MicroRNA-30a-3p is overexpressed in the placentas of patients with preeclampsia and affects trophoblast invasion and apoptosis by its effects on IGF-1.

OBJECTIVE: Preeclampsia (PE) affects many women globally and remains a primary cause of neonatal and maternal morbidity and mortality. Aberrant placental microRNA (miRNA) expression might be associated with PE. Previously, 33 PE-related miRNAs, 11 up-regulated and 23 down-regulated, were detected in placentas of women with severe PE when compared with those of normal patients. One of the most up-regulated miRNAs in PE is miR-30a-3p. The predicted target of it is insulin-like growth factor 1 (IGF-1), which has been reported to have a relatively low expression level in PE patients. This study was conducted to determine the aberrant increased of miR-30a-3p in the placentas of women with preeclampsia and to elucidate the target and function of it in trophoblast cells. STUDY DESIGN: miR-30a-3p expression in placenta tissues was compared between women with preeclampsia (n = 25) and normal pregnant women (n = 20). The miRNA target was studied by in silico and functional assay. The effects of the miRNA were verified by apoptosis assay and invasion assay in the trophoblast cell line. RESULTS: miR-30a-3p was increased significantly in the placenta of women with preeclampsia when compared to those with normal pregnancies. Luciferase assay confirmed direct regulation of miR-30a-3p on the expression of IGF-1. Forced expression of miR-30a-3p suppressed IGF-1 protein expression in the HTR-8/SVneo cells. The functional assay suggests that the over-expression of miR-30a-3p alter the invasive capacity of JEG-3 cells and induce the apoptosis of HTR-8/SVneo cells (Figure). CONCLUSION: Expression of miR-30a-3p was significantly increased in the placentas of patients with preeclampsia. miR-30a-3p might be involved in the pathogenesis of preeclampsia by targeting IGF-1 and regulating the invasion and apoptosis of trophoblast cells.

FAM3A Protects Against Glutamate-Induced Toxicity by Preserving Calcium Homeostasis in Differentiated PC12 Cells.

BACKGROUND/AIMS: Stroke is the leading cause of adult disability, and glutamate-induced dysregulation of intracellular Ca2+ homeostasis is a key mechanism. FAM3A is the first member of the family with sequence similarity 3 (FAM3) gene family, and its biological function remains largely unknown. We have recently reported that FAM3A exerts protective effects against oxidative stress and mitochondrial dysfunction in HT22 cells. METHODS: Here, we investigated the protective effects of FAM3A using a glutamate-induced neuronal injury model in nerve growth factor (NGF)-differentiated PC12 cells. The protective effects were determined by measuring lactate dehydrogenase (LDH) release, apoptosis and mitochondrial oxidative stress. Ca2+ imaging was performed to detect changes in intracellular Ca2+ concentration in PC12 cells. The related molecular mechanisms were investigated by fluorescence staining, coimmunoprecipitation (Co-IP) and western blotting. RESULTS: Upregulation of FAM3A by lentivirus transfection markedly decreased LDH release, inhibited apoptosis and reduced mitochondrial oxidative stress, which were accompanied by alleviated intracellular Ca2+ levels as measured by calcium imaging. The results of western blotting showed that FAM3A significantly decreased the surface expression of metabotropic glutamate receptor 1/5 (mGluR1/5), with no effect on the expression of N-methyl-d-aspartic acid receptor (NMDAR) or α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor (AMPAR) subunits. FAM3A overexpression also inhibited the intracellular Ca2+ release mediated by mGluR1/5 and inositol 1,4,5-trisphosphate receptor (IP3R), but not the ryanodine receptor (RyR). In addition, FAM3A significantly attenuated the store-operated calcium entry (SOCE) induced by thapsigargin (Tg), but the expression of SOCE-related proteins was not altered. The results of coimmunoprecipitation (Co-IP) showed that FAM3A disrupted the interaction of stromal interaction molecule 1 (STIM1) with Orai1 triggered by glutamate. CONCLUSION: These results suggest that the upregulation of FAM3A protects against glutamate-induced dysfunction of Ca2+ homeostasis not only by inhibiting mGluR1/5-dependent endoplasmic reticulum (ER) Ca2+ release, but also by attenuating SOCE mediated by the STIM1-Orai1 interaction.

