Zhiheng Lin

1. The development of tumors is a multifactorial process involving genetic predisposition, environmental exposure and the dynamic interplay of immune dysregulation over time. Mounting evidence sugg...

Decidualization deficiency is a hallmark pathology of unexplained recurrent spontaneous abortion (URSA), but the undefined molecular drivers hinder the development of effective therapies. Hyperoside, a bioactive flavonoid from Hypericum perforatum, exhibits therapeutic potential against URSA, yet its underlying mechanism of action remains unknown. In this study, we employed an integrated multi‐omics approach coupled with a multi‐dimensional validation framework that spanned URSA patient decidual tissues, in vivo mouse models, and in vitro telomerase‐immortalized human endometrial stromal cell (T‐hESC) decidualization system, to systematically investigate hyperoside's mechanism in URSA, with a focus on R‐loop‐driven endometrial stromal cell senescence. We found that hyperoside dose‐dependently reduced embryo resorption and rescued decidualization deficiency by preventing stromal cell senescence. Mechanistically, hyperoside effectively alleviated aberrant intracellular R‐loop accumulation, thereby suppressing excessive activation of the cyclic GMP‐AMP synthase‐stimulator of interferon genes (cGAS‐STING) pathway which contributes to the initiation of the cellular senescence program. Further target identification and validation experiments confirmed that DExH‐box helicase 9 (DHX9) was the functional molecular target of hyperoside, with the Thr419 residue serving as the critical binding site. Functional validation revealed that DHX9 knockdown or introduction of the T419A point mutation markedly attenuated the anti‐senescence and pro‐decidualization effects of hyperoside. In vivo experiments further confirmed that uterine‐specific knockdown of DHX9 reduced hyperoside's protective effects against R‐loop accumulation, cGAS‐STING pathway activation, and embryo loss. Collectively, these findings demonstrate that hyperoside alleviates stromal cell senescence and decidualization deficiency in URSA through DHX9‐dependent resolution of R‐loops and subsequent suppression of cGAS‐STING‐associated senescence signaling. More broadly, this work identifies R‐loop‐mediated genomic stress as a previously underappreciated contributor to URSA‐associated decidual dysfunction and provides a mechanistic basis for the protective effects of hyperoside through DHX9‐dependent R‐loop homeostasis.

RNA‐targeted therapy is reshaping molecular medicine by shifting the traditional “protein‐centric” view toward an “RNA‐regulatory network” paradigm. Beyond carrying genetic information, RNA plays essential roles in posttranscriptional regulation, signaling pathways, and epigenetic modulation. Advances in high‐throughput sequencing, structural biology, and delivery technologies have accelerated the development of diverse RNA therapeutics, including antisense oligonucleotides (ASOs), small interfering RNA (siRNA), microRNA (miRNA) modulators, messenger RNA (mRNA) therapeutics, aptamers, short hairpin RNA, and CRISPR/Cas‐guided single‐guide RNAs. However, a concise comparison of these major RNA modalities and the translational barriers that limit their broader clinical application is still lacking. This review outlines the mechanisms and representative applications of these RNA‐based strategies in gene silencing, editing, protein replacement, immune activation, and targeted drug delivery. Special emphasis is placed on ASOs and siRNAs for neurological, metabolic, and infectious diseases, as well as mRNA therapeutics that are transforming vaccine development. Common challenges‐such as in vivo stability, delivery efficiency, and immune activation‐are also discussed. Finally, we highlight how chemical modification, nanotechnology, and artificial intelligence‐assisted design are enhancing the specificity, stability, and safety of RNA therapeutics, providing a framework for advancing next‐generation precision RNA medicine.

