Subpopulations of macrophages within eutopic endometrium of endometriosis patients.

PROBLEM: Endometriosis is recognized as a chronic inflammatory disease and is related to immune response. There have been reports that revealed the different distribution of macrophage within the eutopic endometrium of women with endometriosis. Macrophages are functionally polarized into M1 and M2 cell lineages. We studied a difference in the subpopulations of M1 and M2 macrophages within the eutopic endometrium in patients with or without endometriosis to investigate how the eutopic endometrium is stimulated immunologically. METHOD OF STUDY: Thirty-six patients with endometriosis (endometriosis group) and 37 without endometriosis (non-endometriosis group) were analyzed. Paraffin-embedded endometrial specimens were used for the study. Consecutive sections were used for immunostaining of CD68 (pan-macrophage marker) and CD163 (M2 macrophage marker). Cells positive for each marker were quantified, and the ratio of M2 macrophages in pan-macrophages was calculated. RESULT: The endometriosis group had a significantly higher number of pan-macrophages than the control group in all phases (P < 0.05). The ratios of M2 macrophages in pan-macrophages were significantly lower in all phases in the endometriosis group (P < 0.05). CONCLUSION: The macrophage population slants toward M1 in the endometrium of endometriosis patients. The endometrium appeared to be stimulated by some organelles and/or substances that induce M1 in endometriosis patients.

Progesterone and synthetic progestin, dienogest, induce apoptosis of human primary cultures of adenomyotic stromal cells.

OBJECTIVES: To investigate the direct effects of progesterone receptor (PR) agonists on proliferation and apoptosis of human adenomyotic cells. STUDY DESIGN: Human primary cultures of adenomyotic stromal cells (ASCs) from 24 patients with adenomyosis were co-treated with estradiol (E2) plus the PR agonists, endogenous progesterone (P) or the synthetic progestin dienogest (DNG), which is used to treat endometriosis. In ASCs, anti-proliferative effects and induction of apoptosis were evaluated in the presence or absence of P (10(-8)-10(-6)M) or DNG (10(-8)-10(-6)M) in culture medium containing E2. Cellular proliferation was analyzed with bromodeoxyuridine incorporation and flow cytometry. Apoptosis was detected with annexin V/7-amino-actinomycin D (7-AAD) staining with flow cytometry and cellular caspase 3/7 activity. RESULTS: P and DNG significantly decreased the proportion of cells in the S phase. In addition, both P and DNG increased apoptosis as measured by annexin V-positive/7-AAD -negative cells and caspase 3/7 activity. CONCLUSIONS: Both endogenous P and synthetic progestin directly inhibited cellular proliferation and induced apoptosis in human ASCs. These pharmacological features of progestational compounds provide insight into the therapeutic strategy for the treatment of adenomyosis.

The association between endometriosis and chronic endometritis.

OBJECTIVE: To evaluate the association between endometriosis and chronic endometritis. METHODS: Endometrial specimens were obtained from 71 patients, 34 with endometriosis (endometriosis group) and 37 without endometriosis (non-endometriosis group), who underwent hysterectomy, and the specimens were immunostained for the plasmacyte marker CD138. The rate of chronic endometritis was compared between the endometriosis group and the non-endometriosis group. Furthermore, the 71 patients were also divided into two groups, 28 with chronic endometritis (chronic endometritis group) and 43 without chronic endometritis (non-chronic endometritis group). Logistic regression analysis was performed with variables including age, body mass index (BMI), gravidity and parity, and diagnoses of leiomyoma, adenomyosis, and endometriosis on pathology to examine the independent effect of each variable on chronic endometritis. Patients suffering from cervical invasive carcinoma, endometrial carcinoma, and endometrial polyps or treated with gonadotropin-releasing hormone agonists, progestins, or oral contraceptives before surgery were excluded. RESULTS: Chronic endometritis was identified in 52.94% of the endometriosis group and 27.02% of the non-endometriosis group (p<0.05). Logistic regression analysis revealed that endometriosis was associated with chronic endometritis. CONCLUSIONS: This result suggests a strong association between endometriosis and chronic endometritis.

Four subtypes of adenomyosis assessed by magnetic resonance imaging and their specification.

