hsa-miR-15b-5p regulates the proliferation and apoptosis of human vascular smooth muscle cells by targeting the ACSS2/PTGS2 axis.

A previous bioinformatic analysis from our group predicted that the interaction of microRNA (miRNA/miR)-15b with the acyl-CoA synthetase short chain family member 2 (ACSS2) gene was important for the development of abdominal aortic aneurysm (AAA). Apoptosis of aortic vascular smooth muscle cells (VSMCs) is a pathological feature of AAA. The present study aimed to explain the roles of miR-15b/ACSS2 in AAA by exploring their effects on the proliferation and apoptosis of aortic VSMCs. Human aortic VSMCs (T/G HA-VSMC cell line) were divided into six groups and were transfected with miR-15b-5p mimics, mimic negative control (NC), miR-15b-5p inhibitors, inhibitor NC, miR-15b-5p mimics+pcDNA3.1 and miR-15b-5p mimics+ACSS2-overexpessing vector. CCK-8 assay was used to determine cell proliferation. Annexin V-FITC/PI staining and flow cytometry assays were used to measure cell apoptosis. Dual-luciferase reporter assays were used to confirm the targeted relationship between miR-15b-5p and ACSS2. Reverse transcription-quantitative PCR and/or western blotting were used to examine the expression levels of miR-15b-5p, ACSS2 and prostaglandin-endoperoxide synthase 2 (PTGS2). Following transfection of T/G HA-VSMCs with mimics and inhibitors to respectively upregulate and downregulate miR-15b-5p, the results demonstrated that overexpression of miR-15b-5p inhibited cell proliferation and promoted cell apoptosis; silencing of miR-15b-5p obtained the opposite results. ACSS2 may be a direct target of miR-15b-5p, since the luciferase activity of a ACSS2 wild-type vector, but not that of a ACSS2 mutant reporter, was significantly inhibited by miR-15b-5p mimics compared with controls. Additionally, the expression levels of ACSS2 and its downstream gene PTGS2 were significantly reduced or increased following transfection with miR-15b-5p mimics or inhibitors, respectively. Furthermore, overexpression of ACSS2 reversed the antiproliferative and proapoptotic effects of miR-15b-5p mimics by blocking the production of PTGS2 protein. In conclusion, miR-15b-5p may promote the apoptosis and inhibit the proliferation of aortic VSMCs via targeting the ACSS2/PTGS2 axis. The present study provided preliminary evidence indicating that the miR-15b-5p/ACSS2/PTGS2 axis may be a potential target for the treatment of AAA.

miRNAs regulating the expressions of NTF3, GNG2 and ITGA7 are involved in the pathogenesis of abdominal aortic aneurysm in mice.

Abdominal aortic aneurysm (AAA) is a life-threatening vascular disease, but effective treatment strategies remain lacking. The objective of this study was to screen underlying therapeutic targets by investigating the molecular mechanisms of AAA using mouse models. The mRNA (GSE109639) and miRNA (GSE51229 and GSE54943) expression profiles of mouse AAA models were downloaded from Gene Expression Omnibus database. A total of 1367 differentially expressed genes (DEGs) were identified between AAA and sham group, 490 of which were used for constructing the Protein-Protein Interaction (PPI) network. NTF3, GNG2 and ITGA7 in the PPI network were suggested to be hub genes according to their ranking of topological features. Furthermore, hub gene GNG2 was enriched in module 1, while ITGA7 was enriched in module 3. Eighteen differentially expressed miRNAs (DEMs) were shared in two datasets, 6 of which were predicted to regulate 130 DEGs (i.e. mmu-miR-677-ITGA7, mmu-miR-350-NTF3 and mmu-miR-292-3p-GNG2) to establish the miRNA-mRNA regulatory network. Function enrichment analysis showed NTF3 was involved in cell motion and MAPK signaling pathway; ITGA7 affected extracellular matrix (ECM)-receptor interaction; GNG2 participated in cell proliferation and chemokine signaling pathway. In conclusion, miRNAs regulating the expressions of NTF3, GNG2 and ITGA7 may represent underlying targets for treatment of AAA.

miR-30a-GNG2 and miR-15b-ACSS2 Interaction Pairs May Be Potentially Crucial for Development of Abdominal Aortic Aneurysm by Influencing Inflammation.

