TGF-ß and SHH Regulate Pluripotent Stem Cell Differentiation into Brain Microvascular Endothelial Cells in Generating an In Vitro Blood-Brain Barrier Model.

Blood-brain barrier (BBB) models are important tools for studying CNS drug delivery, brain development, and brain disease. In vitro BBB models have been obtained from animals and immortalized cell lines; however, brain microvascular endothelial cells (BMECs) derived from them have several limitations. Furthermore, obtaining mature brain microvascular endothelial-like cells (BME-like cells) from human pluripotent stem cells (hPSCs) with desirable properties for establishing BBB models has been challenging. Here, we developed an efficient method for differentiating hPSCs into BMECs that are amenable to the development and application of human BBB models. The established conditions provided an environment similar to that occurring during BBB differentiation in the presence of the co-differentiating neural cell population by the modulation of TGF-ß and SHH signaling. The developed BME-like cells showed well-organized tight junctions, appropriate expression of nutrient transporters, and polarized efflux transporter activity. In addition, BME-like cells responded to astrocytes, acquiring substantial barrier properties as measured by transendothelial electrical resistance. Moreover, the BME-like cells exhibited an immune quiescent property of BBB endothelial cells by decreasing the expression of adhesion molecules. Therefore, our novel cellular platform could be useful for drug screening and the development of brain-permeable pharmaceuticals.

IL-17A and Th17 Cells Contribute to Endometrial Cell Survival by Inhibiting Apoptosis and NK Cell Mediated Cytotoxicity of Endometrial Cells via ERK1/2 Pathway.

Immune status including the immune cells and cytokine profiles has been implicated in the development of endometriosis. In this study, we analyzed Th17 cells and IL-17A in peritoneal fluid (PF) and endometrial tissues of patients with (n=10) and without (n=26) endometriosis. Our study has shown increased Th17 cell population and IL-17A level in PF with endometriosis patients. To determine the roles of IL-17A and Th17 cells in the development of endometriosis, the effect of IL-17A, major cytokine of Th17, on endometrial cells isolated from endometriotic tissues was examined. Recombinant IL-17A promoted survival of endometrial cells accompanied by increased expression of anti-apoptotic genes, including Bcl-2 and MCL1, and the activation of ERK1/2 signaling. In addition, treatment of IL-17A to endometrial cells inhibited NK cell mediated cytotoxicity and induced HLA-G expression on endometrial cells. IL-17A also promoted migration of endometrial cells. Our data suggest that Th17 cells and IL-17A play critical roles in the development of endometriosis by promoting endometrial cell survival and conferring a resistance to NK cell cytotoxicity through the activation of ERK1/2 signaling. Targeting IL-17A has potential as a new strategy for the treatment of endometriosis.

Visceral obesity as a risk factor of incisional hernia after single-port laparoscopic gynecologic surgery.

OBJECTIVE: We aimed to evaluate associations between abdominal fat distribution (AFD) parameters and incisional hernia (IH) in patients who underwent transumbilical single-port laparoscopic surgery (SPLS) for gynecological disease. METHODS: Medical records of 2116 patients who underwent SPLS for gynecological disease at Daejeon St. Mary's Hospital between March 2014 and February 2021 were reviewed. Among 21 (1.0%) patients who developed IH requiring surgical treatment after SPLS, 18 had preoperative abdominopelvic computed tomography (CT) images. As a control group, we randomly selected 72 patients who did not develop IH and who had undergone preoperative abdominopelvic CT scan, matched to test patients by type of surgery. Total fat area (TFA), visceral fat area (VFA), subcutaneous fat area (SFA), visceral-to-subcutaneous fat ratio (VSR), and waist circumference (WC) were measured at the level of the third lumbar vertebral body on the preoperative abdominopelvic CT images, using National Institutes of Health (NIH) ImageJ version 1.53 k. RESULTS: Receiver operating curve analysis showed that VFA has the highest predictive value for IH among AFD parameters (AUC = 0.749, 95% CI 0.630-0.869, p < 0.001). Univariate analysis showed that age, BMI, hypertension, dyslipidemia, TFA, VFA, VSR and WC were significant factors for IH. In multivariate analysis, only high VFA was identified as an independent risk factor for IH (HR 6.18, 95% CI 1.13-33.87, p = 0.04), whereas BMI, TFA, SFA, VSR, and WC failed to show statistical significance. CONCLUSION: We could find high VFA as an independent risk factor of IH in patients who underwent SPLS for gynecologic disease.

