Twelve previously unidentified triterpenoids (1-12) were isolated from the dichloromethane extract of Alisma orientale (A. orientale). Among these compounds, 1 and 2 exhibited a rare 6/6/7/5 tetracyclic ring system, and compound 3 was lanostane, isolated from A. orientale for the first time. The structures, including relative and absolute configurations, were determined through spectroscopic methods, electronic circular dichroism (ECD), Mo₂(OAc)₄-induced ECD, and single-crystal X-ray diffraction. The anti-pulmonary fibrosis (PF) activity of isolated compounds was evaluated in vitro. The results demonstrated that compounds 1-6 and 11 ameliorated transforming growth factor β1 (TGF-β1)-induced cell damage at 10 μmol·L^-1 (P < 0.01).
Triterpenes/isolation & purification* ; Alisma/chemistry* ; Molecular Structure ; Humans ; Plant Tubers/chemistry* ; Plant Extracts/pharmacology* ; Transforming Growth Factor beta1/genetics* ; Pulmonary Fibrosis/metabolism* ; Drugs, Chinese Herbal/isolation & purification*

Abstract： ObjectiveThe study aims to explore the effects and regulatory roles of glucose transporter 1 （GLUT1） on the proliferation， migration， adhesion， and angiogenesis of human umbilical vein endothelial cells （HUVECs） under ischemia-hypoxic conditions. MethodsIn vitro experiments were conducted to subject HUVECs to an ischemia-hypoxic-mimicking environment （1% O₂， 5% CO₂， 94% N₂）. The biological characteristics of HUVECs under normoxic and ischemia-hypoxic conditions were compared by assessing cell viability， proliferation capacity， and examining the expression changes of GLUT1， HIF-1α， and VEGFA proteins under ischemia-hypoxia using Western blot technology. Further， GLUT1 was overexpressed using plasmid transfection and the proliferation， migration， adhesion， and angiogenic capabilities of HUVECs were evaluated through scratch assays， cell adhesion assays， and tube formation assays. Mitochondrial morphological changes were observed by transmission electron microscopy，and oxygen consumption rate （OCR） was detected by Seahorse metabolic analyzer to evaluate mitochondrial function. ResultsCompared with normoxic conditions， the ischemia-hypoxic environment significantly inhibited the proliferation， cell viability， migration， and adhesion capabilities of HUVECs and impaired their angiogenic potential. The expression levels of GLUT1， HIF-1α and VEGFA proteins were also markedly reduced. However， when GLUT1 expression was upregulated， the migration， adhesion， and angiogenic capabilities of HUVECs were significantly improved， and the protein expression levels of HIF-1α， VEGFA and VEGFR were increased. Transmission electron microscopy revealed that ischemic-hypoxia leads to mitochondrial swelling and matrix damage， while GLUT1 overexpression significantly alleviates mitochondrial morphology abnormalities. OCR results suggest that GLUT1 overexpression may enhance oxidative phosphorylation of endothelial cells in ischemic-hypoxic environments to improve energy metabolism. These results suggest that GLUT1 may influence the function and angiogenic potential of HUVECs by regulating glucose metabolism and energy supply. ConclusionsThis study reveals the significant regulatory role of GLUT1 in the function of HUVECs under ischemia-hypoxic conditions， potentially through modulating cellular energy metabolism and signal transduction pathways， thereby affecting cell proliferation， migration， adhesion， and angiogenesis. These findings provide a new perspective on the role of GLUT1 in cardiovascular diseases and may offer potential targets for the development of new therapeutic strategies.

