Objective To evaluate different expressions of high mobility group box 1(HMGB1)and receptor for advanced glycation end products(RAGE)in placentas and their relationship with preeclampsia.Methods Fifteen early-onset pre-eclaraptic women(early-onset pre-eclampsia group),22 late-onset pre-eclamptic women(late-onset pre-eclampsia group)and 12 normotensive women(control group)in the third trimester were recruited at the Shengjing Hospital of China Medical University from March 2006 to March 2007.The localization and levels of HMGB1 and RAGE in placentas of the three groups were detected by the strept avidin biotin-peroxidose method.Results (1)Immunoreactivities to HMGB1:positive immnnostaining for HMGB1 was observed in trophoblast,macrophages,decidual cells,vascular muscle cells,endothelial cells and placental mesenchymal cells in the placentas from the pre-eclamptic women,while a low level of immunoreactivities was observed in the placentas from healthy pregnancies;the staining was observed within both the nuclei and the cytoplasm,mainly in the cytoplasm.The cytotrophoblast,especially the nuclei was extensively positive for HMGB1 in early-onset pre-eclampsia. (2)Immunoreactivities to RAGE:positive immunostaining for HMGB1 was observed in syncytiotrophoblast,macrophages and endothelial cells in the placentas from the preeclamptic women,while a low level of immunoreactivities was observed in the placentas from healthy pregnancies:the staining was in the cytoplasm and(or)cell membrane.The trophoblast was extensively positive for RAGE in early-onset pre-eclampsia.(3)Positive rate of HMGB1 expression:the expression of HMGB1 in early-onset group(73%,11/15)and late-onset group(64%,14/22)was significantly higher than that in normal group(17%,2/12;P<0.05),but no significant difference was found in early-onset group and late-onset group(P>0.05).(4)Positive rate of RAGE expression:the expression of RAGE in early-onset group(80%,12/15)and late-onset group (82%,18/22)was significantly higher than that in normal group(25%,3/12;P<0.05),but no significant difference was found in early-onset group and late-onset group(P>0.05).Conclusions The increased expression of HMGB1 and RACE in the placenta may play an important role in the pathogenesis of pre-eclampsis.The different locations may be associated with the occurrence of different onset types of pre-eclampsia.

Objective To investigate the role of hypoxia inducible factor(HIF)-prolyl hydroxylase 1 (HPHl)and factor inhibiting HIF-1(FIH-1)in placentas in the pathogenesis and development of severe pre-eclampsia.Methods RT-PCR and western blot analyses were used to detect the HPH1 and FIH-1expression levels in placentas of 34 patients with severe pre-eclampsia and 24 cases of term pregnancy (normal pregnancy group)and their correlations with symptoms were analyzed.Results (1)The HPHI mRNA and protein expression levels in placentas of severe pre-eclampsia group were 0.40±0.04 and 59.5±3.4 separately,significantly lower than those of normal pregnancy group,0.84±0.12 and 71.6±1.7(P<0.01).The FIH-1 mRNA and protein expression levels in placentas of severe pre-eclampsia group wereQ 31 ±0.05 and 45.6±2.4 separately,significantly lower than those of normal pregnancy group,0.43±0.04 and 54.9±2.1(P<0.01).(2)The mRNA and protein expression levels of HPH1 and FIH-1 in severe pre-eclampsia group were all negatively correlated with mean arterial pressure(MAP)[the Spearman correlation coefficient was-0.854(P<0.01)],urinary protein per 24 hours[the Spearman correlation coefficient was-0.936(P<0.01)1 and the occurrence of fundus oculi artery spasm[the Spearman correlation coefficient was-0.854(P<0.01)].(3)rrhe expression of HPHl mRNA in placentas of all the 58 cases WBB 0.58±0.27.higher than the expression of FIH-1 mRNA,which was 0.39±0.10.There was a positive correlation between them.The pearson correlation coefficient was 0.686(P<0.01).The expression of HPH1 protein in placentas of all the 58 cases was 64.5±6.7,higher than the expression of FIH-1,which was 49.4±5.2.There was a positive correlation between them.The Pearson correlation coefficient was 0.947(P<0.01).Conclusion The expression imbalance of HPH1 and FIH-1in palcenta may play an important role in the pathogenesis and development of severe pre-eclampsia through inhibiting HIF-1a.

