Use of Transcriptome Sequencing to Analyze the Effects of Different Doses of an Astragalus-Rhubarb-Saffron Mixture in Mice with Diabetic Kidney Disease.

Purpose: To investigate the therapeutic effect and underlying mechanism of a traditional Chinese medicine (TCM) mixture consisting of Astragalus, rhubarb, and saffron in a mouse model of diabetic kidney disease (DKD). Methods: Forty-eight db/db mice received no TCM (DKD model), low-dose TCM, medium-dose TCM, or high-dose TCM, and an additional 12 db/m mice received no TCM (normal control). Intragastric TCM or saline (controls) was administered daily for 24 weeks. Blood glucose, body weight, serum creatinine (SCr), blood urea nitrogen (BUN), blood lipids, and urinary microalbumin were measured every four weeks, and the urinary albumin excretion rate (UAER) was calculated. After 24 weeks, kidney tissues were collected for transcriptome sequencing, and the main functions of these genes were determined via functional enrichment analysis. Results: Compared with the DKD model group, the medium-dose and high-dose TCM groups had significantly decreased levels of SCr, BUN, total cholesterol, triglycerides, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and UAER (all p<0.05). We identified 42 genes that potentially functioned in this therapeutic response, and the greatest effect on gene expression was in the high-dose TCM group. We also performed functional enrichment analysis to explore the potential mechanisms of action of these different genes. Conclusion: A high-dose of the Astragalus-rhubarb-saffron TCM provided the best prevention of DKD. Analysis of the kidney transcriptome suggested that this TCM mixture may prevent DKD by altering immune responses and oxygen delivery by hemoglobin.

Co-differential genes between DKD and aging: implications for a diagnostic model of DKD.

Objective: Diabetic kidney disease (DKD) is a serious complication of diabetes mellitus (DM) that is closely related to aging. In this study, we found co-differential genes between DKD and aging and established a diagnostic model of DKD based on these genes. Methods: Differentially expressed genes (DEGs) in DKD were screened using GEO datasets. The intersection of the DEGs of DKD and aging-related genes revealed DKD and aging co-differential genes. Based on this, a genetic diagnostic model for DKD was constructed using LASSO regression. The characteristics of these genes were investigated using consensus clustering, WGCNA, functional enrichment, and immune cell infiltration. Finally, the expression of diagnostic model genes was analyzed using single-cell RNA sequencing (scRNA-seq) in DKD mice (model constructed by streptozotocin (STZ) injection and confirmed by tissue section staining). Results: First, there were 159 common differential genes between DKD and aging, 15 of which were significant. These co-differential genes were involved in stress, glucolipid metabolism, and immunological functions. Second, a genetic diagnostic model (including IGF1, CETP, PCK1, FOS, and HSPA1A) was developed based on these genes. Validation of these model genes in scRNA-seq data revealed statistically significant variations in FOS, HSPA1A, and PCK1 gene expression between the early DKD and control groups. Validation of these model genes in the kidneys of DKD mice revealed that Igf1, Fos, Pck1, and Hspa1a had lower expression in DKD mice, with Igf1 expression being statistically significant. Conclusion: Our findings suggest that DKD and aging co-differential genes are significant in DKD diagnosis, providing a theoretical basis for novel research directions on DKD.

Identification and Functional Mechanism Verification of Novel MicroRNAs Associated with the Fibrosis Progression in Chronic Kidney Disease.

