Tissue factor (TF), a transmembrane glycoprotein expressed in normal tissues, has a variety of physiological functions in embryonic development, hemostasis and non hemostasis pathways. Studies have found that TF is overexpressed in a variety of tumor tissues and promotes tumor progression. Kaplan Meier (K-M) survival analysis showed that high expression of TF gene was associated with poor prognosis in renal and pancreatic cancer. Therefore, TF has received extensive attention as a target of tumor immunotherapy, and a number of antibody-drug conjugates (ADC) drugs have entered the clinical research stage. In this paper, the gene structure, expression, biological function and the correlation with tumor of TF were systematically elaborated, and the direction of drug design for the new generation of TF-ADC was proposed, in order to provide theoretical support and development direction for the drug research and development of this target.

In recent years, a large number of peptide compounds have been obtained from natural sources or synthesized chemically, which have attracted significant interest due to their high biological activity and low side effects. However, linear peptides encounter many challenges in the field of drug development because they are easily broken down by enzymes and do not pass through cell membranes well. Cyclic peptides, on the other hand, have a stable structure, strong binding to targets, and lower toxicity. They combine the advantages of natural peptides and small molecule drugs in terms of biological activity and drug metabolism, addressing the shortcomings of linear peptides and becoming increasingly important in drug research. This article focuses on the development history of cyclic peptides, discusses the sources, acquisition methods, and specific applications in the field of pharmacology in recent years, and prospects for their future development potential, aiming to provide a theoretical and practical basis for the clinical application of cyclic peptides.

Nutlin-3 is a representative small molecule MDM2-p53 antagonist, which can stabilize the p53 state by disrupting the interaction between p53 and MDM2, thereby inducing the p53 signaling pathway to exert antitumor effects. In this study, six wild-type p53 tumor cell lines, HCT-116, H460, HepG2, MCF-7, A549 and SJSA-1, were used as research objects, and the effects of nutlin-3 on the proliferation of six wild-type p53 cancer cells were detected by methyl thiazolyl tetrazolium (MTT) method and plate cloning assay. The effects of nutlin-3 on H460 cell cycle and apoptosis were detected by flow cytometry. Western blot assay was used to detect ubiquitin-specific protease 7 (USP7), death domain-associated protein (DAXX), murine double minute 2 (MDM2), murine double minute 4 (MDMX/ MDM4), p53, to explore the anti-tumor mechanism of nutlin-3; co-immunoprecipitation (Co-IP) assay was used to detect the effect of nutlin-3 on the interaction between MDM2, MDMX and p53. The results showed that nutlin-3 inhibited the proliferation of H460 in a time- and concentration-dependent manner. The results of cell cycle and apoptosis showed that nutlin-3 could block the H460 cell cycle in the G0/G1 phase, and induce apoptosis by activating cleaved-PARP. Western blot results showed that nutlin-3 could up-regulate the expression of USP7, DAXX, MDM2, MDMX and p53 in H460 cells. Co-IP results showed that nutlin-3 inhibited the protein interactions between MDM2 and p53 and MDM2 and MDMX. In conclusion, nutlin-3 can significantly inhibit the proliferation of wild-type p53 cancer cells and induce cell cycle arrest and apoptosis, which may be related to the disruption of MDM2/MDMX's interaction with p53 to activate the MDM2-p53 signaling pathway.

