Bioorthogonal labeling and profiling of N6-isopentenyladenosine (i6A) modified RNA.

Chemical modifications in RNAs play crucial roles in diversifying their structures and regulating numerous biochemical processes. Since the 1990s, several hydrophobic prenyl-modifications have been discovered in various RNAs. Prenyl groups serve as precursors for terpenes and many other biological molecules. The processes of prenylation in different macromolecules have been extensively studied. We introduce here a novel chemical biology toolkit that not only labels i6A, a prenyl-modified RNA residue, by leveraging the unique reactivity of the prenyl group, but also provides a general strategy to incorporate fluorescence functionalities into RNAs for molecular tracking purposes. Our findings revealed that iodine-mediated cyclization reactions of the prenyl group occur rapidly, transforming i6A from a hydrogen-bond acceptor to a donor. Based on this reactivity, we developed an Iodine-Mediated Cyclization and Reverse Transcription (IMCRT) tRNA-seq method, which can profile all nine endogenous tRNAs containing i6A residues in Saccharomyces cerevisiae with single-base resolution. Furthermore, under stress conditions, we observed a decline in i6A levels in budding yeast, accompanied by significant decrease of mutation rate at A37 position. Thus, the IMCRT tRNA-seq method not only permits semi-quantification of i6A levels in tRNAs but also holds potential for transcriptome-wide detection and analysis of various RNA species containing i6A modifications.

Integrative analysis of single-cell embryo data reveals transcriptome signatures for the human pre-implantation inner cell mass.

As the source of embryonic stem cells (ESCs), inner cell mass (ICM) can form all tissues of the embryo proper, however, its role in early human lineage specification remains controversial. Although a stepwise differentiation model has been proposed suggesting the existence of ICM as a distinct developmental stage, the underlying molecular mechanism remains unclear. In the present study, we perform an integrated analysis on the public human preimplantation embryonic single-cell transcriptomic data and apply a trajectory inference algorithm to measure the cell plasticity. In our results, ICM population can be clearly discriminated on the dimension-reduced graph and confirmed by compelling evidences, thus validating the two-step hypothesis of lineage commitment. According to the branch probabilities and differentiation potential, we determine the precise time points for two lineage segregations. Further analysis on gene expression dynamics and regulatory network indicates that transcription factors including GSC, PRDM1, and SPIC may underlie the decisions of ICM fate. In addition, new human ICM marker genes, such as EPHA4 and CCR8 are discovered and validated by immunofluorescence. Given the potential clinical applications of ESCs, our analysis provides a further understanding of human ICM cells and facilitates the exploration of more unique characteristics in early human development.

Advanced glycation end product-modified low-density lipoprotein promotes pro-osteogenic reprogramming via RAGE/NF-κB pathway and exaggerates aortic valve calcification in hamsters.

BACKGROUND: Advanced glycation end product-modified low-density lipoprotein (AGE-LDL) is related to inflammation and the development of atherosclerosis. Additionally, it has been demonstrated that receptor for advanced glycation end products (RAGE) has a role in the condition known as calcific aortic valve disease (CAVD). Here, we hypothesized that the AGE-LDL/RAGE axis could also be involved in the pathophysiological mechanism of CAVD. METHODS: Human aortic valve interstitial cells (HAVICs) were stimulated with AGE-LDL following pre-treatment with or without interleukin 37 (IL-37). Low-density lipoprotein receptor deletion (Ldlr-/-) hamsters were randomly allocated to chow diet (CD) group and high carbohydrate and high fat diet (HCHFD) group. RESULTS: AGE-LDL levels were significantly elevated in patients with CAVD and in a hamster model of aortic valve calcification. Our in vitro data further demonstrated that AGE-LDL augmented the expression of intercellular cell adhesion molecule-1 (ICAM-1), interleukin-6 (IL-6) and alkaline phosphatase (ALP) in a dose-dependent manner through NF-κB activation, which was attenuated by nuclear factor kappa-B (NF-κB) inhibitor Bay11-7082. The expression of RAGE was augmented in calcified aortic valves, and knockdown of RAGE in HAVICs attenuated the AGE-LDL-induced inflammatory and osteogenic responses as well as NF-κB activation. IL-37 suppressed inflammatory and osteogenic responses and NF-κB activation in HAVICs. The vivo experiment also demonstrate that supplementation with IL-37 inhibited valvular inflammatory response and thereby suppressed valvular osteogenic activities. CONCLUSIONS: AGE-LDL promoted inflammatory responses and osteogenic differentiation through RAGE/NF-κB pathway in vitro and aortic valve lesions in vivo. IL-37 suppressed the AGE-LDL-induced inflammatory and osteogenic responses in vitro and attenuated aortic valve lesions in a hamster model of CAVD.

