Oxaliplatin (Oxa) is a chemotherapy drug commonly used for advanced colorectal cancer, however most patients develop resistance after treatment while the mechanisms of which have not been fully elucidated. In this study, oxaliplatin resistant cell lines were constructed from human colorectal cancer HCT116 cells through concentration gradient induction. On this basis, we investigated the expression profiling of HCT116/Oxa cells based on quantitative proteomics. Gene ontology (GO) analysis was conducted via The Database for Annotation, Visualization, and Integrated Discovery Database (DAVID), and pathway enrichment analysis was done using GeneAnalytics database. The potential targets and molecular mechanisms of oxaliplatin resistance in colorectal cancer were further studied by inhibitors, Western blot and siRNA. The results showed that the oxaliplatin resistance index of HCT116/Oxa cells was 10.2. HCT116/Oxa cells demonstrated stronger proliferation potential and anti-apoptotic capacity to oxaliplatin compared with HCT116 cells. Proteomic data demonstrated significant expression change of 717 genes in HCT116/Oxa cells, among which 399 genes were up-regulated while 318 ones down-regulated comparing with HCT116 cells. GO enrichment analysis showed that differentially expressed genes were mainly related to biological processes such as oxidative stress response, iron metabolism, lipid metabolism, apoptosis and cell cycle progression. Pathway analysis displayed notable changes of cell metabolism, ferroptosis, Nrf2-ARE signaling, fatty acid and glutathione metabolism in HCT116/Oxa cells. Quantitative results indicated that the expression of proteins directly related to ferroptosis, including glutathione peroxidase 4 (GPX4), glutamate-cysteine ligase regulatory subunit (GCLM), ferritin light chain (FTL), ferritin heavy chain (FTH1), heme oxygenase 1 (HMOX1), glutathione reductase (GSR) and NADH dehydrogenase 1 (NQO1) increased, while long chain fatty acid-CoA ligase (ACSL) 4 and ACSL1 decreased significantly in HCT116/Oxa cells. Functional studies showed that RSL3, a specific inhibitor of GPX4, decreased the viability of drug-resistant cells, improved lipid peroxidation, increased the concentration of ferrous ions, malondialdehyde, and decreased the concentration of glutathione (GSH). Western blot showed that the expressions of GPX4, FTH1, FTL and GSR increased in HCT116/Oxa, while ACSL4 decreased. RSL3 reversed the levels of GPX4, FTH1, FTL, GSR and ACSL4. It was further found that knockdown of GPX4 decreased the viability of drug-resistant cells, increased lipid peroxidation levels and decreased GSH concentration. These results suggest that ferroptosis resistance mediated by GSH/GPX4 pathway may be a potential mechanism of oxaliplatin resistance in HCT116/Oxa, and inhibition of GSH/GPX4 signaling could be an effective approach to reverse oxaliplatin resistance in colorectal cancer.

Polydatin (PD) is a natural active crystalline compound extracted from the roots and stems of Polygonum cuspidatum, and is a natural precursor of resveratrol. This study aims to investigate the therapeutic effects of PD on monosodium urate (MSU)-induced gouty arthritis in mice and its potential mechanisms. The animal experiment has been approved by the Ethics Committee of Nanjing University (approval number: 2407002). A gouty arthritis model was established by injecting 20 μL of MSU (25 mg·mL^-1) suspension into the mouse plantar. The effect of PD on pathological changes in the mouse plantar was evaluated. The treatment group received daily intraperitoneal injections of different doses of PD (low dose: 5 mg·kg^-1, medium dose: 10 mg·kg^-1, high dose: 20 mg·kg^-1) for 3 days before model induction. The thickness of the mouse plantar was measured and photographed at 3, 6, 9, 12, and 24 h after MSU suspension injection. Histopathological damage to the plantar tissue was observed using hematoxylin-eosin (H&E) staining. Immunohistochemistry and immunofluorescence were used to detect the expression of NLRP3 and CASP1 p20 to assess NLRP3 inflammasome activation in the plantar tissue. At the cellular level, lipopolysaccharide (LPS) combined with adenosine triphosphate (ATP)/MSU/nigericin was used to construct a cellular activation model of the NLRP3 inflammasome. ELISA was used to detect the effect of PD on interleukin-1β (IL-1β) secretion after NLRP3 inflammasome activation in macrophages. Flow cytometry was employed to measure CASP1 p20 activation in macrophages. Immunofluorescence was used to examine NLRP3 inflammasome assembly in macrophages. The results of the study indicate that, compared to the model group, the PD-treated group exhibited a significant reduction in the swelling of the mouse plantar. H&E staining showed a notable reduction in tissue damage in the mouse plantar, suggesting that PD has a therapeutic effect on plantar damage in mice. Immunohistochemistry and immunofluorescence results revealed a significant decrease in the expression of CASP1 p20 and NLRP3, indicating that PD significantly inhibits the activation of the NLRP3 inflammasome, thereby attenuating the local inflammatory response in the mouse plantar. At the cellular level, PD treatment significantly reduced the secretion of IL-1β and activation of CASP1 p20, both of which are mediated by NLRP3 inflammasome activation. Furthermore, NLRP3 inflammasome assembly was inhibited. In summary, PD exerts its anti-inflammatory effect by suppressing the assembly and activation of the NLRP3 inflammasome, reducing the production and release of the pro-inflammatory cytokine IL-1β, thereby alleviating joint damage in mouse gouty arthritis. This provides a novel strategy for the treatment of gout.

