The development of a novel zeolite-based assay for efficient and deep plasma proteomic profiling.

Plasma proteins are considered the most informative source of biomarkers for disease diagnosis and monitoring. Mass spectrometry (MS)-based proteomics has been applied to identify biomarkers in plasma, but the complexity of the plasma proteome and the extremely large dynamic range of protein abundances in plasma make the clinical application of plasma proteomics highly challenging. We designed and synthesized zeolite-based nanoparticles to deplete high-abundance plasma proteins. The resulting novel plasma proteomic assay can measure approximately 3000 plasma proteins in a 45 min chromatographic gradient. Compared to those in neat and depleted plasma, the plasma proteins identified by our assay exhibited distinct biological profiles, as validated in several public datasets. A pilot investigation of the proteomic profile of a hepatocellular carcinoma (HCC) cohort identified 15 promising protein features, highlighting the diagnostic value of the plasma proteome in distinguishing individuals with and without HCC. Furthermore, this assay can be easily integrated with all current downstream protein profiling methods and potentially extended to other biofluids. In conclusion, we established a robust and efficient plasma proteomic assay with unprecedented identification depth, paving the way for the translation of plasma proteomics into clinical applications.

Stability Analysis in Milling Based on the Localized Differential Quadrature Method.

Chatter stability analysis is an effective way to optimize the cutting parameters and achieve chatter-free machining. This paper proposes a milling chatter stability analysis method based on the localized differential quadrature method (LDQM), which has the advantages of simple principle, easy application, and high computational efficiency. The milling process, considering the regeneration effect, is modeled using linear periodic delay differential equations (DDE), then the state transition matrix during the adjacent cutting period is constructed based on the LDQM, and finally, the stability of the milling process is obtained based on the Floquet theory. The accuracy and computation efficiency of the proposed method are verified through two benchmark milling models. The simulation results demonstrate that the proposed method in this paper can accurately and quickly obtain the chatter stability lobe diagram (SLD) of the milling process, which will provide guidance for optimizing the process parameters.

A Study on the Effect of Nursing Intervention Based on Health Behavior Change Integration Theory on Patients with Limb Fracture and Its Effect on Limb Function and Self-Efficacy.

Objective: To assess the impact on nursing care based on the theory of health behavior change integration in patients with limb fracture and its effect on limb function and self-efficacy. Methods: The starting and ending time of this study is from February 2021 to February 2022. In this study, 116 hospitalized patients with limb fracture were selected as the object of study. According to the method of random number table, the patients were divided into study group and control group. The patients of the study cohort were given nursing healthcare based on the theory of health behavior change integration, while those in the control cohort were given common nursing. The scores of self-rating anxiety scale (SAS), self-rating depression scale (SDS), Frankl compliance scale, self-efficacy scale (GSES), activity of daily living scale (ADL), and Fugl-Meyer limb motor function assessment (FMA) were studied before and 3 months after nursing. Results: After 3 months of nursing, the SAS and SDS scores of the study group were lower than those of the control group, and the difference between groups was statistically significant (p < 0.05). After 3 months of nursing, the scores of Frankl scale, GSES, ADL scale, and FMA scale in the study group were higher than those in the control group, and the difference between groups was statistically significant (p < 0.05). Conclusion: The value of nursing interventions based on the integration of health behavior change theory is even more significant in patients with limb fractures. It is more helpful in reducing patients' anxiety and depression, increasing compliance and improving limb function. Self-efficacy and daily living skills were also significantly improved.

A Prognostic Model for Colon Adenocarcinoma Patients Based on Ten Amino Acid Metabolism Related Genes.

