Molecular Mechanisms Research Articles

Endoplasmic reticulum stress in acute lung injury and pulmonary fibrosis.

Pulmonary fibrosis (PF) is a progressive and irreversible lung disease that leads to diminished lung function, respiratory failure, and ultimately death and typically has a poor prognosis, with an average survival time of 2 to 5 years. Related articles suggested that endoplasmic reticulum (ER) stress played a critical role in the occurrence and progression of PF. The ER is responsible for maintaining protein homeostasis. However, factors such as aging, hypoxia, oxidative stress, or inflammation can disrupt this balance, promoting the accumulation of misfolded proteins in the ER and triggering ER stress. To cope with this situation, cells activate the unfolded protein response (UPR). Since acute lung injury (ALI) is one of the key onset events of PF, in this review, we will discuss the role of ER stress in ALI and PF by activating multiple signaling pathways and molecular mechanisms that affect the function and behavior of different cell types, with a focus on epithelial cells, fibroblasts, and macrophages. Linking ER stress to these cell types may broaden our understanding of the mechanisms underlying lung fibrosis and help us target these cells through these mechanisms. The relationship between ER stress and PF is still evolving, and future research will explore new strategies to regulate UPR pathways, providing novel therapeutic targets.

Towards the Albino Mutant Gene in Malus × Domestica Borkh.

Albino mutation is among the most common phenomena that often causes a water imbalance and disturbs physiological functions in higher species of trees. Albinism frequently occurs in hybridized apples, but almost all seedlings die shortly after germination. In this study, a spontaneous albino mutant on Fuji apple trees was obtained. After bud grafting, new albino shoots with greenish-white leaves grew, although they were slender, small, and died easily. Resequencing analysis indicated that a total of 49.37 Gbp clean data of the albino mutant samples was obtained; its Q30 reached 91.43%, the average rate mapped was 93.69%, and genome coverage was 96.47% (at least one base cover). Comparisons of the sequences for the albino mutants revealed 4,817,412 single-nucleotide polymorphisms (SNPs), 721,688 insertion/deletion markers (InDels), and 43,072 structural variations (SVs). The genes with non-synonymous SNPs, InDels, and SVs in CDS were compared with KEGG, GO, COG, NR, and SwissProt databases, and a total of 5700 variant genes were identified. A total of 1377 mutant genes had the GO annotation information. Among these, 1520 mutant genes had the pathway annotation and took part in 123 pathways. A total of 1935 variant genes were functionally classified into 25 COG categories. Further research on these variants could help understand the molecular regulatory mechanism of the apple albino mutant. Similarly, variations in the homologous MdAPG1 (Albino or pale-green mutant 1) gene, which was located on Chromosome 11 and belonged to the S-adenosyl-L-methionine-dependent methyltransferases superfamily, may have led to the generation of this apple albino mutant.

Gene recognition and role of foodomics in mycotoxin control: A review

Since recognition of toxic and carcinogenic aflatoxins in Brazilian groundnut meal in 1960, much research has been done to prevent and detoxify aflatoxins in foods and feeds, identifying a variety of methods. The research has expanded to other mycotoxins. The biotic and abiotic factors favoring mycotoxin contaminations have been understood through experiments under laboratory conditions and analysis of field data. However, many gaps remain in the knowledge on mycotoxin control at the molecular level that may be useful in addressing mycotoxigenic hazards. Recognition of responsible genes in hosts and fungi and omics methods applying genomics, transcriptomics, proteomics, and metabolomics to understand mycotoxin biosynthesis at the molecular level may open new avenues to interact with plant-fungi-bacteria cross-talks, apply regulatory mechanisms in biosynthesis, and explore checks and controls addressing abiotic and biotic factors favoring mycotoxin biosynthesis. The new knowledge is expected to generate probable molecular biological mechanisms to eliminate mycotoxin biosynthesis on foods. The current level of omics knowledge requires application of research to achieve deeper understanding, aiming at new methods for mycotoxin controls and applying next-generation technologies. This review examines the current knowledge on the biosynthesis of aflatoxins, fusarium toxins, and patulin in foods and host-fungi interactions at a molecular level.

Beneficial microbiome and diet interplay in early-onset colorectal cancer.