FAM3A Protects HT22 Cells Against Hydrogen Peroxide-Induced Oxidative Stress Through Activation of PI3K/Akt but not MEK/ERK Pathway.

BACKGROUND/AIMS: Oxidative stress-induced cell damage is involved in many neurological diseases. FAM3A is the first member of family with sequence similarity 3 (FAM3) gene family and its biological function remains largely unknown. METHODS: This study aimed to determine its role in hydrogen peroxide (H2O2) induced injury in neuronal HT22 cells. The protective effects were measured by cell viability, lactate dehydrogenase (LDH) release and apoptosis, and oxidative stress was assayed by reactive oxygen species (ROS) generation, ATP synthesis and lipid peroxidation. By using selective inhibitors, the involvement of PI3K/Akt and MEK/ERK pathways were also investigated. RESULTS: The results of fluorescence staining revealed that H2O2 significantly decreased the expression of FAM3A protein, which was shown to be subcellularly located in mitochondria. Up-regulation of FAM3A by lentivirus transfection markedly increased cell viability and decreased LDH release after H2O2 treatment. The anti-apoptotic activity of FAM3A was demonstrated by the reduced mitochondrial cytochrome c release, decreased activation of caspase-3 and the results of flow cytometry. Overexpression of FAM3A attenuated intracellular ROS generation and loss of ATP production induced by H2O2, and subsequently inhibited lipid peroxidation. In addition, overexpression of FAM3A significantly increased the activation of Akt and ERK in H2O2 injured HT22 cells. By using Akt and ERK specific inhibitors, we found that inhibition of PI3K/Akt, but not MEK/ERK pathway, partially prevented FAM3A-induced protection against H2O2. CONCLUSION: These results suggest that FAM3A has protective effects against H2O2-induced oxidative stress by reducing ROS accumulation and apoptosis, and these protective effects are dependent on the activation of PI3K/Akt pathway.

[Early glucose challenge test in pregnant women with risk factors of GDM in China].

OBJECTIVE: To understand the current status and clinical relevance of early (<24 weeks) glucose challenge test (GCT) in pregnant women with risk factors of gestational diabetes mellitus (GDM) in China. METHODS: Data from the survey of incidence of GDM in China were re-analyzed. The incidence of abnormal glucose metabolisms and the rate of early GCT in all women were calculated according to different numbers of risk factors. Sixteen risk factors were included in the survey. However, 4 independent risk factors were considered separately in this re-analysis. The ADA criteria for GDM diagnosis were applied. RESULTS: A total of 16 286 pregnant women were included in this analysis and 64.3% (10 468) presented with at least one risk factor. The incidence of GDM became elevated with the increasing number of risk factors (P < 0.001). Early GCT was performed in 1687 (16.1%) pregnant women and the early detected GDMs only accounted for 11.9% of all GDMs among those with at least one risk factor. Among those who had early GCT, the GDM diagnosis rate increased with the number of risk factors (P < 0.001). Previous analysis in this survey identified 4 independent risk factors for GDM among 16 risk factors: BMI > or = 24, age over 30 years old, family history of DM and southerners. Similar analysis was performed according to the above 4 risk factors and similar results were found as those found for 16 risk factors. No significant difference was found in the GDM and GIGT incidence between the two analyses in those with at least one risk factor. CONCLUSION: Early GCT is necessary for pregnant women with risk factors of GDM, but the screening rate in China is low. GCT should be repeated for those women with risk factors of GDM and normal GCT at early screening.

[Effect of integrative Chinese and Western medicine in treating pregnant women with intrahepatic cholestasis].

