Colorectal cancer， a leading malignant gastrointestinal tumor globally in terms of incidence and mortality， has seen a consistent annual rise in newly diagnosed cases. While conventional therapies like radiotherapy， chemotherapy， and surgery are available， problems such as lack of early diagnosis， poor prognosis， and drug resistance remain significant burdens for patients. Given the complex and diverse pathogenesis of colorectal cancer， there is an urgent clinical need for safe， effective， reliable， and multi-targeted therapeutic strategies. The Hippo signaling pathway， closely linked to mechanisms like tumorigenesis， cancer cell invasion， migration， and drug resistance， extensively participates in the occurrence and development of colorectal cancer， so targeting the signaling pathway for cancer prevention and treatment has become a crucial research direction in recent years. Traditional Chinese medicine （TCM） offers multi-faceted， multi-pathway， and multi-target advantages and becomes an important therapy for colorectal cancer by enhancing patients' immunity， improving the life quality， and prolonging survival. Studies show that the active components of TCM， including flavonoids， terpenoids， phenols， alkaloids， quinones， lignans， and saponins， as well as TCM compounds such as modified Sijunzi decoction， Jiedu Sangen decoction， Jianpi Jiedu compound， and Quyu Jiedu decoction， exhibit significant targeting effects on the Hippo signaling pathway. These TCMs can exert an anti-colorectal cancer effect through various mechanisms， such as inducing cancer cell autophagy and apoptosis， inhibiting epithelial-mesenchymal transition， reversing drug resistance of the tumor， and blocking the cancer cell cycle. This paper reviewed and analyzed Chinese and international research on the action mechanisms of TCM in regulating the Hippo signaling pathway for the prevention and treatment of colorectal cancer with a comprehensive overview presentation， aiming to provide new references and ideas for the clinical application of TCM and the development of new pharmacological agents in the prevention and treatment of colorectal cancer.

Colorectal cancer， a leading malignant gastrointestinal tumor globally in terms of incidence and mortality， has seen a consistent annual rise in newly diagnosed cases. While conventional therapies like radiotherapy， chemotherapy， and surgery are available， problems such as lack of early diagnosis， poor prognosis， and drug resistance remain significant burdens for patients. Given the complex and diverse pathogenesis of colorectal cancer， there is an urgent clinical need for safe， effective， reliable， and multi-targeted therapeutic strategies. The Hippo signaling pathway， closely linked to mechanisms like tumorigenesis， cancer cell invasion， migration， and drug resistance， extensively participates in the occurrence and development of colorectal cancer， so targeting the signaling pathway for cancer prevention and treatment has become a crucial research direction in recent years. Traditional Chinese medicine （TCM） offers multi-faceted， multi-pathway， and multi-target advantages and becomes an important therapy for colorectal cancer by enhancing patients' immunity， improving the life quality， and prolonging survival. Studies show that the active components of TCM， including flavonoids， terpenoids， phenols， alkaloids， quinones， lignans， and saponins， as well as TCM compounds such as modified Sijunzi decoction， Jiedu Sangen decoction， Jianpi Jiedu compound， and Quyu Jiedu decoction， exhibit significant targeting effects on the Hippo signaling pathway. These TCMs can exert an anti-colorectal cancer effect through various mechanisms， such as inducing cancer cell autophagy and apoptosis， inhibiting epithelial-mesenchymal transition， reversing drug resistance of the tumor， and blocking the cancer cell cycle. This paper reviewed and analyzed Chinese and international research on the action mechanisms of TCM in regulating the Hippo signaling pathway for the prevention and treatment of colorectal cancer with a comprehensive overview presentation， aiming to provide new references and ideas for the clinical application of TCM and the development of new pharmacological agents in the prevention and treatment of colorectal cancer.

