Mitochondria plays an important regulatory role in the anticancer immune response by directly participating in the activation, proliferation and differentiation of immune cells, regulating multiple immune signaling pathways and affecting the activation and function of tumor-infiltrating immune cells with the help of the change of mitochondrial dynamics and metabolic. Mitochondria-targeted immunotherapy is an emerging cancer treatment strategy. The constructed mitochondrial targeted nanoformulation significantly improves the anti-cancer efficacy by inducing immunogenic cell death, regulating mitochondrial dynamics and metabolism, activating immune signaling pathways and other immune strategies. This review focuses on the field of cancer immunotherapy, introduces the relationship between mitochondria and tumor immunity, and summarizes the design of mitochondria-targeted nanomaterials in the past five years. Finally, the challenges and prospects of mitochondria-targeted nanoformulations in anti-cancer immunotherapy are discussed in this paper, which offers support for the development of novel mitochondria-targeted agents for cancer immunotherapy in the future.

It has become an industry consensus to take the concept of quality by design (QbD) as the basic principle of drug development design. On the basis of reviewing the design steps of pharmaceutical QbD, this paper deeply discusses the application of QbD in the development of lipid-based cubic liquid crystalline, and analyzes in detail the QbD elements in the design of lipid-based cubic liquid crystalline, such as quality target product profile, critical material attributes, risk assessment and factor screening, critical material attributes, critical process parameters, experimental design, design space and control strategy, etc. The purpose of this study is to deeply explore the integration and application of QbD concepts and provides a reference for the development of lipid-based cubic liquid crystalline preparations.

Dry granulation is a highly mechanized method for the preparation of drug granules. Roller compaction is a commonly used dry granulation method. The preparation of roller-compacted ribbons is a key step in dry granulation, and the properties of ribbons, such as density, solid fraction, and porosity, is related to the formation rate of drug granules, and directly determine their quality. The conventional method for measuring ribbons properties, which is off-line and time-consuming, can no longer meet the needs of continuous production. The emerging process analysis technology is capable of online real-time detection of ribbons properties. Combined with data analysis technology, it can provide feedback, control, and improve the ribbons preparation process to obtain high-quality ribbons, which is beneficial for continuous production of roller compaction. The method of detecting ribbon properties based on process analysis technology has made significant progress, and has become a hot topic in the analysis and control of roller compaction processes. This article systematically summarizes the latest progress in conventional determination methods and process analysis methods of ribbon properties, in order to provide theoretical guidance for the monitoring of ribbon properties and the continuous production of roller compaction dry granulation.

The intestinal mucus layer is mainly composed of mucin 2 (MUC2), a highly glycosylated protein secreted by goblet cells. The Muc2^‒/‒ mice are a spontaneous colitis model, due to intestinal mucus barrier defects, gut microbes directly contact with intestinal epithelial cells and induce intestinal inflammation. Nevertheless, how gut microbes affect the intestinal epithelium and trigger inflammation in the absence of mucus remains unknown. This study analyzed histopathological changes, colonic transcriptome and gut microbiome of Muc2^‒/‒ mice to elucidate the role of gut microbes in inducing intestinal inflammation. Animal experimentations were approved by the Institutional Animal Care and Use Committee of Nanjing University of Chinese Medicine, Ethical Review No. 202302A007. Pathological analysis showed that Muc2^‒/‒ mice manifested considerable intestinal mucosal damage accompanied by inflammatory cell infiltration and goblet cell exhaustion. Transcriptomic analysis showed that mucus synthesis, endoplasmic reticulum stress, and autophagy-related gene expressions were altered in Muc2^‒/‒ mice. Kyoto Encyclopedia of Genes and Gene Ontology pathway enrichment analyses showed considerable enrichment of inflammation and immune-related pathways, and pathways in response to bacteria derived molecules were upregulated. Microbial gene sequencing revealed dysbiosis of the gut microbiome in Muc2^‒/‒ mice, with Akkermansia muciniphila expansion being the characteristic profile. The expansion of Akkermansia muciniphila was confirmed in Il10^‒/‒ mice, which also displayed mucus barrier defects. Correlation analysis showed that the disturbed gut microbes of Muc2^‒/‒ mice was positively correlated with intestinal inflammation and immune-related differentially expressed genes, which were enriched in pathways such as MAPK cascade and chemokine production. In conclusion, in the absence of intestinal mucus layer, gut microbes can induce immune response by affecting the colonic gene transcription, leading to sustained secretion of inflammatory cytokines and intestinal inflammation. This study investigated the mechanism by which gut microbiota induces colonic inflammation in the absence of the intestinal mucus layer through integrated analysis of colonic transcriptomic profiles and gut microbiota dynamics. The findings aim to identify therapeutic targets for colitis treatment and drug discovery.

