Impact of Conditioned Medium of Decidual Stromal Cell Cluture on Proliferation,Apoptosis and Invasion of Trophoblast Cells / 实用妇产科杂志

Objective:To investigate the impact of conditioned medium of decidual stromal cell culture(DSCM) on proliferation,apoptosis and invasion of trophoblast cells.Methods:lsolution,culturing and indentifying the purity of decidual stromal cells from healthy women of early pregnancy to prepare different concentrations of DSCM (5％,20％,50％ and 100％);Proliferation was evaluated by MTT assay;Cell apoptosis was examined by flow cytometry using annexin V-FITC/PI staining;Cell invasion was assayed using a Transwell chamber;RT-PCR and gelatin zymography were used to explore the mechanism of cell invasion.Results:Using this method we obtained a purity over 90％ of decidual stromal cells.With the concentration of DSCM increased(5％,20％,50％,100％),the OD value of the MTT assay has elevated and the rate of apoptosis declined,the infiltrating cell increased as well (P ＜ 0.05);Compared with control group,DSCM treatment led to a significant increase in the mRNA and protein expression of MMP-2/9(P ＜0.05).Conclusions:Decidual microenviroment may promote trophoblast proliferation and inhibite its apoptosis besides could enhance its ability of invasion by altering the expression of MMP-2 and MMP-9.

Expression and significance of miR-155 and CXCR4 in placenta tissues from patients with preeclampsia / 国际检验医学杂志

Objective To investigate the expression changes and clinical significance of microRNA-155(miR-155) and chemokine receptor 4(CXCR4) in placental tissue from the patients with preeclampsia(PE).Methods Thirty pregnant women with severe PE(sPE) in the Third Affiliated Hospital of Zhengzhou University from December 2015 to February 2016 served as the sPE group,and contemporaneous 30 healthy pregnant women undergoing cesarean section due to the social factors served as the healthy control group(N).The real-time fluorescent quantitative PCR(RT-PCR) was used to detect the expression of miR-155 and CXCR4 mRNA in placental tissue and the relationship between miR-155 and CXCR4 levels was analyzed.The immunohistochemistry SABC methods were used to detect the expression of CXCR4 protein in villous cytotrophoblast(VCT) tissue microarray(TMA,42 cases in the normal control group 1,56 cases in the PE group) and extravillous cytotrophoblast(EVCT) TMA(29 cases in the normal control group 2,47 cases in the PE group) constructed by the same research group.Results (1) There was no statistically significant difference in the age,gestational age and pre-pregnancy body mass index(BMI) between the two groups(P>0.05).The differences of blood pressure between the two groups were statistically significant(P0.05).The gestational weeks of the PE group were earlier than those in the N group 1,the systolic/diastolic blood pressure and urine protein were higher than those in the N group,the differences all were statistically significant(P<0.05),the neonatal birthweight was significantly lower than that in the N group with statistically significant differences(P<0.05).(3)The expression level of miR-155 mRNA in placental tissue in the sPE group was 1.53±0.92,which was significantly higher than 0.87±0.73 in the control group,the difference between the two groups was statistically significant(P<0.05).(4) The expression level of CXCR4 mRNA in the N group was 1.51±1.85,which in the sPE group was 0.54±0.38,the difference between the two groups was statistically significant(P<0.05).(5) In the sPE group,the miR-155 level and CXCR4 level in placntal tissue had a significant correlation(r=-0.773,P<0.05).(6) CXCR4 protein was expressed in VCT and EVCT TMA;the CXCR4 positive expression rate of the PE group in VCT TMA was 48.21%(27/56),which in the sPE group was 47.92%(23/48) and which in the early onset PE group was 53.66%(22/41),which all were significantly lower than 83.33%(35/42)in the normal control group,the differences all were statistically significant(P<0.05).(7)The positive expression rate of CXCR4 in the PE group in placent EVCT TMA was 48.94%(23/47),which in the sPE group was 50.00%(22/44) and which in the early PE onset group was 52.63%(20/38),which all significantly lower than 79.31%(23/29) in the normal control group,the differences all were statistically significant(P<0.05).Conclusion The level of placental tissue miR-155 is increased in the patients with sPE,while the level of CXCR4 is decreases obviously,both have a negative correlation,which may be associated with the pathogenesis of PE.