Colorectal cancer is a common malignant tumor of the digestive system. In recent years， its incidence rate and mortality are increasing year by year. Due to the complex pathogenesis and poor prognosis of patients， colorectal cancer poses a serious threat to human physical and mental health. Currently， although Western medicine treatment methods can to some extent inhibit tumor growth and alleviate patient symptoms， postoperative recurrence， metastasis， multiple adverse reactions， and susceptibility to drug resistance are prominent issues， resulting in unsatisfactory overall treatment outcomes. Therefore， exploring more efficient and safe treatment methods has become an urgent task. The adenosine monophosphate-activated protein kinase （AMPK） signaling pathway plays a regulatory role in the growth， differentiation， apoptosis， and autophagy of colorectal cancer cells， and is widely involved in the occurrence and development of colorectal cancer. It is considered an important target for colorectal cancer treatment. Traditional Chinese medicine has unique advantages in the treatment of colorectal cancer， as it can exert its effects through multiple mechanisms and pathways. It can prevent postoperative recurrence and metastasis， reduce adverse reactions to radiotherapy and chemotherapy， and improve patients' quality of life. It has become a key means of treating colorectal cancer. Research has shown that active ingredients in traditional Chinese medicine such as flavonoids， polyphenols， terpenes， and esters， as well as traditional Chinese medicine compounds such as Qingjie Fuzheng Granules and some traditional Chinese medicine extracts， have significant regulatory effects on AMPK and its interaction signaling pathways. They exert their anti-colorectal cancer effects by inducing autophagy and apoptosis in colorectal cancer cells， promoting ferroptosis， inhibiting epithelial-mesenchymal transition， reversing drug resistance， and arresting the cell cycle. This article reviewed and summarized the relevant research on traditional Chinese medicine in the treatment of colorectal cancer in recent years， with a focus on the mechanism of traditional Chinese medicine in regulating the AMPK signaling pathway for the treatment of colorectal cancer. It is expected to provide ideas and references for the development of new drugs for clinical anti-colorectal cancer treatment.

Liver cancer， as a malignant tumor with high incidence and mortality worldwide， has shown a year-by-year increase in both incidence and mortality in China. Currently， Western medicine has achieved certain efficacy in the clinical management of liver cancer in terms of inhibiting tumor growth， alleviating patient symptoms， and delaying disease progression. However， its limitations cannot be ignored. Significant side effects， increasingly severe drug resistance， and limited improvement in patient survival collectively make the overall therapeutic outcomes fall short of ideal expectations. Therefore， exploring more effective and safer treatment strategies has become a critical issue that urgently needs to be addressed in the field of oncology. The adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） signaling pathway plays a central regulatory role in the physiological processes of liver cancer cells， including proliferation， differentiation， apoptosis， and autophagy， and is considered a key molecular target for anti-liver cancer therapy. Traditional Chinese medicine （TCM）， characterized by its multi-pathway and multi-mechanism synergistic effects， has become an important component of the current comprehensive treatment system for liver cancer. Among these， TCM monomeric compounds， owing to their well-defined chemical structures and precisely analyzable pharmacological mechanisms， represent an important breakthrough in anti-liver cancer drug development. Studies have shown that various TCM monomeric compounds， including flavonoids， alkaloids， polyphenols， saponins， terpenoids， and quinones， can regulate the AMPK/mTOR signaling pathway and its upstream and downstream protein expression. Through these mechanisms， they induce autophagy and apoptosis in liver cancer cells， inhibit aerobic glycolysis， reverse drug resistance， promote ferroptosis， block epithelial-mesenchymal transition， and inhibit angiogenesis， thereby effectively suppressing the growth and metastasis of liver cancer cells. Based on this， this article systematically reviews recent studies on TCM monomeric compounds in the treatment of liver cancer， and further analyzes in depth their mechanisms in regulating the AMPK/mTOR signaling pathway， so as to provide ideas and references for the development of new anti-liver cancer drugs.

Liver cancer， as a malignant digestive tract tumor with high incidence and mortality rates globally， has witnessed continuous increases in both incidence and mortality in recent years. Although modern medical treatment methods have achieved certain results in disease control， the treatment of liver cancer remains challenging due to the low early diagnosis rate， obvious toxic and side effects， and widespread drug resistance. Against this backdrop， exploring more efficient and safer treatment strategies is of utmost urgency. The Hippo cascade signaling pathway plays a crucial regulatory role in processes such as the proliferation， differentiation， apoptosis， and invasion of liver cancer cells. It is extensively involved in the occurrence and development of liver cancer， serving as an important target for anti-liver cancer treatment research. Traditional Chinese medicine （TCM） can exert anti-liver cancer effects through multiple approaches， mechanisms， and pathways. It has shown remarkable efficacy in preventing postoperative recurrence and metastasis of liver cancer， reducing adverse reactions of radiotherapy and chemotherapy， and improving the quality of life， thus becoming a key component of comprehensive liver cancer treatment. Studies have confirmed that the TCM active components including flavonoids， alkaloids， terpenoids， polyphenols， saponins， and quinones， as well as the TCM compound prescriptions such as compound Banmao capsules， Erzhu Jiedu decoction， Pianzaihuang， and Zhenggan prescription， can effectively regulate the Hippo signaling pathway. They exert significant anti-liver cancer effects through multiple mechanisms， such as inducing autophagy and apoptosis of liver cancer cells， improving macrophage polarization， blocking epithelial-mesenchymal transition， inhibiting glycolytic metabolism， promoting immune cell activation， reversing tumor drug resistance， and arresting the cell cycle. This study used "liver cancer" "Hippo" "signaling pathway" and "traditional Chinese medicine" as the main keywords to search multiple literature databases， including PubMed， CNKI， and VIP. By discussing and analyzing the mechanisms of TCM in regulating the Hippo signaling pathway for the treatment of liver cancer in the past five years， this paper aims to provide ideas and references for the research and development of new anti-liver cancer drugs in clinical practice.