Detecting 2'-5'-adenosine linked nucleic acids via acylation of secondary hydroxy functionality.

2'-5'-Adenosine linked nucleic acids are crucial components in living cells that play significant roles, including participating in antiviral defense mechanisms by facilitating the breakdown of viral genetic material. In this report, we present a chemical derivatization method employing 5-fluoro-2-pyridinoyl-imidazole as the acylation agent, a strategy that can be effectively combined with advanced analytical tools, including Nuclear Magnetic Resonance spectroscopy and Liquid Chromatography-Mass Spectrometry, to enhance the characterization and detection capabilities. This marks the first instance of a simple method designed to detect 2'-5'-adenosine linked nucleic acids. The new method is characterized by its time-saving nature, simplicity, and relative accuracy compared to previous methods.

Targeted i6A-RNA degradation through sequential Fluorination-Azidation and Click reaction with imidazole-based probes.

We report the use of trimethylsilyl azide and Selectfluor to implement a standard protocol targeted at the prenylated nucleic acid known as i6A-RNA. After optimizing the conditions, we applied this method to regulate a wide range of i6A-RNA species using synthetic imidazole-based probes (I-IV). We observed that prenylated nucleic acid plays a crucial role in the cell hemostasis in A549 cell lines.

Multifunctional self-assembled adsorption microspheres based on waste bamboo shoot shells for multi-pollutant water purification.

In this study, multilayer self-assembled multifunctional bamboo shoot shell biochar microspheres (BSSBM) were prepared, in which bamboo shoot shell biochar was used as the carrier, titanium dioxide as the intermediate medium, and chitosan as the adhesion layer. The adsorption behavior of BSSBM on heavy metals Ag(I) and Pd(II), antibiotics, and dye wastewater was systematically analyzed. BSSBM shows a wide range of adsorption capacity. BSSBM is a promising candidate for the purification of real polluted water, not only for metal ions, but also for Tetracycline (TC) and Methylene Blue (MB). The maximum adsorption amounts of BSSBM on Pd(II), Ag(I), TC and MB were 417.3 mg/g, 222.5 mg/g, 97.2 mg/g and 42.9 mg/g, respectively.The adsorption of BSSBM on Pd(II), MB and TC conformed to the quasi-first kinetic model, and the adsorption on Ag(I) conformed to the quasi-second kinetic model. BSSBM showed remarkable selective adsorption capacity for Ag(I) and Pd(II) in a multi-ion coexistence system. BSSBM not only realized the high value-added utilization of waste, but also had the advantages of low cost, renewable and selective adsorption. BSSBM demonstrated its potential as a new generation of multifunctional adsorbent, contributing to the recovery of rare/precious metals and the treatment of multi-polluted water.

Sequential Azidation/Azolation of Prenylated Derivatives and a Click Reaction Enable Selective Labeling and Degradation of RAS Protein.

We propose the introduction of the azido and azo-functionalities into prenylated derivatives under mild conditions in a selective and efficient way. Upon protocol establishment and substrate scope determination, we apply this method to prenylated protein (citronellol-BSA) labeling, chemical pulldown, and enrichment. Eventually, we achieve the degradation of RAS on MCF-7 and HeLa cell lines by employing the well-designed probe von Hippel-Lindau derivatives C4 through the sequential azidation/azolation and click-reaction (SACR) pathway targeting the prenyl functionality attached to the Caax motif of the tested RAS protein. This method displays great potential in regulation of prenylated molecules.

Controlled synthesis of monodisperse gold nanorods with a small diameter of around 10 nm and largest plasmon wavelength of 1200 nm.