OBJECTIVE: The aim of the present study was to differentiate and specify the subtypes of adenomyosis. STUDY DESIGN: Surgically treated adenomyosis (n = 152) was subcategorized retrospectively into 4 subtypes on the basis of magnetic resonance imaging geography. Subtype I (n = 59) consisted of adenomyosis that occurs in the uterine inner layer without affecting the outer structures. Subtype II (n = 51) consisted of adenomyosis that occurs in the uterine outer layer without affecting the inner structures. Subtype III (n = 22) consisted of adenomyosis that occurs solitarily without relationship to structural components. Adenomyosis that did not satisfy these criteria composed subtype IV (n = 20). Stepwise logistic regression analysis was used for specification of the subtypes. RESULTS: Subtypes I-III were suggested as a product of direct endometrial invasion, endometriotic invasion from the outside, and de novo metaplasia, respectively. Subtype IV was a heterogeneous mixture of far advanced disease. CONCLUSION: Adenomyosis appears to consist of 3 distinct subtypes of different causes and an additional subtype of indeterminate cause.

Possible involvement of the E-cadherin gene in genetic susceptibility to endometriosis.

BACKGROUND: Endometriotic cells display invasive characteristics, despite their benign histological appearance. Recently, the epithelial-mesenchymal transition, in which epithelial cells acquire mesenchymal and migratory properties, has attracted attention as a mechanism of tumor invasion. We aimed to investigate the association between endometriosis and polymorphisms of the E-cadherin gene, a central player in the epithelial-mesenchymal transition, in Japanese women. METHODS: Twelve single-nucleotide polymorphisms (SNPs) in the E-cadherin gene were identified by real-time polymerase chain reaction using a TaqMan assay in 511 women with endometriosis (the majority in Stages III and IV) and 498 healthy controls. RESULTS: Allele frequency analysis indicated that there was a marginally higher frequency of the rs4783689 C allele in women with endometriosis compared with controls (corrected P = 0.007; odds ratio = 1.37; 95% confidence interval, 1.14-1.64). No significant associations with endometriosis were found for the other 11 SNPs. CONCLUSIONS: Although this study was limited by sample size, the E-cadherin gene polymorphism rs4783689 was marginally associated with endometriosis in the Japanese population, suggesting that E-cadherin might be involved in genetic susceptibility to endometriosis.

Meta-analysis of genome-wide association scans for genetic susceptibility to endometriosis in Japanese population.

To identify susceptibility genes for endometriosis in Japanese women, genome-wide association (GWA) analysis was performed using two case-control cohorts genotyped with the Affymetrix Mapping 500K Array or Genome-Wide Human SNP Array 6.0. In each of the two array cohorts, stringent quality control (QC) filters were applied to newly obtained genotype data, together with previously analyzed data from the Japanese Integrated Database Project. After QC-based filtering of samples and single nucleotide polymorphisms (SNPs) in each cohort, 282 838 SNPs in both genotyping platforms were tested for association with endometriosis using a meta-analysis of the two GWA studies with 696 patients with endometriosis and 825 controls. The meta-analysis revealed that a common susceptibility locus conferring a large effect on the disease risk was unlikely. On the other hand, an excess of SNPs with P-values <10(-4) (36 vs 28 SNPs expected by chance) was observed in the meta-analysis. Of note, four of the top five SNPs with P-values <10(-5) were located in and around IL1A (interleukin 1α), which might be a functional candidate gene for endometriosis. Further studies with larger case-control cohorts will be necessary to elucidate the genetic risk factors.

Prevention of postlaparoscopic shoulder pain by forced evacuation of residual CO(2).

BACKGROUND AND OBJECTIVES: Shoulder pain is one of the early postlaparoscopic symptoms related to CO(2) used for pneumoperitoneum and remaining in the abdomen. The present study was conducted to validate the hypothesis that complete evacuation of the residual CO(2) would prevent postlaparoscopic shoulder pain. METHODS: Forty consecutive patients, the candidates for gynecologic laparoscopic surgery, were randomly enrolled into one of the following 2 groups. Nineteen patients entered Group I where the residual CO(2) was evacuated by abdominal oppression and served as the study control group. The remaining 21 patients entered Group II, where the residual CO(2) was evacuated by pumping warm saline into the abdomen until it spilled out of the open ports. Nurses, blind to the patient's grouping, recorded shoulder pain VAS scores twice daily. RESULTS: VAS scores in Group I started to increase at Day 1AM, reached a peak at Day 1PM, and decreased gradually thereafter. VAS scores in Group II stayed low throughout the investigation period. The difference was highly significant (P<0.001). CONCLUSIONS: Abdominal filling with saline at the end of laparoscopic surgery effectively evacuates residual CO(2) thus preventing postlaparoscopic shoulder pain.

Two novel serum biomarkers for endometriosis screened by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry and their change after laparoscopic removal of endometriosis.