Association of hormone therapy and changes of objective sleep quality in women of late menopausal transition with sleep disorder: a preliminary study.

OBJECTIVE: The aim of this study was to investigate changes in objective sleep quality with hormone therapy (HT) in women with late menopausal transition. METHODS: Healthy midlife women with sleep difficulty who received HT were included. Those undergoing late menopausal transition were screened. Sleep patterns and self-reported questionnaires were collected before and 10 weeks after starting HT. RESULTS: Ten women who met the criteria (age, 50.1 ± 2.8 years) showed higher sleep efficiency and shorter wakefulness after sleep onset (WASO) 10 weeks after starting HT. However, no significant change was found in objective sleep quality after adjustment for multiple comparisons: sleep efficiency, 84.2 ± 7.7 versus 88.2% ± 4.7%, P = 0.037, adjusted P = 0.259; WASO, 59.0 ± 27.2 minutes versus 41.4 ± 17.4 minutes, P = 0.020, adjusted P = 0.140; average duration per awakening, 2.9 ± 1.0 minutes versus 2.2 ± 0.5 minutes, P = 0.033, adjusted P = 0.231. A better score of subjective sleep quality in the Pittsburgh Sleep Quality Index was observed 10 weeks after starting HT (2.0 ± 0.0 vs 1.2 ± 0.4, P = 0.006, adjusted P = 0.042), but sensitivity analysis did not show consistent results after adjustment for multiple comparisons (2.0 ± 0.0 vs 1.1 ± 0.4, P = 0.020, adjusted P = 0.140). Total scores of the Insomnia Severity Index and Menopause Rating Scale were better 10 weeks after starting HT (Insomnia Severity Index, 14.7 ± 3.0 vs 9.1 ± 3.8, P = 0.010; Menopause Rating Scale, 29.0 ± 5.2 vs 21.6 ± 3.0, P = 0.009) with consistent results in sensitivity analyses. There was no difference in the Epworth Sleepiness Scale before and after HT (7.2 ± 1.7 vs 8.6 ± 4.5, P = 0.309). The change in each objective sleep quality variable before and after HT showed strong positive or negative correlations with the change in only a few items in subjective sleep quality. CONCLUSION: Women in the late menopausal transition period showed higher sleep efficiency and shorter WASO after HT; however, multiple comparisons showed no statistically significant difference in objective sleep quality between before and after HT.

Association between Serum Uric Acid Levels and Bone Mineral Density in Postmenopausal Women: A Cross-Sectional and Longitudinal Study.

BACKGROUND: Uric acid is one of natural antioxidants in human body. There have been several studies on the correlation between uric acid with oxidative stress and osteoporosis. However, the data are insufficient and results are controversial. In this regard, we determined the association between uric acid levels and bone mineral density (BMD) during the postmenopausal period. METHODS: We analyzed data from 328 postmenopausal women (mean age, 57.3 ± 6.5 years; mean serum uric acid level, 4.6 ± 1.0 mg/dL). The participants were divided into three groups based on tertiles of the serum uric acid level. The participants receiving hormone replacement therapy (HRT), bisphosphonates, or lipid-lowering agents were included. RESULTS: Blood urea nitrogen, serum creatinine, and serum triglyceride levels were significantly higher in the upper tertiles of uric acid levels. No significant difference was found in the mean uric acid levels between medication users and non-users. Each HRT regimen had a different mean serum uric acid level. A cross-sectional analysis showed no significant correlation between the serum uric acid levels and BMD in the spine and femoral neck (spine BMD: 1.050 ± 0.131, 1.060 ± 0.160, 1.084 ± 0.140, p = 0.22; femoral neck BMD: 0.837 ± 0.110, 0.849 ± 0.096, 0.863 ± 0.115, p = 0.28 for each tertile of uric acid). Longitudinal analysis of data from 186 women with follow-up examinations at a mean interval of 14.6 months also revealed no difference in reduction in both spine and femoral neck BMD between tertile groups of serum uric acid (the median BMD reduction for spine: -0.02, 0.01, -0.04, p = 0.95; the median BMD reduction for femoral neck: 0.008, 0.005, -0.003, p = 0.34). CONCLUSIONS: Serum uric acid level is not associated with BMD in postmenopausal women.

