Radiation-induced intestinal injury (RIII) is a common syndrome in the radiotherapy of abdominal and pelvic malignant tumors, heavily influencing the living quality, but no specific clinical regimens are available. Inulin is a naturally soluble dietary fiber. Clostridium butyricum (Cb) is a strict Gram-positive anaerobic bacillus, which can secrete and produce a large number of butyric acid, improving the intestinal barrier function and reducing the growth and colonization of opportunistic bacteria. A synbiotic of inulin gels (IG) and Cb exerts the synergistic effect of prebiotic intestinal retention and probiotics. In this study, an oral synbiotic of IG and Cb was applied for the prevention and treatment of RIII and the mechanisms were explored. Animal experiments were approved by the Ethics Committee of the Academy of Military Medical Sciences and the experiments were conducted in accordance with relevant guidelines and regulations (authorizing number: IACUC-DWZX-2022-525). An RIII mouse model was established after whole abdominal γ-ray radiation of 13 Gy. The synbiotic was intragastrically administered to the mice 1 h pre-radiation. Compared to the models, the mice of the synbiotic group had more regenerated intestinal crypts, longer villus and colon, and more tight junction protein on day 3.5. Moreover, the mice of the synbiotic group had an obvious increase in the relative abundance of gut microbiota on day 7, especially the amounts of multiple probiotics and short-chain fatty acid metabolites. On day 14, the mice of the synbiotic group had highly low permeability of intestinal mucosa according to the fluorescence labeling experiment, which was close to the normal level. Moreover, the mice of the synbiotic group showed a high decrease of proinflammatory factors including tumor necrosis factor-α and interleukin-6, close to the normal levels. Therefore, the oral synbiotic can alleviate the syndromes of RIII and improve the repair of damaged intestinal and colon tissues, and its protective effect is better than IG and Cb. The oral synbiotic is a safe and effective biological drug against RIII.

Attenuated Salmonella typhimurium VNP20009 is a novel oncolytic bacterium with high tumor-targeting properties. One of its anti-tumor mechanisms is the induction of tumor cell apoptosis, although the specific molecular mechanisms remain unclear. Melanoma, the deadliest form of skin cancer, is associated with significant challenges, such as severe side effects and high recurrence rates in current treatments. This study used the B16F10 mouse melanoma cell line as a model to explore the regulatory mechanism of VNP20009-induced apoptosis in melanoma cells. The results showed that VNP20009 significantly induced apoptosis in B16F10 cells in a time- and concentration-dependent manner. Transcriptomic analysis revealed that the p53 signaling pathway was significantly enriched in the VNP20009-treated group, suggesting that this pathway might mediate the pro-apoptotic effects of VNP20009. Further investigations demonstrated that VNP20009 induces apoptosis by activating key genes in the p53 pathway, including PUMA, and its upstream and downstream molecules, such as p53, CytC, CASP9, and CASP3, forming a cascade reaction. In conclusion, this study elucidates the molecular mechanism by which VNP20009 induces apoptosis in B16F10 melanoma cells through the p53-PUMA axis, providing new theoretical insights for melanoma treatment based on attenuated Salmonella bacteria.

Tigliane-type diterpenoids are the main active components of croton and a diverse group of macrocyclic diterpenes with diverse biological activities. Their anti-HIV activity have unique advantages and great potential in the treatment of acquired immunodeficiency syndrome (AIDS), and were expected to be developed as a candidate drug for clinical trials of anti-AIDS. This article reviews the research progress on the components of tigliane-type diterpenoids in croton and their anti-HIV activity in recent years, conducts systematic comparisons, summarizes the currently discovered tigliane-type diterpenoids in croton. Their chemical structures and biological activity relationship were summarized. The research progress on the mechanism of tigliane-type diterpenoids' anti-HIV activity was briefly discussed. This research could provide guidance for further development and utilization of coumarin type diterpenes, as well as new ideas for the application of croton.

