Objective To study the expression of hypermethylated gene 1 protein and ovarian cancer gene 1 protein in ovarian cancer and its relationship with the pathological features of ovarian cancer,and to explore its significance in ovarian cancer.Methods Sixty-three cases of ovarian cancer specimens and 63 cases of normal ovarian tissue were taken from January 2014 to December 2015 at the First Hospital of Jilin University.Western blot was used to detect the expression of hypermethylated gene 1 protein and ovarian cancer gene 1 protein in ovarian cancer tissues and normal ovarian tissues,and to analyze the relationship between the expression of two proteins and ovarian cancer.Results The expression of hypermethylated gene 1 protein and ovarian cancer gene 1 protein in ovarian cancer tissues were significantly lower than that in normal ovarian tissues (P＜0.05).There was no significant difference in the expression of hypermethylated gene 1 protein in different staging,different degree of differentiation and different pathological types in ovarian cancer (P＞0.05).The expression of ovarian cancer gene 1 protein in ovarian cancer stage Ⅰ was higher than that in stage Ⅱ and Ⅲ-Ⅳ (P＜0.05).The expression of ovarian cancer gene 1 protein in ovarian cancer stage Ⅱ was higher than that in stage Ⅲ-Ⅳ (P＜0.05).The expression of ovarian cancer gene 1 protein in high differentiation of ovarian cancer was significantly higher than that in moderately differentiated and poorly differentiated (P＜0.05).There was no significant difference between ovarian cancer gene 1 protein expression and poorly differentiated ovarian cancer (P＞0.05).There was no sig nificant difference in the expression of ovarian cancer gene 1 protein in different pathological types of ovarian cancer (P＞0.05).Conclusion Hypermethylatel gene,protein participate in the occurrence of ovarian cancer,the ovarian cancer gene 1 protein is only related to ovarian cancer clinical stage and degree of differentiation.

Objective To isolate side population (SP) cells from an established ovarian cancer (OC)cell line,characterize these cells,and examine their drug resistance. Methods SP and non-SP (NSP) cells were isolated by fluorescence-activated cell sorting (FACS),and cultured in differential conditions,then detected their SP ratio to compare their capability of differentiation and self-renewal. Moreover,SP and NSP cell tumorigenesis was examined by subcutaneous and intraperitoneal injection of these cells to nonobes ediabetic(NOD)-severe combined immundeficient(SCID)mice. Drug resistance to cisplatin was examined by cell counting kit-8 (CCK-8).Results SP cells could be isolated stablly and insistently. There was(4.81 ± 0.43)%of SP cells in the established OC cell line and(4.89 ± 0.33)%of SP cells after cultured the isolated SP cells in differentiation condition,and there was no significant different between these two quantities (P>0.05). However,after cultured the NSP cells,there was only (0.10 ± 0.03)%of SP cells which was significantly lower than that contained in the OC cell line(P<0.01). In the tumorigenesis assay 1.0 × 103 SP cells were injected subcutaneously and formed the xenografted tumors in 6 weeks(3/3),and 1.0×104 NSP cells were injected subcutaneously and did not form xenografted tumors in 12 weeks(0). The tumorigenic capability of SP cells was higher than that of NSP cells(P<0.01). Both the original and the xenografted tumors were low differentiated serous cystadenocarcinomas and expressed the ovarian serous cystadenocarcinomas CA125 marker after stained by HE and immunohistochemistry. Simultaneously,the SP cells were also capable to form tumors as shown by intraperitoneal injection. In the drug resistance assay shown that the 50% inhibitory concentration (IC50) of the SP and NSP cells were respectively(2.33 ± 0.14)μg/ml and(1.60 ± 0.04)μg/ml(P<0.05). After treated the unsorted OC cells with cisplatin,the quantity of SP cells increased to(40.10 ± 4.22)%and there was significant difference,when compared to the untreated cells which was(4.81±0.43)%(P<0.01). The SP cells survival rate was(58.7± 3.3)%when treated with cisplatin at its IC50 dose,and the rate decreased to(7.2±1.3)%(P<0.01)when verapamil was present. Conclusions The SP cells could be isolated from the established OC cell line. They had the capacities of self-renewal,differentiation,and tumorigenesis,and the new tumor demonstrated the original tumor′s phenotype. The SP cells also had stem cells′ biological characteristics and is resistant to cisplatin.

