In this study, RAW264.7 cells were employed to investigate the effects of honey-processed Astragalus on their energy metabolism and polarization, and explore the scientific connotation of the enhanced efficacy of honey-processed Astragalus on invigorating spleen-stomach and replenishing Qi. The medicated sera were prepared by intragastric administration of rats with water extracts of crude and honey-processed Astragalus, and the composition changes of medicated sera of crude and honey-processed Astragalus were analyzed by using LC-MS technology. The cell survival rates were detected and the concentrations of medicated sera were screened through CCK-8 assay. The differences of cell phagocytic rates, ATP energy metabolism, and NO secretion between crude and honey-processed Astragalus were evaluated by using neutral red phagocytosis assay, ATP detection kit, and NO detection kit. The effects of crude and honey-processed Astragalus on TNF-α secretion of RAW264.7 cells were detected by employing ELISA kit. The effects of crude and honey-processed Astragalus on polarization of RAW264.7 cells were evaluated by utilizing flow cytometry. The differential metabolites related to glycolysis in cell lysates and culture media were screened by using LC-MS technology. The experiment was approved by the experimental animal ethics committee from Shanxi University of Chinese Medicine (No. AWE202407352). The results showed that the contents of betaine, amino acids, and ononin in the prepared medicated sera of rats treated by intragastric administration with water extracts of honey-processed Astragalus increased compared to those in crude one. The results of CCK-8 experiment showed that there were no cytotoxic effects on RAW264.7 cells in the medicated sera of crude and honey-processd Astragalus at different concentration. The phagocytic index and ATP yield both increased to varying degrees after administration of the medicated sera to cells. The secretion of NO in normal and inflammatory cells increased and decreased respectively, and the effect of honey-processed Astragalus was better than that of crude one. The results of ELISA kit showed that the medicated sera of both crude and honey-processed Astragalus could promote the secretion of TNF-α in a concentration-dependent manner, and the promoting effect of honey-processed Astragalus was stronger than that of crude one. The results of flow cytometry showed that the medicated sera of both crude and honey-processed Astragalus could promote M1-type polarization and inhibit M2-type polarization of RAW264.7 cells, and the effect of honey-processed Astragalus was better than that of crude one. Compared to crude Astragalus, the metabolites related to glycolysis in the cell lysates and culture media of the medicated sera of honey-processed Astragalus were generally on the rise, indicating that the effect on promoting glycolysis of honey-processed Astragalus was better than that of crude one. In summary, honey-processed Astragalus can promote the polarization and energy metabolism of RAW264.7 cells, and participate in positive immune regulation, which is correlated with its enhanced effect of invigorating spleen-stomach and replenishing Qi.

OBJECTIVE: To evaluate precise assessment methods for predicting the optimal acetabular cup size in total hip arthroplasty (THA). METHODS: A clinical data of 73 patients (80 hips) who underwent primary THA between December 2022 and July 2024 and met the inclusion criteria was analyzed. There were 39 males and 34 females with an average age of 66.3 years (range, 56-78 years). Among them, 66 cases were unilateral THA and 7 were bilateral THAs. There were 29 patients (34 hips) of osteoarthritis, 35 patients (35 hips) of femoral neck fractures, and 9 patients (11 hips) of osteonecrosis of the femoral head. Based on anteroposterior pelvic X-ray films, three methods were employed to predict acetabular cup size, including preoperative template planning, radiographic femoral head diameter (FHD) measurement, and intraoperative FHD measurement. The predicted acetabular cup sizes from these methods were compared with the actual implanted sizes. RESULTS: The predicted acetabular cup sizes using the preoperative template planning, radiographic FHD measurement, and intraoperative FHD measurement were (51.25±2.81), (49.72±3.11), and (49.90±2.74) mm, respectively, compared to the actual implanted cup size of (50.57±2.74) mm, with no significant difference ( P>0.05). Regarding agreement with the actual implanted cup size, the preoperative template planning achieved exact matches in 35 hips (43.75%), one-size deviation in 41 hips (51.25%), and two-size deviations in 4 hips (5%); the radiographic FHD measurement achieved exact matches in 12 hips (15%), one-size deviation in 57 hips (71.25%), and two-size deviations in 11 hips (13.75%); and the intraoperative FHD measurement achieved exact matches in 26 hips (32.5%), one-size deviation in 52 hips (65%), and two-size deviations in 2 hips (2.5%). There were significant differences in agreement distributions between the three methods and the actual implanted cup sizes ( H=18.579, P<0.001). CONCLUSION The intraoperative FHD measurement, as a simple, cost-effective, and accurate method, effectively guides acetabular cup selection, reduces the risk of prosthesis wear, enhances postoperative joint stability.
Humans ; Arthroplasty, Replacement, Hip/instrumentation* ; Male ; Female ; Middle Aged ; Acetabulum/diagnostic imaging* ; Aged ; Hip Prosthesis ; Prosthesis Design ; Femur Head/surgery* ; Osteoarthritis, Hip/surgery* ; Radiography ; Femoral Neck Fractures/surgery* ; Femur Head Necrosis/surgery*

