Adipose-derived mesenchymal stem cells induced PAX8 promotes ovarian cancer cell growth by stabilizing TAZ protein.

Our previous studies have shown that the Adipose-derived mesenchymal stem cells (ADSCs) can regulate metastasis and development of ovarian cancer. However, its specific mechanism has yet to be fully revealed. In this study, an RNA-seq approach was adopted to compare the differences in mRNA levels in ovarian cancer cells being given or not given ADSCs. The mRNA level of paired box 8 (PAX8) changed significantly and was confirmed as an important factor in tumour-inducing effect of ADSCs. In comparison with the ovarian cancer cells cultured in the common growth medium, those cultured in the medium supplemented with ADSCs showed a significant increase of the PAX8 level. Moreover, the cancer cell growth could be restricted, even in the ADSC-treated group (P < .05), by inhibiting PAX8. In addition, an overexpression of PAX8 could elevate the proliferation of ovarian cancer cells. Moreover, Co-IP assays in ovarian cancer cells revealed that an interaction existed between endogenous PAX8 and TAZ. And the PAX8 levels regulated the degradation of TAZ. The bioluminescence images captured in vivo manifested that the proliferation and the PAX8 expression level in ovarian cancers increased in the ADMSC-treated group, and the effect of ADSCs in promoting tumours was weakened through inhibiting PAX8. Our findings indicate that the PAX8 expression increment could contribute a role in promoting the ADSC-induced ovarian cancer cell proliferation through TAZ stability regulation.

Increased NDRG1 expression suppresses angiogenesis via PI3K/AKT pathway in human placental cells.

OBJECTIVE: To observe whether and how N-myc downstream-regulated gene 1 (NDRG1) regulates placental angiogenesis via JEG-3 placental-derived cells. METHODS: Expression of NDRG1 in stably transfected JEG-3 cells was detected using western blot and real-time quantitative polymerase chain reaction. Angiogenesis was examined by tube formation assay. The levels of placental growth factor (PLGF) and soluble fms-like tyrosine kinase-1 (sFlt-1) were examined using enzyme-linked immunosorbent assay. The expression of vascular endothelial growth factor (VEGF), PI3K, and AKT was examined by western blot. The relationship between PI3K and NDRG1 was detected by co-immunoprecipitation. RESULTS: NDRG1 was significantly down-regulated at both the mRNA and protein level by lentivirus (Lv)-NDRG1-shRNA (P < 0.001), whereas it was significantly up-regulated by Lv-NDRG1 (P < 0.001). NDRG1 knockdown significantly increase the expression of PLGF and VEGF in JEG-3 cells (P < 0.001), while NDRG1 knockdown significantly reduced the secretion of sFlt-1 (P < 0.001). NDRG1 was specific bound to PI3K, and NDRG1 knockdown significantly up-regulated the expressions of PI3K and AKT in JEG-3 cells (P < 0.001). CONCLUSION: NDRG1 suppresses angiogenesis in preeclampsia, and the PI3K/AKT signaling pathway may be involved in the regulation of angiogenesis by NDRG1.

Genetic susceptibility analysis of GCLC rs17883901 polymorphism to preeclampsia in Chinese Han women.

Preeclampsia (PE) is a specific obstetric disorder that may result in maternal and neonatal morbidity and mortality. Increasing evidence has been indicated that some candidate genes related to oxidative stress, such as glutamate-cysteine ligase, catalytic subunit (GCLC), glutamate-cysteine ligase, modifier subunit (GCLM), involve in the pathogenesis of PE. After the genetic contribution of GCLC rs17883901 polymorphism was analyzed by TaqMan allelic discrimination real-time PCR in 1001 PE patients and 1182 normal pregnant women, a case-control association analysis was performed. Although no statistical difference was found in genetic distribution of rs17883901 in GCLC between PE and control group (χ2 = 2.201, p = .333 by genotypic, χ2 = 0.524, p = .469, OR = 0.932, 95%CI = 0.771-1.128 by allelic), significant differences in the genotypic frequencies were investigated between mild PE group (χ2 = 6.999, p = .030) or late-onset PE group (χ2 = 6.197, p = .045) and control group. Furthermore, when dividing the mild PE patients, the late-onset PE patients and the controls into TT/CT + CC, TT + CT/CC, and TT/CC subgroups, we found statistical differences between mild PE and controls (TT/CT + CC:χ2 = 5.132, p = .023, OR = 2.948, 95%CI = 1.107-7.854; TT/CC:χ2 = 4.564, p = .033, OR = 2.793, 95%CI = 1.046-7.460) as well as late-onset PE and controls (TT/CT + CC:χ2 = 4.043, p = .044, OR = 2.248, 95%CI = 1.000-5.055). This is the first study to indicate GCLC rs17883901 polymorphism may be associated with a risk of mild PE and late-onset PE in Chinese Han women. However, additional well-designed studies with multi-ethnic and large-scale samples should be performed to validate our results.