Colorectal cancer (CRC) is the third most commonly diagnosed cancer and the second leading cause of cancer-related deaths worldwide. Although the risk of developing CRC increases with age, approximately 10% of newly diagnosed cases occur in individuals under the age of 50. Significant changes in dietary habits in young adults since industrialization create a favorable microenvironment for colorectal carcinogenesis. We aim here to shed light on the complex interplay between diet and gut microbiome in the pathogenesis and prevention of early-onset CRC (EO-CRC). We provide an overview of dietary risk factors associated with EO-CRC and contrast them with the general trends for CRC. We delve into gut bacteria, fungi, and phages with potential benefits against CRC and discuss the underlying molecular mechanisms. Furthermore, based on recent findings from human studies, we offer insights into how dietary modifications could potentially enhance gut microbiome composition to mitigate CRC risk. All together, we outline the current research landscape in this area and propose directions for future investigations that could pave the way for novel preventive and therapeutic strategies.

OCTN1 (SLC22A4) as a Target of Heavy Metals: Its Possible Role in Microplastic Threats

Microplastics represent a threat due to their ability to enter the food chain, with harmful consequences for living organisms. The riskiness of these particles is also linked to the release of other contaminants, such as heavy metals. Solute Carriers (SLCs) represent eminent examples of first-level targets of heavy metals due to their localization on the cell surface. Putative targets of heavy metals are the organic cation transporters that form a sub-clade of the SLC22 family. Besides the physiological role in the absorption/release of endogenous organic cations, these transporters are crucial in drug disposition and their interaction with xenobiotics. In this work, the human SLC22A4, commonly known as OCTN1, was used as a benchmark to test interactions with heavy metals released by microplastics, exploiting the proteoliposome tool. The potency of metals to interfere with the OCTN1 function has been evaluated by measuring IC50 values calculated in the micromolar range. The molecular mechanism of interaction has been defined using site-directed mutagenesis and computational analyses. Finally, some chemical and physiological thiol-reacting compounds show the capacity to rescue the metal-inhibited OCTN1 function. The conclusions drawn on OCTN1 can be extended to other members of the SLC22 family and orthologous transporters in fish.

Unified bursting strategies in ectopic and endogenous even-skipped expression patterns

Transcription often occurs in bursts as gene promoters switch stochastically between active and inactive states. Enhancers can dictate transcriptional activity in animal development through the modulation of burst frequency, duration, or amplitude. Previous studies observed that different enhancers can achieve a wide range of transcriptional outputs through the same strategies of bursting control. For example, in Berrocal et al., 2020, we showed that despite responding to different transcription factors, all even-skipped enhancers increase transcription by upregulating burst frequency and amplitude while burst duration remains largely constant. These shared bursting strategies suggest that a unified molecular mechanism constraints how enhancers modulate transcriptional output. Alternatively, different enhancers could have converged on the same bursting control strategy because of natural selection favoring one of these particular strategies. To distinguish between these two scenarios, we compared transcriptional bursting between endogenous and ectopic gene expression patterns. Because enhancers act under different regulatory inputs in ectopic patterns, dissimilar bursting control strategies between endogenous and ectopic patterns would suggest that enhancers adapted their bursting strategies to their trans-regulatory environment. Here, we generated ectopic even-skipped transcription patterns in fruit fly embryos and discovered that bursting strategies remain consistent in endogenous and ectopic even-skipped expression. These results provide evidence for a unified molecular mechanism shaping even-skipped bursting strategies and serve as a starting point to uncover the realm of strategies employed by other enhancers.

Potentials of Mean Force and Solvent Effects of the CN- + CH3X (X = F, Cl, Br, and I) Reactions by the N-Side Attack in Aqueous Solution.