Neddylation is a crucial posttranslational modification that involves the attachment of neural precursor cell-expressed developmentally downregulated protein 8 (NEDD8) to a lysine residue in the substrate via the sequential actions of the E1 NEDD8-activating enzyme (NAE) (E1), E2 NEDD8-conjugating enzyme (E2), and E3 NEDD8-ligase (E3). The most extensively studied substrates of neddylation are members of the cullin family, which act as scaffold components for cullin ring E3 ubiquitin ligases (CRLs). Since cullin neddylation activates CRLs, which are frequently overactive in tumors, inhibiting neddylation has emerged as a promising strategy for developing novel antitumor therapies. This review explores the antitumor effects of inhibiting neddylation that leads to the inactivation of CRLs and provides a summary of known inhibitors that target protein-protein interactions (PPIs) within the neddylation enzymatic cascade.

In this study, we employed a combination of genome mining and heteronuclear single quantum coherence (HSQC)-based small molecule accurate recognition technology (SMART) technology to search for fernane-type triterpenoids. Initially, potential endophytic fungi were identified through genome mining. Subsequently, fine fractions containing various fernane-type triterpenoids were selected using HSQC data collection and SMART prediction. These triterpenoids were then obtained through targeted isolation and identification. Finally, their antifungal activity was evaluated. As a result, three fernane-type triterpenoids, including two novel compounds, along with two new sesquiterpenes and four known compounds were isolated from one potential strain, Diaporthe discoidispora. Their structures were elucidated through analysis of high-resolution electrospray ionization mass spectrometry (HR-ESI-MS) and nuclear magnetic resonance (NMR) spectroscopic data. The absolute configurations were determined using single-crystal X-ray diffraction analysis and electron capture detector (ECD) analysis. Compound 3 exhibited moderate antifungal activity against Candida albicans CMCC 98001 and Aspergillus niger.
Triterpenes/isolation & purification* ; Antifungal Agents/isolation & purification* ; Molecular Structure ; Candida albicans/drug effects* ; Ascomycota/genetics* ; Magnetic Resonance Spectroscopy ; Aspergillus niger/drug effects* ; Genome, Fungal ; Microbial Sensitivity Tests

OBJECTIVE: The relationship between non-high-density lipoprotein (NHDL) cholesterol to high-density lipoprotein cholesterol (HDL-C) ratio (NHHR) and stoke remains unknown. This study aimed to evaluate the association between the adult NHHR and stroke occurrence in the United States of America (USA). METHODS: To clarify the relationship between the NHHR and stroke risk, this study used a multivariable logistic regression model and a restricted cubic spline (RCS) model to investigate the association between the NHHR and stroke, and data from the National Health and Nutrition Examination Survey (NHANES) from 2005 to 2018. Subgroup and sensitivity analyses were conducted to test the robustness of the results. RESULTS: This study included 29,928 adult participants, of which 1,165 participants had a history of stroke. Logistic regression analysis of variables demonstrated a positive association between NHHR and stroke ( OR 1.24, 95% CI: 1.03-1.50, P = 0.026). Compared with the lowest reference group of NHHR, participants in the second, third, and fourth quartile had a significantly increased risk of stroke after full adjustments ( OR: 1.35, 95% CI: 1.08-1.69) ( OR: 1.83, 95% CI: 1.42-2.36) ( OR: 2.04, 95% CI: 1.50-2.79). In the total population, a nonlinear dose-response relationship was observed between the NHHR and stroke risk ( P non-linearity = 0.002). This association remained significant in several subgroup analyses. Further investigation of the NHHR may enhance our understanding of stroke prevention and treatment. CONCLUSION Our findings suggest a positive correlation between the NHHR and an increased prevalence of stroke, potentially serving as a novel predictive factor for stroke. Timely intervention and management of the NHHR may effectively mitigate stroke occurrence. Prospective studies are required to validate this association and further explore the underlying biological mechanisms.
Humans ; Stroke/blood* ; United States/epidemiology* ; Male ; Female ; Middle Aged ; Cross-Sectional Studies ; Nutrition Surveys ; Adult ; Aged ; Cholesterol, HDL/blood* ; Cholesterol/blood* ; Risk Factors