Chronic kidney disease (CKD) is a serious threat to human health worldwide, and its incidence is increasing annually. A growing amount of information is emerging about the role of micoRNAs (miRNAs) in the regulation of renal fibrosis, which has aroused interest in the development of drugs that block pathogenic miRNAs or restore protective miRNAs levels. To clarify the role of miRNAs in CKD, we selected patients with significant renal fibrotic disease (diabetic nephropathy (DN) and focal segmental glomerulosclerosis (FSGS)) as the disease group, and patients with little or no renal fibrotic disease (minimal change disease (MCD) and renal carcinoma adjacent to normal kidney) as controls. Significantly differentially expressed miRNAs were obtained by human kidney tissue sequencing, subsequently verified in mice models of DN and FSGS, and subsequently inhibited or overexpressed in human renal tubular epithelial cells (HK-2) stimulated by high glucose (HG) and TGF-ß1 in vitro. Therefore, the mechanism of its action in renal fibrosis was further elaborated. Finally, the downstream target genes of the corresponding miRNAs were verified by bioinformatics analysis, qRT-PCR, western blot and double luciferase report analysis. Two novel miRNAs, hsa-miR-1470-3p (miR-1470) and hsa-miR-4483-3p (miR-4483), were detected by renal tissue sequencing in the disease group with significant renal fibrosis (DN and FSGS) and the control group with little or no renal fibrosis (MCD and normal renal tissue adjacent to renal carcinoma). Subsequent human renal tissue qRT-PCR verified that the expression of miR-1470 was significantly increased, while the expression of miR-4483 was markedly decreased in the disease group (p < 0.05). Moreover, in vivo DN and FSGS mice models, the expression levels of miR-1470 and miR-4483 were consistent with the results of human kidney tissue. In vitro, miR-4483 was suppressed, whereas miR-1470 was induced by treatment with TGF-ß1 or HG. Inhibition of miR-1470 or overexpression of miR-4483 promoted HG or TGF-ß1-induced fibrosis in HK-2 cells. Further study revealed that MMP-13 and TIMP1 were the target genes ofmiR-1470 and miR-4483, respectively. Our study identifies newly dysregulated miRNA profiles related to fibrosis kidneys. miR-1470 and miR-4483 are demonstrated to participate in kidney fibrosis by regulation of MMP-13, TIMP1 respectively. Our results may represent a promising research direction for renal disorders and help identify new biomarkers and therapeutic targets for CKD.

Exosome miR-30a-5p Regulates Glomerular Endothelial Cells' EndMT and Angiogenesis by Modulating Notch1/VEGF Signaling Pathway.

BACKGROUND: Diabetic nephropathy (DN) is one of the microvascular complications of diabetes. Endothelial-mesenchymal transition (EndMT) and endothelial damage lead to abnormal angiogenesis in DN. OBJECTIVES: This study aimed to investigate the role of exosome miR-30a-5p in high glucose (HG)-induced glomerular endothelial cells (GECs) dysfunction and explore the underlying mechanisms. METHODS: GECs were cultured in normal glucose (5.5 mM) and HG (30 mM) conditions. The recipient GECs were transfected with exosome or miR-30a-5p mimic/inhibitor and then detected by using CCK-8 and flow cytometry assay. Luciferase analysis was used to verify miR-30a-5p acted on notch homolog protein 1 (Notch1). RT-qPCR and Western blot were used to detect the expression of VE-cadherin, α-SMA, vascular endothelial growth factor (VEGF) and Notch1. In vivo, exosome miR-30a-5p was administered to DN mice, and periodic acid-Schiff (PAS) staining, UTP levels, and HbA1c levels were measured. RESULTS: The expression of miR-30a-5p was downregulated in HG-treated GECs. Exosome miR-30a-5p significantly promoted cell proliferation, and migration and reduced apoptosis of GECs under HG conditions. MiR-30a-5p directly targeted the 3-UTR region of Notch1. Exosome miR-30a-5p reduced the expression levels of Notch1 and VEGF, both at mRNA and protein levels. Furthermore, exosome miR-30a-5p inhibited HG-induced EndMT, as evidenced by increased VE-cadherin and reduced α-SMA. In vivo studies demonstrated that exosome miR-30a-5p reduced serum HbA1c levels and 24-hour urine protein quantification. CONCLUSION: This study provides evidence that exosome miR-30a-5p suppresses EndMT and abnormal angiogenesis of GECs by modulating the Notch1/VEGF signaling pathway. These findings suggest that exosome miR-30a-5p could be a potential therapeutic strategy for the treatment of DN.

A review on the relationship between Arachidonic acid 15-Lipoxygenase (ALOX15) and diabetes mellitus.