Background: Ovarian cancer (OC), as one of the gynecological malignancies, seriously threatens people’s life, health and safety with its high recurrence rate and high mortality rate. Methods: The GEO database served as the source of Single-cell RNA sequencing (scRNA-seq) data used in this study. We used scRNA-seq and spatial transcriptomics (ST), including CytoTRACE, Slingshot pseudotime analysis, CellChat, MultiNicheNet, pySCENIC, etc. From the perspective of time and space, the heterogeneity in OC was clarified, and key cell subgroups were obtained, as well as key transcription factors (TFs) and cell interaction pathways that control disease progression. Besides, this study further integrated clinical factors extracted from extensive RNA sequencing data in the TCGA database, and on this basis, constructed a prognostic model. Results: Through the combined analysis of scRNA-seq and ST techniques in this study, the key tumor subgroup, C4 SAA1 ⁺ tumor cells, was discovered. This subgroup mainly exists in untreated OC patients, and the SAA1 was closely related to the progression of OC. Furthermore, the key TF HOXA1 in this cell subpopulation was a risk factor in various cancers, including OC. We also revealed the correlations between different groups and the prognosis, immune infiltration, and drug sensitivity of patients by constructing risk-prognosis models, including high and low SAA1 ⁺ Tumor Cells Risk Score (STRS) groups. Conclusion: This study systematically mapped the single cell map of ovarian cancer. The identified key genes, cell subgroups, and the newly developed STRS prognostic model all showed significant clinical application value. These findings not only deepen the understanding of the pathogenesis of OC, but also provide a theoretical basis for optimizing diagnosis and treatment strategies, thus promoting the transformation from research to clinical practice to better meet the needs of patients.

Background: Glioblastoma (GBM), with its high recurrence and mortality rates, makes it the deadliest neurological malignancy. Oxidative phosphorylation is a highly active cellular pathway in GBM, and NFYB is a tumor-associated transcription factor. Both are related to mitochondrial function, but studies on their relationship with GBM at the single-cell level are still scarce.Methods: We re-analyzed the single-cell profiles of GBM from patients with different subtypes by single-cell transcriptomic analysis and further subdivided the large population of Glioma cells into different subpopulations, explored the interrelationships and active pathways among cell stages and clinical subtypes of the populations, and investigated the relationship between the transcription factor NFYB of the key subpopulations and GBM, searching for the prognostic genes of GBM related to NFYB, and verified by experiments.Results: Glioma cells and their C5 subpopulation had the highest percentage of G2M staging and rGBM, which we hypothesized might be related to the higher dividing and proliferating ability of both Glioma and C5 subpopulations. Oxidative phosphorylation pathway activity is elevated in both the Glioma and C5 subgroup, and NFYB is a key transcription factor for the C5 subgroup, suggesting its possible involvement in GBM proliferation and recurrence, and its close association with mitochondrial function. We also identified 13 prognostic genes associated with NFYB, of which MEM60 may cause GBM patients to have a poor prognosis by promoting GBM proliferation and drug resistance. Knockdown of the NFYB was found to contribute to the inhibition of proliferation, invasion, and migration of GBM cells.Conclusion: These findings help to elucidate the key mechanisms of mitochondrial function in GBM progression and recurrence, and to establish a new prognostic model and therapeutic target based on NFYB.

