Neoplasms

Urinary Tract, Sexual Organs, and Pregnancy Conditions

Gland and Hormone Related Diseases

All Conditions

Symptoms and General Pathology

Wounds and Injuries

Rare Diseases

Ovarian Neoplasms

Hyperthermia

Endocrine Gland Neoplasms

Ovarian Diseases

Adnexal Diseases

Genital Diseases, Female

Genital Diseases

Urogenital Diseases

Female Urogenital Diseases

Pregnancy Complications

Female Urogenital Diseases and Pregnancy Complications

Genital Neoplasms, Female

Urogenital Neoplasms

Endocrine System Diseases

Gonadal Disorders

Ovarian Cancer

Coagulants

All Drugs and Chemicals

Hemostatics

After laparoscopic ovarian cystectomy, hemostasis will be performed using FloSeal (1 vial of FloSeal 5mL per unilateral ovarian cystectomy)

After laparoscopic ovarian cystectomy, hemostasis will be performed using electrocautery

FloSeal application

Electrocautery

Joo-Hyun Nam, M.D., Ph.D.

To compare the efficacy in postoperative hemostasis and in sparing postoperative ovarian function between FloSeal and Electrocauterization in laparoscopic ovarian cystectomy

Efficacy Study of FloSeal in Hemostasis After Laparoscopic Ovarian Cystectomy

PROSPECTIVE RANDOMIZED CONPARISON OF ELECTRONIC DIATHERMY VERSUS FLOSEAL IN HEMOSTASIS AFTER LAPAROSCOPIC OVARIAN CYSTECTOMY

Postoperative complications will be evaluated with Dindo's classification for postoperative complication and CTCAE v4.0.

Baxter Healthcare Corporation

Asan Medical Center

Professor

Carcinoma, Ovarian Epithelial

Infertility

Ovarian Epithelial Cancer

25 patients with benign ovarian tumor (cystadenoma) were admitted to IInd Department of Gynecology, Lublin Medical University, Lublin, Poland.

11 women with borderline ovarian tumor were admitted to IInd Department of Gynecology

24 women with ovarian cancer were admitted to IInd Department of Gynecology

20 patient with unexpleined infertility were admitted to IInd Department of Gynecology

Treg, Th17, NKT cells in peripheral blood and ovarian tissue and serum level of HE4, CA125 and ROMA.

Assessment of percentage of Treg, Th17, NKT and serum level of HE4, CA125 and ROMA.

Izabela Winkler

The aim of the study was to estimate the percentage and of Treg, Th17 and NKT in peripheral blood and the tissue of the epithelial ovarian tumor and relationship with blood serum level of HE4, CA125, as well as algorithm ROMA.

Material and methods Mononuclear cells (PBMCs) were isolated by density gradient centrifugation obtained from peripheral blood and ovarian tissue of patient suffering ovarian pathology. Patient from control group underwent surgery for unexplanied infertility. The percentage of Treg and Th17 , NKT in peripheral blood and the tissue was assessed using the flow cytometry method according to the manufacturer's instructions. The ROMA index was calculated according to the levels of HE4 and CA-125 in serum.

Study groups. The study group consisted of 60 women. The patients were divided into 3 subgroups: a group of 24 women with malignant epithelial ovarian tumors (cystadenocarcinomas), 25 women with benign ovarian tumors (cystadenomas) and 11 women with borderline ovarian tumors (serous borderline tumors). The control group consisted of 20 women without ovarian pathology who underwent surgery for unexplanied infertility. Patients were admitted to IInd Department of Gynecology, Lublin Medical University, Lublin, Poland, between 2011 and 2014. All women with OCs were staged III or IV according to revised 2013 FIGO classification (International Journal of Gynecology and Obstetrics, January, 2014) Isolation of mononuclear cells from peripheral blood (PBMCs). Immediately after being taken from the antecubital vein, mononuclear cells (PBMCs) were isolated by density gradient centrifugation applying Gradisol L formulation of specific gravity 1.077g/ml (Aqua Medica, Łódź, Poland) for 20 min. at 700 x g. The pellet containing PBMC was washed twice in PBS and evaluated for the number (using Neubauer chamber) and viability (trypan blue staining - 0.4% trypan Blue Solution, Sigma-Aldrich, Munich, Germany). Viability of less than 95% was a disqualifying criterion.

Isolation of mononuclear cells infiltrating the tumor and healthy tissues.

During the surgery, ovarian tissue (size of approximately 1cm3) not containing any visible necrotic areas was collected. It was minced with a scalpel, suspended in 30 ml of RPMI 1640 medium (Biochrome, Holliston MA, USA) and subjected to cleavage in a mixture containing: 1 mg/ml collagenase type IA (Sigma-Aldrich, Munich, Germany), 1 mg/ml DNase type I (Sigma-Aldrich, Munich, Germany), 0.1 mg/ml hyaluronidase (Sigma-Aldrich, Munich, Germany) at 37°C for 60 min., constantly vortexed. After cleavage, the suspension was filtered through a strainer (70μm, BD Biosciences, San Jose CA, USA) and centrifuged for 5 min. at 700 x g. The suspension cells were washed twice in RPMI 1640.