Reduced ELABELA expression attenuates trophoblast invasion through the PI3K/AKT/mTOR pathway in early onset preeclampsia.

INTRODUCTION: Early onset preeclampsia is linked to abnormal trophoblast invasion, leading to insufficient recasting of uterine spiral arteries and shallow placental implantation. This study investigated ELABELA (ELA) expression and its involvement in the pathogenesis of early onset preeclampsia. METHODS: We used immunohistochemistry, quantitative PCR and Western blot to calculate ELA levels in the placentas. Transwell assays were utilize to assess the invasion and migration of trophoblastic Cells. Western blot was used to identify the concentrations of vital kinases in PI3K/AKT/mTOR pathways and invasion-related proteins in trophoblast cells. RESULTS: ELA was expressed in villous cytotrophoblasts and syncytiotrophoblasts in placental tissue. Compared with the normal pregnancies, ELA mRNA and protein expression was significantly reduced in early onset preeclampsia placentas. In the HTR-8/SVneo cells, when ELA was knocked down, the invasion and migration capability of cells decreased significantly, with MMP2 and MMP9 expression downregulated and the expression of important kinases in the PI3K/AKT/mTOR pathways being significantly decreased compared to the control group. Overexpression of ELA was on the contrary. Besides, while PI3K was blocked, the invasion and migration capability of HTR-8/SVneo cells and the expression of key kinases in PI3K/AKT/mTOR pathways were decreased significantly. DISCUSSION: ELA stimulates the invasion and migration of trophoblastic cells through activation of downstream PI3K/AKT/mTOR pathway and is complicit in early onset preeclampsia pathogenesis. Our research offers a potential novel treatment for PE.

Downregulation of receptor tyrosine kinase-like orphan receptor 1 in preeclampsia placenta inhibits human trophoblast cell proliferation, migration, and invasion by PI3K/AKT/mTOR pathway accommodation.

INTRODUCTION: Invasive deficiency of the trophoblast and poor remodeling of the uterine spiral arteries were probably the primary pathogenesis causes of preeclampsia (PE). The expression of receptor tyrosine kinase-like orphan receptor 1 (ROR1) during embryogenesis had been previously confirmed and was closely related to the function of tumor cells, which was similar to the characteristics of trophoblasts. In this work, we investigated the expression profile of ROR1 in preeclampsia placentas and the functional role of ROR1 in trophoblast cells, as well as the associated molecular mechanisms. METHODS: The localization expression of ROR1 in the placenta was detected by immunohistochemistry in 20 cases of normal term pregnancy, preterm delivery, late-onset severe PE, and early-onset severe PE, respectively. The expression levels were determined by fluorescence quantitative PCR and Western blot. The influence of ROR1 on trophoblast proliferation, migration, invasion, and potential regulatory pathways was evaluated in HTR-8/SVneo cell lines by transient transfection methods. RESULTS: The levels of ROR1 in the placental tissues in PE were significantly lower than those in normal term pregnancy and preterm delivery. Moreover, the expression levels of ROR1 in early-onset severe PE were significantly lower than those in its late counterparts. ROR1 overexpression increased cell proliferation, migration, and invasion of HTR-8/SVneo cells, whereas its silencing had the opposite effect. Meanwhile, the phosphorylation levels of critical kinases in the PI3K/AKT/mTOR pathways were increased by ROR1 overexpression, whereas they were decreased by the silencing of ROR1. CONCLUSION: ROR1 might be involved in the development of PE through regulating trophoblast viability, migration, and invasion by PI3K/AKT/mTOR signaling pathway.