OBJECTIVE: To reveal the effects and potential mechanisms by which synaptic vesicle glycoprotein 2A (SV2A) influences the distribution of amyloid precursor protein (APP) in the trans-Golgi network (TGN), endolysosomal system, and cell membranes and to reveal the effects of SV2A on APP amyloid degradation. METHODS: Colocalization analysis of APP with specific tagged proteins in the TGN, ensolysosomal system, and cell membrane was performed to explore the effects of SV2A on the intracellular transport of APP. APP, β-site amyloid precursor protein cleaving enzyme 1 (BACE1) expressions, and APP cleavage products levels were investigated to observe the effects of SV2A on APP amyloidogenic processing. RESULTS: APP localization was reduced in the TGN, early endosomes, late endosomes, and lysosomes, whereas it was increased in the recycling endosomes and cell membrane of SV2A-overexpressed neurons. Moreover, Arl5b (ADP-ribosylation factor 5b), a protein responsible for transporting APP from the TGN to early endosomes, was upregulated by SV2A. SV2A overexpression also decreased APP transport from the cell membrane to early endosomes by downregulating APP endocytosis. In addition, products of APP amyloid degradation, including sAPPβ, Aβ _1-42, and Aβ _1-40, were decreased in SV2A-overexpressed cells. CONCLUSION These results demonstrated that SV2A promotes APP transport from the TGN to early endosomes by upregulating Arl5b and promoting APP transport from early endosomes to recycling endosomes-cell membrane pathway, which slows APP amyloid degradation.
Amyloid beta-Protein Precursor/genetics* ; Membrane Glycoproteins/genetics* ; Animals ; Protein Transport ; Nerve Tissue Proteins/genetics* ; Humans ; Mice ; Endosomes/metabolism* ; trans-Golgi Network/metabolism*

Objectiveβ‑Site APP cleaving enzyme 1 (BACE1) is a rate-limiting enzyme involved in the formation of amyloid plaques in Alzheimer’s disease (AD), and its expression and activity play a crucial role in the development of AD. The interacting protein of BACE1 can directly or indirectly regulate BACE1 in the transcription, translation, modification, intracellular transport and other links of BACE1 by directly binding, indirectly binding, and participating in various cell signal transduction pathways, so as to participate in the occurrence of AD and the process of disease. This study aimed to screen and validate the interacting proteins of BACE1, providing new insights into the mechanisms of amyloid plaque formation. MethodsCo-immunoprecipitation (Co-IP) and mass spectrometry (MS) were used to enrich and identify BACE1 interacting proteins in the hippocampus of wild type (WT) mice and AD model mice. For candidate BACE1 interacting proteins, GO enrichment analysis and KEGG pathway enrichment analysis were used to explore the subcellular localization, molecular function, participating biological processes and participating signaling pathways of BACE1 interacting proteins. The protein-protein interaction (PPI) network of BACE1 was further constructed to explore the potential proteins interacting with BACE1. By searching the mouse genomeinformation (MGI) website and NCBI database, the more reliable proteins among the potential BACE1 interacting proteins were screened. Co-IP assay and immunofluorescence confocal technology were used to preliminarily verify the interaction between the proteins, and the changes in protein expression levels of the interacting proteins in AD cell models were explored. ResultsA total of 614 differentially expressed proteins interacting with BACE1 were identified in AD group. GO enrichment analysis showed that the BACE1 interacting proteins in the AD group were mainly located in membrane organelles such as Golgi apparatus, endoplasmic reticulum, endosome, lysosome and vesicles, which had molecular functions such as ion channel regulation, protein kinase activity, transcription factor binding and passive transmembrane transporter activity. It is mainly involved in the biological processes of immune response regulation cell surface receptor signaling pathway, targeting Golgi vesicles transport, circadian rhythm regulation, Purkinje cell layer development, etc. KEGG analysis showed that BACE1 interacting proteins in AD were mainly involved in the PI3K-Akt signaling pathway, mTOR signaling pathway and other neurodegenerative disease-related pathways. The PPI network of BACE1 showed that a total of 12 proteins were identified as high confidence binding proteins, including PRNP, APOE, SYP, NSF, NUMB, SNAP91, HSP90aa1, UCHL1, BIN1, SNX27, Rheb, Ap2m1, of which, NSF, NUMB, SNAP91, HSP90aa1 were newly identified candidate proteins. After further verification, we found that NSF not only interacts with BACE1, but also interacts with amyloid precursor protein (APP), the substrate of BACE1, and the expression level of NSF is up-regulated in the AD cell model constructed by Aβ₄₂ induction. ConclusionBACE1 binding proteins participate in multiple AD-associated biological processes and signal pathways. NSF is a newly identified BACE1 binding protein that interacts with BACE1, and the protein expression level of NSF is up-regulated in the AD cell model. It is predicted that the interaction between NSF and BACE1 is involved in regulating the course of AD, providing a new target and direction for the study of the mechanism of AD.

