Rapid and In-Field Sensing of Hydrogen Peroxide in Plant by Hydrogel Microneedle Patch.

The rapidly changing climate is exacerbating the environmental stress that negatively impacts crop health and yield. Timely sensing of plant response to stress is beneficial to timely adjust planting conditions, promoting the healthy growth of plants, and improving plant productivity. Hydrogen peroxide (H2O2) is an important molecule of signal transduction in plants. However, the common methods for detecting H2O2  in plants are associated with certain drawbacks, such as long extraction time, cumbersome steps, dependence on large instruments, and difficulty in realizing in-field sensing. Therefore, it is urgent to establish more efficient detection methods to realize the rapid detection of H2O2 content in plants. In this research, poly (methyl vinyl ether-alt-maleic acid) (PMVE/MA) hydrogel microneedle (MN) patch for rapid extraction of leaf sap are prepared, and the extraction mechanism of PEG-crosslinked PMVE/MA hydrogel MN patch is studied. A method of rapid detection of H2O2 content in plants based on MN patch with optical detection technology is constructed. The hydrogel MN patch can be used for timely H2O2 analysis. This application enables new opportunities in plant engineering, and can be extended to the safety and health monitoring of other plants and animals.

Pachymic acid activates TP53INP2/TRAF6/caspase-8 pathway to promote apoptosis in renal cell carcinoma cells.

While pachymic acid (PA), a key component of Poria cocos (Schw.), has demonstrated anti-tumor effects in lung, breast, and pancreatic cancers, its impact on renal cell carcinoma (RCC) is unclear. This study evaluated the effect of PA on proliferation, migration, and apoptosis in human renal cancer A498 and ACHN cells as well as in cancer xenograft mice using wound scratch test, Western blotting, and co-immunoprecipitation assays. In a dose- and time-dependent manner, PA exhibited significant inhibition of RCC cell proliferation, migration, and invasion, accompanied by the induction of apoptosis. Additionally, PA upregulated the expression of tumor protein p53-inducible nuclear protein 2 (TP53INP2) and tumor necrosis factor receptor-associated factor 6 (TRAF6), which were downregulated in renal papillary and chromophobe carcinoma, resulting in inhibited tumor growth in mice. PA treatment elevated cleaved-caspase 3 and 8, and PARP levels, and facilitated TP53INP2 and TRAF6 binding to caspase 8, promoting its ubiquitination. Molecular docking revealed interactions between PA and TP53INP2, TRAF6. In summary, PA inhibits RCC development by upregulating TP53INP2 and promoting TRAF6-induced caspase 8 ubiquitination, activating apoptotic pathways.

A Water-Driven and Low-Damping Triboelectric Nanogenerator Based on Agricultural Debris for Smart Agriculture.

The rapid progress in distributed electronics in agriculture depends on a wide range of energy supplies, such as cables and batteries. However, cable installation and maintenance are inconvenient in the agricultural environment, and the massive use of batteries will cause high replacement costs and serious environmental issues. To mitigate these problems, a water flow-driven and high-performance triboelectric nanogenerator based on agricultural debris (including derelict plant fibers and recycled greenhouse film) (AD-TENG) is developed. The precisely designed air gap and plant fiber-based dielectric brushes enable minimized frictional resistance and sustainable triboelectric charges, resulting in low damping and high performance for the AD-TENG. After nano-morphology modifications of the dielectric layer, the maximum power density of the AD-TENG increases by 64 times and reaches ≈1.24 W m-2 . The practical application demonstrates that the AD-TENG realizes the recycling of agricultural debris to achieve harvesting low-frequency and low-speed water-flow energy. Besides, the AD-TENG can be used to power agricultural sensors and develop the automatic irrigation system, which alleviates the energy consumption problem of agriculture and contributes to the realization of automated and informative intelligent agriculture.

Fluorinated Graphene-Enabled Durable Triboelectric Coating for Water Energy Harvesting.

