Pazopanib Maintenance Therapy in East Asian Women With Advanced Epithelial Ovarian Cancer: Results From AGO-OVAR16 and an East Asian Study.

OBJECTIVE: The recent phase 3 trial AGO-OVAR16 demonstrated that pazopanib maintenance improved median progression-free survival in patients with ovarian cancer whose disease did not progress during first-line treatment. However, this improvement was not seen in the subset of East Asian patients. The current analysis evaluated the efficacy and safety of pazopanib maintenance in East Asian patients from AGO-OVAR16 and a separate East Asian study. MATERIALS AND METHODS: East Asian patients from AGO-OVAR16 (n = 209) and the East Asian study (N = 145) were randomized 1:1 to receive pazopanib 800 mg/d or placebo for up to 24 months. The primary end point for each study was progression-free survival by RECIST (Response Evaluation Criteria in Solid Tumors) based on investigator assessment. Clinical and genetics data were analyzed separately by study or pooled according to separate predetermined statistical plans. RESULTS: Pazopanib maintenance had a detrimental effect on median progression-free survival versus placebo in East Asian patients from the combined studies (n = 354; 17.9 vs 21.5 months; hazard ratio, 1.114; 95% confidence interval, 0.818-1.518; P = 0.4928). Pazopanib maintenance showed a disadvantage in overall survival in East Asian patients from AGO-OVAR16 versus placebo (hazard ratio, 1.706; 95% confidence interval, 1.010-2.883; P = 0.0465); overall survival analysis was not performed in the East Asian study because of insufficient event numbers. Pazopanib-treated patients had a significantly higher incidence of grade 3 or higher hypertension (27%) and neutropenia (13%) versus placebo. CONCLUSIONS: The treatment effect of maintenance pazopanib in East Asian patients seemed to differ from that in non-Asian patients. In study-specific and pooled analyses, none of the potential factors analyzed could satisfactorily explain the different efficacy results of pazopanib in East Asian patients.

[Multicenter randomized controlled clinical study for the operative treatment of malignant ovarian germ cell tumors].

OBJECTIVE: To investigate the operative treatment for first-treated patients with malignant ovarian germ cell tumors who need preservation of fertility. METHODS: The clinical data of 105 patients who were treated with fertility-sparing surgery in 11 hospitals from 1992 to 2010 were collected to evaluate the outcomes of different primary surgical operative procedures. All 105 cases were performed the surgeries that preserved fertility and divided into three groups according to the surgical approaches, comprehensive staging surgery group: 47 cases (44.8%) received comprehensive staging surgeries that including the ipsilateral oophorectomy + omentectomy + retropertoneal lymph node dissection ± appendectomy + multiple biopsies;oophorectomy group:45 cases (42.9%)received ipsilateral oophorectomy ± biopsy of contralateral ovary ± omentectomy;tumor resection group:13 cases (12.4%) received enucleation of the mass with preservation of the ovary. Differences were compared among the three groups of patients in the surgery-related indicators, complications, fertility and prognosis. RESULTS: (1) Surgery-related indicators:the average blood loss of the comprehensive staging surgery group, the oophorectomy group and the tumor resection group were 496, 104 and 253 ml, the mean operation time were 176, 114 and 122 minutes, respectively, and there were significant differences among three groups (P = 0.011, P = 0.000). (2) Complication:the surgical complication rates of the three groups were 17% (8/47), 0 and 1/13, with significant differences (P = 0.015). (3) Reproductive function status: the pregnancy rate and birth rate of the three groups were no significant differences (9/19 vs. 7/19 vs. 2/3, P = 0.515; 8/19 vs. 5/19 vs. 2/3, P = 0.636). (4) PROGNOSIS: the recurrence rate of the three groups were significant differences [13% (6/47) vs. 0 vs. 2/13, P = 0.013], but the death rate with no significant differences [6% (3/47) vs. 0 vs. 0, P = 0.129]; The five-year survival rate of three different groups were 89%, 100% and 100% (P > 0.05), while disease free survival rate were 85%, 100% and 83% (P < 0.05), respectively. CONCLUSIONS: Compared with comprehensive staging surgery, oophorectomy group have higher surgical security and satisfactory prognosis, considerable pregnancy rates and birth rate. The tumor resection security may be reliable, but the prognosis is poor.

Platelet RNA enables accurate detection of ovarian cancer: an intercontinental, biomarker identification study.

