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 investigate the association between monocyte to high-density lipoprotein cholesterol ratio (MHR) and periodontitis and to provide new epidemiologic evidence on the factors affecting periodontitis. Methods: Data on MHR, periodontitis, and other covariates were selected from the NHANES(National Health and Nutrition Examination) database for 3 cycles of subjects in 2009-2010, 2011-2012, and 2013-2014, and a total of 8 456 study subjects were included. The study participants were grouped according to the prevalence of periodontitis (presence or absence), and three regression models (unadjusted covariates, partially adjusted covariates, and fully adjusted covariates) were constructed to analyze the relationship between MHR and periodontitis by using a weighted logistic regression method with stepwise adjustment for confounders. MHR was divided into four groups from Q1 to Q4 according to quartiles from small to large for weighted trend analysis, and the nonlinear relationship between MHR (continuous) and periodontitis was analyzed using a restricted cubic spline with subgroup analysis and sensitivity analysis. Results: All three logistic regression models showed a positive association between MHR and periodontitis (OR = 2.92, 95%CI: 2.14-3.99, P<0.001 (not adjusted); OR = 1.97, 95%CI: 1.39-2.78, P<0.001 (partially adjusted); OR = 1.62, 95%CI: 1.10-2.39, P = 0.017 (fully adjusted)). Trend analysis showed a significantly higher risk of developing periodontitis in the Q4 group compared with the Q1 group in both single (OR = 1.92, 95% CI: 1.58-2.33, P<0.001) and multifactorial analyses (OR = 1.30, 95% CI: 1.03-1.64, P = 0.029). Restricted cubic spline results did not support a nonlinear relationship between MHR and periodontitis (P for nonlinear>0.05), subgroup analysis showed no significant interaction between the covariates and MHR (P>0.05), and sensitivity analysis also showed a positive correlation between MHR and periodontitis (OR = 1.67, 95%CI: 1.31-2.14, P<0.001). Conclusion MHR is positively associated with the risk of developing periodontitis.

ObjectiveTo investigate the pharmacodynamic effects of Houshihei San （HSHS） recorded with the effects of treating wind and limb heaviness on muscle tissue injury after middle cerebral artery occlusion （MCAO） in rats through the ferroptosis pathway. MethodsThirty SD male rats were selected and randomly grouped as follows： sham， MCAO， deferoxamine mesylate， high-dose HSHS （HSHS-H， 0.54 g·kg^-1）， and low-dose HSHS （HSHS-L， 0.27 g·kg^-1）， with 6 rats in each group. A laser scattering system was used to evaluate the stability of the MCAO model， and rats were administrated with corresponding agents by gavage for 7 days. During the administration period， behavioral， imaging and other methods were used to systematically evaluate the skeletal muscle tissue injury after MCAO and the therapeutic effect in each administration group. Hematoxylin-eosin staining was employed to evaluate the cross-section of muscle cells. Subsequently， immunohistochemistry was used to detect tumor suppressor p53 and glutathione peroxidase 4 （GPX4） in the soleus tissue. Western blot was employed to determine the protein levels of p53， GPX4， myogenic differentiation 1 （MyoD1）， nuclear factor E₂-related factor 2 （Nrf2）， Myostatin， solute carrier family 7 member 11 （SLC7A11）， muscle ring-finger protein-1 （MuRF1）， and muscle atrophy F-box protein （MAFbx） to verify the therapeutic effect in each group. ResultsCompared with the MCAO group， HSHS enhanced the locomotor ability and promoted muscle regeneration， which suggested that the pharmacological effects of HSHS were related to the inhibition of muscle tissue ferroptosis to reduce the expression of muscle atrophy factors. Behavioral and imaging results suggested that compared with the MCAO group， HSHS ameliorated neurological impairments in rats on day 7 （P<0.01）， enhanced 5-min locomotor distance and postural control （P<0.01）， strengthened grasping power and promoted muscle growth （P<0.01）， stabilized skeletal muscle length and weight （P<0.01）， and increased the cross-section of muscle cells （P<0.01）. Compared with the MCAO group， HSHS promoted the increases in glutathione and superoxide dismutase content and inhibited the increase in malondialdehyde content （P<0.05，P<0.01）. Ferroptosis pathway-related assays suggested that HSHS reduced the p53-positive cells and increased the GPX4-positive cells （P<0.01）. HSHS ameliorated muscle function decline after stroke by promoting the expression of GPX4， Nrf2， SLC7A11， and MyoD1 and inhibiting the expression of p53， Myostatin， MurRF1， and MAFbx to reduce ferroptosis in the muscle （P<0.01）. ConclusionHSHS， prepared with reference to the method in the Synopsis of Golden Chamber， can simultaneously reduce the myolysis and increase the protein synthesis in the skeletal muscle tissue after ischemic stroke by regulating the ferroptosis pathway.