A combined multilevel quantum mechanics and molecular mechanics approach is performed to investigate the nucleophilic substitution reactions of CN- + CH3X (X = F, Cl, Br, and I) by the N-side attack in aqueous solution. The water molecules are treated microscopically using an explicit SPC/E model, and the potentials of mean force are characterized by both the DFT and CCSD(T) levels of theory for the solute. Calculations demonstrate that the shielding effect of the solvent reduces the nucleophile-substrate and substrate-leaving group interactions in solution, leading to stationary point structures that are quite different from those in the gas phase. The structure and charge evolution along the reaction paths reveal that the reaction is not only a synchronous bonding and bond-breaking Walden-inversion mechanism but also a synchronous charge transfer process. The activation barriers calculated at the CCSD(T) level of theory are 27.5 (F), 22.6 (Cl), 21.7 (Br), and 21.2 (I) kcal/mol, respectively, which are larger than the corresponding experimental values for the C-side attack. The polarization effect of water molecules causing solute polarization contributes to the activation barrier in the order of F > Cl > Br > I. The solvent energy contribution to the activation barrier is in the order of F < Cl < Br < I because the F leaving group has the most compact transition state structure and the I leaving group has the loosest transition state structure. As a result, the total contributions of the solvent effects to the activation barriers are 7.9 (F), 10.7 (Cl), 15.3 (Br), and 15.7 (I) kcal/mol. Our results show that the solvent effects have a significant influence on both the structure and the energetics of the N-side attack reactions in aqueous solution.

Comprehensive analysis of IRF8-related genes and immune characteristics in lupus nephritis

BackgroundThere are currently no reliable diagnostic biomarkers or treatments for lupus nephritis (LN), a complication of systemic lupus erythematosus. Objective: We aimed to explore gene networks and potential biomarkers for LN by analyzing the GSE32591 and GSE113342 datasets from the Gene Expression Omnibus database, focusing on IRF8 and IRF8-related genes.MethodsWe used differential expression analysis, functional enrichment, protein-protein interaction (PPI) network construction, and the CIBERSORT algorithm for immune infiltration assessment. To validate the expression levels of the IRF8 gene in the kidneys of lupus mice models, we used quantitative real-time PCR (qRT-PCR) and Western blotting (WB). A diagnostic classifier was built using the RandomForest method to evaluate the diagnostic potential of selected key genes. To bridge our findings with potential therapeutic implications, we used the drug-gene interaction database to predict drugs targeting the identified genes.ResultsTwenty co-differentially expressed genes (DEGs) were identified, with IRF8 exhibiting significant expression differences and potential as a biomarker. Functional enrichment analysis revealed pathways associated with immune response. Validation through qRT-PCR and WB confirmed that the IRF8 gene and its protein exhibited elevated expression levels in the kidneys of lupus mice compared to control groups. The diagnostic classifier revealed impressive accuracy in differentiating LN from control samples, achieving a notable area under the curve values across various datasets. Additionally, immune infiltration analysis indicated significant differences in the immune cell profiles between the LN and control groups.ConclusionIRF8 and its related genes show promise as biomarkers and therapeutic targets for LN. These findings contribute to a deeper understanding of the molecular mechanisms involved in LN and may support the development of precision medicine strategies for improved patient outcomes.

Molecular basis of resistance to leaf spot disease in oil palm

IntroductionLeaf spot disease caused by the fungal pathogen Curvularia oryzae is one of the most common diseases found in oil palm (Elaeis guineensis) nurseries in South East Asia, and is most prevalent at the seedling stage. Severe infections result in localized necrotic regions of leaves that rapidly spread within nurseries leading to poor quality seedlings and high economic losses.MethodsTo understand the molecular mechanisms of this plant-pathogen interaction, RNA-Seq was used to elucidate the transcriptomes of three oil palm genotypes with contrasting pathogen responses (G10 and G12, resistant and G14, susceptible) following infection with C. oryzae spores. Transcriptomes were obtained from Illumina NovaSeq 6000 sequencing of mRNA at four different time points (day 0, before treatment; day 1, 7, and 21 post treatment).Results and discussionAnalysis of differentially expressed gene (DEG) profiles in these three genotypes provided an overview of the genes involved in the plant defence. Genes involved in disease resistance, phytohormone biosynthesis, gene regulation (transcription factors), and those encoding proteins associated with cell wall hardening were identified and likely contribute to the resistance of oil palm to C. oryzae. Such genes represent good candidates for targets to enhance oil palm productivity and resilience through molecular breeding approaches.

Fate and Effects of Heavy Metals in Fishes: Antioxidant Defense System, miRNA/Gene Expression Response, and Histopathological Reproductive Manifestations.

