1.Expression and significance of adiponectin in serum and placenta tissue in preeclampsia pregnancy women
Zhenyu CHEN ; Ju LI ; Jing CHEN ; Lipeng PEI
Chinese Journal of Postgraduates of Medicine 2014;37(6):1-4
Objective To analyze the expression and significance of adiponectin in serum and placenta tissue in preeclampsia pregnancy women.Methods Serum adiponectin levels were measured by Enzyme-linked immuno sorbent assay (ELISA) in 52 cases of normal pregnant women (control group),58 cases of mild preeclampsia pregnancy women (mild preeclampsia group),and 55 cases of severe preeclampsia pregnancy women (severe preeclampsia group).Insulin resistance was estimated using the homeostatic model assessment-insulin resistance index (HOMA-IR).The expression of adiponectin in placenta tissue was detected by immunohistochemical SP methods in three groups.Results Fasting glucose level in severe preeclampsia group was higher than that in control group and mild preeclampsia group [(5.56 ± 1.37) mmol/L vs.(4.55 ± 0.51),(4.68 ± 0.66) mmol/L],fasting insulin level and HOMA-IR in mild preeclampsia group and severe preeclampsia group were higher than those in control group [(14.19 ±3.42),(14.90 ±6.64) mU/L vs.(9.87 ± 1.75) mU/L,1.04 ±0.37,1.18 ±0.56 vs.0.67 ±0.21],serum adiponectin level in severe preeclampsia group was higher than that in control group and mild preeclampsia group [(15.79 ± 4.86) mg/L vs.(11.47 ±3.50),(11.92 ± 2.96) mg/L],the differences were statistical significance (P < 0.05).There was no difference of adiponectin expression levels in placenta tissue among three groups (P >0.05).There was no significant correlations between serum adiponectin level and HOMA-IR,expression levels of adiponectin in placenta tissue(P > 0.05).Conclusions Serum adiponectin levels and HOMA-IR are higher in the severe preeclampsia pregnancy women.Increased serum adiponectin levels may be an appropriate feedback regulation in preeclampsia pregnancy women.
2.Expression and purification of spindlin 1,a novel cancer related protein,and preparation of its polyclonal antibody
Lin CHEN ; Quan ZENG ; Peng ZHANG ; Jingxue WANG ; Lipeng QIN ; Yang LYU ; Xue NAN ; Wen YUE ; Xuetao PEI
Chinese Journal of Pharmacology and Toxicology 2014;(3):321-328
OBJECTIVE Toprepareapolyclonalantibodyforspindlin1protein,anovelcancer related protein,and to provide the data for a better understanding of its functions and screening tu mor. METHODS Purifiedspindlin1proteinwasinjectedintorabbitstoproducethepolyclonalantiserumafter removing glutathione S-transferase (GST)from the fusion protein spindlin 1-GST that was expressed in Escherichia coli..The antiserum was purified through the Hitrap Protein A system,and the titer of spin-dlin 1 polyclonal antibody was detected by ELISA.The specificity of the polyclonal antibody was deter-minedbyWesternblottingandimmunohistochemistry.RESULTS Thetiterofspindlin1polyclonalanti-body was 1∶2000.Western blotting detection demonstrated that the spindlin 1 polyclonal antibody recog-nized myc-spindlin 1 reco mbinant fusion protein in HeLa cells transfected with pAdeasy-myc-spindlin 1 , which also corresponded with Myc.antibody.The HeLa cells were transfected with enhanced green fluo-rescence protein (EGFP)and spindlin 1 vector(pEGFP-C3-spindlin 1 ),which was confirmed by the in-dependent GFP fluorescence assay.The results of immunohistochemistry detection with the spindlin 1 polyclonal antibody suggested that spindlin 1 was mainly expressed in the nuclei of HeLa cells.More i m-portantly,in i mmunohistoche mical assays,the spindlin 1 antibody recognized nuclear spindlin 1 expres-sioninclinicalovariancancertissues.CONCLUSION Thespecificspindlin1polyclonalantibodyispre-pared,which may be used to detect cancer-related protein spindlin 1 in HeLa cells and ovarian cancer tissues.
3.NRSF Plays a Regulatory Role in Human Insulin Gene Transcription
Qingbin LIU ; Yanhua LI ; Yinxiang YANG ; Hongfeng YUAN ; Rui ZHANG ; Lipeng QIN ; Yang L ; Sainan ZHANG ; Dijin SUN ; Lin CHEN ; Cixian BAI ; Xue NAN ; Yunfang WANG ; Xuetao PEI
Progress in Biochemistry and Biophysics 2006;0(09):-
The transcriptional repressor RE1 silencer transcription factor(NRSF/REST) is an important factor that restricts some neuronal traits in neurons.Since these traits are also present in pancreatic islet cells,NRSF-regulated genes involved in islet function are searched.A NRSE-like motif was analysed in human insulin promoter.The role of NRSE was evaluated by generating a model of insulin-secreting cells that firmly express NRSF.The presence of NRSF led to a decrease in activity of human insulin promoter by stable or transient transfection with human insulin-promoter luciferase.The predicted NRSE-like motif also confers NRSF-dependent transcriptional repression in the context of a surrogate gene promoter.Specific binding activity of NRSF/REST to the NRSE-like motif was confirmed by EMSA.Moreover,the binding activity is competed by consensus NRSE sequence.These data showed that human insulin promoter is regulated by the transcriptional repressor NRSF/REST via the NRSE-like motif.