Platelets are reprogrammed by cancer via a process called education, which favors cancer development. The transcriptional profile of tumor-educated platelets (TEPs) is skewed and therefore practicable for cancer detection. This intercontinental, hospital-based, diagnostic study included 761 treatment-naïve inpatients with histologically confirmed adnexal masses and 167 healthy controls from nine medical centers (China, n = 3; Netherlands, n = 5; Poland, n = 1) between September 2016 and May 2019. The main outcomes were the performance of TEPs and their combination with CA125 in two Chinese (VC1 and VC2) and the European (VC3) validation cohorts collectively and independently. Exploratory outcome was the value of TEPs in public pan-cancer platelet transcriptome datasets. The AUCs for TEPs in the combined validation cohort, VC1, VC2, and VC3 were 0.918 (95% CI 0.889-0.948), 0.923 (0.855-0.990), 0.918 (0.872-0.963), and 0.887 (0.813-0.960), respectively. Combination of TEPs and CA125 demonstrated an AUC of 0.922 (0.889-0.955) in the combined validation cohort; 0.955 (0.912-0.997) in VC1; 0.939 (0.901-0.977) in VC2; 0.917 (0.824-1.000) in VC3. For subgroup analysis, TEPs exhibited an AUC of 0.858, 0.859, and 0.920 to detect early-stage, borderline, non-epithelial diseases and 0.899 to discriminate ovarian cancer from endometriosis. TEPs had robustness, compatibility, and universality for preoperative diagnosis of ovarian cancer since it withstood validations in populations of different ethnicities, heterogeneous histological subtypes, and early-stage ovarian cancer. However, these observations warrant prospective validations in a larger population before clinical utilities.

[Analysis of the status of DACH1 gene promoter methylation in endometrial carcinoma and its clinical significance].

OBJECTIVE: To analyze the status of DACH1 gene promoter methylation and explore its association with the expression of DACH1 gene promoter methylation and clinical significance of endometrium carcinoma (EC). METHODS: From February 2004 to August 2008, a total of 80 EC tissue samples with comprehensive surgical pathology staging were collected and used for this study. Twenty normal endometrium tissues in 2008 were abstained from the fractional curettage because of dysfunctional uterine bleeding as control. All samples were confirmed pathologically. Methylation specific PCR (MSP) was performed to detect the promoter methylation of DACH1 gene, and analyze its influence on the expression of DACH1 and the relationship between DACH1 promoter methylation and clinicopathological factors in EC. DACH1 protein expression was detected by western blot. Chi-square test and Pearson test were used for statistical analysis. RESULTS: The rate of promoter methylation of DACH1 gene in the EC tissues was significantly higher than that in the normal endometrium issues (30% vs. 5%, P < 0.05). There was an association between the expression of DACH1 and DACH1 gene promoter methylation (r = -0.30, P < 0.01). There was statistical difference between the methylation of DACH1 and the pathological grade (P < 0.05) or histological type (P < 0.05). But DACH1 gene methylation was not related with the age, stage, myometrial invasion depth and lymphnode metastasis (P > 0.05). CONCLUSIONS: DACH1 gene promoter methylaion could lead to a decrease or absence in the DACH1 expression in EC. The promoter methylation of DACH1 gene may induce the inhibition of DACH1 expression, which might be one of the mechanisms of DACH1 gene inactivation in human EC.

[Origin of ovarian epithelial inclusions and its relationship with the development of low-grade serous carcinoma].

