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High-risk human papillomavirus (HPV) is associated with the carcinogenesis of not only cervical cancer but anal, penile, vulvar, vaginal, and oropharyngeal cancers. Although molecular biological mechanisms of high-risk HPV (HR-HPV)-associated carcinogenesis is well studied, it remains unclarified why cervical cancer is the most common among these HPV-associated cancers. Two major causes are that the cervix is a susceptible site to viral infection because of its immune deficiency to protect allogenic sperm in reproductive function and that the squamocolumnar junction (SCJ) where cervical neoplastic diseases develop is composed of tissue stem cells with self-renewal and pluripotency. This specific environment of the cervix allows HPVs to be persistently infected into the cervical epithelial cells, followed by the immortalization of the HPV-infected cells. We here focused on the carcinogenesis specific to the cervix as novel therapeutic strategies for cervical cancer, targeting cancer stem cells and mucosal immunotherapy.

Background: X-ray pelvimetry is used for evaluation of pelvic inlet generally to diagnose cephalopelvic disproportion (CPD) or contracted inlet. Cesarean section delivery (C/S) is often performed for labor dystocia without CPD or contracted inlet. Here, we examined whether X-ray pelvimetry is useful to decide on mode of delivery in women with dystocia.Method: A total of 1118 pregnant women received X-ray pelvimetry before or during labor. Overall, 205 women with cesarean deliveries for indications except for dystocia were excluded. The remaining 913 women undergoing induction/augmentation were retrospectively investigated. Obstetrical and maternal variables were analyzed by univariate, multivariate or ROC analysis.Results: Among 913 women, 37 (4%), including three with contracted inlet and seven with CPD, gave birth by C/S, whereas 876 gave birth by vaginal delivery. Low maternal height, older age, small obstetrical conjugate, large weight and infant head size were significantly associated with risk of C/S for dystocia. Multivariate analysis revealed that the obstetrical conjugate was an independent variable for risk of C/S. The area under the ROC curve (AUC) and the optimal cut-off values, respectively, were as follows: obstetrical conjugate: 0.68 and 11.7 cm (odds ratio=4.27), transverse diameter: 0.59 and 11.4 cm (odds ratio=1.82), maternal height: 0.70 and 155.5 cm (odds ratio=4.33), and maternal weight before pregnancy: 0.55 and 49.7 kg (odds ratio=1.98).Conclusion: The obstetrical conjugate was an independent variable associated with risk of C/S for dystocia. Additionally, maternal height was comparable to the conjugate in term of diagnostic ability. Our data suggested that routine X-ray pelvimetry was not beneficial to identify women at risk of C/S for dystocia.

Aim Although the procedure of abdominal trachelectomy has been remarkably improved, preventing subsequent cervical stenosis remains challenging. In this study, we analyzed the clinicopathological risk factors for cervical stenosis to explore the appropriate surgical procedures for the prevention of cervical stenosis following trachelectomy. Methods Thirty-two patients who underwent abdominal extended and radical trachelectomy were assessed retrospectively (median follow-up period = 33 months). To evaluate the risk factors, the clinicopathological factors were analyzed by univariate and multivariate analyses. The reconstructed uterine length (UtL), that is, the length between the vaginal end of the neo-cervix and the uterine fundus, was measured by transvaginal ultrasound after surgery. The cut-off value for the UtL was assessed by a receiver operating characteristic (ROC) curve analysis. Results Cervical stenosis of any grade was observed in 12 patients (grade 1 = 9, grade 3b = 3). Among the various clinicopathological factors, the UtL and cervical length (CL) were significantly related to cervical stenosis following trachelectomy. The multivariate analysis revealed that the UtL, but not CL, is an independent risk factor for stenosis. The ROC curve analysis revealed that stenosis was significantly more likely to occur in patients with a UtL shorter than 53 mm (area under the ROC curve = 0.902). UtL in the patients who became pregnant was longer than that in the patients who did not. No evidence of recurrent cancer was observed during the follow-up period. Conclusion Our proposed method may provide a functional reconstructed uterus with preserving fertility by remaining UtL more than 53 mm.

