Tissue Protein Research Articles

Assessing Chronic Heavy Metal Exposure by Analysis of Human Cataract Specimens and the Relationship to Metabolic Profiles

Purpose With age, the mammalian lens forms successive layers of crystallin protein fibers which infoliate with lens growth and development. As heavy metals generally bind to tissue protein, heavy metals are posited to sequester within the lens with age. Therefore, this study aims to compare heavy metals in human crystalline lens of older adults to known physiologic blood and urine levels and assess the association between concentrations in the lens and metabolic biomarkers. Methods Consecutive lens specimens obtained during cataract surgery by phacoemulsification were subjected to atomic spectrometry for heavy metal content. A one-sample t-test compared heavy metals in lens to known physiologic blood and urine concentrations. Linear regression models assessed the association between heavy metals and biomarkers of metabolic function. Linear discriminant analysis assessed the classification of gender and smoking status based on multiple and individual heavy metals. Results All heavy metal levels were elevated in lens specimens compared to blood and urine with the exception of iron (p < 0.0001). Lens titanium and copper were positively associated with blood-urea nitrogen (Titanium: β ̂ = 1.14, p = 0.04, Copper: β ̂ = 1.12, p = 0.03. Lens copper was positively associated with creatinine ( β ̂ = 1.10; p = 0.02), but negatively associated with glomerular filtration rate ( β ̂ = 0.89; p = 0.02). Lens chromium and lead were positively associated with albumin (Chromium: β ̂ = 1.03, p = 0.03; Lead: β ̂ = 1.02, p = 0.04). Lens nickel was positively associated with bilirubin ( β ̂ = 1.14; p = 0.03). Classification based on multiple or individual heavy metals for gender and smoking status was not statistically significant. Conclusions Our results suggest the human crystalline lens accumulates heavy metals with age and demonstrate the correlation between abnormality of metabolic function and heavy metal deposition in older adult lens.

Metabolism and spatial transcription resolved heterogeneity of glutamine metabolism in cervical carcinoma

BackgroundReprogramming of cellular metabolism is a pivotal mechanism employed by tumor cells to facilitate cell growth, proliferation, and differentiation, thereby propelling the progression of cancer. A comprehensive analysis of the transcriptional and metabolic landscape of cervical squamous cell carcinoma (CSCC) at high resolution could greatly enhance the precision of management and therapeutic strategies for this malignancy.MethodsThe Air-flow-assisted Desorption Electrospray Ionization Mass Spectro-metric Imaging (AFADESI-MSI) and Spatial Transcriptomics techniques (ST) were employed to investigate the metabolic and transcription profiles of CSCC and normal tissues. For clinical validation, the expression of ASCT2(Ala, Ser, Cys transporter 2) was assessed using immune histochemistry in 122 cases of cervical cancer and 30 cases of cervicitis.ResultsThe AFADESI-MSI findings have revealed metabolic differences among different CSCC patients. Among them, the metabolic pathways of glutamine show more significant differences. After in situ detection of metabolites, the intensity of glutamate is observed to be significantly higher in cancerous tissue compared to normal tissue, but the intensity is not uniform. To elucidate the potential factors underlying alterations in glutamine metabolism across tissues, we employ ST to quantify mRNA levels. This analysis unveils significant perturbations in glutamine metabolism accompanied by extensive heterogeneity within cervical cancer tissues. After conducting a comprehensive analysis, it has been revealed that the differential expression of ASCT2(encoded by SLC1A5) in distinct regions of cervical cancer tissues plays a pivotal role in inducing heterogeneity in glutamine metabolism. Furthermore, the higher the expression level of ASCT2, the higher the intensity of glutamate is in the region. Further verification, it is found that the expression of ASCT2 protein in CSCC tissues is significantly higher than that in normal tissues (105/122, 86.07%).ConclusionsThis finding suggests that the variation in glutamine metabolism is not uniform throughout the tumor. The differential expression of ASCT2 in different regions of cervical cancer tissues seems to play a key role in causing this heterogeneity. This research has opened up new avenues for exploring the glutamine metabolic characteristics of CSCC which is essential for developing more effective targeted therapies.

Single cell transcriptomics profiling of the stromal cells in the pathologic association of ribosomal proteins in the ischemic myocardium and epicardial fat.

