Objective To establish the high-performance liquid chromatography(HPLC)method for measuring concentrations of methotrexate(MTX)in rat lung and some other tissues through internal iliac artery infusion.Methods Fifly female Sprague-Dawley rats were included in this study.The rats were randomly assigned to two groups.Methotrexate was injected to group one through internal iliac artery,and was injected to group two through femoral vein.Blood and tissues were collected in each group at 15,30,60.90 and 120 minutes for detection of the drug concentrations with HPLC.Results The area under the concentration time curve(AUC)in rat lung,ovary and uterus in the artery group were separately(3.77±0.28),(4.40±0.40),(9.97±0.89)μg·h-1·g-1,which were significantly different from those of the vein group[(2.31±0.25),(3.91±0.19),(7.65±1.54)μg·h-1·g-1;P<0.05].The AUC in the rat plasma,heart,kidney,liver and spleen in the artery group were separately(6.13±0.53),(1.90 ±0.11),(5.32±0.89),(14.16±1.96),(0.76±0.20)μg·h-1·g-1.There were no significant differences from the vein group[(5.79±0.71),(1.64±0.29),(5.15±1.69),(14.29 ±3.47),(0.76±0.13)μg·h-1·g-1;P>0.05].Conclusions Through internal iliac artery infusion,there are higher drug concentrations in lung.uterus and ovarian compared to venous injection.The internal-arterial chemotherapy may be used to treat pulmonary metastasis of gynecological tumor.

Objective To establish a satisfactory lung metastasis model of human choriocarcinoma using severe combined immunedeficient(SCID)mice and explore the appropriate cell concentration for the model.Methods Forty SCID mice aged between 5-6 weeks were randomly difided into four groups.1×107 cells/ml ×0.1 ml.5×106 cells/ml×0.2 ml and 1×106 cells/ml×0.1 ml of human choriocarcinoma cells JEG-3 were respectively injected in SCID mice of experimental groups by lateral tail vein,the remain group was assigned to the control group.The status and weisht of mice were observed every three days.When these mice were being dying.the size and the number of the lesions of lung metastasis in every mouse were inspected with Micro CT.After Micro CT inspection,the SCID mice were executed dissected to note whether there were tumors on all organ surfaces witll naked eyes.then made pathological sections from the metastaticfoci of fresh lung tissues,and cultured primarily cells and purified cells and passaged cells isolated from the same metastastic foci.The pathological sections were observed under the microscope.The special antigen human chorionic gonadotropin-beta subunit(β-hCG)of the choriocareinoma cells was immunohistochemically detected in the pathological sections and the cells out of cultured primarily cells.The chromosomes of the cells out of cultured primarily cells were analysed.Results Of the group inoeutated 1×107 cells/ml×0.1 ml.all mice died when inoculating.In the group of 5×106 cells/ml×0.2 ml,when inoculating, 3 mice died; the remain 7 mice were being dying on ( 18. 0 ±2. 0) days after injection. 5 of them, there were 1 - 3 lesions of lung metastasis after Micro CT inspection in each mice, and the diameter of the tumors lesions reached 1.5 - 3.5 ram, which was choriocarcinoma confirmed by pathological sections.The special antigen β-hCG was detected by immunohistoehemical method in the pathological sections of pulmonary tissue with tumor and in the cells, which were purified and passaged from being cultured primarily cells isolated from metastastic foci of fresh lung tissues from the SCID mice. The chromosome numbers of these cells out of cultured primarily ceils were variety from 19 to 128, and medal numbers were variety from 70 to 79. Conclusions We successfully established the lung metastatic model of human choriocarcinoma in SCID mice by injecting JEG-3 cells into lateral tail vein, of which 5 × 106 cells/ml × 0. 2 ml is the suitable concentration and volume for the model.

Objective To evaluate the feasibility and safety of transabdominal ultrasound-guided suction curettage to treat cesarean scar pregnancy (CSP) and investigate factors affecting prognosis of CSP. Methods This was a retrospective case study of 43 cases who were diagnosed as CSP and treated by suction curettage under transabdominal ultrasound guidance as an initial treatment of CSP in Obstetrics and Gynecology Hospital of Fudan University over past 7 years (from 2007 to 2013);factors affecting prognosis of CSP were investigated. Results 39 of the 43 cases (91%) were successfully treated. There were no statistically significant differences in maternal age, gravidity, abortion frequency, and the time interval between current CSP and last cesarean delivery, the myometrium thickness between the gestational sac and the bladder wall between the success group and the failure group (all P>0.05). Statistically significant difference was found in crown-rump length (CRL) between the two group (median of the two group was 18.5, 2.0 mm) by rank sum test (P=0.047). Univariate logistic regression analysis demonstrated that CRL was strongly associated with the prognosis and the OR for no complications was 18.50, comparing CRL≤6 mm versus CRL>6 mm (P=0.020). Conclusion Transabdominal ultrasound-guided suction curettage is effective and safe in the treatment of CSP with CRL≤6 mm.