Arachidonic acid 15-lipoxygenase (ALOX15), as one of the lipoxygenase family, is mainly responsible for catalyzing the oxidation of various fatty acids to produce a variety of lipid components, contributing to the pathophysiological processes of various immune and inflammatory diseases. Studies have shown that ALOX15 and its related products are widely distributed in human tissues and related to multiple diseases such as liver, cardiovascular, cerebrovascular diseases, diabetes mellitus and other diseases. Diabetes mellitus (DM), the disease studied in this article, is a metabolic disease characterized by a chronic increase in blood glucose levels, which is significantly related to inflammation, oxidative stress, ferroptosis and other mechanisms, and it has a high incidence in the population, accompanied by a variety of complications. Figuring out how ALOX15 is involved in DM is critical to understanding its role in diseases. Therefore, ALOX15 inhibitors or combination therapy containing inhibitors may deliver a novel research direction for the treatment of DM and its complications. This article aims to review the biological effect and the possible function of ALOX15 in the pathogenesis of DM.

A New Integrated GDL with Wavy Channel and Tunneled Rib for High Power Density PEMFC at Low Back Pressure and Wide Humidity.

Proton exchange membrane fuel cells (PEMFCs) have garnered significant attention due to their high efficiency and low emissions. However, PEMFC always suffers mass transfer and water management in performance improvement. Herein, an integrated gas diffusion layer (GDL) with wavy channel and micro-tunneled rib is designed and prepared to achieve faster and gentler mass transfer and excellent water management capability by laser engraving. Outstandingly, the new integrated GDL can use the back pressure of air as low as 0 and 50 kPa to respectively achieve 80% and 90% of fuel cell performance realized under pure oxygen. Such high performance is mainly due to the turbulent flow caused by wavy channel and express removing pathway of liquid water provided by micro-tunneled rib. Moreover, the new integrated GDL also shows wide humidity tolerance from 40% to 100% and a very high specific volume power density of 16,300 W L-1 due to the thin thickness of new integrated GDL. This new integrated GDL is expected to be widely used in PEMFC and other energy conversion devices.

Ordered Membrane Electrode Assembly with Drastically Enhanced Proton and Mass Transport for Proton Exchange Membrane Water Electrolysis.

In this work, an ordered membrane electrode assembly (MEA) based on a cone Nafion array with gradient Nafion distribution, tightly bonded catalytic layer/proton exchange membrane (CL/PEM) interface, and abundant vertical channels has been engineered by an anodic aluminum oxide template and magnetron sputtering method. Benefiting from a highly efficient CL/PEM interface, plentiful proton transfer highways, and rapid oxygen bubble release, this ordered MEA achieves an ultralow Ir loading of 20.0 µg cm-2 and a high electrochemical active area by 8.7 times compared to traditional MEA with Ir loading of 1.0 mg cm-2. It yields a mass activity of 168 000 mA mgIr-1 cm-2 at 2.0 V, which is superior to most reported PEM electrolyzers. Notably, this ordered MEA maintains excellent durability at a current density of 500 mA cm-2. This work opens a simple, cost-effective, and scalable route to design ordered MEAs for proton exchange membrane water electrolysis.

[Two new isoquinoline alkaloids from Corydalis hendersonii].

Corydalis hendersonii(CH) is a Tibetan folk medicine with the functions of clearing heat, detoxifying, cooling blood, checking diarrhea, and lowering blood pressure. It is often used to treat high altitude polycythemia, vasculitis, peptic ulcer, and diarrhea. Nine compounds were separated from the ethanol extract of CH by silica gel, ODS, Sephadex LH-20 chromatography and semi-preparative HPLC. Their structures were identified as hendersine H(1),hendersine I(2), dehydrocheilanthifoline(3), protopine(4), izmirine(5), 6,7-methylenedioxy-1(2H)-isoquinolinone(6), icariside D_2(7), ethyl 4-(ß-D-glucopyranosyloxy)-3-methoxybenzoate(8), 3-hydroxy-4-methoxybenzoic acid(9), respectively, by the spectroscopic data analysis and comparison with those in the literature. Among them, compounds 1 and 2 are new isoquinoline alkaloids, and compounds 7-9 are reported the first time for Corydalis. The hypoglycemic model of H9c2 cardiomyocytes and the inflammatory model of H9c2 cardiomyocytes induced by conditional supernatant were employed to determine the activities of the above compounds. The results showed that 20 µmol·L~(-1) compound 1 had a protective effect on H9c2 cardiomyocytes and 10 µmol·L~(-1) compounds 4 and 5 inhibited H9c2 cardiomyocyte inflammation induced by conditional supernatant.