OBJECTIVE: To investigate the possible origin of ovarian epithelial inclusions and its relationship with the low-grade ovarian serous carcinoma. METHODS: By comparatively evaluating the morphologic (secretory and ciliated cell distribution) and immunophenotypic [using paired box gene 8 (PAX8), tubulin, calretinin, and Ki-67 as first antibodies] attributes of ovarian epithelial inclusions, the normal tubal epithelium, and the ovarian tumors, all adnexal tissues from a total of 198 patients were studied, including 116 adnexae removed for non-neoplastic indications, 53 serous cystadenomas, 44 serous borderline tumors, and 41 low-grade serous carcinomas, which were collected from Qilu Hospital of Shandong University and University of Arizona in USA. Immunohistochemical single staining was used to detect the expressions of PAX8, tubulin, calretinin, and Ki-67 in the two groups, while immunohistochemical double staining of PAX8/calretinin was used to figure out the immunophenotype of various ovarian epithelial inclusions in a more intuitive way. RESULTS: With immunohistochemical single staining of PAX8 and calretinin, the vast majority (90%, 54/60) of ovarian surface epithelia displayed a mesothelial phenotype [calretinin(+), PAX8(-)], whereas 10% (6/60) of the cases displayed foci with tubal phenotype [calretinin(-), PAX8(+)]. In contrast, most (79%, 728/921) of the ovarian epithelial inclusions displayed a tubal phenotype, though 21% (193/921) of the ovarian epithelial inclusions showed a mesothelial phenotype. It was further proved by immunohistochemical double staining of PAX8/calretinin. Secretory and ciliated cells were found in the ovarian epithelial inclusions with tubal phenotype. There was a progressive increase in the secretory/ciliated cells ratio and proliferative index, from ovarian epithelial inclusions/cystadenomas to borderline tumors to low-grade serous carcinoma, according to the expression of tubulin and Ki-67. CONCLUSIONS: The findings make a strong argument that the ovarian epithelial inclusions displaying a tubal phenotype with PAX8(+), calretinin(-) is likely derived from fallopian tube rather than through Mullerian metaplasia from ovarian surface epithelium. The increasing trend of secretory/ciliated cells ratio and proliferative index from ovarian epithelial inclusions/cystadenomas to borderline tumors to low-grade serous carcinoma indicates that the latter is a clonal expansion of secretory cells. Genetic and molecular studies are needed to further confirm these findings.

[Application of pathological diagnosis by rapid paraffin sections for biopsy in the diagnosis and treatment of cervical diseases].

OBJECTIVE: To evaluate the application of pathological diagnosis by rapid paraffin sections in the diagnosis and treatment of cervical diseases. METHODS: A total of 176 cases from our hospital between September 2009 and January 2010 with abnormal cervical cancer screening (including abnormal cytology result and high-risk HPV continuous positive) were randomly divided into 2 groups. Eighty-seven cases of them whose biopsy were got by Belinson forceps under the direction of colposcopy with rapid paraffin sections by ultrasonic histopathological rapid processor and BT transparent agents were selected as group A, while 89 cases with conventional paraffin sections were selected as group B. The production time and quality for paraffin sections were analyzed in the two groups. Those diagnosed as cervical intraepithelial neoplasia (CIN) II or even worse and some special patients with CINI in the two groups received surgery, including loop electrosurgical procedure (LEEP), cold knife conization (CKC), hysterectomy or radical hysterectomy. Tissue obtained after surgery was sent for routine pathological examination. If the results of postoperative routine pathological examination were inconsistent with the rapid or routine biopsy pathological examination, the heavier results were regard as the final diagnoses. The pathological results and diagnose accordance rates were recorded and compared between group A and group B. RESULTS: The quality of sections in two groups were all satisfied or basically satisfied to meet the diagnostic requirements. There were statistically significant difference in average production time between group A and B (40 minutes vs 24 hours, P<0.05). Thirty patients in group A and 32 patients in group B received surgery. The coincidence rate of biopsy pathological results and final diagnoses were 93% (28/30) for group A and 91% (29/32) for group B, in which there were not statistically significant difference (P>0.05). CONCLUSION: Rapid paraffin sections technology is safe, accurate and economical for rapid pathological diagnosis of cervical diseases, which is worthy for being widely used in hospitals.

Expression of BAFF in the trophoblast and decidua of normal early pregnant women and patients with recurrent spontaneous miscarriage.

BACKGROUND: BAFF, the B cell activation factor, is a member of the tumor necrosis factor (TNF) ligand family that binds to BCMA, TACI, and BAFF-R. Previous studies have shown that members of the TNF family are detected in human placental trophoblast cells, but the expression patterns of BAFF involved in human decidua and the differential expression of BAFF between normal pregnancy and miscarriage are still incompletely documented or unknown. This study was designed to investigate the expression of BAFF and BAFF-R in the trophoblast and decidua of normal early pregnant women and recurrent spontaneous abortion (RSA) patients. METHODS: Forty-five patients with RSA and 45 normal pregnant women were included in this study. By reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemical experiments, we explored the expression of BAFF and BAFF-R in the maternal-fetal interface of normal early pregnant women and RSA patients. RESULTS: Analysis by RT-PCR and Western blotting revealed that BAFF was detected in both trophoblast and decidua of all the samples, and the expression level was higher in the tissues of normal early pregnant women (P<0.05) than that of recurrent spontaneous abortion patients under the same gestational weeks. Messages for BAFF-R were absent. Immunohistochemical experiments showed that expression of BAFF was cell-specific which was localized to villous cytotrophoblast and syncytiotrophoblast cells in trophoblast and to stromal cells in decidua. Whereas BAFF was prominent on the trophoblast and decidua of normal early pregnant women, it was decreased in the tissues of RSA patients. CONCLUSIONS: BAFF might steer maternal leukocytes away from a harmful immune response and toward a favorable one and play a potentially vital role for successful pregnancy.

