MiR-186-5p Dysregulation Leads to Depression-like Behavior by De-repressing SERPINF1 in Hippocampus.

Diagnosis of major depressive disorder (MDD) is perplexing due to its multifactorial etiologies. Here, we isolated exosomes from the peripheral blood of MDD patients and healthy control subjects for mass spectrometry-based label-free quantitative proteomics. We identified that SERPINF1 is significantly diminished in the peripheral blood-derived exosomes of MDD patients compared to the healthy control subjects. Through RNA immunoprecipitation and luciferase reporter assays, we validated that SERPINF1 is a target of miR-186-5p that is upregulated in MDD patients' blood. In vivo studies in the chronic unpredictable mild stress (CUMS) mice further demonstrated that SERPINF1 in hippocampus is suppressed by miR-186-5p. Inhibiting the microRNA significantly restores the hippocampal SERPINF1 mRNA and protein expression, and ameliorates the depressive-like behaviors including sucrose preference and extended immobility time in the forced swim test. Instead, overexpressing miR-186-5p through tail intravenous injection of the mimics molecularly and behaviorally phenocopies the CUMS mice in wild-type mice. Our results indicate that the exosomal SERPINF1 in peripheral blood could serve as a reliable biomarker indicating MDD development, and miR-186-5p is a potential therapeutic target for the disease.

miR-30-5p-mediated ferroptosis of trophoblasts is implicated in the pathogenesis of preeclampsia.

Oxidative stress is a major cause of adverse outcomes in preeclampsia (PE). Ferroptosis, i.e. programmed cell death from iron-dependent lipid peroxidation, likely mediates PE pathogenesis. We evaluated specific markers for ferroptosis in normal and PE placental tissues, using in vitro (trophoblasts) and in vivo (rat) models. Increase in malondialdehyde content and total Fe2+ along with reduced the glutathione content and glutathione peroxidase activity was observed in PE placenta. While the trophoblasts experienced death under hypoxia, inhibitors of ferroptosis, apoptosis, autophagy, and necrosis increased the cell viability. Microarrays, bioinformatic analysis, and luciferase reporter assay revealed that upregulation of miR-30b-5p in PE models plays a pivotal role in ferroptosis, by downregulating Cys2/glutamate antiporter and PAX3 and decreasing ferroportin 1 (an iron exporter) expression, resulting in decreased GSH and increased labile Fe2+. Inhibition of miR-30b-5p expression and supplementation with ferroptosis inhibitors attenuated the PE symptoms in rat models, making miR-30b-5p a potential therapeutic target for PE.

Alpha-1-antitrypsin suppresses oxidative stress in preeclampsia by inhibiting the p38MAPK signaling pathway: An in vivo and in vitro study.