Background: Amino acid metabolism plays a vital role in cancer biology. However, the application of amino acid metabolism in the prognosis of colon adenocarcinoma (COAD) has not yet been explored. Here, we construct an amino acid metabolism-related risk model to predict the survival outcome of COAD and improve clinical decision making. Methods: The RNA-sequencing-based transcriptome for 524 patients with COAD from The Cancer Genome Atlas (TCGA) was selected as a training set. The integrated Gene Expression Omnibus (GEO) dataset with 1,430 colon cancer samples was used for validation. Differential expression of amino acid metabolism-related genes (AAMRGs) was identified for prognostic gene selection. Univariate cox regression analysis, LASSO-penalized Cox regression analysis, and multivariate Cox regression analysis were applied to construct a prognostic risk model. Moreover, the correlation between risk score and microsatellite instability, immunotherapy response, and drug sensitivity were analyzed. Results: A prognostic signature was constructed based on 10 AAMRGs, including ASPG, DUOX1, GAMT, GSR, MAT1A, MTAP, PSMD12, RIMKLB, RPL3L, and RPS17. Patients with COAD were divided into high-risk and low-risk group based on the medianrisk score. Univariate and multivariate Cox regression analysis revealed that AAMRG-related signature was an independent risk factor for COAD. Moreover, COAD patients in the low-risk group were more sensitive to immunotherapy targeting PD-1 and CTLA-4. Conclusion: Our study constructed a prognostic signature based on 10 AAMRGs, which could be used to build a novel prognosis model and identify potential drug candidates for the treatment of COAD.

Mechanical Performance of RPC and Steel-RPC Composite Structure with Different Fiber Parameters: Experimental and Theoretical Research.

This paper aims to explore the material properties of RPC and transverse-bending performance, as well as the crack-width-calculation theory of a densely reinforced steel-RPC composite structure with different fiber parameters. Two fiber types (straight fiber, hybrid fiber) and four fiber volume contents (2%, 2.5%, 3%, 3.5%) were selected to explore the mechanical properties of RPC materials, and the influences of fiber parameters on compressive strength, modulus of elasticity, flexural strength and axial tensile property were investigated. Eight steel-RPC composite plates with different design parameters (fiber type and reinforcement ratio) were conducted to study the transverse-bending performance of steel-RPC composite deck structures. The results show that the addition of 3.5% hybrid fibers to the RPC matrix leads to the optimum axial tensile and flexural properties. Furthermore, the failure mode, load-displacement curve, crack occurrence and propagation characteristics of the composite structure are analyzed in detail. Based on the experimental results, the calculation methods of reinforcement stress and crack width of densely reinforced steel-RPC composite structure are proposed. The calculated results of reinforcement stress and maximum crack width are in good agreement with the actual measured values, which can provide a reference for engineering design.

DNA hydrogels as selective biomaterials for specifically capturing DNA, protein and bacteria.

The ability to selectively capture biomacromolecules and other components from solution has many important applications in biotechnology. However, capturing targets from solution while minimizing interference with the sample solution is still challenging. Here, we describe the design and assembly of a group of DNA hydrogels consisting of long single-stranded DNA produced by rolling amplification reaction (RCA) and crosslinked by DNA duplexes. The developed DNA hydrogels can selectively capture and separate oligonucleotides, proteins and bacteria from solution in situ without complex separation processes. Since such DNA hydrogels can capture their targets in the solution independently, multiple DNA hydrogels that target different compounds can be employed to separate different compounds in the solution at the same time. The work not only expands the application of DNA hydrogels, but also paves the way for developing novel selective biomaterials. STATEMENT OF SIGNIFICANCE: Biomaterials capable of selectively capturing various components have great potential in the field of biotechnology. Here, we proposed a new class of hydrogel composed of crosslinked long DNA strands for selectively capturing DNA, protein and bacteria. Unlike traditional polymeric hydrogels that have small meshes and limit macromolecule diffusion owing to the short distance between two adjacent crosslinks, the described DNA hydrogel has a much larger distance between its crosslinks because of the sequence designability of DNA, which allows easy diffusion of biomacromolecules through its networks and greatly expand its specific surface area. Moreover, the developed DNA hydrogel can also easily combine different aptamers to target different components via the Watson-Crick base pairing without making significant changes in its original design.

Suppressing Alpha-Hemolysin as Potential Target to Screen of Flavonoids to Combat Bacterial Coinfection.