Multiscale Architectured Nafion Membrane Derived from Lotus Leaf for Fuel Cell Applications.

Hierarchically patterned proton-exchange membranes (PEMs) have the potential to significantly increase the specific surface area, thus improving the catalyst utilization rate and performance of proton-exchange membrane fuel cells (PEMFCs). In this study, we are inspired by the unique hierarchical structure of the lotus leaf and proposed a simple three-step strategy to prepare a multiscale structured PEM. Using the multilevel structure of the natural lotus leaf as the original template, and after structural imprinting, hot-pressing, and plasma-etching steps, we successfully constructed a multiscale structured PEM with a microscale pillar-like structure and a nanoscale needle-like structure. When applied in a fuel cell, the multiscale structured PEM resulted in a 1.96-fold increase in discharge performance and a significant improvement in mass transfer compared to the membrane electrode assembly (MEA) with a flat PEM. The multiscale structured PEM has the combined advantage of a nanoscale and a microscale structure, benefiting from the markedly reduced thickness, increased surface area, and improved water management inherited from the multiscale structured lotus leaf's superhydrophobic characteristic. Using a lotus leaf as a multilevel structure template avoids the complex and time-consuming preparation process required by commonly used multilevel structure templates. Moreover, the remarkable architecture of biological materials can inspire novel and innovative applications in many fields through nature's wisdom.

Nanosized Proton Conductor Array with High Specific Surface Area Improves Fuel Cell Performance at Low Pt Loading.

The use of ordered catalyst layers, based on micro-/nanostructured arrays such as the ordered Nafion array, has demonstrated great potential in reducing catalyst loading and improving fuel cell performance. However, the size (diameter) of the basic unit of the most existing ordered Nafion arrays, such as Nafion pillar or cone, is typically limited to micron or submicron sizes. Such small sizes only provide a limited number of proton transfer channels and a small specific area for catalyst loading. In this work, the ordered Nafion array with a pillar diameter of only 40 nm (D40) was successfully prepared through optimization of the Nafion solvent, thermal annealing temperature, and stripping mode from the anode alumina oxide (AAO) template. The density of D40 is 2.7 × 1010 pillars/cm2, providing an abundance of proton transfer channels. Additionally, D40 has a specific area of up to 51.5 cm2/cm2, which offers a large area for catalyst loading. This, in turn, results in the interface between the catalyst layer and gas diffusion layer becoming closer. Consequently, the peak power densities of the fuel cells are 1.47 (array as anode) and 1.29 W/cm2 (array as cathode), which are 3.3 and 2.9 times of that without array, respectively. The catalyst loading is significantly reduced to 17.6 (array as anode) and 61.0 µg/cm2 (array as cathode). Thus, the nanosized Nafion array has been proven to have high fuel cell performance with low Pt catalyst loading. Moreover, this study also provides guidance for the design of a catalyst layer for water electrolysis and electrosynthesis.

A Recyclable Standalone Microporous Layer with Interpenetrating Network for Sustainable Fuel Cells.

The commercialization of fuel cells inevitably brings recycling problems. Therefore, achieving high recyclability of fuel cells is particularly important for their sustainable development. In this work, a recyclable standalone microporous layer (standalone MPL) with interpenetrating network that can significantly enhance the recyclability and sustainability of fuel cells is prepared. The interpenetrating network enables the standalone MPL to have high strength (17.7 MPa), gas permeability (1.55 × 10-13  m2 ), and fuel-cell performance (peak power density 1.35 W cm-2 ), providing the basic guarantee for its application in high-performance and highly recyclable fuel cells. Additionally, the standalone MPL is highly adaptable to various gas-diffusion backings (GDBs), providing high possibility to select highly recyclable GDBs. Outstandingly, anode standalone MPLs and GDBs can be easily detached from the spent membrane electrode assembly (MEA). This not only saves >90 vol% solvent in the recovery of the catalyst-coated membrane (CCM), but also extends the service life of the GDBs and the anode standalone MPL at least 138 times (2 760 000 h assuming 20 000 h of CCM) comparing to CCM. Therefore, the standalone MPL significantly enhances the recyclability and sustainability of fuel cells and is promising to be an indispensable component in the next-generation fuel cells.