As the "power factory" of cells, mitochondria are important organelles for maintaining cellular homeostasis. The depletion of glucose and oxygen causes mitochondrial damage when ischemic stroke occurs, leading to energy metabolism disorders, reactive oxygen species (ROS) aggregation, which further aggravates ischemic nerve injury. Therefore, the aim of this study is to establish a rat middle cerebral artery occlusion/reperfusion (MCAO/R) model in vivo to investigate whether LIG can reduce ischemic nerve injury by restoring energy metabolism and the mechanism. The animal experiment protocol was approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine before implementation (animal ethics number: AHUCM-rats-2021030). TTC staining, neurological function score and brain index were used to evaluate nerve injury in MCAO/R rats. The levels of ROS, mitochondrial membrane potential, ATP, lactic acid and mitochondrial respiratory chain enzyme complex Ⅰ-Ⅳ were detected. The expression of glucose transporter 4 (GLUT 4) was detected by Western blot. Ligustilide was used for molecular docking with AMPK. Western blot was used to detect the protein expression levels of adenosine 5'-monophosphate (AMP)-activated protein kinase [AMPK], p-AMPK, and peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) in HT22 cells damaged by oxygen-glucose deprivation/reoxygenation (OGD/R). The results showed that LIG treatment significantly reduced cerebral infarct volume, brain index and neurobehavioral score in MCAO/R rats. It inhibited the aggregation of ROS, increased mitochondrial membrane potential and ATP content, and reduced lactic acid production. Molecular docking showed that LIG had a good binding ability with AMPK. In addition, LIG increased the protein expression of p-AMPK, AMPK and PGC-1α in HT22 cells damaged by OGD/R. These results suggest that LIG alleviates ischemic brain injury by improving mitochondrial function and restoring energy metabolism, and the mechanism may be related to the activation of AMPK/PGC-1α signaling pathway.

Drug targeting of the human ether-à-go-go-related gene (hERG)-encoded rapid delayed rectifier potassium channel induces acquired long QT syndrome (acLQTS), predisposing individuals to life-threatening arrhythmias. Previous studies demonstrate that the antipsychotic agent thioridazine (THIO) triggers acLQTS. Our findings aimed to elucidate the underlying mechanisms of the therapeutic effects against drug-induced cardiotoxicity. Using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) integrated with multi-electrode arrays (MEA) and optical mapping technology, we evaluated and compared the efficacy of tanshinone IIA (TANIIA), propranolol (Prop) and resveratrol (Res) in reversing THIO-induced prolongation of action potential duration (APD) and field potential duration (all animal experiments were conducted in accordance with the Animal Care Guidelines of Harbin Medical University and were approved by the Ethics Committee of Harbin Medical University, the approval number is IRB3020622). The results showed that TANIIA significantly reversed the THIO-induced prolongation of APD₉₀, APD₅₀, and field potential, whereas neither Prop nor Res counteracted this effect. Building on these results, we further explored the mechanistic basis and molecular targets underlying TANIIA's reversal effects. Western blot and whole-cell patch-clamp demonstrated that TANIIA restored hERG protein expression downregulated by THIO and rescued its channel trafficking defects and rescued channel trafficking defects. Mechanistically, TANIIA attenuated THIO-induced aberrant interactions between hERG and the calnexin, thereby rectifying impaired channel folding and trafficking. Concurrently, TANIIA substantially reduced THIO-induced L-type calcium current. In conclusion, TANIIA ameliorates THIO-induced acLQTS by up regulating hERG current expression and down regulating L-type calcium current. These findings highlight its therapeutic potential of drug-induced acLQTS for clinical.

Ribozymes and DNAzymes are limited in their applications as nucleic acid therapeutics due to their strong dependence on metal ions. Therefore, the development of novel metal-independent ribozymes has become a key research focus. This study evaluated the fidelity of wild-type Tgo DNA polymerase in simultaneously incorporating sugar backbone-modified 2'-deoxy-2'-fluoroarabinonucleic acid (FANA) along with base-modified DNA, constructed a modified nucleotide library containing four modified nucleotides, and employed systematic evolution of ligands by exponential enrichment (SELEX) technology to select ribozymes with RNA cleavage activity. Unlike conventional methods for selecting modified functional nucleic acids, the modified library here was directly used as templates for PCR amplification, eliminating the reverse transcription step which copies the modified phenotypic functional sequence into unmodified cDNA, and dramatically simplified the screening process. This study employed an RNA substrate region composed of 15 consecutive ribonucleosides for selection. Through sequence homology analysis and secondary structure prediction, we verified the cleavage activity of the top twenty abundant sequences obtained from sequencing the most active round 4 library. A sequence denoted R4-11 with considerably high RNA cleavage activity was successfully identified, and it preferentially cleaves the phosphodiester linkage between the rC-rA dinucleotide. This study provides a method for selecting ribozymes composed of diverse modified nucleotides and offers novel tools for disease-targeted therapy through mRNA silencing.