Four terpenoids were isolated from the neutral portion of petroleum ether extract of Artemisia annua by several chromatographic methods, such as silica gel, MCI Gel CHP-20, ODS, Sephadex LH-20 and semi-preparative HPLC. Their structures were identified by HR-MS and nuclear magnetic resonance spectroscopy. These compounds were defined as (1S,4S,5R,6S,9R,10R)-4-ethoxy-9,10-dimethyloctahydrofuro-(3,2-i)-isochromen-11(4H)-one (1), 3a,4,5,6,6a,7-hexahydro-3,6-dimethyl-9-methyl-2H-naphtho[8a,1-b]furan-2,8(3H)-dione (2), kobusone (3) and 1,2-campholide (4). Compound 1 is a new compound, of which the absolute configuration was established by single crystal X-ray crystallographic analysis. Compound 2 is a new natural product. Compounds 3 and 4 are first isolated from the Artemisia genus.

High-altitude sleep disturbance is a common acute high-altitude disease that can trigger physiological discomfort such as acute high-altitude reactions, with a lack of safe and effective preventive medications in clinical practice. Based on the gut-brain axis theory, this study designed and prepared a synbiotics combining Lactobacillus rhamnosus (LGG)-Lycium barbarum polysaccharide (LBP). First, the LGG-LBP synbiotics was prepared and evaluated. The mice were randomly divided into healthy, model, positive control (acetazolamide), LBP, LGG and LGG-LBP synbiotics group. After 7 days of administration, the mouse model of high-altitude sleep disturbance was established, the treating effects were evaluated through sleep duration, behavioral, hemogram test, and the content of tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS) in blood. 16S rRNA sequencing was used to analyze the changes of gut microbiota, and the pathological changes of small intestine were observed. The LGG-LBP synbiotics prolonged sleep duration, improved exploratory ability and short-term memory, promoted blood cell recovery. Moreover, LGG-LBP synbiotics enhanced the abundance of probiotics in the gut, and reduced intestinal inflammation. LGG-LBP synbiotics may be a potential prophylactic drug for high-altitude sleep disturbance. The animal operation was approved by the Ethics Committee of the Academy of Military Medical Sciences, Academy of Military Science (Approval number: IACUC-DWZX-2022-511). All experiments were conducted in accordance with relevant guidelines and regulations.

In the early stage of the project, it was found that natural pentacyclic triterpenes liquidambaric acid regulates the NEDD8 modification of Cullin2. This study aims to find more triterpenoid natural active molecules targeting Cullin family members and reveal its mechanism of action. Western blot was used to detect natural products that can significantly change the total protein NEDD8 modification and specific Cullin protein NEDD8 modification in cells; microscale thermophoresis (MST) was used to detect the direct binding of candidate small molecule oleanonic acid to TRAF family proteins, and the binding at the level of living cells was verified by cellular thermal shift assay (CETSA). Proximity ligation assay (PLA) was used to investigate the regulatory effect of oleanonic acid on the protein interaction between TNF receptor-associated factor 1 (TRAF1) and Cullin1 NEDD8 modified complex. Three pentacyclic triterpenoids were found to significantly inhibit NEDD8 modification in cells, among which oleanonic acid had the strongest effect on blocking NEDD8 modification. Different from the previous identification that liquidambaric acid regulates Cullin2/5, oleanonic acid can also specifically induce NEDD8-modified Cullin1 to transform into its unmodified form. And binding experiments showed that oleanonic acid could directly bind to TRAF1 at the level of cell lysate and living cells. Further mechanism studies found that oleanonic acid significantly changed the protein interaction between TRAF1 and Cullin1 NEDD8 modified complex. The above results indicate that oleanonic acid targets TRAF1 and regulates its interaction with NEDD8 modification complex to inhibit NEDD8 modification of Cullin.