Heavy metal pollution is a major environmental concern and in particular for aquatic ecosystems. With heavy metals exceeding safe and recommended limits, they pose significant threats to the environment and its inhabitants, including fish. Heavy metals, when accumulated in the different organs of the fish, result in toxicity in fish by producing reactive oxygen species (ROS) through the generation of oxidizing radicals. This oxidative stress mechanism is a key factor in the detrimental effects of heavy metal pollution on aquatic life. Heavy metal exposure profoundly affects fish behavior and physiology. In this review, an attempt was made to report the effects of heavy metals on fish physiology, focusing on toxicological effects on antioxidant enzymes, microRNAs (miRNAs) and molecular genetic responses, histopathology of organs, and underlying molecular mechanisms. This review also highlighted the heavy metal impact on fish gonads (testes and ovaries) and the hormones associated with it. The detection methods and the incorporation of latest developments in AI-based technology for the detection of heavy metals are also included in this review. Understanding the above effects is important for assessing the ecological impact of heavy metal pollution and developing strategies to mitigate its adverse effects on aquatic life. Understanding the consequences listed above is important for analyzing the ecological impact of heavy metal pollution and devising measures to reduce its negative effects on aquatic life and human health.

Unified bursting strategies in ectopic and endogenous even-skipped expression patterns.

Transcription often occurs in bursts as gene promoters switch stochastically between active and inactive states. Enhancers can dictate transcriptional activity in animal development through the modulation of burst frequency, duration, or amplitude. Previous studies observed that different enhancers can achieve a wide range of transcriptional outputs through the same strategies of bursting control. For example, in Berrocal et al., 2020, we showed that despite responding to different transcription factors, all even-skipped enhancers increase transcription by upregulating burst frequency and amplitude while burst duration remains largely constant. These shared bursting strategies suggest that a unified molecular mechanism constraints how enhancers modulate transcriptional output. Alternatively, different enhancers could have converged on the same bursting control strategy because of natural selection favoring one of these particular strategies. To distinguish between these two scenarios, we compared transcriptional bursting between endogenous and ectopic gene expression patterns. Because enhancers act under different regulatory inputs in ectopic patterns, dissimilar bursting control strategies between endogenous and ectopic patterns would suggest that enhancers adapted their bursting strategies to their trans-regulatory environment. Here, we generated ectopic even-skipped transcription patterns in fruit fly embryos and discovered that bursting strategies remain consistent in endogenous and ectopic even-skipped expression. These results provide evidence for a unified molecular mechanism shaping even-skipped bursting strategies and serve as a starting point to uncover the realm of strategies employed by other enhancers.

Update on NKCC2 regulation in the thick ascending limb (TAL) by membrane trafficking, phosphorylation, and protein-protein interactions

Purpose of reviewThe thick ascending limb (TAL) of loop of Henle is essential for NaCl, calcium and magnesium homeostasis, pH balance and for urine concentration. NKCC2 is the main transporter for NaCl reabsorption in the TAL and its regulation is very complex. There have been recent advancements toward understanding how NKCC2 is regulated by protein trafficking, protein-protein interaction, and phosphorylation/dephosphorylation. Here, we update the latest molecular mechanisms and players that control NKCC2 function, which gives an increasingly complex picture of NKKC2 regulation in the apical membrane of the TAL.Recent FindingsProtein-protein interactions are required as a regulatory mechanism in many cellular processes. A handful of proteins have been recently identified as an interacting partner of NKCC2, which play major roles in regulating NKCC2 trafficking and activity. New players in NKCC2 internalization and trafficking have been identified. NKCC2 activity is also regulated by kinases and phosphatases, and there have been developments in that area as well.SummaryHere we review the current understanding of apical trafficking of NKCC2 in the thick ascending limb (TAL) which is tightly controlled by protein-protein interactions, protein turnover and by phosphorylation and dephosphorylation. We discuss new proteins and processes that regulate NKCC2 that have physiological and pathological significance.

