To evaluate the antitumor activity of the novel antibody-drug conjugate (ADC) hIMB1636-MMAE against human epidermal carcinoma, this study systematically investigated its therapeutic efficacy and mechanism of action in A431 cells. Trophoblast cell surface antigen 2 (TROP2) expression in A431 cells was confirmed by Western blot. Flow cytometry was used to analyze the binding characteristics of hIMB1636-MMAE to A431 cells and its effects on cell cycle arrest and apoptosis. The internalization process of the ADC was visualized using confocal laser scanning microscopy, while cytotoxicity in vitro was assessed via the CCK-8 assay. Moreover, an A431 cell-derived xenograft model was established in nude mice to evaluate the in vivo antitumor efficacy of hIMB1636-MMAE. Results showed that A431 cells exhibited high TROP2 expression. hIMB1636-MMAE could specifically bind to A431 cells, and was effectively internalized and transported to lysosomes. Furthermore, hIMB1636-MMAE specifically killed cancer cells with IC₅₀ value of 1.49 ± 0.22 nmol·L^-1, and induced G2/M phase cell cycle arrest and apoptosis. In A431 cell-derived xenografts, hIMB1636-MMAE at 10 mg·kg^-1 exhibited a significant antitumor effect, achieving a tumor inhibition rate of 93.89% (all animal protocols were approved by the Animal Care and Use Committee at the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, No. IMB-20220210D₂03). In conclusion, hIMB1636-MMAE demonstrates promising potential against skin cancer and may serve as a candidate therapeutic for TROP2-positive cutaneous malignancies.

This study aimed to investigate the hepatoprotective effects of the combination of berberine (BBR) and stachyose (Sta) in spontaneous type 2 diabetic KKAy mice. KKAy mice were randomly divided into four groups (n = 14 per group): Model control (model), stachyose (Sta, 200 mg·kg^-1), berberine (BBR, 100 mg·kg^-1), and BBR + Sta (100 mg·kg^-1 BBR + 200 mg·kg^-1 Sta), with oral gavage of water, Sta, BBR, and BBR + Sta respectively for eight weeks. At the end of the experiment, serum and liver tissues were collected, the liver weight was measured, and liver index was calculated. Lipid content and oxidative stress-related indicators in liver were measured. Meanwhile, the levels of lipid, transaminase, and oxidative stress in serum were determined. The expression levels of genes related to lipid metabolism and inflammation in the hepatic tissues were detected using qPCR. The number of macrophages in the liver tissues was measured by flow cytometry. Our findings revealed that the combination of BBR and Sta significantly reduced the liver index, decreased hepatic contents of triglyceride (TG), total cholesterol (TC), nitric oxide (NO), inducible nitric oxide synthase (iNOS), and malonaldehyde (MDA), improved liver histomorphology, attenuated lipid deposition, lowered serum lipids and alanine aminotransferase (ALT) levels, and suppressed the expression of lipid synthesis-related genes [fatty acid synthetase (Fasn), sterol regulatory element-binding protein 1c (Srebp1c)] and inflammation-related genes [interleukin-6 (Il-6), tumor necrosis factor-α (Tnf-α), monocyte chemoattractant protein-1 (Mcp-1)] in hepatic tissues. Furthermore, the combination treatment reduced the total number of hepatic macrophages (CD11b⁺F4/80⁺) and M1 macrophages (F4/80⁺CD11c⁺), and attenuated the antigen-presenting capacity of macrophages. Notably, BBR combined with Sta was superior to BBR alone in reducing blood lipids and inhibiting the expression of hepatic pro-inflammatory genes Il-6. However, no significant difference was observed between the two groups in alleviating hepatic macrophage infiltration and improving oxidative stress indicators. Collectively, the combination of BBR and Sta exerts anti-MASLD (metabolic dysfunction-associated steatotic liver disease) effects by ameliorating hepatic lipid accumulation, inflammation, and oxidative stress, as well as reducing macrophage number and weakening its function. Furthermore, the effects of the combination in improving MASLD are better than BBR alone. These findings suggest that BBR combined with Sta is a potential effective strategy for the treatment of MASLD. The animal experiments were approved by the Institutional Animal Care and Use Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College.