Gold nanorods have been widely used in various fields due to their tunable anisotropic localized surface plasmon resonance (SPR) property. The facile preparation of gold nanorods with a tunable SPR wavelength extending to a near-infrared window, and at the same time, a relatively small particle size for facilitating applications especially in the biomedical field is of great value yet highly challenging. In this work, a new reducing agent, 1,6-dihydroxynaphthalene, is proposed for the synthesis of gold nanorods. The results indicate that gold nanorods with good monodispersity, high shape yield, maximum SPR wavelength of 1200 nm, and especially small diameter of around 10 nm can be acquired simultaneously. In terms of spectral and size controls, by respectively varying the experimental parameters including the amount of silver ions, reducing agents, and gold seeds not only can a good linear correlation be acquired corresponding to a SPR wavelength ranging from around 600 nm to 1200 nm, but a regular change in the particle diameter from 10.5 nm to 7.5 nm could also be observed. The structural and morphological evolutions of the particle for each changed parameter were carefully studied, and insights were gained into the growth mechanism based on the detailed analysis of particle evolution at a specific stage of the growth process.

Comparison of the diagnostic efficiency between the O-RADS US risk stratification system and doctors' subjective judgment.

BACKGROUND: This study aimed to compare the diagnostic efficiency of Ovarian-Adnexal Reporting and Data System (O-RADS) and doctors' subjective judgment in diagnosing the malignancy risk of adnexal masses. METHODS: This was an analysis of 616 adnexal masses between 2017 and 2020. The clinical findings, preoperative ultrasound images, and pathological diagnosis were recorded. Each adnexal mass was evaluated by doctors' subjective judgment and O-RADS by two senior doctors and two junior doctors. A mass with an O-RADS grade of 1 to 3 was a benign tumor, and a mass with an O-RADS grade of 4-5 was a malignant tumor. All outcomes were compared with the pathological diagnosis. RESULTS: Of the 616 adnexal masses, 469 (76.1%) were benign, and 147 (23.9%) were malignant. There was no difference between the area under the curve of O-RADS and the subjective judgment for junior doctors (0.83 (95% CI: 0.79-0.87) vs. 0.79 (95% CI: 0.76-0.83), p = 0.0888). The areas under the curve of O-RADS and subjective judgment were equal for senior doctors (0.86 (95% CI: 0.83-0.89) vs. 0.86 (95% CI: 0.83-0.90), p = 0.8904). O-RADS had much higher sensitivity than the subjective judgment in detecting malignant tumors for junior doctors (84.4% vs. 70.1%) and senior doctors (91.2% vs. 81.0%). In the subgroup analysis for detecting the main benign lesions of the mature cystic teratoma and ovarian endometriosic cyst, the junior doctors' diagnostic accuracy was obviously worse than the senior doctors' on using O-RADS. CONCLUSIONS: O-RADS had excellent performance in predicting malignant adnexal masses. It could compensate for the lack of experience of junior doctors to a certain extent. Better performance in discriminating various benign lesions should be expected with some complement.

Assessing Obstetric Anal Sphincter Injuries: A Comparison of Exoanal and Endoanal Ultrasound.

OBJECTIVES: To explore the differences in assessing obstetric anal sphincter injuries (OASI) between transperineal ultrasound (TPUS) and endoanal ultrasound (EAUS) and test relationships between ultrasound findings and anal incontinence (AI) symptoms. METHODS: A group of 196 women with a history of vaginal delivery was recruited. OASI was detected in a set of 5 slices by EAUS and 8 slices by TPUS. OASI grading was performed on TPUS rules and EAUS rules. A "significant sphincter defect" was diagnosed by TPUS and EAUS using "2/3 rules." Symptoms of AI were determined using the St Mark's Incontinence Score (SMIS). Ultrasound findings were compared between the two methods and correlated with symptoms. RESULTS: Of 196 women, 29 (14.8%) suffered from AI with a mean SMIS of 12.1 ± 4.5, and 70 (35.7%) women with a mean age of 57 years had suspected OASI on imaging. Twenty-one (10.7%) "significant defects" were diagnosed by TPUS and 24 (12.2%) by EAUS. OASI Grades on TPUS had good agreement with EAUS rules (k = 0.70, P < .001). Logistic regression analysis showed that OASI Grade on imaging and "significant sphincter defects" seen on both forms of imaging were associated with AI symptoms. The odds ratio was 46 and 38 for "significant defects" on TPUS and EAUS, and 14 and 7 for OASI 3b+ on TPUS and EAUS in predicting AI, respectively. CONCLUSIONS: "Significant defects" diagnosed by EAUS or TPUS and OASI Grade 3b+ predict AI symptoms. The diagnostic performance of endoanal and exoanal ultrasound (EAUS and TPUS) appear to be very similar.

