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Objectives: Oleanolic acid, a minor element of ginsenosides, and its derivatives have been shown to have cytotoxicity against some tumor cells. The impact of cytotoxic effect of oleanolic acid 3-acetate on ovarian cancer SKOV3 cells and endometrial cancer HEC-1A cells were examined both in vivo and in vitro to explore the underlying mechanisms. Methods: Cytotoxic effects of oleanolic acid 3-acetate were assessed by cell viability, phosphatidylserine exposure on the cell surface, mitochondrial release of cytochrome C, nuclear translocation of apoptosis-inducing factor, depolarization of mitochondrial transmembrane potential (ΔΨm), and generation of reactive oxygen species (ROS). In vivo inhibition of tumor growth was also assessed with xenografts in immunocompromised mice. Results: Oleanolic acid 3-acetate exhibited potent cytotoxicity toward SKOV3 and HEC-1A cells by decreasing cell viability in a concentration-dependent manner. Importantly, oleanolic acid 3-acetate effectively suppressed the growth of SKOV3 cell tumor xenografts in immunocompromised mice. Furthermore, oleanolic acid 3-acetate induced apoptotic cell death as revealed by loss of ΔΨm, release of cytochrome c, and nuclear translocation of apoptosis-inducing factor with a concomitant activation of many proapoptotic cellular components including poly(ADP-ribose) polymerase, Bcl-2, and caspases-8, caspase-3, and caspase-7. Oleanolic acid 3-acetate, however, caused a decrease in ROS production, suggesting the involvement of an ROS-independent pathway in oleanolic acid 3-acetate–induced apoptosis in SKOV3 and HEC-1A cells. Conclusion: These findings support the notion that oleanolic acid 3-acetate could be used as a potent anticancer supplementary agent against ovarian and endometrial cancer. Oleanolic acid 3-acetate exerts its proapoptotic effects through a rather unique molecular mechanism that involves an unconventional ROS-independent but mitochondria-mediated pathway. Keywords: Apoptosis, Endometrial cancer, Oleanolic acids, Ovarian cancer, Reactive oxygen species

To evaluate the viability of frozen embryos generated by intracytoplasmic sperm injection (ICSI) with frozen testicular spermatozoa.A total of 68 fresh embryo transfer (ET) cycles and 85 subsequent frozen-thawed ET (FET) cycles were grouped according to the source of spermatozoa: fresh testicular spermatozoa (TESE) or frozen-thawed testicular spermatozoa (t-TESE).There were no significant differences in the age of female patients, number of oocytes, or fertilization rates in fresh ET cycles with TESE (TESE-fresh ET) versus t-TESE (t-TESE-fresh ET). The rate of embryo survival after thawing (95.7 % vs. 94.0 %) was similar in frozen ET cycles (FET) with TESE (TESE-FET) and with t-TESE (t-TESE-FET). While there were significant differences in the proportion of good quality embryos, no statistical differences were found in the pregnancy or clinical abortion rates between the two groups. Moreover, delivery rates were not significantly different.Although the proportion of good quality embryos was affected by cryopreservation of testicular tissue, embryo survival rate was not. As well, subsequent pregnancy could be achieved successfully via t-TESE-FET cycles. Therefore, FET is not affected by the cryopreservation of testicular tissue, and avoids further oocyte retrieval and TESE procedures.

Citation Park D-W, Lee S-K, Hong SR, Han A-R, Kwak-Kim J, Yang KM. Expression of kisspeptin and its receptor GPR54 in the first trimester trophoblast of women with recurrent pregnancy loss (RPL). Am J Reprod Immunol 2012; 67: 132–139 Problem Kisspeptin and its receptor GPR54 play a major role in trophoblast invasion. The expression of kisspeptin and GPR54 in trophoblast and decidua and their relationship with decidual and peripheral blood natural killer (NK) cells are investigated in women with RPL. Method of study Trophoblast and decidual tissues were collected from 38 RPL women who miscarried a genetically normal fetus and 14 women who had elective abortion. Kisspeptin, GPR54, and decidual NK cells were investigated with immunohistochemistry, and peripheral blood NK cells were analyzed by flow cytometry. Results Kisspeptin expression in syncytiotrophoblast was significantly decreased in RPL women with normal (