Ischemia-induced Netrin-4 promotes neovascularization through endothelial progenitor cell activation via Unc-5 Netrin receptor B.

Endothelial progenitor cells (EPCs) promote neovascularization and tissue repair by migrating to vascular injury sites; therefore, factors that enhance EPC homing to damaged tissues are of interest. Here, we provide evidence of the prominent role of the Netrin-4 (NTN4)-Unc-5 Netrin receptor B (UNC5B) axis in EPC-specific promotion of ischemic neovascularization. Our results showed that NTN4 promoted the proliferation, chemotactic migration, and paracrine effects of small EPCs (SEPCs) and significantly increased the incorporation of large EPCs (LEPCs) into tubule networks. Additionally, NTN4 prominently augmented neovascularization in mice with hindlimb ischemia by increasing the homing of exogenously transplanted EPCs to the ischemic limb and incorporating EPCs into vessels. Moreover, silencing of UNC5B, an NTN4 receptor, abrogated the NTN4-induced cellular activities of SEPCs in vitro and blood-flow recovery and neovascularization in vivo in ischemic muscle by reducing EPC homing and incorporation. These findings suggest NTN4 as an EPC-based therapy for treating angiogenesis-dependent diseases.

Two distinct cellular pathways leading to endothelial cell cytotoxicity by silica nanoparticle size.

BACKGROUND: Silica nanoparticles (SiNPs) are widely used for biosensing and diagnostics, and for the targeted delivery of therapeutic agents. Safety concerns about the biomedical and clinical applications of SiNPs have been raised, necessitating analysis of the effects of their intrinsic properties, such as sizes, shapes, and surface physicochemical characteristics, on human health to minimize risk in biomedical applications. In particular, SiNP size-associated toxicological effects, and the underlying molecular mechanisms in the vascular endothelium remain unclear. This study aimed to elucidate the detailed mechanisms underlying the cellular response to exposure to trace amounts of SiNPs and to determine applicable size criteria for biomedical application. METHODS: To clarify whether these SiNP-mediated cytotoxicity due to induction of apoptosis or necrosis, human ECs were treated with SiNPs of four different non-overlapping sizes under low serum-containing condition, stained with annexin V and propidium iodide (PI), and subjected to flow cytometric analysis (FACS). Two types of cell death mechanisms were assessed in terms of production of reactive oxygen species (ROS), endoplasmic reticulum (ER) stress induction, and autophagy activity. RESULTS: Spherical SiNPs had a diameter of 21.8 nm; this was further increased to 31.4, 42.9, and 56.7 nm. Hence, we investigated these effects in human endothelial cells (ECs) treated with these nanoparticles under overlap- or agglomerate-free conditions. The 20-nm SiNPs, but not SiNPs of other sizes, significantly induced apoptosis and necrosis. Surprisingly, the two types of cell death occurred independently and through different mechanisms. Apoptotic cell death resulted from ROS-mediated ER stress. Furthermore, autophagy-mediated necrotic cell death was induced through the PI3K/AKT/eNOS signaling axis. Together, the present results indicate that SiNPs within a diameter of < 20-nm pose greater risks to cells in terms of cytotoxic effects. CONCLUSION: These data provide novel insights into the size-dependence of the cytotoxic effects of silica nanoparticles and the underlying molecular mechanisms. The findings are expected to inform the applicable size range of SiNPs to ensure their safety in biomedical and clinical applications.

Müllerian inhibiting substance/anti-Müllerian hormone type II receptor protein and mRNA expression in the healthy and cancerous endometria.

