Yan Gao

Introduction: Microbiome multi-omics studies frequently identify microbial, metabolic, and host transcriptional associations, but their reproducibility and support for connected biological mechanisms remain uncertain. We evaluated these relationships in hepatocellular carcinoma (HCC) and colorectal cancer (CRC) using an eight-dimensional, non-compensatory evidence-gating framework. Methods: Nine public HCC datasets spanning microbiome, metabolome, and host transcriptome profiles were analyzed alongside CRC microbial and metabolomic data and separate host transcriptomic datasets. Analyses included cross-cohort replication, participant-matched taxon–metabolite associations, host transcriptional responses, sensitivity testing, and independent evidence scoring. Complete mechanistic eligibility required support across all eight evidence dimensions. Results: In HCC, 13 genera showed concordant directions and nominal significance across conventional leave-one-cohort-out analyses, but none met the adjusted external validation criterion. Six longitudinal plasma feature-window signals and 12 differential liver metabolites were identified without cross-specimen concordance under false discovery rate (FDR) correction. Three hepatic transcriptional programs replicated across independent bulk datasets, but none of the four HCC candidate families met all eight evidence requirements. In CRC, four bacterial species replicated in held-out disease-feature testing. Paired microbial and fecal metabolite profiles from 347 participants yielded 18 replicated taxon–metabolite associations, of which 10 met robustness criteria and eight retained support after joint-taxon adjustment. The Peptostreptococcus stomatis–glycocholate association replicated in held-out testing (adjusted residual r = 0.315, q = 0.00232) and remained significant after joint-taxon adjustment (β = 0.209, q = 0.0144) and unified-partition analysis with global Benjamini–Hochberg correction (held-out q = 0.0303). Reanalysis of glycocholic acid (GCA) treatment transcriptomic data identified increased SOX14, ZDHHC9, and CD274 expression, although the three-gene response score yielded an exact permutation P = 0.10. Independent assessments agreed on all 48 evidence ratings (quadratic-weighted κ = 1.000). Discussion: Evidence gating distinguished reproducible molecular associations from complete cross-layer mechanistic support. The P. stomatis–glycocholate association, accompanied by complementary GCA-responsive host transcription, represents a candidate for further investigation rather than an established microbial–metabolite–host mechanism.

Background: Carrier-free self-assembled nanodrugs use therapeutic or bioactive molecules as principal structural components. Their architectures have diversified rapidly, but how molecular design relates to resistance evidence and further development remains unclear. Objective: To map molecular architecture, drug-resistance evidence, and translation-relevant reporting in carrier-free anticancer nanodrugs. Methods: We conducted a PRISMA-ScR systematic scoping review and evidence map integrated with bibliometric analysis and structured appraisal of key resistance studies. PubMed, Europe PMC, Scopus, and Web of Science Core Collection (WoSCC) were searched in August 2026. WoSCC records (n = 863) characterized publication and intellectual structure, Scopus records (n = 840) assessed coverage, and 526 eligible articles formed the evidence map. Architecture was classified in 481 original studies, resistance evidence in 160 records, and 19 translation-relevant domains in 147 full-text studies. All 43 Tier 1/2 studies underwent full-report appraisal. Results: Covalent prodrug/conjugate assemblies (n = 187) and noncovalent multi-active co-assemblies (n = 182) predominated. Of 160 resistance-related records, 28 used established resistant models (Tier 1), whereas 15 provided mechanism-aligned sensitization evidence without resistant-model confirmation (Tier 2). Architecture was not clearly associated with Tier one versus Tier 2 classification in exploratory analysis (Cramér’s V = 0.394, Holm-adjusted P = 0.666). In the 147-study full-text subset, batch consistency and experimental scalability were each reported in three studies and long-term toxicity in four. Seven studies met the composite human-related evidence domain, but only one involved direct clinical administration (1/147, 0.7%). After excluding incomplete 2026 data, the latest-versus-earliest difference in reporting completeness was 1.40 domains (95% CI, −0.49–3.25), providing no clear evidence of sustained temporal improvement. Conclusion: Carrier-free anticancer nanodrugs have diversified rapidly, but evidence supporting resistance and further development has not matured in parallel. By separating resistance mechanism from evidentiary strength and distinguishing resistant-model validation from mechanism-aligned sensitization, this review establishes clearer boundaries for resistance claims and identifies manufacturing reproducibility, long-term safety, and direct clinical evaluation as major evidence gaps. The resulting framework provides a practical basis for aligning molecular design with claim-specific pharmacology and development-oriented evidence generation.