The rapid emergence of bacterial coinfection caused by cytosolic bacteria has become a huge threat to public health worldwide. Past efforts have been devoted to discover the broad-spectrum antibiotics, while the emergence of antibiotic resistance encourages the development of antibacterial agents. In essence, bacterial virulence is a factor in antibiotic tolerance. However, the discovery and development of new antibacterial drugs and special antitoxin drugs is much more difficult in the antibiotic resistance era. Herein, we hypothesize that antitoxin hemolytic activity can serve as a screening principle to select antibacterial drugs to combat coinfection from natural products. Being the most abundant natural drug of plant origins, flavonoids were selected to assess the ability of antibacterial coinfections in this paper. Firstly, we note that four flavonoids, namely, baicalin, catechin, kaempferol, and quercetin, have previously exhibited antibacterial abilities. Then, we found that baicalin, kaempferol, and quercetin have better inhibitions of hemolytic activity of Hla than catechin. In addition, kaempferol and quercetin, have therapeutic effectivity for the coinfections of Staphylococcus aureus and Pseudomonas aeruginosa in vitro and in vivo. Finally, our results indicated that kaempferol and quercetin therapied the bacterial coinfection by inhibiting S. aureus α-hemolysin (Hla) and reduced the host inflammatory response. These results suggest that antitoxins may play a promising role as a potential target for screening flavonoids to combat bacterial coinfection.

Analysis of Immune Landscape Reveals Prognostic Significance of Cytotoxic CD4+ T Cells in the Central Region of pMMR CRC.

BACKGROUND: Mismatch repair proficient colorectal cancer (pMMR CRC) lacks effective treatments and has a poor prognosis, which can be attributed to the complexity of tumor microenvironment. The coordinated function of immune cells is vital to anti-tumor immunity. However, the spatial characteristics of immune cells in the pMMR CRC immune microenvironment and their relationship with clinical prognosis are not fully understood. Meanwhile, the immune modulatory effect of neoadjuvant chemotherapy (NCT), which is the first-line treatment of pMMR CRC, needs further investigation. Therefore, this study aims to explore the spatial dynamics of immune cells and its prognostic value in pMMR CRC. METHODS: We analyzed the various immune cells in formalin-fixed, paraffin-embedded tumor tissues which were collected from 77 patients with stage II/III of pMMR CRC, including 39 non-NCT treated and 38 NCT treated patients. We used the optimized multiplex immunohistochemistry (mIHC) to identify and quantify the density, type and location of immune cells in pMMR CRC. Multivariate survival analysis was performed to assess the relationship of immune profiles and clinical prognosis of pMMR CRC patients. RESULTS: The densities of most T cell subsets, B cells and macrophages were higher in the central region of the pMMR CRC than in the invasion margin. Tumor infiltrating lymphocytes (TILs), especially the infiltration of CD4+ GzmB+ T cells in the central region of the tumor was identified to be positively correlated with the prognosis of the patients. Multivariate analysis confirmed that CD4+ GzmB+ T cells population was an independent predictor of disease-free survival (DFS) in non-NCT group. Meanwhile, NCT enhanced the infiltration of CD4+ GzmB+ T cells in the central region of the pMMR CRC, which was also identified as an independent protective factor of overall survival (OS) and DFS in NCT group. CONCLUSION: We demonstrated that the level of CD4+ GzmB+ T cells located in the center of tumor could provide great prognostic value for pMMR CRC patients. And the application of neoadjuvant chemotherapy further improves the infiltration of CD4+ GzmB+ T cells in the central compartment. Further studies into the application of CD4+ GzmB+ T cells in tumor immunotherapy are needed.

Lipid Metabolism in Tumor-Infiltrating T Cells.

T cells recognize "foreign" antigens and induce durable humoral and cellular immune responses, which are indispensable for defending pathogens, as well as maintaining the integrity and homeostasis of tissues and organs. T cells are the major immune cell population in the tumor microenvironment which play a critical role in the antitumor immune response and cancer immune surveillance. Defective immune response of tumor-infiltrating T cells is the main cause of cancer immune evasion. The antitumor response of T cells is affected by multiple factors in the tumor microenvironment, including immunosuppressive cells, immune inhibitory cytokines, tumor-derived suppressive signals like PD-L1, immnuogenicity of tumor cells, as well as metabolic factors like hypoxia and nutrient deprivation. Abundant studies in past decades have proved the metabolic regulations of the immune response of T cells and the tumor-infiltrating T cells. In this chapter, we will discuss the regulations of the antitumor response of tumor-infiltrating T cells by lipid metabolism, which is one of the main components of metabolic regulation.

Potentiating CD8+ T cell antitumor activity by inhibiting PCSK9 to promote LDLR-mediated TCR recycling and signaling.