Müllerian inhibiting substance/anti-Müllerian hormone (MIS/AMH) is a regulator of the female reproductive system, an indicator of ovarian reserve and a growth inhibitor of Müllerian duct-derived tumors in vivo and in vitro. The objective of the present study was to analyze MIS/AMH type II receptor (MIS/AMHRII) protein and mRNA expression in healthy human endometria compared with patients with endometrial hyperplasia and endometrial cancer, providing a foundation for MIS/AMH as a biological modifier for treatment of endometrial hyperplasia and endometrial cancer. The present study included healthy endometrial tissues (n=20), simple endometrial hyperplasia tissues without atypia (n=17), complex endometrial hyperplasia tissues without atypia (n=24) and endometrial cancer tissues (n=8). The location and variation of MIS/AMHRII protein expression was observed by immunohistochemistry. The expression was graded by two pathologists and was categorized as follows: Negative, weakly positive, moderately positive or strongly positive. Reverse transcription-quantitative polymerase chain reaction was used to quantify MIS/AMHRII mRNA expression. The expression of MIS/AMHRII protein was observed in the cytoplasm of healthy human endometria, endometrial hyperplasia and endometrial cancer cells. The frequency of MIS/AMHRII protein expression was 20.22±10.35% in the proliferative phase of the healthy endometrium and 24.09±11.73% in the secretory phase of the healthy endometrium. However, no differences were observed in the menstrual cycle phases. The frequency was 54.50±16.59% in endometrial hyperplasia without atypia, 55.10±15.87% in endometrial hyperplasia with atypia and 73.88±15.70% in endometrial cancer, indicating that expression was enhanced as the disease progressed from healthy to malignant status. In endometrial hyperplasia, MIS/AMHRII protein expression was significantly associated with histological complexity compared with atypia status. The present study demonstrated that MIS/AMHRII is present in healthy endometria, endometrial hyperplasia and endometrial cancer. The low expression frequency of MIS/AMHRII was not significantly different among normal endometrial tissues, however, the protein expression was elevated in endometrial hyperplasia and endometrial cancer. These findings indicated that the study of bioactive MIS/AMH, as a possible treatment for tumors expressing the MIS/AMH receptor, is essential.

Laparoendoscopic Single-Site Total Laparoscopic Hysterectomy: Clinical Factors that Affect Operative Times and Techniques to Overcome Difficulties.

STUDY OBJECTIVE: To compare surgical outcomes of patients who underwent laparoendoscopic single-site total laparoscopic hysterectomy (LESS-TLH) with operative times < 150 minutes and ≥150 minutes to determine the clinical factors that influence operative times. We also describe techniques that help overcome difficulties involved in this procedure. DESIGN: Retrospective study (Canadian Task Force classification II-2). SETTING: University medical center. PATIENTS: Two hundred thirty-four patients underwent LESS-TLH for benign uterine disease and cervical disease between September 2011 and February 2015. Thirty-seven patients (15.8%) were excluded from analysis. One hundred ninety-seven patients were divided into 2 groups according to the total operative time (median, 150 minutes): <150 minutes (n = 93) and ≥150 minutes (n = 104). INTERVENTIONS: LESS-TLH was performed using anterior, lateral, and posterolateral colpotomy techniques, and knife-in-bag morcellation was used for specimen extraction. MEASUREMENTS AND MAIN RESULTS: Patient characteristics, except for body mass index (BMI) and age, were generally similar in the 2 groups. Age, BMI, adhesiolysis, time to completion of colpotomy, stump repair time, specimen extraction time, blood loss, and weight of the uterus showed statistically significant differences between the 2 groups. Clinical factors that affected operative time were adhesiolysis of the posterior uterus (p = .010), time to completion of colpotomy (>65 minutes; p = .000), specimen extraction time (>34.4 minutes; p = .000), and weight of the uterus (>270 g; p = .015). Conversion to additional port laparoscopy occurred in 14 patients (5.98%). Conversion to laparotomy occurred in 1 patient (.43%). The surgical complication rates were 3.2% (3 patients) in the <150 minutes group and 3.8% (4 patients) in the ≥150 minutes group. No urologic injuries occurred in either group. One postoperative ileus occurred in the ≥150 minutes group; it was relieved by conservative treatment. CONCLUSION: Time to completion of colpotomy was influenced by adhesiolysis of the posterior uterus and specimen extraction time by weight of the uterus. Alternative methods for decreasing the time required to extract a large uterus and for approaching posterior adhesion of the uterus are needed for LESS-TLH.

Robot-assisted laparoscopic myomectomy for deep intramural myomas.