With the increasing abuse of antibiotics and the growing resistance of bacteria, it is urgent to find new antibacterial agents. Numerous constituents of traditional Chinese medicine exhibit pronounced antibacterial, anti-inflammatory, and antioxidant pharmacological properties, often operating through multiple mechanisms, thereby positioning them as a vital source for the development of novel antibacterial agents in the future. Nevertheless, the antibacterial constituents of traditional Chinese medicine exhibit challenges such as inadequate stability, low solubility, and suboptimal intelligent release capabilities, which hinder their extensive application in antibacterial formulations. Metal-organic framework materials serve as highly effective drug carriers for antibacterial constituents of traditional Chinese medicine, attributed to their high specific surface area, elevated porosity, controllable pore dimensions, and responsive release properties. Furthermore, they not only enhance the stability and solubility of these antibacterial constituents while also exhibiting inherent antibacterial activity and responsive release capabilities. This paper presents a comprehensive overview of bacterial resistance mechanisms and the action pathways of antibacterial constituents of traditional Chinese medicine against resistant bacteria. Additionally, it highlights the current advancements in metal-organic framework materials and their application in the delivery systems for these antibacterial constituents, aiming to provide valuable insights for the research and innovation of formulations based on traditional Chinese medicine.

Combined radiation and burn injury (CRBI) is induced by simultaneous or sequential ionizing radiation damage and skin burns. CRBI weakens the immune ability, leading to drug-resistant bacterial infections and delayed wound healing. Bdellovibrio-and-like organisms (BALO) are naturally predatory bacterium that can prey on most Gram-negative bacteria by entering the periplasmic space of their prey and degrading the biomolecules of host cells. In this study, we combined gelatin, calcium alginate, and activated BALO water samples to form bio-inks to three-dimensional (3D)-print BALO-loaded hydrogels (TDBG) for the treatment of CRBI combined with multidrug-resistant Acinetobacter baumannii (MRAB) infection. The freeze-dried 3D-printed hydrogel exhibited a 3D network structure attached with gelatin films, and owned good printability and biocompatibility. The printability improved adaptation to wound shapes for the personalized treatment of infected wounds. The 3D network structure allowed the surviving and motion of BALO, favoring its high predatory activity. All animal experiments were approved by the Ethics Committee of Academy of Military Medical Sciences, and the experiments were conducted in accordance with relevant guidelines and regulations (approval number: IACUC-DWZX-2022-834). TDBG treatment improved wound healing by accelerating the mouse wound closure rate of CRBI combined with MRAB infection, reducing the expression of pro-inflammatory cytokines in the wound tissues, and increasing collagen deposition. This study expands the application scope of live biological products and provides a basis for their development and clinical applications.

Neurodegenerative diseases are one of the leading causes of death and disability worldwide, with complex pathogenesis and lacking effective therapeutic drugs. Increasing researches have shown that most neurodegenerative diseases involve abnormalities in iron homeostasis and activation of immune cells in the brain. Iron accumulation in microglia promotes ferroptosis, leading to cellular dysfunction and death. In contrast, inhibiting ferroptosis can alleviate neuroinflammation, protect neurons, and slow disease progression, highlighting the pivotal roles of ferroptosis and neuroinflammation in neurodegenerative diseases. This review summarizes the roles of ferroptosis and neuroinflammation in neurodegenerative diseases, further discusses the related targets regulating these processes, and reviews the therapeutic potential of drugs targeting ferroptosis and neuroinflammation in neurodegenerative diseases. This review aims to provide novel targets and therapeutic drugs for the treatment of neurodegenerative disease, offering new strategies for clinical management and improving the symptoms and prognosis of neurodegenerative disease.

