Chip Analysis Research Articles

P0087 Calprotectin contributes to refractory Crohn’s disease fistula through dysregulation of epithelial wound healing responses

Abstract Background Refractory perianal fistula in Crohn’s disease (CD) still present an unmet clinical need with a high disease burden. Despite increasing interest, pathophysiology leading to the refractory phenotype remains unclear, with only epithelial-to-mesenchymal-transitioning (EMT) commonly mentioned as a contributing factor. Using a systematic approach comparing therapy refractory CD fistula, therapy responsive non-IBD fistula and normal rectum, we aimed to differentiate between general healing responses and dysregulation thereof in refractory CD fistula. Methods Total RNASeq (n=51), single cell RNASeq (n=31) and spatial transcriptomic (n=8) analysis were performed on the internal opening and tract of CD fistula and idiopathic fistula. Results were validated using immunohistochemistry. Functional studies were performed in human adult organoid cultures and intestinal cell lines. Results In the fistula tracts we identified development of a squamous epithelium characterized by expression of WFDC2, keratin 5 and keratin 13. Although some EMT markers were expressed in the squamous epithelium or its intermediates, no activation of a complete EMT program could be detected in any of the tissue types, regardless of the underlying diagnosis. Upstream analysis indicated TNFα, IL6 and TGFβ as inducers of WFDC2+ epithelium, and human adult colon organoids stimulated with these cytokines (but not TGFβ alone) indeed showed a transciptional profile similar to the fistula epithelium, confirming redifferentiation potential. While WFDC2+ epithelium did occur in CD fistula, abundance was strongly decreased compared to non-IBD fistula. In addition, those WFDC2+ cells present had a more pro-inflammatory profile and were morphologically disorganized. Surprisingly, pseudotime analysis indicated epithelial calprotectin (S100A8/9) as a key factor in the aberrant development of WFDC2+ tissue in CD. Normal intestinal epithelium does not express calprotectin, but a mix of cytokines present in fistula induced clear intracellular expression of S100A8/9 in both epithelial cell lines and primary colon organoids. In CD fistula, calprotectin expression was particularly intranuclear, suggesting a transcriptional role. CHIP analysis indeed showed binding of calprotectin to various inflammatory genes and knockdown of calprotectin abrogated expression of these genes supporting a crucial regulatory role. Conclusion These data identify redifferentiation of rectal mucosa to a squamous phenotype as a normal damage response during fistula formation. However, in CD patients, transcriptional effects of calprotectin derail this process, resulting in disorganized inflammatory tissue not amenable for surgical closure and contributing to refractory disease.

Panzerina lanata accelerates methicillin-resistant Staphylococcus aureus eradication by promoting migration and activation of neutrophils.

Panzerina lanata (Lanata) is generally used to treat pustule infection in Inner Mongolia folk medicine and is called "the holy medicine for pustule." However, the pharmacological mechanism of Lanata in treating pustule infection is still unclear. This study aimed to investigate the therapeutic effects of Lanata on skin infection and explore the underlying mechanisms. A skin wound methicillin-resistant Staphylococcus aureus (MRSA) infection mouse model was established to evaluate the healing effect of Lanata on infected wounds. In vitro assays were also conducted to determine the antibacterial activity of Lanata. Flow cytometry and immunohistochemistry were used to dynamically detect the number of neutrophils in the bone marrow, peripheral blood, and MRSA-infected wound. Protein expression in the infected wound skin was detected by a protein chip. Using an air pouch MRSA infection mouse model, the number of neutrophils, reactive oxygen species (ROS) level in neutrophils, and neutrophil extracellular trap (NET) formation were dynamically detected by flow cytometry and immunofluorescence. RNA-seq, RT-qPCR, flow cytometry, ELISA, and CXC chemokine receptor 2 (CXCR2) and P-selectin glycoprotein ligand-1 (PSGL-1) inhibitors were used to explore the mechanism of Lanata in regulating neutrophils. In vitro assays showed that Lanata had no direct antibacterial activity. In skin wound MRSA-infected mouse, Lanata promoted the rapid migration of neutrophils from the bone marrow via peripheral blood to the wound site to eradicate MRSA in the acute stage of infection and accelerate wound healing. Skin protein chip analysis showed that Lanata upregulated CXCR2 and PSGL-1 protein levels in skin wounds. Furthermore, analysis using the air pouch MRSA infection mouse model found that Lanata not only promoted the rapid migration of neutrophils from peripheral blood to the air pouch but also enhanced the activation of neutrophils, including the increase of ROS and the release of NETs, and upregulated the expression of CXCR2, PSGL-1, and myeloperoxidase (MPO) in neutrophils. Inhibition of CXCR2 and MPO significantly attenuated the effect of Lanata on promoting migration and activation of neutrophils. Panzerina lanata resists MRSA infection by promoting migration and activation of neutrophils to rapidly eradicate MRSA.