OBJECTIVE: To evaluate the therapeutic effect of compound salvia injection combined with ursodeoxycholic acid (UDCA) in treating pregnant women with intrahepatic cholestasis (ICP) and its influence on perinatal babies. METHODS: One hundred and twenty-eight patients of ICP were assigned to two groups. The 72 patients in the treatment group were treated with salvia injection (20 mL in 10% glucose 500 mL for intravenous dripping once a day) and UDCA (15 mg, thrice daily by oral taken), and the 56 patients in the control group were treated with UDCA alone, all were treated for 14 days. Changes of itching symptom (estimated by scoring) and serum levels of biochemical indexes, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin (TBil) and glycocholic acid (GCA), were determined before and after treatment, and conditions of the newborns were compared after delivery. RESULTS: Compared with before treatment, scores of itching were lowered from 3.6 scores to 1.4 scores in the treatment group, and from 3.4 scores to 1.6 scores in the control group, showing no significant difference between groups (P > 0.05), but the lowering was shown earlier in the former. Levels of biochemical indexes were improved significantly (P < 0.01) in both groups, but the improvements were more significant in the treatment group, the difference between groups was significant (P < 0.05). The difference between groups in the incidence of fetal distress, meconium-stained fluid and neonatal asphyxia were insignificant (P > 0.05). The birth weights of the newborns were higher in the treatment group than in the control group (3,108 +/- 236 g vs 2,681 +/- 269 g, P < 0.05). CONCLUSION: The combined therapy of compound salvia injection and UDCA shows better effect in treating ICP than that of UDCA alone.

[Investigation on treatment of fetal growth restriction by salvia injection combined with composite amino acid].

OBJECTIVE: To explore the effect of salvia injection (SI) combined with composite amino acid (CAA) in treating fetal growth restriction (FGR). METHODS: A retrospective analysis was conducted on 106 pregnant women with FGR hospitalized from January 2007 to January 2008. Patients were randomized into 2 groups equally, the treated group (53 cases) treated with SI plus CAA, and the control group treated with CAA alone, all for 7 days. The clinical effect and umbilical blood flow (S/D) in patients were observed. RESULTS: (1) The total effective rate in the treated group was 81.13%, it was 88.80% (16/18) for patients in the gestation period of 24+ -28 weeks, 80.00% (12/15) for those of 28+ -32 weeks, and 75.00 (15/20) for 32+ -36 weeks, while in the control group, the corresponding rates were 69.81%, 77.77% (14/18), 66.66% (10/15), and 65.00% (13/20), respectively. The difference between the two groups was significant (P < 0.05). (2) After treatment, S/D significantly lowered in patients of the treated group (P < 0.05, P < 0.01), no matter how long the gestation period was, but it was insignificantly changed in the control group. CONCLUSION: The combined treatment with SI and CAA on FGR could improve the condition of the fetus.

[Impact of fetal growth restriction on spatial learning and memory abilities of the offspring of rats].

OBJECTIVE: To test the impact of fetal growth restriction (FGR) on the spatial learning and memory abilities of the offspring of rats. METHODS: FGR Model of Sprague-Dawley rats was constructed according to the method of passive smoking. The offspring of the rats were divided into male FGR group, male control group, female FGR group and female control group. Within each group, the rats were randomly divided into three subgroups to be tested at 1, 2, and 4 months of age, respectively (n =10 for each subgroup). Morris water maze task was performed to assess the spatial learning and memory abilities of the rats. RESULTS: The escape latencies to find the platform were shortened with increased training times for all of the rats. At the age of 1 and 2 months, both male and female rats in the FGR group spent more time in finding the platform than their counterparts in the control group (P < 0.05). At the age of 4 months, significant prolonged latency was only found in the female rats. The rats in the FGR group, except the 4 months old male rats, were more likely to use non-effective strategies (random or marginal strategies) to find the platform than the efficient strategies (tendency or straight strategies). The rats in the FGR group stayed in the platform shorter than those in the control groups (P < 0.05). CONCLUSION: FGR can cause gender- and age-specific impairment of spatial learning and memory abilities to the offspring.