Decreased Filamin b expression regulates trophoblastic cells invasion through ERK/MMP-9 pathway in pre-eclampsia.

OBJECTIVES: The purpose of this study was to investigate the expression of Filamin b in the placental placenta of patients with early or late onset pre-eclampsia (PE) and its potential effects on the pathophysiology of the disease. METHODS AND METHODS: Immunohistochemistry staining, western blot assays and real time PCR were used to detect the expression level of FLN-b. The expression levels of MMP-2, MMP-9 and ERK1/2 proteins from control and FLN-b-silenced JEG-3 cells were also detected by western blot and JEG-3 cell invasion. RESULTS: Compared with normal term pregnancies placentas, the FLN-b expression was significantly lower than that of women with PE, its level in late-onset PE is lower than in early-onset PE. In FLN-b-silenced JEG-3 cells, the protein levels of MMP-2, MMP-9 and phosphorylated ERK1/2 decreased markedly and the number of cells penetrating through the transwell chamber membrane is also greatly reduced. CONCLUSIONS: Down-regulation of FLN-b inhibits the ERK/MMP-2 and MMP-9 pathways, leading to trophoblastic invasion disorders in the PE placenta.

STAT3 activation by IL-6 from adipose-derived stem cells promotes endometrial carcinoma proliferation and metastasis.

Endometrial cancer is the most common gynaecological cancer, and its incidence is increasing. Obesity is a well-recognized risk factor for endometrial cancer, and the mechanisms by which adipose tissue influences tumour development remain controversial. In this study, we examined the high IL-6 level in the ADSCs supernatant following treatment of endometrial cancer cell CM. Then, the activation of STAT3, a major tumourigenic IL-6 effector, was examined in ADSCs CM treated endometrial cancer cells. Conditioned ADSC medium was used to stimulate endometrial cancer cell growth in vitro. Similar to IL-6, ADSC-conditioned medium significantly promoted endometrial cancer growth and invasion. Furthermore, siRNA-mediated STAT3 inhibition in endometrial cancer cells decreased the ADSC-mediated promotion of cell proliferation and invasion. In addition, a subcutaneous nude mouse model of endometrial cancer was established to monitor the tumour-promoting effect of ADSCs. ADSC-conditioned medium promoted tumour growth, and STAT3 inhibition attenuated this effect. Based on these data, ADSCs promote endometrial cancer progression by the STAT3 signalling pathway.

Progesterone inhibited endoplasmic reticulum stress associated apoptosis induced by interleukin-1ß via the GRP78/PERK/CHOP pathway in BeWo cells.

AIM: Pre-eclampsia (PE) is a pregnancy complication characterized by new onset maternal hypertension and proteinuria. Its underlying mechanisms are unclear. This study investigated the relationship between progesterone and endoplasmic reticulum stress (ERS) associated apoptosis induced by interleukin (IL)-1ß via the glucose regulated protein 78 (GRP78)/protein kinase RNA-like endoplasmic reticulum kinase (PERK)/C/EBP-homologous protein (CHOP) pathway in BeWo cells. METHODS: Venous blood and placental tissues were collected from PE patients, normal pregnancy and preterm delivery cases, respectively. Progesterone serum levels were detected by enzyme-linked immunosorbent assay and ERS-related protein expression in placentas was examined by immunohistochemistry, reverse transcriptase-polymerase chain reaction and Western blot. BeWo cells were stimulated by IL-1ß to induce ERS associated apoptosis in vitro. The apoptotic rate was measured by flow cytometry. The mechanism of progesterone acting on IL-1ß induced ERS associated apoptosis was investigated by reverse transcriptase-polymerase chain reaction, Western blot and PERK small interfering RNA, with RU486 used as a receptor inhibitor. RESULTS: PE patients exhibited decreased serum levels of progesterone and activated ERS and increased ERS-related protein expression. IL-1ß could induce ERS and associated cell apoptosis by activating the GRP78/PERK/CHOP signal pathway, which could be inhibited by progesterone. PERK could be upregulated and phosphorylation activated in ERS. The protective effects of progesterone could be attenuated by RU486. CONCLUSION: IL-1ß could induce ERS associated cell apoptosis by activating the GRP78/PERK/CHOP signal pathway in BeWo cells and may play an important role in PE occurrence. Progesterone levels were decreased in patients with PE and seemed to have a protective effect by inhibiting ERS associated cell apoptosis.