Fundamental investigations on temperature development in ultra-precision turning

Ultra-precision machining represents a key technology for manufacturing optical components in medical, aerospace and automotive industry. Dedicated single crystal diamond tools enable the production of innovative optical surfaces and components with high dimensional accuracies and low surface roughness values in a wide range of airborne sensing and imaging applications concerning space telescopes, fast steering mirrors, laser communication and high-energy laser systems. Despite the high mechanical hardness of single crystal diamonds, temperature-induced wear of the diamond tools occurs during the process. In order to increase the economic efficiency of ultra-precision turning, the characterisation and interpretation of cutting temperatures are of utmost importance. According to the state-of-the-art, there are no precise methods for online temperature monitoring during the process at the cutting edge with regard to the requirements for resolution accuracy, response time and accessibility to the cutting edge. For this purpose, a dedicated cutting edge temperature measurement system based on ion-implanted boron-doped single crystal diamonds as a highly sensitive temperature sensor for ultra-precision turning was developed. To enable highly sensitive temperature measurements, ion implantation was used for partial and specific boron doping close to the cutting edge of single crystal diamond tools. Within the investigations, a resolution accuracy of 0.29 °C ≤ aR ≤ 0.39 °C could be proven for the developed cutting edge temperature measurement system. In addition, a total measurement uncertainty of uM = 0.098 °C was determined for the sensor accuracy aS in the investigated process area. For a rake angle range of 0° ≤ γ0 ≤ −30°, reaction times of 420 ms ≤ tR ≤ 440 ms were further determined. Using the developed cutting edge temperature measurement system enables a holistic view of the temperature development during ultra-precision machining, whereby a correlation between the measured cutting temperatures and the chip formation mechanisms depending on the applied process parameters could be identified. Within the investigations, the highest measured temperatures of ϑM = 50.18 °C and simulated maximum temperatures of ϑS,max = 183.12 °C were determined at a cutting speed of vc = 350 m/min, a cutting depth of ap = 35 µm as well as a feed of f = 35 µm using a rake angle of γ0 = −30°. The most uniform chips with the smoothest surfaces were identified within the chip analysis using a cutting speed of vc = 50 m/min, a cutting depth of ap = 5 µm and a feed of f = 35 µm with a measured temperature of ϑM = 21.30 °C and a simulated temperature of ϑS = 38.47 °C in the examined finishing area. According to the results, it was also shown that the cutting edge temperature measurement system with ion-implanted diamonds can be used for both electrically conductive and non-conductive materials. This provides the fundamentals for further research works to identify the complex temperature-induced wear behaviour of single crystal diamonds in ultra-precision turning and serves as the basis for self-optimising and self-learning ultra-precision machine tools.

MiR- 146b-5p inhibits Candida albicans-induced inflammatory response through targeting HMGB1 in mouse primary peritoneal macrophages.

Candida albicans (C. albicans) is one of the most common pathogens associated with deep fungal infection, which represents a serious threat to human health. Although high mobility group box 1 (HMGB1) plays a key role in C. albicans infection, its mechanism is unclear. We aimed to explore the regulation of small-molecule non-coding RNA (miRNA) for HMGB1 in C. albicans infection. Mouse primary peritoneal macrophages (MPMs) were isolated successfully. The optimum conditions for C. albicans infection were selected by Western blot and ELISA. The miRNA differential expression profiles of C. albicans infection were screened and verified by 6 miRNA gene chips and qRT-PCR. The direct regulation of the target gene HMGB1 by mmu-miR-146b-5p was confirmed through a dual-luciferase assay. The levels of mmu-miR-146b-5p, HMGB1, inflammatory mediators, p-IKK, IKK, p-IκBα, IκBα and NF-κB p65 were tested by qRT-PCR, Western blot, and ELISA. The nuclear and cytoplasm translocation of HMGB1 and NF-κB p65 were detected by Western blot and laser confocal microscopy. After siHMGB1 transfection, the expression levels of HMGB1, inflammatory mediators, p-IKK, IKK, p-IκBα, IκBα and NF-κB p65 were assessed using Western blot, qRT-PCR and ELISA. In our study, MPMs were successfully extracted and infected with C. albicans at optimum conditions of 1.5×107CFU/mL for 36h. Through miRNA gene chips analysis, 40 differential genes were screened. mmu-miR-146b-5p could directly and negatively regulate the expression and translocation of HMGB1, inhibit the expression of inflammatory mediators, and might participate in the NF-κB signaling pathway in a HMGB1-dependent manner under C. albicans infection. mmu-miR-146b-5p may play an anti-inflammatory role in treating C. albicans infection and provide a novel target for it.