Objective: To elucidate the molecular mechanism by which prohibitin 2(PHB2) mediates periodontitis-induced bone tissue inflammation through regulating the nuclear factor kappa B(NF-κB) signaling pathway and its role in irreversible alveolar bone resorption. Methods: Quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR) and immunohistochemistry(IHC) were used to detect the expression differences of inflammatory factors and PHB2 in healthy and inflamed alveolar bone tissues of mice in vivo. In vitro, an inflammatory model was established using lipopolysaccharide(LPS)-induced a mouse calvaria-derived preosteoblastic cell line, subclone E1(MC3T3-E1) cells. Western blot and qRT-PCR were used to clarify the regulatory relationship between PHB2 and inflammatory factors, and immunofluorescence staining was performed to observe changes in PHB2 subcellular localization. PHB2 overexpression plasmids were constructed using molecular cloning, and RNA interference was employed to knock down PHB2 expression to assess its regulatory role in inflammation. Based on RNA-seq data, differential expression analysis based on the negative binomial distribution, version 2(DESeq2) was used for differential expression analysis, and kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment along with gene ontology(GO) functional annotation were performed to identify key signaling pathways and differentially expressed genes. Results: In the mouse periodontitis model, PHB2 expression was significantly upregulated in alveolar bone tissues. In the in vitro inflammatory cell model, PHB2 levels positively correlated with interleukin(IL)-6, IL-1β, and tumor necrosis factor-alpha(TNF-α) levels, and its subcellular localization shifted during inflammation. RNA-seq data and the detection of the level of phosphorylation of p65 protein(p-p65) demonstrated that PHB2 exacerbated inflammatory responses through the NF-κB signaling pathway and was mechanistically linked to upregulation of the upstream chemokine C-X-C motif chemokine ligand 10(CXCL10). Conclusion PHB2 aggravates LPS-induced periodontitis inflammation via the NF-κB signaling pathway, providing new insights into the molecular mechanisms underlying the development of periodontitis.

Nine novel compounds, comprising seven tetrahydroanthraquinones (auxarthrolones A-G, 1-7), a γ-butenolide glycoside (malfilamentoside E, 26), and a γ-butenolide (auxarthrolide A, 27), together with eighteen known compounds (8-25) were isolated from rice-based solid culture of Auxarthron umbrinum (A. umbrinum) DSM3193 using the one strain many compounds (OSMAC) approach. The structural elucidation of these compounds was accomplished through nuclear magnetic resonance (NMR), mass spectrometry (MS), and NMR calculation combined with DP4+ analysis or MAEΔΔδ parameter, while the absolute configurations of new compounds were established through single-crystal X-ray diffraction, electronic circular dichroism (ECD) spectroscopic data analysis and/or chemical derivatization. Austrocortilutein (10) and auxarthrol H (14) demonstrated moderate cytotoxicity against U87 and U251 [half maximal inhibitory concentration (IC₅₀) 3.5-12.1 μmol·L^-1]. Additionally, auxarthrolone A (1), auxarthrol H (14), eupolyphagin B (15), and 7-hydroxy-2-(2-hydroxypropyl)-5-methylchromone (17) exhibited torsional effects on fibroblast proliferation challenges induced by oleic acid, thus demonstrating fibroblast proliferation-promoting activity.
4-Butyrolactone/pharmacology* ; Molecular Structure ; Anthraquinones/pharmacology* ; Humans ; Animals ; Mice ; Cell Line, Tumor ; Magnetic Resonance Spectroscopy