Effect of SHBG gene on the apoptosis of human trophoblastic cells / 局解手术学杂志

Objective To investigate the effects of sex hormone-binding globulin (SHBG)gene in the apoptosis of human trophoblastic cells.Methods The siRNA specific-targeting SHBG gene was transfected into human trophoblastic cells and they were divided into six groups:trophoblasts without transfection in normal control groups(group Ⅰ);transfect liposome in blank control groups(group Ⅱ);transfect nonspecific siRNA in negative control groups(group Ⅲ);transfect SHBG siRNA-Ⅰ,SHBG siRNA-Ⅱ,SHBG siRNA-Ⅲ respectively in trans-fection group(group Ⅳ,Ⅴ,Ⅵ).Hoechst 33258 dying method was used to detect cell apoptosis.SHBG and Caspase-3 mRNA profiling and the level of SHBG and caspase-3 protein were detected by real-time PCR and Western blot.Results There was no statistical significant difference in the gene expression and protein level of SHBG and caspase-3 in group Ⅰ,Ⅱ and Ⅲ (P >0.05).In Ⅳ,Ⅴ and Ⅵ group,there was no statistical significant difference in the expression level of SHBG and caspase 3 (P >0.05).Compared with group Ⅰ,Ⅱ and Ⅲ,the a-mount of SHBG gene expression decreased obviously,the caspase-3 mRNA and protein level increased obviously and the trophoblast cell ap-optosis increased markedly (P <0.05).Conclusion Through siRNA interference technology can reduce SHBG gene expression in human trophoblastic cells,and it can lead to excessive apoptosis of human trophoblasts cells.

Epithelioid trophoblastic tumor: a case report of a rare trophoblastic neoplasm.

Epithelioid Trophoblastic Tumor (ETT) is a rare neoplasm of the chorionic type intermediate trophoblastic cells. It is a neoplasm of reproductive age women and usually follows a gestational event. ETT can occur at both intra uterine and extra uterine sites and can be confused with other entities such as squamous cell carcinoma, placental site nodule, placental site trophoblastic tumor etc. Hence, proper diagnosis of this tumor is necessary to avoid unnecessary, excessive treatment as surgical treatment is considered sufficient for ETT. We present a case of ETT in a 36 year old female, who came with symptoms of pain abdomen, white discharge per vaginum and a cervical mass.

Effect of Ephrin-A1/EphA2 on Invasion of Trophoblastic Cells / 华中科技大学学报（医学）（英德文版）

The effect of axon guidance factors ephrin-A 1/EphA2 on the invasion of trophoblastic cells and the possible mechanism were investigated in this study.The expression of EphA2 in vascular endothelial cells was detected by immunohistochemistry.The proliferation and invasion of TEV-1 cells (an extravillous trophoblastic cell line) in first trimester were determined by cell counting kit-8 (CCK-8)and Transwell invasion assay.Real-time PCR was used to detect the expression of ephrin-A1 in TEV-1cells treated with EphA2 at different concentrations (10,50,100,500,1000 and 5000 μg/L).The results showed:(1) EphA2 was expressed in the vascular endothelial cells; (2) EphA2 could promote the proliferation of TEV-1 cells.The proliferative capacity reached a peak in TEV-1 cells treated with 100 μg/L EphA2 (P＜0.05); (3) EphA2 could increase the invasion of TEV-1 cells.The invasive ability was the greatest in TEV-1 cells treated with 500 μg/L EphA2 (P＜0.05); (4) in the presence of EphA2 (0-500μg/L),the expression of ephrin-A1 was increased concentration-dependently (P＜0.05),but when the concentration of EphA2 was over 500 μg/L,the expression of ephrin-A 1 ceased to increase (P＞0.05).It was concluded that EphA2 can promote the invasion and proliferation of the human extravillous trophoblastic cells probably by regulating the ephrin-A1 ligand.