Arntl-induced upregulation of DUSP1 inhibits tumor progression in esophageal squamous cell carcinoma by inactivating ERK signaling

ABSTRACT Background Esophageal squamous cell carcinoma (ESCC) is a primary histological type of esophageal carcinoma with high morbidity. Aryl hydrocarbon receptor nuclear translocator-like (ARNTL) is a circadian clock gene associated with the progression of multiple tumors. However, its roles and mechanisms in ESCC remain unknown. Methods ARNTL expression was analyzed using TCGA database and detected using qRT-PCR, and ARNTL-related pathways were analyzed through GSEA. Cell functional behaviors were assessed in vitro by measuring cell viability, proliferation, and apoptosis. Cell growth in the murine model was investigated through xenograft model and immunofluorescence assays of PCNA and Ki67. The downstream targets of ARNTL were analyzed through sequencing and identified via luciferase report, ChIP, and RNA pull-down analyses. Dual-specificity protein phosphatase-1 (DUSP1) expression was analyzed using GEO datasets and measured using qRT-PCR and western blotting. Protein expression was examined via western blotting. Results ARNTL expression was decreased in esophageal carcinoma and associated with histological types, and elevated expression of ARNTL repressed ESCC cell viability and proliferation and facilitated cell apoptosis. ARNTL upregulation reduced tumor cell growth in murine models and decreased PCNA and Ki67 levels. Furthermore, DUSP1 was downregulated upon ARNTL silencing in ESCC. ARNTL could bind and positively regulate DUSP1 transcription. Additionally, DUSP1 silencing reversed the influences of ARNTL upregulation on cell viability, proliferation, and apoptosis in ESCC cells. ARNTL attenuated the activation of the ERK signaling by decreasing ERK phosphorylation through upregulation of DUSP1. Conclusion ARNTL hinders cell growth and contributes to cell apoptosis by inactivating ERK signaling through transcriptional upregulation of DUSP1 in ESCC.

CALR frameshift mutation detection in myeloproliferative neoplasms by microfluidic chip analysis

Abstract Background CALR mutation analysis is routinely used to diagnose BCR/ABL1-negative myeloproliferative neoplasms. The 2 most common CALR mutations are a 52–base pair (bp) deletion and a 5-bp insertion, which account for approximately 85% of cases. Methods To evaluate our new microfluidic chip assay, we tested CALR mutant and wild-type specimens that were previously analyzed using conventional methods at a reference laboratory. Samples included EDTA-anticoagulated peripheral blood and bone marrow specimens, air dried bone marrow aspirate smears, and formalin-fixed, paraffin-embedded bone marrow sections. CALR exon 9 was PCR amplified using 2 previously published primer pairs and a third unique primer pair designed for our new assay. Amplicons were sized using microfluidic chip analysis. Results Concordance with the reference method was 100% (42/42). Intra-run and inter-run reproducibility were also 100% (3/3 and 3/3, respectively). The limit of detection was confirmed to be 6% mutant alleles. Conclusion We determined that the microfluidic chip assay to detect CALR exon 9 mutations was acceptable for clinical use. Compared with the conventional method, the microfluidic analysis assay benefits from a streamlined workflow, faster turnaround, and a smaller instrument footprint.

Possible involvement of up-regulated salt-dependent glucose transporter-5 (SGLT5) in high-fructose diet-induced hypertension.