CD38+ B cells affect immunotherapy for allergic rhinitis.

BACKGROUND: Allergen-specific immunotherapy (AIT) is the mainstay in the treatment of allergic diseases, but the therapeutic effects of AIT need to be improved. CD38+ B cells are an immune cell fraction involved in the pathogenesis of allergic diseases as well as in immune regulation. OBJECTIVE: We sought to elucidate the role of antigen-specific CD38+ B cells in AIT. METHODS: An analysis was carried out on AIT results of 48 patients with perennial allergic rhinitis (AR), among which peripheral blood immune cells were analyzed by flow cytometry; serum cytokine levels were determined by ELISA. An AR murine model was developed to test the role of CD38+ B cells in AIT. RESULTS: A fraction of antigen-specific CD38+ B cell was detected in AR patients. CD38+ B-cell frequency was negatively correlated with the therapeutic effects of AIT. A negative correlation was detected between the CD38+ B-cell frequency and regulatory T-cell frequency in AR patients treated with AIT. Exposure to specific antigens induced CD38+ B cells to produce IL-6, that converted Treg cells to TH17 cells. Coadministration of anti-CD38 antibody significantly promoted the therapeutic effects of AIT. CONCLUSIONS: Antigen-specific CD38+ B cells compromise AIT effects by producing IL-6 to convert regulatory T cells to TH17 cells. Inhibition of CD38+ B cells promotes the effects of AIT.

Propionic acid regulates immune tolerant properties in B Cells.

Interleukin 10 (IL-10)-producing B cells (B10 cells) are a canonical cell fraction for regulating other activities of immune cells. Posttranscriptional modification of IL-10 in B10 cells is not yet fully understood. Short-chain fatty acids play an important role to regulate the functions of immune cells. This study aims to clarify the role of propionic acid (PA), a short-chain fatty acid, in regulating the expression of IL-10 in B10 cells. Blood samples were collected from patients with food allergy (FA) and healthy subjects. Serum and cellular components were prepared with the samples, and analysed by enzyme-linked immunosorbent assay and flow cytometry, respectively. The results showed that serum PA levels were lower in FA patients. PA concentrations were negatively correlated with serum cytokine Th2 concentrations, specific IgE concentrations in serum and skin prick test results. The peripheral frequency of B10 cells and the production of IL-10 in B cells were also associated with serum PA concentrations. Activation of B cells by CpG induced the production of IL-10 and tristetretrprolin (TTP), in which TTP caused the spontaneous decay of IL-10 mRNA. PA was necessary to stabilize the IL-10 mRNA in B cells by inducing the production of granzyme B, which resulted in the degradation of the IL-10 mRNA. Administration of PA attenuated FA response in mice by maintaining homeostasis of B10 cells. In conclusion, PA is needed to stabilize the expression of IL-10 in B10 cells. PA administration can mitigate experimental FA by maintaining B10 cell functions.

Nickel-Catalyzed 1,1-Dihydrophosphinylation of Nitriles with Phosphine Oxides.

Treatment of phosphine oxides with nitriles usually furnishes 1,2-dihydrophosphinylation products. Herein, we developed a nickel-catalyzed 1,1-dihydrophosphinylation of nitriles with phosphine oxides to access primary amines. This reaction proceeded smoothly under very mild conditions. A series of nitriles and phosphine oxides were compatible with this conversion, and the desired products were obtained in moderate to good yields.

Ultrasound Findings and O-RADS Malignancy Risk Stratification of Ovarian Collision Tumors.

OBJECTIVES: To describe the ultrasonographic signs of ovarian collision tumors and evaluate the malignancy risk using the O-RADS system. METHODS: This was a retrospective analysis of 25 ovarian collision tumors from 8739 patients between May 2010 and January 2020. All clinical characteristics, ultrasound images, and histological findings were collected and analyzed. Using the O-RADS lexicon descriptors, the O-RADS score was determined by two senior doctors. Lesions with O-RADS scores of 1 to 3 were classified as benign tumors, and lesions with O-RADS scores of 4 to 5 were classified as malignant tumors. RESULTS: The mean age of the 25 patients was 30.4 years. Histological findings showed that all tumors were a combination of mature cystic teratomas and cystadenomas/cystadenocarcinomas. There were 11 benign tumors of O-RADS 2 or 3, including 3 uniocular cysts, 3 dermoid cysts, and 5 dermoid cysts with an anechoic fluid cavity. There were 12 benign tumors of O-RADS 4, including 3 uniocular-multilocular cysts with solid components, 2 multilocular cysts with irregular walls, and 8 multilocular cysts with dermoid sacs. One borderline tumor of O-RADS 4 was a multilocular cyst with irregular septation. One malignant tumor of O-RADS 5 was a multilocular cystic tumor with solid components and ascites. CONCLUSIONS: The most common ovarian collision tumor was the coexistence of a mature cystic teratoma and a cystadenoma. The O-RADS system was able to sensitively detect malignant tumors in this series. A typical dermoid cyst with an anechoic cavity or a multilocular cyst with a dermoid sac may signify a benign collision tumor.