[Expression and significance of chemokine CXC receptor 3, 4 and their ligands at the early pregnancy decidua and villi].

OBJECTIVE: To explore the expression and significance of chemokine CXC receptor (CXCR) 3 and CXCR4 and their ligands (CXCL) at the early pregnancy decidua and villi. METHODS: Decidual mononuclear cells were isolated from the normal decidua of 5 - 8 weeks pregnant women by lymphocyte separation medium in vitro. CD(56)(+) natural killer (NK) cells were purified by dynabeads cell sorter kit. Purity and phenotype of CD(56)(+) decidua NK cells were analyzed by fluorescence-activated cell sorter (FACS). Gene expression of CXCR3 and CXCR4 in decidua NK cells and CXCL9, CXCL10 and CXCL12 in early pregnancy decidua and villi was assessed by RT-PCR. Protein expression of CXCL9, CXCL10 in normal endometrium and early pregnancy decidua was characterized and quantified by streptavidin-biotin peroxidase chain reaction (SP) immunohistochemistry and computered image analysis system. Correlations between the gray degree of CXCL9 and CXCL10 and the number of CD(56)(+)NK cells in upper tissue were analyzed by Spearman's correlation coefficient rank test. RESULTS: The phenotype of 98.7% decidua NK cells was CD(56)(bright). The genes of CXCR3 and CXCR4 were expressed in decidua NK cells and that of CXCL9 and CXCL10 were expressed in early pregnancy decidua and CXCL12 in early pregnancy villi. CXCL9 and CXCL10 were expressed in the cytoplasm of surface epithelia, glandular epithelia and stromal cells of early pregnancy decidua and were not expressed in villi by immunohistochemistry. The gray degree of CXCL9 and CXCL10 in the secretory phase endometrium (56 +/- 43, 59 +/- 47) was stronger than that in the proliferative phase (16 +/- 18, 8 +/- 14, P < 0.05) and reached the highest (143 +/- 35, 158 +/- 29, P < 0.05) in the early pregnancy decidua. The number of CD(56)(+) NK cell in the secretory phase endometrium (60 +/- 20) was more than that in the proliferative phase endometrium (23 +/- 4, P < 0.05) and was the most in the early pregnancy decidua (114 +/- 15, P < 0.05). The gray degree of CXCL9 in upper tissue had a positive correlation with the number of CD(56)(+) cells (r = 0.88, P < 0.05) and that of CXCL10 had a similar pattern to CXCL9 (r = 0.86, P < 0.05). CONCLUSION: The interactions between CXCL9, CXCL10 and CXCL12 expressed in decidua and villi and CXCR3, CXCR4 expressed in CD(56)(+) decidua NK cells may influence the CD(56)(+)NK cell recruitment at the maternal-fetal interface.

[Effect of extracellular signal regulated kinase signal pathway on apoptosis induced by MG262 in ovarian cancer cells].