Objective To investigate gene transfer efficiency of a novel target non-viral vector GE7 and effects of herpes simplex virus thymidine kinase (HSV 1-tk)/ganciclovir(GCV) mediated by it in vitro. Methods The epidermal growth factor receptor (EGF-R) target gene delivery system GE7 was constructed.Human ovarian cancer cell line CAOV3 was transfected in vitro with ?-galactosidase(?-gal) as reporter gene and HSV 1-tk gene as therapeutic gene using this gene delivery system.By means of the assay of X-gal staining, Northern blotting, cell growth-inhibiting curve and so on,the transferring efficiency of exogenous genes and killing effects are observed. Results It showed that gene transfer efficiency is over 80%.When 10 mg/L GCV was put into ovarian cells transfected with HSV 1-tk gene, 95% of cells were killed, and the apoptosis ratio reached up to 30. Conclusions The GE7 gene delivery system is an effective and safe delivery system.GE7/ HSV 1-tk /GCV therapeutic gene system is appraising for ovarian cancer.

Objective To investigate the efficiency of antitumor immune responses induced by a controlled live dendritic cell(DC)vaccine Methods DC precursors were isolated from Fischer 344 rat bone marrow and cultured with granulocyte macrophage colony stimulating factor and interleukin 4 The rat ovarian tumor cell line NuTu 19 was genetically modified by retroviral mediated suicide gene(HSV 1 TK), and the positive clones were selected using G418 Live DC vaccine was then fused with DC and NuTu 19/TK cell by polyethylene glycol The characteristics of live DC vaccine were assayed with flow cytometry and confocal laser scanning microscopy The specific expression of HSV 1 TK gene in live DC vaccine was evaluated by RT PCR and western blot The sensitivity of live DC vaccine to ganciclovir (GCV) was evaluated by methylthiazoletetrazolium assay In vivo, rats vaccinated twice with live DC vaccine were compared to those vaccinated with killed DC vaccine, unfused DC and NuTu 19/TK cell or phosphate buffered saline Seven days following the last immunization, the rats were sacrificed to test the specific cytotoxic T lymphocyte (CTL) activity by lactate dehydrogenase release assay, or challenged with NuTu 19 and tumor incidence was observed Results The fusion efficiency was approximately (23?14) Live DC vaccine displayed an up regulated expression of major histocompatibility complex (MHC) IIOX6 (87 6?3 4)%, costimulatory molecule B 1 2 (71 1?9 3)%, integrin OX 62 (68 0?7 4)%, and adhesion ICAM 1 (77 1?2 0)%, and specifically expressed HSV 1 TK gene. Our data showed that spleen T lymphocytes from rats vaccinated with live vaccine displayed enhanced CTL aetivity (61 8?8 3)% contrast to that of rats vaccinated with killed vaccines (26 0?3 8)% ( P

Objective To investigate the expression of exogenous gene transferred by piggyBac (PB) transposon in various gynecological malignant cell lines and reveal its potential application of gene therapy in gynecological cancer.Methods Amplified herpes simplex virus thymidine kinase (HSV-tk) gene coding region by PCR and integrated it into PB expression vector, PB[Act-RFP]DS, for reconstructing PB[Act-RFP, HSV-tk]DS (pPB/TK).By using different transfection reagents: FuGENE HD, jetPEI, lipofectamine 2000, pPB/TK together with helper plasmid Act-PBase were cotransfected into four mostly common gynecological malignant tumor cell lines HeLa, JEG-3, SKOV3 and HEC-1B.The mRFP1 report gene expressions was observed and detected by fluorescence microscope and flow cytometry to analyze transfection efficiency.The expressions of HSV-tk and mRFP1 gene were detected by reverse transcription PCR (RT-PCR).The cytotoxic effect of various concentration of pro-drug ganciclovir (GCV) for transfected cells was detected by methyl thiazole tetrazolium assay.The transfected cells were positive sorted by flow cytometry and limiting diluted to obtain the stable transfected cell line.The insertion sites of foreign gene tranferred by PB transposon in genome were analyzed by inverse PCR.Results (1) Double digests analysis and sequences test demonstrated that pPB/TK vector was reconstructed successfully.(2) Using three different transfective reagents, PB trausposon transferred HSV-tk gene and mRFP1 gene into HeLa, HEC-1 B, SKOV3 and JEG-3 cell efficiently, and the transfection efficiency of pPB/TK for the same cell was different by using different transfective reagents; in Hela cell, the transfection efficiency of FuGENE HD [(78.7 ± 9.2) %]was higher than that of lipofectamine 2000 [(54.1 ± 11.4) %]and jetPEI [(46.5 ± 7.4) %, all P < 0.05] ; using the same transfective reagent, the transfection efficiency of pPB/TK was also different on various cell lines, using FuGENE HD, the transfeetion efficiency of pPB/TK on HeLa, JEG-3 and SKOV3 cell was (78.7 ± 9.2) %, (74.4 ± 8.9) % and (83.2 ± 9.7) % respectively, which all were higher than that on HEC-1B [(39.5 ± 8.7) %, P < 0.05] .(3) RT-PCR showed that there were the mRNA expression of HSV-tk and mRFP1 in all cell lines.(4) 50% inhibitory concentration of GCV for transfected cells, HeLa, JEG-3, SKOV3 and HEC-1B, was 1.29, 3.35, 0.09 and 13.28 μg/ml respectively.Inhibitory effect of GCV (10 μg/ml) on SKOV3 transfected with pPB/TK was (86 ± 9) %, which was superior to that transfected with pORF-HSVtk alone [(52 ± 12)%, P < 0.05] .(5) The insertion sites of PB transposon in the target cells genome were located at TTAA sites, mRFP1 expression still could be detected in three months after transfected.Conclusions PB transposon could transfer exogenous gene into various gynecological malignant cells, which could integrated into genome and obtain a long-term and stable expression.It is expected that PB transposon may supply a more efficient and safer transgene technology platform for gene therapy in gynecological cancer.