Aim. To investigate the changes in the maternal immune system at term pregnancy, we studied the expression of natural cytotoxicity receptors (NCRs) and the cytokine production of NK cells in term placenta decidua and peripheral blood. Methods. Term decidua and peripheral blood were taken from patients undergoing elective cesarean section. The lymphocytes were separated using density gradient centrifugation (DGC) from peripheral blood and were separated from decidua using DGC after enzyme digestion. These cells were stained with FITC anti-CD56 and Per-CP anti-CD3 monoclonal antibodies, and the NCRs were stained with PE-conjugated anti-NKG2D, NKp46, NKp30, and NKp44 monoclonal antibodies. Cytokines, including IFN-γ, TNF-α, IL-10, and TGF-β, were stained and then analyzed by flow cytometry. Results. There were fewer cells positive for NKG2D, NKp46, and NKp30 among CD56+CD3- cells in deciduas than in peripheral blood, but the percentages of NKp44-positive cells in CD56+CD3- lymphocytes in deciduas tended to be higher. Conclusion. The decreased expression of some NCRs in deciduas may be related to decreased cytotoxicity at term pregnancy, but the increased expression of NKp44 may affect the increased cytokine production in the decidua. Similarly, the expression of NCRs in the decidua may be connected to the maintenance of pregnancy at term.

Implantation is essential for successful reproduction. In humans, uterine receptivity is restricted to the mid-secretory phase, days 19 to 24 of the menstrual cycle; and this period is called the window of implantation (WOI). Large populations of decidual leukocytes infiltrate the implantation site, and the levels of cytokines, prostaglandins (PGs), and leukocytes are up-regulated in the endometrium during implantation. Implantation is like a local wound healing process which is characterized by a strong Th1, pro-inflammatory response in which high levels of pro-inflammatory cytokines such as IL-6, LIF, IL-8, and TNFα are involved. In addition, cyclooxygenase (COX)-2 expression increases between days 2 to 5, suggesting that PGs are necessary for the process of stromal cell decidualization in early pregnancy. Recently, it was reported that endometrial biopsies taken during spontaneous cycles that preceding IVF treatment improved the rate of implantation, clinical pregnancies, and live births in p...

Background/Aim: The aim of this study was to evaluate the gene and protein expression of angiotensin type (AT) 1, AT2 receptors in endometriotic lesions and its relation to prostaglandin (PG) synthases. Materials and Methods: Endometriosis samples were obtained from 32 patients with endometriotic cysts. Endometrial tissues were obtained during operations for benign gynecological conditions. The expression of the AT1 and AT2 receptor mRNA and that of PG-endoperoxide synthase 2 and microsomal PGE2 synthase-1 (mPGES-1) was examined by quantitative RT-PCR. Immunohistochemical staining was performed for these receptors. Results: AT1 and AT2 receptor proteins were mostly located in endometrial glandular epithelium and some stromal cells. Immunoreactivity of the receptor proteins was observed in both the eutopic endometrium and endometriotic lesions. The AT1/AT2 ratio in endometriotic cysts (median 7.29, range 1.88-187.60) was significantly increased compared with that in the eutopic endometrium in the proliferative-phase in controls (median 1.01, range 0.37-2.09, p < 0.001). There was a relationship between the AT1 mRNA expression and that of mPGES-1 mRNA in the endometriotic cysts (r = 0.394089, p < 0.05). There was a significant relationship between the mRNA expression of the AT2 receptor and that of mPGES-1 in eutopic endometrium of non-endometriotic control (r = 0.610714, p < 0.05). Conclusion: Renin-angiotensin system may play an important role in the pathophysiology of endometriosis.