Malassezia furfur (M. furfur) is a common lipophilic fungus with strong epidermal colonization ability and high recurrence rates, capable of inducing a wide range of skin diseases in both animals and humans. Traditional antifungal therapies are associated with significant side effect and frequent drug resistance, necessitating the development of new treatment strategies. This study investigated the efficacy of a protonated phenanthroline-phenanthroline⁺ (ph-ph⁺) compound, previously developed by our team, in inhibiting the growth and reproduction of M. furfur. The results showed that the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of ph-ph⁺ against M. furfur were 2 and 8 µg·mL^-1, respectively. Furthermore, ph-ph⁺ significantly reduced key virulence characteristics of M. furfur, including biofilm formation, hydrophobicity, and adhesion capacity. Assessments using transmission electron microscopy and Annexin V-FITC/propidium iodide staining confirmed that ph-ph⁺ kills M. furfur by inducing apoptosis. In a mouse skin infection model induced by M. furfur, topical application of ph-ph⁺ effectively cleared the fungus from the skin surface without causing skin irritation or other adverse reactions. This study demonstrates that ph-ph⁺ may represent a novel antimicrobial compound distinct from traditional drugs and provides a potential therapeutic approach for treating skin infections caused by M. furfur or other fungi. The experimental protocol was approved by the Animal Experiment Ethics Committee of Southwest University (approval No.: LAC2024-1-0446), and the experiments were conducted strictly in accordance with the ethical principles for the use and care of animals.

This study aims to investigate the effect and mechanism of the chromone compound 19A derived from Saposhnikovia divaricata in ameliorating acute lung injury (ALI) in mice. Molecular docking, drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), and dual-luciferase reporter gene assays were employed to elucidate the binding mode of 19A with farnesoid X receptor (FXR) and its regulatory effect on FXR transcriptional activity. In vitro, an inflammatory model was established by stimulating bone marrow-derived macrophages (BMDMs) with lipopolysaccharide (LPS), and the effects of 19A on cell viability, nitric oxide (NO) release, mRNA expression of inflammatory cytokines, and NF-κB p65 phosphorylation were evaluated. The FXR-dependence of these effects was verified in BMDMs derived from Fxr^-/- mice. In vivo, two ALI mouse models were established via intratracheal instillation of LPS and Escherichia coli (E. coli) to evaluate lung function, total protein concentration in bronchoalveolar lavage fluid (BALF), pulmonary inflammatory histopathology, inflammatory cytokine levels, and reverse validation was performed in Fxr^-/- mice. The use of animals in this study was approved by the Ethics Committee of Shanghai University of Traditional Chinese Medicine, with the approval number: PZSHUTCM2412200007. 19A forms stable interactions with FXR, enhancing its transcriptional activity. In vitro, 19A inhibited LPS-induced NO release and the mRNA expression of inflammatory cytokines, reduced IκBα degradation and phosphorylation, and decreased NF-κB p65 phosphorylation; these anti-inflammatory and NF-κB-inhibitory effects were markedly attenuated or abolished in Fxr^-/- BMDMs. In wild-type mice, 19A markedly improved LPS-induced lung dysfunction, reduced lung index and BALF total protein, alleviated structural damage and inflammatory cell infiltration in lung tissue, and downregulated pulmonary inflammatory cytokine mRNA levels and NF-κB p65 phosphorylation. In contrast, these protective effects of 19A were largely lost in Fxr^-/- mice. In summary, the Saposhnikoviae Radix chromone 19A attenuates LPS-induced macrophage inflammatory responses and ALI in mice by activating FXR and inhibiting the NF-κB signaling pathway. Compound 19A may serve as a lead candidate for ALI prevention and treatment based on the chromone scaffold of Saposhnikoviae Radix, providing experimental support for the modernization of the traditional Chinese medicine Fangfeng and the development of new therapeutics for ALI.

Halorotetin B is a natural terpenoid compound isolated from the tunic of Halocynthia roretzi. Studies have shown that this compound exhibits significant anti-tumor activity. However, its anti-inflammatory potential remains unexplored, likely owing to the multi-target nature of natural compounds. In this study, we investigated the anti-inflammatory activity of Ha. B. The results showed that Ha. B significantly inhibited the production of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-induced RAW264.7 cells. Mechanistically, network pharmacology analysis and Western blot assays verified that Ha. B exerts its anti-inflammatory function by regulating the mitogen-activated protein kinase (MAPK) signaling pathway. Finally, natural product target prediction revealed that Ha. B may directly target mouse double minute 2 (MDM2). Cellular thermal shift assay (CETSA) results confirmed that Ha. B could interact with MDM2 in cells, and molecular docking was performed to simulate their physical binding mode. Meanwhile, RNA interference experiments further verified that Ha. B exerts its anti-inflammatory activity through binding to the E3 ubiquitin ligase MDM2.