Effect of Toxoplasma gondii ME49 strain on apoptosis of mouse placental trophoblastic cells / 第二军医大学学报

Objective: To investigate the effect of Toxoplasma gondii ME49 strain on the apoptosis of mouse placental trophoblastic cells in vitro. Methods: Mouse placental trophoblastic cells (concentration of 5×106/ml) were cultured in the different cell culture vessels. The cells were treated for 8 h with different concentrations of Toxoplasma gondii ME49 strain (the concentration of tachyzoites was 2×106/ml, 4×10 6/ml, and 8×106/ml, respectively). FCM was used to examine the apoptosis rates of the placental trophoblastic cells stained with the fluorescent dye of Annexin V-FITC/PI; fluorescence microscopy was used to observe the changes of cellular morphology, and Western blotting analysis was used to detect the protein levels of Bax and Bcl-2. Results: The trophoblastic cells infected with Toxoplasma gondii ME49 strain showed a higher apoptosis compared to the normal cells(P<0.05), and the apoptosis rates increased with the concentration of tachyzoites in the infected groups. The highest apoptosis rate was 28.37% which was found 8 h after culture with 8×106/ml tachyzoites. Fluorescence microscope observed that the apoptosis of trophoblastic cells increased with the increase of Toxoplasma gondii. Western blotting analysis showed that the relative expression levels of Bax and Bcl-2 were 1.24±0.05, 1.37±0.03, 1.78±0.04, and 1.15±0.03, 1.09±0.05, 0.97±0.01, respectively, which were significantly different from those of the control group (1.17±0.06, 1.23±0.02, P<0.05). Conclusion: Infection with Toxoplasma gondii ME49 strain can promote the apoptosis of mouse placental trophoblastic cells in vitro through up-regulating Bax expression and down-regulating Bcl-2 expression.

Study on retinol binding protein (RBP) receptor in hydatidiform mole trophoblastic cells.

Hydatidiform mole is an abnormal pregnancy characterized by the proliferation of cytotrophoblastic, syncytiotrophoblastic, and intermediate trophoblastic cells in histological specimens. Vitamin A plays a role in controlling cell proliferation, and decrease in vitamin A level will cause an uncontrollable proliferation. To date, it is not known whether there is a relationship between vitamin A deficiency and hydatidiform mole. This study aimed to demonstrate the presence of retinol binding protein (RBP) receptors in the hydatidiform mole trophoblastic cells, that would provide explanation on the relationship of vitamin A and hydatidiform mole. The study was a descriptive study. The specimens of the study were paraffin blocks of hydatidiform mole made in 2005, and the examinations were performed by indirect immunohistochemistry. We examined the distribution of the cells showing expression of RBP receptor, the strength of expression, and location of the expression. As many as 21 specimens were collected, and the distributions of RBP receptor expression in hydatidiform mole trophoblastic cells ranged from moderate to dense. The expression in syncytiotrophoblastic cells was stronger than that in cytotrophoblastic cells. Furthermore, the expressions were found in the cell membranes and cytoplasm.

A case of placental site trophoblastic tumor / 대한산부인과학회잡지

Placental site trophoblstic tumor(PSTT) is a rare variant of trophoblastic disease. This type of trophoblastic tumor apparently exhibits different biologic behaviors as compared with choriocarcinoma. Diagnosis is made by finding a predominance of intermediate trophoblasts and absence of fetal tissue in the dilatation and currettage specimens. The intermediate trophoblastic cells produce relatively little beta-hCG and hPL and unlike other trophoblastic tumors, they are frequently resistant to chemotherapy1. But patients with metastasis frequently exhibit a progression of disease and die despite of aggressive multiagent chemotherapy. We report a case of PSTT, in which a 36-year-old woman presented with vaginal bleeding after D&CB at 14 weeks of pregnancy. Despite of blood transfusion, the patient was under shock state, and hysterectomy was done. After hysterectomy, the pathological diagnosis was PSTT confirmed by immunohistochemical study, and the result was strong positive for hPL & cytokeratin but weak positve for beta-hCG2.