Accumulation of Advanced Glycation End Products Involved in Inflammation and Contributing to Severe Preeclampsia, in Maternal Blood, Umbilical Blood and Placental Tissues.

OBJECTIVE: To investigate the expression of advanced glycation end products (AGEs) and the receptor for AGE (RAGEs) in maternal blood, umbilical blood and placental tissues in women with severe preeclampsia (sPE) as well as any association with inflammatory processes. METHODS: The expressions of AGEs, RAGE, tumor necrosis factor-alpha (TNF)-α and vascular cell adhesion molecule-1 (VCAM)-1 in placental tissues were measured using immunohistochemistry. The levels of AGEs, RAGE, TNF-α and VCAM-1 in maternal blood, umbilical blood and placental extracts were assessed using enzyme-linked immunosorbent assays. Placental RAGE, TNF-α and VCAM-1 mRNA expression levels were determined by PCR. Placental AGEs, RAGE, TNF-α and VCAM-1 protein levels were determined by western blotting. RESULTS: The levels of AGEs, TNF-α and VCAM-1 in the maternal tissues and umbilical blood were significantly higher in the sPE group than in the normal pregnancy (NP) controls (p < 0.05). The serum level of sRAGE in the umbilical blood was lower in the sPE group than in the NP controls (p < 0.05), while sRAGE was higher in the maternal blood of sPE than in the NP (p < 0.05). The maternal serum levels of AGEs were positively correlated with that of TNF-α and VCAM-1 in the maternal blood. There were no correlations between the levels of RAGE, TNF-α or VCAM-1 in maternal blood or umbilical serum. There were no correlations between the levels of sRAGE and TNF-α or VCAM-1 in maternal blood or umbilical serum. The levels of AGEs were positively correlated with those of TNF-α and VCAM-1 in placental lysates. CONCLUSION: AGEs and RAGE appear to act as important mediators in regulating the inflammatory pathways of preeclampsia.

Autophagy protects against oxidized low density lipoprotein-mediated inflammation associated with preeclampsia.

INTRODUCTION: Inflammatory responses play an important role in the pathogenesis of preeclampsia. Recently, the anti-inflammatory role played by autophagy has drawn increasing attention. Our aim was to investigate variations in autophagy in preeclampsia and protection against oxidized low-density lipoprotein (oxLDL)-mediated inflammation by autophagy. METHODS: We used immunohistochemistry, immunofluorescence, quantitative real-time PCR, and western blotting to analyze the expression of autophagy proteins (beclin-1 and LC3II/LC3I) in preeclampsia placentas and in JEG-3 cells treated with oxLDL and rapamycin. RESULTS: We found a decreased level of autophagy proteins in preeclampsia placentas, and oxLDL did not induce autophagy in JEG-3 cells. Furthermore, when cells were pretreated with rapamycin, autophagy was activated and expression of inflammatory factors (tumor necrosis factor-α and interleukin-6) induced by oxLDL was downregulated. CONCLUSION: We conclude that impaired autophagy in preeclampsia has potential to decrease trophoblast protection from oxidative and inflammatory stress, thereby contributing to the pathogenesis of preeclampsia.

[Correlation of the expressions of advanced glycation end products and its receptor in serum and placenta with the pathogenesis of preeclampsia].