BACKGROUND: To evaluate the efficacy of robot-assisted laparoscopic myomectomy for deep intramural myomas. METHODS: We have conducted a retrospective study for 170 patients who underwent robot-assisted laparoscopic myomectomy by a single operator of tertiary university hospital. RESULTS: There were 100 cases of robot-assisted laparoscopic myomectomy for deep intramural myomas. The patients had 3.8±3.5 myomas on average, and the mean size of the largest myoma of each patient was 7.5±2.1 centimeters in diameter. Mean operative time was 276.4±97.1 minutes, and mean console time was 146.0±62.7 minutes. Thirty two patients had surgeries for other gynecologic conditions such as pelvic endometriosis or endometrial polyps along with myomectomy at the same time. All the patients recovered without any major complication. After the surgery, nine(75.0 %) of the 12 women pursuing a pregnancy became pregnant. CONCLUSION: Robot-assisted laparoscopic myomectomy for deep intramural myomas could be a minimal invasive surgical option for women who wish preserve fertility. Copyright © 2016 John Wiley & Sons, Ltd.

Graphene oxide induces apoptotic cell death in endothelial cells by activating autophagy via calcium-dependent phosphorylation of c-Jun N-terminal kinases.

Despite the rapid expansion of the biomedical applications of graphene oxide (GO), safety issues related to GO, particularly with regard to its effects on vascular endothelial cells (ECs), have been poorly evaluated. To explore possible GO-mediated vasculature cytotoxicity and determine lateral GO size relevance, we constructed four types of GO: micrometer-sized GO (MGO; 1089.9±135.3nm), submicrometer-sized GO (SGO; 390.2±51.4nm), nanometer-sized GO (NGO; 65.5±16.3nm), and graphene quantum dots (GQDs). All types but GQD showed a significant decrease in cellular viability in a dose-dependent manner. Notably, SGO or NGO, but not MGO, potently induced apoptosis while causing no detectable necrosis. Subsequently, SGO or NGO markedly induced autophagy through a process dependent on the c-Jun N-terminal kinase (JNK)-mediated phosphorylation of B-cell lymphoma 2 (Bcl-2), leading to the dissociation of Beclin-1 from the Beclin-1-Bcl-2 complex. Autophagy suppression attenuated the SGO- or NGO-induced apoptotic cell death of ECs, suggesting that SGO- or NGO-induced cytotoxicity is associated with autophagy. Moreover, SGO or NGO significantly induced increased intracellular calcium ion (Ca2+) levels. Intracellular Ca2+ chelation with BAPTA-AM significantly attenuated microtubule-associated protein 1A/1B-light chain 3-II accumulation and JNK phosphorylation, resulting in reduced autophagy. Furthermore, we found that SGO or NGO induced Ca2+ release from the endoplasmic reticulum through the PLC ß3/IP3/IP3R signaling axis. These results elucidate the mechanism underlying the size-dependent cytotoxicity of GOs in the vasculature and may facilitate the development of a safer biomedical application of GOs. STATEMENT OF SIGNIFICANCE: Graphene oxide (GO) have received considerable attention with respect to their utilization in biomedical applications. However, GO-related safety issues concerning human vasculature are very limited. In this manuscript, we report for the first time the differential size-related biological effects of GOs on endothelial cells (ECs). Notably, Subnanometer- and nanometersized GOs induce apoptotic death in ECs via autophagy activation. We propose a molecular mechanism for the GO-induced autophagic cell death through the PLCß3/IP3/Ca2+/JNK signaling axis. Our findings could be provide a better understanding of the GO sizedependent cytotoxicity in vasculature and facilitate the future development of safer biomedical applications of GOs.

Melissa Officinalis L. Extracts Protect Human Retinal Pigment Epithelial Cells against Oxidative Stress-Induced Apoptosis.

BACKGROUND: We evaluated the protective effect of ALS-L1023, an extract of Melissa officinalis L. (Labiatae; lemon balm) against oxidative stress-induced apoptosis in human retinal pigment epithelial cells (ARPE-19 cells). METHODS: ARPE-19 cells were incubated with ALS-L1023 for 24 h and then treated with hydrogen peroxide (H2O2). Oxidative stress-induced apoptosis and intracellular generation of reactive oxygen species (ROS) were assessed by flow cytometry. Caspase-3/7 activation and cleaved poly ADP-ribose polymerase (PARP) were measured to investigate the protective role of ALS-L1023 against apoptosis. The protective effect of ALS-L1023 against oxidative stress through activation of the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) was evaluated by Western blot analysis. RESULTS: ALS-L1023 clearly reduced H2O2-induced cell apoptosis and intracellular production of ROS. H2O2-induced oxidative stress increased caspase-3/7 activity and apoptotic PARP cleavage, which were significantly inhibited by ALS-L1023. Activation of the PI3K/Akt pathway was associated with the protective effect of ALS-L1023 on ARPE-19 cells. CONCLUSIONS: ALS-L1023 protected human RPE cells against oxidative damage. This suggests that ALS-L1023 has therapeutic potential for the prevention of dry age-related macular degeneration.