This present study was designed to investigate the effects of alpha-1-antitrypsin (AAT) on oxidative stress in preeclampsia (PE) by regulating p38 mitogen-activated protein kinase (p38MAPK) signaling pathway. HTR8/SVneo cells were randomly assigned into normal, hypoxia/reoxygenation (H/R), HR + AAT and HR + siRNA-AAT groups. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the mRNA and protein expressions of p-p38MAPK, AAT, signal transducer and activator of transcription 1 (STAT1) and activating transcription factor2 (ATF2). Flow cytometry, scratch test, cell counting kit-8 (CCK-8) assay and the 3-(4,5)-dimethylthiazol (-z-y1)-3,5-di- phenyltetrazolium bromide (MTT) assay were conducted to detect reactive oxygen species (ROS) and cell apoptosis, cell migration, proliferation and cytotoxicity, respectively. Mouse models in PE were established, which were divided into normal pregnancy (NP), PE and PE + AAT groups with blood pressure and urine protein measured. Chromatin immunoprecipitation (ChIP) and enzyme-linked immunosorbent assay (ELISA) were conducted to detect the activity of oxidative stress-related kinases and expressions of inflammatory cytokines and coagulation-related factors in cells and mice placenta. Immunohistochemistry, Western blotting and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were performed to detect AAT and p38MAPK expressions, apoptosis-related protein expressions, and apoptosis rate in mice placenta. Compared with the normal group, the H/R group had decreased expression of AAT, activity of superoxide dismutase (SOD) and GSH-Px, cell proliferation and migration, but increased p38MAPK, STAT1, ATF2, MDA, H2O2, inflammatory cytokines, coagulation-related factors, cell cytotoxicity, ROS, apoptotic factors and apoptosis rate. Compared with the H/R group, the HR + ATT group had increased expressions of AAT, activity of SOD and GSH-Px, cell proliferation and migration but decreased p38MAPK, STAT1, ATF2, malonyldialdehyde (MDA), H2O2, inflammatory cytokines and coagulation-related factors, cell cytotoxicity, ROS, apoptotic factors and apoptosis rate, while opposite results were observed in the HR + siRNA-ATT group. Compared with the NP group, the PE group had decreased activity of SOD and GSH-Px but increased MDA, H2O2, AAT, p38MAPK, inflammatory cytokines, coagulation-related factors and apoptosis rate. The indexes in the PE + AAT group were between the NP and PE groups. Thus, we concluded that AAT suppressed oxidative stress in PE by inhibiting p38MAPK signaling pathway.

Hysteroscopic treatment of postcesarean scar defect.

STUDY OBJECTIVE: To estimate the usefulness of hysteroscopy in the diagnosis and treatment of postcesarean scar defect. DESIGN: Retrospective study (Canadian Task Force classification III). SETTING: Two university-affiliated hospitals. PATIENTS: Sixty-two patients with postcesarean scar defects were retrospectively analyzed. INTERVENTIONS: All patients with postcesarean scar defects diagnosed using ultrasonography and hysteroscopy underwent hysteroscopic surgery, and were followed up for longer than 1 year. MEASUREMENTS AND MAIN RESULTS: Hysteroscopy revealed that 38 patients had valve-like motions at the incision sites, 22 had dome-like defects, and 2 with a history of 2 previous deliveries via cesarean section had umbilications of 2 different shades. Fifty-seven of 62 patients underwent corrective surgery via hysteroscopy. In another 3 patients, because the left wall of the fundus of the uterus was too thin (<2 mm at ultrasonography) to undergo corrective surgery, only clearance of residual blood and/or suture materials was performed. Of these 57 patients, 5 underwent removal of residual suture materials and endometrial fulguration. No complications were observed in these patients. Furthermore, after surgery, abnormal vaginal bleeding stopped in 38 patients, and its duration was shortened in 20 patients. In addition, dysmenorrhea was alleviated in 15 patients, and resolved in 7 patients. CONCLUSION: Hysteroscopy is an accurate means of diagnosis apart from surgical correction.

Alpha-1-antitrypsin acts as a preeclampsia-related protein: a proteomic study.

AIMS: To screen the preeclampsia-related protein by proteomics. METHODS: Proteomics was performed to identify differential protein expression profiles between normal full-term pregnancy, early-onset severe preeclampsia (ES-PE) or late-onset severe preeclampsia (LS-PE; n = 10 per group). Real-time quantitative PCR and immunohistochemistry were conducted to confirm the expression of α(1)-antitrypsin (α(1)-AT) in the decidual tissues of different subjects. ELISA was employed to detect the α(1)-AT content in the peripheral blood of 90 women (n = 30 per group). RESULTS: We successfully constructed two-dimensional electrophoresis maps of decidual tissues, and a total of 20 differentially expressed proteins were identified. The α(1)-AT expression was different among the three groups. The normal full-term pregnancy women expressed the most α(1)-AT, and the LS-PE women expressed the least amount of α(1)-AT. The difference in the α(1)-AT expression was consistent with the proteomics data. The peripheral α(1)-AT content was the highest in the normal full-term pregnancy group (1.85 ± 0.15 g/l), moderate in the ES-PE group (0.77 ± 0.14 g/l) and lowest in the LS-PE group (0.42 ± 0.07 g/l; p < 0.05). CONCLUSION: Using 2D PAGE, we identified twenty proteins with significantly altered expression in PE. These differentially expressed proteins include prevention protein, in which α(1)-AT is downregulated in PE.