The durability issue of triboelectric nanogenerators (TENGs) still limits its practical application for long-term operation, especially in harvesting water energy, which is one of the most widespread energies in nature. Attempting to ameliorate this, fluorinated graphene (FG) with unique triboelectric negativity and superhydrophobic property is introduced to serve as a new triboelectric and protective layer for TENG construction. The self-cleaning as well as anti-corrosion property and durability of the FG coating-enabled TENG device is then demonstrated. Furthermore, this novel triboelectronegative material candidate is applied for consecutively coating on Cu mesh and the inner wall of sewage pipe to fabricate TENGs for water energy harvesting and working as a metal protective layer in the meantime. Through the extraordinary output performance, the feasibility of FG-enabled triboelectric coating to be used in the field of interference screening, filtering, and sewage pipes is proved. This developed triboelectronegative coating not only broadens the material selection of TENG but also significantly enhances the durability and robustness of TENG for water energy harvesting, which possesses great potential in applying in metal protection and water energy harvesting under extreme conditions (e.g., strong acid and strong alkaline environments).

Growth-Controllable Triboelectric Nanogenerator Based on Surface-Attached Metal-Organic Framework Layer on Living Leaf.

Plant nanoelectronics aims to achieve specific functions by selecting suitable nanomaterials to connect or implant into plants. In this work, a new cyclic-spraying method is developed to attain controllable growth of surface-attached metal-organic framework (SURMOF) coatings on various complex substrates, including cotton, silk, and plant leaves. The covalent bonds formed between the SURMOF layer and substrate enable them to connect firmly without additional adhesive and do not fall off from the microstructured substrate surface with the change of biological activity or environment. Noteworthily, the triboelectric polarity of SURMOF can be regulated by changing the ligand molecule. As a proof of concept, a growth-controllable triboelectric nanogenerator (GC-TENG) based on living leaves and coated SURMOF layer is developed, and the feasibility of using it in the self-driven agricultural monitoring system is explored. In addition, long-term monitoring results show that the growth of SURMOF coating will not cause damage to plant leaf tissue, nor will it affect plant photosynthesis.

PINK1/Parkin mediated mitophagy ameliorates palmitic acid-induced apoptosis through reducing mitochondrial ROS production in podocytes.

Diabetic nephropathy (DN), the primary cause of end-stage renal disease (ESRD), is often accompanied by dyslipidemia, which is closely related to the occurrence and development of DN and even the progression to ESRD. Mitophagy, the selective degradation of damaged and dysfunctional mitochondria by autophagy, is a crucial mitochondrial quality control mechanism, and largely regulated by PINK1 (PTEN-induced putative kinase 1)/Parkin signaling pathway. In the present study, we demonstrated that PA induced mitochondrial damage and excessive mitoROS generation in podocytes. We also found PA treatment resulted in the activation of mitophagy by increasing co-localization of GFP-LC3 with mitochondria and enhancing the formation of mitophagosome, stabilization of PINK1 and mitochondrial translocation of Parkin, which indicated that PINK1/Parkin pathway was involved in PA-induced mitophagy in podocytes. Furthermore, inhibition of mitophagy by silencing Parkin dramatically aggravated PA-induced mitochondrial dysfunction, mitoROS production, and further enhanced PA-induced apoptosis of podocytes. Finally, we showed that PINK1/Parkin pathway were up-regulated in kidney of high fat diet (HFD)-induced obese rats. Taken together, our results suggest that PINK1/Parkin mediated mitophagy plays a protective role in PA-induced podocytes apoptosis through reducing mitochondrial ROS production and that enhancing mitophagy provides a potential therapeutic strategy for kidney diseases with hyperlipidemia, such as DN.

Alchemy-Inspired Green Paper for Spontaneous Recovery of Noble Metals.

Recycling of noble metal from waste materials, namely from electronic wastes (e-waste), spent catalyst, and industrial wastewater, is attracting growing attention due to the scarcity, economic importance, and criticality of those noble metals. Traditional techniques reported to date require toxic reagent and strict extraction conditions, which deeply hinders the development of precious metal recovery in complex environments. Here, an approach is proposed that uses flexible metallic transition metal dichalcogenide (TMD) paper, which provides abundant active sites for spontaneous adsorption and reduction of noble metal ions, as an Alchemy-inspired template to recover noble metal in an efficient and green way without the aid of reductant and heating. The metallic TMD (MoS2 , WS2 ) paper is shown to rapidly extract five noble metal ions (Au, Pd, Pt, Ag, and Ru) from complex samples containing various interferents. This unique property endows the metallic TMD paper with gifted ability in extracting gold from e-waste, and recovering platinum group metals (palladium and platinum) from spent catalysts, which provides a blueprint for the design of next-generation green platforms for noble metal regeneration.