Excessive fructose intake causes a variety of adverse conditions (e.g., obesity, hepatic steatosis, insulin resistance and uric acid overproduction). High fructose-induced hypertension is a particularly common and pathologically significant condition induced by excess fructose, but its underlying mechanisms remain unknown. We investigated these mechanisms in 7-week-old male Sprague-Dawley rats fed normal rat food or a diet containing 60% glucose (GLU group) or 60% fructose (FRU group) for 3, 6, or 12 weeks. Daily food consumption was measured to avoid between-group discrepancies in caloric/salt intake, adjusting for feeding amounts. The mean blood pressure of FRU rats was significantly higher (12 weeks GLU: 94.8 ± 3.4 mmHg vs. 12 weeks FRU: 103.7 ± 1.2 mmHg), and fractional sodium excretion was significantly lower (12 weeks GLU: 0.084 ± 0.011% vs. 12 weeks FRU: 0.059 ± 0.08%), indicating that the high-fructose diet caused salt retention. The kidney weight and glomerular surface area were greater in FRU rats (12 weeks GLU: 7495 ± 181 vs. 12 weeks FRU: 9831 ± 164 μm2), suggesting that the high-fructose diet induced an increase in extracellular fluid volume. The expressions of GLUT5 and ketohexokinase, an enzyme required for fructose metabolism, were up-regulated in the FRU group rats (GLUT5 12 weeks GLU: 104.7 ± 15.4% vs. 12 weeks FLU: 309.0 ± 99.9%, ketohexokinase 12 weeks GLU: 129.6 ± 3.5% vs. 12 weeks FLU: 163.9 ± 13.0%). Cortical ATP levels were significantly lower in FRU rats (12 weeks GLU: 9.82 ± 1.26 nmol/mg protein vs. 12 weeks FRU: 7.59 ± 1.68 nmol/mg protein), possibly indicating ATP consumption due to fructose metabolism. Unlike in previous reports the high-fructose diet did not affect NHE3 expression (12 weeks GLU: 166.1 ± 6.3% vs. 12 weeks FLU: 142.0 ± 5.9%). A gene chip analysis conducted to identify susceptible molecules revealed that only Slc5a10 (corresponding to SGLT5) showed >two-fold up-regulation in FRU versus GLU rats. RT-PCR and in situ hybridization confirmed the SGLT5 up-regulation (12 weeks GLU: 75.0 ± 5.8% vs. 12 weeks FLU: 230.1 ± 16.0%). Our findings may indicate that the high-fructose diet increased sodium reabsorption principally through up-regulated SGLT5, finally causing salt-sensitive hypertension.

Fluorescence analysis of wood chips and their constituents

Abstract Micro-spectral analysis and fluorescence decay experiments of beech and oak chips as well as their main constituents lignin and cellulose are reported. While the lignin spectrum shows two fluorescence bands of almost identical intensity around 500 nm and 560 nm, the cellulose spectrum is dominated by a band around 500 with a less pronounced shoulder at 560 nm, whose intensity increases with the amount of residual lignin. Fluorescence decay kinetics are bi-exponential with lifetimes around 0.5 ns and 2.5 ns and an increasing contribution of the short-lived component with an increasing amount of lignin. Both data sets indicate that a quantification of residual lignin in cellulose samples obtained after decomposition of beech wood appears possible in a concentration range of 2–10 %. This may provide a non-destructive and label-free test of the quality of decomposition.

AEBP1 Silencing Protects Against Cerebral Ischemia/Reperfusion Injury by Regulating Neuron Ferroptosis and Microglia M2 Polarization Through PRKCA-PI3K-Akt Axis.

Cerebral ischemia/reperfusion injury is one of the main causes of neuronal damage. Neuron ferroptosis and microglia polarization are considered as critical processes during cerebral ischemia/reperfusion. Adipocyte enhancer-binding protein 1 (AEBP1) usually acts as a transcriptional repressor which is involved in various diseases. However, it is still remains unknown whether AEBP1 could have important roles in regulating the neuron ferroptosis and microglia polarization in cerebral ischemia/reperfusion injury. The oxygen-glucose deprivation and reperfusion (OGD/R)-treated cells and middle cerebral artery occlusion (MCAO)-treated mice were used as in vitro and in vivo models. The differentially expressed factors were analyzed according to GEO datasets. Relative mRNA and protein expression levels were detected by qRT-PCR and western blot analysis. Cell viability was measured by CCK-8 assay. ROS, GSH and iron contents were detected using specifical assay kits. CD26 and CD206 levels were measured by immunofluorescence assay. Inflammatory cytokines were detected by ELISA. The association between AEBP1 and PRKCA was assessed by luciferase reporter and ChIP analyses. The neuron damage in mice was analyzed by TTC staining and neurological deficit score. Transcription factor AEBP1 was increased in OGD/R-treated HT22 and BV2 cells. AEBP1 silencing attenuated OGD/R-induced HT22 cell ferroptosis through increasing cell viability, GSH and GPX4 levels, and decreasing ROS, iron and ACSL4 levels. AEBP1 knockdown promoted microglia M2 polarization by increasing CD206-positive cells and Arg-1 level, and reducing iNOS, TNF-α, IL-1β and IL-6 levels in BV2 cells. AEBP1 transcriptionally repressed PRKCA expression, and further regulated PI3K/Akt signaling activation. Inhibition of PRKCA or PI3K/Akt reversed the effects of AEBP1 silencing on neuron ferroptosis and microglia M2 polarization. AEBP1 downregulation attenuated neuronal damage by decreasing infarct size and deficit scores in MCAO-treated mice. AEBP1 silencing mitigated neuron ferroptosis and promoted microglia M2 polarization through increasing PRKCA and activating PI3K/Akt signaling, indicating the potentially protective action of AEBP1 knockdown in cerebral ischemia/reperfusion injury.