Evaluation of the percentage of Th17 cells.

The evaluation of the percentage of Th17 cells (secreting IL-17A) of peripheral blood mononuclear cells, healthy tissue and tumor tissue was performed by flow cytometry using a Th17 Cytokine Staining panel according to the manufacturer's recommendation (eBiosciences, San Diego, USA) using a FACSCanto (BD Biosciences, San Jose CA,USA).

Establishment of PBMC culture and ovarian tissue (tumor or control) and stimulation with ionomycin.

After 24-hour culture of the PBMCs and ovarian tissue was set up. The medium was prepared consisting of 97% RPMI 1640 (Biochrome, Holliston MA,USA) supplemented with 2mM L-glutamine, 2% human albumin (ZLB Bioplasma, Bern, Switzerland) and antibiotics in an amount of 100 U/ml penicillin (Sigma-Aldrich, Munich, Germany) and 100 mg/ml streptomycin (Sigma-Aldrich, Munich, Germany). Cultivation was carried out in 6-well plates in a 5 ml culture medium. For each patient, two cultures were established, one of the PBMC and the other from tumor cells or normal ovarian tissue. The culture was conducted in an incubator under standard conditions (5% CO2, 95% humidity, 37°C) for 4 hours. To individual wells were added: ionomycin (Sigma-Aldrich, Munich, Germany) at a concentration of 1 ug/ml and PMA at a concentration of 25 ng/ml to stimulate cells for the production of cytokines and brefeldin at a concentration 10 μg/ml (Sigma-Aldrich, Munich, Germany) in order to inhibit the activity of the endoplasmic reticulum, leading to retain the cytokine within the cell.

The determination of intracellular cytokines.

24-hour cultures after moving from the culture plate to two properly signed tubes were washed twice in 2 ml of Flow Cytometry Staining Buffer (eBioscience, San Diego, USA) after vortexing. Constant parameters used during each rinsing in this procedure are: run time 5 min., 700 x g. After removing the supernatant, 5μl of anti-CD4 (eFluor 450, eBioscience, San Diego, USA) was added to each tube. The mixture was incubated for 20 min. in darkness. After washing away, the excess of antibody in 2 ml of Flow Cytometry Staining Buffer (eBioscience, San Diego,USA), 100μl IC Fixation Buffer (eBioscience, San Diego, USA) was added to each tube in order to consolidate. Mixing/Vortexing this mixture was also incubated for 20 minutes in darkness. Then it was washed twice with 2 ml of the permeabilization buffer (Permeabilization Buffer, eBioscience, San Diego, USA). Preparing the buffer involved its 10-fold dilution with PBS. After removing the supernatant, the cells were resuspended in 100μl of permeabilization buffer and separated to previously prepared cytometry tubes into control and test samples. Thereafter, 5μl antibodies were added to the test samples: anti-IL-17A (FITC). After vortexing, all samples were incubated for 20 min. in darkness. They were washed twice and after suspending the cells in the Flow Cytometry Staining Buffer, cytometric analysis was carried out. Analysis was performed using flow cytometric BD FACSCanto II (BD Biosciences, San Diego, USA). Measurements were performed using the software BDFACS Diva.

Estimation of the percentage of regulatory T lymphocytes.

Estimation of the percentage of regulatory T lymphocytes among peripheral blood mononuclear cells and in healthy and neoplastic tissue was made with the flow cytometry method using the Human Treg Flow™ Kit (FOXP3 Alexa Fluor® 488 / CD4 PE-Cy5/CD25 PE, BioLegend®, USA) with the use of FACSCanto apparatus (BD Biosciences, San Diego,USA).

Determination of iNKT cell subpopulation by flow cytometry

Mononuclear cells isolated from peripheral blood, healthy tissue and epithelial ovarian tumors were incubated with the appropriate volume (according to the procedures recommended by the manufacturers) of the appropriate monoclonal antibodies: the anti-iNKT FITC (anti-Vα24) (cat#558371, clone 6B11, BD Pharmingen, San Jose, CA, USA), anti-CD3 PE (Cat #554829, clone G4. 18, BD Pharmingen, San Jose, CA, USA) and anti CD161-PE f(clone Dx12, Cat#550968, BD Pharmingen, San Jose, CA, USA). Prepared samples were incubated for 20 min. at room temperature in the dark. After twice washing in buffered physiological saline (PBS), the cells were immediately subjected to cytometric analysis. Data acquisition and analysis were performed using a FACS Calibur flow cytometer (Becton Dickinson, New Jersey, USA). Expressing cells iNKT+/CD3+/CD161+ were rated among CD3+

Assessment of concentration of protein HE4, CA125 and algorithm ROMA.