Autoantibodies against glucose-regulated protein 78 as serological biomarkers in metastatic and recurrent hepatocellular carcinoma.

PURPOSE: To identify Heptocellular carcinoma (HCC) associated antigens by proteomics, and validate whether autoantibodies against tumor-associated antigens (TAAs) could be used for diagnosis and conditional monitoring. RESULTS: The 78 kDa glucose regulated protein (GRP78) was selected as a candidate TAA. The titers of autoantibodies against 78 kDa glucose regulated protein (GRP78) from patients with HCC, liver cirrhosis (LC), and chronic hepatitis (CH) were significantly higher than that from normal controls (P<0.05, P<0.001, and P<0.01, respectively). The expression of autoantibodies against GRP78 was associated with clinical stage (P<0.01), portal vein invasion (P<0.05), and metastasis (P<0.05). The expression of anti-GRP78 antibodies was significantly higher 1 month after surgery in recurrent patients who had accepted hepatic resection 1 month after surgery compared to patients who had surgery before surgery or within 1 week after surgery (P<0.01 and P<0.001). Immunohistochemistry (IHC) showed higher expression of GRP78 in HCC compared to the non-HCC liver tissues (P <0.05). MATERIALS AND METHODS: HCC serum with high titer of autoantibodies against TAAs were screened and used for a proteome-based approach to identify HCC associated antigens. Indirect enzyme-linked immunoassay (ELISA) was used to detect the corresponding autoantibodies against TAAs. CONCLUSION: GRP78 is an autoantigen that could stimulate autoimmune responses and serve as a potential marker for recurrent and metastatic progression in HCC.

FAM3A attenuates ER stress-induced mitochondrial dysfunction and apoptosis via CHOP-Wnt pathway.

Endoplasmic reticulum (ER) stress is linked to several neurological disorders, and neuronal injury cascades initiated by excessive ER stress are mediated, in part, via mitochondrial dysfunction. In the present study, we identified FAM3A as an important regulator of ER stress-induced cell death in neuronal HT22 cells. The ER stress inductor tunicamycin (TM) significantly decreased the expression of FAM3A at both mRNA and protein levels, which was shown to be dependent on the induction of reactive oxygen species (ROS). Overexpression of FAM3A attenuated TM-induced apoptosis and activation of ER stress factors, but had no effect on ER calcium metabolism in HT22 cells. We also found decreased mitochondrial ROS generation, inhibited cytochrome c release and preserved mitochondrial membrane potential (MMP) in FAM3A overexpressed cells. In addition, the experiments using isolated mitochondria showed that overexpression of FAM3A attenuated mitochondrial swelling and loss of mitochondrial Ca(2+) buffering capacity after TM exposure. By using specific targeted small interfering RNA (siRNA) to knockdown the expression of the C/EBP homologous protein (CHOP), we found that FAM3A-induced protection and inhibition of ER stress was mediated by inverting TM-induced decrease of Wnt through the CHOP pathway. Our study demonstrates a pivotal role of FAM3A in protecting against TM-induced cytotoxicity via regulating CHOP-Wnt pathway, and suggests the therapeutic values of FAM3A overexpression against ER stress-associated neuronal injury.

Clinical characteristics of fetal and neonatal outcomes in twin pregnancy with preeclampsia in a retrospective case-control study: A STROBE-compliant article.