Analysis of single layer artificial neural network neuromorphic hardware chip

The neuromorphic architectures are hardware network systems designed with neural functions. Neural networks seen in biology serve as an inspiration for network systems. A synapse connects every node or neuron in an artificial neural network (ANN) to every other node. As in biological brains, the amplitude of the linking between nodes referred to as synaptic weights will regulate the connection. In contrast to conventional design, ANN uses many highly organized dealing pieces that work together to solve real-world issues. The design of the neuromorphic hardware chip is discussed in the paper. The target device used is a Virtex-5 Field Programmable Gate Array (FPGA) and the simulation is taken on Xilinx ModelSim software. This chip is designed for 20 neuron inputs, each of the neuron inputs is 8-bit. Each 20-neuron input is multiplied by 20 input weights and each weight is 8-bit so when these 20 input weights are multiplied by 20 neuron inputs in the multiplier it gives 16-bit output. A control logic is used in this neuromorphic hardware chip design which is used to feed multiplier output to each input of the hidden layer. The system-level outcome of the hidden layer is then given to the multiplexer which has 20 inputs and one single output. The multiplexer is used to select any of the 20 outputs of the control logic. Finally, to gain an understanding of the performance of this neuromorphic hardware chip, we have computed the hardware utilization parameters. These parameters include slices, input/output blocks (IOBs), registers used, memory, and the overall propagation delay used by the hardware chip.

Study of The Formulation Jicama Flour (Pachyrhizus erosus) and Wheat Flour on The Sensory Characteristics of Brownies Chips

Brownies chips is a meal made from brownies dough that is molded into thin squares and baked dry to produce chips that are easy to consume and last a long time. The aim of this research is to determine the formulation of jicama flour (Pachyrhizus erosus) and wheat flour which produces brownies chips with the best sensory properties on the attributes of color, scent, taste, texture, and overall acceptance. This research was structured in a Complete Randomized Block Design (RAKL) with six treatments and four replications. The formulation treatment of jicama flour and wheat flour consisted of 6 levels, namely T0 (0% : 100%), T1 (10% : 90%), T2 (20% : 80%), T3 (30% : 70%), T4 (40% : 60%), T5 (50% : 50%). Data were analyzed using the Bartlett test and Tukey test, followed by analysis of variance (ANARA), and the Least Significant Difference (BNT) test at the 5% level. Based on the research results, brownies chips with a formulation of jicama flour and wheat flour that produces the best sensory results is the T4 treatment which has a sweet and distinctive chocolate taste, blackish brown color, crunchy and dense texture, and aromatic. typical brownies chips. The results of the chemical analysis of the brownies chips were water content 3.76%, ash content 2.34%, protein content 13.87%, fat content 25.61%, carbohydrate content 54.42%, crude fiber content 6.01%. and calorific value are 503.65 kcal per 100 grams. The bengkuang flour can be used as a substitute up to 40% in the manufacture of brownie chips, producing products with good sensory quality and chemical content and has the potential as an alternative healthy snack.

Temperature Evaluation and Failure Analysis of Wire-Bonded FRD Chips in Surge Conditions

Temperature Evaluation and Failure Analysis of Wire-Bonded FRD Chips in Surge Conditions

M6A RNA Methylation-Mediated TUG1 Stability Maintains Mitochondrial Homeostasis during Kidney Aging by Epigenetically Regulating PGC1-α Expression.