OBJECTIVE: To investigate the correlation of the expressions of advanced glycation end products (AGE) and the receptor for advanced glycation end products (RAGE) in serum and placenta with the pathogenesis of preeclampsia. METHODS: From December 2013 to June 2014, 32 women with severe preeclampsia who received cesarean section in the Affiliated Hospital of Qingdao University were recruited in the study, defined as the severe preeclampsia group. 30 healthy pregnant women who received cesarean section in the same hospital were recruited as the control group. ELISA was used to measure the maternal serum AGE, soluble receptor for advanced glycation end products (sRAGE) and tumor necrosis factor-α (TNF-α) in these women. Furthermore, ELISA was also used to measure AGE and TNF-α in the placenta. The localizations of AGE and RAGE protein in placentas were detected by immunohistochemical SP method. RAGE and TNF-α mRNA expression in placentas were measured by real-time quantitative PCR. AGE, RAGE and TNF-α protein expression in placentas were measured by western blot, respectively. RESULTS: (1) The serum levels of AGE, sRAGE and TNF-α in the severe preeclampsia group were (538 ± 75), (367 ± 86) and (322 ± 40) ng/L, respectively. They were significantly higher than those in the control group [(454 ± 50), (286 ± 35) and (270 ± 35) ng/L, respectively] (P < 0.05). The levels of AGE showed positive correlation with the levels of TNF-α (r = 0.588, P < 0.05), while the levels of sRAGE showed no correlation with TNF-α (r = -0.041, P > 0.05). (2) In the severe preeclampsia group, the levels of AGE and TNF-α in placentas were (500 ± 82) and (334 ± 57) ng/L, which were higher than those in the control group [(431 ± 74) and (263 ± 46) ng/L, respectively] (P < 0.05). The levels of AGE showed positive correlation with the levels of TNF-ɑ (r = 0.406, P < 0.05). (3) AGE and RAGE protein mainly located in the syncytiotrophoblasts, macrophages and vascular endothelial cells in the placentas of the two groups. AGE expressed mainly in the cytoplasm, and RAGE expressed in the cytoplasm and cell membranes. (4) RAGE and TNF-α mRNA expression in the placentas of the severe preeclampsia group were 12.6 ± 4.6 and 10.4 ± 2.4, which were significantly higher than those in the control group (0.9 ± 0.4 and 3.5 ± 0.9, P < 0.01). (5) The expressions of AGE, RAGE and TNF-α protein in placentas of the severe preeclampsia group were 0.68 ± 0.06, 0.82 ± 0.08 and 0.76 ± 0.08. All were significantly higher than those of the control group (0.46 ± 0.05, 0.42 ± 0.09 and 0.52 ± 0.07; P < 0.01). CONCLUSIONS: The levels of AGE and RAGE in serum and placentas elevated in the severe preeclampsia group, and the expression of TNF-α also elevated. These indicated that AGE and RAGE might be involved in the systemic inflammatory response and local inflammatory response in placentas, and then caused the preeclampsia.

Inhibition of lectin-like oxidized low-density lipoprotein receptor 1 protects against plasma/hypoxia-mediated trophoblast dysfunction associated with preeclampsia.

BACKGROUND: Preeclampsia (PE) is associated with oxidative stress in the maternal circulation and placenta. This study aimed to determine if inhibition of lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1) gives protection against oxidative stress-mediated trophoblast dysfunction. METHODS: Plasma and placenta samples were obtained from 106 women with PE and 106 women with normal pregnancy (NP). Oxidized low-density lipoprotein (oxLDL) and soluble LOX-1 levels were determined by enzyme-linked immunoassay. Placental LOX-1 expression was determined by western blotting. Trophoblasts were subjected to hypoxia and treated with pooled plasma from patients with PE. Expression levels of placenta growth factor (PIGF) and the soluble form of the PIGF receptor (sFlt-1) in trophoblasts were determined. RESULTS: Plasma concentrations of oxLDL and sLOX-1 were significantly over-expressed and LOX-1 protein expression in the placenta was significantly increased in PE patients compared with matched NP controls (both p < 0.05). Exposure of trophoblasts to hypoxia and pooled PE plasma induced overexpression of sFlt-1 and downregulation of PIGF. These effects were inhibited by the LOX-1 inhibitor TS20. CONCLUSION: LOX-1 accumulation may contribute to the pathogenesis of PE by promoting sFlt-1 production in trophoblasts, suggesting that oxidative stress may be an important mediator regulating angiogenic pathways in trophoblasts.

