Exploring the shared molecular mechanism of microvascular and macrovascular complications in diabetes: Seeking the hub of circulatory system injury

BackgroundMicrovascular complications, such as diabetic retinopathy (DR) and diabetic nephropathy (DN), and macrovascular complications, referring to atherosclerosis (AS), are the main complications of diabetes. Blindness or fatal microvascular diseases are considered to be identified earlier than fatal macrovascular complications. Exploring the intrinsic relationship between microvascular and macrovascular complications and the hub of pathogenesis is of vital importance for prolonging the life span of patients with diabetes and improving the quality of life.Materials and methodsThe expression profiles of GSE28829, GSE30529, GSE146615 and GSE134998 were downloaded from the Gene Expression Omnibus database, which contained 29 atherosclerotic plaque samples, including 16 AS samples and 13 normal controls; 22 renal glomeruli and tubules samples from diabetes nephropathy including 12 DN samples and 10 normal controls; 73 lymphoblastoid cell line samples, including 52 DR samples and 21 normal controls. The microarray datasets were consolidated and DEGs were acquired and further analyzed by bioinformatics techniques including GSEA analysis, GO-KEGG functional clustering by R (version 4.0.5), PPI analysis by Cytoscape (version 3.8.2) and String database, miRNA analysis by Diana database, and hub genes analysis by Metascape database. The drug sensitivity of characteristic DEGs was analyzed.ResultA total of 3709, 4185 and 8086 DEGs were recognized in AS, DN, DR, respectively, with 1820, 1666, 888 upregulated and 1889, 2519, 7198 downregulated. GO and KEGG pathway analyses of DEGs and GSEA analysis of common differential genes demonstrated that these significant sites focused primarily on inflammation-oxidative stress and immune regulation pathways. PPI networks show the connection and regulation on top-250 significant sites of AS, DN, DR. MiRNA analysis explored the non-coding RNA upstream regulation network and significant pathway in AS, DN, DR. The joint analysis of multiple diseases shows the common influenced pathways of AS, DN, DR and explored the interaction between top-1000 DEGs at the same time.ConclusionIn the microvascular and macrovascular complications of diabetes, immune-mediated inflammatory response, chronic inflammation caused by endothelial cell activation and oxidative stress are the three links linking atherosclerosis, diabetes retinopathy and diabetes nephropathy together. Our study has clarified the intrinsic relationship and common tissue damage mechanism of microcirculation and circulatory system complications in diabetes, and explored the mechanism center of these two vascular complications. It has far-reaching clinical and social value for reducing the incidence of fatal events and early controlling the progress of disabling and fatal circulatory complications in diabetes

Knockdown of Mediator Complex Subunit 19 Suppresses the Growth and Invasion of Prostate Cancer Cells.

Prostate cancer (PCa) is one of the most common cancers in elderly men. Mediator Complex Subunit 19 (Med19) is overexpressed and plays promotional roles in many cancers. However, the roles of Med19 in PCa are still obscure. In this study, by using immunohistochemical staining, we found higher expression level of Med19 in PCa tissues than in adjacent benign prostate tissues. We then knocked down the Med19 expression in PCa cell lines LNCaP and PC3 by using lentivirus siRNA. Cell proliferation, anchor-independent growth, migration, and invasion were suppressed in Med19 knockdown PCa cells. In nude mice xenograft model, we found that Med19 knockdown PCa cells formed smaller tumors with lower proliferation index than did control cells. In the mechanism study, we found that Med19 could regulate genes involved in cell proliferation, cell cycle, and epithelial-mesenchymal transition, including P27, pAKT, pPI3K, IGF1R, E-Cadherin, N-Cadherin, Vimentin, ZEB2, Snail-1 and Snail-2. Targeting Med19 in PCa cells could inhibit the PCa growth and metastasis, and might be a therapeutic option for PCa in the future

Development of a Real-Time Recombinase-Aided Amplification Method to Rapidly Detect Methicillin-Resistant <i>Staphylococcus aureus</i>

Methicillin-resistant staphylococcus aureus (MRSA) is a major pathogen responsible for human hospital and community-onset diseases and severe invasive livestock infections. Rapid detection of MRSA is essential to control the spread of MRSA. Conventional identification methods and antibacterial susceptibility tests of MRSA are time-consuming. The commonly used qPCR assay also has the disadvantages of being complicated and expensive, restricting its application in resource-limited clinical laboratories. Here, a real-time fluorescent recombinase-assisted amplification (RAA) assay targeting the most conserved regions within the mecA gene of MRSA was developed and evaluated to detect MRSA. The detection limit of this assay was determined to be 10 copies/reaction of positive plasmids. The established RAA assay showed high specificity for MRSA detection without cross-reactivities with other clinically relevant bacteria. The diagnostic performance of real-time RAA was evaluated using 67 clinical S. aureus isolates from dairy farms, which were detected in parallel using the TaqMan probe qPCR assay. The results showed that 56 and 54 samples tested positive for MRSA by RAA and qPCR, respectively. The overall agreement between both assays was 97.01% (65/67), with a kappa value of 0.9517 (p R2 = 0.9012, p < 0.0001), indicating that this RAA assay possesses similar detection performance to the qPCR assay. In conclusion, our newly established RAA assay is a time-saving and convenient diagnostic tool suitable for MRSA detection and screening

