Expression of glucose regulated protein 78 and the pathogenesis of gestational diabetes mellitus / 中华妇产科杂志

Objective To investigate the relationship between the expression of glucose regulated protein 78 (GRP78) in the placental trophoblast cells and the pathogenesis of gestational diabetes mellitus (GDM).Methods All the patients were recruited from Qingdao Municipal Hospital from May 2013 to May 2014.Among them, fifty women with GDM were assigned to the GDM group, and fifty healthy women were defined as the control group.All of them received cesarean section because of breech presentation, contracted pelvis, scarred uterus or on mother's demand.Real-time PCR was conducted to analyze the expression of GRP78 mRNA in the trophoblasts.Immunohistochemistry was performed to detect the localization of GRP78 protein in the placentasl trophoblast cells.Results (1) GRP78 mRNA expressed in the cytoplasm of trophoblasts of both the GDM group and the control group.The GRP78 mRNA levels in the GDM group and the control group were 15.6±0.4 and 6.0±0.7, respectively.The relative expression level of GRP78 mRNA in the GDM group was 2.6 times of that in the control group, with statistically significant difference (P＜0.01).(2) The expression of GRP78 protein was found in the cytoplasm of the trophoblasts of the GDM group.It showed in deep, light brown or yellow after staining, according to the expression degree.The expression of GRP78 protein was also found in the cytoplasm of the trophoblasts of the control group, but it mainly showed yellow color (38/50).The strong positive rate of GRP78 protein in the GDM group (96％, 48/50) was higher than that in the control group (22％, 11/50;P＜0.01).Conclusion The expression of GRP78 increased in the placental trophoblast cells of GDM patients.It might suggest that GRP78 had some effect on the pathogenesis of GDM.

Effects of transient receptor potential V6 silence on proliferation and apoptosis of trophoblasts / 中华妇产科杂志

Objective To investigate the effects of the transient receptor potential V6 (TRPV6) gene silencing on the proliferation and apoptosis of trophoblasts HTR-8/SVneo cells.Methods siRNA sequences targeting the TRPV6 gene were constructed and then transfected into HTR-8/SVneo cells mediated by liposome.The cells were divided three groups,including blank control (add the reagent of transfenction),negative control groups (transfecting nonspecific siRNA) and experimental groups (transfecting TRPV6-siRNA).Those cells in every group were collected at 24,48,72 hours after transfecting.The expression levels of TRPV6 mRNA were detected by reverse transcription (RT) PCR at different times after transfecting.The effects of siRNA on the proliferation and apoptosis of the cells were assayed by methyl thiagolyl tetragolium (MTT) and flow cytometry at different times after transfecting.Results siRNA TRPV6 transfection could inhibit the expression of TRPV6 mRNA in the HTR-8/SVneo cells.The expression was decreased with the extension of time,by 0.72 ± 0.02,0.54 ± 0.02 and 0.29 ± 0.01 after 12,48 and 72 hours of siRNA transfection as compared with the blank control and the negative control groups (P ＜0.01).The rates of proliferation inhibition were (19.29 ± 1.23) ％,(32.12 ± 1.35) ％ and (46.51 ±1.42) ％ at 24,48 and 72 hours respectively when compared with the blank control (2.12 ± 0.03)％,(2.42 ± 0.02) ％,(3.13 ± 0.04) ％ and the negative control groups (2.37 ± 0.01) ％,(2.61 ± 0.05) ％,(2.93 ± 0.03) ％ (P ＜ 0.01).The apoptosis rates of HTR-8/SVneo cells was 16.21％ at 48 hours after transfected with siRNA TRPV6,which were significantly higher than 3.27％ in the blank control and 5.34％ in the negative control groups (P ＜ 0.05).Conclusion Silenceing of TRPV6 genen could inhibit the proliferation and increase the apoptosis of extravillous trophoblas of human placenta.