OBJECTIVE: To investigate whether the proteasomes inhibitor MG262 exerts its anti-cancer function by inducing apoptosis in human ovarian cancer cells, and whether the extracellular signal regulated kinase (ERK) signaling pathway is involved in the regulation of apoptosis induction. METHOD: Human ovarian cancer cell line SKOV3 was incubated with different concentrations of MG262 for 24 and 48 hours. Cell viability was evaluated with 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay at different time points of culturing. Flow cytometry was used to detect cell apoptosis rate. The expression of vascular endothelial growth factor (VEGF) was evaluated with western blot and enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the expression of phosphorylated ERK (p-ERK). RESULTS: The viability of SKOV3 cells was decreased by MG262 in a concentration-dependent fashion (P < 0.05). After 24 h incubation with MG262 at 1, 10, 20, 40, 60 and 80 nmol/L, the viability rates of SKOV3 were (94.6 +/- 3.1)%, (92.7 +/- 3.7)%, (89.5 +/- 7.7)%, (84.2 +/- 5.1)%, (82.0 +/- 7.4)% and (76.8 +/- 11.0)% respectively, and after 48 h incubation, those figures were further decreased to (91.3 +/- 10.1)%, (86.8 +/- 4.5)%, (74.6 +/- 4.2)%, (56.8 +/- 2.1)%, (49.3 +/- 4.5)% and (37.4 +/- 5.4)%, respectively (P < 0.05). Apoptosis rate of SKOV3 cells induced by MG262, PD98059 or their combination was (30.7 +/- 4.3)%, (26.8 +/- 8.6)% and (50.3 +/- 10.6)%, respectively, which were significantly different compared with controls (P < 0.05). In contrast to SKOV3 cells, apoptosis rate of 293T cells induced by MG262, PD98059 or their combination was (14.5 +/- 5.3)%, (16.2 +/- 7.5)% and (10.8 +/- 7.3)%, respectively, which were not significantly different compared with controls (P > 0.05). p-ERK expression decreased gradually in a time-dependent manner. And wild-type p53 expression was not significantly different. There was no significant difference between experimental and control 293T cells (P < 0.05). In addition, MG262 down-regulated VEGF secretion and expression in SKOV3 cells (P < 0.05). CONCLUSIONS: Proteasome inhibitors can induce apoptosis and inhibit cell proliferation and angiogenesis through ERK signal pathway in SKOV3 cells.

[Treatment of intraperitoneal implanted human ovarian carcinoma of nude mice by angiostatin gene and chemotherapy in vivo].

OBJECTIVE: To observe the effects of angiostatin gene combined with chemotherapy on implanted human ovarian carcinoma of nude mouse. METHODS: The mice were randomly divided into four groups after 7 days of the intraperitoneal injection of tumor cells (4 x 10(6)), and injected respectively with empty plasmid pcDNA3.0, angiostatin plasmid, cisplatin, and angiostatin plasmid + cisplatin. For combinational treatment, reagents were delivered in a timed fashion, where angiostatin plasmid was injected first, followed by cisplatin 24h later. The tumor samples were prepared to be used in the examinations of the expression of angiostatin with immunohistochemistry, of MVD in the tumor with immunohistochemistry, and of cell apoptosis with TUNEL staining. RESULTS: Tumor growth and ascites formation were inhibited in all 3 groups except for the control group. The therapeutic effectiveness in the combined group was more significant than in the other two groups. In this group, MVD (32.5 +/- 4.3) was the lowest and apoptosis index (5.12 +/- 0.63) was the highest (P < 0.01). CONCLUSIONS: Angiostatin gene therapy combined with chemotherapy has a synergistic effect on the inhibition of ovarian cancer angiogenesis and ascites formation. Combining multiple therapies to treat ovarian cancer is an effective strategy.

Development and evaluation of novel tumor-targeting paclitaxel-loaded nano-carriers for ovarian cancer treatment: in vitro and in vivo.

BACKGROUND: Ovarian cancer is the most leading cause of death and the third most common gynecologic malignancy in women. Traditional chemotherapy has inevitable drawbacks of nonspecific tumor targeting, high toxicity, and poor therapeutic efficiency. In order to overcome such shortcomings, we prepared a novel nano-carrier drug-delivery system to enhance the anti-tumor efficiency. METHODS: In vitro characterizations of nano-carriers were determined by TEM, DLS. Cell viability was measured by MTT method. RT-PCR was performed to measure the expression of FARα in three ovarian cancer cell lines. The drug-release study and the uptaken study were measured in vitro. The pharmacokinetic and the drug distribution study were verified by HPLC methods in vivo. The enhanced anti-tumor efficiency of FA-NP was evaluated by the tumor inhibitory rate in vivo. RESULTS: Paclitaxel (PTX)-loaded nanoparticles (NPs) (PTX-PEG-PLA-NP and PTX-PEG-PLA-FA-NP) were prepared successfully, and the drug-release study showed that the cumulative release rates of NP groups were much less than free PTX group. The pharmacokinetic study showed that the elimination phase of two kinds of NP groups were much longer than that of PTX group. The drug distribution in different tissues showed that the peak-reach time was 2 h in the PTX group and 6 h in both NP groups. All of these results confirmed the excellent slow-release effects of both kinds of nano-carriers. More importantly, we confirmed that PTX-PEG-PLA-FA-NP had greater uptake by SK-OV-3 cells than PTX-PEG-PLA-NP and free PTX in vitro. A drug-distribution study of tumor-bearing mice demonstrated that the PTX concentration of tumor tissues in the PTX-PEG-PLA-FA-NP group was 3 times higher than the other two groups. PTX-PEG-PLA-FA-NP was uptaken much more by SK-OV-3 cells than PTX-PEG-PLA-NP and free PTX. Eventually, based on the slow-release effect and tumor-targeting characteristics of PTX-PEG-PLA-FA-NP, a cytotoxicity test indicated that PTX-PEG-PLA-FA-NP was much more toxic to SK-OV-3 cells than the controls. The tumor inhibitory rate in the PTX-PEG-PLA-FA-NP group of tumor-bearing mice was about 1.5 times higher than the controls. The tumor targeting and anti-tumor efficiency of PTX-PEG-PLA-FA-NP were confirmed both in vitro and in vivo. CONCLUSIONS: We developed an ovarian cancer targeting nano-carrier drug delivery system successfully, which showed perfect ovarian cancer targeting and anti-tumor effect, thus have the potential to be a new therapy strategy for ovarian cancer patients.