Sustenance of ischemia in the surviving cardiac tissue following myocardial infarction (MI) elicits a proinflammatory milieu resulting in subsequent pathological episodes. Also, the activation and release of ribosomal proteins under ischemic insults have been unveiled; however, their extra ribosomal functions are unknown. We identified the ribosomal proteins including RPL10A, RPL14, RPL30, RPS18, FAU-40 (RPS30), and RPSA (Laminin Receptor, LR) in the vesicles of ischemia challenged epicardial adipose tissue derived stromal cells (EATDS). The present study aimed to assess the association of these proteins in the epicardial adipose tissues (EAT) and left ventricular (LV) myocardium and isolated stromal cells (EATDS and LVSCs) from hyperlipidemic (HL), MI and coronary artery bypass graft (CABG) swine models. The findings revealed an upregulation of RPL10A, RPL14, RPL30, RPS18, RPS30, and RPSA in the LV tissues of CABG and HL swine with a concomitant reduction in the MI group. RPS30 displayed similar upregulation in EAT, whereas the expression of other ribosomal proteins was not significantly altered. Additionally, the ischemic LVSCs and EATDS displayed altered expression status of these genes compared to the control. Also, the RPS18 + , RPL30 + and RPSA + LVSCs favored ischemia and revealed similar anti-inflammatory and regenerative sub-phenotypes reflecting the protective/survival mechanisms. Further understanding regarding the underlying molecular mechanisms and functions of these ribosomal proteins offers immense translational opportunities in the better management of ischemic cardiac complications.

Expression and clinical significance of Numb and Notch-1 proteins between tissue of colon cancer and regional lymph node metastases

ObjectiveTo investigate the expression and clinical significance of Notch-1 and Numb protein in colon cancer tissues and regional lymph node metastases.MethodsImmunohistochemical method was used to detect the expression of Notch-1 protein and Numb protein in 110 cases of colon cancer tissues, along with tumor adjacent tissues and 56 cases of MLN tissues, and to analyze its role in colon cancer and MLN tissue.ResultsComparing colon cancer tissue or lymph node metastases with tumor adjacent tissue, the positive expression rate of Numb was significantly decreased, while the positive expression of Notch-1 was significantly increased in colon cancer tissue or lymph node metastases (both p&amp;lt;0.05). The expression of Notch-1 and Numb was correlated with the lymph node metastasis, TNM stage, and degree of differentiation (p&amp;lt;0.05). The expression between Numb and Notch-1 showed negative correlation in colon cancer tissues (r=−0.261, p&amp;lt;0.05). There was no relationship between the expression of Numb and Notch-1 protein in colon cancer and metastatic lymph node tissue (p&amp;gt;0.05).ConclusionNumb expression is decreased and Notch-1 expression is increased in colon cancer tissue and metastatic lymph node tissue, suggesting that the interaction between the two proteins may play a promote role in the development, invasion, and metastasis of colon cancer. There was no relationship between the expression of Numb and Notch-1 protein in colon cancer and metastatic lymph node tissue, suggesting that there is no obvious enhancement of the cancer cells; in the process of lymph node metastasis, the degree of malignant biological behavior remains relatively stable.

Ultrasound-targeted sirolimus-loaded microbubbles improves acute rejection of heart transplantation in rats by inhibiting TGF-β1-Smad signaling pathway, promoting autophagy and reducing inflammation

Acute rejection (AR) remains a pivotal complication and leading cause of mortality within the first year following heart transplantation (HT). In this study, we assessed the impact of ultrasound-targeted microbubbles loaded with sirolimus (SIR-MBs) on AR in a rat HT model and delved into the underlying mechanisms. We established a rat abdominal ectopic HT model, which was stratified into three groups receiveing the PBS, SIR-MBs + ultrasound-targeted microbubble destruction (UTMD), and sirolimus, respectively. The protective effects of each treatments on survival rate, inflammatory response, autophagy and TGF-β1-Smad signaling pathway-related proteins were evaluted. Additionally, rescue experiment was performed via adding the autophagy inhibitor or TGF-β1 agonist in combination therapy. UTMD combined SIR-MBs mediated 15-fold higher local drug concentration compared to direct sirolimus administration. The infiltration of inflammatory cells in the transplanted hearts indicated that SIR-MBs combined with UTMD were effective in mitigating the inflammatory response, achieving levels significantly lower than those observed in the sirolimus group. Furthermore, after SIR-MBs combined with UTMD treatment, the expression levels of TGF-β1-Smad signaling pathway-related proteins in heart tissues also showed a significant decrease compared to the model control group. Conversely, the expressions of autophagy proteins LC3-II, Beclin-1 and β-arrestin showed an up-regulated trend. Rescue experiments also revealed that the enhancement in survival trends was markedly suppressed following the administration of CsA or SRI-011381, respectively. Collectively, our findings suggest that SIR-MBs combined with UTMD augment the local treatment efficacy for AR in rat HT models by inhibiting the TGF-β1-Smad signaling pathway, promoting autophagy, and alleviating inflammation.