ObjectiveTransfer RNA-derived fragments (tRFs) are a recently characterized and rapidly expanding class of small non-coding RNAs, typically ranging from 13 to 50 nucleotides in length. They are derived from mature or precursor tRNA molecules through specific cleavage events and have been implicated in a wide range of cellular processes. Increasing evidence indicates that tRFs play important regulatory roles in gene expression, primarily by interacting with target messenger RNAs (mRNAs) to induce transcript degradation, in a manner partially analogous to microRNAs (miRNAs). However, despite their emerging biological relevance and potential roles in disease mechanisms, there remains a significant lack of computational tools capable of systematically predicting the interaction landscape between tRFs and their target mRNAs. Existing databases often rely on limited interaction features and lack the flexibility to accommodate novel or user-defined tRF sequences. The primary goal of this study was to develop a machine learning based prediction algorithm that enables high-throughput, accurate identification of tRF:mRNA binding events, thereby facilitating the functional analysis of tRF regulatory networks. MethodsWe began by assembling a manually curated dataset of 38 687 experimentally verified tRF:mRNA interaction pairs and extracting seven biologically informed features for each pair: (1) AU content of the binding site, (2) site pairing status, (3) binding region location, (4) number of binding sites per mRNA, (5) length of the longest consecutive complementary stretch, (6) total binding region length, and (7) seed sequence complementarity. Using this dataset and feature set, we trained 4 distinct machine learning classifiers—logistic regression, random forest, decision tree, and a multilayer perceptron (MLP)—to compare their ability to discriminate true interactions from non-interactions. Each model’s performance was evaluated using overall accuracy, receiver operating characteristic (ROC) curves, and the corresponding area under the ROC curve (AUC). The MLP consistently achieved the highest AUC among the four, and was therefore selected as the backbone of our prediction framework, which we named tRF Prospect. For biological validation, we retrieved 3 high-throughput RNA-seq datasets from the gene expression omnibus (GEO) in which individual tRFs were overexpressed: AS-tDR-007333 (GSE184690), tRF-3004b (GSE197091), and tRF-20-S998LO9D (GSE208381). Differential expression analysis of each dataset identified genes downregulated upon tRF overexpression, which we designated as putative targets. We then compared the predictions generated by tRF Prospect against those from three established tools—tRFTar, tRForest, and tRFTarget—by quantifying the number of predicted targets for each tRF and assessing concordance with the experimentally derived gene sets. ResultsThe proposed algorithm achieved high predictive accuracy, with an AUC of 0.934. Functional validation was conducted using transcriptome-wide RNA-seq datasets from cells overexpressing specific tRFs, confirming the model’s ability to accurately predict biologically relevant downregulation of mRNA targets. When benchmarked against established tools such as tRFTar, tRForest, and tRFTarget, tRF Prospect consistently demonstrated superior performance, both in terms of predictive precision and sensitivity, as well as in identifying a higher number of true-positive interactions. Moreover, unlike static databases that are limited to precomputed results, tRF Prospect supports real-time prediction for any user-defined tRF sequence, enhancing its applicability in exploratory and hypothesis-driven research. ConclusionThis study introduces tRF Prospect as a powerful and flexible computational tool for investigating tRF:mRNA interactions. By leveraging the predictive strength of deep learning and incorporating a broad spectrum of interaction-relevant features, it addresses key limitations of existing platforms. Specifically, tRF Prospect: (1) expands the range of detectable tRF and target types; (2) improves prediction accuracy through multilayer perceptron model; and (3) allows for dynamic, user-driven analysis beyond database constraints. Although the current version emphasizes miRNA-like repression mechanisms and faces challenges in accurately capturing 5'UTR-associated binding events, it nonetheless provides a critical foundation for future studies aiming to unravel the complex roles of tRFs in gene regulation, cellular function, and disease pathogenesis.