Background: Aging is a significant risk factor for the increased incidence of acute kidney injury and chronic kidney disease, posing significant challenges to global public health. The role of N6-methyladenosine (m6A) in the development of chronic kidney disease has been reported, but the regulatory mechanism of m6A in kidney aging remains unclear. Results: In this study, we identified a long noncoding RNA (lncRNA), called taurine up-regulated 1 (TUG1), which exhibited a significantly decreased level of m6A modification in human aged kidney through the m6A-lncRNA epitranscriptome microarray. Bioinformatics analysis and machine learning predicted that TUG1 had potentially strong interaction with PGC1-α. RNA immunoprecipitation and chromatin immunoprecipitation analysis showed that TUG1 promoted proliferator-activated receptor γ coactivator-1α (PGC1-α) expression by directly interacting with its TUG-1 binding element region, thereby impacting mitochondrial quality control (MQC), cellular senescence, and renal fibrosis. Silencing the RNA m6A methylase methyltransferase 14 (METTL14) or the reader protein insulin-like growth factor 2 mRNA-binding proteins (IGF2BP2) resulted in the weakened stability of lncRNA TUG1, contributing to an imbalance in MQC. Conclusion: Our study demonstrated that the m6A modification and stability of TUG1 were mediated by METTL14 in an IGF2BP2-dependent manner, and modulate the mitochondrial homeostasis in kidney aging by direct targeting PGC-1α. These findings provide a new perspective on potential therapeutic targets for kidney aging. Antioxid. Redox Signal. 41, 993-1013.

Determination of the Minimum Uncut Chip Thickness of Ti-6Al-4V Titanium Alloy Based on Dead Metal Zone.

In Ti-6Al-4V titanium alloy micro-machining, since the uncut chip thickness (UCT) is comparable to the radius of the tool cutting edge, there exists a minimum uncut chip thickness (MUCT), and when the UCT is smaller than the MUCT, the plowing effect dominates the cutting process, which seriously affects the machined surface quality and tool life. Therefore, the reliable prediction of the MUCT is of great significance. This paper used Deform to establish an orthogonal cutting simulation model, studied the effect of the dead metal zone (DMZ) on the micro-cutting material flow, determined the DMZ range, and proposed a new method for determining the MUCT based on the DMZ. Cutting experiments were conducted to verify the accuracy of the simulation model firstly by cutting force, and then confirming the accuracy of the DMZ-based MUCT determination method through chip analysis and surface quality analysis. Finally, the effects of different cutting conditions on DMZ and MUCT were further investigated using the proposed DMZ-based MUCT determination method. The results show that the MUCT of Ti-6Al-4V titanium alloy is 4.833 μm for a tool cutting edge radius of 40 μm and a cutting speed of 10 m/min. The DMZ boundary can be used as the boundary of micro-cutting plastic deformation, and the ratio of MUCT to cutting edge radius, hp/rn will gradually decrease with the increase in the tool cutting edge radius and the cutting speed.

Effect of carbon black properties on cut and chip wear of natural rubber

The effect of carbon black colloidal properties on cut and chip wear of natural rubber compounds is investigated across a wide range of applied impact normal forces using an Instrumented Cut and Chip Analyzer (ICCA). The objective of the study is to determine the basic fatigue and fracture mechanisms that drive cut and chip wear. Natural rubber compounds reinforced with eight different carbon blacks varying in structure and surface area are studied. The loading of the carbon blacks in the rubber compounds is fixed at 50 parts per hundred rubber (phr). The cut and chip performance strongly correlates to both the carbon black morphological properties and the resulting compound mechanical and fracture properties. The cut and chip performance also depends on the applied impact normal force level. At low forces, high structure carbon blacks result in compounds which are stiffer and deflect less under the applied impact normal forces and minimize cut and chip wear. At high forces, low structure carbon black compounds, which are softer and more readily able to crystallize under force-controlled deflection, minimize cut and chip wear. It is argued that at low applied impact normal forces, the cut and chip behavior is dominated by a force-controlled fatigue crack growth mechanism which transitions to a critical tearing energy dominated mechanism at high applied impact normal forces. It is therefore important to understand the severity of application to select optimum compound properties such as the carbon black type to minimize cut and chip wear in application.

Ribosomal protein S3A (RPS3A), as a transcription regulator of colony-stimulating factor 1 (CSF1), promotes glioma progression through regulating the recruitment and autophagy-mediated M2 polarization of tumor-associated macrophages.