Growth and physiological response of Yulu Hippophae rhamnoides to drought stress and its omics analysis

ABSTRACT Hippophae rhamnoides (H. rhamnoides) is the primary tree species known for its ecological and economic benefits in arid and semi-arid regions. Understanding the response of H. rhamnoides roots to drought stress is essential for promoting the development of varieties. One-year-old Yulu H. rhamnoides was utilized as the experimental material, and three water gradients were established: control (CK), moderate (T1) and severe (T2), over a period of 120 days. The phenotypic traits and physiological indies were assessed and analyzed, while the roots were subjected by RNA-Seq transcriptome and Tandem Mass Tags (TMT) proteome analysis. Drought stress significantly reduced the plant height, ground diameter, root biomass and superoxide dismutase activity; however, the main root length increased. In comparison with CK, a total of 5789 and 5594 differential genes, as well as 63 and 1012 differential proteins, were identified in T1 and T2, respectively. The combined analysis of transcriptome and proteome showed that the number of differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) associated with T1, T2 and CK was 28 and 126, respectively, with 7 and 36 genes achieving effective KEGG annotation. In T1 and T2, the differential genes were significantly enriched in the plant hormone signal transduction pathway, but there was no significant enrichment in the protein expression profile. In T2, 38 plant hormone signal transduction function genes and 10 peroxisome related genes were identified. With the increase of drought stress, the combined expression of DEGs and DEPs increased. Yulu H. rhamnoides may allocate more resources toward CAT while simultaneously decreasing SOD and POD to mitigate the oxidative stress induced by drought. Furthermore, the molecular mechanisms underlying plant hormone signal transduction and peroxisome-related genes in the roots of H. rhamnoides were discussed in greater detail.

Neuroprotective Effects of STAT3 Inhibitor on Hydrogen Peroxide-Induced Neuronal Cell Death via the ERK/CREB Signaling Pathway

This study investigates the neuroprotective potential of STAT3 inhibition in reducing oxidative stress-induced neuronal damage and apoptosis, a major factor contributing to the onset and progression of neurodegenerative diseases, including Alzheimer’s disease (AD). Our findings demonstrate that STAT3 inhibitors significantly enhance cell survival and reduce apoptosis in SH-SY5Y cells exposed to hydrogen peroxide. These protective effects are mediated through the ERK/CREB signaling pathway rather than direct suppression of STAT3 phosphorylation. Further analysis revealed that the ERK pathway is a critical mediator of CREB activation following STAT3 inhibition. The protective effects of STAT3 inhibitors were significantly reduced in the presence of the ERK inhibitor PD98059, underscoring the importance of the ERK/CREB axis in neuroprotection. We observed that STAT3 inhibitors promote CREB phosphorylation, leading to the upregulation of immediate early genes such as c-Fos, c-Jun, Arc, Egr-1, NR4A1, and Homer1a, as well as BDNF. These genes are crucial for synaptic plasticity and long-term memory formation, suggesting that STAT3 inhibition may ameliorate cognitive impairments in neurodegenerative conditions. Our results highlight the potential of STAT3 inhibitors to counteract oxidative stress and enhance cognitive functions by modulating the ERK/CREB signaling pathway. These findings provide valuable insights into the molecular mechanisms of STAT3 inhibition and support its therapeutic potential for treating neurodegenerative diseases.