The alkaloids of Corydalis hendersonii Hemsl. contribute to the cardioprotective effect against ischemic injury in mice by attenuating cardiomyocyte apoptosis via p38 MAPK signaling pathway.

BACKGROUND: There is a characteristic Tibetan folk medicine in China named Corydalis hendersonii Hemsl. (CH) has been used for treatment of cardiovascular related diseases, called "plethora" in Tibetan medicine. Previous studies demonstrated that ethanol extract of CH showed anti-acute myocardial infarction (AMI) effect through inhibiting fibrosis and inflammation. Rich alkaloids fraction (RAF) is isolated from CH, but whether RAF possessing an equivalent effect with the CH ethanol extract and by which mechanism it protects against AMI has not yet reported. The paper aimed to study the potential role of RAF on myocardial injured mice and its underlying mechanism. MATERIALS AND METHODS: Liquid chromatography mass spectrometry-ion trap-time of flight (LCMS-IT-TOF) was used to analyze the chemical profile and isolate pure compounds. The ligation of left anterior descending (LAD) of coronary artery in mice was used to evaluate the in vivo anti-AMI effect, by dividing into eight groups: Sham, Model, Fosinopril (10 mg/kg, i.g.), total extract (TE, 400 mg/kg, i.g.), poor alkaloids fraction  (PAF, 300 mg/kg, i.g.), and RAF (25, 50, and 100 mg/kg, respectively, i.g.) groups. Echocardiography was used to evaluate mice heart function through the index of left ventricular end-systolic  diameter (LVEDs), left ventricular end-diastolic diameter (LVEDd), fractional shortening (FS) and ejection fraction (EF). We detected the lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) in the serum and the plasma level of angiotensin II (AngII). The apoptosis of mice myocardial tissue was verified by TUNEL assay. The expression of p38 mitogen-activated protein kinases (p38 MAPK), Bcl-2 and Bcl-2-associated X protein (Bax) were detected through immunofluorescence staining, qRT-PCR and western blot in mice heart tissue and H9c2 cells. RESULTS: Echocardiography data indicated that the values of LVEDd and LVEDs were reduced and the values of FS and EF were improved by TE and RAF significantly. RAF also decreased the levels of LDH, CK-MB and AngII and significantly inhibited inflammatory cells in the marginal zone of myocardial infarction. The TUNEL assay results showed that RAF significantly attenuated cell apoptosis. Immunofluorescence and qRT-PCR assay showed that RAF inhibited p38 MAPK, Bax, and Bcl-2 proteins in mice myocardium. Western blot results validated that the expressions of key proteins were inhibited by RAF. Also, the apoptotic cells and apoptosis-related proteins were dramatically reduced by RAF in vivo and in vitro. Besides, RAF and PAF were analyzed by LCMS-IT-TOF to identify the main compounds and to demonstrate the difference between them. The results showed that a total of 14 alkaloids were identified, which indicated that the isoquinoline alkaloids were the main ingredients in RAF may contributing to the cardioprotective effect in mice. CONCLUSIONS: RAF improves cardiac function by inhibiting apoptosis via p38 MAPK signaling pathway, and RAF contributes to the effect against myocardial ischemic injury of TE in mice, which provides a substantial reference for the clinical application against ischemia heart disease and quality control of CH.

Integrated Gas Diffusion Electrode with High Conductivity Obtained by Skin Electroplating for High Specific Power Density Fuel Cell.

