Long non-coding RNA myocardial infarction associated transcript expression and clinical significance in patients with hypertension during pregnancy.

OBJECTIVE: This study aimed to investigate the expression level of lncRNA myocardial infarction associated transcript (MIAT) in serum of pregnant women with hypertensive disorders in pregnancy (HDP) and its clinical significance. METHODS: A total of 135 pregnant women with HDP were selected, including 69 pregnant women with gestational hypertension (GH) and 66 pregnant women with preeclampsia (PE). Sixty-eight normal pregnant women were selected as healthy control group (HC). The expression level of serum MIAT of all subjects was detected by quantitative real-time polymerase chain reaction (qRT-PCR), and the diagnostic value of MIAT for GH was evaluated by constructing receiver operating characteristic (ROC) curve. Pearson correlation coefficient was used to analyze the correlation between MIAT and patients' clinical indicators. Logistics regression analysis evaluated the influencing factors of GH development into PE. RESULTS: The level of MIAT in GH group was significantly higher than that in HC group, while MIAT level in PE group was more significantly upregulated than that in GH group and HC group. ROC curve showed that MIAT had the ability to distinguish between GH patients and healthy controls. Pearson correlation coefficient suggested that HOTAIR expression was positively correlated with systolic blood pressure (SBP) and diastolic blood pressure (DPB). Logistic regression analysis showed that MIAT was an independent influencing factor for the development of GH to PE. CONCLUSION: The expression of MIAT in serum of HDP patients was increased and positively correlated with the severity of the disease. The abnormal expression of MIAT has certain diagnostic value for GH.

Long Non-Coding RNA HOTAIR Expression and Clinical Significance in Patients with Gestational Diabetes.

PURPOSE: The global incidence of gestational diabetes mellitus (GDM) is increasing year by year, and many studies have proved that long non-coding RNA (lncRNA) is involved in the regulation of GDM. The purpose of this study was to investigate the expression of HOTAIR in GDM patients and its clinical significance. PATIENTS AND METHODS: Ninety-eight healthy pregnant women and 99 pregnant women diagnosed with GDM were enrolled in this study. Blood samples were collected from all participants and used for qRT-PCR analysis to determine the serum HOTAIR levels. The ROC curve was constructed to evaluate the diagnostic value of HOTAIR for GDM. Pearson correlation coefficient was used to estimate the correlation between HOTAIR and clinical indicators of patients. Logistic regression analysis was performed to evaluate the independent predictors of GDM. RESULTS: The level of HOTAIR was augmented in GDM group compared with healthy controls. ROC curve revealed that HOTAIR as a diagnostic marker of GDM has high sensitivity and specificity. Pearson correlation coefficient showed that HOTAIR level was positively correlated with body mass index, fasting plasma glucose, 1-hour plasma glucose and 2-hour plasma glucose. Logistic regression analysis shows that HOTAIR is an independent factor of the occurrence of GDM. CONCLUSION: The abnormal expression of HOTAIR in pregnant women with GDM made it a potential diagnostic biomarker for GDM.

111In- or 99mTc-labeled recombinant VEGF bioconjugates: in vitro evaluation of their cytotoxicity on porcine aortic endothelial cells overexpressing Flt-1 receptors.

INTRODUCTION: The aims of this study were to (a) synthesize and characterize a novel vascular endothelial growth factor (VEGF-2K) recombinant protein expressed in Pichia pastoris and (b) compare its cytotoxicity when labeled with the Auger electron emitter (111)In or (99m)Tc, both of which are in the nanometer-micrometer range, toward porcine aortic endothelial (PAE) cells transfected with the flt-1 gene to overexpress Flt-1 receptors (PAE-Flt-1). METHODS: The gene for the VEGF(165) isoform was fused to a sequence encoding an extended flexible peptide (KGGGGSK) with two accessible lysines for preferential derivatization with diethylenetriaminepentaacetic acid (DTPA) for complexing (111)In and a sequence for a His(6) affinity tag that bound the [(99m)Tc(CO)(3)(H(2)O)(3)](+) tricarbonyl complex. P. pastoris strain KM71H was transfected with the recombinant gene, the VEGF-2K protein expressed with methanol induction, and then purified by metal-affinity chromatography. VEGF-2K was modified with 13-mer peptides [CGYGPKKKRKVGG] containing the nuclear localization sequence (NLS) of SV-40 large T-antigen (underlined) to promote nuclear uptake following its receptor-mediated internalization. RESULTS: (99m)Tc-DTPA-VEGF-2K bound strongly and preferentially to PAE-Flt-1 cells compared with non-transfected PAE cells, but NLS modification diminished the ratio of PAE-Flt-1 to PAE binding to 2.3-fold. Nuclear accumulation of (99m)Tc-labeled DTPA-VEGF-2K was not enhanced by NLS modification but was enhanced by 1.5-fold for (111)In-DTPA-VEGF-2K-NLS. However, confocal microscopy revealed intranuclear distribution of DTPA-VEGF-2K-NLS, whereas DTPA-VEGF-2K distribution was mainly perinuclear. (111)In-DTPA-VEGF-2K-NLS was the most cytotoxic to PAE-Flt-1 cells, reducing their clonogenic survival by 4-fold. (111)In-DTPA-VEGF-2K, (99m)Tc-DTPA-VEGF-2K or (99m)Tc-DTPA-VEGF-2K-NLS had less effect on the clonogenic survival of PAE-Flt-1 or PAE cells. The strong cytotoxicity of (111)In-DTPA-VEGF-2K-NLS toward PAE-Flt-1 cells was associated with a 27-fold increase in nuclear foci of immunofluorescence for phosphorylated histone-2AX corresponding to sites of unrepaired DNA double-strand breaks. Monte Carlo modeling revealed that radionuclide decay in the nucleus would provide a 5-fold higher radiation absorbed dose for (111)In than for (99m)Tc, explaining their differential cytotoxicity, and intranuclear localization would amplify the radiation dose delivered by (111)In by 3-fold, explaining the greater potency of (111)In-DTPA-VEGF-2K-NLS compared with (111)In-DTPA-VEGF-2K. CONCLUSIONS: We conclude that targeted Auger electron radiotherapy aimed at Flt-1 receptors is a promising strategy that should be explored further for treatment of tumors in which this angiogenic pathway is up-regulated. (111)In is a more cytotoxic radionuclide than (99m)Tc, unless DNA delivery can be achieved, due to the short range of the electrons emitted.