Caffeine Intake Is Associated with Urinary Incontinence in Korean Postmenopausal Women: Results from the Korean National Health and Nutrition Examination Survey.

INTRODUCTION: The objective of this study was to investigate whether caffeine intake is associated with urinary incontinence (UI) and quality of life (QOL) in Korean postmenopausal women. MATERIALS AND METHODS: We included 4,028 postmenopausal women who had participated in the Korea National Health and Nutrition Examination Survey IV (KNHANES IV). From the KNHANES questionnaire data, we ascertained the UI status of participants, defined as self-reported or medically diagnosed UI, and calculated their total daily caffeine intake through questions regarding the frequency of food consumption. The EuroQoL-5 Dimension (EQ-5D) descriptive system was used to evaluate QOL among the study population. RESULTS: The mean age of the study population was 63.19±0.25 years. Among the 4,028 women, the prevalence of medically diagnosed UI was 2.6% (n = 151), the prevalence of self-reported UI was 11.9% (n = 483), and the lifetime prevalence of UI was 15.8% (n = 639). In the study population, the presence of UI was not significantly different by age group, but daily caffeine consumption and the percentage of caffeine consumer decreased with age (P<0.001). Higher caffeine intake led to significantly higher prevalence of both medically diagnosed UI (p = 0.012) and self-reported UI (p = 0.040) in the study population. Even after adjusting for factors including age, parity, smoking status, hypertension and diabetes in logistic regression analysis, the positive association between caffeine intake and UI prevalence was observed in both medically diagnosed UI and self-reported UI (P = 0.017) among participants. In a subgroup analysis for EQ-5D (using continuous variables) in which we categorized participants into four groups according to UI presence and caffeine consumption, the EQ-5D scores were lower in the caffeine non-user group with UI than in the caffeine consumer group with or without UI. CONCLUSION: In a sample of Korean postmenopausal women, the prevalence of UI increased with higher caffeine consumption. Additionally, QOL was lower in caffeine non-users with UI than in the caffeine consumer groups. However, additional prospective studies are required to identify clear causation between caffeine consumption, UI prevalence and QOL.

A Comparison of Single-, Two- and Three-Port Laparoscopic Myomectomy.

BACKGROUND AND OBJECTIVE: A recent FDA safety communication has discouraged the use of a power morcellator for myoma extraction and has called for a change in surgical techniques for myomectomy. The objective of this study was to compare surgical outcomes of laparoscopic single-, two-, and conventional three-port myomectomy and to evaluate the feasibility of contained manual morcellation for uterine myoma. METHODS: This retrospective study was a review and analysis of data from 191 consecutive women who underwent single-, two-, or three-port myomectomy for the management of uterine myoma from January 1, 2009, through December 31, 2014. RESULTS: The 3 study groups did not differ demographically. Apart from operative time, the single- and two-port groups showed operative outcomes comparable to those of the multiport group. The single-port group had significantly longer operative times (P = .0053) than the two- and three-port groups. However, in the latter half of the single-port cases, the operative time was similar to those in the three-port group. The two-port surgery group showed a consistent operative time without a learning period. CONCLUSION: Single- or two-port myomectomy with transumbilical myoma morcellation is feasible and safe, with outcomes comparable to those of three-port myomectomy. These results suggest the potential for minimally invasive management of symptomatic uterine myoma, without the use of a power morcellator.

A comparison of postoperative pain after transumbilical single-port access and conventional three-port total laparoscopic hysterectomy: a randomized controlled trial.