Candida albicans (C. albicans) is the most common opportunistic pathogen in clinical fungal infections, and its invasive pathogenic mechanisms and drug resistance are closely associated with biofilm formation. Therefore, research on biofilm inhibitors holds significant importance. This study found that N-(2,4-dimethylphenyl) maleimide (abbreviated as IMB-D7) can inhibit the formation of C. albicans biofilms and has a certain clearance effect on mature biofilms. IMB-D7 significantly reduces the adhesion capacity and hydrophobicity of C. albicans cells, inhibits the yeast-hyphae phase transition and hyphae formation, and affects the content of key components of the cell wall. IMB-D7 significantly downregulates the expression of genes associated with adhesion, hyphal formation, and cell wall component synthesis. Additionally, IMB-D7 exhibits low cytotoxicity toward human oral epithelial cells. Therefore, further research on IMB-D7 holds promise for the discovery of novel antifungal active lead compounds.

This study systematically investigated the therapeutic mechanism of matrine derivative, (6aS, 10S, 11aR, 11bR, 11cS)-10-methylaminododecahydro-3a, 7a-diaza-benzo (de) anthracene-8-thione (MASM), in experimental autoimmune encephalomyelitis (EAE) through multi-dimensional analysis. The myelin oligodendrocyte glycoprotein MOG_35-55/pertussis toxin-induced EAE mouse model was established, and neurological impairment was confirmed using Kono's 5-point scoring system, mice with scores between 0.5 and 3.5 were considered successful models. These mice were randomly divided into six groups: EAE control, MASM groups (1.5, 3 and 6 mg·kg^-1), a positive control group (fingolimod, 1.0 mg·kg^-1), and a normal control group. Drug administration was initiated on the day of grouping, once daily for 21 consecutive days. The Kono's score was used to evaluate the neurological function; HE staining and electron microscopy was used to observe the pathological changes of spinal cord; transforming growth factor-β1 and monocyte chemoattractant protein-1 content was detected by ELISA. The gene expression of H2T23 was detected by qRT-PCR, the binding interactions between MASM and human frizzled homolog (FZD5) was evaluated by molecular docking, and protein expression levels of glial fibrillary acidic protein (GFAP), complement 3 (C3), wingless type mouse mammary tumor virus integration site family member 5a (Wnt5a), FZD5, protein kinase Cα (PKCα), and the phosphorylation level of glycogen synthase kinase 3 (GSK3β) were detected by Western blot. MASM significantly improved neurological function, reduced spinal cord inflammation and demyelination, and downregulated astrocyte activation markers (GFAP, C3, H2T23) (P < 0.05, P < 0.01) and Wnt5a/FZD5 pathway components (Wnt5a, FZD5, PKCα, p-GSK3β) (P < 0.05, P < 0.01). MASM demonstrates strong binding capacity to FZD. These findings suggest that MASM alleviates EAE by inhibiting astrocyte polarization via the Wnt5a/FZD5 pathway. The animal study was approved by the Animal Ethics Committee of Fujian University of Traditional Chinese Medicine (approval number: FJTCMIACUC 2023319).

M1 macrophages play a critical role in tumorigenesis and development. Insufficient infiltration of M1 macrophages can lead to inadequate CD8⁺ T cell activation, impairing immune surveillance and thereby promoting immune escape. This study investigated the mechanism by which baicalin induces the polarization of RAW264.7 cells towards the M1 phenotype using an in vitro cell model. Cell viability was assessed by the CCK-8 assay. The expression of the M1 surface marker CD86 was analyzed by flow cytometry and immunofluorescence. Cytokine tumor necrosis factor-alpha (TNF-α) levels were measured by ELISA. Lactate levels were detected using a lactate assay kit. Western blot was used to determine the protein expression of key components of the PI3K/Akt/HIF-1α glycolytic pathway and the key rate-limiting glycolytic enzymes lactate dehydrogenase A (LDHA) and pyruvate kinase M2 (PKM2) in RAW264.7 cells. The PI3K inhibitor LY294002 and the glycolytic inhibitor 2-DG were employed to validate the mechanism of action of baicalin. The results showed that baicalin at concentrations of 12.5-100 μmol·L^-1 had no significant effect on the viability of RAW264.7 cells. At 100 μmol·L^-1, baicalin significantly up-regulated the expression of the M1 surface marker CD86 and the cytokine TNF-α, increased the protein expression of p-PI3K, p-Akt, HIF-1α, LDHA, and PKM2, and elevated lactate levels. However, the up-regulatory effects of baicalin on CD86 were inhibited, and the expression of related pathway proteins was reversed, upon addition of LY294002 or 2-DG. This study reveals for the first time that baicalin drives macrophage polarization towards the M1 phenotype by activating the PI3K/Akt/HIF-1α signaling axis and enhancing glycolytic metabolism. This finding provides new mechanistic evidence based on metabolism for its therapeutic potential in tumor immune regulation and inflammation-related diseases.