Citation Park DW, Lee HJ, Park CW, Hong SR, Kwak-Kim J, Yang KM. Peripheral blood NK cells reflect changes in decidual NK cells in women with recurrent miscarriages. Am J Reprod Immunol 2010; 63: 173–180 Problem We aimed to investigate if peripheral blood natural killer (pNK) cell levels are correlated with decidual NK (dNK) cell levels, and if chemokine expression has any role in dNK cell regulation. Method of study Decidual tissues of women having two or more miscarriages with normal karyotype were collected after miscarriage and an immuno-histochemisty study was made. pNK cells were evaluated using flow cytometric analysis. Results The %CD3−/56+ and %CD3−/56+/16+ pNK cells showed a significant correlation with mean number of CD56+ dNK cells. The number of decidual CD16+ cells was significantly higher in women with elevated pNK (≥15%) than that of normal pNK (

Oxidative stress is an imbalance between free radicals and antioxidant molecules that can play an important role in the pathogenesis of iron‐deficiency anemia (IDA). The aim of this study was to investigate oxidative status in patients with IDA and alteration of oxidative status after iron treatment. Thirty‐three female patients with IDA and 25 healthy controls were included in this study. Oxidant and total antioxidant capacity were determined using free oxygen radicals test and free oxygen radicals defence (Form CR 3000, Callegari, Parma, Italy). Catalase activity was measured by spectrophotometer using a commercially available kit (Bioxytech Catalase‐520, OxisResearch, Portland, OR). Oxidant activity in patients with IDA was significantly higher than controls (P0.05). In conclusion, our data indicate that blood reactive oxygen species was lower and total antioxidant and catalase activity were higher after rather than before treatment in patients with IDA. The results of our study support the higher oxidative stress hypothesis in IDA; however, due to the limited number of cases included, more studies may be required to confirm the results. J. Clin. Lab. Anal. 23:319–323, 2009. © 2009 Wiley‐Liss, Inc.

Objective To investigate the effect of transforming growth factor (TGF)-β1 on the extracellular signal-regulated kinase (ERK) and Smad pathway and the role of peroxisome proliferator-activated receptor (PPAR)-γ in cultured human endometrial stromal cells. Design Experimental study. Setting Infertility center of a tertiary university hospital. Material(s) Human endometrial tissues obtained by hysterectomy from patients with conditions other than endometrial diseases. Intervention(s) Endometrial stromal cells were cultured under normal laboratory conditions. TGF-β1, rosiglitazone (PPARγ agonist), and PD98059 (ERK inhibitor) were added to endometrial stromal cell culture according to experimental purposes. Main Outcome Measure(s) Cell count, PRL expression, Smad and ERK phosphorylation, cyclooxygenase (COX)-2 expression, and prostaglandin E2 (PGE2) release. Result(s) TGF-β1 inhibited cellular proliferation and induced the expressions of COX-2, PGE2, and PRL of cultured human endometrial stromal cells. These effects may be mediated by Smad and ERK phosphorylation. Treatment with rosiglitazone, a PPARγ agonist, reversed the TGF-β1 effect by antagonizing the activation of ERK and Smad that was induced by TGF-β1. Conclusion(s) PPARγ plays a negative role by directly acting on Smad and ERK phosphorylation in human endometrial cell decidualization that is induced by TGF-β1 in vitro.

Objective To assess the effectiveness of the three-dimensional culture of spermatogenic cells in a collagen gel matrix from nonobstructive azoospermic patients and examine the relation between the success rate of in vitro spermatogenesis and serum FSH level as a diagnostic prediction. Design Prospective study using radioimmunoassay, immunocytochemistry, and flow cytometry with primary cultured cells. Setting Gynecologic clinics and human reproduction research laboratory. Patient(s) Primary culture of spermatogenic cells established from 18 nonobstructive azoospermic patients who underwent histologic diagnoses. Intervention(s) Primary culture of spermatogenic cells in a collagen-based gel matrix, subjected to immunological and flow cytometric analyses. Main Outcome Measure(s) In vitro culture of spermatogenic cells was established in an extracellular milieu that more closely resembled the in vivo condition. The number of chromosomes in newly generated cells during culture was determined by fluorescence-activated cell sorter (FACS) and immunocytochemical analysis. Effects of FSH on the differentiation of the spermatogenic cells were measured. Result(s) Results of histologic studies indicated that 8 of 18 patients showed the spermatocyte arrest. Immunocytochemical and FACS analysis indicated that after 12 days in culture, haploid cells comprised 11%–37% of the cultured cell population with a characteristic expression of a cellular marker for spermatids. The serum level of FSH appeared to be closely correlated with an increase in the number of haploid cells in culture. Conclusion(s) The present three-dimensional culture in a collagen gel matrix provides a suitable means by which spermatocytes could be induced to differentiate into presumptive spermatids in vitro. In addition, the plasma FSH level could be a good indicator for the success of differentiation of cultured spermatogenic cells obtained from patients with spermatogenic arrest.