Relationship of S100B protein expression and the pathogenesis of early-onset and late-onsetpreeclampsia / 中华妇产科杂志

Objective To investigate the relationship of S100B protein expression and the pathogenesis of early-onset and late-onset preeclampsia.Methods Sixty patients with preeclampsia who received caesarean section at Qingdao Municipal Hospital from October 2010 to September 2011 were enrolled in this study.Thirty cases were early-onset preeclampsia( referred as early-onset preeclampsia group,＜ 34 weeks),and the other 30 cases were late-onset preeclampsia (referred as late-onset preeclampsia group,≥34 weeks).Thirty women who received caesarean section because of pelvic structural deformities,breech presentation,macrosomia and social factors were included as the control group.The expression of S100B mRNA in the placenta was detected by reverse transcription ( RT)-PCR.The expression of S100B protein in the placenta was detected by immunohistochemistry.Results ( 1 ) S100B mRNA was expressed in the trophoblasts of preeclampsia and control groups.The expression of S100B mRNA in early-onset preeclampsia group (0.73 ±0.11 ) was significantly higher than the control group (0.58 ±0.08) and lateonset preeclampsia group (0.64 ±0.10,P ＜0.05 ).There was no significant difference between late-onset preeclampsia group and the control group ( P ＞ 0.05 ).(2) S100B protein was expressed in the plasma membrane and cytoplasm of the trophoblasts,correlated positively with the brownish yellow and brown particles inside the cells.It was expressed in all the three groups.Immunohistochemistry revealed that the expression of S100B protein in the placenta of early-onset preeclampsia group was 100％ (30/30),significantly higher than those of late-onset preeclampsia group and the control group,in which the positive rate were 70％ (21/30) and 63％ (19/30) respectively (P ＜0.05).There was no difference between late onset preeclampsia group and the control group (P ＞0.05).Conclusion Early-onset and late-onset preeclampsia may have different etiology and pathogenesis.S100B may be a factor in the pathogenesis of early-onset preeclampsia.

Expression of claudin-4 in eutopic and ectopic endometrium of women with endometriosis / 中华妇产科杂志

Objeetive To investigate the expression of claudin-4 in the eutopic and ectopic endometfium of women with endometriosis and evaluate the role of clandin-4 in the pathogenesis of endometriosis.Methods Thiny-five women with endometriosis and 35 controls were studied.Expression of elaudin-4 was investigated using immunohistochemistry,western blot and RT.PCR,respectively.Morphologic change of tight junction was also observed in different kinds of endometria.Results (1)G1andular epithelial cells of control endometrium and eutopic endometrium showed intact tight iunctions in electron micrographs,whereas the morphology of tight junctions in ovarian endometriotic tissue was disrupted and collagen bundles could be easily detected.(2)The immunohistochemical staining of claudin-4 was localized to the glandular epithelial cell membrane.Deftcient or weak staining Wag found in ovarian endometriotic tissues. In control endometrium.eutopic and ectopic endometrium of women with endometriosis,the expression of clandin-4 protein Was 89±24.84±22 and 27±14.respectively.Relative expression of claudin-4 mRNA Was 14.5±6.8,13.8±9.5 and 2.6±2.5.respectively.Expression of claudin-4 Was significantly lower in the ectopic endometriotic tissue than in the eutopic cndometrium and the control at both mRNA and protein levels(P<0.05). N0 significant difference Was foand between eutopic endometrium from women with endometriosis and control endometrium from women without endometriosis (P>0.05).Conclusion Down-regulated expression of claudin-4 might play a pathogenic role in the formation of endometriosis.

Expressions of methylthioadenosine phosphorylase gene and ornithine decarboxylase in ovarian cancer / 中国病理生理杂志

AIM:To investigate the expressions of methylthioadenosine phosphorylase(MTAP)and ornithine decarboxylase(ODC)in human ovarian cancer.METHODS:60 fresh samples of ovarian cancer were collected.The expressions of MTAP mRNA and protein were analyzed by using RT-PCR and Western blotting,respectively.ODC activity was measured by high performance liquid chromatography.RESULTS:The expression levels of MTAP mRNA and protein in ovarian cancer were lower than those of control.In 9 of the 60 samples(15%)there were absence of detectable MTAP mRNA and protein.No significant relevance was found between the expression of MTAP and clinical pathologic features.ODC activity in ovarian cancer was(3.82?1.03)U,which was higher than that of normal ovarian tissues(1.38?0.59)U.ODC activity was related with tumor grade.In MTAP-deficiency ovarian cancer tissues ODC activity was significantly increased when compared with that of MTAP-expressing ovarian cancer samples.CONCLUSION:Down-regulated MTAP expression and up-regulated ODC activity really exist in ovarian cancer.Activation of ODC resulting from MTAP deletion may be one of the pathogenetic factors of ovarian cancer.