Objective: Endometrial cancer is one of the most common tumors of the female reproductive system, and the existing treatment options for advanced and metastatic endometrial cancer have certain limitations. The antitumor activity of luteolin has been gradually discovered. The purpose of this study was to predict the potential of luteolin in the treatment of endometrial cancer and to provide reference for future clinical drug use. Methods: The target gene database of luteolin and differential gene dataset of uterine corpus endometrial carcinoma (UCEC) have been constructed to obtain the differential genes (DR-DEGs) for luteolin and UCEC. The Gene Set Enrichment Analysis (GSEA), Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis are performed at the same time. Genes associated with prognosis in DR-DEGs were screened and validated using univariate and multivariate COX risk regression analysis so as to construct a prognostic model. Genes are divided into high-risk and low-risk groups according to risk scores for survival analysis and the predictive effect of the model is evaluated. The role of immune function in UCEC is investigated by immune infiltration and immune checkpoint analysis Finally, Transwell experiment was conducted to investigate the effect of luteolin on the migration ability of endometrial cancer cells, and the expression changes of MMP1, IL-17 and VEGF were detected by q-PCR. Results: Through the GO, KEGG and GSEA enrichment analysis, we have found a significant enrichment in "IL 17 signaling (IL-17) pathway", "oxidative stress response" and "HOMOLOGOUS_RECOMBINATION". Through multivariate COX risk regression analysis, four genes associated with the prognosis are harvested, including "PRSS1, MMP1, ERBB2 and NUF2" which belong to high-risk genes. Kaplan-Meier analysis shows that the survival rate in the high risk group is lower than that in the low risk group, and the receiver operating characteristic (ROC) curve reveals that the predictive effect of the model is good and stable (area under 1-year curve (AUC) 0.569, two-year AUC 0.628 and three-year AUC 0.653). Immune infiltration and immune checkpoint analysis suggest that "CD40", "T cells regulatory (Tregs)", "dendritic cells resting" and "dendritic cells activated" are correlated with survival and prognosis in UCEC patients. In in vitro experiments, we found that the migration ability of endometrial cancer cells was significantly reduced after luteolin treatment, and the expressions of MMP1, IL-17 and VEGF were all decreased. Conclusion: Through bioinformatic analysis, we found that luteolin could slow down the progression of UCEC by inhibiting the production of inflammatory mediators such as IL-17 and oxidative stress, and constructed genetic prognostic models associated with them: PRSS1, MMP1, ERBB2 and NUF2, respectively. In addition, we found that luteolin has an inhibitory effect on the migration of endometrial cancer cells and can reduce the expressions of MMP1, IL-17 and VEGF, thus easing the progression of endometrial cancer.

N-7 methylguanine (m7G) is one of the most common RNA base modifications in post-transcriptional regulation, which participates in multiple processes such as transcription, mRNA splicing and translation during the mRNA life cycle. However, its expression and prognostic value in uterine corpus endometrial carcinoma (UCEC) have not been systematically studied. In this paper, the data such as gene expression profiles, clinical data of UCEC patients, somatic mutations and copy number variants (CNVs) are obtained from the cancer genome atlas (TCGA) and UCSC Xena. By analyzing the expression differences of m7G-related mRNA in UCEC and plotting the correlation network maps, a risk score model composed of four m7G-related mRNAs (NSUN2, NUDT3, LARP1 and NCBP3) is constructed using least absolute shrinkage and selection operator (LASSO), univariate and multivariate Cox regression in order to identify prognosis and immune response. The correlation of clinical prognosis is analyzed between the m7G-related mRNA and UCEC via Kaplan–Meier method, receiver operating characteristic (ROC) curve, principal component analysis (PCA), t-SNE, decision curve analysis (DCA) curve and nomogram etc. It is concluded that the high risk is significantly correlated with (P < 0.001) the poorer overall survival (OS) in patients with UCEC. It is one of the independent risk factors affecting the OS. Differentially expressed genes are identified by R software in the high and low risk groups. The functional analysis and pathway enrichment analysis have been performed. Single sample gene set enrichment analysis (ssGSEA), immune checkpoints, m6A-related genes, tumor mutation burden (TMB), stem cell correlation, tumor immune dysfunction and rejection (TIDE) scores and drug sensitivity are also used to study the risk model. In addition, we have obtained 3 genotypes based on consensus clustering, which are significantly related to (P < 0.001) the OS and progression-free survival (PFS). The deconvolution algorithm (CIBERSORT) is applied to calculate the proportion of 22 tumor infiltrating immune cells (TIC) in UCEC patients and the estimation algorithm (ESTIMATE) is applied to work out the number of immune and matrix components. In summary, m7G-related mRNA may become a potential biomarker for UCEC prognosis, which may promote UCEC occurrence and development by regulating cell cycles and immune cell infiltration. It is expected to become a potential therapeutic target of UECE.