As a medicinal and edible fungus, Inonotus obliquus has a long history of folk application in Russia, Japan, and Northeast China. It is rich in terpenoids, steroids, polysaccharides, phenols, alkaloids, etc, and exhibits pharmacological activities including anti-tumor, hypoglycemic, anti-inflammatory, antioxidant, and lipid-lowering effects. Among these, lanostane-type tetracyclic triterpenes represent its characteristic constituents. This review systematically summarizes the research progress on the chemical components isolated and identified from I. obliquus and their pharmacological activities in recent years. The structures of terpenoids, steroids, and phenolic compounds are compiled and illustrated, with a particular focus on the skeletal types and structural characteristics of lanostane-type tetracyclic triterpenes. This work aims to provide some reference for the further investigation and comprehensive development and utilization of I. obliquus.

Skeletal class Ⅱ malocclusion is characterized by maxillary protrusion,mandibular retrognathia,or a combination of both,and often accompanied by vertical dimensional discrepancies;treatment is complex,and combined orthodontic-orthognathic surgery is needed for the adult patients.Clear aligner therapy has gradually been applied in complex orthodontic cases.However,limited cases have been reported domestically and internationally regarding the application of clear aligner therapy combined with orthognathic surgery.This article presented a case of a patient with skeletal class Ⅱ high-angle malocclusion treated with the combined therapy and analyzed the clinical efficacy of the treatment appraoch to provide reference for the clinical practice.Extraction of impacted wisdom teeth 18,28,38,and 48,as well as orthodontic teeth 15,25,34,and 44,was performed in stages before orthodontic treatment.Clear aligner therapy was used for preoperative orthodontics.In sagittal plane,a super-complete class Ⅱ canine and molar relationship and a 13-14 mm overjet of the anterior teeth were established.The maxillary and mandibular arch morphology was matched horizontally.The orthognathic surgery included maxillary LeFort Ⅰ osteotomy,bilateral sagittal split ramus osteotomy(BSSRO)and chinplasty.Fine occlusal adjustment was conducted after operation.After treatment,the skeletal relationship between upper and lower jaw was corrected to normal;subspinale-nasion-supramental angle(ANB)was improved from 12.3° to 4.7°;the patient established the class Ⅰ canine and molar relationship,with normal overjet and overbite;root parallelism was good and there was no obvious root resorption;the facial soft tissue profile was significantly improved,and nasion-subnasale-pogonion angle(N-Sn-Pg)was improved from 143.9° to 162.8°.The curative effect was stable 1 year after operation.Clear aligner therapy can efficiently complete combined orthodontic and orthodontic surgery in the complex cases.Compared with the fixed appliance,it is more beneficial to the patients'need for beauty and the maintenance of periodontal health.