Objective To explore the value of placental alpha-microglobulin-1 in vagina liquid to diagnose premature rupture of membranes. Methods A prospective observational study to initial evaluation included both the standard clinical evaluation for rupture of membranes and placental alpha-microglobulin-1 immunoassay. Rupture of membranes was diagnosed if fluid was seen leaking from the cervical os or if two of the following three conditions were present: pooling of fluid, positive nitrazine test, or feming. Rupture of membranes was diagnosed definitively on review of the medical records after delivery. Results Placental alpha-microglobulin-1 immunoassay confirmed rupture of membranes at initial presentation with a sensitivity of 100% (89/89), specificity of 91% (10/11), positive predictive value of 99% (89/90), and negative predictive value of 100% (10/10),false positive rate of 9% (1/11). Placental alpha-nricroglobulin-1 immunoassay was better than the conventional clinical assessment in confirming the diagnosis of rupture ofmembranes (P<0.01). Conclusion Measurement of placental alpha-microglobulin-1 in cervicovaginal secretions is superior to conventional clinical assessment in the diagnosis of rupture of membranes.

Objective To make sure whether Bcrp1 is the marker of cervical cancer stem-like cells or not by studying the characterization of Bcrp1+ HeLa cells.Methods Immunofluorescence stained flow cytometry and electron microacope were used to sort and observe uhrastructures of Bctp1+ and Bcrp1- HeLa cells.Flow cytometry wag used to identify the cycle and the rate of apoptosis with annexin V in two group cells.The expression of proliferating cell nuclear antigen (PCNA)and caspase-3 were tested using western blot methed.Results (1)There were 7.1% Bcrp1+ cells and 92.9% Bcrol- cells in HeLa cells.Bcrp1+ HeLa cells were large in size of nuclear and nucleoli are clear.and there were rich of cytomicrosome and rough endoplasmic reticulum.After sorted and cultured for 24,48,72 hours,the adhesion in Bcrp1+ cells were 72.8%,81.1%,80.4%,respectively.While,they were 3.3%,18.7%,12.6%at each time for Bcrpl- cells(all P<0. 05 ). (2) There are more S phase cells in Bcrp1+ cells than that in Bcrp1- cells (54. 1% vs 21.1%, P <0. 05) ,while the percentage of G0/G1 and G2/M in Bcrp1 - cells were highter than those in Bcrp1 + cells (53.0% vs 44. 4% ,25.9% vs 1.5% ; all P <0. 05 ). The rate of apoptosis in Bcrp1+ cells was lower than that in Bcrp1 - cells (0. 2% vs 5.3%, P < 0. 05 ). ( 3 ) The expression of PCNA in Bcrp1 + cells was higher than that in Bcrp1- cells (3140 vs 2255, P< 0. 05 ), while the expression of caspase-3 of Bcrpl + cells was lower than that in Bcrp1 - cells ( 1970 vs 3551, P < 0. 05 ). Conclusion There are more vigor and ability of proliferation and lower rate of apeptosis in Bcrp1 + HeLa cells than those in Bcrp1 - cells ,which may be some characters of cervical cancer stem cells.

Objective:Nerve fibres distribution in the functional layer of endometrium of women with endometdosis was investigated.Methods:Histological sections of endometrial tissue were prepared from endometrialcurettings and hysterectomies performed on women with endometnosis(n=25)and without endometriosis(n=40).Immunohistochemistry was used to detect nerve fibres by highly specific polyclonal rabbit antibody PGP 9.5.The assessment of nerve fibre density was performed bv Image Pro Plus Discovery.Results:Nerve fibres were identified throughout the functional layers of the endometrium in all endometriosis patients,but not found in the functional layer of the endometrium in women without endometriosis(P<0.01).Conclusions:Nerve fibres detectad in the functional layer in all women with endometriosis may have important implications for understanding the generation of pain in these patients.

Objective To evaluate endometrial biopsy and curettage in detecting small nerve fibers in eutopic endometrium for diagnosis of endometriosis.Methods Endometrial biopsies and curettings were taken in 65 women with menorrhalgia.Endometrial nerve fibers were immunohistochemically detected using the pan-neuronal marker PGP 9.5.All patients underwent laparoseopie approach.Results The specificity,sensitivity,positive and negative predictive value were 100% of endometrial biopsy and curettings for diagnosis of endometriosis.Condusions Careful endometrial biopsy combined with immunohistochemical staining for nerve fibers may be a reliable means of diagnosing or excluding endometriosis.