Signaling pathways mediating the induction of preharvest fruit drop in litchi

Certain litchi varieties, such as “Nuomici”, are highly susceptible to preharvest fruit drop, which leads to significant losses in fruit yield and economic value. However, the precise molecular mechanisms underlying this issue are not yet fully understood. In this study, we aimed to elucidate the signaling pathways that facilitate preharvest fruit drop in litchi, using “Nuomici” and “Huaizhi” cultivars as examples, which demonstrate high and low preharvest fruit drop rates, respectively. Our findings revealed that “Nuomici” experienced a substantial preharvest fruit drop, with a cumulative rate of 41.68%, significantly higher than the 1.44% observed in “Huaizhi”. Cellulase activity assays showed a significant increase in cellulase activity in the abscission zone of “Nuomici”, which coincided with the occurrence of preharvest fruit drop, in contrast to the relatively low levels in “Huaizhi”. Phytohormone assays indicated lower indole-3-acetic acid content in the pericarp, aril, and seeds of “Nuomici” during the preharvest stage compared to “Huaizhi”, coupled with higher abscisic acid levels in the seeds of “Nuomici”. Furthermore, transcriptomic analysis identified 180, 282, 655, and 241 differentially expressed genes (DEGs) in the pericarp, aril, seed, and abscission zone, respectively, between the two cultivars during preharvest fruit drop. These DEGs are intricately involved in the generation and transmission of abscission signals from fruit tissues, encompassing PIN, PIN-LIKES, LAX, and SAUR genes related to polar auxin transport, ethylene diffusion, as well as perceiving these signals and activating the abscission process within the abscission zone. This includes ACO and ILR genes involved in hormone biosynthesis and signal transduction, regulation by WRKY, NAC, and bHLH transcription factors, AAO genes involved in response to reactive oxygen species, and EXP, EG, and PG genes involved in cell wall degradation in the abscission zone. Based on these comprehensive findings, we propose a model for preharvest fruit drop triggered by a series of molecular events in litchi, providing valuable insights into the complex mechanisms governing this phenomenon.

Molecular Insights into the Penetration Enhancement Mechanism of Terpenes to Skin.

Terpenes are widely used in cosmetic formulations as chemical penetration enhancers (CPEs). However, the molecular mechanisms underlying the skin-penetration-enhancing effect have not been completely understood. In this work, molecular dynamics (MD) simulations were used to explore the influence of terpineol (TER), 1,8-cineole (CIN), and limonene (LIM) on the stratum corneum (SC) model. The results indicated that terpenes affected lipid membranes in a concentration-dependent manner and promoted skin permeation by disorganizing the cholesterol (CHOL) portion. The penetration of CPEs across the skin membrane also differed, with diffusion rates of limonene > 1,8-cineole > terpineol. The limonene molecules could penetrate the bilayer's center, forming a "triple layer" membrane structure. Furthermore, constrained simulations showed that the most favorable penetration pathway is via areas rich in CHOL. Terpineol could lower the energy barrier of the hydrophilic molecule (caffeic acid) across the cholesterol region. For the lipophilic molecule (osthole), limonene and 1,8-cineole could reduce the energy barrier across the cholesterol region. Each of the results provides novel insights into the mechanism of penetration of CPEs in the skin.

Transcriptome Analysis Reveals Sertoli Cells Adapting Through Redox and Metabolic Pathways Under Heat Stress in Goats

Background/Objectives: Climate change-induced temperature elevations pose significant challenges to livestock reproduction, particularly affecting testicular function in small ruminants. This study investigates the acute heat-stress response in goat Sertoli cells (SCs), aiming to elucidate the molecular mechanisms underlying heat-induced damage to male reproductive tissues. Methods: SCs were isolated from testes of 4-month-old black goats and exposed to heat stress (44 °C for 2.5 h). We employed transcriptome sequencing, CCK-8 assay, electron microscopy, ROS measurement, autophagy detection, Western blot analysis, and lactate concentration measurement. Bioinformatics analyses including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and protein–protein interaction network analyses were performed on the transcriptome data. Results: Heat stress significantly reduced SC viability, induced oxidative stress and autophagy, and altered gene expression profiles. We identified 1231 significantly differentially expressed genes, with significant enrichment in membrane-related processes and metabolic pathways. Metabolism-related genes, including PKLR, ACOT11, and LPCT12, were significantly downregulated. Protein–protein interaction network analysis revealed ten hub genes potentially crucial in the heat-stress response: HSP90AA1, HSPA5, BAG3, IGF1, HSPH1, IL1A, CCL2, CXCL10, ALB, and CALML4. Conclusions: This study provides comprehensive insights into the molecular mechanisms underlying goat SC response to heat stress. The identified genes and pathways, particularly those related to metabolism and stress response, offer potential targets for developing strategies to mitigate heat-stress effects on livestock reproduction. These findings contribute to our understanding of climate change impacts on animal husbandry and may inform the development of heat-stress resistant livestock lines.

2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione mediates the effect of ROS-enhanced PI3K/Akt/mTOR pathway on autophagy in breast cancer.