Detection of Voltage Anomalies in Spacecraft Storage Batteries Based on a Deep Belief Network.

For a spacecraft, its power system is vital to its normal operation and capacity to complete flight missions. The storage battery is an essential component of a power system. As a spacecraft spends more time in orbit and its storage battery undergoes charge/discharge cycles, the performance of its storage battery will gradually decline, resulting in abnormal multivariate correlations between the various parameters of the storage battery system. When these anomalies reach a certain level, battery failure will occur. Therefore, the detection of spacecraft storage battery anomalies in a timely and accurate fashion is of great importance to the in-orbit operation, maintenance and management of a spacecraft. Thus, in this study, based on storage battery-related telemetry parameter data (including charge/discharge currents, voltages, temperatures and times) downloaded from an in-orbit satellite, a voltage anomaly detection algorithm for spacecraft storage batteries based on a deep belief network (DBN) is proposed. By establishing a neural network (NN) model depicting the correlations between each of the variables of temperature, current, pressure and charge/discharge times and voltage, this algorithm supports the detection of anomalies in the state-of-health of a storage battery in a timely fashion. The proposed algorithm is subsequently applied to the storage battery of the aforementioned in-orbit satellite. The results show the following. The anomalies detected using the proposed algorithm are more reliable, effective and visual than those obtained using the conventional multivariate anomaly detection algorithms. Compared to the classic backpropagation NN-based algorithm, the DBN-based algorithm is notably advantageous in terms of the model training time and convergence.

Fully Written Flexible Potentiometric Sensor Using Two-Dimensional Nanomaterial-Based Conductive Ink.

The emerging demand for flexible, portable, easily accessible, and cost-effective electronic fabrication has motivated the development of novel techniques to manufacture electronic components and devices. Inspired by daily hand-writing, an all-written potentiometric sensor was developed by using a Chinese brush pen-based writing technique. A writing ink made from graphene nanosheet (GN) as a conductive component, Triton X-100 as a stabilizer, and xanthan gum as a binder, was used to obtain flexible electrode substrate. Results demonstrate the GN ink-based writing electrode (GN-WE) possesses good conductivity, fast electron-transfer kinetics, considerable stability, and favorable flexibility. By further writing cadmium ion selective membrane (Cd2+-ISM) and photopolymerized reference membrane (RM) on the surface of GN-WE, an all-solid-state potentiometric sensor for cadmium ion was constructed. A large bulk capacitance (41.67 µF) and excellent potential stability (drift of 0.156 mV h-1) were achieved at the developed all-written potentiometric sensor, which is much superior to the solid-contact potentiometric sensor using GCE as electrode substrate. Furthermore, real sample analysis reveals that our GN ink-based potentiometric sensor could be used as a reliable and stable sensor for cadmium ion detection in food and in the environment.

Metallic Transition Metal Dichalcogenide Nanosheets as an Effective and Biocompatible Transducer for Electrochemical Detection of Pesticide.

Owing to their large specific surface, favorable electrical conductivity, and excellent electrocatalytic capabilities, two-dimensional transition metal dichalcogenides have received considerable attention in the field of biosensors. On the basis of these properties, we developed a portable and disposable enzyme-based biosensor for paraoxon detection using a metallic MoS2 nanosheets modified screen-printed electrode (SPE). The exfoliated ultrathin metallic MoS2 nanosheets can accelerate the electron transfer on electrode surface and contribute to the immobilization of acetylcholinesterase (AChE) via the cross-linking of glutaraldehyde. Electrodeposited gold nanoparticles (AuNPs) on SPE were used to immobilize MoS2 nanosheets through the interaction between Au atoms on AuNPs and S atoms on MoS2. Using acetylcholine as the substrate, AChE can catalyze the formation of electroactive thiocholine and further generate the redox current. In the presence of paraoxon, the activity of AChE can be inhibited, making the related electrochemical signals weaken. Under the optimized conditions, this electrochemical biosensor exhibited a favorable linear relationship with the concentration of paraoxon from 1.0 to 1000 µg L-1, with the detection limit of 0.013 µg L-1. Furthermore, this developed biosensor was successfully applied to detect paraoxon in pretreated apple and pakchoi samples, which can provide a reliable method for the rapid analysis of organophosphorus pesticides in agricultural products.

Associations between pan-immune-inflammation value and abdominal aortic calcification: a cross-sectional study.