The Roles of Relative Linguistic Proficiency and Modality Switching in Language Switch Cost: Evidence from Chinese Visual Unimodal and Bimodal Bilinguals.

The current study investigated the mechanism of language switching in unbalanced visual unimodal bilinguals as well as balanced and unbalanced bimodal bilinguals during a picture naming task. All three groups exhibited significant switch costs across two languages, with symmetrical switch cost in balanced bimodal bilinguals and asymmetrical switch cost in unbalanced unimodal bilinguals and bimodal bilinguals. Moreover, the relative proficiency of the two languages but not their absolute proficiency had an effect on language switch cost. For the bimodal bilinguals the language switch cost also arose from modality switching. These findings suggest that the language switch cost might originate from multiple sources from both outside (e.g., modality switching) and inside (e.g., the relative proficiency of the two languages) the linguistic lexicon.

Association between long-term sedentary behavior and depressive symptoms in U.S. adults.

The study aimed to investigate the association between long-term sedentary behavior (LTSB) and depressive symptoms within a representative sample of the U.S. adult population. Data from NHANES 2017-2018 were used, encompassing information on demographics, depressive symptoms, physical activity (PA), and LTSB. Depressive symptoms were identified using the Patient Health Questionnaire (PHQ-9), with "depressive symptoms" defined as a PHQ-9 score of ≥ 5, and "moderate to severe depressive symptoms (MSDS)" defined as a PHQ-9 score of ≥ 10. PA and LTSB were assessed through the Global Physical Activity Questionnaire, where LTSB was interpreted as sedentary time ≥ 600 min. Restricted Cubic Spline (RCS) curves were utilized to observe potential nonlinear relationships. Binary Logistic regressions were conducted to analyze the associations. A total of 4728 participants (mean age 51.00 ± 17.49 years, 2310 males and 2418 females) were included in the study. Among these individuals, 1194 (25.25%) displayed depressive symptoms, with 417 (8.82%) exhibiting MSDS. RCS curves displayed increased risk of depressive symptoms with prolonged sedentary duration. Logistic regression models indicated significant associations between LTSB and depressive symptoms (OR 1.398, 95% CI 1.098-1.780), and LTSB and MSDS (OR 1.567, 95% CI 1.125-2.183), after adjusting for covariates. These findings suggest that LTSB may act as a potential risk factor for both depressive symptoms and MSDS in the studied population.

Propolis Alleviates Acute Lung Injury Induced by Heat-Inactivated Methicillin-Resistant Staphylococcus aureus via Regulating Inflammatory Mediators, Gut Microbiota and Serum Metabolites.

Propolis has potential anti-inflammatory properties, but little is known about its efficacy against inflammatory reactions caused by drug-resistant bacteria, and the difference in efficacy between propolis and tree gum is also unclear. Here, an in vivo study was performed to study the effects of ethanol extract from poplar propolis (EEP) and poplar tree gum (EEG) against heat-inactivated methicillin-resistant Staphylococcus aureus (MRSA)-induced acute lung injury (ALI) in mice. Pre-treatment with EEP and EEG (100 mg/kg, p.o.) resulted in significant protective effects on ALI in mice, and EEP exerted stronger activity to alleviate lung tissue lesions and ALI scores compared with that of EEG. Furthermore, EEP significantly suppressed the levels of pro-inflammatory mediators in the lung, including TNF-α, IL-1ß, IL-6, and IFN-γ. Gut microbiota analysis revealed that both EEP and EEG could modulate the composition of the gut microbiota, enhance the abundance of beneficial microbiota and reduce the harmful ones, and partly restore the levels of short-chain fatty acids. EEP could modulate more serum metabolites and showed a more robust correlation between serum metabolites and gut microbiota. Overall, these results support the anti-inflammatory effects of propolis in the treatment of ALI, and the necessity of the quality control of propolis.