OBJECTIVE: Poly (ADP-ribose) polymerase (PARP) is an important molecule in the early stress response of DNA damage, which is involved in DNA damage repair and cellular senescence. Olaparib, as PARP inhibitor, has an anti-tumor effect on high grade serous ovarian cancer, but its effects on cellular senescence have not been reported. This study intends to explore the role of olaparib in the regulation of senescence in ovarian cancer cells. METHODS: The effects of olaparib on the senescence of ovarian cancer cells were detected by using the senescence-associated β-galactosidase (SA-β-Gal) and senescence-associated heterochromatin aggregation (SAHF). Quantitative real-time polymerase chain reaction was used to analyze the senescence-associated secretory phenotype (SASP). Cell cycle and apoptosis were detected by flow cytometry. The effect of olaparib on tumor growth was analyzed in a nude mouse xenograft transplantation model. RESULTS: Long-term (6 days) treatment with olaparib (5 μM) significantly inhibited the growth of ovarian cancer cells, leading to arrest the cell cycle at G0/G1 phase, significant increase the number of positive SA-β-Gal stained cells and positive SAHF cells. The expression of P16 and retinoblastoma protein (p-RB) were significantly enhanced in SKOV3 cells under olaparib treated, meanwhile, the expression of P53 and p-RB were upregulated in A2780 cells. In OVCAR-3 cells, the expression of P53 was downregulated and p-RB was upregulated. Mice with SKOV3 xenograft transplantation was given olaparib (10 mg/kg/day) via abdominal cavity administration, the tumor volume was reduced (p < 0.01). CONCLUSION: Continuous low dosage administration of olaparib induced senescence under P16 or P53 dependent manner in ovarian cancer.
Abdominal Cavity ; Aging ; Animals ; Apoptosis ; Cell Aging ; Cell Cycle ; DNA Damage ; Flow Cytometry ; Heterochromatin ; Mice ; Mice, Nude ; Ovarian Neoplasms ; Phenotype ; Real-Time Polymerase Chain Reaction ; Retinoblastoma Protein ; Transplantation, Heterologous ; Tumor Burden

Non-peptide gonadotropin-releasing hormone antagonist(GnRH-antagonist)is a highly efficient and selective antagonist of GnRH receptor.The oral preparations(Elagolix,Relugolix and Linza-golix)had been approved for listing abroad.It shows efficacy in reducing the size of uterine fi-broid and controlling heavy bleeding.Appropriate combination of'add-back'therapy could improve the safety in medical application and patient com-pliance.Compared with other hormonal pharma-ceuticals,oral GnRH-antagonists showed promising prospects in the treatment of uterine fibroid.

The gynecological malignancy has a high incidence and mortality. More efficient treatment methods still need to be explored to improve the survival benefits. Artificial intelligence (AI) aims to intelligently process the original problems by simulating the thinking way of human brain. It has obtained significant progress in gynecological malignancy, with great potential in the field of cancer diagnosis and treatment. This paper reviews the application of AI in the diagnosis and treatment of gynecological malignancy, and mainly introduces the research progress on AI in the radiotherapy. This paper mainly focuses on the key issues such as automatic delineation, dose prediction, radiotoxicity prediction and efficacy prediction, and discusses the current benefits and limitations of AI in radiotherapy of gynecological malignancy.

This paper introduced the training system in obstetrics and gynecology(O&G) in the UK and the MRCOG exam organized by the Royal College of Obstetricians and Gynecologists.Comparisons between the O&G specialists training systems of China and UK found that China should better link the resident training and specialists training for a better posteducational medical education system.China should also try to build a China-UK O&G specialist training program to keep pace with the time,for more O&G specialists of international perspectives in China.