In recent years, a large number of peptide compounds have been obtained from natural sources or synthesized chemically, which have attracted significant interest due to their high biological activity and low side effects. However, linear peptides encounter many challenges in the field of drug development because they are easily broken down by enzymes and do not pass through cell membranes well. Cyclic peptides, on the other hand, have a stable structure, strong binding to targets, and lower toxicity. They combine the advantages of natural peptides and small molecule drugs in terms of biological activity and drug metabolism, addressing the shortcomings of linear peptides and becoming increasingly important in drug research. This article focuses on the development history of cyclic peptides, discusses the sources, acquisition methods, and specific applications in the field of pharmacology in recent years, and prospects for their future development potential, aiming to provide a theoretical and practical basis for the clinical application of cyclic peptides.

Whole body irradiation (WBI) injury is defined multi-organ damages caused by whole-body exposure to ionizing radiation. The traditional radioprotective drug, amifostine, has significant adverse effects. Probiotics are reported to have radioprotective function, although their therapeutic efficacy is low due to poor gastrointestinal tolerance and the insufficient retention and colonization in the colon. In this study, chitosan/tannic acid double-layer-coated Lactobacillus reuteri was prepared, which was encapsulated in calcium alginate hydrogel microspheres to get an engineered probiotic-loaded microsphere formulation. The bilayer coating was confirmed by twice inversions of zeta potentials. Moreover, the coating improved bacterial adhesion and aggregation. Optical microscopy revealed the smooth morphology of microspheres, laser confocal imaging showed the uniform distribution of coated bacteria in microspheres, and scanning electron microscopy exhibited pores in the surface. The microspheres exhibited in vitro gastrointestinal resistance with rapidly swelling in the colonic environment to release bacteria. All the animal experiments were approved by Academy of Military Medicine Sciences (Approval No: IACUC-DWZX-2024-P510) and conducted in compliance with relevant guidelines. The 6.5 Gy whole-body irradiated mouse model was established. Starting from 2 days prior to irradiation, probiotic-loaded microspheres were administered via oral gavage consecutively for 6 days. Compared with the model group, the bacteria-loaded microspheres demonstrated protective effects on the hematopoietic system by promoting the recovery of red blood cells and platelets, maintaining the morphology of splenic red pulp and white pulp, and preserving bone marrow nucleated cells along with their proliferative capacity. Engineered probiotics have expanded the spectrum of radioprotective drugs, offering novel insights for the development of live biotherapeutic products aimed at preventing and treating radiation-induced injury.

Fusobacterium nucleatum (Fn) is closely associated with the occurrence and progression of colorectal cancer (CRC). The development of specific antibacterial agents targeting Fn is crucial for the prevention and treatment of CRC. Based on the preliminary phenotypic screening results from our research group, dimetridazole was successfully identified as a hit compound with antibacterial activity against Fn. In this preliminary structural optimization study, we designed and synthesized seven novel nitroimidazole derivatives comprising three structural types, followed by antimicrobial evaluation of all target compounds. Among them, compound CL6 exhibited excellent antibacterial activity against Fn (MIC = 0.5 μg·mL^-1) and demonstrated good selectivity towards intestinal bacteria and normal cells. Compound CL6 significantly inhibited the migration of CRC cells (HCT116) induced by Fn preliminary mechanistic studies suggest that compound CL6 disrupts the integrity of the Fn bacterial biofilm and cell wall, providing a promising lead compound for the development of novel anti-Fn drugs.

The methanol extract of Pteris wallichiana was separated and purified by MCI gel, sephadex LH 20, flash C18 and silica gel column chromatography combined with semi-pre HPLC. The chemical structures of the isolated compounds were identified by MS, IR, NMR, etc. Five sesquiterpene compounds were isolated from Pteris wallichiana and identified as 6,7-tetrahydrofuran-(2S, 3S)-pterosin C-3-O-β-D-(6′-acetyl)-Glu (1), (2S, 3S)-pterosin C-3-O-β-D-Glu (2), (2S)-pterosin A (3) and (2S)-13-hydroxyl-pterosin A (4), (2R, 3S)-2-hydroxyl-pterosin C (5). Compound 1 is a new sesquiterpene, compounds 3-5 were isolated for the first time. In vitro bioactivity assay showed that compound 1 was able to inhibit the proliferation of 4T1 and EMT6 cells, and possessed significant anti-triple-negative breast cancer bioactivity.

Tissue factor (TF), a transmembrane glycoprotein expressed in normal tissues, has a variety of physiological functions in embryonic development, hemostasis and non hemostasis pathways. Studies have found that TF is overexpressed in a variety of tumor tissues and promotes tumor progression. Kaplan Meier (K-M) survival analysis showed that high expression of TF gene was associated with poor prognosis in renal and pancreatic cancer. Therefore, TF has received extensive attention as a target of tumor immunotherapy, and a number of antibody-drug conjugates (ADC) drugs have entered the clinical research stage. In this paper, the gene structure, expression, biological function and the correlation with tumor of TF were systematically elaborated, and the direction of drug design for the new generation of TF-ADC was proposed, in order to provide theoretical support and development direction for the drug research and development of this target.