Dysregulated expression of ribosomal protein S3A (RPS3A) is associated with the tissue infiltration of immune-related cells in a variety of cancers. However, the role of RPS3A in immune cell infiltration in glioma remains unclear. This study aimed to explore the role of RPS3A in the glioma immune microenvironment.RPS3A expression was detected in tumor tissues from patients with glioma. U251 cells were transfected with RPS3A shRNA (sh-RPS3A) and overexpression vector (pcDNA-RPS3A) and then co-cultured with PMA-induced THP-1 cells. Cell viability, invasion, and apoptosis were detected by Edu staining, Transwell, and flow cytometry, respectively. The expression of tumor-associated macrophage (TAM) M1 and M2 markers was detected with RT-qPCR. Next, the interaction between RPS3A and E4 transcription factor 1 (E4F1) was verified by Co-IP analysis, and the binding of E4F1 to colony-stimulating factor 1 (CSF1) promoter was verified by ChIP analysis. Overexpression vectors of CSF1 and E4F1 were used to treat sh-RPS3A-transfected U251 cells for reversal experiments. Finally, U251 cells transfected with sh-RPS3A adenovirus vectors were subcutaneously injected into nude mice to construct a xenograft tumor model, and the growth and metastasis of glioma in vivo were monitored.RPS3A was significantly upregulated in glioma tissues. Overexpression of RPS3A promoted glioma cell proliferation and invasion and inhibited apoptosis. Moreover, overexpression of RPS3A promoted TAM proliferation, invasion, and M2 polarization. Silencing RPS3A had the opposite effect. Silencing RPS3A inhibited autophagy in U251 cells, whereas rapamycin, an activator of autophagy, reversed the inhibitory effect of RPS3A silencing on TAM M2 polarization. Meanwhile, RPS3A promoted its expression by interacting with E4F1, and E4F1 promoted CSF1 transcriptional activation. Overexpression of CSF1 promoted the proliferation and invasion of U251 cells and reversed the inhibitory effect of RPS3A silencing on TAM proliferation and invasion, but had no effect on TAM M2 polarization. The results of in vivo experiments showed that knockdown of RPS3A significantly inhibited glioma tumor growth and metastasis in vivo.This study revealed that RPS3A recruited TAMs by upregulating E4F1-mediated transcription activation of CSF1, and promoted the M2 polarization of TAMs through autophagy, promoting glioma cell malignant growth and tumor progression.

Target-centric analysis of hepatitis B: identifying key molecules and pathways for treatment

Hepatitis B virus (HBV) poses a significant global health challenge, potentially leading to severe liver conditions, with currently limited effective treatment options available. Xiao-Chai-Hu-Tang (XCHT), a well-known Traditional Chinese Medicine (TCM) prescription, shows promise in clinical trials for treating HBV. Therefore, screening the complex components of XCHT, identifying the active compounds, and closely exploring the targets associated with hepatitis B may constitute an effective strategy for the development of new therapeutic drugs for the treatment of this disease. A systematic pharmacology and GEO chip analysis identified key targets and pathways for hepatitis B treatment and effective ingredients. Molecular docking and molecular dynamics simulation techniques were used to explore the affinity and stability of active compounds with core targets, while assessing the druggability and safety of the active compounds. The therapeutic effect of the active compound protoporphyrin in XCHT on hepatitis B were mediated through key targets such as AKT1, MAPK1, and LCK, as well as key signaling pathways like PI3K-Akt signaling pathway and Ras signaling pathway. Protoporphyrin effectively bond to active pockets of core targets and demonstrated favorable druggability and a high safety threshold. The study provided valuable insights into the development of effective treatments for hepatitis B.

EZH2 Promotes Glioma Cell Proliferation, Invasion, and Migration via Mir-142-3p/KCNQ1OT1/HMGB3 Axis : Running Title: EZH2 Promotes Glioma cell Malignant Behaviors.