MPO interacts with hRSV particles, contributing to the virucidal effects of NETs against clinical and laboratory hRSV isolates

Human Respiratory syncytial virus (hRSV) mainly affects immunosuppressed patients requiring hospitalization. No specific treatment is financially accessible, and available vaccines do not cover all risk groups. During hRSV infection, there is a robust neutrophilic influx into the airways. hRSV-activated neutrophils release substantial neutrophil extracellular traps (NETs) in lung tissue, comprising DNA, histones, cytosolic, and granular proteins. NETs form mucus buildup in the lungs, compromising respiratory capacity and neutralizing viral particles. Understanding responsible NETs molecules requires improvement. We evaluated NETs interacting with hRSV particles and their contribution to anti-hRSV NET effects. Immunoblotting, immunoprecipitation, and peptide sequencing assays confirmed hRSV binding to a 50–75 kDa NET protein, Myeloperoxidase (MPO). MPO, a microbicide enzyme in NETs, interacts with hRSV, likely at F0 protein (site IV) on the viral surface. Additionally, MPO (32 μM) and NETs (0.4 μg/mL) reduced in vitro replication of clinical (hRSV A and B) and laboratory (Long) hRSV isolates by approximately 30 %, reversible by selective MPO inhibitor (PF-06281355; 48 μM). Thus, MPO contributes to virucidal NET effects on diverse hRSV strains, enhancing comprehension of NETs' role in infection and aiding treatment strategies for respiratory diseases.

Therapeutic efficacy of CRISPR RNA nanoparticles on cervical cancer disease using ultrasound elastography imaging

This work aimed to evaluate the therapeutic efficacy of clustered regularly interspaced short palindromic repeats (CRISPR) ribonucleic acid (RNA) nanoparticles (NPs) in the therapy of cervical cancer (CC) using ultrasound elastography imaging. Using poly(β-amino esters) (PBAE), CRISPR RNA NPs encapsulating Human Papillomavirus 16 (HPV16) E7, namely Poly(β-amino ester) (PBAE)/CRISPR16E7-GFP and PBAE/shRNA-16E7, were prepared. The characterization of these NPs was conducted using transmission electron microscopy (TEM) and laser diffraction particle size analysis. The cytotoxicity of PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 NPs on CC cell lines SiHa, HeLa, and CaSki was assessed using the cell counting kit-8 (CCK-8) assay. Sixty female Sprague Dawley (SD) rats were rolled into six groups randomlyas follows: the blank Ctrl group (Group A, no treatment), the model group (Group B, CC model), the pSpCas9-GFP plasmid group (Group C, CC model + pSpCas9-GFP), the PBAE/CRISPR-16E7 group (Group D, CC model + PBAE/CRISPR-16E7), the pSIREN-U6-shRNA-CMV-iRFP plasmid group (Group E, CC model + pSIREN-U6-shRNA-CMV-iRFP), and the PBAE/shRNA-16E7 group (Group F, CC model + PBAE/shRNA-16E7), each consisting of 10 rats. The impact of NPs on the expression levels (ELs) of HPV16E7 and RB1 proteins in tumor tissues was assessed using Western Blot (WB) analysis. Additionally, ultrasound elastography imaging was employed to analyze the strain ratio (SR) and shear wave velocity (SWV) in lesions of various rat groups. The results indicated that the PBAE/CRISPR16E7-GFP NPs exhibited a particle size of 125.64 ± 7.8 nm, a zeta potential of 19.17 ± 3.61 mV, and a polydispersity index (PDI) of 0.281 ± 0.03. These NPs demonstrated a regular spherical structure with a compact morphology. The PBAE/shRNA-16E7 NPs displayed a particle size of 112.93 ± 9.1 nm, a zeta potential of 13.54 ± 1.39 mV, and a PDI of 0.185 ± 0.03. They exhibited a uniform distribution of spherical shape with regularity in size. The plasmids within PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 NPs exhibited distinct trends of burst and sustained release. The viability of SiHa, HeLa, and CaSki cells remained unaffected by PBAE/CRISPR16E7-GFP NPs. At various mass ratios, neglectable differences (P > 0.05) were observed in cell viability of SiHa, HeLa, and CaSki cells treated with PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 NPs relative to Ctrl group. Relative to Group B, both Groups C and E exhibited a drastic decrease in tumor volume, tumor mass, and HPV16E7 protein ELs (P < 0.05), whereas Groups D and F showed a notably greater reduction in tumor volume, tumor mass, and HPV16E7 protein ELs (P < 0.01). The HPV16E7 protein ELs, SR, and SWV values greatly increased (P < 0.001) in tumors of Group B rats while the EL of RB1 protein notably decreased (P < 0.001) versus Group A. Groups C and E demonstrated elevated HPV16E7 ELs, SR, and SWV values (P < 0.05), with a marked decrease in RB1 protein EL (P < 0.001). Group F exhibited a notable reduction in HPV16E7 protein levels, SR, and SWV values (P < 0.001).The CRISPRRNA NPs composed of PBAE encapsulating HPV16 E7, namely PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 plasmids, demonstrated a significant role in CC therapy. These NPs hold potential application value in the treatment of HPV-related CC, indicating their importance in addressing this particular disease.