The aim of our study was to compare the clinical characteristics of fetal and neonatal outcomes in twin pregnancies between women with preeclampsia (PE) and those with normotension in a Chinese population.There were 143 preeclamptic women and 367 normotensive women with twin pregnancies included in this retrospective case-control study. The baseline characteristics and perinatal outcomes were collected and compared between the groups. Multiple logistic regression and linear regression were used to assess the correlations between PE and the outcomes.Significant increases were observed in the frequencies of preterm delivery (OR = 2.75, P < 0.001), iatrogenic preterm birth (OR = 3.52, P < 0.001), and IUGR (OR = 2.94, P = 0.001) in the PE group, and the PE group had more than a 2-fold risk of adverse neonatal outcomes. Preeclamptic twin neonates had lower birth weights (ß = -147.34, P = 0.005; ß = -169.47, P = 0.001). The comparison on the discordance of intertwin weight was not significantly different.Twin pregnancies with PE are associated with worse perinatal outcomes. The adverse outcomes of preeclamptic twin pregnancies may be associated with lower birth weights rather than the discordance of the intertwin weight, which requires further confirmation. The results may provide helpful references for better clinical assessments, evaluations of prognosis, and a deeper understanding of preeclamptic twin pregnancies.

[Effect of behavioral training on learning and memory capacity and changes of hippocampal NR2B and GluR1 expressions in FGR offspring rats].

OBJECTIVE: To study the effect of behavioral training on the learning and memory abilities and changes of NR2B and GluR1 expressions in the hippocampus of offspring rats with fetal growth retardation (FGR). METHODS: A FGR model was established in SD rats by passive smoking. The offspring rats were divided into FGR group and control group, each then randomized into training and untrained group. Morris water maze behavioral training was carried out in postnatal months 2 and 4, and the learning and memory abilities of the young rats were assessed using dark-avoidance test and step-down test. NR2B and GluR1 expression in the hippocampus of the rats were detected by immunohistochemistry. RESULTS: In the dark-avoidance and step-down tests, the FGR rats showed deteriorated learning and memory performance in comparison with the control group, but behavioral training resulted in improved performance of the rats. The performance in FGR group was much improved after behavioral training, and the model factor and the training factor showed a significant interaction (P<0.05). The expression of NR2B and GluR1 in CA1 and CA3 regions of the hippocampus decreased in FGR group, then the their expressions in the CA1 region increased after training in both FGR and control groups, and the increment was especially obvious in GluR1 expression in the CA1 region at postnatal month 2. The two factors showed a significant interaction (P<0.05). CONCLUSION: Behavioral training can improve the learning and memory abilities of FGR offspring rats, the mechanism of which is probably related to increased expression of NR2B and GluR1 in the CA1 region of the hippocampus.

Difference between 2 h and 3 h 75 g glucose tolerance test in the diagnosis of gestational diabetes mellitus (GDM): results from a national survey on prevalence of GDM.

The possibility of the 2 h oral glucose tolerance test (OGTT) as an alternative to the 3 h OGTT was investigated based on data from a national survey on pregnancy-associated diabetes. Data were retrieved from 4179 pregnant women who had OGTT performed after an abnormal 50 g glucose challenge test (GCT). All of the 4 glucose levels during their OGTT were collected and analyzed. According to American Diabetes Association (ADA) gestational diabetes mellitus (GDM) diagnostic criteria, among the 4179 pregnant women who required OGTT, 3429 (82.1%) were normal and 750 (17.9%) were diagnosed as GDM. If the 3rd h glucose levels were omitted from OGTT, 79 cases of GDM (10.5%) would be overlooked. No trend was shown where women with more risk factors were more likely to be overlooked if the 3rd h test was omitted (χ2 for trend=0.038, P>0.05). No significant differences were found in the rate of cesarean section (CS), preterm births or macrosomia between the 79 cases and those with normal OGTT results and in the gestational weeks when OGTT was performed. It shows that in order to diagnose one woman with GDM, another 52 pregnant women would have an innocent 3rd h glucose test. Omission of the 3rd h glucose test in OGTT might be reasonable due to its convenience, better compliance and a small number of possibly miss-diagnosed cases, and their pregnancy outcomes have no significant difference from those of normal pregnant women.