[Correlation of lipocalin-2 and its receptor expressions with preeclampsia].

OBJECTIVE: To research the correlation of the expressions of lipocalin-2 (LCN-2) and its receptor (NGALR) in serum and placenta with preeclampsia. METHODS: From Dec.2010 to Apr.2011, 64 women with preeclampsia who delivered in Affiliated Hospital of Qingdao University Medical College were recruited in the study, including 26 women with moderate preeclampsia (MPE group) and 38 women with severe preeclampsia (SPE group). Twenty-five healthy pregnant women were taken as control group. LCN-2 and NGALR mRNA and protein expression in placenta were measured by reverse transcription-PCR (RT-PCR) and western blot, respectively. RESULTS: (1) The serum levels of LCN-2 in MPE group and SPE group [(58 ± 20), (90 ± 18) µg/L] were significantly higher than that in control group [(19 ± 6) µg/L, P < 0.01]; the serum LCN-2 level in SPE group was significantly higher than that in MPE group (P < 0.01). (2) LCN-2 mRNA expression in placenta in MPE group and SPE group (0.55 ± 0.14, 0.61 ± 0.14) were both significantly higher than that in control group (0.28 ± 0.16, P < 0.01); LCN-2 protein expression in placenta of MPE group and SPE group (2.2 ± 0.4, 2.4 ± 0.5) were also significantly higher than that in control group (1.4 ± 0.4, P < 0.01), no significant difference was found between MPE group and SPE group (P > 0.05). (3) No significant difference was found in the expressions of NGALR mRNA in placenta among MPE group, SPE group and control group (0.46 ± 0.11, 0.46 ± 0.14, 0.45 ± 0.15, P > 0.05). (4) NGALR protein expressions in MPE group, SPE group and control group were 2.7 ± 0.8, 3.0 ± 0.9, and 2.7 ± 0.9, and there were no significant difference among these three groups (P > 0.05). (5) In preeclampsia, serum LCN-2 level significant associated with 24 hours total urinary protein and uric acid (r = 0.565, 0.476, P < 0.01). LCN-2 serum level were not associated with systolic pressure and diastolic pressure (P > 0.05); there were no association with the expressions LCN-2 mRNA and protein in placenta (P > 0.05). CONCLUSIONS: Serum LCN-2 level is closely related to the progress of preeclampsia. Increasing expression of LCN-2 in placenta may be a compensatory response to preeclampsia.

[Association between plasma levels of soluble leukocyte differentiation antigens CD40/CD40 ligand and kidney damage in preeclamptic patients].