Background: Abdominal aortic calcification (AAC) pathogenesis is intricately linked with inflammation. The pan-immune-inflammation value (PIV) emerges as a potential biomarker, offering reflection into systemic inflammatory states and assisting in the prognosis of diverse diseases. This research aimed to explore the association between PIV and AAC. Methods: Employing data from the National Health and Nutrition Examination Survey (NHANES), this cross-sectional analysis harnessed weighted multivariable regression models to ascertain the relationship between PIV and AAC. Trend tests probed the evolving relationship among PIV quartiles and AAC. The study also incorporated subgroup analysis and interaction tests to determine associations within specific subpopulations. Additionally, the least absolute shrinkage and selection operator (LASSO) regression and multivariable logistic regression were used for characteristics selection to construct prediction model. Nomograms were used for visualization. The receiver operator characteristic (ROC) curve, calibration plot and decision curve analysis were applied for evaluate the predictive performance. Results: From the cohort of 3,047 participants, a distinct positive correlation was observed between PIV and AAC. Subsequent to full adjustments, a 100-unit increment in PIV linked to an elevation of 0.055 points in the AAC score (ß=0.055, 95% CI: 0.014-0.095). Categorizing PIV into quartiles revealed an ascending trend: as PIV quartiles increased, AAC scores surged (ß values in Quartile 2, Quartile 3, and Quartile 4: 0.122, 0.437, and 0.658 respectively; P for trend <0.001). Concurrently, a marked rise in SAAC prevalence was noted (OR values for Quartile 2, Quartile 3, and Quartile 4: 1.635, 1.842, and 2.572 respectively; P for trend <0.01). Individuals aged 60 or above and those with a history of diabetes exhibited a heightened association. After characteristic selection, models for predicting AAC and SAAC were constructed respectively. The AUC of AAC model was 0.74 (95%CI=0.71-0.77) and the AUC of SAAC model was 0.84 (95%CI=0.80-0.87). According to the results of calibration plots and DCA, two models showed high accuracy and clinical benefit. Conclusion: The research findings illuminate the potential correlation between elevated PIV and AAC presence. Our models indicate the potential utility of PIV combined with other simple predictors in the assessment and management of individuals with AAC.

A laser-Engraved Wearable Electrochemical Sensing Patch for Heat Stress Precise Individual Management of Horse.

In point-of-care diagnostics, the continuous monitoring of sweat constituents provides a window into individual's physiological state. For species like horses, with abundant sweat glands, sweat composition can serve as an early health indicator. Considering the salience of such metrics in the domain of high-value animal breeding, a sophisticated wearable sensor patch tailored is introduced for the dynamic assessment of equine sweat, offering insights into pH, potassium ion (K+), and temperature profiles during episodes of heat stress and under normal physiological conditions. The device integrates a laser-engraved graphene (LEG) sensing electrode array, a non-invasive iontophoretic module for stimulated sweat secretion, an adaptable signal processing unit, and an embedded wireless communication framework. Profiting from an admirable Truth Table capable of logical evaluation, the integrated system enabled the early and timely assessment for heat stress, with high accuracy, stability, and reproducibility. The sensor patch has been calibrated to align with the unique dermal and physiological contours of equine anatomy, thereby augmenting its applicability in practical settings. This real-time analysis tool for equine perspiration stands to revolutionize personalized health management approaches for high-value animals, marking a significant stride in the integration of smart technologies within the agricultural sector.

HAO2 protects from proximal tubular cells injured in rats with chronic kidney disease by promoting fatty acid metabolic processes.