OBJECTIVE To explore in vitro dissolution and in vivo pharmacokinetics of Luteolin solid dispersion in Beagle dogs. METHODS The dissolution of Luteolin solid dispersion was investigated according to the second method （paddle method） of the “dissolution determination method” in the 2020 edition of Chinese Pharmacopoeia （Part Ⅳ）. UPLC-MS/MS method was established to determine the concentration of luteolin in the plasma of Beagle dogs. Twelve Beagle dogs were randomly divided into luteolin group and Luteolin solid dispersion group， with 6 dogs in each group. They were given relevant medicine orally at the dose of 10 mg/kg luteolin. Blood was collected before medication （0 h）， at 5， 10， 15， 30， 45 min and 1， 2， 4， 6， 8， 10， 12， 24， 48 h after administration. After protein precipitation with acetonitrile， the blood concentration of luteolin in Beagle dogs was determined by UPLC-MS/MS and the major pharmacokinetic parameters were calculated with non-compartmental models by using DAS 3.2.8 pharmacokinetic software. RESULTS The dissolutions of Luteolin solid dispersion in purified water and 0.1% sodium dodecyl sulfate solution was significantly higher than those of luteolin； the dissolution rate reached 95% in 0.1% sodium dodecyl sulfate solution for 120 minutes. The peak concentration （cmax） of luteolin in the Luteolin solid dispersion group of Beagle dogs was 5.62 times higher than the luteolin group， and the relative bioavailability was 348%. Compared with luteolin group， cmax and the area under the drug time curve of luteolin in the Luteolin solid dispersion group of Beagle dogs were significantly increased， while the apparent distribution volume was significantly reduced （P＜0.05）. CONCLUSIONS Luteolin solid dispersion can improve in vitro dissolution and bioavailability of luteolin in Beagle dogs.

Radiotherapy is an important means to treat lung cancer,but it is easy to cause lung injury and reduce the quality of life of patients.Flash radiotherapy(FLASH-RT)has attracted attention due to its extremely short radiation duration and high dose rate,which can reduce toxicity of normal tissue while ensures treatment intensity of tumor.Whether Flash-RT can reduce radiation-induced lung injury has become an important research topic in recent years.Based on the literature analysis method,this review systematically assessed the effects and mechanisms of Flash-RT and radiotherapy with conventional dose rate on lung injury through searching relevant literatures at home and abroad,so as to provide scientific basis for the treatment of patients with lung cancer by reviewing the comparisons about the effects and mechanisms between Flash-RT and radiotherapy with conventional dose rate on lung injury.Compared with radiotherapy with conventional radiation rate,Flash-RT can significantly reduce lung injury and improve quality of life of patients.It is still demanded to explore the Flash-RT mechanism in future,so as to develop the Flash-RT instrument that is suitable for different tumors and to conduct larger-scale clinical researches.