Trans-vaginal specimen extraction following totally laparoscopic subtotal gastrectomy in early gastric cancer.

BACKGROUND: Although natural orifice extraction is now widely performed, there have been no reports of this procedure following subtotal gastrectomy for gastric cancer. This report describes trans-vaginal specimen extraction in four patients with early gastric cancer. METHODS: The clinical data of four patients with early gastric cancer were reviewed. Totally laparoscopic subtotal gastrectomy and D1 + ß lymph node dissection was performed using five trocars and a conventional procedure. Posterior colpotomy was performed by an experienced gynecologist, who retrieved the specimens in a retrieval bag via the trans-vaginal route. The colpotomy site was repaired immediately following specimen removal. Reconstruction was performed using the intracorporeal Billroth II method and an endo-GIA 60. RESULTS: Totally laparoscopic subtotal gastrectomy and trans-vaginal specimen extraction was successfully accomplished in all patients without intraoperative complications. CONCLUSIONS: The present technique may be a safe and feasible operative procedure for some limited groups of elderly female patients with early gastric cancer.

Proteomic analysis reveals an elevated expression of heat shock protein 27 in preeclamptic placentas.

AIM: The purpose of this study was to identify the placental proteins that are associated with preeclampsia by performing proteomic analysis. METHODS: To identify the proteins associated with preeclampsia, we performed two-dimensional electrophoresis (2-DE), followed by silver staining. The overexpressed proteins were identified by performing matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), followed by peptide mass fingerprinting, a protein database search and Western blot analysis. Immunohistochemical staining was performed to determine the localization of the overexpressed Hsp27. RESULTS: By use of 2-DE and MALDI-TOF-MS analysis, twelve differentially expressed proteins were identified, of which four proteins were upregulated and eight proteins were downregulated. One of the upregulated spots was identified as Hsp27. Immunohistochemical analysis showed that Hsp27 was mainly located in the trophoblasts. The Western blot analysis showed that the expression of Hsp27 in the tissues of the preeclampsia placenta was significantly increased. CONCLUSIONS: Our study confirmed that four proteins are upregulated and eight proteins are downregulated in preeclampsia. These differentially expressed proteins include signal transduction protein and molecular chaperon protein, in which Hsp27 is upregulated. We suggest that the increased expression level of Hsp27 might be correlated with the pathophysiology of preeclampsia.

Cushing's syndrome in pregnancy with a severe maternal complication: a case report.

Cushing's syndrome (CS) in pregnancy may be confused with a complication of pregnancy, such as pre-eclampsia or gestational diabetes. We managed a case of CS in pregnancy that was considered to be severe pre-eclampsia due to uncontrolled hypertension. The fetus was delivered via emergency cesarean section at 31 weeks' gestation because of severe pre-eclampsia and pulmonary edema. The parturient was admitted to the intensive care unit for severe maternal complications, including pulmonary hemorrhage, acute renal failure, disseminated intravascular coagulopathy, and congestive heart failure. A spine magnetic resonance image and 99m-technetium whole-body scan obtained postpartum showed multiple thoracolumbar spine compression fractures (Deleted; t-2,5,8,10,11, and -12; and L-1,2,3,4, and -5), multiple rib fractures, and a left iliac bone fracture due to osteoporosis. As a result of diagnosing CS after delivery, an adrenal cortical adenoma of the right adrenal gland was demonstrated and a laparoscopic adrenalectomy was successfully performed.

Failure of uterine artery embolization for controlling postpartum hemorrhage.

AIM: We evaluated the efficacy of uterine artery embolization (UAE) for controlling postpartum hemorrhage (PPH). MATERIALS AND METHODS: Between January 2008 and December 2009, 23 women with intractable PPH underwent UAE. Specific diagnoses included uterine atony (n = 10), placenta accreta (n = 8), puerperal hematoma (n = 2) and placental polyp (n = 3). RESULTS: Of 10 patients with uterine atony, treatment with UAE failed in two women with severe vasoconstriction. One patient developed lumbosacral plexopathy. All eight patients with placenta accreta were treated successfully with the placement of multiple sutures in the placental bed and UAE. Two of the three women with placental polyps were treated successfully with UAE and packing of the uterus. CONCLUSIONS: Embolization should follow resuscitation for vascular collapse. In the case of an adherent placenta, embolization is more effective with the placement of multiple sutures in the placental bed or compression of the placental bed.