Lily extract ameliorate mouse pneumonia by modulating ciliary function

This study explores the biological effects and mechanisms of Lily extract on LPS-induced pneumonia mice. Research has found that after 48 h of treatment with low-dose and high-dose lily extract (100 mg/kg, 200 mg/kg), there was a significant reduction in inflammatory cell infiltration, mRNA expression levels of pro-inflammatory factors (IL-6, IL-1β, TNF-α), protein levels of inflammation-related proteins (IL-6, NF-κB P65) in the lung tissue. In addition, lily extract significantly improved the function of cilia assembly and movement. The expression levels of cilia-related genes (Ttc21a, Cfap45, etc.) and proteins were significantly upregulated. Among them, the high-dose treatment group had tightly arranged cilia and good growth conditions, which was significantly better than the LPS pneumonia model and low-dose treatment group. This found providing novel insights into the pathogenesis and progression of pneumonia. This has important implications for the prevention and treatment of pneumonia, as well as the development of natural agents.

Engrailed-2 (EN2) protein in cervical mucus: a novel biomarker for endometrial carcinoma.

This study aims to demonstrate that the EN2 protein in cervical mucus may serve as a novel biomarker for screening endometrial cancer. This study included 133 patients who were treated at Beijing Obstetrics and Gynecology Hospital. According to the pathological results of hysteroscopy endometrial biopsy, the patients were divided into endometrial cancer group (n = 55), endometrial atypical hyperplasia group (n = 16), benign lesion group (n = 28), and control group (n = 34). All patients collected cervical mucus before hysteroscopy, and the level of EN2 protein in cervical mucus was detected by ELISA in the four groups of patients. Nine patients who underwent surgical treatment for endometrial cancer were included, and endometrial cancer lesions and adjacent tissues without endometrial cancer infiltration were taken from the endometrial cancer lesions and cervical tissues. Fifteen patients who underwent hysterectomy for benign lesions were included, and endometrial and cervical tissues were taken from the endometrial cancer lesions and adjacent tissues without endometrial cancer infiltration. PT-PCR, immunohistochemistry, and WB were used to verify the expression differences of EN2 protein in cancerous lesions and endometrial tissues without endometrial infiltration, cervical tissues, and endometrial tissues without endometrial lesions. In addition, HEC1B, KLE, and HeLa cell lines were used to characterize EN2 overexpression in endometrial cancer cells. Immunohistochemistry, RT-PCR, and confocal analysis were used to detect the expression of EN2 protein in different cell lines. In different groups, the average concentration of EN2 protein in cervical mucus in the EC group was significantly higher than that in the benign group and the normal control group (573.9 ± 123.4ng/mL vs 153.5 ± 106.2ng/mL and 153.0 ± 107.5ng/mL, P < 0.001). The concentration of EN2 protein in EIN3 case specimens was significantly higher than that in the normal control group (P < 0.001). ROC analysis showed that the optimal threshold for diagnosing endometrial cancer in cervical mucus was 321.1ng/ml, with a sensitivity of 92.6% and specificity of 95.4% for distinguishing EINIII, EC, and non-cancerous cases. In clinical specimens, the expression level of EN2 protein in endometrial cancer tissues was significantly higher than that in endometrial tissues and cervical tissues without endometrial cancer infiltration. In addition, PT-PCR, immunohistochemistry, suggest that EN2 protein is overexpressed in endometrial cancer cell lines compared to cervical adenocarcinoma cells (P < 0.01). The concentration of EN2 protein in cervical mucus can effectively identify endometrial cancer and precancerous lesions. The increased expression of EN2 protein in endometrial cancer lesions leads to an increase in the concentration of EN2 protein in the cervical mucus of endometrial cancer patients.This is a small single-center study, and larger studies are needed to determine the role of EN2 protein in the diagnosis of endometrial cancer.