The purpose of this study was to evaluate the application value of targeted next-generation sequencing(tNGS)for rapid detection of Mycobacterium tuberculosis.From September 2021 to October 2024,1 178 clinical samples from hospitalized pa-tients at the Non-tuberculous Mycobacteria Disease Diagnosis and Treatment Center of Guangzhou Chest Hospital were collected,in-cluding 272 sputum samples,871 broncho alveolar lavage fluid(BALF)samples,and 35 other samples.These samples were analyzed with tNGS and the liquid culture+DNA microarray chip method(referred to as the"culture identification method"),and the detection results between methods were compared.The numbers of Mycobacterium tuberculosis complex(MTBC)samples identified with tNGS and the culture identification method were 172 and 127,respectively,and the numbers of non-tuberculous mycobacteria(NTM)samples detected were 517 and 474,respectively.The detection rate of Mycobacterium was 56.88%with tNGS and was significantly lower(49.49%)with the culture identification method(χ2=13.27,P<0.05).For NTM identification,when sputum was used,the detection rate of tNGS was higher than that of the culture identification method(χ2=24.14,P<0.05).However,when BALF was used,no significant difference in detection rates was observed between tNGS and the culture identification method(χ2=3.97,P=0.06).When different sample types from the same patient were analyzed with tNGS for NTM,BALF was found to be a better choice than sputum(χ2=4.05,P<0.05).However,with the culture identification method,we observed no significant difference between sample types(χ2=2.72,P=0.10).Furthermore,both the MTBC and NTM sequences detected with tNGS were significantly higher in the culture-positive group than the culture-negative group.With increasing sequence numbers,the proportion of cases in the culture-positive group progressively rose,whereas an inverse trend was observed in the culture-negative group.With clinical diagno-sis as the reference standard,the sensitivity,specificity,and Kappa value of tNGS and the culture identification method in identifying mycobacterial diseases were 85.41%,91.32%,0.74 and 74.73%,93.15%,0.63,respectively.Compared with the culture identifica-tion method,tNGS for rapid detection of Mycobacterium exhibited excellent sensitivity,specificity and consistency,and its timeliness should help clinicians make precise individualized treatment plans earlier.

AIM:This study aims to investigate the mechanism by which Zhilianwan(ZLW)decoction affects dextran sodium sulfate(DSS)-induced ulcerative colitis(UC)in mice,focusing on the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway.METHODS:Fifty male C57BL/6J mice were allowed ad libitum access to food for 1 week before the experiment.They were randomly divided into 5 groups:control group,model group,mesalazine(ME)group,high-dose ZLW(ZLW-H)group,and low-dose ZLW(ZLW-L)group,with ten mice in each group.All mice,except those in control group that received distilled water,were given a 3%DSS solution for seven consecutive days,starting the intervention on the first day of modeling.Each group re-ceived their respective treatments via gavage at volume of 20 μL/g for ten consecutive days.The mice in control and model groups received an equivalent volume of distilled water,while those in ME,ZLW-H,and ZLW-L groups were adminis-tered 0.2 g·kg-1·d-1 of ME,10.92 g·kg-1·d-1 of ZLW,and 2.73 g·kg-1·d-1 of ZLW,respectively.Body weight and dis-ease activity index(DAI)scores were recorded daily.At the end of the intervention,colon length was measured,and he-matoxylin-eosin staining was performed to assess pathological changes in colon tissues.ELISA and qPCR were employed to measure serum levels of interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α)and IL-10,as well as the mRNA expression levels in colon tissues.Western blot analysis was conducted to evaluate the protein levels of TLR4,MyD88,cytosolic NF-κB and nuclear NF-κB in colon tissues.RESULTS:Compared with model group,the mice treated with ZLW exhibited an increase in body weight and colon length(P＜0.05),along with a decrease in DAI and colon histo-pathological scores(P＜0.05).Serum levels of IL-1β,IL-6,and TNF-α were significantly lower(P＜0.05),while IL-10 levels increased(P＜0.05).The mRNA expression levels of these inflammatory factors in colonic tissues reflected similar trends.Additionally,levels of TLR4,MyD88,and nuclear NF-κB proteins were reduced(P＜0.05),whereas cytosolic NF-κB protein expression was elevated(P＜0.05)in the colon tissues of treated mice.CONCLUSION:ZLW exerts at-tenuating effects on DSS-induced intestinal injury in UC mice,though modulation of the TLR4/MyD88/NF-κB signaling pathway.

Primary sclerosing cholangitis(PSC)is an autoimmune disease characterized by chronic inflamma-tion and progressive fibrosis that affects both intrahepatic and extrahepatic bile ducts.Despite ongoing research,the under-lying mechanisms of PSC pathogenesis remain incompletely understood.The ductular reaction is not only a key pathologi-cal feature of PSC but also serves as a driving force in its progression.This review examines the promoting effects of the ductular reaction on PSC advancement from multiple perspectives,including the proliferation of biliary epithelial cells,in-flammation,and fibrosis.By providing theoretical insights into the pathogenesis of PSC,this review aims to facilitate the identification of novel therapeutic strategies.