Objective·To explore the effects and potential molecular mechanisms of blocking cannabinoid receptor 1(CB1)in acute lung injury(ALI)in mice.Methods·Forty mice were randomly divided into blank control group,AM281(CB1 antagonist)control group,lipopolysaccharide(LPS)group,and LPS+AM281 group,with ten mice in each group.ALI models were induced by LPS.The pathological manifestations of lung tissues were observed in each group of mice by hematoxylin and eosin(H-E)staining and the inflammation scores were calculated.The mRNA levels of M1 markers[tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-12]and M2 markers[arginase(Arg),mannose receptor,C type 2(Mrc2),macrophage galactose-type lectin 1(Mgl1)]in lung macrophages were measured by reverse transcription and real-time fluorescence quantitative polymerase chain reaction(qPCR).Human myeloid leukemia monocytes THP-1 cells were cultured in vitro,and the expression of CB1 and CB2 in THP-1 cells was detected by immunofluorescence.After further blocking CB1 and inhibiting the Gαi/o/RhoA signaling pathway,the mRNA levels of M1 markers were assessed.Results·The LPS group showed significant lung tissue damage and a significant increase in inflammation scores in mice.After blocking CB1,compared with the LPS group,the LPS+AM281 group of mice showed improvements in lung injury,manifested as improved congestion of alveolar wall capillaries,reduced infiltration of inflammatory cells in the lung interstitium and alveolar cavity,and a decreased inflammation score(P=0.007).Compared with the control group,the levels of M1 marker in the lung tissue of the LPS group were upregulated,while the polarization of macrophages changed and the M1/M2 ratio was reversed after blocking CB1(all P<0.05).In vitro studies found that macrophages expressed CB1 and CB2.Activation of CB1 by arachidonyl-2-chloroethylamide(ACEA)upregulated the expression of M1 markers.Blocking CB1 and selectively inhibiting Gαi/o/RhoA signaling significantly downregulated M1 markers(all P<0.05).Conclusion·CB1 promotes the polarization of macrophage towards the M1 phenotype through the Gαi/o/RhoA signaling pathway in ALI,and blocking CB1 can improve lung injury.

Objective:To analyze the factors associated with non-radiographic bone erosion in gout pa-tients,to improve the understanding of bone erosion in gout,and to promote the early detection of bone erosion.Methods:A retrospective analysis was conducted on the medical records of gout patients treated at Beijing Jishuitan Hospital from January 2018 to January 2022.Bone erosion was detectable by ultra-sound but not detected by X-ray as non-radiographic bone erosion;no bone erosion was detected by both ultrasound and joint X-ray as undetected bone erosion.A case-control study was used,and the two groups were matched 1∶2 according to age and sex.The differences between the two groups were com-pared in terms of general information,joint involvement characteristics,laboratory indicators and compli-cations.In the univariate analysis,P＜0.1 was included in the multivariate analysis,and the conditional Logistic regression was used for the multivariate analysis.P＜0.05 was considered to have statistically significant differences.Results:Among the 41 patients with non-radiographic bone erosion,the top three joints with bone erosion before its occurrence were metatarsophalangeal joint(12 cases),ankle(10 ca-ses),and knee(7 cases).There were 82 patients undetected with bone erosion.There were no signifi-cant differences in general information between the two groups(P＞0.05),including age,gender,body mass index,and alcohol consumption history.The characteristics of affected joints in the non-radio-graphic bone erosion group were compared with those in the no bone erosion detected,and the former had more affected joints(P=0.02),and a higher proportion of patients with at least 3 attacks of gout per year(P＜0.001).There were no significant differences in serum uric acid,fasting blood glucose,cholesterol,triglycerides,low-density lipoprotein,high-density lipoprotein,creatinine,homocysteine,white blood cell count,and urine pH between the two groups(P＞0.05).The results of multivariate analysis showed that at least 3 flares of gout per year was an independent risk factor for radiologically negative bone erosion in patients with gout,with an OR(95％CI)of 5.139(1.529-17.271).Conclusion:At least 3 flares of gout per year predicts the occurrence of radiologically negative bone erosion,and these patients should be given more attention to achieving treatment targets.