A three-dimensional (3-D) endometrium culture was established, in which human endometrial stromal cells embedded in a mixture of collagen I, a major component of extracellular matrix, and matrigel, a basement membrane material, supports the epithelial cells seeded on top of the collagen/matrigel matrix. The biological growth and differentiation of the epithelial cells were studied microscopically and immunohistochemically. Transmission electron microscopy showed a polarized columnar epithelium in monolayer with basally positioned nuclei. Scanning electron microscopy revealed a confluent epithelium with an abundance of microvilli and cilia as well as pinopodes on the apical surface. An immunohistochemical staining showed that integrin alpha1, alpha4, and beta3 were co-localized with cytokeratin, confirming the epithelial origin of the cells. In contrast, immunoreactivity against cyclooxygenase-1 or -2 was positive in both epithelial and stromal cells. When epithelial cells were replaced by KLE cells, an endometrial cancer cell of epithelial origin, invasion of KLE cells into the stromal fraction was observed. The invasion was closely correlated to expression of matrix metalloproteinases and their tissue inhibitors of metalloproteinases in a manner consistent with paracrine fashion. The present 3-D culture imitates the normal endometrium physiologically as well as morphologically, thus provides an excellent in vitro tissue suitable for reproducing in vivo physiological processes, including endometrial cancer invasion.

Objectives. In this study, we have established an in vitro three- dimensional (3D) coculture, where normal endometrial stromal cells and endometrial cancer cells were cocultured under defined hormonal conditions, to investigate the potential paracrine effects on synthesis and secretion of matrix metalloproteinases (MMPs) and, thus, cancer invasion. Methods. Endometrial stromal cells were obtained by biopsy, cultured in the presence of 100 nM estrogen for 3 days, and then mixed with extracellular matrix (ECM) composed of collagen I and matrigel in a 4:1 ratio. After 3 more days in culture, a 3D coculture was established with HEC-1A cells, an endometrial adenocarcinoma cell line, grown on top of the mixture under various ovarian steroids (no steroid, 100 nM b-estradiol (E2), or 1.0 mM progesterone (P4)) for 10 days. The expression and local- ization of MMP-2, MMP-9, and integrin b3 subunit were visual- ized by immunocytochemistry and analyzed by reverse transcrip- tion polymerase chain reaction (RT-PCR). The extent of cancer invasion was quantified by Boyden's chamber assay. Results. Integrin b3 subunit was localized along the cell surface of HEC-1A cell under all three hormonal conditions. MMP-2 was located in the cytoplasm of stromal cells and on the surface of HEC-1A cells. Synthesis and secretion of stromal MMP-2 were increased in the presence of ovarian steroids. In contrast, expres- sion of stromal MMP-9 was suppressed in the presence of ovarian steroids. No MMPs were synthesized in HEC-1A cells. Invasion assay revealed that HEC-1A cells achieved high tumoral invasive- ness in the presence of b-estradiol. Conclusions. These findings suggest that stromal cell-derived MMP-2 is translocated to the surface of HEC-1A cells. Integrin b3 subunit might contribute, in part, to providing a binding site for MMP-2. Thus, HEC-1A cells invade by recruiting MMP-2 secreted by stromal cells, which is greatly enhanced in the presence of b-estradiol. The 3D coculture provides an excellent experimental system in which single parameters can be isolated from a complex in vivo system in the process of endometrial cancer invasion. © 2001 Academic Press

The task force of the Korean Society for Reproductive Immunology recommends intravenous immunoglobulin G treatment in women with reproductive failure, including recurrent pregnancy loss and/or repeated implantation failure, who show cellular immune factors such as abnormal natural killer cell levels, natural killer cell cytotoxicity, and/or type 1 T helper immunity.

