PURPOSE:The optimal conversion treatment for patients with borderline resectable pancreatic cancer or locally advanced pancreatic cancer (BRPC/LAPC) remains unclear. In this study, we present the efficacy and safety results of a phase II trial evaluating tislelizumab combined with hypofractionated radiotherapy plus nab-paclitaxel/gemcitabine (THAG) in patients with BRPC/LAPC (ChiCTR2000032955, NCT05634564). PATIENTS AND METHODS:This phase II trial enrolled 56 patients with BRPC/LAPC (BRPC: 17, 30.4%; LAPC: 39, 69.6%). Participants received tislelizumab plus nab-paclitaxel/gemcitabine (AG) in 21-day cycles. Nonprogressing patients received concurrent radiotherapy during the third chemotherapy cycle. After four treatment cycles, a multidisciplinary team assessed eligibility for radical surgery. Dynamic biomolecular profiling was performed. RESULTS:Fifty-six eligible patients were enrolled. The objective response rate was 51.8% [95% confidence interval (CI), 38.0%-65.3%]. Median progression-free survival (mPFS) was 13.2 months (95% CI, 11.6-19.4 months), and median overall survival (mOS) was 21.3 months [95% CI, 18.8-not reached (NR)]. Among 30 patients who met criteria for surgical resectability, 22 patients (22/56, 39.3%) underwent radical resection, comprising nine patients with BRPC (9/17, 52.9%) and 13 patients with LAPC (13/39, 33.3%). The margin-negative resection rate reached 90.9% (95% CI, 70.8%-98.9%), and the mOS of patients who underwent surgery was 34.0 months (95% CI, 20.1-NR). Grade ≥3 adverse events (AE) occurred in 33/56 patients (58.9%). Dynamic biomarker exploration revealed that baseline IL6 level (>5 pg/mL) predicted better PFS. Moreover, circulating tumor DNA (ctDNA) status and clearance demonstrated superior survival. CONCLUSIONS:The THAG regimen as preoperative therapy showed encouraging clinical activity with a manageable safety profile. Dynamic biomarker findings reveal potential for guiding precision treatment strategies with THAG.

Calvatia lilacina (CL), a traditional Chinese medicinal fungus, has been extensively used for wound healing for centuries. However, the specific effective components of CL remain unclear. This study investigated the phytochemical composition and diabetic wound healing activities of a novel ethanol extract-derived fractions from CL (CLP2). UHPLC-TOF-MS/MS phytochemical analysis revealed 14 active components in CLP2. HPLC analysis showed that ergosterol was the most prevalent component in CLP2, with a content of 837.3 μg/mg. CLP2 significantly accelerated wound healing in db/db diabetic mice, facilitated re-epithelialization and inhibited the polarization of M1 macrophages, downregulated the transcription levels of the inflammatory cytokines, and promoted the expression of anti-inflammatory factors. In vitro tests indicated that CLP2 can promote the proliferation and migration of mouse skin fibroblasts, suppress the expression of M1 macrophage-associated pro-inflammatory cytokines, and enhance M2 polarization. Additionally, in vivo and in vitro experiments confirmed that CLP2 accelerated diabetic wound healing by suppressing M1 macrophages and promoting the polarization of macrophages towards the M2 phenotype, facilitating a faster transition from the inflammatory stage to the proliferative stage. These results highlight CLP2 as a potential therapeutic intervention for diabetic wound healing.

Hypertrophic scars (HS) represent a significant clinical challenge due to their complex pathophysiology and resistance to conventional therapies, often resulting in persistent symptoms such as itching, pain, and impaired joint mobility that compromise patients' quality of life. Current treatment modalities, including compression therapy, pharmacological agents, radiation, silicone gel, and laser therapies, have faced limitations primarily due to inadequate drug penetration into the dense fibrotic scar tissue. In this context, microneedle-mediated controlled delivery systems have emerged as a promising pharmaceutical platform to enhance localized and sustained delivery of therapeutic agents, including small-molecule drugs, biologic proteins, small interfering RNA, and living cells, directly into HS. This article critically reviews the biological and formulation-related challenges associated with transdermal delivery in scar tissue and highlights recent innovations in microneedle design, material selection, and drug-loading techniques tailored for controlled release applications. Furthermore, it discusses the integration of proteins and cell-based therapies within microneedle platforms and their potential to modulate scar remodeling and inflammation. By addressing current limitations and exploring cutting-edge technologies, this review article aims to guide the development of effective microneedle-mediated strategies for pharmaceutical intervention in hypertrophic scar management.