INTRODUCTION: The objective of this study was to compare postoperative pain between single-port access total laparoscopic hysterectomy (SPA-TLH) using a transumbilical single-port system and conventional multi (three)-port access total laparoscopic hysterectomy (MPA-TLH). MATERIAL AND METHODS: A randomized controlled trial was conducted on 60 women who underwent SPA-TLH and MPA-TLH for benign gynecologic diseases between March 2014 and January 2015. Patients were randomly assigned to undergo SPA-TLH (n = 30) or MPA-TLH (n = 30). The variables measured included surgical outcomes and postoperative pain at 30 min and 1, 12, 24, and 48 h after surgery, assessed by the visual analog scale, bolus requirement of intravenous patient-controlled analgesia, and additional analgesic use. RESULTS: The two study groups did not differ in terms of patient demographics or surgical outcomes except for operative time. The SPA-TLH group had a longer operative time (p < 0.0001) compared with the MPA-TLH groups. There were no differences in pain scores between the two groups. The SPA-TLH group had significantly more intravenous analgesia requests during the 12-24 h after surgery (2.17 ± 3.05 vs. 0.79 ± 1.99; p = 0.047), more 24-48 h postoperative analgesics (0.21 ± 0.41 vs. 0.03 ± 0.19; p = 0.045), and more total additional analgesics (0.97 ± 0.94 vs. 0.45 ± 0.87; p = 0.034). CONCLUSION: SPA-TLH was feasible compared with MPA-TLH but the SPA-TLH group had a longer operative time. Although there is no difference in pain based on the visual analog scale pain score, the SPA-TLH group required more analgesia to give the same postoperative pain control.

Extension of the in vivo half-life of endostatin and its improved anti-tumor activities upon fusion to a humanized antibody against tumor-associated glycoprotein 72 in a mouse model of human colorectal carcinoma.

Endostatin is an endogenous angiogenesis inhibitor that exhibits potential anti-tumor efficacy in various preclinical animal models. However, its relatively short in vivo half-life and the long-term, frequent administration of high doses limit its widespread clinical use. In this study, we evaluated whether a fusion protein of murine endostatin (mEndo) to a humanized antibody against tumor-associated glycoprotein 72 (TAG-72), which is highly expressed in several human tumor tissues including colon cancer, can extend the serum half-life and improve the anti-tumor efficacy of endostatin by targeted delivery to the tumor mass. The fusion protein (3E8-mEndo) and mEndo showed improved anti-angiogenic activity in vitro and in vivo, predominantly by interfering with pro-angiogenic signaling triggered by vascular endothelial growth factor (VEGF). Moreover, in mice treated with 3E8-mEndo, we observed a markedly prolonged serum half-life and significantly inhibited tumor growth. The improved anti-tumor activity of 3E8-mEndo can be partially explained by increased local concentration in the tumor mass due to targeted delivery of 3E8-mEndo to implanted colon tumors. Collectively, our data clearly indicate that tumor-targeting antibody fusions to endostatin are a powerful strategy that improves the poor pharmacokinetic profile and anti-tumor efficacy of endostatin.

Stimulation of angiogenesis and survival of endothelial cells by human monoclonal Tie2 receptor antibody.

Angiopoietin-1 (Ang1) and its endothelium-specific receptor, tyrosine kinase with Ig and epidermal growth factor homology domain 2 (Tie2), play critical roles in vascular development. Although the Ang1/Tie2 system has been considered a promising target for therapeutic neovascularization, several imitations of large-scale production have hampered the development of recombinant Ang1 for therapeutics. In this study, we produced a fully human agonistic antibody against Tie2, designated 1-4h, and tested the applicability of 1-4h as an alternative to native Ang1 in therapeutic angiogenesis. 1-4h significantly enhanced the phosphorylation of Tie2 in a dose- and time-dependent manner in human Tie2-expressing HEK293 cells and human umbilical vein endothelial cells. Moreover, 1-4h induced the activation of Tie2-mediated intracellular signaling such as AKT, eNOS, MAPK, and Focal Adhesion Kinase p125(FAK). In addition, 1-4h increased the chemotactic motility and capillary-like tube formation of endothelial cells in vitro and enhanced the survival of serum-deprived endothelial cells. Taken together, our data clearly suggest that a human Tie2 agonistic antibody is a potentially useful therapeutic approach for the treatment of several ischemic diseases including delayed-wound healing and ischemic heart and limb diseases.

Usefulness of CA125 in the differential diagnosis of uterine adenomyosis and myoma.