Aim: Cytokines such as tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) play an important role in maintaining pregnancy. Toll-like receptors (TLRs) have also been associated with innate immune responses during pregnancy. Placenta growth factor (PlGF) is one of the angiogenic factors and soluble vascular endothelial growth factor receptor 1 (sVEGFR1) is one of the antiangiogenic factors that regulate PlGF function. To elucidate the effects of cytokines and TLR ligands on the production of angiogenic and antiangiogenic factors in trophoblasts, we examined the production of PlGF and sVEGFR1 from trophoblasts following stimulation with cytokines and TLR ligands. Methods: Villous tissues were obtained from healthy pregnant women who had had an artificial abortion. The trophoblasts were isolated from villous tissues. Subsequently, trophoblasts were treated with TNF-α, IFN-γ and TLR ligands. The levels of PlGF and sVEGFR1 were measured by enzyme-linked immunosorbent assay. The expression of those mRNA was analyzed using quantitative reverse transcription PCR. Results: The production of PlGF in trophoblasts increased by the addition of TNF-α or IFN-γ and decreased by TLR4 ligand. sVEGFR1 levels also increased by following the stimulation with IFN-γ or TLR ligand. Conclusions: Cytokines such as TNF-α and IFN-γ and TLR ligands may contribute to the production of angiogenic and antiangiogenic factors and may affect the placental formation.

Aim The anti-β2-GPI antibody (aβ2-GPIAb) has been detected in recurrent fetal loss with strong pathogenic activity. The effects of aβ2-GPIAb on cytokine production and aβ2-GPIAb binding sites in first-trimester trophoblast cells were evaluated. Methods First-trimester trophoblast cells were cultured in 24-well tissue culture plates with immunoglobulin G (IgG) obtained from aβ2-GPIAb-positive and aβ2-GPIAb-negative serum. Cytokines in the cultured supernatant were measured using the suspension array system and enzyme-linked immunosorbent assays. To identify potential binding sites for aβ2-GPIAb, such as toll-like receptors (TLR) 2 or TLR4, we used mouse monoclonal anti-TLR2 and/or anti-TLR4 antibodies to inhibit TLR and then measured cytokine production. Results The production of cytokines, such as interleukin-6 and interleukin-8, increased more in response to aβ2-GPIAb-positive IgG than to aβ2-GPIAb-negative IgG in trophoblast cells. The secretion of cytokines from trophoblast cells decreased when the TLR were blocked with mouse monoclonal anti-TLR2 and anti-TLR4 antibodies. Conclusion We suspect that aβ2-GPIAb might increase cytokine production by binding to TLR2 or TLR4. The increased cytokine production in response to aβ2-GPIAb might play a role in the increased inflammatory response in the placenta.

Preservation of fertility has been recommended for cancer-bearing patients of reproductive age before undergoing cancer treatment. However, there are many considerations and it is difficult to preserve fertility for all patients undergoing therapy for malignancies. Female cancer survivors had lower pregnancy and live birth rates compared with others that underwent assisted reproductive technologies (ARTs). We should continue to consider the issue of infertility in patients who underwent therapies for malignancies. This is the first report of a successful live birth in a patient with a cranial tumor who underwent radiotherapy and chemotherapy after implantation of an autologous embryo. The patient was a 27-year-old Japanese woman. She was diagnosed with suprasellar germinoma at 13 years of age, and she developed panhypopituitarism after radiotherapy and chemotherapy. At 27 years of age, she began infertility treatment with in-vitro fertilization (IVF). The level of anti-Mallerian hormone (AMH) was 4.29 ng/ml. After ovarian stimulation by high purified human menopausal gonadotropin (HP-hMG), she obtained two blastocysts and became pregnant by implantation of a cryopre- served blastocyst. At 37 gestational weeks, she delivered a healthy female baby by cesarean section.