OBJECTIVE: To investigate the variance levels of plasma soluble leukocyte differentiation antigens CD40 (sCD40) and soluble CD40 ligand (sCD40 L) in preeclamptic patients with renal damage and its relationship. METHODS: A total of 63 pregnant women attended the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College between August 2008 and June 2010. In the present study included 28 pregnant women with mild preeclampsia and 35 patients with severe preeclampsia. Thirty matched normotensive pregnant women were enrolled in the study as the control group. Expression of sCD40 and sCD40 L were determined by ELISA. At the same time, the blood routine, C reaction protein (CRP), urine routine, 24 hours urine protein excretion, and serum uric acid (UA), creatinine (Cr), blood urea nitrogen (BUN) were measured. The correlation analysis was performed between the sCD40/sCD40 L and the blood biochemical indexes in 3 groups. RESULTS: (1) The median levels of CRP in severe preeclampsia (10.8 mg/L) and mild preeclampsia group (7.1 mg/L) are significantly higher than that of control group (3.3 mg/L, P < 0.05); The level of CRP in severe preeclampsia group was also higher than that of mild preeclampsia group (P < 0.05). The median gestational age at delivery in severe preeclampsia (32.5 weeks) was significantly less than that of mild preeclampsia group (37.2 weeks) and normal group (38.6 weeks, P < 0.05). However no significant differences were observed between mild preeclampsia group and normal group (P > 0.05). The platelet count in severe preeclampsia (132 × 109/L) was significantly less than those of mild preeclampsia group (212 × 109/L) and normal group (216 × 109L, P < 0.01), but no significant differences were observed in blood platelet amount between mild preeclampsia group and normal group (P > 0.05). There was no significant difference in hemoglobin level and white blood cell in three groups (P > 0.05). (2) The sCD40 plasma concentration in severe, mild preeclampsia and normal group was 133.6, 126.5 and 90.7 ng/L, respectively. The sCD40 L plasma concentrations were 12.5, 10.4 and 4.4 ng/L respectively in the 3 groups. 24 hours urinary protein quantitative was 4.5 g/d, 0.8 g/d and 0 in the 3 groups respectively. And the UA level was 486 µmol/L, 289 µmol/L and 162 µmol/L. In the above three groups, the monitoring indicators were significantly higher in women with severe preeclampsia group compared with mild preeclampsia and control groups (P < 0.01), and there were also higher in mild preeclampsia group than that in control groups (P < 0.01). The level of plasma Cr (89 µmol/L) and BUN (5.32 mmol/L) in severe preeclampsia group were higher than those of mild preeclampsia group (66 µmol/L and 4.49 mmol/L) and control group (57 µmol/L and 3.32 mmol/L, P < 0.05). There was no significant difference between mild preeclampsia group and normal group (P > 0.05). (3) The correlation analysis indicated that the level of sCD40 has a positive correlation with 24 hours urinary protein quantitative (r = 0.434, P < 0.05), also significant positive correlation (r = 0.536, 0.528, P < 0.01) between the level of sCD40 and UA or CRP in women with preeclampsia. There was no significant correlation between the level of sCD40 and systolic blood pressure, diastolic blood pressure, delivery gestational age, Cr, BUN, and platelet count (r = 0.135, 0.183, -0.133, 0.190, 0.167, -0.221, all P > 0.05). There were positive correlation between the level of sCD40 L and 24 hours urine protein excretion, either UA or CRP (r = 0.591, 0.445, 0.539, all P < 0.01). No significant correlation was found between sCD40 L and systolic blood pressure, diastolic blood pressure, delivery gestational age, Cr, BUN, and platelet count (r = 0.178, 0.212, -0.292, 0.144, 0.135, -0.273, all P > 0.05). There was significant positive correlation between plasma sCD40 and sCD40 L (r = 0.707, P < 0.01). There was no relationship between the level of sCD40, sCD40 L and the blood biochemical indexes in normotensive pregnant women (P > 0.05). CONCLUSIONS: The plasma concentrations of sCD40 and sCD40 L are significantly higher in pregnant women with preeclampsia compared with the control, which may be involved in the development of preeclampsia and contribute to the kidney damage. The variance levels of sCD40 and sCD40 L may be also related to the severity of preeclampsia.

[Predictive value of serum soluble fms-like tyrosine kinase 1 concentration in preeclampsia at second trimester].

OBJECTIVE: To explore the predictive value of serum soluble fms-like tyrosine kinase-1 (sFlt-1) and vascular endothelia growth factor (VEGF) levels in preeclampsia at second trimester. METHODS: Serum sFlt-1, VEGF concentrations were determined in 172 initial normal pregnant women at 26 - 28 gestation week. The outcomes of pregnancies were followed. In a cohort of 172 pregnant women, 16 cases of preeclampsia were developed (preeclampsia group), and 156 cases were with no complication (control group). RESULTS: The serum levels of sFlt-1 in preeclampsia group (11.4 +/- 6.2) microg/L were significantly higher than that in control group (4.5 +/- 2.1) microg/L (P < 0.01). The serum levels of sFlt-1 in precelampsia women with the onset before 32 gestation week and fetal growth retardation, (14.0 +/- 6.8) microg/L, (14.4 +/- 6.7) microg/L were significantly higher than that in women with the onset after 32 gestation week and with no fetal growth retardation (9.0 +/- 4.1) microg/L, (8.9 +/- 4.0) microg/L, respectively (P < 0.05). There was no significant difference in the serum levels of VEGF between preeclampsia group and control group. A sFlt-1 cutoff value of 8.75 microg/L at 26 - 28 gestation week yielded a sensitivity of 87.5%, specificity of 97.4%, positive predictive value of 80.0%, negative predictive value of 88.5%, respectively, for subsequent onset of preeclampsia. CONCLUSION: Maternal serum sFlt-1 concentration at second trimester can be used as an early predictive marker of preeclampsia.

