DRAIC mediates hnRNPA2B1 stability and m6A-modified IGF1R instability to inhibit tumor progression.

Type 1 insulin-like growth factor receptor (IGF1R) plays an important role in cancer, however, posttranscriptional regulation such as N6-methyladenosine (m6A) of IGF1R remains unclear. Here, we reveal a role for a lncRNA Downregulated RNA in Cancer (DRAIC) suppress tumor growth and metastasis in clear cell Renal Carcinoma (ccRCC). Mechanistically, DRAIC physically interacts with heterogeneous nuclear ribonucleoprotein A2B1 (hnRNPA2B1) and enhances its protein stability by blocking E3 ligase F-box protein 11 (FBXO11)-mediated ubiquitination and proteasome-dependent degradation. Subsequently, hnRNPA2B1 destabilizes m6A modified-IGF1R, leading to inhibition of ccRCC progression. Moreover, four m6A modification sites are identified to be responsible for the mRNA degradation of IGF1R. Collectively, our findings reveal that DRAIC/hnRNPA2B1 axis regulates IGF1R mRNA stability in an m6A-dependent manner and highlights an important mechanism of IGF1R fate. These findings shed light on DRAIC/hnRNPA2B1/FBXO11/IGF1R axis as potential therapeutic targets in ccRCC and build a link of molecular fate between m6A-modified RNA and ubiquitin-modified protein.

Eighteen iridoids from the roots and rhizomes of Valeriana jatamansi and their protective effects against α-hemolysin.

Thirteen previously undescribed iridoids (1-13), together with five known iridoids (14-18) were isolated from the roots and rhizomes of Valeriana jatamansi Jones. Their structures with absolute configurations were elucidated by analysis of MS, NMR, optical rotation and their experimental and calculated electronic circular dichroism spectra. All of the isolated compounds were tested for their protective effects against α-hemolysin-induced cell death in A549 cells. Compounds 14, 16 and 17 showed moderate protective effects, and compounds 15 and 18 showed weak protective effects.

Dimeric ent-kauranoids isolated from Isodon japonica var. Glaucocalyx and their anti-inflammatory activities.

The phytochemical investigation of the aerial parts of Isodon japonica var. glaucocalyx afforded four undescribed (glaucocalyxin O-R, 1-4) and six known ent-kauranoids (5-10). Their structures were established using NMR and MS measurements. Compounds 1 and 2 are dimeric ent-kaurane-type diterpenoids. Moreover, the plausible biogenetic pathways for compounds 1 and 2 were proposed as Michael addition between two monomers. Eight compounds were assayed for their anti-inflammatory activity by evaluating NO production in LPS-induced RAW 267.4 cells, and compounds 7, 8 and 9 exhibited relatively remarkable anti-inflammatory activities at 10 µM.

A Floating Capsule Electrochemical System for In Situ and Multichannel Ion-Selective Sensing.

Free-floating electrochemical sensors are promising for in situ bioprocess monitoring with the advantages of movability, a lowered risk of contamination, and a simplified structure of the bioreactor. Although floating sensors were developed for the measurement of physical and chemical indicators such as temperature, velocity of flow, pH, and dissolved oxygen, it is the lack of available electrochemical sensors for the determination of the inorganic ions in bioreactors that has a significant influence on cell culture. In this study, a capsule-shaped electrochemical system (iCapsuleEC) is developed to monitor ions including K+, NH4+, Na+, Ca2+, and Mg2+ based on solid-contact ion-selective electrodes (SC-ISEs). It consists of a disposable electrochemical sensor and signal-processing device with features including multichannel measurement, self-calibration, and wireless data transmission. The capacities of the iCapsuleEC were demonstrated not only for in situ measurement of ion concentrations but also for the optimization of the sensing electrodes. We also explored the possibility of the system for use in detection in simulated cell culture media.

Whole genome analysis of Bacillus amyloliquefaciens TA-1, a promising biocontrol agent against Cercospora arachidicola pathogen of early leaf spot in Arachis hypogaea L.