HeMiTo-dynamics: a characterisation of mammalian prion toxicity using non-dimensionalisation, linear stability and perturbation analyses.

Prion-like proteins play crucial parts in biological processes in organisms ranging from yeast to humans. For instance, many neurodegenerative diseases are believed to be caused by the production of prion-like proteins in neural tissue. As such, understanding the dynamics of prion-like protein production is a vital step toward treating neurodegenerative disease. Mathematical models of prion-like protein dynamics show great promise as a tool for predicting disease trajectories and devising better treatment strategies for prion-related diseases. Herein, we investigate a generic model for prion-like dynamics consisting of a class of non-linear ordinary differential equations (ODEs), establishing constraints through a linear stability analysis that enforce the expected properties of mammalian prion-like toxicity. Furthermore, we identify that prion toxicity evolves through three distinct phases for which we provide analytical descriptions using perturbation analyses. Specifically, prion-toxicity is initially characterised by the healthy phase, where the dynamics are dominated by the healthy form of prions, thereafter the system enters the mixed phase, where both healthy and toxic prions interact, and lastly, the system enters the toxic phase, where toxic prions dominate, and we refer to these phases as HeMiTo-dynamics. These findings hold the potential to aid researchers in developing precise mathematical models for prion-like dynamics, enabling them to better understand underlying mechanisms and devise effective treatments for prion-related diseases.

IL-17A-Induced Redox Imbalance and Inflammatory Responses in MiceLung via Act1-TRAF6-IKBα Signaling Pathway: Implications for Lung Disease Pathogenesis.

IL-17A is a potent proinflammatory cytokine that plays a crucial role in the pathogenesis of various lung diseases. This study focused on the evaluation of the role of IL-17 receptor signaling through one-week intranasal exposure of IL-17A in lung tissues of BALB/c mice. IL-17A triggered inflammatory responses in the mice lungs and led to changes in the morphological alveolar arrangements. Exposure of IL-17A induced redox imbalance by triggering an increase in the level of the pro-oxidants (reactive oxygen species, nitrite and malondialdehyde) and reduction of the levels of antioxidant proteins (glutathione, superoxide dismutase and catalase) in the lung tissue. IL-17A also caused a significant elevation in the levels of proinflammatory cytokines lines including TNF-α, IL-1β and IL-6, in lung tissue as well as in plasma. More interestingly, these changes were accompanied by the alterations in IL-17 receptor downstream signaling through activation of IL-17R-Act1-TRAF6-IKBα-mediated pathway. IL-17A exposure also caused lung tissue injury, recruitment and polarization of immune cells from anti-inflammatory to pro-inflammatory. This study clearly demonstrated the role of IL-17A-induced signaling in worsening lung inflammatory diseases, and hence points towards its emergence as an important therapeutic target to control lung inflammation.

Comprehensive Analysis Reveals That ISCA1 Is Correlated with Ferroptosis-Related Genes Across Cancers and Is a Biomarker in Thyroid Carcinoma

Background: ISCA1 (Iron–Sulfur Cluster Assembly 1) is involved in the assembly of iron–sulfur (Fe–S) clusters, which are vital for electron transport and enzyme activity. Some studies suggest the potential involvement of ISCA1 in tumor progression through interactions with ferroptosis-related genes (FRGs) and the tumor immune microenvironment (TME). However, there has been no systematic analysis of its role in FRGs and the TME or its predictive value for prognosis and immunotherapy response across different cancer types. Methods: In this study, we analyzed the expression and prognosis of ISCA1 RNA, CNV, methylation, and protein in multiple tumor tissues via data from the TCGA and CPTAC databases and clinical information. We conducted a comprehensive analysis of the correlations between ISCA1 and FRGs, immune-related genes (including immune regulatory genes and immune checkpoint genes), immune cell infiltration, immune infiltration scores, tumor stemness, and genomic heterogeneity. Results: We performed drug prediction and validation through molecular docking and molecular dynamics analysis to identify candidate drugs that could promote or inhibit ISCA1 RNA expression. Our findings revealed that ISCA1 could serve as a biomarker in thyroid carcinoma, play a role with different FRGs in various cell types, and mediate different ligand–receptor pathways for cell–cell communication. Conclusions: Overall, our study highlights the potential of ISCA1 as a novel biomarker for predicting prognosis and immunotherapeutic efficacy in thyroid carcinoma and suggests its potential for developing novel antitumor drugs or improving immunotherapy.