In the early stage of the project, it was found that natural pentacyclic triterpenes liquidambaric acid regulates the NEDD8 modification of Cullin2. This study aims to find more triterpenoid natural active molecules targeting Cullin family members and reveal its mechanism of action. Western blot was used to detect natural products that can significantly change the total protein NEDD8 modification and specific Cullin protein NEDD8 modification in cells; microscale thermophoresis (MST) was used to detect the direct binding of candidate small molecule oleanonic acid to TRAF family proteins, and the binding at the level of living cells was verified by cellular thermal shift assay (CETSA). Proximity ligation assay (PLA) was used to investigate the regulatory effect of oleanonic acid on the protein interaction between TNF receptor-associated factor 1 (TRAF1) and Cullin1 NEDD8 modified complex. Three pentacyclic triterpenoids were found to significantly inhibit NEDD8 modification in cells, among which oleanonic acid had the strongest effect on blocking NEDD8 modification. Different from the previous identification that liquidambaric acid regulates Cullin2/5, oleanonic acid can also specifically induce NEDD8-modified Cullin1 to transform into its unmodified form. And binding experiments showed that oleanonic acid could directly bind to TRAF1 at the level of cell lysate and living cells. Further mechanism studies found that oleanonic acid significantly changed the protein interaction between TRAF1 and Cullin1 NEDD8 modified complex. The above results indicate that oleanonic acid targets TRAF1 and regulates its interaction with NEDD8 modification complex to inhibit NEDD8 modification of Cullin.

2',4'-Dimethoxychalcone (DMC) is a structural modifier of carvacrol B. In this study, gastric cancer cells MGC-803 and HGC-27 were used as the subjects to investigate the anti-tumor effect and mechanism of DMC on gastric cancer (GC) cells both in vitro and in vivo. DMC inhibited cell viability and cell proliferation and promoted cell apoptosis in GC cells, detected by CCK-8 assay, EdU staining and Annexin V-FITC/PI double-staining flow cytometry. A nude mouse model of GC cell xenograft was constructed by subcutaneous injection with MGC-803 cells, for measuring the effect of DMC on the growth of GC in vivo, and DMC inhibited the growth of subcutaneous transplantation tumor in nude mice. The animal experiments were approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine under the ethical number PZSHUTCM2310110002. The effect of DMC on the RNA expression of MGC-803 cells was detected by RNA-seq assay, and it was found that the biological function of DMC was enriched in glycolysis. DMC inhibited the glucose uptake capacity and lactate production and efflux of gastric cancer cells, detected by using 2-NBDG probe with flow cytometry and lactate (LD) test kit. Western blot assay was performed to detect the protein expression of proliferation, apoptosis, and glycolysis-related proteins in gastric cancer cells, and the results demonstrated that DMC up-regulated the protein expression of cleaved caspase-9, cleaved caspase-3, cleaved PARP, down-regulated Ki-67 protein expression, and inhibited the protein expression of c-Myc, LDHA, GLUT3, PDHK1 and MCT1 in gastric cancer cells. The Seahorse energy metabolism analyser was used to measure the rate of glycolysis, and it was found that DMC could down-regulate the basal glycolysis rate and compensatory glycolysis in gastric cancer cells. The c-Myc overexpressing cell line MGC-803 was used in the reversal experiment to further confirm that DMC suppressed gastric cancer growth through inhibiting c-Myc mediated glucose uptake and glycolysis. In conclusion, DMC may inhibit the protein expression of c-Myc and its target glycolysis-related genes, suppressed c-Myc-mediated glucose uptake and glycolysis in gastric cancer cells, thereby inhibited the cell proliferation and promoted cell apoptosis of gastric cancer cells, and thus finally inhibited the growth of gastric cancer in vivo and in vitro.