Objective This study investigated the prevalence of infections with human papillomavirus, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalium in the semen of Korean infertile couples and their associations with sperm quality. Methods Semen specimens were collected from 400 men who underwent a fertility evaluation. Infection with above five pathogens was assessed in each specimen. Sperm quality was compared in the pathogen-infected group and the non-infected group. Results The infection rates of human papillomavirus, C. trachomatis, U. urealyticum, M. hominis, and M. genitalium in the study subjects were 1.57%, 0.79%, 16.80%, 4.46%, and 1.31%, respectively. The rate of morphological normality in the U. urealyticum-infected group was significantly lower than in those not infected with U. urealyticum. In a subgroup analysis of normozoospermic samples, the semen volume and the total sperm count in the pathogen-infected group were significantly lower than in the non-infected group. Conclusion Our results suggest that infection with U. urealyticum alone and any of the five sexually transmitted infections are likely to affect sperm morphology and semen volume, respectively.

Purpose Infection with Helicobacter pylori is an important risk factor for gastric cancer in humans. We compared the clinicopathologic features of gastric cancer patients based on H. pylori infection. Materials and Methods We prospectively studied 155 patients who had gastric cancer and underwent gastrectomies in 1 hospital in Korea. We examined H. pylori infections using the rapid urease test (RUT) with gastrectomy specimens and collected clinical and pathologic data. Results The number of H. pylori infections based on the RUT was 137 (88%). The H. pylori-negative group was significantly associated with AGC and tumor histology. H. pylori infection was significantly correlated with type I/IIa in EGC and type III/IV/V in AGC. AGC was significantly correlated with larger tumor size, lymphatic invasion, perineural invasion, and H. pylori infection based on univariate and multivariate analyses. Conclusions We report the prevalence of H. pylori based on the RUT in gastric cancer patients. H. pylori infection influences the tumor histology, progression, and growth type of gastric cancer.

Endometrial carcinoma is the most common neoplasm of the female genital tract, accounting for nearly one half of all gynecologic cancers in the Western world. Although intensive research on pathological phenomena of endometrial cancer is currently going on, but exact cause and biological aspects of this disease are not well described yet. In addition to well-documented roles of gonadotropin-releasing hormone (GnRH) in hypopituitary ovarian (HPO) axis, the agonistic or antagonistic analogs (or both) of GnRH have been shown to inhibit the proliferation of a variety of human gynecologic cancers. Thus, in the present study, we further examined the possibility that GnRH induces integrin beta3 and activation of focal adhesion kinase (FAK) through mitogen-activated protein kinases (MAPKs), ERK1/2 and p38, to inhibit the growth of HEC1A endometrial cancer cell line. As a result, both GnRH-I and GnRH-II resulted in a significant increase in integrin beta3 expression and evoked the activation of FAK in a time-dependent manner in these cells. In addition, these analogs induced an activation of ERK1/2 and p38 MAPK in a time-dependent manner as downstream pathways of FAK. It appears that GnRH-II has much greater effect on the activation of FAK, ERK1/2 and p38 compared to GnRH-I in these cells. Further, we demonstrated that the growth inhibition of HEC1A cells by GnRH-I or GnRH-II is involved in the activation of integrin-FAK and ERK1/2 and p38 MAPK pathways. Taken together, these results suggest that GnRH may be involved in the inhibition of endometrial cancer cell growth via activation of integrin beta3 and FAK as a direct effect. This knowledge could contribute to a better understanding of the mechanisms implicated in the therapeutic action of GnRH and its biomedical application for the treatment against endometrial cancer.

Objective: To investigate the role of extracellular adenosine triphosphatase (ATP) as an inducer of apoptotic cell death in human granulosa cells and to elucidate its underlying mechanism. Design: Prospective study. Setting: Gynecologic clinic and human reproduction research laboratory. Patient(s): Twenty-five patients undergoing IVF or IVF-ET. Intervention(s): ATP treatment of granulosa luteal cells subjected to primary culture. Main outcome measure(s): Apoptosis was assessed by the annexin V binding assay and the terminal deoxynucleotidyl transferase–mediated dUTP–digoxigenin nick-end labeling assay. The receptor type that binds ATP, thus mediating ATP-induced apoptosis, was determined by using the calcium imaging and patch-clamp techniques. Mitochondrial depolarization was assessed by staining with the membrane potential-sensitive dye 5,5′,6,6′-tetrachloro-1,1′,3,3′- tetraethylbenzimidazolyl carbocyanide iodide (JC-1). Result(s): ATP elevated [Ca 2+ ] i by mobilizing intracellularly stored Ca 2+ . An ionic membrane current evoked by ATP was mediated by the Ca 2+ -activated K + channel. ATP induced a mitochondrial depolarization with a concomitant increase in cellular apoptosis. Treatment with hCG eliminated both ATP-induced mitochondrial depolarization and apoptosis. Conclusion(s): Extracellular ATP recognized by P 2Y type purinoceptor triggers apoptosis in human granulosa luteal cells, and the downstream apoptotic cascade may act at least in part through mitochondria. The antiapoptotic effect of hCG has possible clinical implications in the treatment of disorders involving granulosa cell degeneration (such as follicular atresia).