Smaller volume/weight and higher output power/energy density are always the goals of electrochemistry energy devices. Here, a simple strategy is proposed to prepare an integrated gas diffusion electrode (GDE) with high conductivity through skin electroplating. The skin electroplating is the combination of magnetron sputtering and spatial confinement electroplating. The electroplated metal obtained by skin electroplating is uniformly, continuously, and tightly attached to the surface of carbon paper like a layer of skin. Uniform and continuous electroplating metal layer endows the integrated electrode excellent conductivity with the square resistance as low as 27 mΩ sq-1 . In application, the self-breathing fuel cell with 1 cm2 active area can harvest ultrahigh volume specific power density (20.9 kW L-1 ). Additionally, the weight of the fuel cell stack (23 W) with the integrated electrode is only 20 g, which is only 7% of the commercial stack with the same power. The mass specific power density reaches 1150 W kg-1 , which is 15 times of the commercial stack. Outstandingly, the stack can charge 4 mobile phones at the same time. More importantly, the skin electroplating provides an effective strategy to improve the specific power density of other energy devices including Zn-air batteries, Li-air batteries, and so on.

Low-Coordination Trimetallic PtFeCo Nanosaws for Practical Fuel Cells.

Developing high-performance catalysts for fuel cell catalysis is the most critical and challenging step for the commercialization of fuel cell technology. Here 1D trimetallic platinum-iron-cobalt nanosaws (Pt3 FeCo NSs) with low-coordination features are designed as efficient bifunctional electrocatalysts for practical fuel cell catalysis. The oxygen reduction reaction (ORR) activity of Pt3 FeCo NSs (10.62 mA cm-2 and 4.66 A mg-1 Pt at 0.90 V) is more than 25-folds higher than that of the commercial Pt/C, even after 30 000 voltage cycles. Density functional theory calculations reveal that the strong inter-d-orbital electron transfer minimizes the ORR barrier with higher selectivity at robust valence states. The volcano correlation between the intrinsic structure featured with low-coordination Pt-sites and corresponding electronic activities is discovered, which guarantees high ORR activities. The Pt3 FeCo NSs located in the membrane electrode assembly (MEA) also achieve very high peak power density (1800.6 mW cm-2 ) and competitive specific/mass activities (1.79 mA cm-2 and 0.79 A mg-1 Pt at 0.90 ViR-free cell voltage) as well as a long-term lifetime in specific H2 O2 medium for proton-exchange-membrane fuel cells, ranking top electrocatalysts reported to date for MEA. This work represents a class of multimetallic Pt-based nanocatalysts for practical fuel cells and beyond.

Evaluation of malnutrition and inflammation after total parathyroidectomy in patients on maintenance dialysis.

PURPOSE: To evaluate the effect of total parathyroidectomy (tPTx) on malnutrition and inflammation in patients on maintenance dialysis (MHD) having secondary hyperparathyroidism (SHPT). METHODS: Twenty-five patients on MHD having SHPT who were being treated with tPTx were selected, and changes in their general condition (dry body mass), parathyroid hormone (PTH) and calcium levels, nutrition state (hemoglobin, hematocrit, serum albumin, and total iron binding capacity), and inflammatory status [serum C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor (TNF-α), and the malnutrition-inflammation score (MIS)] were observed at 12, 24, and 36 months postoperatively. RESULTS: Compared with the preoperative period, the dry body mass increased at 12, 24, and 36 months postoperatively (P < 0.01), hemoglobin, hematocrit, and serum albumin increased significantly (P < 0.01), whereas calcium, phosphorus, and PTH levels decreased significantly (P < 0.01). Serum CRP, IL-6, and TNF-α levels were significantly decreased at 12, 24, and 36 months after surgery (P < 0.01). Furthermore, MIS was reduced as well but to a lesser extent (P < 0.01). CONCLUSION: tPTx effectively reduced MIS in maintenance dialysis patients, and the alleviated malnutrition and improved inflammatory status may contributed to improving the quality of life of patients on MHD with SHPT.

Bioinformatics prediction and experimental verification of key biomarkers for diabetic kidney disease based on transcriptome sequencing in mice.