BACKGROUND: Early leaf spot disease, caused by Cercospora arachidicola, is a devastating peanut disease that has severely impacted peanut production and quality. Chemical fungicides pollute the environment; however, Bacillus bacteria can be used as an environmentally friendly alternative to chemical fungicides. To understand the novel bacterial strain and unravel its molecular mechanism, De novo whole-genome sequencing emerges as a rapid and efficient omics approach. RESULTS: In the current study, we identified an antagonistic strain, Bacillus amyloliquefaciens TA-1. In-vitro assay showed that the TA-1 strain was a strong antagonist against C. arachidicola, with an inhibition zone of 88.9 mm. In a greenhouse assay, results showed that the TA-1 strain had a significant biocontrol effect of 95% on peanut early leaf spot disease. De novo whole-genome sequencing analysis, shows that strain TA-1 has a single circular chromosome with 4172 protein-coding genes and a 45.91% guanine and cytosine (GC) content. Gene function was annotated using non-redundant proteins from the National Center for Biotechnology Information (NCBI), Swiss-Prot, the Kyoto Encyclopedia of Genes and Genomes (KEGG), clusters of orthologous groups of proteins, gene ontology, pathogen-host interactions, and carbohydrate-active enZYmes. antiSMASH analysis predicted that strain TA-1 can produce the secondary metabolites siderophore, tailcyclized peptide, myxochelin, bacillibactin, paenibactin, myxochelin, griseobactin, benarthin, tailcyclized, and samylocyclicin. CONCLUSION: The strain TA-1 had a significant biological control effect against peanut early leaf spot disease in-vitro and in greenhouse assays. Whole genome analysis revealed that, TA-1 strain belongs to B. amyloliquefaciens and could produce the antifungal secondary metabolites.

The Relevant Approaches for Aligning Carbon Nanotubes.

Carbon-nanotube (CNT) is a promising material owing to its compelling mechanical, thermal and electrical properties and has been applied in a broad variety of fields such as composite, fiber, film and microelectronic. Although the introductions of CNT have brought huge improvement for many applications, these properties of macrostructures prepared by CNTs still cannot meet those of individual CNT. Disordered alignment of CNTs in the matrix results in degradation of performance and hinders further application. Nowadays, quantities of methods are being researched to realize alignments of CNTs. In this paper, we introduce the application of CNTs and review some typical pathways for vertical and horizontal alignment, including chemical vapor disposition, vertical self-assembly, external force, film assisted, electric field, magnetic field and printing. Besides that, advantages and disadvantages of specific methods are also discussed. We believe that these efforts will contribute to further understanding the nature of aligned CNT and generating more effective ideas to the relevant workers.

Long non-coding RNA MALAT1 enhances angiogenesis during bone regeneration by regulating the miR-494/SP1 axis.

Bone regeneration is a coordinated process involving connections between blood vessels and osteocytes. Angiogenesis and osteogenesis are tightly connected throughout the progression of bone regeneration. This study aimed to explore the underlying mechanism of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1)-regulated angiogenesis during bone regeneration. Gene and protein expression was detected by quantitative real-time PCR and western blot assay. Vascular endothelial growth factor (VEGFA) secretion was assessed by enzyme-linked immunosorbent assay. To evaluate the effect of osteogenic differentiation, alkaline phosphatase (ALP) and alizarin red staining assays were performed. Proliferation was detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Migration and angiogenesis were measured using Transwell and tube formation assays. A dual luciferase reporter assay was performed to confirm the binding relationship among MALAT1, miR-494, and specificity protein 1 (SP1). Expression levels of MALAT1, SP1, and VEGFA were elevated and miR-494 was suppressed in MC3T3-E1 cells after culture in osteogenic medium. MALAT1 knockdown suppressed the osteogenic differentiation of MC3T3-E1, since ALP activity, mineralized nodules, and expression of the osteodifferentiated markers runt-related transcription factor 2 and osterix were restrained. In addition, MALAT1 silencing inhibited angiogenesis during bone regeneration, as the proliferation, migration, and capillary tube formation of human umbilical vein endothelial cells were blocked. Furthermore, miR-494 was directly targeted by MALAT1 and regulated the SP1/Toll-like receptor 2 (TLR2)/bone morphogenetic protein 2 (BMP2) axis by targeting SP1. Furthermore, miR-494 overexpression inhibited angiogenesis and osteogenic differentiation. Moreover, SP1 overexpression or miR-494 inhibition rescued the regulatory effect of sh-MALAT1 on angiogenesis and osteogenic differentiation. Taken together, these findings indicate that MALAT1 promotes angiogenesis and osteogenic differentiation by targeting miR-494 and activating the SP1/TLR2/BMP2 pathway, suggesting a novel target for bone regeneration therapy by promoting angiogenesis.

