The main protease （M^pro） of SARS-CoV-2, a pivotal enzyme for viral replication and transcription, has emerged as a critical therapeutic target in antiviral drug discovery due to its high conservation across coronaviruses and low homology with host proteases. Recent advances in M^pro inhibitors were summarized in this review, including peptide-mimetic covalent inhibitors, non-peptide covalent inhibitors, and non-covalent inhibitors. Furthermore, the application of proteolysis targeting chimera （PROTAC） technology in developing M^pro degraders was explored, which provided valuable insights for the development of antiviral drugs.

Background China is a major coal producer and consumer country in the world. Coal workers' pneumoconiosis (CWP) is a primary factor endangering the occupational health of coal miners. Research on the disease burden of CWP and its changing trend is significant for disease prevention & control and associated policies. Objective To analyze the disease burden of CWP in China from 1990 to 2021 and its changing trend, and predict the disease burden from 2022 to 2035. Methods Using the Global Burden of Disease Study (GBD) database of 2021, numbers ofincident cases, prevalent cases, deaths, and disability-adjusted life years (DALYs) as well as crude and age-standardized rates of CWP in China were retrieved. Linear regression model was used to calculate the estimated annual percentage change (EAPC) of the age-standardized rates. Joinpoint regression model was used to analyze the temporal trend of disease burden and the disease burden of different sexes and age groups, and Bayesian age-period-cohort (BAPC) model was used to forecast the trend of CWP disease burden. Results In 1990, the incident, prevalent, and deaths cases of CWP in China were 3266, 18872, and 1561, respectively, and the DALYs of CWP were 44614.718 person-years. In 2021, the incident, prevalent, and deaths cases of CWP were 3446, 23975, and 1229, respectively, and the DALYs of CWP were 29610.754 person-years. From 1990 to 2021, the age-standardized incidence, prevalence, mortality, and DALYs rates of CWP in China all showed a downward trend, with the EAPCs of −3.121%, −2.532%, −4.018%, and −4.268%, respectively. The disease burden of CWP in China was mainly concentrated in the male population. The annual average percent change analysis indicated that each standardized rate showed a fluctuating downward trend in different periods. The BAPC model showed that from 2022 to 2035, the age-standardized rates of CWP in China would continue to decline steadily. In 2035, the age-standardized incidence, prevalence, mortality, and DALYs rates of CWP would be reduced to 0.10 per 100000, 0.74 per 100000, 0.03 per 100000, and 0.74 per 100000, respectively. Conclusion The disease burden contributed by CWP in China generally show a downward trend from 1990 to 2021, and it is expected that the age-standardized rates will continue to decline steadily from 2022 to 2035.

BACKGROUND:The formation of postoperative abdominal adhesions is a complicated process,and the prevention of postoperative adhesions is an urgent problem in clinic. OBJECTIVE:To analyze the mechanism of adhesion at cellular and molecular levels,and to provide theoretical basis for the prevention and treatment of adhesion by mesenchymal stem cell exosomes. METHODS:"Abdominal adhesion,pelvic adhesion,postoperative adhesion,epithelial mesenchymal transformation,mesenchymal stem cells,stem cell exosomes,mesenchymal stem cell exosomes"were selected as Chinese and English search terms.We searched PubMed,CNKI,and Chinese biomedical literature and screened relevant articles on postoperative abdominal adhesion and mesenchymal stem cell exosomal intervention published from inception to August 2023.After systematic analysis,54 articles were finally included for the review. RESULTS AND CONCLUSION:(1)Any pathological factors such as peritoneal inflammation,mechanical injury,tissue ischemia,and foreign body implantation cause peritoneal surface injury,resulting in postoperative abdominal adhesion.The formation process of adhesion includes the interaction of peritoneal mesothelial cell repair,inflammatory response,fibrinolytic system,coagulation pathway and other processes,involving a variety of cytokines and signaling pathways.Wnt/β-catenin pathway can induce fibrosis and angiogenesis,and cooperate with transforming growth factor-β/Smads signaling pathway to stimulate fibroblast proliferation and cause peritoneal fibrosis.Meanwhile,nuclear factor-κB signaling pathway up-regulates the expression of cellular inflammatory factors,promotes fibroblast proliferation,and plays a key role in the process of tissue fibrosis.(2)The paracrine function of stem cells is an important direction of molecular intervention in abdominal adhesions based on regenerative medicine.It can participate in a variety of complex cytokines and signaling pathways involved in abdominal adhesions.(3)Compared with traditional methods for treating abdominal adhesions,mesenchymal stem cell exosome has biological activity and is safe to use.Mesenchymal stem cell exosomes without special culture and expansion have lower immunogenicity,longer stability and other advantages,can guide a normal repair and healing through a variety of ways.(4)Mesenchymal stem cell exosome has been proven to be involved in regulating the above processes of adhesion formation in previous studies,showing potential application prospects in clinical studies.However,further clinical studies are needed to explore appropriate treatment options for mesenchymal stem cell exosomes to address the problem of clinical translation.