Samples were collected from all the patients prior to surgery and 3 ml blood was collected. Serum was centrifuged at 2000 × g and stored at -20 and -80°C until use. Serum level of CA-125 and HE4 were detected using the full automatic chemiluminescence analyzer Cobs601 and the test procedure was performed according to the manufacturer's instructions (Roche Diagnostics, Indianapolis, IN, USA). Next, serum HE4 and CA-125 levels were calculated for ROMA index value using the Roche ROMA index of ovarian cancer risk assessment software. Serum HE4 and CA-125 reference range was \<140 pmol/l and \<32 U/ml, respectively.

Assessment of risk of epithelial ovarian carcinoma (ROMA) The ROMA index was calculated according to the levels of HE4 and CA-125. HE4 and CA-125 amount were input to the ovarian cancer risk assessment software, followed by automatic calculation of the corresponding ROMA index. The premenopausal calculation formula of the ROMA index was: 12+2.38 × LN(HE4)+0.062 6 × LN(CA-125). The postmenopausal calculation formula of the ROMA index was: 8.09+1.04 × LN(HE4)+0.732 × LN(CA-125). Premenopausal women with a ROMA value ≥11.4, had a higher risk of ovarian cancer. Postmenopausal women with ROMA value ≥29.9 had a higher risk of ovarian cancer.

Treg, Th17 Cells, NKT in Epithelial Ovarian Tumor

Lymphocyte T Regulatory , Th17 and NKT in Epithelial Ovarian Tumor- Prognostic Assessment and Relationship With Clinical Marker

Percentage of regulatory TREG, Th17, NKT among peripheral blood mononuclear cells and in healthy and neoplastic tissue was made with the flow cytometry

Assessment level of CA125 and HE4 in serum

Kaplan-Meier survival analysis was performed in the group of patients with ovarian cancer.

Spearman's rank correlation coefficient and its significance test was applied to assess relationships between two parameters.

Medical University of Lublin

Clinical Proffesor

Carcinoma

Neoplasms, Glandular and Epithelial

Neoplasms by Histologic Type

Methylation testing of host DNA, namely, OPCML, FODX3 and CDH13, in the peripheral serum

DNA methylation testing

Lei Li, M.D.

Ming Wu, M.D.

Lei Li, MD

Liquid biopsy is challenging for the diagnosis of epithelial ovarian cancer (EOC). In this study, we performed the methylation testing of host DNA, namely, OPCML, FODX3 and CDH13, in the peripheral serum to discover the diagnostic and supervision roles of DNA methylation in EOC patients. The study compromises two stages.

In the training set, DNA methylation testing is performed in the ovarian tissues from EOC and paired benign ovarian tumor patients. The cut-off values of methylation are produced in this stage. On the meantime, serum DNA methylation testing is also performed to reveal its accordance and accuracy compared with the results of ovarian tissues.

In the validation set, serum DNA methylation testing is performed in unselected ovarian tumor patients with definite cut-off values to validate its accuracy based on known histology of ovarian tumors.

In training and validation sets, serum DNA methylation is also performed after major surgeries for EOC as to illustrate the changes of methylation testing, therefore, reflection the supervision role of DNA methylation.

All eligible patients accept DNA methylation testing in serum and in ovarian tissues.

Cell-free DNA Methylation for Epithelial Ovarian Cancer

Cell-free DNA Methylation for the Diagnosis and Monitoring of Epithelial Ovarian Cancer: The Training and Validation Sets in China

The cut-off values are achieve by paired epithelial ovarian cancer and benign ovarian tumor patients.

The accuracy is supported by sensitivity, specificity, negative predictive value and positive predictive value.

Study title	NCT ID	Conditions	Interventions	Eligibility	Organization
Efficacy Study of FloSeal in Hemostasis After Laparoscopic Ovarian Cystectomy	NCT01683877	Benign Ovarian ...	FloSeal applica... Electrocautery	20 Years -	Asan Medical Center
Treg, Th17 Cells, NKT in Epithelial Ovarian Tumor	NCT03779399	Ovarian Cancer Borderline Ovar... Benign Ovarian ... Unexplained Inf...	Assessment of p...	18 Years - 75 Years	Medical University of Lublin
Cell-free DNA Methylation for Epithelial Ovarian Cancer	NCT04651946	Epithelial Ovar... DNA Methylation Benign Ovarian ... Training Set Validation Set Cell-free DNA Liquid Biopsy	DNA methylation...	18 Years -	Peking Union Medical College Hospital
Cell-free DNA Methylation for Epithelial Ovarian Cancer	NCT04651946	Epithelial Ovar... DNA Methylation Benign Ovarian ... Training Set Validation Set Cell-free DNA Liquid Biopsy	DNA methylation...	18 Years -	Peking Union Medical College Hospital
Efficacy Study of FloSeal in Hemostasis After Laparoscopic Ovarian Cystectomy	NCT01683877	Benign Ovarian ...	FloSeal applica... Electrocautery	20 Years -	Asan Medical Center

Spots Global Cancer Trial Database for benign ovarian tumor