Proteostasis collapse in rat’s lung tissue as one of the effects of European adder (Vipera berus subsp. berus and Vipera berus subsp. nikolskii) envenomation

Abstract Snakebites and their consequences are a growing concern around the globe. The mixture of enzymes, present in snake venom is a great challenge for a snakebite victim’s body. European adder (Vipera berus) is considered as not very harmful snake due to the mild clinical symptoms of envenomation and rare cases of cardiovascular or pulmonary failure. Nevertheless, at the molecular level, many details of V. berus bite (including proteostasis instability) remain unclear. This work was aimed to determine the effect of Eastern Europe typical adder species Vipera berus berus and Vipera berus nikolskii envenomation on the protein homeostasis of rat’s lung tissue. The decrease in total protein concentration, significant redistribution of main protein fractions (including proteolytically active), rise in middle-mass molecules and low molecular weight peptide concentration have been observed during V. b. berus and V. b. nikolskii envenomation, strongly indicating the proteostasis collapse and endogenous intoxication in lung tissue as an effect of European adder snakebite. Moreover, according to the results of experiments, the effect of V. b. nikolskii venom is more harmful to lung tissue protein homeostasis, comparing to V. b. berus, suggesting that the former may represent a greater danger to people. The results of this study may find applications in antivenom development and could be taken into consideration for proper snakebite treatment selection in order to avoid proteome complications in patients.

Zinc oxide nanoparticles mitigate the malignant progression of ovarian cancer by mediating autophagy-dependent ferroptosis

BackgroundPrevious studies have shown that ZnO-NPs induce autophagy and inhibit the malignant progression of ovarian cancer (OC) cells. This study aims to further explore the mechanism of action of ZnO-NPs on OC.MethodsSKOV3 cells were treat with different concentrations of ZnO-NPs and cell proliferation was assessed through EDU staining. A Xenograft tumor model was established and mice were treated with varying doses of ZnO-NPs for 21 days. Tumor volume and the weight of each group of mice were measured, and the expression of KI67 in tumor tissues was analyzed to evaluate tumor proliferation in vivo. The expression of autophagy and ferroptosis-related proteins in cells and tumor tissues was examined through immunofluorescence, ELISA, and western blotting assays. The relationship between ZnO-NPs induced autophagy and ferroptosis was further investigated using the ferroptosis inhibitors Fer-1, autophagy inhibitor 3-MA and siRNA for ATG5 (si-ATG5).ResultsZnO-NPs dose-dependently reduced the proliferation of SKOV3 cells in vitro. In vivo, both high and low doses of ZnO-NPs effectively inhibited the growth of tumor, reduced pathological damage and the expression of KI67 in tumor tissues. Additionally, ZnO-NPs increased the levels of iron, MDA, 4-HNE, oxidized lipid ROS, ATG5, TFR1, ACSL4, LC3, and Beclin1 in cells and tumor tissues, decreased the expression of SOD, GSH-Px, non-oxidized lipid ROS, GPX4, and p62. Transfection with si-ATG5 or treatment with 3-MA significantly weakened these effects of ZnO-NPs in vitro, with si-ATG5 having a stronger weakening effect on the action of ZnO-NPs than 3-MA. However, ferroptosis inhibitor has a lesser impact on the autophagy of ZnO-NPs-treated SKOV3 cells than the effect of autophagy inhibitors and si-ATG5 on the ferroptosis of ZnO-NPs-treated SKOV3 cells.ConclusionZnO-NPs inhibited the malignant progression of SKOV3 cells by inducing autophagy-dependent ferroptosis.

Chronic Hypobaric Hypoxia Stimulates Differential Expression of Cognitive Proteins in Hippocampal Tissue.