Objective To design and synthesize PROTAC degraders targeting the SARS-CoV-2 main protease （M^pro）based on PROTAC technology. Methods Compound 3w was used as the M^pro ligand, and the indole N atom in the solvent-exposed region was selected as the linker attachment site. A series of M^pro PROTACs were designed and synthesized by conjugating compound 3w with the CRBN ligand pomalidomide through alkane linkers of different lengths. The structures of the target compounds were confirmed by ¹H NMR, ¹³C NMR, and HRMS. Western Blot analysis was employed to evaluate their degradation activity and explore its mechanism in M^pro-HEK-293T cells. Results Four novel M^pro PROTACs（A1-A4）were successfully synthesized. The most potent compound A4 demonstrated M^pro degradation activity with a DC₅₀ value of 5.2 μmol/L, and its degradation mechanism was validated. Conclusion A novel class of M^pro PROTAC degraders were successfully designed and synthesized, and their protein degradation capability and mechanism of action were demonstrated. These results provided lead compounds for the research and development of antiviral degraders against SARS-CoV-2.

Purpose To explore the role and mechanism by which the forkhead box C1(FOXC1)promoter up-stream transcript(FOXCUT)regulates proliferation and invasion of non-small cell lung carcinoma(NSCLC)cells.Methods Bioinformatic analysis and RT-qPCR were used to quantify FOXCUT expression in NSCLC tissues.After FOXCUT knockdown in NSCLC cell lines,cell proliferation was examined using CCK-8 and EdU assays,and invasion was evaluated by Transwell assay.The expression of E-cadherin,vimentin,N-cadherin,and FOXC1 was detected by Western blot.FOXCUT-silenced H460 cells were constructed using lentiviruses and subcutaneously injected into nude mice to observe tumor growth.To rescue FOXC1 expression,an FOXC1 expression plasmid was transfected into FOX-CUT-knockdown cells.LncBook 2.0,ENCORI,and TargetScan databases were queried to predict miRNAs that inter-act with FOXCUT and FOXC1.Results FOXCUT expression was significantly higher in NSCLC tissues than in normal lung tissues(normal:0.24±0.22 vs NSCLC:0.68±0.76,t=5.94,P＜0.001),and patients with high FOXCUT expression had a poorer prognosis(P＜0.01).FOXCUT interference markedly repressed NSCLC cells' proliferation and invasion(P＜0.01).FOXCUT knockdown significantly upregulated E-cadherin and downregulated vimentin and N-cadherin(P＜0.01).In vivo,FOXCUT-silenced cells formed significantly smaller tumors in nude mice(P＜0.01).FOXCUT knockdown markedly reduced FOXC1 expression(P＜0.01).Overexpression of FOXC1 in FOX-CUT-depleted cells rescued cell proliferation(P＜0.01).Bioinformatic analysis identified 8 miRNAs potentially co-regulated by FOXCUT and FOXC1.Conclusion Knockdown of FOXCUT restrains NSCLC cell proliferation and inva-sion,possibly through suppression of FOXC1 expression.

Membrane protein degradation is a cutting-edge field in targeted protein degradation (TPD). Herein, we developed glypican-3 (GPC3)-mediated lysosome-targeting chimeras (GLTACs) as a novel strategy for the targeted degradation of tumor-specific membrane proteins. GLTACs utilize tumor-specific expression and endocytosis properties of GPC3 to degrade membrane proteins. By conjugating a GPC3-targeting peptide with the ligand of protein of interest (POI), GLTACs induce the formation of a ternary complex that is internalized into lysosomes, leading to the degradation of the POI. The effectiveness and specificity of GLTACs were validated by designing PD-L1, c-Met, and FGFR1 degraders. In particular, GLTAC WP0 potently degraded PD-L1 and induced T-cell-mediated tumor killing against HepG2 cells, highlighting the potential therapeutic applications. The development of GLTAC technology expands the scope of TPD strategies and opens new avenues for discovering novel therapeutic modalities against challenging protein targets.