To investigate the effects of the anti- β2-glycoprotein I (anti-β2-GPI) antibody on the tro- phoblast by evaluating the effects of the anti-β2-GPI antibody on the expression of toll-like receptor (TLR) mRNA in choriocarcinoma cells and primary trophoblast cells.We cul- tured the choriocarcinoma cells (BeWo) and the pri- mary first trimester tropho- blast with the IgGs taken from anti-β2-GPI antibody-positive and -negative sera. Four hours later we purified the RNAs from those cells. We measured the expressions of TLR mRNA in cells using real-time. PCR.The expression of TLR mRNA increased in BeWo cells and primary trophoblast cells cultured with the IgGs taken from anti-β2-GPI antibody-positive women. Specifically, the expression of TLR1, 2, and 4 in BeWo cells -and TLRI, 3, 4, and 9 in first trimester trophoblast cells increased significantly.The anti-β2-GPI antibody-positive IgG increased the TElR mRNA expression in choriocarcino- ma cells and primary trophoblast cells. We suggest that anti-β2-GPI antibodies may bind to trophoblast and increase the expression of TLR mRNA.

Background/Aim: We demonstrated that AT1 and AT2 are expressed and both pathways balance the renin-angiotensin system in endometriosis. MAS1, a specific receptor of angiotensin (1–7), opposes AT1 pathway-associated tissue remodelling. It is not known whether MAS1 has an effect on the pathogenesis of endometriosis or not. Materials and Methods: Ovarian endometriotic tissues (endo-Ov) and eutopic endometrial tissues (endo-Em) were obtained from 29 patients with endometrial cysts. Normal endometrial tissues (cont-Em) were obtained from patients without endometriosis. Immunohistochemical staining was performed for MAS1, AT1 and AT2 in the endometriosis-associated tissues. The mRNA levels of these receptors were examined by quantitative reverse transcription PCR. Results: MAS1 was immune-positive at the apical side of the glandular epithelium in the endometriotic lesions. The MAS1 mRNA levels in endo-Ov were increased significantly, irrespective of the menstrual cycle phase. The MAS1 mRNA levels were significantly higher in the proliferative-tissues of the endometriosis patients than in those of the controls. The ratio of the MAS1 to the AT1 mRNA in the proliferative tissues was increased predominantly in the endometriosis patients compared with that in the controls. Conclusion: High MAS1 expression in the endometrium might promote the initiation of endometriosis via migration of proliferative tissue.

The anti-β2-GPI antibody (aβ2-GPIAb) has been detected in recurrent fetal loss with strong pathogenic activity. The effects of aβ2-GPIAb on cytokine production and aβ2-GPIAb binding sites in first-trimester trophoblast cells were evaluated.First-trimester trophoblast cells were cultured in 24-well tissue culture plates with immunoglobulin G (IgG) obtained from aβ2-GPIAb-positive and aβ2-GPIAb-negative serum. Cytokines in the cultured supernatant were measured using the suspension array system and enzyme-linked immunosorbent assays. To identify potential binding sites for aβ2-GPIAb, such as toll-like receptors (TLR) 2 or TLR4, we used mouse monoclonal anti-TLR2 and/or anti-TLR4 antibodies to inhibit TLR and then measured cytokine production.The production of cytokines, such as interleukin-6 and interleukin-8, increased more in response to aβ2-GPIAb-positive IgG than to aβ2-GPIAb-negative IgG in trophoblast cells. The secretion of cytokines from trophoblast cells decreased when the TLR were blocked with mouse monoclonal anti-TLR2 and anti-TLR4 antibodies.We suspect that aβ2-GPIAb might increase cytokine production by binding to TLR2 or TLR4. The increased cytokine production in response to aβ2-GPIAb might play a role in the increased inflammatory response in the placenta.

