was prevented immediate after thawing (p<0.05). Furthermore, increases in the proportions of early apoptotic cells [Annexin-V (+)/PI (-)] and reductions in the proportions of viable cells [Annexin-V (-)/PI (-)] were significantly lower in the VEGF pretreated group after culture for 24h (p<0.05). Of the different doses of VEGF pretreated, 50ng/ml was found to be most effective with respect to protecting frozen-thawed granulosa cells from cryoinjury. CONCLUSION: Granulosa cell damage induced by cryopreservation is mediated, at least in part, by an apoptotic process. Our preliminary results suggest that VEGF treatment before freeze-thaw process reduces rat ovarian granulosa cell damage by inhibiting apoptosis.

The regulators of VEGF expression in mouse ovaries.

The objectives of this study were to explore whether ovarian vascular endothelial growth factor (VEGF) expression in mice can be regulated by IL-6 (interleukin-6), angiotensin II, FSH, and hCG; and to test whether the mouse ovarian VEGF expression can result in angiogenesis. The ICR mice were sacrificed, and their ovaries were recovered. Recovered ovaries were treated with IL-6, angiotensin II, FSH, and hCG separately and incubated for 24 hours in alpha-MEM. Expression of mRNA and protein of VEGF were assessed by RT-PCR and immunohistochemistry. The resulting angiogenesis was evaluated through immunohistochemical analysis for CD34. Treatment of mice ovaries with IL-6, FSH, and hCG resulted in a significant increase of VEGF mRNA, and IL-6 was the most potent inducer of VEGF. IL-6 and FSH resulted in increased neovascularization in the follicular phase of mouse ovaries. In contrast, angiotensin II could not increase VEGF expression or neovascularization. We documented an in vitro increase in VEGF expression by IL-6, FSH, and hCG; and reaffirmed that the proliferative response of murine ovarian endothelial cells paralleled an increase of VEGF expression.

Myelodysplastic syndrome with erythroid aplasia following pure red cell aplasia.

Myelodysplastic syndrome (MDS) with erythroid aplasia is a very rare disorder that has not been clearly defined. We experienced a case of pure red cell aplasia (PRCA), which evolved to MDS with erythroid aplasia. A 59-year-old male with transfusion-dependent PRCA was referred to our hospital for an evaluation of newly developed thrombocytopenia. Two years ago, PRCA was diagnosed by the laboratory findings and a bone marrow examination, which showed no evidence of any myelodysplastic features and thymoma. Upon admission, the bone marrow findings showed marked hypercellularity. with numerous dysplastic features in the three lineages including erythroid hypoplasia. These findings were compatible with a diagnosis of MDS with red cell aplasia. It is very interesting that the PRCA evolved to MDS with red cell aplasia, which strongly suggests an autoimmune mechanism for the development of MDS.

Early and late hormonal responses to the microdose gonadotropin-releasing hormone agonist in normal menstruating women.

OBJECTIVE: To assess the effect of a microdose gonadotropin-releasing hormone (GnRH) agonist on the LH, FSH, and E2 secretion in normal menstruating women. DESIGN: Prospective study. SETTING: Tertiary teaching hospital. PATIENT(S): Five normal menstruating women. INTERVENTION(S): Five microg of triptorelin was injected daily in 5 women for 7 days beginning from the cycle day 3. In the next cycle, the same amount of triptorelin was injected into the same women daily for 3 days. MAIN OUTCOME MEASURE(S): Serial serum FSH, LH, and E2 levels. RESULT(S): The FSH levels peaked (27.53 +/- 6.34 IU/L) after 5 hours, and the LH levels peaked (34.35 +/- 7.81 IU/L) by 4 hours. The increased gonadotropin levels persisted even after the second and third day of the GnRH-agonist injections, although the peak levels were not as high as observed with the first injection (19.56 IU/L in the second day, 9.15 IU/L in the third day for FSH; 32.18 IU/L in the second day, 13.59 IU/L in the third day for LH). The down-regulation of gonadotropins was established in 4 days. When the GnRH-agonist was administered for 7 days, the E2 level began to increase 6 days after the last injection. When the GnRH-agonist was administered for 3 days, the E2 level began to increase 3 days after the last injection. CONCLUSION(S): Pituitary down-regulation could be achieved even with a microdose of GnRH agonist. The increased level of gonadotropins persisted for 3 days at this dose. The duration of the down-regulation was influenced by the duration of GnRH-agonist administration.

Intrapleural chemotherapy with cisplatin and cytarabine in the management of malignant pleural effusion.

PURPOSE: The purpose of this study was to evaluate the efficacy of intrapleural chemotherapy (IPC) with cisplatin and cytarabine in the management of malignant pleural effusion (MPE) from non-small-cell lung cancer (NSCLC). MATERIALS AND METHODS: A prospective analysis was carried out on 40 patients with pathologically proven MPE from NSCLC who had received IPC. A single dose of cisplatin 100 mg/m(2) plus cytarabine 1200 mg/m(2) in 250 ml normal saline was instilled into the pleural space via a chest tube and drained 4 hours later. Patients were evaluated for toxicities and responses at 1, 2, & 3 weeks and then at monthly intervals if possible. Systemic chemotherapy was administered, if the patient agreed to receive it, after achieving complete control (CC) of MPE. RESULTS: The median duration of chest tube insertion for drainage was 7 (3 approximately 32) days. Among the assessable 37 patients, CC and partial control (PC) were 32 (86.5%) and 4 (10.8%) patients, respectively (overall response rate 97.3%). The median duration of response was 12 months (2 approximately 23) and there were only two relapses of IPC after achieving CC. Among the 35 patients who were assessable until they died, 28 patients (80.0%) maintained CC until the last follow-up. There was only one toxic death and the toxicities of IPC, versus the results obtained, were deemed acceptable. CONCLUSION: The procedures were tolerable to the patients and chemotherapy-induced complications were at an acceptable level. The outcome of this trial indicates that IPC has a superior and long lasting treatment response in the management of patients with MPE from NSCLC.