Clinical characteristics, treatment, and prognosis of 74 2019 novel coronavirus disease patients in Hefei: A single-center retrospective study.

ABSTRACT: Since December 2019, pneumonia caused by a novel coronavirus (SARS-CoV-2), namely 2019 novel coronavirus disease (COVID-19), has rapidly spread from Wuhan city to other cities across China. The present study was designed to describe the epidemiology, clinical characteristics, treatment, and prognosis of 74 hospitalized patients with COVID-19.Clinical data of 74 COVID-19 patients were collected to analyze the epidemiological, demographic, laboratory, radiological, and treatment data. Thirty-two patients were followed up and tested for the presence of the viral nucleic acid and by pulmonary computed tomography (CT) scan at 7 and 14 days after they were discharged.Among all COVID-19 patients, the median incubation period for patients and the median period from symptom onset to admission was all 6 days; the median length of hospitalization was 13 days. Fever symptoms were presented in 83.78% of the patients, and the second most common symptom was cough (74.32%), followed by fatigue and expectoration (27.03%). Inflammatory indicators, erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP) of the intensive care unit (ICU) patients were significantly higher than that of the non-ICU patients (P < .05). However, 50.00% of the ICU patients had their the ratio of T helper cells to cytotoxic T cells (CD4/CD8) ratio lower than 1.1, whose proportion is much higher than that in non-ICU patients (P < .01).Compared with patients in Wuhan, COVID-19 patients in Anhui Province seemed to have milder symptoms of infection, suggesting that there may be some regional differences in the transmission of SARS-CoV-2 between different cities.

The Long Noncoding RNA ANRIL Promotes Cell Apoptosis in Lipopolysaccharide-Induced Acute Kidney Injury Mediated by the TLR4/Nuclear Factor-Kappa B Pathway.

BACKGROUND/AIMS: The purpose of this study is to analyze the expression and biological function of lncRNA ANRIL, microRNA-199a, TLR4, and nuclear factor-kappa B (NF-κB) in acute renal injury (AKI) induced by lipopolysaccharide (LPS). METHODS: The levels of ANRIL and microRNA-199a in mouse cells and kidneys were detected by quantitative-polymerase chain reaction. Western blot analysis was used for the NF-κB pathway protein. MTT assay was used for cell viability. Enzyme-linked immunosorbent assay was used for the secretion of inflammatory factors in mouse kidney tissue. Apoptosis was measured by flow cytometry and Western blotting. The potential binding region between ANRIL and miR-199a was verified by luciferase reporter assay. RESULTS: The upregulation of ANRIL can reduce the expression of microRNA-199a and increases the number of apoptotic cells. The expression levels of ANRIL in LPS-induced AKI mice and LPS-treated HK2 cells were upregulated compared with the control group. Overexpression of ANRIL increased apoptosis and promoted TLR4 (Toll-like receptor 4), NF-κB phosphorylation, and downstream transcription factor production. CONCLUSION: ANRIL/NF-κB pathway in LPS-induced apoptosis provided theoretical guidance for ANRIL in the treatment of AKI.

SP1/TGF­ß1/SMAD2 pathway is involved in angiogenesis during osteogenesis.

The relationship between osteoblasts and angiogenesis is vital for bone regeneration, especially mandibular and maxillary bones. Transforming growth factor ß1 (TGF­ß1) and vascular endothelial growth factor (VEGF) are closely related to angiogenesis; however, the regulatory mechanism between them remains unknown. The present study aimed to reveal this mechanism to provide novel insight for development of potential therapeutic opportunities. Western blotting and reverse transcription­quantitative PCR was used to assess the protein and mRNA expression levels in MC3T3­E1 preosteoblast cells and HUVECs, ELISAs were used to detect the expression levels of secreted VEGF, MTT assays were used to assess the viability of the cells, migratory ability was assessed using Transwell assays, angiogenesis assays were used to analyze the formation of blood vessels, and TGF­ß1 regulation was confirmed using a dual­luciferase reporter assay. The overexpression of specificity protein 1 (SP1) or TGF­ß1 increased VEGF expression levels and secretion, and promoted angiogenesis of co­cultured HUVECs. SP1 also promoted SMAD2 phosphorylation. These effects of SP1 were all reversed by the TGF­ß1 inhibitor. The VEGF inhibitor bevacizumab also reduced the SP1/TGF­ß1/SMAD2 pathway­induced angiogenesis of preosteoblasts. In conclusion, it was demonstrated that SP1 promoted TGF­ß1 expression, activated the SMAD2 pathway and induced VEGF secretion, which may enhance angiogenic processes in preosteoblasts.