Background: Diabetic kidney disease (DKD) is the leading cause of death in people with type 2 diabetes mellitus (T2DM). The main objective of this study is to find the potential biomarkers for DKD. Materials and Methods: Two datasets (GSE86300 and GSE184836) retrieved from Gene Expression Omnibus (GEO) database were used, combined with our RNA sequencing (RNA-seq) results of DKD mice (C57 BLKS-32w db/db) and non-diabetic (db/m) mice for further analysis. After processing the expression matrix of the three sets of data using R software "Limma", differential expression analysis was performed. The significantly differentially expressed genes (DEGs) (-logFC- > 1, p-value < 0.05) were visualized by heatmaps and volcano plots respectively. Next, the co-expression genes expressed in the three groups of DEGs were obtained by constructing a Venn diagram. In addition, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were further analyzed the related functions and enrichment pathways of these co-expression genes. Then, qRT-PCR was used to verify the expression levels of co-expression genes in the kidney of DKD and control mice. Finally, protein-protein interaction network (PPI), GO, KEGG analysis and Pearson correlation test were performed on the experimentally validated genes, in order to clarify the possible mechanism of them in DKD. Results: Our RNA-seq results identified a total of 125 DEGs, including 59 up-regulated and 66 down-regulated DEGs. At the same time, 183 up-regulated and 153 down-regulated DEGs were obtained in GEO database GSE86300, and 76 up-regulated and 117 down-regulated DEGs were obtained in GSE184836. Venn diagram showed that 13 co-expression DEGs among the three groups of DEGs. GO analysis showed that biological processes (BP) were mainly enriched inresponse to stilbenoid, response to fatty acid, response to nutrient, positive regulation of macrophage derived foam cell differentiation, triglyceride metabolic process. KEGG pathway analysis showed that the three major enriched pathways were cholesterol metabolism, drug metabolism-cytochrome P450, PPAR signaling pathway. After qRT-PCR validation, we obtained 11 genes that were significant differentially expressed in the kidney tissues of DKD mice compared with control mice. (The mRNA expression levels of Aacs, Cpe, Cd36, Slc22a7, Slc1a4, Lpl, Cyp7b1, Akr1c14 and Apoh were declined, whereas Abcc4 and Gsta2 were elevated). Conclusion: Our study, based on RNA-seq results, GEO databases and qRT-PCR, identified 11 significant dysregulated DEGs, which play an important role in lipid metabolism and the PPAR signaling pathway, which provide novel targets for diagnosis and treatment of DKD.

Proteomics-based evaluation of the mechanism underlying vascular injury via DNA interstrand crosslinks, glutathione perturbation, mitogen-activated protein kinase, and Wnt and ErbB signaling pathways induced by crotonaldehyde.

Crotonaldehyde (CRA)-one of the major environmental pollutants from tobacco smoke and industrial pollution-is associated with vascular injury (VI). We used proteomics to systematically characterize the presently unclear molecular mechanism of VI and to identify new related targets or signaling pathways after exposure to CRA. Cell survival assays were used to assess DNA damage, whereas oxidative stress was determined using colorimetric assays and by quantitative fluorescence study; additionally, cyclooxygenase-2, mitogen-activated protein kinase pathways, Wnt3a, ß-catenin, phospho-ErbB2, and phospho-ErbB4 were assessed using ELISA. Proteins were quantitated via tandem mass tag-based liquid chromatography-mass spectrometry and bioinformatics analyses, and 34 differentially expressed proteins were confirmed using parallel reaction monitoring, which were defined as new indicators related to the mechanism underlying DNA damage; glutathione perturbation; mitogen-activated protein kinase; and the Wnt and ErbB signaling pathways in VI based on Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and protein-protein interaction network analyses. Parallel reaction monitoring confirmed significant (p < 0.05) upregulation (> 1.5-fold change) of 23 proteins and downregulation (< 0.667-fold change) of 11. The mechanisms of DNA interstrand crosslinks; glutathione perturbation; mitogen-activated protein kinase; cyclooxygenase-2; and the Wnt and ErbB signaling pathways may contribute to VI through their roles in DNA damage, oxidative stress, inflammation, vascular dysfunction, endothelial dysfunction, vascular remodeling, coagulation cascade, and the newly determined signaling pathways. Moreover, the Wnt and ErbB signaling pathways were identified as new disease pathways involved in VI. Taken together, the elucidated underlying mechanisms may help broaden existing understanding of the molecular mechanisms of VI induced by CRA.