Context: Shengmai San (SMS) is a traditional Classic Recipes made with Panax ginseng, Ophiopogon, and Schisandra. The precise mechanism of action of SMS remains unclear, despite its noteworthy therapeutic advantages for type 2 diabetes mellitus (T2DM). Objective: The objective of this study was to confirm the mechanism of SMS in the treatment of T2DM. Materials and Methods: UPLC-MS/MS was employed to identify the active components in SMS. Using network pharmacology, the intervention pathways of SMS in T2DM rats were investigated. A high-sugar, high-fat diet and intrabitoneal injection of streptozotocin were used to create a T2DM rat model. The serum parameters of each group were assessed following the completion of the experiment. The mRNA and protein expression levels of genes related to the PI3K/AKT/GSK3B pathway were analyzed using qRT-PCR and Western blot. Results: There were 39 components found in SMS San by UPLC-Q-Orbitrap/MS. Network pharmacology and molecular docking study indicate that the PI3K/AKT/GSK3B pathway may be a part of SMS’s therapeutic mechanism for the treatment of T2DM. Rats’ serum levels of TC, TG, LDL-C, MDA, TNF-α, and IL-6 dramatically dropped after taking SMS, whereas SOD and HDL-C levels rose. The improvement of these parameters may be related to SMS activation of downstream target proteins, PI3K, and AKT. Conclusion: SMS can help cure T2DM mellitus by regulating the PI3K/AKT/GSK3B pathway, improving islet tissue injury, reducing oxidative stress, and easing lipid metabolism issues. This study not only revealed the unique therapeutic mechanism of SMS by regulating the PI3K/AKT/GSK3β signaling pathway, but also provided new theoretical support for multi-target and personalized treatment of diabetes. Its novelty lies in the first exploration of the mechanism of action of traditional Chinese medicine prescriptions through the lens of modern molecular biology, emphasizing the potential of natural medicines in diabetes treatment.

Lonicerae japonicae Flos (LJF) possesses a good anti-respiratory syncytial virus (RSV) effect. However, the material basis of LJF in treating RSV is still unclear. In this study, a sensitive and accurate quantitative method based on UHPLC-QQQ MS was established and validated for the simultaneous determination of the 15 ingredients from LJF in RSV-infected mice plasma. Multiple reaction monitoring was performed for quantification of the standards and of the internal standard in plasma. All the calibration curves show good linear regression within the linear range (r² > 0.9918). The method validation results, including specificity, linearity, accuracy, precision, extraction recovery, matrix effect, and stability of 15 ingredients, are all within the current acceptance criteria. This established method was successfully applied to the pharmacokinetic study of 15 compounds from LJF. Furthermore, the repair rate of lung index and the improvement rate of IFN-γ and IL-6 improved after administration of the LJF, indicating that LJF possessed a positive effect on the treatment of RSV infection. Finally, by combining Spearman and Grey relation analysis, isochlorogenic acid B, isochlorogenic acid C, secoxyloganin, chlorogenic acid, and loganic acid are speculated to be the main effective ingredients of LJF in treating RSV. This study lays the foundation for attempts to reveal the mechanisms of the anti-RSV effect of LJF.

Background: Transdermal delivery is very important in pharmaceutics. However, the barrier function of the stratum corneum hinders drugs absorption. How to improve transdermal delivery efficiency is a hot topic. The key advantages of physical technologies are their wide application for the delivery of previously nonappropriate transdermal drugs, such as proteins, peptides, and hydrophilic drugs. Based on the improved permeation of drugs delivered via multiple physical techniques, many more diseases may be treated, and transdermal vaccinations become possible. However, their wider application depends on the related convenient and portable devices. Combined products comprising medicine and devices represent future commercial directions of artificial intelligence and 3D printing. Methods: A comprehensive search about transdermal delivery assisted by physical techniques has been carried out on Web of Science, EMBASE database, PubMed, Wanfang Database, China National Knowledge Infrastructure, and Cochrane Library. The search identified and retrieved the study describing multiple physical technologies to promote transdermal penetration. Results: Physical technologies, including microneedles, lasers, iontophoresis, sonophoresis, electroporation, magnetophoresis, and microwaves, are summarized and compared. The characteristics, mechanism, advantages and disadvantages of physical techniques are clarified. The individual or combined applicable examples of physical techniques to improve transdermal delivery are summarized. Conclusion: This review will provide more useful guidance for efficient transdermal delivery. More therapeutic agents by transdermal routes become possible with the assistance of various physical techniques.