Flexible Free-Standing Luminescent Two-Component Fiber Films with Tunable Hierarchical Structures Based on Hydrogen-Bonding Architecture

Although the fabrication of hierarchical architectures with highly ordered functional units is of great importance for both fundamental science and practical application, the development of one-dimensional (1D) organic hierarchical micro/nanostructures based on low-molecular-weight (LMW) building blocks remains at an early stage. Herein, we report two types of micro/nanoscaled multicomponent fluorescent fiber systems with tunable hierarchical morphologies through a one-step coassembly process. With the aid of hydrogen-bonding motifs, LMW precursors (1,4-bis­(5-phenyloxazol-2-yl)­benzene (<b>A</b>) and two coassembled building blocks: 4-bromotetrafluorobenzene carboxylic acid (<b>B</b>) and 2,3,4,5,6-pentafluorophenol (<b>C</b>)) have been self-organized into fibers and flexible free-standing films, which show hierarchical micro/nanostructures as well as tunable one-/two-photon luminescence. The disassembly of the multicomponent <b>A.B</b> and <b>A.C</b> fibers occurs at high temperature, which further alters the luminescence properties of the multicomponent materials. Therefore, this work provides a facile wet chemical route for fabricating multicomponent LMW self-assembled fibers and free-standing film systems with tunable hierarchical structures and photoemission behaviors, and such self-organized systems may have potential applications in fields of two-photon luminescence and thermal sensors

Plasmid-mediated tigecycline-resistant gene tet

ABSTRACTThe recent emergence of plasmid-mediated tigecycline resistance genes, tet(X3) and tet(X4), in animals and humans in China would pose a foreseeable threat to public health. To illustrate this paradigm shift in tigecycline resistance, here, covering the period 2008-2018, we retrospectively analysed a national strain collection of Escherichia coli (n = 2254), obtained from chickens and pigs, in six representative provinces of China. The gene tet(X4) was identified in five pig isolates collected in 2016 and 2018 from the provinces of Sichuan (3/15, 2018), Henan (1/25, 2018) and Guangdong (1/28, 2016), but not in the isolates prior to 2016. None of the isolates was detected harbouring tet(X3). All tet(X4)-positive E. coli exhibited high levels of tigecycline resistance (MICs, 16-64 mg/L), and two were confirmed as colistin resistant, harbouring chromosome-borne mcr-1 gene. The gene tet(X4) was detected on a plasmid in all five isolates, whereas a co-location of tet(X4) on the chromosome of one isolate was observed. Diverse host strains and novel plasmids related to the tet(X4) gene were observed. Our timely findings of the recent emergence of tet(X4) gene in food animal support the rapid surveillance and eradication of this gene before it is established

Establishment and characterization of LNCaP-Med19-si/sc and PC3-Med19-si/sc cells.

<p>(A) After lentivirus infection and FACS sorting, the transduction efficiency was confirmed by determining the GFP expression in a fluorescent microscope. The GFP positive rate was more than 95% in all cells. (B) In Q-PCR assay, the relative Med19 expression levels were 26.18±5.27% in LNCaP-Med19-si cells (n = 3, **<i>P</i><0.01), and 16.88±3.40% in PC3-Med19-si cells (n = 3, **<i>P</i><0.01). (C) In Western blot assay, the Med19 expression level was dramatically decreased in LNCaP-Med19-si and PC3-Med19-si cells. GAPDH was used as an internal control.</p

Knockdown of Med19 altered multiple genes expression in PCa cells.

<p>(A) In Western blot assay, the pAKT and pPI3K expression levels were lower, and the P27 expression level was higher in the Med19 knockdown cells. GAPDH was used as an internal control. (B, C) In Q-PCR assay, the IGF1R, N-Cadherin, Vimentin, ZEB2, Snail-1, and Snail-2 were down-regulated, and the E-Cadherin was up-regulated in both LNCaP-Med19-si and PC3-Med19-si cells (n = 3, *<i>P</i><0.05, **<i>P</i><0.01).</p

Knockdown of Med19 inhibited PCa cells invasion and migration <i>in vitro</i>.

<p>(A) In invasion assay, the invasion ratio was 4.26±0.92% in LNCaP-Med19-si cells and 11.74±1.94% in LNCaP-Med19-sc cells (n = 3, **<i>P</i><0.01). The invasion ratio was 19.71±4.00% in PC3-Med19-si cells and 32.27±3.72% in PC3-Med19-sc cells (n = 3, *<i>P</i><0.05). (B) In “wound healing” migration assay, after 24 hours culture, the percentage of wound healing area was 8.83±2.56% in LNCaP-Med19-si cells and 15.83±4.62% in LNCaP-Med19-sc cells (n = 3, **<i>P</i><0.01), 70.83±9.37% in PC3-Med19-si cells and 87.33±4.63% in PC3-Med19-sc cells (n = 3, **<i>P</i><0.01).</p