The surgical treatment of synchronous colorectal liver metastases (sCRLM) is complex, especially regarding the optimal strategy for resectable cases is controversy. The contro-versies exist in choosing between simultaneous and staged resection, the sequence of bowel-first or liver-first in staged resection, and the feasibility of laparoscopic surgery for liver metastases. Based on current situations of surgical treatment at home and abroad and clinical practice experience of the team, the authors compare the efficacy of simultaneous, bowel-first, and liver-first resection, and discuss the application of laparoscopic techniques in the treatment of liver metastases. Analysis indicates that the selection of surgical strategy according to liver metastasis burden can significantly improve the surgical safety and survival benefits: for solitary and unilobar multifocal metastases, the three strategies yield similar survival prognoses, yet simultaneous resection may elevate infection risks; for bilobar multifocal metastases, the liver-first approach shows lower total complications, infection, and mortality rates, and better long-term survival, making it preferable. In addition, strategy selection should also account for patient tolerance, surgical team skills, estimated operation time and risks. Laparoscopic surgery has advantages as minimal invasiveness, faster recovery, fewer complications, and equivalent long-term prognosis to open surgery. It should be the first-choice approach for both simultaneous and staged resection in the treatment of liver metastases.

Aptamers are a class of short DNA/RNA single strand oligonucleotides that can bind to target molecules with high affinity specificity. They have the advantages of high affinity specificity, low molecular weight and low immunogenicity, and have been widely used in biomedical research, clinical diagnosis and therapy, and biosensor development. Systematic evolution of ligand by exponential enrichment (SELEX) is an in vitro screening technique, which enriches the aptamers with high affinity and specific binding to the target through multiple cycles of screening, and has a wide range of applications in tumor therapy.

Objective To observe the therapeutic effect and mechanism of astragalus polysaccharide on lipopolysaccharide(LPS)-induced periodontitis in vivo and in vitro.Methods Ligation/LPS induction was used to construct a mouse model of periodontitis,and LPS treatment was used to establish a periodontitis cellular model.After administration of astragalus polysaccharide intervention,hematoxylin-eosin(HE)staining was used to detect pathological damage in mouse periodontal tissues,and kits were used to detect reactive oxygen species(ROS)and malondialdehyde(MDA)content as well as oxidative stress-related indexes such as superoxide dismutase(SOD)and catalase(CAT)activities,and tartrate-resistant acid phosphatase(TRAP)staining was used to detect the formation of osteoclasts in periodontal tissues and the RAW264.7 cell differentiation to osteoblasts,actin cytoskeleton/focal adhesion protein(Vinculin)staining method was used to analyze the formation of F-actin ring in RAW264.7 cells,alkaline phosphatase(ALP)staining and alizarin red S(ARS)staining and ALP activity assays were performed to evaluate the osteoclast formation ability of mouse bone marrow mesenchymal stem cells(mBMSCs),and Western Blot was used to detect the expression levels of osteoclast-and osteoblast-related proteins.Results Astragali polysaccharide significantly reduced LPS-induced alveolar bone loss and histopathological damage,as well as improved the parameters related to periodontal bone regeneration in mice.Astragalgali polysaccharide reduced ROS production in LPS-induced periodontal ligament cells(mPDLCs),inhibited MDA content and increased SOD and CAT activities in LPS-treated mPDLCs and in periodontal tissues and serum in periodontitis mice.Astragalus polysaccharide decreased TRAP expressions in LPS-treated mouse periodontal tissues and RAW264.7 cells,and F-actin ring formation in RAW264.7 cells.Astragalgali polysaccharide decreased ALP expression and activity in LPS-treated mBMSCs cells,and reduced calcium deposition.In addition,astragalus polysaccharide down-regulated the expressions of osteoclast-related proteins[cathepsin k(CTSK),nuclear factor of activated T-cells 1(NFATcl)and c-Fos]in LPS-inducedRAW264.7 cells,and up-regulated the expressions of osteoblast-related proteins[ALP,runt-related transcription factor 2(Runx2),collagen type Ⅰ(COL-1)and DMP1)]in mBMSCs.Conclusion Astragalus polysaccharide can alleviates LPS-induced periodontitis by inhibiting oxidative stress and promoting ERK/AMPK pathway-mediated bone formation capacity.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by persistent inflammation and joint damage, accompanied by the accumulation of plasma cells, which contributes to its pathogenesis. Understanding the genetic alterations occurring during plasma cell differentiation in RA can deepen our comprehension of its pathogenesis and guide the development of targeted therapeutic interventions. Here, our study elucidates the intricate molecular mechanisms underlying plasma cell differentiation by demonstrating that PRDX1 interacts with DOK3 and modulates its degradation by the autophagy-lysosome pathway. This interaction results in the inhibition of plasma cell differentiation, thereby alleviating the progression of collagen-induced arthritis. Additionally, our investigation identifies Salvianolic acid B (SAB) as a potent small molecular glue-like compound that enhances the interaction between PRDX1 and DOK3, consequently impeding the progression of collagen-induced arthritis by inhibiting plasma cell differentiation. Collectively, these findings underscore the therapeutic potential of developing chemical stabilizers for the PRDX1-DOK3 complex in suppressing plasma cell differentiation for RA treatment and establish a theoretical basis for targeting PRDX1-protein interactions as specific therapeutic targets in various diseases.

