Pig hoof nails represent an underutilized byproduct of pork processing. According to traditional Chinese medical theory, pig hoof nails are believed to contain compounds that may exhibit pharmacological properties such as promoting hemostasis, aiding coagulation, and reducing inflammation, which could potentially benefit the healing of skin wounds. In this study, different molecular masses of hoof nail peptides (HNPs) were extracted from porcine hoof nails. Wound healing assays were employed to identify HNPs that significantly promote fibroblast proliferation and collagen synthesis across three molecular weight ranges: less than 10 kDa (HNP Ⅰ), 3-10 kDa (HNP Ⅱ), and more than 10 kDa (HNP Ⅲ). We formulated HNP Ⅱ into a temperature-sensitive gel designed to conform to uneven wound surfaces. This supports the development and application of hoof nail peptide formulations. The animal operation was approved by the Laboratory Animal Ethics Committee of Southwest University (No: IACUC-20231218-01). All experiments were conducted in accordance with relevant guidelines and regulations.

Simvastatin is a class of hydroxymethylglutaryl coenzyme A reductase inhibitors used to lower blood lipids and prevent atherosclerosis, but its poor water solubility and low bioavailability seriously affect clinical efficacy. In this experiment, simvastatin solid dispersions were prepared using supercritical CO₂ antisolvent method, and the prescription process parameters were optimized by Box-Benhnken response surface methodology, to improve the dissolution characteristics. The Box-Benhnken response surface methodology optimization experimental scheme took in vitro cumulative dissolution degree of simvastatin (dependent variable) as the evaluation index, investigated the influence of four factors (independent variables) including pressure, temperature, carrier type, and CO₂ outflow rate on the preparation of simvastatin solid dispersions by supercritical CO₂ antisolvent technology. Next, the particle size (PZ), infrared (IR) absorption spectrum, differential scanning calorimetry (DSC), X-ray diffraction (XRD), and scanning electron microscopy (SEM) characterization were performed on the preferred process conditions for simvastatin dispersion. Based on these findings, further research was conducted on the in vitro dissolution properties of the product as well as the relative bioavailability after oral administration to rats. Animal experiments were approved by the Animal Ethics Committee of Jiangsu College of Food and Drug Engineering (No. JSSPYPCAC2024). The results showed that the optimal process parameters for preparing simvastatin solid dispersions by critical CO₂ antisolvent technology were pressure 12.00 Mpa, temperature 35.0 ℃, CO₂ flow rate 3.3 L·min^-1, F-127/PVP K23-27 (1∶10, w/w). The prepared solid dispersion was in a loose powder form, which appears as overlapping flaky aggregates under SEM. The laser particle size analysis showed a D50 value of 0.78 μm. DSC and XRD indicated that simvastatin existed in an amorphous form within the dispersion. The in vitro cumulative dissolution value after 60 minutes was 89.87%, and the relative bioavailability of oral administration to rats was 226.6%. In summary, this study successfully applied supercritical CO₂ antisolvent method to prepare simvastatin solid dispersions, simplified the experimental steps and optimized process routes using Box-Benhnken response surface methodology. The prepared simvastatin solid dispersions exhibited fast dissolution rates and high cumulative dissolution degrees, which can be used as a prescription strategy for developing novel oral simvastatin formulations with high bioavailability.

Chinese medicine supramolecular is an important material form and effective form of Chinese medicine compound. The form and activity of supramolecular are affected by many factors, and the mixing order is one of the key factors. In this study, dynamic light scattering (DLS), scanning electron microscopy (SEM), turbidimeter and Tyndall effect were used to observe the differences of micromorphology and physical properties of three different groups of Coptidis Rhizoma group + Gardeniae Fructus + Scutellariae Radix (CGS), Coptidis Rhizoma group + Scutellariae Radix + Gardeniae Fructus (CSG) and Scutellariae Radix + Gardeniae Fructus + Coptidis Rhizoma group (SGC). The main components of the three groups of decoction were analyzed by high performance liquid chromatography (HPLC). The antibacterial activity of the three groups of decoction was preliminarily compared by in vitro Staphylococcus model, and the dynamic formation process of supramolecules in the three groups of decoction was preliminarily discussed by molecular dynamics simulation (MDS) technology. The results showed that different mixing sequences led to changes in the morphology of supramolecular parts of Huanglian Jiedu Decoction, with significant differences in macroscopic phenomena, electron microscopy, particle size, potential and turbidity, and significantly affected the original antibacterial activity. In addition, the results of molecular dynamics simulation also show that the different mixing order of the main components of Huanglian Jiedu Decoction will significantly affect its self-assembly behavior. This study is to further explore the factors affecting the formation of supramolecular substance basis of Huanglian Jiedu Decoction, also provides a reference for the clinical rational administration of Huanglian Jiedu Decoction formula granules.