The complex pathogenesis of kidney disease is closely related to the diversity of kidney intrinsic cells. In this study, single-cell transcriptome sequencing technology was used to sequence and analyze blood and kidney tissue cells in normal control rats and rats with chronic kidney disease (CKD), focusing on key cell populations and functional enrichment to explore the pathogenesis of CKD. Oil red O staining and enzyme-linked immunosorbent assay (ELISA) were used to detect lipid droplets and free fatty acid (FFA). Quantitative real-time polymerase chain reaction (RT-PCR), western blot (WB) were used to verify the differential gene hydroxyacid oxidase 2 (HAO2) and fatty acid metabolic process in tissue to ensure the reliability of single-cell sequencing results. We successfully established a single-cell transcriptome atlas of blood and kidney tissue in rats with CKD, which were annotated into 14 cell subsets (MPCs, PT, Tc, DCT, B-IC, A-IC, CNT, ALOH, BC, Neu, Endo, Pla, NKT, Baso) according to marker gene, and the integrated single-cell atlas of rats showed a significant increase and decrease of MPCs and PTs in the CKD group, respectively. Functional analysis found extensive enrichment of metabolic-related pathways in PT cells, includes fatty acid metabolic process, cellular amino acid metabolic process and generation of precursor metabolites and energy. Immunohistochemical experiments determined that the differential gene HAO2 was localized in the renal tubules, and its expression was significantly reduced in CKD group compared with control, and oil red O staining showed that lipid droplets increased in the CKD group, after overexpression of HAO2 the lipid droplets was inhibited. ELISA assay showed that ATP content decreased in the CKD group and FFA increased in the CKD group. Moreover, the mitochondrial membrane potential of the cells in the OE-HAO2 group was significantly increased compared with OE-NC. The acyl-CoA oxidase 1(ACOX1), peroxisome proliferator-activated receptor alpha (PPARα), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) were decreased in the CKD group, while genes and proteins were increased after overexpression of HAO2, and the AMP-activated protein kinase (AMPK) phosphorylated proteins were increased, the acetyl-CoA carboxylase (ACC) phosphorylated proteins were decreased, reversely. Therefore, HAO2 may be an important regulator of fatty acid metabolic processes in CKD, and overexpression of HAO2 can enhance fatty acid metabolism by promoting fatty acid oxidation (FAO) pathway.

IGJ suppresses breast cancer growth and metastasis by inhibiting EMT via the NF­κB signaling pathway.

Breast cancer metastasis is the primary cause of mortality of patients with breast cancer. The present study aimed to explore the role and underlying mechanisms of IGJ in the invasion and metastasis of breast cancer. The Cancer Genome Atlas database was utilized to analyze the differential gene expression profiles in patients with breast cancer with or without metastasis; the target gene, joining chain of multimeric IgA and IgM (JCHAIN, also known as IGJ, as referred to herein), with significant expression and with prognostic value was screened. The expression levels of IGJ in human breast cancer paired tissues and cell lines were detected using reverse transcription­quantitative PCR and western blot analysis. IGJ differential expression was detected in paired human breast cancer tissues using immunohistochemistry. The role of IGJ in breast cancer was verified using CCK­8, invasion and migration assays, and scratch tests in vivo and in vitro. Further exploration of the role and mechanism of IGJ in breast cancer was conducted through Gene Set Enrichment Analysis, Kyoto Encyclopedia of Genes and Genomes analysis, western blot analysis and immunofluorescence experiments. Through the analysis of gene expression profiles, it was found that IGJ was poorly expressed in patients with breast cancer with metastasis compared to patients with non­metastatic breast cancer. The overexpression of IGJ was associated with an improved distant metastasis­free survival and overall survival (OS). COX multivariate regression analysis demonstrated that IGJ was an independent prognostic factor for the OS and relapse­free survival of patients with breast cancer. In comparison to healthy breast cancer adjacent tissues and cell lines, IGJ was poorly expressed in breast cancer tissues and cell lines (P<0.05). Further analyses indicated that the overexpression of IGJ suppressed the proliferation, invasion and metastasis of breast cancer cells in vivo and in vitro by inhibiting the occurrence of epithelial­to­mesenchymal transition (EMT) and suppressing the nuclear translocation of p65. Finally, rescue experiments indicated that IGJ restricted the proliferation and metastasis of breast cancer cells by regulating the NF­κB signaling pathway. On the whole, the present study demonstrates that IGJ suppresses the invasion and metastasis of breast cancer by inhibiting both the occurrence of EMT and the NF­κB signaling pathway. These findings may provide novel biomarkers and potential therapeutic targets for the treatment of metastatic breast cancer.

Effects of sodium thiosulfate on serum calcification factors in patients undergoing maintenance hemodialysis.