[Inhibitory effect of breast cancer metastasis suppressor l gene on metastasis of human ovarian cancer cell in vitro and in vivo].

OBJECTIVE: To study the inhibition effects of breast cancer metastasis suppressor l (BRMS1) gene on metastasis of human ovarian cancer cell in vivo and in vitro. METHODS: BRMS1 gene was transfected into human ovarian cancer cell line A2780 by liposome transfection method. The cells were divided into 3 groups: transfected group with pcDNA3-BRMS1, negative control group with empty plasmid pcDNA3, blank control group without any transfection. Proliferation, apoptosis, growth rate, capability of colony formation, gap junctional intercellular communication (GJIC), capability of adhesion, changes of surface and internal structures of cells were observed in vitro. The cells were injected into athymic mice to establish animal tumor models, and inhibition of the tumorigenicity and metastasis were observed in vivo. RESULTS: BRMS1 gene was transfected into A2780 cell line successfully. There were no significant differences in the growth rate among transfected group, negative control group and blank control group (P > 0.05). The amounts of cell clones per well were 40 +/- 4 in transfected group, 42 +/- 7 in blank control group and 39 +/- 4 in negative control group (P > 0.05 between each two groups). The ratios of S vs G(2)/M and G(0)/G(1) were not significantly different in three groups (P > 0.05). The ultramicrostructure of cells detected by electron microscope showed that GJIC function in transfected group was higher than that in the other two groups. While in migration assay, the numbers of cells in lower chamber passing through the membrane in transfected group, blank control group and negative control group were 112 +/- 23, 306 +/- 49 and 322 +/- 91, respectively; with significant differences among 3 groups (P < 0.01). CONCLUSION: BRMS1 gene could suppress metastasis of ovarian cancer in vitro and in vivo.

[Combination chemotherapy with etoposide and cisplatin for high-risk, chemorefractory and recurrent gestational trophoblastic neoplasia].

OBJECTIVE: To evaluate the feasibility and effectiveness of combination chemotherapy with etoposide and cisplatin (EP) regimen on the patients with high-risk, chemorefractory and recurrent gestational trophoblastic neoplasia (GTN). METHODS: Thirty-nine patients with gestational trophoblastic tumors were analyzed retrospectively, 25 of 39 patients were of high-risk, 9 patients were chemorefractory and 5 patients were recurrent. All 39 patients were administrated with EP regimen, and 10 patients were assisted with surgery. All the patients were followed up. Clinical response, toxicity, the occurrence of secondary tumors of all patients, and the fertility of 30 patients whose fertility function was preserved were investigated. RESULTS: Thirty-nine GTN patients underwent a total of 221 cycles of the EP regimen. The average number of courses for each patient was 5.7. The total complete remission rate of the regimen was 74% (29/39). Twenty-five patients with high-risk GTN received a total of 139 cycles and the average number of courses was 5.6. Nineteen patients achieved complete remission and 6 patients showed drug-resistant. The complete remission rate of the high-risk group was 76% (19/25). Nine patients with chemorefractory GTN obtained a total of 55 cycles and the average number of courses was 6.1. Six patients achieved complete remission and 3 patients showed drug-resistant again. The complete remission rate of the chemorefractory group was 6/9. Five patients with recurrent GTN received 27 cycles and the average number of courses was 5.4. Four patients achieved complete remission, 1 patient showed drug-resistance and died. Bone marrow toxicity, gastrointestinal reaction and alopecia were the main side effects of the EP regimen, but the bone marrow toxicity was slight and no grade IV side effect occurred. No fatal effect was found. Eight of 30 patients whose fertility fuction was preserved had become pregnant after recovery, with a total of 8 pregnancies. Among them, 2 were terminated by induced abortion, and 6 underwent normal term delivery and gained 6 infants who had no congenital malformation. All the 6 children had normal growth and development after childbirth. None of the women developed secondary tumors. CONCLUSION: The EP regimen is effective and safe for the treatment of high-risk, chemorefractory and recurrent GTN.