ObjectiveTo explore the regulation of hypothalamic-pituitary-gonadal （HPG） axis by modified Erxian decoction in rats with perimenopausal syndrome （PMS） and to further analyze the expression of proteins related to the silent information regulator 1 （SIRT1）/hypothalamic kisspeptin （Kisspeptin）/gonadotropin-releasing hormone （GnRH） signaling pathway in the arcuate nucleus region （ARC） of the hypothalamus， so as to reveal the potential target of action and molecular biological mechanism of modified Erxian decoction for the treatment of perimenopausal syndrome. MethodsAn animal model was established via the incomplete castration method， with successful modeling confirmed by the exfoliated cervical cell smear method. The 48 rats were divided into six groups based on the randomization principle after successful modeling， including a sham operation group， a model group， an estradiol valerate group （0.09 mg∙kg^-1∙d^-1）， high-， medium-， and low-dose modified Erxian decoction groups （7.614， 3.807，1.903 5 g∙kg^-1∙d^-1）， with 8 rats in each group. The estradiol valerate group and the high-， medium- and low-dose modified Erxian decoction groups were continuously administered by gavage for 28 days， and the indicators were detected 24 hours after the last administration. Body weights and uterine indices were measured. The pathological changes of the uterus were observed by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was performed to measure the levels of estradiol （E₂）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and gonadotropin-releasing hormone （GnRH）. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to determine the expression levels of SIRT1， Kisspeptin， kisspeptin receptor （GPR54）， and GnRH in the ARC region of the hypothalamus and gonadotropin-releasing hormone receptor （GnRH-R） in pituitary. ResultsCompared with the sham operation group， rats in the model group had a significantly increased body weight （P0.01）， reduced wet weight and index of uterus （P0.01）， endometrial thinning or atrophy， glandular atrophy， and a decreasing number of glands. Additionally， serum levels of E₂ and the expression of SIRT1 in the ARC region of the hypothalamus significantly decreased （P0.01）. Serum levels of FSH， LH， and GnRH， the expression of Kisspeptin， GPR54， and GnRH in the ARC region of the hypothalamus， and GnRH-R in pituitary significantly increased （P0.01）. Compared with the model group， the estradiol valerate group and the high-， medium-dose modified Erxian decoction groups had significantly reduced body weight， serum levels of FSH， LH， and GnRH， and expression of Kisspeptin， GPR54， and GnRH in the ARC region of the hypothalamus and GnRH-R in pituitary （P0.05， P0.01） and significantly increased wet weight and index of uterus， serum level of E₂， and expression of SIRT1 in the ARC region of the hypothalamus （P0.05， P0.01）. In addition， they showed thickened endometrium， increased number of endometrial glands， and improved glandular atrophy. ConclusionModified Erxian decoction regulates the function of the HPG axis through multi-targets， and its mechanism of action may be related to the up-regulation of the expression of SIRT1 in the ARC region of the hypothalamus， the inhibition of the over-activation of the Kisspeptin/GnRH signaling pathway， the regulation of the expression of GnRH-R in the pituitary， the restoration of secretion balance of gonadotropins， and the elevation of the estrogen level. This study provides an experimental basis for the interpretation of the scientific connotation of modified Erxian decoction in the treatment of perimenopausal syndrome and a theoretical reference for the development of a novel therapeutic strategy based on the SIRT1/Kisspeptin/GnRH pathway.

Hepatocellular carcinoma （HCC） is one of the most common malignant tumors with high incidence and mortality rates worldwide， which brings a huge burden to the physical and mental health and socio-economic life of patients. The adenosine monophosphate-activated protein kinase （AMPK） signaling pathway serves as the regulatory center of cellular energy metabolism and is closely associated with the biological activities of HCC cells， including autophagy， apoptosis， and angiogenesis， and it has become a hot topic in current cancer research. Traditional Chinese medicine drugs are abundant in natural components such as flavonoids， alkaloids， and phenols and have the characteristics of multiple targets， pathways， components， and hierarchies. By targeting the AMPK signaling pathway， these components can be used alone or in combination with conventional antitumor therapies to exert an anti-tumor effect on HCC from various aspects. This article reviews and summarizes the extracts of traditional Chinese medicine that target the AMPK signaling pathway for the prevention and treatment of HCC， in order to provide a theoretical basis and a reference for the clinical application of traditional Chinese medicine in the treatment of HCC and the development of related drugs.