Objective: We aimed to determine changes in cognitive function resulting from chronic hypobaric hypoxia through proteomic analysis of hippocampal tissue. We screened cognition-related proteins to provide ideas and directions that could help prevent and treat hypoxia-associated cognitive impairment. Methods: We analyzed hippocampal tissues from mice exposed to high altitudes and control mice using 4 D label-free quantitative proteomics. The data were analyzed by protein quantitative analysis, functional annotation, differential protein screening, clustering analyses, and functional classification and enrichment. Differential protein expression was investigated using targeted quantitative omics based on parallel response monitoring. Results: We identified and quantified 20 target proteins in 12 samples, of which 18 were significant validated proteins that were or might be related to cognitive functions. Signaling pathways that were significantly enriched in differentially expressed proteins were pyrimidine metabolism, 5'-Adenosine Triphosphate-activated protein kinase signaling, phospholipase D signaling, purine metabolism, inflammatory mediator regulation of transient receptor potential channels, hedgehog signaling pathways, dilated cardiomyopathy, platelet activation, insulin resistance, mRNA surveillance pathways, drug metabolism-other enzymes, and drug metabolism-cytochrome P450. Conclusion: Chronic hypoxia alters protein expression in murine hippocampal tissues. Eighteen differentially expressed cognition-related proteins might be related to cognitive impairment in mice exposed to chronic high-altitude hypoxia.

Proteomics exploration of metformin hydrochloride for diabetic kidney disease treatment via the butanoate metabolism pathway

Diabetic nephropathy (DKD) is a diabetesrelated kidney injury with an increasing incidence every year. Metformin hydrochloride (MET), a cornerstone treatment for glucose lowering, has been widely reported for the treatment of DKD, but the specific molecular mechanisms and potential therapeutic targets still need to be further explored. We used kidney tissues from db/db mice as samples and used proteomics and bioinformatics to analyse the function, distribution and related pathways of differential proteins in DKD, focusing on the assessment of the binding energies of key proteins in the butyrate pathway and drugs at the molecular level, which showed that the expression profiles of differential proteins in kidney tissues were altered after MET treatment, involving energy metabolism. The key proteins involved in the butanoate metabolism pathway, including AACS, ACSM3, EHHADH and HMGCS2, exhibit binding energies to MET of ＜-5 kcal. It is therefore plausible that MET treatment may affect the butanoate metabolism pathway, potentially ameliorating the progression of DKD by modulating mitochondrial function and inflammatory responses.

GlycoPCT: Pressure Cycling Technology-Based Quantitative Glycoproteomics Reveals Distinctive N-Glycosylation in Human Liver Biopsy Samples of Nonalcoholic Fatty Liver Disease.

Protein N-glycosylation is vital in the human liver and influences functions such as lipid metabolism, apoptosis, and inflammation. However, site-specific N-glycosylation patterns and variations in liver biopsy samples between healthy individuals and those with nonalcoholic fatty liver disease (NAFLD) remain incompletely characterized, primarily due to the limitations of current clinical glycoproteomic methods, including a large demand for clinical samples, low efficiency of tissue protein extraction, and a low recovery rate of intact N-glycopeptides (IGPs). To address this issue, we developed GlycoPCT, a quantitative glycoproteomic method based on pressure cycling technology. It enables efficient recovery of IGPs and accurate analysis of trace liver biopsy samples. Our research revealed a total of 4,459 unique IGPs and 361 glycans from 758 glycoproteins. High-mannose type, complex type, fucosylation type, and sialylation type N-glycans were significantly upregulated in the NAFLD group (p < 0.001, t test). Notably, we also identified 182 upregulated IGPs from 67 proteins (p < 0.05, FC > 1.50) and 108 downregulated IGPs from 44 proteins (p < 0.05, FC < 0.67) in the NAFLD group. Furthermore, we highlighted an essential acute phase glycoprotein, alpha-1-acid glycoprotein 1 (A1TA), which is synthesized in the liver and plays a significant role in NAFLD progression. These novel glyco-signatures provide crucial clues for the diagnosis and pathogenesis of NAFLD.

Proteomic Analysis of Tissue Proteins Related to Lateral Lymph Node Metastasis in Papillary Thyroid Microcarcinoma.

Patients with lateral lymph node metastasis (LLNM) may experience higher locoregional recurrence rates and poorer prognoses compared to those without LLNM, highlighting the need for effective preoperative stratification to reliably assess risk LLNM. In this study, we collected PTMC samples from Peking Union Medical College Hospital and employed data-independent acquisition mass spectrometry proteomics technique to identify protein profiles in PTMC tissues with and without LLNM. Pseudo temporal analysis and single sample gene set enrichment analysis were conducted in combination with The Cancer Genome Atlas Thyroid Carcinoma for functional coordination analysis and the construction of a prediction model based on random forest. Non-negative matrix factorization (NMF) clustering was utilized to classify molecular subtypes of PTMC. Our findings revealed that the differential activation of pathways such as MAPK and PI3K was critical in enhancing the lateral lymph node metastatic potential of PTMC. We successfully screened biomarkers via machine learning and public databases, creating an effective prediction model for metastasis. Additionally, we explored the mechanism of metastasis-associated PTMC subtypes via NMF clustering. These insights into LLNM mechanisms in PTMC may contribute to future biomarker screening and the identification of therapeutic targets.