[Clinical study of topotecan and cisplatin as first line chemotherapy in epithelial ovarian cancer].

OBJECTIVE: To evaluate efficacy and toxicity of topotecan and cisplatin (TP) as first line chemotherapy in epithelial ovarian cancer, and its effect on prognosis of the patients. METHODS: Totally 94 eligible patients with pathologically verified stage II - IV epithelial ovarian cancer were enrolled into 3 groups of this clinical trial. (1) TP group: 30 patients were treated with topotecan, 0.75 mg.m(-2).d(-1), for 5 days, and cisplatin, 75 mg/m(2), on day 1. (2) Paclitaxel and carboplatin (TC) group: 31 patients were treated with paclitaxel, 135 mg/m(2), on day 1, and carboplatin, given to an area under the curve (AUC) of 5, on day 1. (3) Cyclophosphamide and cisplatin (PC) group: 33 patients were treated with cyclophosphamide, 500 mg/m(2), on day 1, cisplatin 75 mg/m(2), on day 1. Cycles were repeated every 21 - 28 days. EFFICACY of the three combination regimens were evaluated after 6 - 8 courses. RESULTS: (1) EFFICACY: the overall response rate (ORR) in the TP group was 70%. Of the 30 patients, 8 achieved a complete response (CR) and 13 a partial response (PR). The ORR in the TC group was 77%. Of the 31 patients, 10 achieved a CR and 14 a PR. While the ORR in the PC group was 42%. Of the 33 patients, 5 achieved a CR and 9 a PR. There was no significant difference in clinical efficacy between TP group and TC group (P > 0.05). But there was a significant difference between TP group and PC group (P < 0.05). (2) Disease free survival (DFS): after median follow-up of 25 months, one-year disease free survival rate was 67% in TP group, 71% in TC group and 42% in PC group (P > 0.05). Two-year disease free survival rate was 57% in TP group, 64% in TC group and 39% in PC group (P > 0.05). (3) Overall survival (OS): One-year survival rate was 93% in TP group, 97% in TC group and 91% in PC group (P > 0.05). Two-year survival rate was 77% in TP group, 84% in TC group and 67% in PC group (P > 0.05). (4) TOXICITY: Grade III - IV myelosuppression was 60% (18/30) in TP group, 26% (8/31) in TC group and 30% (10/33) in PC group. The TP regimen had the greatest hematologic toxicity (P < 0.05). Nonhematologic toxicities were not significantly different among the three regimens (P > 0.05). CONCLUSIONS: As first line chemotherapy in epithelial ovarian cancer, TP regimen comparable to the standard chemotherapy regimen.

[A multicenter randomized clinical trial of weekly paclitaxel and carboplatin vs three weeks paclitaxel and carboplatin as first line chemotherapy in epithelial ovarian cancer].

OBJECTIVE: To compare the efficacy of combination regiments of taxol given weekly plus carboplatin and taxol given every three weeks plus carboplatin. To observe the toxicity of the two regiments. To observe the two-year survival rate in the two groups. METHODS: Total 125 eligible patients in 13 centers of CGOG were entered into the two arms of this randomized clinical trial, of whom 51 were entered into weekly taxol group and 74 entered into 3 weeks taxol group. RESULTS: 81.6% (102/125) of patients had satisfactory decreasing of CA125 level after optimal cytoreductive surgery and chemotherapy. 86.3% (44/51) of patients is in weekly group and 78.4% (58/74) of patients in three weeks group (P > 0.05). Relapse frequency is 29.7% in every three weeks group and 19.6% in weekly group (P > 0.05). Median interval to relapse is 15.7 months in every three weeks group and 13.6 months in weekly group (P > 0.05). One-year survival rate is 95.2% in every three weeks and 93.9% in weekly group (P > 0.05). Two-year survival rate is 78.7% in every three weeks and 85.3% in weekly group (P > 0.05). Grade III and IV myelosuooression is 45.9% in three weeks group and, 27.5% in weekly group (P < 0.05). CONCLUSION: (1) The two regiments had equal efficacy. (2) Myelosuppression was less frequency in the weekly group than in every three weeks group. (3) Weekly taxol therapy has mild toxicity and is more suitable for the old and feeble patients. Weekly taxol therapy can be conveniently administered in outpatients department.