Abdominal aortic aneurysm (AAA) is a lethal vascular degenerative disease for the elderly, but current therapeutic options are limited. This study was to explore the molecular mechanisms of AAA to screen underlying treatment targets for AAA. The gene and microRNA (miRNA) expression profiles of human AAA were downloaded from Gene Expression Omnibus database under accession number GSE57691, GSE62179, and GSE63541. Differentially expressed genes (DEGs) and microRNAs (miRNAs; DEMs) were identified using the Linear Models for Microarray data method. Protein-protein interaction (PPI) network, module analysis, and miRNA-mRNA regulatory network analyses were performed to screen hub genes and miRNAs that regulated the hub genes. The Database for Annotation, Visualization and Integrated Discovery was used to predict the functions of genes. GEPIA and Tumor-miRNA-Pathway online software were used to validate the expressions of crucial DEMs and DEGs in other cancers, respectively. As a result, in the GSE57691 dataset, a total of 584 DEGs were found to be specific for AAA, 521 of which were used for constructing the PPI network. ACSS2 (acyl-CoA synthetase short-chain family member 2), GNG2 (G protein subunit gamma 2), and CXCL1 (C-X-C motif chemokine ligand 1) and CCR7 (C-C motif chemokine receptor 7) were believed to be hub genes by calculating their topological features in the PPI network. Upregulated GNG2 could interact with CXCL1 and CCR7 to involve in chemokine signaling pathway, while downregulated ACSS2 was associated with lipid biosynthetic process. In the miRNA-mRNA regulatory network, ACSS2 was found to be regulated by hsa-miR-15b; hsa-miR-30a could modulate the expression of GNG2. In line with our analysis in AAA, GNG2, ACSS2, hsa-miR-30a, and hsa-miR-15b were also confirmed to be significantly upregulated or downregulated in several cancer types. In conclusion, hsa-miR-30a-GNG2 and hsa-miR-15b-ACSS2 interaction pairs may represent novel mechanisms for explaining the pathogenesis of AAA. Targeted regulation of them may be potential strategies for treatment of AAA.

Identification of genes associated with the effect of inflammation on the neurotransmission of vascular smooth muscle cell.

Vascular smooth muscle cell (VSMC) accumulation and hypertrophy are common in vascular disorders, and inflammation has a crucial role in the development of these diseases. To investigate the effect of inflammation on the neurotransmission of VSMC, bioinformatic analysis was performed, following next generation sequencing. Genes of lipopolysaccharide (LPS)-treated A7r5 cells and phosphate-buffered saline (PBS)-treated A7r5 cells were sequenced via next generation sequencing, and each assay was repeated three times. Differentially expressed genes (DEGs) were obtained using the NOISeq package in R. Subsequently, their potential functions were predicted by functional and pathway enrichment analyses using the Database for Annotation, Visualization and Integrated Discovery online tool. Interaction relationships of the proteins enriched in pathways associated with neurological diseases, the proteins which had interaction relationships with adrenoceptor α 1D (ADRA1D) or calcium voltage-gated channel subunit α1 S (CACNA1S), separately, were obtained from STRING, and protein-protein interaction (PPI) networks were constructed using Cytoscape software. A total of 2,038 DEGs, including 1,094 upregulated and 944 downregulated genes in the LPS treatment group were identified when compared with the control group. Enrichment analyses showed that NADH:Ubiquinone Oxidoreductase Core Subunit V2 (NDUFV2) was involved in several neurological diseases, including oxidative phosphorylation, Alzheimer's disease, Parkinson's disease and Huntington's disease. Furthermore, NDUFV2 (degree, 20) had a higher degree in the PPI network for DEGs enriched in pathways associated with neurological diseases. In the PPI network for ADRA1D, CACNA1S and the DEGs interacting with them, prohibitin (PHB), oxytocin receptor (OXTR), collapsin response mediator protein 1 (CRMP1) and dihydropyrimidinase like 2 (DPYSL2) had interaction relationships with both ADRA1D and CACNA1S. To conclude, the present study revealed that NDUFV2, PHB, OXTR, CRMP1 and DPYSL2 may have key roles in the effect of inflammation on neurotransmission of VSMC.

Identification of potential gene targets in systemic vasculitis using DNA microarray analysis.

The present study aimed to identify the involvement of critical genes in systemic vasculitis, to gain an improved understanding of the molecular circuity and to investigate novel potential gene targets for systemic vasculitis treatment. The dual­color cDNA microarray data of GSE16945, consisting of peripheral mononuclear blood cell specimens from 13 patients with systemic vasculitis and 16 healthy controls, was downloaded from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) were screened in systemic vasculitis compared with controls using BRB ArrayTools, followed by the construction of a protein­protein interaction (PPI) network using the clusterProfiler package, and significant functional interaction (FI) module selection. Furthermore, transcriptional factors (TFs) among the identified DEGs were predicted and a transcriptional regulation network was constructed. A total of 173 up- and 93 downregulated genes were identified, which were mainly associated with immune response pathways. FBJ murine osteosarcoma viral oncogene homolog (FOS), ubiquitin B (UBB), signal transducer and activator of transcription 1 (STAT1) and MX dynamin­like GTPase 1 (MX1) were identified as hub proteins in the PPI network. Furthermore, UBB, FOS, and STAT1 were hub proteins in the three identified FI modules, respectively. In total, nine TFs were predicted among the DEGs. Of the DEGs that were predicted to be TFs, STAT1, v­maf avian musculoaponeurotic fibrosarcoma oncogene homolog B (MAFB) and tyrosine 3­monooxygenase/tryptophan 5­monooxygenase activation protein Z (YWHAZ), which interacted with each other, were identified to regulate further DEGs as target genes. Various genes, including FOS, UBB, MX1, STAT1, MAFB, and YWHAZ may be potential targets useful for the treatment of systemic vasculitis.

Efficacy of Endovascular Radiofrequency Ablation for Thromboangiitis Obliterans (Buerger's Disease).