OBJECTIVE: To investigate the effect of sodium thiosulfate (STS) on serum calcification factors in patients undergoing maintenance hemodialysis. METHODS: Forty-four Patients were randomly divided into control group (n = 22) and observation group (n = 22) by envelope method (block 4 randomization). The control group received routine treatment while observation group was treated with STS on the basis of routine treatment. The biochemical indicators, including BUN, UA, SCr, Ca2+ , P3- , calcium-phosphorus product, PTH, hs-CRP, TG, TC, HDL, LDL, and serum calcification factor MGP, FA, FGF-23, and OPG levels were compared before and after treatment. RESULTS: Control group had no statistically significant difference in the levels of vascular calcification factors MGP, FA, FGF-23, and OPG before and after treatment (p > 0.05). Whereas observation group had higher levels of MGP and FA, and lower levels of FGF-23 and OPG after treatment than before treatment (p < 0.05). The levels of MGP and FA in observation group were higher than those in control group, and FGF-23 and OPG were lower than those in control group (p < 0.05). CONCLUSION: It is speculated that sodium thiosulfate can alleviate the progression of vascular calcification by changing the levels of calcification factors.

Implantable Electrochemical Microsensors for In Vivo Monitoring of Animal Physiological Information.

In vivo monitoring of animal physiological information plays a crucial role in promptly alerting humans to potential diseases in animals and aiding in the exploration of mechanisms underlying human diseases. Currently, implantable electrochemical microsensors have emerged as a prominent area of research. These microsensors not only fulfill the technical requirements for monitoring animal physiological information but also offer an ideal platform for integration. They have been extensively studied for their ability to monitor animal physiological information in a minimally invasive manner, characterized by their bloodless, painless features, and exceptional performance. The development of implantable electrochemical microsensors for in vivo monitoring of animal physiological information has witnessed significant scientific and technological advancements through dedicated efforts. This review commenced with a comprehensive discussion of the construction of microsensors, including the materials utilized and the methods employed for fabrication. Following this, we proceeded to explore the various implantation technologies employed for electrochemical microsensors. In addition, a comprehensive overview was provided of the various applications of implantable electrochemical microsensors, specifically in the monitoring of diseases and the investigation of disease mechanisms. Lastly, a concise conclusion was conducted on the recent advancements and significant obstacles pertaining to the practical implementation of implantable electrochemical microsensors.

An Integrated Solar Panel with a Triboelectric Nanogenerator Array for Synergistic Harvesting of Raindrop and Solar Energy.

The triboelectric nanogenerator (TENG) is regarded as an effective strategy for harvesting energy from raindrops, and is a complementary solution with solar cells to achieve all-weather energy harvesting and sustainable energy supply. However, due to the irregularity of natural rainfalls in the volume, frequency, density, and location, designing high-efficiency raindrop TENG (R-TENG) arrays faces great challenges. In this work, a highly transparent, large-area, and high-efficiency R-TENG array with rational material choice, electrode structure, and array distribution is developed for efficiently harvesting irregular raindrop energy. The problem of electrical signal cancellation among adjacent raindrops can be fully avoided, as viewed from the high-resolution space-time analyses of high-speed camera and electrical signal characteristics. With the rationally designed electrode instead of multiple complex electrodes, all charges can be exported by the R-TENG array in a simulated irregular raindrop scenario. Moreover, it is demonstrated that the R-TENG possesses higher average power density (40.80 mW m-2 ) than that of the solar cell (37.03 mW m-2 ) in rainy condition. Additionally, a self-powered wireless light-intensity-monitoring system is demonstrated for real-time and all-day weather monitoring. This work provides useful guidance for designing high-efficiency TENG arrays integrated with solar panels for harvesting irregular raindrop energy and solar energy.

Inflammation promotes erythropoietin induced vascular calcification by activating p38 pathway.

The current research aimed to verify the effects of erythropoietin (EPO) on vascular calcification under inflammatory conditions and the molecular regulator of vascular calcification induced by EPO. To induce vascular calcification and systemic chronic inflammation in SD rats, EPO was administered intraperitoneally, and 10% casein was injected subcutaneously. The administration period lasted for 20 consecutive weeks. Blood samples were subsequently collected to detect inflammatory factors and vascular calcification. Additionally, high-dose EPOs were applied to stimulate primary vascular smooth muscle cells (VSMCs), and vascular calcification was measured using alizarin red staining, alkaline phosphatase (ALP) activity, and calcium salt quantification. The probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) was employed to detect cellular reactive oxygen species (ROS) levels. The expressions of bone formation-related protein and anti-calcification protein matrix gla protein (MGP) were determined via Western blot. Compared with the control group, calcium deposits and vascular calcification were increased in the EPO group, tumor necrosis factor-alpha (TNF-α) group and TNF-α+ EPO group, whereas MGP was significantly reduced. Moreover, under the stimulation of TNF-α and EPO+TNF-α, pp38/p38 was increased substantially, the addition of p38 inhibitor SB203580 could significantly reduce calcium deposits and vascular calcification. In vivo experiment, compared with the EPO group, calcium salt deposition and vascular calcification were elevated in the EPO+casein group. The present results revealed that high-dose EPO could cause calcification of the abdominal aorta in rats. The inflammatory response aggravated the vascular calcification induced by EPO via activating p38 and ROS levels.