Anxiety disorders affect over 3.3% of the global population, with limited treatment options available. Both trace amine-associated receptor 1 (TAAR1) and serotonergic 2C receptor (5-HT_2CR) are implicated in various emotional activities, including anxiety. This study aims to investigate the role of TAAR1/5-HT_2CR in the development of anxiety-like behavior in rats induced by chronic unpredictable mild stress (CUMS), and the intervention effects of a dual agonist. A CUMS-induced rat anxiety model was established. This animal research protocol has been approved by the Yantai University Laboratory Animal Ethics Committee (animal ethics number: YTU20240110). Anxiety-like behavior was assessed using the elevated zero-maze test and open field test. Western blot analysis was employed to detect the expression of TAAR1, 5-HT_2CR, and brain-derived neurotrophic factor (BDNF) proteins in the rat amygdala. To observe the effects of Taar1 and Htr2c gene on anxiety-like behavior in rats, we knocked down or overexpressed these genes in the rat basolateral amygdala (BLA). Results demonstrated that CUMS-induced anxiety significantly reduced the expression levels of TAAR1 and 5-HT_2CR in the amygdala of rats. Overexpression of either Taar1 or Htr2c in the BLA ameliorated anxiety-like behavior. Conversely, knocking down Taar1 or Htr2c in the BLA exacerbated CUMS-induced anxiety-like behavior. Oral administration of PCC0105004 alleviated anxiety-like behavior in CUMS rats, while knocking down both Taar1 and Htr2c in the BLA completely abolished this effect of PCC0105004. Further investigations revealed that PCC0105004 significantly increased the expression of BDNF in the amygdala of rats. These findings indicate that TAAR1 and 5-HT_2CR play crucial roles in the development of anxiety-like behavior in CUMS rats, suggesting that a dual agonist targeting TAAR1/5-HT_2CR may offer a novel therapeutic strategy for anxiety disorders.

The structural and functional homeostasis of the intestinal barrier plays a key role in the occurrence and progression of sepsis. As a highly selective α₂-adrenergic receptor agonist, dexmedetomidine is widely used for sedation in sepsis and perioperative periods requiring surgical intervention. Recent studies have shown that it also exerts anti-inflammatory effects; however, whether it can improve acute intestinal barrier injury in sepsis and the underlying mechanism remain unclear. In this study, an acute sepsis mouse model was established by intraperitoneal injection of lipopolysaccharide (LPS). Evans blue staining, transmission electron microscopy, HE staining, ELISA, and RT-PCR were used to observe the effects of dexmedetomidine (1-6 μg·kg^-1) on intestinal inflammation and barrier function in septic mice. The expression changes of α₂-adrenergic receptors during inflammation, as well as the effects of dexmedetomidine on NF-κB and RhoA/ROCK signaling pathways under LPS stimulation, were further evaluated in mouse intestinal tissues and RAW 264.7 macrophages. The results showed that dexmedetomidine (1-6 μg·kg^-1) administered 6 h after LPS stimulation significantly reduced the levels of inflammatory factors in plasma and intestinal tissues of septic mice, improved intestinal barrier function, and upregulated the expression of tight junction proteins in the small intestine and colon. LPS stimulation downregulated the expression of α₂-adrenergic receptors on macrophages in vivo and in vitro, whereas dexmedetomidine (5-20 nmol·L^-1) reversed this downregulation under inflammatory conditions, inhibited NF-κB pathway activation and inflammatory cytokine secretion in RAW 264.7 cells, and antagonized RhoA/ROCK pathway activation and decreased transepithelial electrical resistance in Caco-2 cells. In conclusion, this study demonstrates that LPS stimulation downregulates α₂-adrenergic receptor expression in macrophages, and dexmedetomidine can upregulate α₂-adrenergic receptors, thereby it may inhibit the NF-κB and RhoA/ROCK pathways, ultimately suppressing inflammation, improving intestinal barrier function, and alleviating acute intestinal barrier injury in sepsis. The animal experiments were approved by the Animal Ethics Committee of Southwest University (Ethics Review No.: IACUC-20260305-12).