[Procaspase-3 enhances in vitro effect of cytosine deaminase-thymidine kinase fusion disuicide gene therapy system on human ovarian carcinoma].

OBJECTIVE: To investigate whether procaspase-3 could enhance the in vitro effect of cytosine deaminase-thymidine kinase/5-fluorocytosine + ganciclovir (CD-TK/5-FC + GCV) system in human ovarian cancer cells. METHODS: Eukaryotic expression vectors containing procaspase-3 gene (pcDNA3-casp3) and CD-TK fusion disuicide genes regulated by human telomerase reverse transcriptase (hTERT) promoter (pBTdel-279-CD-TK) were constructed. Western blot was used to detect the expression of procaspase-3 protein in 3AO cells with transfection of pcDNA3-casp3 (3AO-pcDNA3-casp3) or pcDNA3 (3AO-pcDNA3). RT-PCR was used to detect the expression of CD and TK genes in 3AO-pcDNA3-casp3 or 3AO-pcDNA3 cells after transfection with pBTdel-279-CD-TK or pBTdel-279. Following the treatment with 5-FC + GCV, the cell survival rate and cell cycle distribution were detected by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry (FCM). Fluorogenic substrate assay and western blot were used to detect caspase-3 activity in these cells. RESULTS: Procaspase-3 protein was expressed in 3AO-pcDNA3-casp3 cells, while not in 3AO-pcDNA3. The expressions of CD and TK genes were observed in both 3AO-pcDNA3-casp3 and 3AO-pcDNA3 after transfection with pBTdel-279-CD-TK. After treatment with 5-FC + GCV, the survival rate of 3AO-pcDNA3-casp3 + pBTdel-279-CD-TK was significantly lower than that of 3AO-pcDNA3 + pBTdel-279-CD-TK cells. After treatment with 5-FC (2 mmol/L) + GCV (10 microg/ml), there was a higher apoptotic ratio (37.98%) and S-phase block (49.67%) in 3AO-pcDNA3-casp3 + pBTdel-279-CD-TK than in 3AO-pcDNA3 + pBTdel-279-CD-TK cells (21.34% and 35.76%, respectively) (P < 0.05). Following the action of 5-FC (1 mmol/L) + GCV (1 microg/ml) for 48 h, the relative activity of caspase-3 in 3AO-pcDNA3-casp3 + pBTdel-279-CD-TK cells was 189.7, significantly higher than 44.9 in 3AO-pcDNA3 + pBTdel-279-CD-TK cells (P < 0.05). CONCLUSION: Co-expression of procaspase-3 may lead to a significant enhancement of the efficacy of CD-TK fusion gene therapy.

Effect of human umbilical cord mesenchymal stem cells transplantation on nerve fibers of a rat model of endometriosis.

BACKGROUND: Endometriosis is a common, benign, oestrogen-dependent, chronic gynaecological disorder associated with pelvic pain and infertility. Some researchers have identified nerve fibers in endometriotic lesions in women with endometriosis. Mesenchymal stem cells (MSCs) have attracted interest for their possible use for both cell and gene therapies because of their capacity for self-renewal and multipotentiality of differentiation. We investigated how human umbilical cord-MSCs (hUC-MSCs) could affect nerve fibers density in endometriosis. MATERIALS AND METHODS: In this experimental study, hUC-MSCs were isolated from fresh human umbilical cord, characterized by flow cytometry, and then transplanted into surgically induced endometriosis in a rat model. Ectopic endometrial implants were collected four weeks later. The specimens were sectioned and stained immunohistochemically with antibodies against neurofilament (NF), nerve growth factor (NGF), NGF receptor p75 (NGFRp75), tyrosine kinase receptor-A (Trk-A), calcitonin gene-related peptide (CGRP) and substance P (SP) to compare the presence of different types of nerve fibers between the treatment group with the transplantation of hUC-MSCs and the control group without the transplantation of hUC-MSCs. RESULTS: There were significantly less nerve fibers stained with specific markers we used in the treatment group than in the control group (p<0.05). CONCLUSION: MSC from human umbilical cord reduced nerve fiber density in the treatment group with the transplantation of hUC-MSCs.