Overexpression of TP53INP2 Promotes Apoptosis in Clear Cell Renal Cell Cancer via Caspase-8/TRAF6 Signaling Pathway.

Clear cell renal cell cancer (ccRCC) is a tumor of high malignancy, which can escape apoptosis. The tumor protein p53-inducible nuclear protein 2 (TP53INP2), known as an autophagy protein, is the essential part for autophagosome formation and sensitizes cells to apoptosis. Our study is aimed at exploring the role of TP53INP2 in ccRCC. We have identified the autophagy-related genes (ARGs) of differential expression in ccRCC patients with the help of the TCGA database by bioinformatics analysis. Our assays of quantitative real-time polymerase chain reaction (qRT-PCR) and western blot were for the determination on the both levels of mRNA and protein. Overexpression of TP53INP2 on cellular proliferation, migration, and apoptosis of ccRCC was verified in the ways of performing CCK-8, wound scrape, transwell and flow cytometry assays in vitro, and a mice tumor model in vivo. Transmission electron microscopy was used to measure autophagy formation. The underlying mechanisms of TP53INP2 on ccRCC were determined via coimmunoprecipitation. TP53INP2 was found highly associated with an outcome of worse overall survival (OS) in Kaplan-Meier curves, and this parameter in ccRCC tissues was also lower than the normal tissues. Overexpression of TP53INP2 inhibited ccRCC cellular proliferation, migration, and invasion, as well as the tumor growth of mice. Those cells treated with autophagy inhibitor chloroquine (CQ) or TP53INP2 increased the apoptosis rate. TP53INP2 promoted autophagy formation and elevated the ratio of LC3 II/LC3 I. However, TP53INP2 did not significantly decrease the p-mTOR level. In addition, TP53INP2 activates the expressions of caspase-3, caspase-8, and PARP. Caspase-8 and TNF receptor associated factor 6 (TRAF6) were found to bind to each other in the presence of TP53INP2. TP53INP2 induces apoptosis in ccRCC cells through caspase-8/TRAF6 pathway, rather than the autophagy-dependent pathway.

Identification of Pathologic Grading-Related Genes Associated with Kidney Renal Clear Cell Carcinoma.

Background: Renal epithelium lesions can cause renal cell carcinoma. This kind of tumor is common among all renal cancers with poor prognosis, of which more than 70% belong to kidney renal clear cell carcinoma. As the pathogenesis of KIRC has not been elucidated, it is necessary to be further explored. Methods: The Genomic Spatial Event database was used to obtain the analysis dataset (GSE126964) based on the GEO database, and The Cancer Genome Atlas was applied for KIRC data collection. edgeR and limma analyses were subsequently conducted to identify differentially expressed genes. Based on the systems biology approach of WGCNA, potential biomarkers and therapeutic targets of this disease were screened after the establishment of a gene coexpression network. GO and KEGG enrichment used cluster Profiler, enrichplot, and ggplot2 in the R software package. Protein-protein interaction network diagrams were plotted for hub gene collection via the STRING platform and Cytoscape software. Hub genes associated with overall survival time of KIRC patients were ultimately identified using the Kaplan-Meier plotter. Results: There were 1863 DEGs identified in total and ten coexpressed gene modules discovered using a WGCNA method. GO and KEGG analysis findings revealed that the most enrichment pathways included Notch binding, cell migration, cell cycle, cell senescence, apoptosis, focal adhesions, and autophagosomes. Twenty-seven hub genes were identified, among which FLT1, HNRNPU, ATP6V0D2, ATP6V1A, and ATP6V1H were positively correlated with OS rates of KIRC patients (p < 0.05). Conclusions: In conclusion, bioinformatic techniques can be useful tools for predicting the progression of KIRC. DEGs are present in both KIRC and normal kidney tissues, which can be considered the KIRC biomarkers.