This study investigates the protective effects of liquiritin (LQ) against acetaminophen (APAP)-induced liver injury in mice and explores the underlying mechanisms. A drug-induced liver injury model was established in mice by intragastric administration of APAP (300 mg·kg^-1). LQ was administered intragastrically at 1 h after APAP treatment. Serum biochemical indices, oxidative stress markers, and hepatic histopathological changes were evaluated at 6 h after APAP administration to comprehensively assess the ameliorative effects of LQ (animal experiments were approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine, approval Nos. PZSHUTCM2401080003 and PZSHUTCM2510170008). Furthermore, Western blot (WB), quantitative real-time polymerase chain reaction (qPCR), and in vitro cell models were employed to elucidate the underlying molecular mechanisms. The results demonstrated that LQ (25 and 50 mg·kg^-1) significantly ameliorated APAP-induced liver injury (P < 0.05, P < 0.01, P < 0.001), hepatic oxidative stress (P < 0.05, P < 0.01, P < 0.001) and alleviated the inflammatory response (P < 0.05, P < 0.01). WB and qPCR analyses revealed that LQ promoted the nuclear translocation and activation of nuclear factor erythroid 2-related factor 2 (Nrf2), and upregulated downstream gene expression in APAP-induced mouse livers (P < 0.05). Notably, the protective effects of LQ were abolished in Nrf2 knockout mice. These findings indicate that LQ alleviates APAP-induced hepatotoxicity primarily by activating Nrf2 antioxidant signaling pathway to counteract oxidative stress.

Cancer is the second leading cause of death worldwide, with solid tumors accounting for the majority of cases in adults. Advanced and metastatic solid tumors are responsible for most cancer-related deaths; however, current treatments still fail to achieve both high efficacy and low toxicity. In this study, we constructed an oncolytic virus (OV) named SKM_KK010(h) based on chimpanzee adenovirus serotype 23 (SAdV-23). This OV utilizes an optimized human telomerase reverse transcriptase (hTERT) promoter to drive e1a gene expression, thereby restricting viral replication and oncolysis to tumor cells. Additionally, it carries the csf2 transgene, which encodes human granulocyte-macrophage colony-stimulating factor (GM-CSF), to activate systemic anti-tumor immunity. Experiments in vitro demonstrated that SKM_KK010(h) selectively kills a broad range of tumor cell lines — including colon, bladder, liver, lung, and prostate cancer cells — while exhibiting no cytotoxicity toward normal cells. Viral replication in tumor cells was 100- to 1 000-fold higher than in normal cells. Furthermore, SKM_KK010(h) selectively and efficiently expresses bioactive GM-CSF in the tested tumor cells, reaching a level of 147.60 ± 7.63 ng per 10⁶ cells per 24 h (e.g., in LoVo cells), far exceeding the in vivo efficacy threshold (40 ng per 10⁶ cells per 24 h) required to elicit potent antitumor immunity. In contrast, GM-CSF expressions were undetectable or minimal in normal cells. In a LoVo xenograft mouse model, intratumoral injection of SKM_KK010(h) effectively inhibited tumor growth, achieving a relative tumor proliferation rate (T/C%) of 33.01%. In summary, we have successfully developed a safe and effective oncolytic chimpanzee adenovirus with strong potential as a platform for next-generation oncolytic virotherapy. All animal experiments were approved by the Institutional Ethics Committee and conducted in accordance with relevant ethical guidelines (Ethics Approval Number: IACUC-KYB2301P).

The rate constant of the bioorthogonal cleavage reaction is a critical parameter for its in vivo translation and application. Building upon the bioorthogonal decaging chemistry of silylborane and N-oxide, this study conducted structural optimization toward silylborane to obtain derivatives with higher second-order rate constants. The structure-reactivity relationships studies revealed that introducing electron-withdrawing groups onto the phenyl ring of the silylbenzyl moiety significantly enhanced the cleavage reaction rate. Furthermore, several silylboranes were successfully identified with fast reaction kinetics (k₂ > 10² L·mol^-1·s^-1). A representative derivative 2b (k₂ = 239 L·mol^-1·s^-1) was able to efficiently and specifically activate the N-oxide based fluorescent probe and prodrug of tamoxifen at the cellular level. In summary, this work provides an efficient cleavage reaction tool for the design of bioorthogonal prodrugs for in vivo applications.