2',4'-Dimethoxychalcone (DMC) is a structural modifier of carvacrol B. In this study, gastric cancer cells MGC-803 and HGC-27 were used as the subjects to investigate the anti-tumor effect and mechanism of DMC on gastric cancer (GC) cells both in vitro and in vivo. DMC inhibited cell viability and cell proliferation and promoted cell apoptosis in GC cells, detected by CCK-8 assay, EdU staining and Annexin V-FITC/PI double-staining flow cytometry. A nude mouse model of GC cell xenograft was constructed by subcutaneous injection with MGC-803 cells, for measuring the effect of DMC on the growth of GC in vivo, and DMC inhibited the growth of subcutaneous transplantation tumor in nude mice. The animal experiments were approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine under the ethical number PZSHUTCM2310110002. The effect of DMC on the RNA expression of MGC-803 cells was detected by RNA-seq assay, and it was found that the biological function of DMC was enriched in glycolysis. DMC inhibited the glucose uptake capacity and lactate production and efflux of gastric cancer cells, detected by using 2-NBDG probe with flow cytometry and lactate (LD) test kit. Western blot assay was performed to detect the protein expression of proliferation, apoptosis, and glycolysis-related proteins in gastric cancer cells, and the results demonstrated that DMC up-regulated the protein expression of cleaved caspase-9, cleaved caspase-3, cleaved PARP, down-regulated Ki-67 protein expression, and inhibited the protein expression of c-Myc, LDHA, GLUT3, PDHK1 and MCT1 in gastric cancer cells. The Seahorse energy metabolism analyser was used to measure the rate of glycolysis, and it was found that DMC could down-regulate the basal glycolysis rate and compensatory glycolysis in gastric cancer cells. The c-Myc overexpressing cell line MGC-803 was used in the reversal experiment to further confirm that DMC suppressed gastric cancer growth through inhibiting c-Myc mediated glucose uptake and glycolysis. In conclusion, DMC may inhibit the protein expression of c-Myc and its target glycolysis-related genes, suppressed c-Myc-mediated glucose uptake and glycolysis in gastric cancer cells, thereby inhibited the cell proliferation and promoted cell apoptosis of gastric cancer cells, and thus finally inhibited the growth of gastric cancer in vivo and in vitro.

Liver fibrosis is a chronic liver injury caused by various pathogenic factors, leading to excessive accumulation of extracellular matrix such as collagen. It represents a common pathological hallmark during the progression of most chronic liver diseases. However, there is currently no universally recognized specific and effective drug for the clinical treatment of liver fibrosis. Therefore, this study investigates the effects of Alisma Rhizoma on bile duct ligation (BDL)-induced liver fibrosis and explores the potential pharmacological mechanisms. The animal experimental protocol was reviewed and approved by the Animal Welfare and Ethics Committee of Shanghai University of Traditional Chinese Medicine (registration No. PZSHUTCM2303280007), in compliance with relevant animal welfare and ethical standards. Mice were subjected to BDL to induce liver fibrosis. Mice were divided into five groups: sham operation group (Sham), model group (BDL), ethanol extract protection group (BDL+EE, 1.6 g·kg^-1), water extract protection group (BDL+WE, 4.0 g·kg^-1), obeticholic acid protection group (BDL+OCA, 10 mg·kg^-1). The results showed that both of EE and WE could attenuate BDL-induced liver fibrosis as evident by reduced serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT) activities, total bile acids (TBA) levels, and improved pathological conditions such as cholestasis, collagen deposition, inflammatory cell infiltration, and liver tissue necrosis. Notably, EE showed better efficacy than WE. Further studies showed that EE improved liver fibrosis dose dependently. EE treatment impaired the bile acids homeostasis in serum and liver, and recovered the hepatic mRNA expression of farnesoid X receptor (FXR) as well as the downstream genes including small heterodimer partner (SHP), cholesterol 7-alpha hydroxylase (CYP7A1) and bile salt export pump (BSEP). Further study also proved that the four major triterpenes in EE increased the transcriptional activities of FXR in vitro. This study provides a theoretical basis for the clinical application of Alisma Rhizoma in the prevention and treatment of liver fibrosis.

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.

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.