Increased interaction between heat shock protein 27 and mitogen-activated protein kinase (p38 and extracellular signal-regulated kinase) in pre-eclamptic placentas.

AIMS: Heat shock protein 27 (Hsp27) is a well-known stress response protein that is characterized by its phosphorylative capacity. Hsp27 becomes phosphorylated in response to various stimuli through interaction with several different kinases. The purpose of this study was to evaluate the interaction between Hsp27 and mitogen-activated protein kinase (MAPK) (p38, extracellular signal-regulated kinase [ERK], and c-Jun N-terminal kinase) in the human placenta derived from patients with pre-eclampsia. METHODS: Western blot analysis was used to examine the levels of expression of Hsp27 and MAPK (p38, ERK, and c-Jun N-terminal kinase). Immunoprecipitation analysis was used to determine the interaction between Hsp27 and MAPK (p38 and ERK). RESULTS: Western blotting analysis and immunohistochemistry showed that the expression of Hsp27 and p-Hsp27 in the placental tissues of the pre-eclampsia group were significantly higher than that in the normal pregnancy group. Immunoprecipitation analysis showed that the interaction between Hsp27 and MAPK (p38 and ERK) was significantly increased in the pre-eclamptic placenta tissues. CONCLUSION: The interaction between Hsp27 and MAPK was increased, suggesting that phosphorylation of Hsp27 might be induced by p38 and ERK in placentas from patients with pre-eclampsia.

Altered gene expression of caspase-10, death receptor-3 and IGFBP-3 in preeclamptic placentas.

Enhanced apoptosis has been observed in the placentas of women with preeclampsia, but few studies have examined changes at the molecular level. This study was designed to detect genes specifically expressed in full-term preeclamptic placentas. Tissue samples were collected immediately after cesarean delivery from 11 normal and 8 preeclamptic placentas at 35-40 weeks of gestation. Total RNAs were extracted and hybridized to a cDNA microarray. Results were confirmed by reverse-transcription polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry. Hematoxylin and eosin and TUNEL staining were also performed to confirm apoptosis in preeclamptic placentas. Among 205 genes, three were up- or down-regulated in preeclamptic placentas. The expression of caspase-10 and death receptor 3 (DR-3) was significantly increased, whereas insulin-like growth factor binding protein-3 (IGFBP-3) was strongly down-regulated. RT-PCR analysis and Western blotting confirmed these effects. Immunohistochemical analysis showed that the DR-3, caspase-10 and IGFBP-3 proteins were localized in the syncytial membrane. Apoptosis in the trophoblast was also increased in term placentas from women with pregnancies complicated by preeclampsia. These results suggest that caspase-10, DR-3 and IGFBP-3 are involved in apoptosis in the preeclamptic placenta.

Torsion of a parasitic myoma that developed after abdominal myomectomy.

Iatrogenic parasitic myomas are rare. The condition is defined by the presence of multiple smooth-muscle tumorous nodules in the peritoneal cavity. This may be attributable to seeding of myoma particles during uterine surgery. The clinical course is usually indolent. The disease is often asymptomatic and is usually discovered only incidentally. A 38-year-old woman who had undergone abdominal myomectomy 7 months prior presented with acute abdominal pain and a huge pelvic mass. We performed exploratory laparotomy. A parasitic mass 17 cm in diameter with a twisted omental pedicle was identified. En bloc excision of the mass and omentum was performed, followed by total abdominal hysterectomy. Histopathological examination of multiple sections revealed features compatible with an infarcted leiomyoma. Thus, we present a very rare case of an iatrogenic, rapidly growing parasitic myoma complicated by omental torsion (which caused the acute abdominal pain). We also offer a literature review.