This study investigates the role and molecular mechanism of EZH2 in glioma cell proliferation, invasion, and migration. EZH2, miR-142-3p, lncRNA KCNQ1OT1, LIN28B, and HMGB3 expressions in glioma tissues and cells were determined using qRT-PCR or Western blot, followed by CCK-8 assay detection of cell viability, Transwell detection of invasion and migration, ChIP analysis of the enrichment of EZH2 and H3K27me3 on miR-142-3p promoter, dual-luciferase reporter assay and RIP validation of the binding of miR-142-3p-KCNQ1OT1 and KCNQ1OT1-LIN28B, and actinomycin D detection of KCNQ1OT1 and HMGB3 mRNA stability. A nude mouse xenograft model and a lung metastasis model were established. EZH2, KCNQ1OT1, LIN28B, and HMGB3 were highly expressed while miR-142-3p was poorly expressed in gliomas. EZH2 silencing restrained glioma cell proliferation, invasion, and migration. EZH2 repressed miR-142-3p expression by elevating the H3K27me3 level. miR-142-3p targeted KCNQ1OT1 expression, and KCNQ1OT1 bound to LIN28B to stabilize HMGB3 mRNA, thereby promoting its protein expression. EZH2 silencing depressed tumor growth and metastasis in nude mice via the miR-142-3p/KCNQ1OT1/HMGB3 axis. In conclusion, EZH2 curbed miR-142-3p expression, thereby relieving the inhibition of KCNQ1OT1 expression by miR-142-3p, enhancing the binding of KCNQ1OT1 to LIN28B, elevating HMGB3 expression, and ultimately accelerating glioma cell proliferation, invasion, and migration.

Voluntary wheel running exercise rescues behaviorally-evoked acetylcholine efflux in the medial prefrontal cortex and epigenetic changes in ChAT genes following adolescent intermittent ethanol exposure.

Adolescent intermittent ethanol (AIE) exposure, which models heavy binge ethanol intake in adolescence, leads to a variety of deficits that persist into adulthood-including suppression of the cholinergic neuron phenotype within the basal forebrain. This is accompanied by a reduction in acetylcholine (ACh) tone in the medial prefrontal cortex (mPFC). Voluntary wheel running exercise (VEx) has been shown to rescue AIE-induced suppression of the cholinergic phenotype. Therefore, the goal of the current study is to determine if VEx will also rescue ACh efflux in the mPFC during spontaneous alternation, attention set shifting performance, and epigenetic silencing of the cholinergic phenotype following AIE. Male and female rats were subjected to 16 intragastric gavages of 20% ethanol or tap water on a two-day on/two-day off schedule from postnatal day (PD) 25-54, before being assigned to either VEx or stationary control groups. In Experiment 1, rats were tested on a four-arm spontaneous alternation maze with concurrent in vivo microdialysis for ACh in the mPFC. An operant attention set-shifting task was used to measure changes in cognitive and behavioral flexibility. In Experiment 2, a ChIP analysis of choline acetyltransferase (ChAT) genes was performed on basal forebrain tissue. It was found that VEx increased ACh efflux in the mPFC in both AIE and control male and female rats, as well as rescued the AIE-induced epigenetic methylation changes selectively at the Chat promoter CpG island across sexes. Overall, these data support the restorative effects of exercise on damage to the cholinergic projections to the mPFC and demonstrate the plasticity of cholinergic system for recovery after alcohol induced brain damage.

Enhancing machinability in free-cutting duplex brass alloys: Isotropic blocky α phase formation via optimized hot extrusion processing

Despite the active utilization of duplex brass alloys in the electronics and automotive industries, optimizing thermomechanical treatment conditions for achieving excellent machinability remains a subject of ongoing debate. In this study, to establish a strategy to achieve enhanced machinability via industrial-scale direct hot extrusion processing, we systematically investigated the effect of hot extrusion processing conditions on microstructural evolution, mechanical properties, and machinability of duplex brass alloys. In particular, the yield strength was effectively interpreted through the extended Hall-Petch relationship, and the machinability was further elucidated through the evaluation of rotational torque during the drilling test as well as the precise analysis of segmented chips produced in the drilling test. As a result, our microstructural analysis of the as-cast Cu58Zn39Pb3 alloy revealed that the cooling process during industrial-scale casting inevitably results in the formation of the Widmanstätten structured α phase and the heterogeneous dispersion of Pb particles. In contrast, 670 ℃ extruded lower part and 730 ℃ extrudates, which were extruded in the single β region that caused complete dynamic recrystallization behavior, exhibited a uniform and isotropic blocky α structure and uniformly redistributed Pb particles, ultimately leading to superior mechanical properties and machinability. Notably, our findings suggest that achieving an isotropic blocky α structure can be attained at temperatures up to 100 ℃ below the β-transus temperature by optimizing extrusion pressure conditions. This approach offers an efficient methodology for minimizing defects caused by high-temperature extrusion, such as surface oxidation and hot shortness cracking, while simultaneously minimizing processing costs.