Purpose To explore the role and mechanism by which the forkhead box C1(FOXC1)promoter up-stream transcript(FOXCUT)regulates proliferation and invasion of non-small cell lung carcinoma(NSCLC)cells.Methods Bioinformatic analysis and RT-qPCR were used to quantify FOXCUT expression in NSCLC tissues.After FOXCUT knockdown in NSCLC cell lines,cell proliferation was examined using CCK-8 and EdU assays,and invasion was evaluated by Transwell assay.The expression of E-cadherin,vimentin,N-cadherin,and FOXC1 was detected by Western blot.FOXCUT-silenced H460 cells were constructed using lentiviruses and subcutaneously injected into nude mice to observe tumor growth.To rescue FOXC1 expression,an FOXC1 expression plasmid was transfected into FOX-CUT-knockdown cells.LncBook 2.0,ENCORI,and TargetScan databases were queried to predict miRNAs that inter-act with FOXCUT and FOXC1.Results FOXCUT expression was significantly higher in NSCLC tissues than in normal lung tissues(normal:0.24±0.22 vs NSCLC:0.68±0.76,t=5.94,P＜0.001),and patients with high FOXCUT expression had a poorer prognosis(P＜0.01).FOXCUT interference markedly repressed NSCLC cells' proliferation and invasion(P＜0.01).FOXCUT knockdown significantly upregulated E-cadherin and downregulated vimentin and N-cadherin(P＜0.01).In vivo,FOXCUT-silenced cells formed significantly smaller tumors in nude mice(P＜0.01).FOXCUT knockdown markedly reduced FOXC1 expression(P＜0.01).Overexpression of FOXC1 in FOX-CUT-depleted cells rescued cell proliferation(P＜0.01).Bioinformatic analysis identified 8 miRNAs potentially co-regulated by FOXCUT and FOXC1.Conclusion Knockdown of FOXCUT restrains NSCLC cell proliferation and inva-sion,possibly through suppression of FOXC1 expression.

A LAMP-Taqman rapid detection system for porcine pseudorabies virus(PRV)was de-veloped based on LAMP and quantitative PCR.LAMP primers were designed for PRV conserved sequences,and the loop primer modified by the fluorescent quenching group was used as the Taq-man probe.The composition optimization,specificity,sensitivity and repeatability of the LAMP-Taqman system were tested using positive samples and recombinant plasmid as templates.Thirty-eight samples of pork swabs were tested with the commercial LAMP detection kit in parallel to verify the actual detection effect of the LAMP-Taqman detection system.The results showed that the optimal final concentration of each component was as follows:PRV-FIP/BIP 0.8 μmol/L,Bst DNA polymerase 0.7 U/μL,Taq DNA polymerase 0.24 U/μL,dNTPs 1.6 mmol/L,MgSO47.2 mmol/L.This system had good specificity and did not cross-react with other virus samples.The linear correlation coefficient of gradient samples was 0.995,the coefficient of variation of repeatable tests was less than 3.000％,and the minimum detection limit could reach 2.81 ×102 copies/μL.The test results of the actual swab samples were consistent with the commercial isothermal fluores-cence detection reagents.In conclusion,the PRV detection system established by LAMP-Taqman method in this study is specific,sensitive,stable and accurate,and is a reliable technical method suitable for the accurate detection of porcine PRV.

Liver cancer is the second leading cause of cancer-related deaths worldwide, with hepatocellular carcinoma (HCC) being the most common form of primary liver cancer. Glypican-3 (GPC3) is a cell membrane proteoglycan which is rarely expressed in normal adult tissues but is specifically upregulated in HCC, which makes GPC3 a reliable target for the diagnosis and treatment of HCC. The role of GPC3 in the regulation of cancer development through Wnt, YAP, hedgehog and other signaling pathways were reviewed in this article. GPC3-targeted therapies, such as monoclonal antibodies, bispecific antibodies, tumor vaccines, immunotoxins, CAR-T cells, and photosensitizer therapy were also summarized. These treatment methods offered promising approaches for HCC treatment and future treatment strategies for HCC based on GPC3 were prospected in this paper.

Linear ubiquitination is an important post-translational modification that has been discovered in recent years. The linear ubiquitin chain is formed by the linkage of glycine residue of one ubiquitin protein to the methionine residue of another ubiquitin. This process is regulated by the linear ubiquitin chain assembly complex (LUBAC) and the OTU deubiquitinase with linear linkage specificity (OTULIN). Linear ubiquitination is involved in various biological processes, including immune response, inflammation, and cell apoptosis. Recent studies have shown that linear ubiquitination is closely related to the occurrence, development, and drug resistance of tumors by affecting signaling pathways such as NF-κB and Wnt/β-catenin. The research progress on the function of LUBAC and OTULIN in tumors was reviewed in this paper.