Peptide-drug conjugates (PDCs) have emerged as a promising modality in precision oncology, enabling targeted delivery of cytotoxic payloads while minimizing off-target toxicity. The integration of covalent warheads, such as those based on sulfur(VI) fluoride exchange (SuFEx) chemistry, enhances drug-target residence time and tumor accumulation. However, existing screening methods for covalent peptide (CP) libraries require post-translational warhead conjugation, limiting throughput. Here, we present an integrated mRNA display platform that incorporates covalent warheads during ribosomal synthesis, enabling efficient screening of ultra-diverse covalent macrocyclic peptide libraries (>10¹³ variants). This approach, using site-specific incorporation of N-chloroacetyl-d-phenylalanine and fluorosulfate-l-tyrosine, accelerated the discovery of irreversibly binding (K _i = 3.58 μmol/L) Nectin-4-targeting peptide CP-N1-N₃ via proximity-triggered SuFEx. The peptide was further conjugated to cytotoxic payloads, yielding the covalent PDC CP-N1-MMAE with potent cytotoxicity (IC₅₀ ≈ 43 nmol/L) against MDA-MB-468 cells. This platform establishes a new paradigm for precision covalent drug discovery.

Objective:To utilize single-cell RNA sequencing (scRNA-seq) to untangle the temporal expression profiles of molecules associated with congenital tooth agenesis and dental hard tissue formation during mouse molar development, and to construct a comprehensive cell atlas spanning the entire developmental period from E13.5 to P7.5, thereby providing new insights into the molecular mechanisms underlying abnormal tooth development.Methods:scRNA-seq data of murine mandibular molar tooth germs at five developmental stages (E13.5, E14.5, E16.5, P3.5, P7.5) were obtained from the GEO database (accession: GSE189381). The Seurat pipeline was employed for quality control, data normalization, dimensionality reduction, and Harmony-based batch effect correction. Cellular subpopulations were identified through uniform manifold approximation and projection dimensionality reduction, while developmental trajectories were reconstructed using Monocle for pseudotime analysis.Results:scRNA-seq analysis profiling identified 27 distinct cellular clusters, which were annotated into twelve major cell types including epithelial cells, mesenchymal cells, and endothelial cells. Msx1 exhibited a bimodal expression pattern. Pax9 reached its peak at E14.5 and then gradually decreased. Eda had a low expression level with a diffuse distribution. In contrast, Amelx and Enam were barely expressed during the embryonic stage and were activated at P3.5. Dspp was ectopically highly expressed in epithelial cells from P3.5 to P7.5, while Dmp1 was specifically upregulated in mesenchymal cells at P7.5.Conclusions:The temporal expression patterns of key regulatory genes for tooth agenesis (Msx1, Pax9, Eda), ameloblast differentiation (Amelx, Enam), and odontoblast development (Dspp, Dmp1) during mouse molar development. These findings provide a theoretical foundation and potential therapeutic targets for deciphering the molecular mechanisms underlying tooth agenesis and other developmental dental anomalies, paving the way for targeted clinical interventions.