Objective:To investigate the role of IL-1β in regulating human placenta-derived mesenchymal stem cells (hPMSCs) to induce the differentiation of activated peripheral blood mononuclear cells (PBMCs) into CD8 + IL-10 + T cell subset in patients with rheumatoid arthritis (RA). Methods:Serum IL-1β levels in RA patients were detected by CBA kit. hPMSCs were isolated from healthy subjects by enzyme digestion. PBMCs that were isolated from RA patients and healthy subjects by Ficoll density gradient centrifugation were activated with PHA and CD3/CD28 monoclonal antibody (McAb) and then co-cultured with hPMSCs. Flow cytometry (FCM) was used to analyze the ability of hPMSCs to induce the differentiation of activated PBMCs into CD8 + IL-10 + T cell subset with the presence of programmed death ligand 1 (PD-L1) and/or PD-L2 McAb and IL-1β. Expression of PD-L1 and PD-L2 on hPMSCs under IL-1β stimulation was analyzed by RT-PCR and FCM. Results:Serum IL-1β levels were significantly higher in RA patients than in healthy subjects. After the activation by PHA and CD3/CD28 McAb, the proportion of CD8 + IL-10 + T cells in PBMCs of RA patients was significantly lower than that of healthy subjects. Moreover, the proportion of CD8 + IL-10 + T cells significantly increased after co-cultured with hPMSCs. The ability of IL-1β-treated hPMSCs to induce activated PBMCs to differentiate into CD8 + IL-10 + T cells was significantly enhanced compared to untreated hPMSCs. IL-1β could up-regulate the expression of PD-L1 and PD-L2 on hPMSCs. Blocking the expression of PD-L1 and/or PD-L2 on hPMSCs significantly reduced the ability of hPMSCs to induce activated PBMCs to differentiate into CD8 + IL-10 + T cells. Conclusions:IL-1β enhanced the capacity of hPMSCs to induce activated PBMCs to differentiate into CD8 + IL-10 + T cells in RA patients by up-regulating the expression of PD-L1 and PD-L2.

Objective:To investigate the mechanism of IL-6 affecting the expression of CD73 on human placenta-derived mesenchymal stem cells (hPMSCs) and regulating their migration, adhesion and proliferation.Methods:Flow cytometry (FCM) and Western blot were used to analyze the effects of exogenous IL-6 or IL-6 secreted by hPMSCs on the expression of CD73 on hPMSCs. The influence of IL-6 on the phosphorylation of signal transducer and activator of transcription 3 (p-STAT3) in hPMSCs was detected by monoclonal antibody blocking test and Western blot. Real-time cellular analysis (RTCA) was used to analyze the changes in the migration, adhesion and proliferation of hPMSCs after knockdown of CD73 expression or APCP pretreatment.Results:FCM and Western blot showed that both exogenous and autocrine IL-6 from hPMSCs promoted the expression of CD73 on hPMSCs ( P<0.001, P<0.01). Moreover, CD73 expression decreased significantly with the presence of IL-6R inhibitor ( P<0.01). IL-6 could up-regulate the levels of both p-STAT3 and CD73 in hPMSCs ( P<0.05, P<0.01), while CD73 expression decreased after adding STAT3 inhibitor ( P<0.01). RTCA showed that knockdown of CD73 expression on hPMSCs significantly inhibited the adhesion and proliferation ability of hPMSCs( P<0.01, P<0.05), but promoted the migration ability of hPMSCs ( P<0.05). Similarly, inhibiting the hydrolase activity of CD73 on hPMSCs by APCP also resulted in a significant decrease in the adhesion and proliferation capacities of hPMSCs, and an increase in the migration capacity of hPMSCs ( P<0.05). Conclusions:IL-6 enhanced the expression of CD73 on hPMSCs via the JAK/STAT3 pathway, thus affecting the migration, adhesion and proliferation of hPMSCs.