Matrix metalloproteinase (MMP)-2 and -9 are secreted and translocated from endometrial stromal cells to HEC-1 A cells in a steroid-dependent manner. We investigated the paracrine effect of hepatocyte growth factor (HGF) on MMPs and metalloproteinase tissue inhibitor (TIMP) expression in stromal and endometrial cancer cells, and correlated with cancer cell invasiveness in three-dimensional (3D) coculture. The 3D coculture of endometrial stromal and cancer cell lines (HEC-1 A, HEC-IB, or KLE) were maintained in the presence or absence of HGF. The expression of MMP-2 and -9, MT1-MMP, TIMP-1 and -2 were examined by RT-PCR and zymography. Under the same conditions, invasion of the cancer cells was quantified by Boyden's chamber assay. HGF strongly induced MMP-9 mRNA expression in stromal cells, but had little effect on MMP-2 mRNA. MT1-MMP mRNA was detected only in KLE and stromal cells, which was also increased by HGF. TIMP-1 and -2 mRNAs was ubiquitous with no dependence on HGF. Zymographic analysis of MMPs showed that activation of MMP-2 and -9 was enhanced by HGF. A significant increase in invasion of all three cancer cells with HGF was observed. The effect of HGF on the invasiveness of 3D cocultured endometrial cancer cells and stromal cells appears to be due to induction of MMP-9 mRNA expression in stromal cells and /or increased activation of MMP-2 and MMP-9 by proteolytic digestion.

Matrix metalloproteinase (MMP)-2 and -9 are secreted and translocated from endometrial stromal cells to cancer cells in a steroid-dependent manner. We investigated the paracrine effect of hepatocyte growth factor (HGF) on the expression of MMPs and tissue inhibitor of metalloproteinases (TIMPs) in stromal and endometrial cancer cells, and correlated the results with cancer cell invasiveness in a three-dimensional (3D) coculture. The 3D coculture of endometrial stromal and cancer cell lines (HEC-1A, HEC-IB, or KLE) was maintained in the presence or absence of HGF. Invasion of the cancer cells was quantified by Boyden’s chamber assay. Under the same conditions, the expression of MMP-2 and -9, membrane type-1 matrix metalloproteinase (MT1-MMP), and TIMP-1 and -2 were examined by zymography and reverse transcription-polymerase chain reaction. A significant increase in the invasiveness of all three cancer cells in the presence of HGF was observed by Boyden’s chamber assay. HGF enhanced the activation of MMP-2 and -9 by zymography of MMPs. HGF strongly induced MMP-9 mRNA expression in stromal cells, but had little effect on MMP-2 mRNA. MT1-MMP mRNA was detected only in KLE and stromal cells, and was also increased by the presence of HGF. TIMP-1 and -2 mRNAs were ubiquitous, with no dependence on HGF.

Objective: The goal of this study was to evaluate the pregnancy potential of immature (metaphase I or germinal vesicle stage) oocytes retrieved in intracytoplasmic sperm injection (ICSI) cycles. Methods: A total of 1,871 couples with infertility underwent 2,984 ICSI cycles. Cycles in which three or fewer oocytes were retrieved were included in this study in order to evaluate the pregnancy potential of immature oocytes. Cycles were divided into five groups (group I–V), accord ing to the maturation status of the oocytes at the time of cumulus cell removal and ICSI. The fertilization and pregnancy rates after ICSI were analyzed and compared among the study groups based on the maturation status of the retrieved oocytes. Results: The retrieval of only immature oocytes was associated with a significant decrease in the fertilization rate (76.1% ± 37.3% vs. 49.0% ± 49.1%, 66.7% ± 48.7%; group I vs. group II, group III, respectively) and the average number of transferred embryos (1.5 ± 0.7 vs. 1.1 ± 0.4, 1.1 ± 0.6). The cycle cancellation rate was significantly higher when only immature oocytes were retrieved. The clinical pregnancy rate decreased significantly when the transferred embryos had originated from immature oocytes (16.9% vs. 10.3%, 1.2%). Conclusion: In ICSI cycles, the fertilization potential and pregnancy potential of the immature oocytes retrieved in ICSI cycles were inferior to those of mature oocytes. Therefore, increasing the number of injectable oocytes and transferrable embryos by using immature oocytes after their spontaneous in vitro maturation does not necessarily improve pregnancy outcomes.