CYP2J2-produced epoxyeicosatrienoic acids attenuate ischemia/reperfusion-induced acute kidney injury by activating the SIRT1-FoxO3a pathway.

Cytochrome P450 (CYP) epoxygenases can metabolize arachidonic acids to epoxyeicosatrienoic acids (EETs), which play a protective role in the renal system, but their involvement in ischemia/reperfusion (I/R)-induced acute kidney injury remains unknown. Here, using a rat model, we demonstrated that forced CYP2J2 expression attenuated I/R-induced renal dysfunction and protected histological integrity. We showed that CYP2J2 significantly decreased I/R-induced upregulation of blood urea nitrogen and serum creatinine and enhanced autophagy during I/R treatment. In addition, we determined the protective effect of CYP2J2 against I/R-caused apoptosis. We demonstrated that CYP2J2 overexpression attenuated the downregulation of SIRT1 and FoxO3a by I/R-induced injury. Moreover, exogenous 11,12-EET addition obviously promoted I/R-induced autophagic flux and suppressed I/R-induced apoptosis through SIRT1-FoxO3a signaling activation. Our data indicate that CYP2J2-produced EETs improve I/R-caused kidney injury by activating the SIRT1-FoxO3a signaling pathway, which protects from renal I/R injury.

Olmesartan Medoxomil, An Angiotensin II-Receptor Blocker, Ameliorates Renal Injury In db/db Mice.

BACKGROUND: Diabetic nephropathy (DN) is a common complication of diabetes mellitus (DM) and also a major cause of end-stage renal disease (ESRD). Olmesartan medoxomil (OM) is an angiotensin II receptor blocker (ARB) and has been shown to exhibit renoprotective effects on a streptozotocin (STZ)-induced diabetic rat model. Yet, whether OM affects DN progression and renal injury in db/db mice, a type 2 diabetic murine model, has not been established. METHODS: Wild-type (n = 15) and db/db mice (n = 15) were treated with control saline or OM via oral gavage. The physiological and biochemical parameters were evaluated and histological examinations of kidney specimens were performed. RESULTS: Compared with saline-treated db/db mice, db/db mice administered with OM showed ameliorated diabetic physiological and biochemical parameters. In addition, OM decreased urinary albumin excretion and plasma creatinine level in db/db mice. Moreover, histologically, OM reduced glomerular hypertrophy and injury, and also ameliorated tubular injury, thus suggesting that OM improves renal function and minimizes renal pathological deterioration in db/db mice. CONCLUSION: Our study reveals a beneficial role of OM in ameliorating DN in db/db mice, which is associated with its renoprotective function.

Development and Validation of Dummies and Human Models Used in Crash Test.

The crash test dummy, an important tool for car crash safety tests, is of great significance to explore the injury biomechanics of the occupants and improve the safety performance of the vehicle. The article mainly consists of four parts: brief introduction of injury mechanism, early experiments for obtaining biomechanical response (animal tests, cadaver tests, and volunteer tests), and development and validation of mechanical dummies and computational models. This study finds that the current crash test dummies are generally designed based on European and American, so they have limitations on the damage prediction of other regions. Further research in the crash test dummy needs the participation of various countries in order to develop a crash test dummy that meets the national conditions of each country. Simultaneously, it is necessary to develop dummies of vulnerable groups, such as the elderly dummy and obese people dummy.

Biobjective Optimization Algorithms Using Neumann Series Expansion for Engineering Design.

In this paper, two novel algorithms are designed for solving biobjective optimization engineering problems. In order to obtain the optimal solutions of the biobjective optimization problems in a fast and accurate manner, the algorithms, which have combined Newton's method with Neumann series expansion as well as the weighted sum method, are applied to deal with two objectives, and the Pareto optimal front is achieved through adjusting weighted factors. Theoretical analysis and numerical examples demonstrate the validity and effectiveness of the proposed algorithms. Moreover, an effective biobjective optimization strategy, which is based upon the two algorithms and the surrogate model method, is developed for engineering problems. The effectiveness of the optimization strategy is proved by its application to the optimal design of the dummy head structure in the car crash experiments.