Immunologic properties of differentiated and undifferentiated mesenchymal stem cells derived from umbilical cord blood.

The expression of immunogenic markers after differentiation of umbilical cord blood (UCB)-derived mesenchymal stem cells (MSC) has been poorly investigated and requires extensive in vitro and in vivo testing for clinical application. The expression of human leukocyte antigen (HLA) classes on UCB-derived MSC was tested by Fluorescence-activated cell sorting analysis and immunocytochemical staining. The undifferentiated MSC were moderately positive for HLA-ABC, but almost completely negative for HLA-DR. The MSC differentiated to chondrocytes expressed neither HLA-ABC nor HLA-DR. The proliferation of MSC was not significantly affected by the allogeneic lymphocytes stimulated with concanavalin A. The responder lymphocytes showed no significant decrease in proliferation in the presence of the MSC, but the apoptosis rate of the lymphocytes was increased in the presence of MSC. Taken together, these findings indicate that UCB-derived MSC differentiated to chondrocytes expressed less HLA class I and no class II antigens. The MSC showed an immunomodulatory effect on the proliferation and apoptosis of allogeneic lymphocytes. These data suggest that the differentiated and undifferentiated allogeneic MSC derived from umbilical cord blood can be a useful candidate for allogeneic cell therapy and transplantation without a major risk of rejection.

Expression of angiogenic factors in cryopreserved mouse ovaries after heterotopic autotransplantation.

OBJECTIVE: Revascularization is critical for successful ovarian tissue transplantation. Vascular endothelial growth factor (VEGF) and angiopoietin-2 (angpt-2) are the principal mediators of neovascularization. This study was designed to assess VEGF and angpt-2 levels in cryopreserved ovarian tissue after heterotopic autotransplantation. METHODS: Ovarian tissues harvested from ICR mice at 5 to 6 weeks of age were stratified as follows: no cryopreservation (controls, group I); vitrification in VFS-40 (vitrification, group II); and gradual freezing in dimethyl sulfoxide (slow-freezing, group III). Frozen specimens were thawed at room temperature, assaying VEGF and angpt-2 levels 1 week after cryopreservation and 2 weeks after autotransplantation. RESULTS: VEGF and angpt-2 protein levels were significantly lower in cryopreserved ovaries of groups II and III than in controls (group I, P<0.05), whereas groups II and III did not differ significantly in this regard. After autotransplantation of cryopreserved ovarian tissue, VEGF and angpt-2 protein levels did not differ significantly by technique but tended to be lower than corresponding levels in controls. CONCLUSION: Expression of angiogenic factors in ovarian tissue is thought to vary by method of cryopreservation. Our findings indicate that levels of angiogenic factors expressed in cryopreserved ovarian tissue after autotransplantation do not differ appreciably from control levels, regardless of cryopreservation technique.

Influence of the vitrification solution on the angiogenic factors in vitrificated mouse ovarian tissue.

OBJECTIVE: To investigate the effect of the dimethyl sulfoxide (DMSO) and EFS-40 during vitrification on the expression of angiogenic factors in vitrified mouse ovarian tissue. METHODS: The ovarian tissues were obtained from 5 or 6 weeks aged ICR mouse. Ovarian tissues were divided into four groups: ovarian tissue without cryopreservation (control, group I), ovarian tissue vitrified with 15% DMSO (group II), ovarian tissue vitrified with EFS-40 (group III), and ovarian tissue slowly frozen with 10% DMSO (group IV). Thawing was carried out at room temperature. Levels of messenger RNA (mRNA) and protein for vascular endothelial growth factor-A (VEGF-A) and angiopoietin-2 (Angpt-2) were checked in ovarian tissues of four groups recovered on day 7 after cryopreservation. Reverse transcription-polymerase chain reaction and Western blot analysis were used to identify the levels of angiogenic factors in mouse ovarian tissues. RESULTS: Levels of mRNA and protein for VEGF-A and Angpt-2 were significantly decreased in cryopreserved group (group II, III and IV) than control group (group I) (P< 0.05). The significant differences of levels of mRNA and protein for VEGF-A and Angpt-2 between cryopreservation methods were observed (P< 0.05). Group III showed highest expression of mRNA and protein for VEFG-A and Angpt-2 than other cryopreservation groups (P< 0.05). CONCLUSION: These findings suggest that EFS-40 is more efficient vitrification solution for preservation of angiogenic factors than 15% DMSO during vitrification of mouse ovarian tissue. Future studies should investigate to improve the vitrification solution for ovarian tissue vitrification.