Oxaliplatin (Oxa) is a chemotherapy drug commonly used for advanced colorectal cancer, however most patients develop resistance after treatment while the mechanisms of which have not been fully elucidated. In this study, oxaliplatin resistant cell lines were constructed from human colorectal cancer HCT116 cells through concentration gradient induction. On this basis, we investigated the expression profiling of HCT116/Oxa cells based on quantitative proteomics. Gene ontology (GO) analysis was conducted via The Database for Annotation, Visualization, and Integrated Discovery Database (DAVID), and pathway enrichment analysis was done using GeneAnalytics database. The potential targets and molecular mechanisms of oxaliplatin resistance in colorectal cancer were further studied by inhibitors, Western blot and siRNA. The results showed that the oxaliplatin resistance index of HCT116/Oxa cells was 10.2. HCT116/Oxa cells demonstrated stronger proliferation potential and anti-apoptotic capacity to oxaliplatin compared with HCT116 cells. Proteomic data demonstrated significant expression change of 717 genes in HCT116/Oxa cells, among which 399 genes were up-regulated while 318 ones down-regulated comparing with HCT116 cells. GO enrichment analysis showed that differentially expressed genes were mainly related to biological processes such as oxidative stress response, iron metabolism, lipid metabolism, apoptosis and cell cycle progression. Pathway analysis displayed notable changes of cell metabolism, ferroptosis, Nrf2-ARE signaling, fatty acid and glutathione metabolism in HCT116/Oxa cells. Quantitative results indicated that the expression of proteins directly related to ferroptosis, including glutathione peroxidase 4 (GPX4), glutamate-cysteine ligase regulatory subunit (GCLM), ferritin light chain (FTL), ferritin heavy chain (FTH1), heme oxygenase 1 (HMOX1), glutathione reductase (GSR) and NADH dehydrogenase 1 (NQO1) increased, while long chain fatty acid-CoA ligase (ACSL) 4 and ACSL1 decreased significantly in HCT116/Oxa cells. Functional studies showed that RSL3, a specific inhibitor of GPX4, decreased the viability of drug-resistant cells, improved lipid peroxidation, increased the concentration of ferrous ions, malondialdehyde, and decreased the concentration of glutathione (GSH). Western blot showed that the expressions of GPX4, FTH1, FTL and GSR increased in HCT116/Oxa, while ACSL4 decreased. RSL3 reversed the levels of GPX4, FTH1, FTL, GSR and ACSL4. It was further found that knockdown of GPX4 decreased the viability of drug-resistant cells, increased lipid peroxidation levels and decreased GSH concentration. These results suggest that ferroptosis resistance mediated by GSH/GPX4 pathway may be a potential mechanism of oxaliplatin resistance in HCT116/Oxa, and inhibition of GSH/GPX4 signaling could be an effective approach to reverse oxaliplatin resistance in colorectal cancer.

Polydatin (PD) is a natural active crystalline compound extracted from the roots and stems of Polygonum cuspidatum, and is a natural precursor of resveratrol. This study aims to investigate the therapeutic effects of PD on monosodium urate (MSU)-induced gouty arthritis in mice and its potential mechanisms. The animal experiment has been approved by the Ethics Committee of Nanjing University (approval number: 2407002). A gouty arthritis model was established by injecting 20 μL of MSU (25 mg·mL^-1) suspension into the mouse plantar. The effect of PD on pathological changes in the mouse plantar was evaluated. The treatment group received daily intraperitoneal injections of different doses of PD (low dose: 5 mg·kg^-1, medium dose: 10 mg·kg^-1, high dose: 20 mg·kg^-1) for 3 days before model induction. The thickness of the mouse plantar was measured and photographed at 3, 6, 9, 12, and 24 h after MSU suspension injection. Histopathological damage to the plantar tissue was observed using hematoxylin-eosin (H&E) staining. Immunohistochemistry and immunofluorescence were used to detect the expression of NLRP3 and CASP1 p20 to assess NLRP3 inflammasome activation in the plantar tissue. At the cellular level, lipopolysaccharide (LPS) combined with adenosine triphosphate (ATP)/MSU/nigericin was used to construct a cellular activation model of the NLRP3 inflammasome. ELISA was used to detect the effect of PD on interleukin-1β (IL-1β) secretion after NLRP3 inflammasome activation in macrophages. Flow cytometry was employed to measure CASP1 p20 activation in macrophages. Immunofluorescence was used to examine NLRP3 inflammasome assembly in macrophages. The results of the study indicate that, compared to the model group, the PD-treated group exhibited a significant reduction in the swelling of the mouse plantar. H&E staining showed a notable reduction in tissue damage in the mouse plantar, suggesting that PD has a therapeutic effect on plantar damage in mice. Immunohistochemistry and immunofluorescence results revealed a significant decrease in the expression of CASP1 p20 and NLRP3, indicating that PD significantly inhibits the activation of the NLRP3 inflammasome, thereby attenuating the local inflammatory response in the mouse plantar. At the cellular level, PD treatment significantly reduced the secretion of IL-1β and activation of CASP1 p20, both of which are mediated by NLRP3 inflammasome activation. Furthermore, NLRP3 inflammasome assembly was inhibited. In summary, PD exerts its anti-inflammatory effect by suppressing the assembly and activation of the NLRP3 inflammasome, reducing the production and release of the pro-inflammatory cytokine IL-1β, thereby alleviating joint damage in mouse gouty arthritis. This provides a novel strategy for the treatment of gout.