[Expression of Rho-GDP dissociation inhibitor in the decidual tissues of preeclampsia patients and its clinical implication].

OBJECTIVE: To investigate the expression of Rho-GDI in the decidual tissues of patients preeclampsia and explore its clinical implication. METHODS: Real-time PCR, Western blotting and immunohistochemistry were used to detect the mRNA and protein expressions of Rho-GDI in the decidual tissues from 30 normal women with full-term pregnancy, 30 patients with early-onset severe preeclampsia and 30 with late-onset severe preeclampsia. RESULTS: Rho-GDI expression was found mainly on the cell membrane and in the cytoplasm and nuclei of the decidual cells, occasionally occurring in the stroma. Both the mRNA and protein expressions of Rho-GDI in the decidual tissues were significantly higher in the normal pregnancy group than in the two severe preeclampsia groups (P<0.05), and the patients with late-onset severe preeclampsia had the lowest expressions of Rho-GDI. CONCLUSION: The lowered expression of Rho-GDI in the deciduas might be involved in the pathogenesis and progression of preeclampsia.

[Expression of annexin V in decidua tissues of preeclampsia patients].

OBJECTIVE: To investigate the expression of annexin V in the decidua tissues of preeclampsia patients and explore its clinical significance. METHODS: Real-time PCR, Western blotting and immunohistochemistry were employed to detect the mRNA and protein expressions of annexin V in the deciduas from 35 normal pregnant women at full term, 38 early onset severe preeclampsia patients and 33 late onset severe preeclampsia patients. RESULTS: Annexin V was found on the cell membrane and in the cytoplasm of the decidual cells and stroma. Both the mRNA and protein of annexin V expressions in the decidua tissues were significantly different between normal pregnancy group and early or late onset severe preeclampsia group (P<0.05), being the highest in normal pregnancy group and the lowest in early onset severe preeclampsia group. CONCLUSION: The low expression of annexin V in the deciduas might participate in the hypercoagulability state in preeclampsia patients.

Vascular endothelial growth factor-C siRNA delivered via calcium carbonate nanoparticle effectively inhibits lymphangiogenesis and growth of colorectal cancer in vivo.

A nonviral gene carrier, calcium carbonate (CaCO(3))-nanoparticle, was evaluated for the efficient in vitro and in vivo delivery of siRNA-targeting vascular endothelial growth factor-C (VEGF-C). The chemically synthesized CaCO(3)-nanoparticle has a 58-nm diameter and a + 28.6-mV positive surface charge. It is capable of forming a CaCO(3)-nanoparticle-DNA complex and transferring DNA into targeted cells with high transfection efficiency, while effectively protecting the encapsulated DNA from degradation. Further, the CaCO(3)-nanoparticle-DNA complex has no obvious cytotoxicity for LoVo cells, while a liposome-DNA complex exhibited measurable cytotoxicity. LoVo cells transfected with a VEGF-C-targeted small interfering RNA (siRNA) via the CaCO(3)-nanoparticle exhibits significantly reduced VEGF-C expression, as measured by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay, whereas no decrease in VEGF-C expression is observed in cells treated by control transfection. Transfection of LoVo cells with VEGF-C siRNA via the CaCO(3)-nanoparticle also dramatically suppresses tumor lymphangiogenesis, tumor growth, and regional lymph-node metastasis in subcutaneous xenografts. Significant downregulation of VEGF-C messenger RNA expression in a subcutaneous xenograft derived from VEGF-C siRNA-treated LoVo cells was confirmed by real-time PCR, as compared to controls. We conclude that the CaCO(3)-nanoparticle is a novel, nonviral system for the effective delivery of siRNA for cancer gene therapy.