Introduction Placenta growth factor (PlGF) is a growth factor originated from placenta. The sFlt-1 is soluble receptor for PlGF and suppresses PlGF function. It has been reported that in preeclampsia, serum level of PlGF decreased and sFlt-1 level increased and that preeclampsia placenta is in hypoxic condition. Metal-responsive transcription factor (MTF)-1, Hemoxigenase 1 (HO-1) and Hypoxia responsive factor -1 (HIF-1) may be induced in hypoxic condition. Objectives In order to investigate pathophysiology in preeclampsia, we studied the expression of PlGF, sFlt-1, MTF-1, HO-1 and HIF-1 alpha mRNAs in placenta taken from preeclampsia and the effect of preeclampsia sera on their expression of choriocarcinoma cells and analysed the effect of placental hypoxia and serum factor on the expression of PlGF and sFlt-1 mRNA. Methods Placenta and serum samples were taken from preeclampsia and normal pregnancy with informed consent. The choriocarcinoma cells (JEG-3) were cultured in 24-well tissue culture plate. The cells were cultured with preeclampsia and normal pregnant sera. The RNAs were purified from these cells 24h after and placenta. The expressions of these mRNA were measured by using the real time PCR method (Applied Biosystems-7500). Results The expression of PlGF mRNA decreased and that of sFlt-1mRNA increased in preeclampsia placenta. The expression of MTF-1 and HO-1 mRNA decreased. The correlation was found between the expression of PlGF and MTF-1 mRNA, PlGF and HO-1 mRNA and sFlt-1 and HO-1mRNA. Moreover, expression of sFlt-1mRNA increased and HO-1mRNA decreased in JEG-3 cells after incubation of preeclampsia sera. Conclusion The changes of PlGFmRNA in preeclampsia placenta may relate to the expression of MTF-1 and HO-1 mRNA. The changes of sFlt-1mRNA may relate to the expression of HO-1 mRNA and serum factor. Not only hypoxia but also serum factor may play a role of the levels of PlGF and sFlt-1 in preeclampsia placenta.

Objectives The soluble endoglin (sEng) is an antiangiogenic protein inhibiting TGF- β signaling and eNOS activation. It is well known the levels of sEng increase in preeclamptic patients. However, the factors increasing the sEng in preeclampsia remains poorly understood. In preeclampsia, many autoantibodies such as antiphospholipid, angiotensin II type IA receptor autoantibody (AT1-AA) have been reported. To investigate factors increasing sEng in preeclampsia, we studied the effect of IgG from preeclampsia sera on the production of sEng from human trophoblast cells. Methods Serum samples were from women with normal pregnancies and those with preeclampsia. Villous tissues were from healthy pregnant women having artificial abortion. IgGs isolated from normal pregnancy and preeclampsia sera were eluted by Protein G affinity chromatography. To investigate the possibility of AT1-AA, the production and mRNA expression of sEng were evaluated by AT1-receptor antagonist (losaltan). We also analyzed the various kinds of cytokine productions with or without losartan to find the relationship between sEng, AT1-AA and cytokine productions. Results The addition of preeclampsia IgG into primary trophoblast led to increased release of sEng and increased expression of Eng mRNA. The increased production and mRNA expression of sEng were blocked by the treatment of losartan. The production of TGF- β was increased under the presence of losartan. We considered this effect comes from the decreased production of sEng with losaltan. Conclusions We suggest that AT1-AA in IgG fraction from preeclampsia may play an important role of high level of serum sEng in preeclampsia. Disclosures Y. Koabayshi: None. T. Yamamoto: None. F. Chishima: None. T. Murase: None. H. Azuma: None. A. Nakamura: None. H. Takahashi: None.

48h. Immortalizedfirst trimester human trophoblastHTR8 cells were plated on matrigel chambers with various concentrations ofP. gingivalis sup.NK-92MI cellswere cultured withP. gingivalis supandcytotoxicityagainstK562cells and TNFproductionwere estimatedwithflowcytometry and ELISA respectively. Results: Matrigel invasion of HTR8 was inhibited by P. gingivalis sup in a concentration dependent manner. The inhibition was observed at non-cytotoxic concentrations. P. gingivalis sup also induced de novo production of TNFbymucosal CD56+ NK cells but was not sufficient to induce natural killer activities. Conclusions: Culture supernatant of P. gingivalis contains inhibitory substance(s) on early trophoblast invasion into myometrium and inflammatory cytokine production of mucosal natural killer cells. Our observations suggest very early events in human pregnancy are susceptible for periodontitis through direct functional suppression of trophoblastic invasion and inflammatory cytokine production by NK cells. In this sense, dental care for periodontitis must be done before pregnancy to reduce pregnancy complications.