Objective: Artificial oocyte activation (AOA) is an effective method to avoid total fertilization failure in human in vitro fertilization-embryo transfer (IVF-ET) cycles. AOA performed using a calcium ionophore can induce calcium oscillation in oocytes and initiate the fertilization process. We evaluated the usefulness of AOA with a calcium ionophore in cases of total fertilization failure in previous cycles and in cases of severe male factor infertility patients with non-motile spermatozoa after pentoxifylline (PF) treatment. Methods: The present study describes 29 intracytoplasmic sperm injection (ICSI)-AOA cycles involving male factor infertility at Cheil General Hospital from January 2006 to June 2013. Patients were divided into two groups (control, n = 480; AOA, n = 29) depending on whether or not AOA using a calcium ionophore (A23187) was performed after testicular sperm extraction-ICSI (TESE-ICSI). The AOA group was further split into subgroups according to sperm motility after PF treatment: i.e., motile sperm-injected (n = 12) and non-motile sperm-injected (n = 17) groups (total n = 29 cycles). Results: The good embryo rate (52.3% vs. 66.9%), pregnancy rate (20.7% vs. 52.1%), and delivery rate (10.3% vs. 40.8%) were lower in the PF/ AOA group than in the control group. When evaluating the effects of restoration of sperm motility after PF treatment on clinical outcomes there was no difference in fertilization rate (66.6% vs. 64.7% in non-motile and motile sperm, respectively), pregnancy rate (17.6% vs. 33.3%), or de livery rate (5.9% vs. 16.7%) between the two groups. Conclusion: We suggest that oocyte activation is a useful method to ensure fertilization in TESE-ICSI cycles regardless of restoration of sperm motility after PF treatment. AOA may be useful in selected patients who have a low fertilization rate or total fertilization failure.

Objective The presence of sperm-head vacuoles has been suspected to be deleterious to the outcomes of assisted reproductive technology (ART). It is difficult to accurately distinguish morphologically abnormal sperm with vacuoles under a light microscope. This study was performed to analyze the result of the observation of sperm-head vacuoles using Papanicolaou staining under a light microscope and whether the male partner's age affects these vacuoles. Methods Sperm morphology with vacuoles was evaluated using Papanicolaou staining and observed under a light microscope (400×) in 980 men. The normal morphology was divided into three categories (group A, 14% of normal morphology). The criteria for the sperm-head vacuoles were those given in the World Health Organization manual. For the analysis of the age factor, the participants were divided into the following groups: 26-30 years, 31-35 years, 36-40 years, 41-45 years, and 46-50 years. Results The percentage of sperm-head vacuoles increased with normal sperm morphology (group A vs. groups B, C) (p

Objective: The objective of this study is to estimate the effects of Inclear, a feminine cleanser, on sperm motility. Methods: Semen samples were obtained from infertile male patients. Following liquefaction, the raw semen samples were diluted with Ham’s F-10 nutrient mixture medium containing 0.4% human serum albumin solution at a ratio of 1:3. The semen samples were subsequently centrifuged to separate the seminal plasma from the serum. The supernatant was discarded, and the pellet was resuspended. The sample was again centrifuged to remove cell debris, and the supernatant was removed. The final pellet was gently loosened by resuspension and incubated in medium alone as a control, and in a 10% solution of the medium plus Inclear. A sampling time of 30 minutes was selected on the basis of sperm transport studies. Sperm motility was evaluated with computer-assisted sperm analysis. Results: A total of 20 samples were analyzed. The mean age of patients was 34.40 ± 2.96 years. There was no difference in sperm concentration and motility in the two samples at 0 minute and 30 minutes of incubation. In both semen samples, the sperm concentration and motility decreased after an incubation period of 30 minutes. However, there was no statistical difference between the samples. Sperm concentration and motility were not significantly different between the control and Inclear samples after 0 minute and 30 minutes of incubation. Conclusion: Inclear has no negative effects on sperm motility. This product can be recommended to pregnancy planners for vaginal hygiene and as a vaginal lubricant.