DHA and EPA Down-regulate COX-2 Expression through Suppression of NF-kappaB Activity in LPS-treated Human Umbilical Vein Endothelial Cells.

Inflammatory processes of vascular endothelial cells play a key role in the development ofatherosclerosis. We determined the anti-inflammatory effects and mechanisms of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on LPS-treated human umbilical vein endothelial cells (HUVECs) to evaluate their cardioprotective potential. Cells were pretreated with DHA, EPA, or troglitazone prior to activation with LPS. Expression of COX-2, prostaglandin E(2) (PGE(2)) and IL-6 production, and NF-kappaB activity were measured by Western blot, ELISA, and luciferase activity, respectively. Results showed that EPA, DHA, or troglitazone significantly reduced COX-2 expression, NF-kappaB luciferase activity, and PGE(2) and IL-6 production in a dose-dependent fashion. Interestingly, low doses (10 microM) of DHA and EPA, but not troglitozone, significantly increased the activity of NF-kappaB in resting HUVECs. Our study suggests that while DHA, EPA, and troglitazone may be protective on HUVECs under inflammatory conditions in a dose-dependent manner. However there may be some negative effects when the concentrations are abnormally low, even in normal endothelium.

Efficacy of array comparative genomic hybridization in a fetus with an inherited apparently balanced translocation: A case report.

When one parent is discovered to have the same apparently balanced genetic rearrangement as that identified at prenatal diagnosis, many couples are advised to seek a more precise prenatal diagnosis for reassurance. A 25-year-old translocation carrier, who carried a fetus with partial trisomy 3q and partial monosomy 9p with omphalocele in a previous pregnancy, showed the same apparently balanced translocation at chorionic villus sampling. A truly balanced translocation without cryptic imbalances for the fetus was detected using array comparative genomic hybridization. For cryptic unbalanced defects, in which an apparently balanced translocation has been transmitted from a normal parent to a child with a phenotypic abnormality, the use of array comparative genomic hybridization to assess the presence of cryptic aberrations in the fetus combines the speed of DNA analysis with a comprehensive scan for submicroscopic genomic abnormalities.

Expression of clusterin in normal and preeclamptic placentas.

AIM: Clusterin is a multifunctional protein that is up-regulated during many different pathophysiological states. Clusterin may be related to a compensation mechanism involving aggregation of soluble proteins, inhibition of complement-mediated cell damage, adaptive regeneration or apoptosis. Clusterin may increase in response to cell damage that results from hypoxic stress in the preeclamptic placenta. The goal of this study was to measure clusterin expression and localization in normal and preeclamptic placental tissues. METHODS: Immnunohistochemical and double immunofluorescent staining for clusterin was performed on eight normal and eight preeclamptic placental tissues, and clusterin expression was quantified by Western blotting. RESULTS: Immunohistochemical analysis of clusterin in placental tissues obtained from normal pregnancies and those with preeclampsia showed localization of clusterin mainly in the syncytiotrophoblasts and villous endothelial cells. In the double immunofluorescent staining, clusterin was detected in the cytoplasm. The Western blot analysis showed that clusterin expression in the placental tissues of the preeclampsia group was significantly higher than in the normal pregnancy group. CONCLUSIONS: Increased expression of clusterin in placental tissues might play an important role in the pathogenesis of preeclampsia.

Endometrial carcinoma in a patient having 45,X Turner syndrome with gonadal mosaicism.