Live biotherapeutic products (LBPs) represent a distinct category of biological products containing viable organisms, such as bacteria, utilized for the prevention and treatment of human diseases (excluding vaccines). Presently, research and development efforts in LBPs are predominantly centered on live bacteria. Compared to traditional drugs, the LBPs demonstrate unique characteristics, including replicability, target specificity, and responsiveness. Owing to these properties, LBPs have emerged as hotspots in the development of specialized treatments for various major diseases, with applications spanning malignant tumors, metabolic disorders, inflammatory bowel diseases, genetic defects, and more. Nevertheless, natural bacteria face inherent limitations—such as low activity, instability, and safety concerns—that hinder their pharmacological potential. As a result, engineering strategies have become essential for enhancing the properties of bacteria and facilitating their clinical applications. This article delves into recent advancements in LBPs derived from engineered bacteria, offering a systematic review of reported engineering strategies, which are broadly categorized into chemical, physical, and genetic modifications. The findings indicate that no single engineering approach can comprehensively address all the challenges associated with converting viable bacteria into effective LBPs. To overcome this limitation, a concept of "multi-engineered bacteria" is introduced. This framework advocates for the integration of physical, chemical, and biological engineering strategies to develop next-generation LBPs with enhanced functionality and clinical potential. This article provides a concise review of current research on LBPs based on engineered bacteria and outlines forward-looking perspectives for advancing their development through innovative engineering approaches.

Bacterial infectious diseases persistently pose severe threats to human health, development of livestock and aquaculture industries, and ecological stability. The extensive use of conventional antibiotics has led to increasingly critical issues of bacterial resistance, making the development of novel and effective strategies for preventing and treating bacterial infections an urgent priority. Bdellovibrio bacteriovorus, as a genus of parasitic bacteria that prey on other bacteria, exhibits lytic activity against various pathogenic species and demonstrates potential for combating bacterial infections. However, the direct application of B. bacteriovorus suspensions or powders faces challenges including rapid clearance, susceptibility to immune system elimination, difficulty in maintaining their vitality, and poor user compliance. Recent advancements in engineered B. bacteriovorus technology have created new opportunities for more precise and efficient utilization of these predators in infection control. This paper reviews recent advances in engineered B. bacteriovorus for bacterial infection control, with particular emphasis on engineering strategies based on formulation design, surface modification, and genetic editing, along with their therapeutic applications. The review aims to provide valuable insights for advancing research on engineered B. bacteriovorus technologies.

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.

Oral probiotics are susceptible to the gastrointestinal environment, so the number of probiotics reaching the intestine is small and difficult to colonize, limiting the application of probiotic therapy. In this study, Lactobacillus rhamnosus (LGG), a common probiotic, was chosen as a model, and layer-by-layer encapsulated LGG-loaded porous microspheres with glycol chitosan (GCS) and sodium alginate (SA) were prepared to investigate it's in vitro properties. Poly-L-lactic acid porous microspheres (PLPM) were prepared by the complex milk-solvent evaporation method, with rounded morphology, uniform size, open and connected porous structure, and the average particle size of 138.5 μm. The PLPM were co-incubated with LGG for 8 h at 37 ℃ to obtain the LGG-loaded porous microspheres (LPM) with high bacterial loadings. The surface of the LPM were wrapped with GCS and SA layer by layer by electrostatic action to obtain the layer-by-layer encapsulated LGG-loaded porous microspheres with GCS and SA (AGLPM). In vitro experiments demonstrated that AGLPM could tolerate simulated gastric fluid at pH 1.2 and simulated intestinal fluid at pH 7.4 for 2 h, and its stability was significantly better than that of bare LGG. AGLPM was a better probiotic dosage form.

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.

Radiation enteritis (RE) is the most common complication during radiotherapy, although safe and effective drugs for the treatment of RE are deficient. Probiotics have been demonstrated to own antiradiation function. Synbiotics are composed of probiotics and prebiotics, which enhance the ability of probiotics. Here, inulin gel (IG) that own the ability to resist gastric acid and retain in the colon were used to load three types of probiotics, including Bacillus cereus (BC), Bacillus licheniformis (BL), and Lactobacillus reuteri (LR), respectively. The RE pharmacodynamic studies of them were conducted. The probiotics were embedded in the IG by scanning electron microscopy and confocal laser scanning microscopy. IG promoted the growth of probiotics compared to probiotics alone. 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 (No. IACUC-DWZX-2024-P510). Mice experienced the whole abdominal irradiation of 13 Gy γ rays to form RE models. Compared to correspondingly probiotics alone, all probiotic-loaded IG synbiotics had the better prevention and treatment efficiencies in repairing the intestinal barriers, improving the structure of crypts, oxidative stress, inflammation, and imbalanced gut microbiota and promoting the recovery of intestinal villus, where the Bacillus cereus-loaded IG was the best. This study provides a novel therapeutic for the prevention and treatment of RE.

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.