Objective:To investigate the effects of enolase 1(ENO1)on the proliferation,migration,and invasion of gastric cancer cells and its underlying molecular mechanisms.Methods:The expression levels of ENO1 in human gastric cancer cell lines(HGC27,MKN-45,N-87,MGC803,BGC-823)and human gastric mucosal epithelial cells(GES-1)were detected using WB assay.Gene editing tools such as CRISPR and overexpression system were used to construct ENO1 knockdown and knockdown-rescue cell lines.Both MKN-45 and BGC-823 cells were grouped into control(Ctrl)group,ENO1 knockdown(ENO1 KD)group,and ENO1 knockdown-rescue(ENO1 KD-OE)group.The effects of ENO1 knockdown or ENO1 knockdown-rescue on the proliferation,migration,invasion,and apoptosis of gastric cancer cells were evaluated using colony formation assay,EdU staining,scratch wound healing assay,Transwell chamber assay and flow cytometry.Additionally,a xenograft model was established in nude mice,and the effects of ENO1 on tumor growth were monitored using small animal in vivo imaging and tumor tissue block measurement.ENO1 was silenced in MKN-45 cells employing RNA interference technology,and the downstream target genes of ENO1 were identified using RNA co-immunoprecipitation sequencing(RIP-seq)and bioinformatics analysis.The molecular mechanisms by which ENO1 regulates the proliferation,migration and invasion of gastric cancer cells was also analyzed.Results:ENO1 was significantly upregulated in gastric cancer cell lines(P<0.01 or P<0.001).ENO1 knockdown significantly inhibited proliferation,migration,and invasion while promoting apoptosis in MKN-45 and BGC-823 cells(P<0.001,P<0.000 1).Rescue experiments showed that restoring ENO1 expression significantly enhanced cell proliferation,migration,invasion,and inhibited apoptosis(P<0.05,P<0.01,P<0.001,P<0.000 1).In vivo experiments demonstrated that ENO1 knockdown significantly inhibited tumor growth in nude mice(P<0.000 1).The differentially expressed genes interacting with ENO1 protein were primarily enriched in pathways related to RNA splicing.Additionally,ENO1 protein was found to interact with the PKM gene,and their expressions showed a positive correlation in gastric cancer tissues(r=0.886).Conclusion:ENO1 is highly expressed in gastric cancer cells.ENO1 interacts with precursor mRNA of PKM to influence its RNA splicing process,thereby regulating PKM2 expression and promoting gastric cancer progression.

Objective·To detect the expression level of Fas-associated protein with death domain(FADD)in head and neck squamous cell carcinoma(HNSCC)and to explore the molecular mechanisms by which FADD promotes the proliferation of HNSCC cells.Methods·The GEPIA 2 database was utilized to analyze the expression level of FADD in tumor tissues and to evaluate its association with prognosis.Immunohistochemistry staining(IHC)was performed on HNSCC tissues to investigate the changes in FADD expression levels in normal,dysplastic,and tumor tissues.Stable FADD-knockdown Fadu and HSC3 cell lines were constructed and validated using Western blotting and quantitative real-time PCR(qRT-PCR).The regulatory effect of FADD on the proliferation of HNSCC cells was explored using the LiveCyte live-cell tracking system,colony formation assay,and cell viability assay.Proteins interacting with FADD were identified by co-immunoprecipitation mass spectrometry(Co-IP/MS),and further mechanistic studies were conducted using CRISPR/Cas9 technology,LiveCyte live-cell tracking system,and Western blotting.Results·Analysis of the GEPIA2 database indicated that FADD was significantly overexpressed in head and neck cancer and was associated with poor prognosis.IHC staining showed that FADD expression levels progressively increased from normal to dysplastic to tumor tissues in HNSCC patients.Knockdown of FADD in HNSCC cells resulted in significantly reduced proliferation and colony formation compared to the control group.Co-IP/MS results showed that FADD interacted with the CUX1 protein,and FADD knockdown led to increased CUX1 expression.Moreover,CUX1 knockdown significantly promoted HNSCC cell proliferation and reversed the anti-proliferative phenotype caused by FADD knockdown.Conclusion·FADD plays a significant pro-carcinogenic role in HNSCC and is associated with poor prognosis.FADD can further regulate tumor cell proliferation by interacting with CUX1 and suppressing its expression level.