Only three cases of endometrial carcinoma in women with possibly pure 45,X Turner syndrome without previous unopposed estrogen therapy have been reported. A 46-year-old single nulligravid woman with Turner syndrome phenotype, spontaneous menstruation, and well-differentiated adenocarcinoma of the endometrium was diagnosed as having the 45,X karyotype from peripheral blood, skin, buccal cells, and endometrium, which was confirmed using fluorescence in situ hybridization (FISH). Analysis of the ovarian tissue using FISH confirmed 45,X/46,XX mosaicism. Gonadal mosaicism may help to interpret spontaneous menstruation and endometrial carcinoma in possibly pure 45,X Turner syndrome. We conclude that a molecular analysis of lymphocytes and various tissues is necessary for detecting low-level mosaicism in apparently homogeneous 45,X women.

Effect of vitrification method on survivability, follicular growth and ovulation of preantral follicles in mice.

AIM: This study was designed to compare the survival rates, follicular growth rates, and ovulation rates of vitrified preantral follicles (PF) from ovaries with those isolated from a vitrified ovarian cortical strip. METHODS: Mouse ovaries were divided into three groups: those not treated by vitrification of the PF (control), those treated by vitrification of the PF isolated from the ovaries (group I), and those treated by vitrification of ovarian tissue followed by PF isolation (group II). The group I samples were exposed to equilibration solution (EG-20) for 5.0 min plus vitrification solution (EFS-40) for 0.5 min, while the group II samples were exposed to EG-20 for 10.0 min plus EFS-40 for 2.0 min, before vitrification. They were subsequently placed on an electron microscope grid, and submerged immediately in liquid nitrogen. After thawing, the survival rate and the growth rate of the follicles were evaluated every 2 days. RESULTS: In the in vitro condition, the follicles grew and developed into antral follicles in groups I and II. The survival rate of the group I samples was higher than that of the group II samples during the in vitro culture (P<0.05). The growth rates of the follicles in group I were higher than those in group II after day 6 (P<0.05). The ovulation rate of the samples in group I was higher than that of group II (P<0.05). CONCLUSION: These results demonstrated that direct PF vitrification appeared to be better than vitrification of the PF isolated from ovarian tissue.

Molecular cytogenetic investigation of a balanced complex chromosomal rearrangement carrier ascertained through a neonate with partial trisomies of 13 and 22.

Complex chromosomal rearrangement (CCR) is a structural abnormality of chromosomes that rarely appears in individuals with normal phenotypes. A CCR involving chromosomes 9, 13, and 22 was ascertained in a phenotypically normal woman through a neonate with multiple congenital malformations and partial trisomies of 13 and 22. We diagnosed the CCR using high-resolution chromosome analysis and three-color fluorescence in situ hybridization (three-color FISH) analysis, and ascertained a balanced CCR without cryptic imbalances using array comparative genomic hybridization (array CGH) and FISH. In the present work, we report on the case together with a literature review.

Differential expression of placenta growth factors and their receptors in the normal and pregnancy-induced hypertensive human placentas.

Placental development requires extensive angiogenesis and the invasion of the maternal decidua by the trophoblasts. Adequate and organized interaction of vascular endothelial growth factors (VEGF), placenta growth factors (PlGF), and their receptors are essential for a normal development and function of the placenta. In this study, we evaluated the expressions of PlGFs and their receptors, mRNAs by Northern blotting, in situ hybridization and RT-PCR in the normal and pregnancy-induced hypertensive (PIH) placentas. The expression level of PlGF-2 mRNA was lower in the PIH placentas compared to control as assessed by Northern blotting and in situ hybridization. PlGF mRNA was mainly localized to the vasculosyncytial membrane of placental villi and villous stroma. The expression of PlGF receptor-1 (PlGFR-1) was significantly increased in the PIH placentas compared to the normal ones. These results suggest that the alteration of PlGF-2 and PlGFR-1 mRNA expressions in the placenta are related to the pathogenesis of PIH.