Metabolic regulation has been proven to play a critical role in T cell antitumor immunity. However, cholesterol metabolism as a key component of this regulation remains largely unexplored. Herein, we found that the low-density lipoprotein receptor (LDLR), which has been previously identified as a transporter for cholesterol, plays a pivotal role in regulating CD8+ T cell antitumor activity. Besides the involvement of cholesterol uptake which is mediated by LDLR in T cell priming and clonal expansion, we also found a non-canonical function of LDLR in CD8+ T cells: LDLR interacts with the T-cell receptor (TCR) complex and regulates TCR recycling and signaling, thus facilitating the effector function of cytotoxic T-lymphocytes (CTLs). Furthermore, we found that the tumor microenvironment (TME) downregulates CD8+ T cell LDLR level and TCR signaling via tumor cell-derived proprotein convertase subtilisin/kexin type 9 (PCSK9) which binds to LDLR and prevents the recycling of LDLR and TCR to the plasma membrane thus inhibits the effector function of CTLs. Moreover, genetic deletion or pharmacological inhibition of PCSK9 in tumor cells can enhance the antitumor activity of CD8+ T cells by alleviating the suppressive effect on CD8+ T cells and consequently inhibit tumor progression. While previously established as a hypercholesterolemia target, this study highlights PCSK9/LDLR as a potential target for cancer immunotherapy as well.

Endothelial-Derived Interleukin-1α Activates Innate Immunity by Promoting the Bactericidal Activity of Transendothelial Neutrophils.

Migration of neutrophils across endothelial barriers to capture and eliminate bacteria is served as the first line of innate immunity. Bacterial virulence factors damage endothelium to produce inflammatory cytokines interacts with neutrophils. However, the mechanisms that behind endothelial-neutrophil interaction impact on the bactericidal activity remain unclear. Therefore, we aimed to find the target proteins on endothelial cells that triggered the bactericidal activity of transendothelial neutrophils. Herein, we built the infected models on rats and endothelial-neutrophil co-cultural system (Transwell) and discovered that endothelial-derived IL-1α promoted the survival of rats under Escherichia coli infection and enhanced the bactericidal activity of transendothelial neutrophils in vivo and in vitro. Results further showed that IL-1α was inhibited by lipopolysaccharide (LPS) in the endothelial-neutrophil interaction. We found that LPS mainly damaged cell membrane and induced cell necrosis to interrupt neutrophil migration from endothelial barrier. Thus, we used the isobaric tags for relative and absolute quantification (iTRAQ) method to identify different proteins of endothelial cells. Results showed that IL-1α targeted cellular plasma membrane, endoplasmic reticulum and mitochondrial envelope and triggered eleven common proteins to persistently regulate. During the early phase, IL-1α triggered the upregulation of cell adhesion molecules (CAMs) to promote neutrophil adhesion, while oxidative phosphorylation was involved in long time regulation to induce transmigration of neutrophils against bacteria. Our results highlight the critical mechanism of endothelial-derived IL-1α on promoting bactericidal activity of transendothelial neutrophils and the findings of IL-1α triggered proteins provide the potentially important targets on the regulation of innate immunity.

Comparison of the Gut Microbiota Between Pulsatilla Decoction and Levofloxacin Hydrochloride Therapy on Escherichia coli Infection.

Gut microbiota serves as a critical indicator for gut health during treatment of pathogenic bacterial infection. Both Pulsatilla Decoction (abbreviated to PD, a traditional Chinese medicine compound) and Levofloxacin Hydrochloride (LVX) were known to have therapeutic effects to intestinal infectious disease. However, the changes of gut microbiota after PD or LVX treatment remain unclear. Herein, this work aimed to investigate the changes of intestinal flora after PD or LVX therapy of Escherichia coli infection in rats. Results revealed that PD exhibited a valid therapeutic approach for E. coli infection via the intestinal protection, as well as the inhibited release of IL-8 and ICAM-1. Besides, PD was beneficial to rebuild the gut microbiota via restoring Bacteroidetes spp in the composition of the gut microbiota. Comparatively, LVX treatment promoted the infection and ravaged gut microbiota by significantly decreasing Bacteroidetes and increasing Firmicutes. These findings not only highlight the mechanism of Chinese herbal formula, but extend the application of PD as veterinary medicine, feed additive and pre-mixing agent for improving the production of animal derived foods.