Liver fibrosis is a chronic liver injury caused by various pathogenic factors, leading to excessive accumulation of extracellular matrix such as collagen. It represents a common pathological hallmark during the progression of most chronic liver diseases. However, there is currently no universally recognized specific and effective drug for the clinical treatment of liver fibrosis. Therefore, this study investigates the effects of Alisma Rhizoma on bile duct ligation (BDL)-induced liver fibrosis and explores the potential pharmacological mechanisms. The animal experimental protocol was reviewed and approved by the Animal Welfare and Ethics Committee of Shanghai University of Traditional Chinese Medicine (registration No. PZSHUTCM2303280007), in compliance with relevant animal welfare and ethical standards. Mice were subjected to BDL to induce liver fibrosis. Mice were divided into five groups: sham operation group (Sham), model group (BDL), ethanol extract protection group (BDL+EE, 1.6 g·kg^-1), water extract protection group (BDL+WE, 4.0 g·kg^-1), obeticholic acid protection group (BDL+OCA, 10 mg·kg^-1). The results showed that both of EE and WE could attenuate BDL-induced liver fibrosis as evident by reduced serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT) activities, total bile acids (TBA) levels, and improved pathological conditions such as cholestasis, collagen deposition, inflammatory cell infiltration, and liver tissue necrosis. Notably, EE showed better efficacy than WE. Further studies showed that EE improved liver fibrosis dose dependently. EE treatment impaired the bile acids homeostasis in serum and liver, and recovered the hepatic mRNA expression of farnesoid X receptor (FXR) as well as the downstream genes including small heterodimer partner (SHP), cholesterol 7-alpha hydroxylase (CYP7A1) and bile salt export pump (BSEP). Further study also proved that the four major triterpenes in EE increased the transcriptional activities of FXR in vitro. This study provides a theoretical basis for the clinical application of Alisma Rhizoma in the prevention and treatment of liver fibrosis.

Fusobacterium nucleatum (Fn) is closely associated with the occurrence and progression of colorectal cancer (CRC). The development of specific antibacterial agents targeting Fn is crucial for the prevention and treatment of CRC. Based on the preliminary phenotypic screening results from our research group, dimetridazole was successfully identified as a hit compound with antibacterial activity against Fn. In this preliminary structural optimization study, we designed and synthesized seven novel nitroimidazole derivatives comprising three structural types, followed by antimicrobial evaluation of all target compounds. Among them, compound CL6 exhibited excellent antibacterial activity against Fn (MIC = 0.5 μg·mL^-1) and demonstrated good selectivity towards intestinal bacteria and normal cells. Compound CL6 significantly inhibited the migration of CRC cells (HCT116) induced by Fn preliminary mechanistic studies suggest that compound CL6 disrupts the integrity of the Fn bacterial biofilm and cell wall, providing a promising lead compound for the development of novel anti-Fn drugs.

The 95% ethanol extract of Curcuma longa was isolated and purified by silica gel column chromatography, polyamide column chromatography, preparative thin layer chromatography and semi-preparative HPLC. Then, the structures of the obtained compounds were identified by HR-MS, IR, and NMR. Absolute configuration of the new compound was determined by calculating ECD. Finally, two bisabolane-type sesquiterpenoids were obtained from C. longa and identified as (7S)-1,3,5,10-bisabolatetraen-3-nitro-9-one (1) and turmeronol A (2). Compound 1 was a novel nitro-substituted bisabolane-type sesquiterpenoid.