Chemical immunosuppressants have been widely used for the treatment of systemic lupus erythematosus (SLE). However, these small chemical drugs suffer from poor solubility, short circulating half-life and adverse side effects. One of the most effective strategies to extend the circulating time is loading drugs into nanocarriers to form nanomedicines, which is of particular interest for the treatment of cancer and viral diseases but has seldom been applied to autoimmune disorders. Herein, we successfully developed an easy but general drug delivery platform based on the new biocompatible polyhydroxyalkanoate (PHA) terpolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) (PHBVHHx). In this proof of concept study, we loaded the PHBVHHx nanocarrier with the immunosuppressant azathioprine (AZA) for SLE therapy for the first time. The AZA-PHA nanoparticles possessed ∼30% cytotoxicity and slow clearance from the kidneys. In a murine SLE model, AZA-PHA nanoparticles exhibited superior therapeutic efficacy to AZA and AZA-polylactic acid (PLA) nanoparticles without appreciable toxicity. This delivery system may provide a new and general platform for the development of nanomedicines with enhanced therapeutic efficacy and reduced side effects in SLE therapy.

Therapeutic proteins such as enzymes, hormones, and cytokines suffer from poor stability, inefficient cellular penetration, and rapid clearance from circulation. Conjugation with polymers (such as poly(ethylene glycol)) and fusion with long-acting proteins (such as albumin and Fc fragments) have been utilized to partially address the delivery issues, but these strategies require the introduction of new macromolecular substances, resulting in potential immunogenicity and toxicity. Herein, we report an easy strategy to increase the intracellular delivery efficiency and stability of proteins by combining of sortase-mediated protein cyclization and cell-penetrating peptide (CPP)-mediated intracellular delivery. We, for the first time, genetically constructed a green fluorescence protein (GFP) fused with a CPP, a transacting activator of transcription (TAT) peptide, at its C-terminus for intracellular internalization, and two sortase recognition sequences, pentaglycine and LPETG, at its N- and C-termini for cyclization. Notably, the cyclized GFP-TAT (cGFP-TAT) not only highly retained the photophysical properties of the protein but also significantly improved the in vitro stability compared with the native linear GFP (lGFP) and linear TAT peptide-fused GFP (lGFP-TAT).Moreover, cGFP-TAT showed better cellular internalization ability compared with lGFP. In C26 tumor-inoculated mice, cGFP-TAT exhibited enhanced in vivo tumor retention, with increases of 7.79- and 6.52-fold relative to lGFP and lGFP-TAT in tumor retention 3 h after intratumor administration. This proof-of-concept study has provided an easy strategy to increase the in vitro stability, intracellular delivery efficiency, and in vivo tumor retention of GFP, which would be applicable to numerous therapeutic proteins and peptides for clinical practice.

Endometrioid Endometrial Carcinoma (EEC), an estradiol-related disease, remains a serious health threat to women because of its high incidence and trend of rejuvenation. Accumulating evidence has highlighted that microRNAs (miRNAs) and messenger RNAs (mRNAs) play important roles in various biological processes involved in the pathogenesis of EEC. This study aimed to identify the potential prognostic biomarkers associated with EEC regulated by estradiol. RNA expression profiles of EEC were obtained from The Cancer Genome Atlas database (n = 408) and the original sequencing, which was performed on endometrial cancer Ishikawa cells treated with 250 nM estradiol (n = 3), 50 nM estradiol (n = 3) or control (n = 3). The TargetScan database was used to predict the target genes of prognosis-related differentially expressed miRNAs (DEMs). Subsequently, functional enrichment analysis and topological analysis were performed on the overlaps of target genes and differentially expressed mRNAs (DEGs). Kaplan-Meier analysis was used to predict prognosis-related target genes to identify prognostic biomarkers and cell population landscapes, and gene expression analysis was performed to locate prognosis-related DEGs based on single-cell transcriptomic sequencing data from the NCBI Sequence Read Archive database. Four estradiol-related DEGs were associated with prognosis, and 235 overlapping target DEGs were screened and incorporated into the functional enrichment analysis and protein-protein interaction network visualization studies. Additionally, SACS and GPR157 were identified as potential biomarkers for EEC prognosis through survival analyses. Furthermore, single-cell transcriptome data were analyzed to show changes in gene expression levels in specific cell types. This study demonstrates that miR-142-5p-SACS and miR-30a-5p-GPR157, which are regulated by estradiol, may hold promise as diagnostic and prognostic biomarkers and novel therapeutic targets for EEC.

The widespread application of protein and peptide therapeutics is hampered by their poor stability, strong immunogenicity and short half-life. However, the existing protein modification technologies require the introduction of exogenous macromolecules, resulting in inevitable immunogenicity and decreased bioactivity. Herein, we reported an easy but universal protein modification approach, self-fused concatenation (SEC), to enhance the in vitro thermal stability and in vivo tumor retention of proteins. In this proof of concept study, we successfully obtained a set of green fluorescence protein (GFP) concatemers, monomer (GFP 1), dimer (GFP 2) and trimer (GFP 3) of GFP, and systematically studied the effects of SEC on the biological activity and stability of GFP. Notably, GFP concatemers displayed remarkable improvement in in vitro bioactivity and thermal stability over the monomeric GFP. In a murine tumor model, GFP 2 and GFP 3 exhibited significantly prolonged duration, with increases of 220- and 381-fold relative to GFP 1 in tumor retention 4 h after administration. Furthermore, the biological activity, thermal stability and tumor retention can be enhanced by the concatenated number of self-fused proteins. These findings demonstrate that SEC may be a promising alternative to design advanced protein and peptide therapeutics with enhanced pharmaceutic profiles.

Background: To systematically analyze the project completion status and research characteristics in the field of Chinese Materia Medica (CMM, discipline code H32) funded by the National Natural Science Foundation of China (NSFC), thereby providing a basis for optimizing the allocation of scientific research resources and adjusting funding strategies. Methods: Based on the NSFC big database system, a retrospective analysis was conducted to descriptively analyze the research output, potential translational capacity, and research characteristics of completed projects in the CMM field funded by NSFC from 2015 to 2024. Results: From 2015 to 2024, a total of 4,574 projects in the CMM field were completed, with a compound annual growth rate (CAGR) of 3.0%. The cumulative total funding was 1.86 billion RMB, with an average of 410 KRMB per project. The average research output was 6.6 items per project, and the potential patent conversion ratio was 61%. For the first time, the 19 sub-directions within the CMM field were categorized into four echelons: double-high advantage, high-output low-conversion, low-output high-conversion, and Low-Output Low-Conversion areas. Key advantage areas included CMM pharmacodynamic substances, CMM resources, and ethnic pharmacology. Disease research primarily focused on cardiovascular and cerebrovascular diseases, tumors, metabolic diseases, and neurological disorders. Technical methodologies rapidly evolved from traditional chemical analysis to multi-omics and systems biology, while research mechanisms expanded towards multi-dimensional regulatory networks. Conclusion: NSFC funding for the CMM field has been consistently stable. The discipline demonstrates an overall trajectory characterized by deepening basic research, rapid iteration of technical methods, increasing diversification of research directions, and steadily improving translational potential. These findings provide data support and decision-making references for optimizing NSFC funding strategies in the CMM field.