A spherical adsorbent produced from a bagasse biochar chitosan assembly for selective adsorption of platinum-group metals from wastewater.

This study addresses the challenge of platinum-group metal scarcity by exploring the adsorption of these metals from industrial wastewater. An inexpensive adsorbent with selective platinum-group metal adsorption capacity, named chitosan/citric acid@diatomaceous earth-sugarcane bagasse (CTS/CA@DE-SBS), was newly synthesized. The material features a double coating of chitosan and diatomite on bagasse biochar, and it exhibits an excellent adsorption performance for platinum-group metals due to the synergistic effects of the biochar and chitosan-diatomaceous earth intercross-linked coatings. CTS/CA@DE-SBS achieved an 81 % adsorption efficiency and a static saturated adsorption capacity of 217 mg/g for Pt (IV) in water. Notably, the material exhibited selective adsorption properties for platinum-group metals dissolved in diverse aqueous solutions. The potential for the secondary recovery of platinum-group metals in complex aqueous bodies further underscores the significance of this adsorbent. In conclusion, this research introduces a promising solution for platinum-group metal shortages, offering a cost-effective and selective adsorbent with potential applications in the secondary recovery of these metals from industrial wastewater.

Degradation of Rat Sarcoma Proteins Targeting the Post-Translational Prenyl Modifications via Cascade Azidation/Fluorination and Click Reaction.

Protein degradation is emerging as a powerful strategy to modulate protein functions and alter cellular signaling pathways. Proteolysis-targeting chimeras (PROTACs) have been used to degrade a range of "undruggable" proteins in cells. Here, we present a type of chemically catalyzed PROTAC to induce rat sarcoma (RAS) degradation based on the chemistry of post-translational prenyl modification. Trimethylsilyl azide and Selectfluor were used to chemically tag the prenyl modification on Caax motif of RAS protein, and a sequential click reaction was applied using the propargyl pomalidomide probe to degrade the prenylated RAS in several cells. Thus, this approach was successfully applied to degrade RAS in multiple cancer cell lines including HeLa, HEK 293T, A549, MCF-7, and HT-29. This novel approach targeting RAS's post-translational prenyl modification to induce RAS degradation by employing the sequential azidation/fluorination and click reaction has been demonstrated efficiently and highly selectively, expanding PROTAC toolsets in the study of disease-relevant protein targets.

Integration with Transcriptomic and Metabolomic Analyses Reveals the In Vitro Cytotoxic Mechanisms of Chinese Poplar Propolis by Triggering the Glucose Metabolism in Human Hepatocellular Carcinoma Cells.

Natural products serve as a valuable reservoir of anticancer agents. Chinese poplar propolis (CP) has exhibited remarkable antitumor activities, yet its precise mechanisms of action remain elusive. This study aims to elucidate the in vitro cytotoxic mechanisms of CP in human hepatocellular carcinoma cells (HepG2) through comprehensive transcriptomic and metabolomic analyses. Our evidence suggested that CP possesses a great potential to inhibit the proliferation of HepG2 cells by targeting the glucose metabolism. Notably, CP exhibited a dose- and time-dependent reduction in the viability of HepG2 cells. Transcriptome sequencing unveiled significant alterations in the cellular metabolism, particularly within glucose metabolism pathways. CP effectively restrained glucose consumption and lactic acid production. Moreover, the CP treatment led to a substantial decrease in the mRNA expression levels of key glucose transporters (GLUT1 and GLUT3) and glycolytic enzymes (LDHA, HK2, PKM2, and PFK). Correspondingly, CP suppressed some key protein levels. Cellular metabolomic analysis demonstrated a marked reduction in intermediary products of glucose metabolism, specifically fructose 1,6-bisphosphate and acetyl-CoA, following CP administration. Finally, key compounds in CP were screened, and apigenin, pinobanksin, pinocembrin, and galangin were identified as potential active agents against glycolysis. It indicates that the effectiveness of propolis in inhibiting liver cancer is the result of the combined action of several components. These findings underscore the potential therapeutic value of propolis in the treatment of liver cancer by targeting glycolytic pathways.