[Expression and significance of soluble fms-like tyrosine kinase 1 in preeclampsia placenta].

OBJECTIVE: To investigate the expression and significance of serum soluble fms-like tyrosine kinase 1 (sFlt-1) in preeclampsia placenta. METHODS: Immunohistochemistry and semi-quantitative RT-PCR were used to investigate the expression of sFlt-1 and sFlt-1 mRNA in placenta of 30 women with preeclampsia (including mild and moderate preeclampsia 11 cases, severe preeclampsia 19 cases) and 45 normal pregnant women (including first trimester pregnancy 18 cases, 2nd trimester pregnancy 12 cases, and 3rd trimester pregnancy 15 cases). The levels of vascular endothelial growth factor (VEGF) and sFlt-1 in serum were detected by enzyme linked immunosorbent assay. RESULTS: (1) The expression of sFlt-1 mRNA was significantly higher in preeclampsia group (0.90 +/- 0.11) compared with the third trimester group (0.80 +/- 0.06; P < 0.01), and was higher in the severe preeclampsia group (0.93 +/- 0.12) than that in the mild group (0.85 +/- 0.05; P < 0.05). (2) The mean density of sFlt-1 in preeclampsia placenta (0.156 +/- 0.008) was significantly higher than that in the third trimester group (0.143 +/- 0.009, P < 0.01), and was higher in the severe group (0.159 +/- 0.008) than in the mild group (0.151 +/- 0.005; P < 0.05). (3) The level of VEGF in serum of the preeclampsia group (19.3 +/- 2.9) ng/L was significantly lower than that of the third trimester group [(30.2 +/- 3.1) ng/L, P < 0.01]. The level of sFlt-1 in serum of the preeclampsia group (30.2 +/- 13.7) microg/L was significantly higher than that of the third trimester group [(7.4 +/- 3.1) microg/L, P < 0.01]. (4) There was significant correlation between the serum level of sFlt-1 and the expression of sFlt-1 mRNA or sFlt-1 in placenta of normal pregnancy group (r = 0.314, P < 0.05; r = 0.439, P < 0.01) and preeclampsia groups (r = 0.372, r = 0.383; P < 0.05). CONCLUSION: Upregulation of sFlt-1mRNA and excess expression of sFlt-1 in placenta may induce higher level of sFlt-1 in serum, which may be involved in the pathophysiological processes of preeclampsia.

[Expression of matrix metalloproteinase-9 and tumor necrosis factor-alpha in the placenta of patients with pregnancy-induced hypertension].

OBJECTIVE: To investigate the role of matrix metalloproteinase-9 (MMP-9) and tumor necrosis factor-alpha (TNF-alpha) in the pathogenesis of pregnancy-induced hypertension (PIH). METHODS: The placenta samples were collected from 57 patients with PIH and 24 normal pregnant women. Immunohistochemical staining was used to examine positive expression of MMP-9 and TNF-alpha in these samples. RESULTS: The positive expression of MMP-9 in the normal group was significantly higher than that in patients with moderate or severe PIH (P<0.01), while no significant difference was found between normal pregnant women and patients with mild PIH (P>0.05), and expression decreased drastically with the progression of PIH (P<0.05). The expression of TNF-alpha in the patients with mild, moderate and severe PIH groups were significantly higher than that in the normal group (P<0.05, P<0.01 and P<0.01, respectively), and the differences were also significant between the PIH groups of different severities (P<0.01). No correlation was found between MMP-9 and TNF-alpha expressions in the normal and mild PIH groups (r=0.287, P>0.05; r=0.382, P>0.05), in the patients with moderate and severe PIH, MMP-9 expression showed inverse correlation with TNF-alpha expression (r=-0.563, P<0.05; r=-0.681, P<0.01). CONCLUSION: Lowerd MMP-9 expression in syncytiotrophoblast might result in defective nidation in the placenta where local ischemia and hypoxia cause abnormal TNF-alpha elevation, which might be one of the important factors inducing PIH.