OBJECTIVES: To evaluate the usefulness of CA125 levels in the differential diagnosis of adenomyosis and myoma. This has been addressed by few, if any, previous studies. STUDY DESIGN: Preoperative serum CA125 levels were measured in 2149 women who were diagnosed at total hysterectomy as having adenomyosis, myoma, endometriosis, and/or normal pelvis. Their medical records were retrieved and reviewed. RESULTS: The mean serum CA125 level in the adenomyosis patients was significantly greater than that in the patients diagnosed with myoma (65.21±96.60 U/mL vs.12.86±14.23 U/mL, respectively; P<0.001). In the differential diagnosis of adenomyosis and myoma, the cut-off serum CA125 level with the highest accuracy (78.8%) and highest diagnostic value (61.2%) was 19 U/mL. Using this cut-off value, the negative predictive value was 69.5%, and the positive predictive value was 76.5%. These results are clearly superior to those of the empirical single cut-off value of 35 U/mL. Receiver operating characteristic curve analysis revealed the area under the curve for differentiating adenomyosis from myoma was 0.776, indicating good diagnostic performance. CONCLUSION: In the differential diagnosis of adenomyosis and myoma, cut-off values for CA125, particularly the cut-off value of 19 U/mL, provide improved diagnostic performance. Serum CA125 testing can be performed during the initial screening of women with possible adenomyosis to differentiate this condition from myoma, although the diagnostic accuracy of using CA125 testing alone is limited.

Effect of helixor A on natural killer cell activity in endometriosis.

BACKGROUND AND AIM: NK cells are one of the major immune cells in endometriosis pathogenesis. While previous clinical studies have shown that helixor A to be an effective treatment for endometriosis, little is known about its mechanism of action, or its relationship with immune cells. The aim of this study is to investigate the effects of helixor A on Natural killer cell (NK cell) cytotoxicity in endometriosis MATERIALS AND METHODS: We performed an experimental study. Samples of peritoneal fluid were obtained from January 2011 to December 2011 from 50 women with endometriosis and 50 women with other benign ovarian cysts (control). Peritoneal fluid of normal control group and endometriosis group was collected during laparoscopy. Baseline cytotoxicity levels of NK cells were measured with the peritoneal fluid of control group and endometriosis group. Next, cytotoxicity of NK cells was evaluated before and after treatment with helixor A. NK-cell activity was determined based upon the expression of CD107a, as an activation marker. RESULTS: NK cells cytotoxicity was 79.38±2.13% in control cells, 75.55±2.89% in the control peritoneal fluid, 69.59±4.96% in endometriosis stage I/II endometriosis, and 63.88±5.75% in stage III/IV endometriosis. A significant difference in cytotoxicity was observed between the control cells and stage III/IV endometriosis, consistent with a significant decrease in the cytotoxicity of NK cells in advanced stages of endometriosis; these levels increased significantly after treatment with helixor A; 78.30% vs. 86.40% (p=0.003) in stage I/II endometriosis, and 73.67% vs. 84.54% (p=0.024) in stage III/IV. The percentage of cells expressing CD107a was increased significantly in each group after helixor A treatment; 0.59% vs. 1.10% (p=0.002) in stage I/II endometriosis, and 0.79% vs. 1.40% (p=0.014) in stage III/IV. CONCLUSIONS: Helixor A directly influenced NK-cell cytotoxicity through direct induction of CD107a expression. Our results open new role of helixor A as an imune modulation therapy, or in combination with hormonal agents, for the treatment of endometriosis.

Surgical resolution of obstructive dyspareunia after traumatic pelvic injury.

As more of the patients with traumatic pelvic injuries survive, they desire an optimal quality of life, including normal sexual function, even after the most severe injuries. We present the case of a 31-year-old woman who had dyspareunia due to impaired vaginal penetration after severe pelvic injury. After excision of a disunited fragment of pelvic bone and an adhesion band at the vaginal wall, dyspareunia was considerably resolved and the patient resumed sexual function. In cases of severe pelvic injury, physicians used to be satisfied with the patient's survival alone, and tended to regard sexual dysfunction as a trivial outcome. However, restoration of sexual function is an important part of management of these patients. In selected cases, obstructive dyspareunia resulting from traumatic pelvic injury can be managed by planned surgical intervention.