BACKGROUND: This study aimed to evaluate the efficacy of endovascular radiofrequency ablation (RFA) for the treatment of thromboangiitis obliterans (TAO). METHODS: Total 30 males (median age: 46.00 years, interquartile range: 42.00-51.25 years) with unilateral TAO in the lower extremity underwent RFA were retrospectively enrolled from January 2013 and October 2013. The pre-operative and post-operative digital subtraction angiographic (DSA) images were recorded. Pain scores preoperatively and postoperatively were assessed according to the World Health Organization Pain Guideline. The values of ankle brachial index (ABI) at pre-operation, post-operation, 2 weeks and 2 years after surgery were all recorded and analyzed. Additionally, a 2-year follow up was performed by a computed tomographic angiography (CTA) image. RESULTS: The DSA images indicated that occlusion of femoral artery was improved after surgery. Moreover, there was no recurrence of TAO at 2 years of follow-up based on the CTA images. The pain score (P < 0.001) was significantly deceased after surgery. The values of ABI at postoperation, 2 weeks after surgery, and 2 years after surgery were all significantly higher than the preoperative ABI (P < 0.001). Furthermore, the values of ABI at 2 weeks after surgery and 2 years after surgery were all significantly higher than the postoperative ABI (P < 0.001). CONCLUSIONS: These results supported the application of endovascular RFA for treating TAO.

Combined subfascial endoscopic perforator surgery and endovenous laser treatment without impact on the great saphenous vein for management of lower-extremity varicose veins.

BACKGROUND: Conventional high ligation and stripping of the great saphenous vein (GSV) has a good curative effect but is highly traumatic with a considerable relapse rate. Subfascial endoscopic perforator surgery (SEPS) plus endovenous laser treatment (EVLT) could be applied as individual therapy. This study aimed to evaluate the feasibility of performing combined SEPS and EVLT without impacting GSV in the management of valvular insufficiency of the lower-limb venous perforators. METHODS: Placement of lower-limb venous perforator insufficiency was marked by ascending phlebography in 83 affected limbs from September 2010 to June 2011. After randomization, SEPS was performed on 41 limbs to address the insufficiency of the venous perforators under the deep fascia, in combination with EVLT to close the superficial varicose veins without impacting the GSV. The remaining 42 limbs were treated using traditional GSV phlebectomy as controls. RESULTS: Postoperatively, all varicose veins were resolved, with lightening of the pigmentation and healing of the ulcer. Within a follow-up period of 5 - 11 months, no symptoms had recurred. Compared with the control group, the operation time, the number of incisions sutured, and the in-hospital time decreased on average by 1.5 hours, 4.7, and 6.8 days, respectively (P < 0.01 in all cases). CONCLUSION: Combined SEPS and EVLT for treatment of valvular insufficiency of the lower-limb venous perforators offer the advantages of microtrauma and rapid cure.

A retrospective study comparing endovenous laser ablation and microwave ablation for great saphenous varicose veins.

BACKGROUND: Endo-venous laser or microwave ablation is a minimally invasive surgery for treating varicose veins of lower limbs. AIM: The aim of our study was to determine whether endovenous microwave ablation of the greater saphenous vein was associated with better effectiveness and less complications than the endovenous laser ablation. MATERIALS AND METHODS: From July 2008 to June 2011, 259 cases (306 limbs) of varicose veins were assigned to endovenous laser ablation (n=138, 163 limbs) or endovenous microwave ablation (n=121, 143 limbs). RESULTS: Through analysis there was no significant difference of the operating time, length of hospital stay and Aberdeen score in the two groups. The recanalization rate was statistically higher in the laser group than that in the microwave group. The ecchymosis complication was significantly lower in microwave ablation than that of laser ablation group. However, the skin burn and paralysis complications were significantly lower in the laser ablation than that of microwave ablation group. CONCLUSIONS: Endo-venous microwave ablation is an effective alternative to laser ablation for treatment of varicose veins, associated with higher occlusion rate and without serious complications.

Electrospun emodin polyvinylpyrrolidone blended nanofibrous membrane: a novel medicated biomaterial for drug delivery and accelerated wound healing.

In this work, blended nanofibrous membranes were prepared by an electrospinning technique with polyvinylpyrrolidone (PVP) K90 as the filament-forming polymer, and emodin, an extract of polygonum cuspidate known as a medicinal plant, as the treatment drug. Detailed analysis of the blended nanofibrous membrane by scanning electron microscopy, Differential scanning calorimetry and X-ray diffraction revealed that emodin was well distributed in the ultrafine fibers in the form of amorphous nanosolid dispersions. Results from attenuated total reflectance Fourier transform infrared spectra suggested that the main interactions between PVP and emodin might be mediated through hydrogen bonding. In vitro dissolution tests proved that the blended nanofibrous membrane produced more desired release kinetics of the entrapped drug (emodin) as compared to the pure drug. Furthermore, wound healing test and histological evaluation revealed that the emodin loaded nanofibrous membrane to be more effective as a healing accelerator thereby proving potential strategies to develop composite drug delivery system as well as promising materials for future therapeutic biomedical applications.