Necroptosis, a programmed form of necrotic cell death, plays critical regulatory roles in the progression and metastatic spread of cancers such as cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC). However, there are few articles systematically analyzing the necroptosis-related long non-coding RNAs (NRlncRNAs) correlated with CESC patients. Both RNA-sequencing and clinical data of CESC patients are downloaded from TCGA database in this study. Pearson correlation analysis, least absolute shrinkage, operator algorithm selection and Cox regression model are employed to screen and create a risk score model of eleven-NRlncRNAs (MIR100HG, LINC00996, SNHG30, LINC02688, HCG15, TUBA3FP, MIAT, DBH-AS1, ERICH6-AS1SCAT1, LINC01702) prognostic. Thereafter, a series of tests are carried out in sequence to evaluate the model for independent prognostic value. Gene set enrichment analytic paper, Gene Ontology analysis, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analytic paper make it clear that immune-related signaling pathways are very rich in the high-risk subgroup. Additionally, the prognostic risk score model is correlated to immune cell infiltration, potential immune checkpoint, immune function, immune micro-environmental and m6A-related gene. Mutation frequency in mutated genes and survival probability trend are higher in the low-risk subgroup in most of test cases when compared to the high-risk subgroup. This study constructs a renewed prognostic model of eleven-NRlncRNAs, which may make some contribution to accurately predicting the prognosis and the immune response from CESC patients, and improve the recognition of CESC patients and optimize customized treatment regimens to some extent.

Background: By using network pharmacology and molecular docking technology, we have explored the mechanism of action of Sanqi in the treatment of endometriosis (EMS), in order to provide reference for clinical studies of Chinese medicine treatment of Ems and Chinese medicine pharmacology. Methods: There are 123 intersecting targets between the active ingredients of Sanqi and disease targets. In the Protein-Protein Interaction network, Jun proto-oncogene, AP-1 transcription factor subunit, tumor necrosis factor, interleukin 6, etc., are the core proteins. The top 20 genes ranked by degree have been analyzed according to the Kyoto Encyclopedia of Genes and Genomes pathway and Gene Ontology analysis, and 20 pathways have been identified. Results: On the Kyoto Encyclopedia of Genes and Genomes pathway, the most important part is the phosphatidylinositol 3’-kinase-Akt signaling pathway, and on the Gene Ontology pathway, it is the Heme binding. The top 3 targets docked to quercetin have a certain affinity when it is docked to their degree value. Among the chemical components of Sanqi, quercetin has the most targets, suggesting that it may play a major role in the treatment of EMS. Conclusion: The results of molecular docking provide further evidence of the potential role of Sanqi for EMS. Overall, our study provides a new direction for the treatment of EMS and provides the basis for Sanqi as a drug for the treatment of EMS.

Background: The role of antioxidant intervention in polycystic ovary syndrome (PCOS) patients has been increasingly investigated in recent years. In order to further clarify whether antioxidant therapy is beneficial for PCOS patients and the emphasis of its effects, this study provides a systematic review and meta-analysis of randomized controlled trials examining the effect of antioxidant intervention on PCOS. Methods: Enrolled study designs related to antioxidant interventions and PCOS, published from 1999 to 2020, were searched from EMBASE, PubMed, and Web of Science databases to sort out proven studies on antioxidant interventions and PCOS. Data were reported as weighted mean difference (WMD) or standard mean difference with associated confidence intervals of 95%. The analysis was conducted using Stata version 16.0. Results: Twenty-three studies were included in total. Antioxidant intervention had a positive impact on homeostasis model assessment of insulin resistance (WMD = –0.37, P = .011) and Triglycerides (WMD = –25.51, P < .001). And antioxidant intervention did not improve testosterone levels significantly (WMD = –0.20, P = .2611). Subgroup analysis showed that except for the D-chiro-inosito subgroup, no difference in body mass index was observed between the intervention group and the control group. Conclusions: This meta-analysis demonstrates the efficacy of antioxidant intervention in patients with PCOS, demonstrating that antioxidant intervention has a significant effect on insulin resistance and lipid metabolism improvement. However, antioxidant intervention therapy has no discernible impact on testosterone levels or body mass index. Omega-3 may be a more effective antioxidant intervention for PCOS. In addition, this meta-analysis provides important reference opinions and treatment recommendations for PCOS.