Thrombospondin-1 Production Regulates the Inflammatory Cytokine Secretion in THP-1 Cells Through NF-κB Signaling Pathway.

Thrombospondin-1 (TSP-1) is upregulated in several inflammatory diseases. Recent data have shown that macrophages from TSP-1-deficient mice have a reduced inflammatory phenotype, suggesting that TSP-1 plays a part in macrophage activation. DNA microarray approach revealed that Porphyromonas gingivalis lipopolysaccharide (P. gingivalis LPS) may induce the enhanced TSP-1 expression in human monocytes, suggesting a role of TSP-1-mediated pathogenesis in periodontitis. Until recently, the function of TSP-1 has been a matter of debate. In this study, we explored the role of TSP-1 in inflammatory cytokine secretions and its putative mechanism in pathogenesis of periodontitis. We demonstrated that TSP-1 expression was significantly upregulated in gingival tissues with periodontitis and in P. gingivalis LPS-stimulated THP-1 cells. Deficiency of TSP-1 by transfecting siRNAs decreased IL-6, IL-1ß, and TNF-α secretions in THP-1 cells, whereas overexpression of TSP-1 resulted in an upregulation of IL-6, IL-1ß, and TNF-α productions. Additional experiments showed that Pyrrolidine dithiocarbamate (PDTC) inhibited IL-6, IL-1ß, and TNF-α expression induced by overexpression of TSP-1, accompanying with downregulation of phosphorylated p65 and IκBα protein levels in response to P. gingivalis LPS. These results indicated that TSP-1 played a significant role in P. gingivalis LPS-initiated inflammatory cytokines (IL-6, IL-1ß, and TNF-α) secretions of THP-1 cells, and the NF-κB signaling is involved in its induction of expression. Thus, TSP-1 effectively elevated P. gingivalis LPS-induced inflammation mediated by the NF-κB pathway and may be critical for pathology of periodontitis.

Platelet-rich plasma promotes angiogenesis of prefabricated vascularized bone graft.

PURPOSE: The objective was to assess the impact of platelet-rich plasma (PRP) on angiogenesis and bone formation of tissue-engineered bone in the prefabricated stage. MATERIALS AND METHODS: Both thighs of New Zealand white rabbits were used as prefabricated vascularized bone grafts using a combination of bone mesenchymal stem cells and vascular bundles in a titanium cage filled with ß-tricalcium phosphate ceramic. PRP was applied in the test group, and the same procedure was performed in the control group without the application of PRP. After 4, 8, and 12 weeks, delayed static bone scanning with technetium-99m methylene diphosphonate was performed before sacrifice, and the tissue-engineered bone samples were collected for immunohistochemical analysis using a monoclonal antibody against CD31 and histologic analysis. RESULTS: The results showed superior angiogenesis in the PRP group compared with the control group at each time point as determined by bone scintigraphy and immunohistochemical examinations. The results of histologic analysis also showed that there was more bone formation in the PRP group than in the control group at each time point. CONCLUSIONS: The application of autologous PRP was an effective strategy for increasing angiogenesis and bone formation in tissue-engineered bone and had potential significance for clinical applications.

[Effects of Porphyromonas gingivalis on inflammatory factor secreted by human umbilical vein endothelial cells].

PURPOSE: To investigate the effect of porphyromonas gingivalis (P.g) on interleukin-18 (IL-18) and CRP secretion in human umbilical vein endothelial cells (HUVECs). METHODS: Anaeropack method was used for P.g cultivation and then HUVECs were co-cultured with P.g followed by assaying the amount of IL-18 and CRP with enzyme-linked immunosorbent assay (ELISA) kit. Linear regression and variance analysis were performed with SPSS11.0 software package. RESULTS: The unstimulated HUVECs expressed small amounts of IL-18 and CRP, P.g dose-dependently increased the production of IL-18 and CRP in HUVECs. Elevated IL-18 and CRP were found at 4-hour, then continued to increase at 8-hour, 12-hour and 24-hour after co-cultivation; under the effect of same concentration, the quantity of CRP was significantly higher than that of IL-18 secreted by HUVECs. There was significant different, P<0.05. CONCLUSION: It is concluded that P.g increased the production of IL-18 and CRP in HUVECs in a dose and time dependent manner, elevated IL-18 and CRP may be involved in the regulation of inflammatory both in periodontal disease and formation of atherosclerotic plaque in cardiovascular disease(CVD).