Ultralight, Safe, Economical, and Portable Oxygen Generators with Low Energy Consumption Prepared by Air-Breathing Electrochemical Extraction.

Pure oxygen is vital in medical treatment, first aid, and chemical synthesis. Hypoxia can cause severe damage to the organ systems such as respiratory, digestive, and nervous systems and even directly cause death. Notably, the severe Coronavirus disease 2019 (COVID-19) pandemic has exacerbated the shortage of medical oxygen in the world. Hence, a safe, economical, and portable oxygen supply device is urgently needed. Here, we have successfully prepared a device with air-breathing electrochemical extraction of pure oxygen (ABEEPO) with light weight and high energy efficiency. By renovating the structure of the electrolytic cell, the components bipolar plate and end plate are replaced with a plastic membrane, and the component current collector is replaced with a highly conductive graphene composite membrane electrode. Due to the use of the plastic membrane and graphene composite membrane electrode, the weight of the electrolytic cell is reduced from 1319.4 to 1.6 g, and the flexibility of the electrolytic cell is successfully realized. Through optimizing anode catalysts, working area, and operating voltage, a high flow rate per mass (234 mL h-1 g-1) was achieved at a voltage of 1.2 V. The device exhibits high stability in 2 h. The new portable oxygen production device would be effective for hypoxia treatment.

The role of microRNA-155 in glomerular endothelial cell injury induced by high glucose.

OBJECTIVE: To investigate the role of microRNA-155-5p on apoptosis and inflammatory response in human renal glomerular endothelial cells (HRGEC) cultured with high glucose. METHODS: The primary HRGEC were mainly studied, light microscopy was used to detect changes in cell morphology. Quantitative Real Time-Polymerase Chain Reaction, Western Blot, immunofluorescence were aimed to observe the mRNA and protein expression levels of target gene ETS-1, downstream factors VCAM-1, MCP-1 and cleaved caspase-3 in each group after high glucose treatment as well as transfection with miR-155 mimics or inhibitor. RESULTS: The expression of inflammatory factors and apoptosis of HRGEC cells increased under high glucose treatment. Compared with normal-glucose treatment, the expression of microRNA-155 markedly increased in HRGECs treated with high-glucose, as well as the mRNA and protein levels of ETS-1, VCAM-1, MCP-1 and cleaved caspase-3. Overexpression of microRNA-155 remarkably downregulated mRNA and protein levels of ETS-1, VCAM-1, MCP-1 and cleaved caspase-3, whereas miRNA-155 knockdown upregulated their levels. In addition, HRGEC cells were transfected with miR-155 mimics and ETS-1 siRNA with high glucose stimulation. The expression of ETS-1 was positively correlated with the expression of downstream factors VCAM-1 and MCP-1. These results suggest that ETS-1 can mediate endothelial cell inflammation by regulating VCAM-1 and MCP-1. CONCLUSION: MiR-155 can negatively regulate the expression of target gene ETS-1 and its downstream factors VCAM-1, MCP-1 and cleaved caspase-3, thus mediating the inflammatory response and apoptosis of HRGEC.

Alashanoids O-S, seco-Humulane and Eremophilane Sesquiterpenoids from Syringa pinnatifolia.

Five new sesquiterpenoids, alashanoids O-S (1-5), along with three known analogs (6-8) were isolated from the peeled stems of Syringa pinnatifolia. Their structures were elucidated by analysis of extensive spectroscopic data including ESI-MS, 1D, 2D NMR. The absolute configurations were determined by comparing its experimental and calculated electronic circular dichroism, calculated OR, calculated NMR, and single crystal X-ray diffraction data analysis. Compounds 1 and 2 belong to the seco-humulane type and possess a rare 13-membered oxygen heterocycle framework, and 3-5 belong to eremophilane-type. Compounds 1, 2, and 5 showed inhibitory effects against NO production in LPS-induced RAW264.7 macrophage cells with its IC50 values of 11.86±2.34, 72.08±7.72, and 69.22±15.29 µM, respectively, compared with the positive control indomethacin (IC50 =31.52 µM).