The ultimate function of the endometrium is to allow the implantation of a blastocyst and to support pregnancy. Cycles of tissue remodeling ensure that the endometrium is in a receptive state during the putative 'implantation window', the few days of each menstrual cycle when an appropriately developed blastocyst may be available to implant in the uterus. A successful pregnancy requires strict temporal regulation of maternal immune function to accommodate a semi-allogeneic embryo. To preparing immunological tolerance at the onset of implantation, tight temporal regulations are required between the immune and endocrine networks. This review will discuss about the action of steroid hormones on the human endometrium and particularly their role in regulating the inflammatory processes associated with endometrial receptivity.

Background and ObjectivesZZBiofilm is an encased colony of bacteria and the bacteria within the biofilm have resistance to antibiotics and the humoral immune system. Biofilm is cur rently thought to play an important role and to be very refractory to conventional therapeutics. The aim of this study is to analyze the relationship between the biofilm of the tonsil and recur rent tonsillitis with a case-control study. Subjects and MethodZZThis is a case-control study. The case group included 31 patients who had histories of tonsillitis and the control group included 29 volunteers who had no history of tonsillitis. Confocal laser scanning microscopy was performed to identify biofilms. We com pared the prevalence rates of biofilm between the 2 groups. Fluorescent in situ hybridization (FISH) was performed to identify bacteria, using probes for S. aureus, S. pyogenes, H. influ enzae, S. pneumoniae, and K. pneumonia. ResultsZZIn case group, biofilms were found in 22 (71.0%) of 31 and in control group, biofilms were found in 12 (41.4%) of 29. The prevalence rates of biofilm were significantly higher in case group (p=0.036). Twenty nine cases were enrolled in FISH and 12 cases of them were positive for FISH. S. aureus was most commonly identified. ConclusionZZThis study suggests that the biofilms could play a causative role in recurrent tonsillitis, further studies are needed to confirm pathogenic relation between biofilm and re current tonsillitis.�

We aimed to evaluate the effect of prednisolone (PDS) on natural killer cell (NK cell) cytolytic activity in vitro. Methods Blood samples from 74 patients with history of unexplained recurrent spontaneous abortion who elevated peripheral blood NK cell fraction were collected prospectively. Peripheral blood monocytes which containing NK cells were isolated and separated to three different tubes which containing target (K562) cells by the 50:1 effector to target (E:T) ratio. PDS or intravenous immunoglobulin (IVIG) was additionally added to 2 tubes for evaluate their suppressive effect. The percentage killing of target cells was recorded numerically by using flow cytometer and the values between groups were statistically analyzed. Results The mean target cell killing percentage was 40.5% in co-culture tube which was not added PDS or IVIG. In culture experiments which was added IVIG, the killing percentage is reduced to 37.7% which showed no significant differences compared to that of co-cultured tube which was not added PDS or IVIG. But, in experiments with added PDS, the killing percent was reduced to 19.5% and the difference was statistically significant ( P

Alterations of genes are known to be critical for the induction of tumorigenesis, but the mechanism of ovarian carcinogenesis is little understood and remains to be elucidated. In this study, we investigated the roles of brca1, brca2 and p53 genes in the development of ovarian cancer using conditional knockout mice generated by a Cre-loxP recombinant system. Following the application of recombinant adenovirus expressing Cre in vitro, the proliferation of ovarian surface epithelium (OSE) was increased. For instance, a significant increase in cell growth was observed in OSE cells in vitro by conditional knockout isolated from the mice bearing concurrent floxed copies of brca1 and brca2/p53. However, the proliferative effect of the ovarian cells was not observed in concurrent brca1/brca2 or p53 knockout mice in vivo, indicating that we could not observe the direct evidence of the involvement of brca1, brca2, and p53 in ovarian carcinogenesis. Since morphological changes including tumor formation were not observed in mice bearing floxed copies of concurrent brca1/brca2 or p53, the inactivation of brca1/2 or p53 is not sufficient for the induction of tumor formation. Taken together, these results suggest that the deficiency of these genes may not be involved directly in the mechanism of ovarian carcinogenesis.