Deciphering the prognostic landscape of triple-negative breast cancer: A focus on immune-related hub genes and therapeutic implications.

Triple-negative breast cancer (TNBC), known for its hostile nature and limited treatment modalities, has spurred researchers to explore novel approaches for enhancing clinical outcomes. Here, the study aimed to analyze transcriptomics data to identify immune-related hub genes associated with TNBC that might serve as prognostic biomarkers. Initially, we determined genes that were differentially expressed between TNBC and normal tissues by integrating microarray and RNA sequencing data. Then, through protein-protein interaction and module analysis, we identified five putative hub genes: AURKA, CCNB1, CDCA8, GAPDH, and TOP2A. Subsequently, gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that the hub genes were primarily involved in the progesterone-mediated oocyte maturation signaling pathway and oocyte meiosis. Additionally, we observed that these five hub genes were significantly elevated at both protein and mRNA levels in TNBC tissues and contributed to worse survival. Furthermore, the expression of these hub genes exhibited a strong positive association with immune-invading cells such as CD8 T cells, CD4 T cells, and dendritic cells. The analysis of the regulatory network revealed three transcription factors (YBX-1, E2F1, and E2F3) and three posttranscriptional regulators (hsa-mir-25-3p, hsa-mir-92a-3p, and hsa-let-7b-5p) of hub genes. Finally, we explored potential drug candidates for the hub genes using Drug-Gene Interaction Database and discovered that there are no FDA-approved drugs for CCNB1 and CDCA8, highlighting a promising area for future research. Taken together, our results will be of immense importance in addressing the intricacies of TNBC.

Intranasal dantrolene nanoparticles inhibit inflammatory pyroptosis in 5XFAD mice brains.

This study investigates the effects of intranasal dantrolene nanoparticles on inflammation and programmed cell death by pyroptosis in 5XFAD Alzheimer's Disease (AD) mice. 5XFAD and wild type (WT) B6SJLF1/J mice were treated with intranasal dantrolene nanoparticles (5 mg/kg), daily, Monday to Friday, for 12 weeks continuously, starting at 9 months of age. Blood and brain were harvested at 13 months of age, one month after completion of 12 weeks intranasal dantrolene nanoparticle treatment. Blood biomarkers function of liver (Alanine transaminase, ALT), kidney (Creatinine), and thyroid (TSH: Thyroid-stimulating hormone) were measured using ELISA. The changes of whole brain tissue proteins on Ca 2+ release channels on membrane of endoplasmic reticulum (type 2 ryanodine and type 1 InsP3 receptors, RyR-2 and InsP3R-1), lipid peroxidation byproduct malondialdehyde (MDA)-modified proteins, 4-HNE, pyroptosis regulatory proteins (NLR family pyrin domain containing 3 (NLRP3), cleaved caspase-1, full length or N-terminal of Gasdermin D (GSDMD), cytotoxic (IL-1, IL-18, IL-6, TNF-a) and cytoprotective (IL-10) cytokines, astrogliosis (GFAP), microgliosis (IBA-1) and synapse proteins (PSD-95, Synapsin-1) were determined using immunoblotting. Body weights were monitored regularly. Intranasal dantrolene nanoparticles significantly inhibited the increase of RyR-2 and InsP3R-1 proteins, MDA-modified proteins, 4-NHE, pyroptosis regulatory proteins (NLRP3, cleaved caspase-1, N-terminal GSDMD), cytotoxic cytokine (IL-1β, IL-18, IL-6, TNF-α), biomarkers for astrogliosis (GFAP) and microgliosis (IBA-1), and the decrease of cytoprotective cytokine (IL-10) and synaptic proteins (PSD-95, synpasin-1). Intranasal dantrolene nanoparticles for 12 weeks did not affect blood biomarkers for function of liver, kidney, and thyroid, not did it change body weight significantly. Intranasal dantrolene nanoparticles significantly inhibit the increase of RyR-2 and InsP 3 R-1 Ca 2+ channel receptor proteins, ameliorate activation of the pyroptosis pathway and pathological inflammation, and the associated loss of synapse proteins. Intranasal dantrolene nanoparticles for three months did not affect liver, kidney and thyroid functions or cause other side effects.