OBJECTIVE: To explore novel endometriosis serum biomarkers by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). DESIGN: First, we aimed to discover the potential biomarkers of endometriosis by SELDI-TOF-MS. Second, blinded test was performed to characterize the effectiveness of the model by examining the sensitivity and specificity. Third, 29 postoperative patients with endometriosis were recruited to monitor the change of potential biomarkers after laparoscopic surgery. SETTING: Collaborative investigation in an academic research environment. PATIENT(S): Fifty-nine patients with endometriosis, 31 patients without endometriosis, and 30 healthy volunteers. INTERVENTION(S): Blood serum of endometriosis and control group patients. MAIN OUTCOME MEASURE(S): Protein expression. RESULT(S): Two endometriosis-specific proteins were found in the preliminary screening study. Furthermore, the blinded test was performed and showed a sensitivity of 86.67% and a specificity of 96.77% of the markers for detecting endometriosis, which are significantly higher than those of CA-125 for distinguishing patients with endometriosis from patients without endometriosis. After surgery, the levels of these biomarker proteins decreased to levels comparable with those of patients without endometriosis. CONCLUSION(S): We discovered the potential biomarkers of endometriosis and set up a diagnostic model with a sensitivity of 86.67% and a specificity of 96.77%, which is significantly higher than that of CA-125 for detecting endometriosis, The levels of these proteins decreased to levels comparable with those of patients without endometriosis.

Unusual uterus didelphys presenting in the retroperitoneum.

We describe a case of uterus didelphys with obstructed hemivagina and ipsilateral renal agenesis. Magnetic resonance imaging (MRI) demonstrated uterus didelphys and laparoscopy revealed that an affected uterus was present in the retroperitoneum.

Microencapsulated feeder cells as a source of soluble factors for expansion of CD34(+) hematopoietic stem cells.

Expansion of hematopoietic stem cells (HSCs) from cord blood is highly desired for treatment and transplantation of adult patients for hematologic diseases. For efficient proliferation of HSCs, CD34(+) cells from cord blood were co-cultured with microencapsulated murine stromal cells (HESS-5) or immortalized human mesenchymal stem cells (MSCs) in their conditioned media (CM). Bioactive substances for HSC proliferation in CM at the onset of culture are likely consumed by HSCs with time, and co-culturing with microencapsulated feeder cells ensures a continuous supply. The cell number of CD34(+) cell progeny efficiently increased under these culture conditions, and progeny were analyzed by flow cytometry, the colony assay and the cobblestone area-forming cell (CAFC) assay. Total nucleated cells and CD34(+) cell number increased 194- and 7.4-fold, respectively, in the presence of microencapsulated HESS-5 in CM. Colony forming cells and CAFCs were well maintained. The effective expansion of total cells and maintenance of primitive progenitor cells suggest that transfusion of the progeny obtained from CD34(+) cell culture with microencapsulated HESS-5 in CM could shorten the time to engraftment by bridging the pancytopenic period and support functional hematopoietic repopulation.

Use of the potassium titanyl phosphate laser in the laparoscopic anterior rectum slicing operation.

OBJECTIVE: To evaluate the efficacy of the potassium titanyl phosphate (KTP) laser used during the laparoscopic anterior rectum slicing (LARS) operation. DESIGN: Retrospective study. SETTING: National Hospital Organization Kyoto Medical Center, Kyoto, Japan. PATIENT(S): All 46 patients who underwent the LARS operation using the KTP laser. INTERVENTION(S): The LARS operation using the KTP laser for treatment of deep rectal endometriosis. MAIN OUTCOME MEASURE(S): Operative and postoperative outcome. RESULT(S): Meaningful improvements in clinical symptoms were obtained with the LARS operation using the KTP laser with acceptable levels of postoperative morbidity. Bowel leakage did not occur in any of the patients. CONCLUSION(S): Deep rectal endometriosis can be treated effectively with the LARS operation using the KTP laser.

Complete removal of endometriosis improves fecundity.

Endometriosis is one of the causative factors of impaired fecundity. Whatever the mechanisms are of this impairment, surgical removal of endometriosis appears to increase postoperative fecundity. Our strategy in laparoscopic surgery for symptomatic endometriosis is to completely remove endometriosis. However, despite this strategy, laparoscopic surgery nonetheless creates two categories of patients; complete and incomplete surgery groups. We found by comparing the two groups that both were comparable in terms of fecundity during the early postoperative phase, whereas unlike the complete surgery group, fecundity in the incomplete surgery group stayed low during the late postoperative phase. Deep rectal endometriosis and deeply invading pelvic endometriosis are conditions wherein complete removal of endometriosis is difficult. We have developed laparoscopic surgeries for these conditions: laparoscopic anterior rectum slicing (LARS) and laparoscopic pelvic wall slicing (LPWS) operations, respectively. Both operations are effective in alleviating disease-related symptoms with minimal surgical invasiveness.