Several studies have suggested a potential antitumor effect of 2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione (DMDD). To further understand the mechanism of action of this compound, we investigated its effect on the phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. We show that DMDD application significantly inhibited the proliferation of breast cancer cell lines MDA-MB-231 and ER-α positive MCF-7. Furthermore, DMDD application resulted in increased intracellular reactive oxygen species (ROS) levels, apoptosis and autophagy, whereas it downregulated the expression of PI3K, Akt and mTOR mRNA and proteins, and increased the expression of LC3II/I and p62 proteins. In a mouse breast cancer xenograft model, DMDD inhibited tumor growth. Expression analyses suggest that ROS levels were higher in DMDD treated tumor tissues, whereas immunohistochemical analyses suggest that apoptotic cells were more prevalent in the DMDD treated group compared to the control group. Taken together, our results suggest that the molecular mechanism of action of DMDD may involve the enhancement of breast cancer autophagy through the PI3K/Akt/mTOR signaling pathway by mediating ROS expression.

Innovative strategies to optimise colorectal cancer immunotherapy through molecular mechanism insights

BackgroundColorectal cancer (CRC) is a leading cause of cancer-related deaths globally. The heterogeneity of the tumor microenvironment significantly influences patient prognosis, while the diversity of tumor cells shapes its unique characteristics. A comprehensive analysis of the molecular profile of tumor cells is crucial for identifying novel molecular targets for drug sensitivity analysis and for uncovering the pathophysiological mechanisms underlying CRC.MethodsWe utilized single-cell RNA sequencing technology to analyze 13 tissue samples from 4 CRC patients, identifying key cell types within the tumor microenvironment. Intercellular communication was assessed using CellChat, and a risk score model was developed based on eight prognostic genes to enhance patient stratification for immunotherapeutic approaches. Additionally, in vitro experiments were performed on DLX2, a gene strongly associated with poor prognosis, to validate its potential role as a therapeutic target in CRC progression.ResultsEight major cell types were identified across the tissue samples. Within the tumor cell population, seven distinct subtypes were recognized, with the C0 FXYD5+ tumor cells subtype being significantly linked to cancer progression and poor prognosis. CellChat analysis indicated extensive communication among tumor cells, fibroblasts, and immune cells, underscoring the complexity of the tumor microenvironment. The risk score model demonstrated high accuracy in predicting 1-, 3-, and 5-year survival rates in CRC patients. Enrichment analysis revealed that the C0 FXYD5+ tumor cell subtype exhibited increased energy metabolism, protein synthesis, and oxidative phosphorylation, contributing to its aggressive behavior. In vitro experiments confirmed DLX2 as a critical gene associated with poor prognosis, suggesting its viability as a target for improving drug sensitivity.ConclusionIn summary, this study advances our understanding of CRC progression by identifying critical tumor subtypes, molecular pathways, and prognostic markers that can inform innovative strategies for predicting and enhancing drug sensitivity. These findings hold promise for optimizing immunotherapeutic approaches and developing new targeted therapies, ultimately aiming to improve patient outcomes in CRC.

A novel anti-epileptogenesis strategy of temporal lobe epilepsy based on nitric oxide donor.

The molecular mechanism underlying the role of hippocampal hilar interneuron degeneration in temporal lobe epilepsy (TLE) remains unclear. Especially, very few studies have focused on the role of neuronal nitric oxide synthase (nNOS, encoded by Nos1) containing hilar interneurons in TLE. In the present study, Nos1 conditional knockout mice were constructed, and we found that selective deletion of Nos1 in hilar interneurons rather than dentate granular cells (DGCs) triggered epileptogenesis. The level of nNOS was downregulated in patients and mice with TLE. Nos1 deletion led to excessive epilepsy-like excitatory input circuit formation and hyperexcitation of DGCs. Replenishment of hilar nNOS protein blocked epileptogenic development and memory impairment in pilocarpine-induced TLE mice. Moreover, chronic treatment with DETA/NONOate, a slowly released exogenous nitric oxide (NO) donor, prevented aberrant neural circuits of DGCs and the consequent epileptogenesis without acute antiseizure effects. Therefore, we concluded that NO donor therapy may be a novel anti-epileptogenesis strategy, different from existing antiseizure medications (ASMs), for curing TLE.