Triple-negative breast cancer (TNBC) is the most challenging breast cancer subtype to treat due to the absence of effective targeted therapies. In this study, we demonstrate that elevated expression of microtubule affinity-regulating kinase 2 (MARK2), but not other MARK family members (MARK1, MARK3, and MARK4), correlates with poor prognosis in TNBC patients. Silencing MARK2 impairs TNBC progression via inhibition of mutant p53 (mutp53) signaling. In contrast, silencing any of the other three MARKs either enhances or does not affect TNBC cell growth or migration and has no impact on mutp53 expression. Notably, direct knockdown of mutp53 recapitulates the effects of MARK2 ablation in TNBC cells, further supporting a functional linkage. Moreover, ectopic expression of either wild-type (WT) MARK2 or its kinase-dead (KD) mutant enhances mutp53 signaling and promotes TNBC progression; however, MARK2 overexpression does not alter wild-type p53 (wtp53) expression or cell growth in luminal breast cancer cells. Significant inverse correlations are also observed between the expression levels of MARK2, THBS1, or HBEGF (two direct target genes of mutp53) and both overall and disease-free survival in TNBC patients harboring mutTP53, whereas no such association exists between MARK2 and survival in breast cancer subtypes expressing wtTP53. MARK2 is predominantly localized in the nucleus of TNBC cells, where it interacts with and stabilizes mutp53 through its UBA and Spacer domains. Consistent with this, MARK2-ΔUBA or MARK2-ΔSpacer mutant proteins fail to bind mutp53 or sustain its signaling, thereby acting as dominant-negative inhibitors that suppress TNBC progression. Collectively, our findings indicate that suppressing MARK2 expression, rather than inhibiting its kinase activity, may represent an effective therapeutic strategy for TNBC with mutTP53.

As an important barrier tissue, the intestinal mucosa is responsible for regulating immunity, absorbing nutrients, and maintaining microbial homeostasis. Structural damage or functional impairment of the mucosa not only leads to a series of mucosa-related diseases but also may cause pathological changes in extramucosal organs. Given the central role of mucosal dysfunction in the onset and progression of various diseases, effectively regulating mucosal function and improving drug delivery efficiency are critical for improving clinical outcomes in gastroenterology. This article systematically reviews the structural characteristics and main functions of the intestinal mucosa, and analyzes the manifestations and underlying mechanisms of mucosal dysfunction. Based on this, we summarize recent advances in barrier enhancement, precise tissue and cell-level targeting, and microenvironment-responsive delivery strategies, and further elaborate on the latest progress in bio-inspired nanosystems for the treatment of gastrointestinal diseases. Finally, this article discusses the clinical trial progress, current challenges, and future development prospects of artificial intelligence optimization technology in the regulation of intestinal mucosa and drug delivery, aiming to provide theoretical basis and research references for in-depth studies on the regulation of intestinal mucosa function and the clinical application of related delivery systems.

Inflammation plays a vital role in the initiation and progression of hepatocellular carcinoma (HCC). Whether the inflammatory state of HCC peripheral liver tissue also affects tumor progression remains unclear. The medical data of HCC patients at Nanjing Drum Tower Hospital was retrospectively reviewed. Patients were divided into two groups on different inflammation grades in the peritumoral liver tissue: G 0–1 and G ≥ 2. As a preliminary exploration, RNA-seq was conducted on a limited cohort of six HCC tissues (three per group). Gene Set Enrichment Analysis (GSEA) was conducted to detect differential activation of Hallmarks pathways. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses were conducted on differentially expressed genes between these two groups to elucidate enriched pathways and biological processes. Integrated machine learning algorithms and multivariate Cox regression analysis were used for prognostic gene screening and model establishment. 163 patients were included in the G 0–1 group and 207 patients in the G ≥ 2 group. More significant liver fibrosis, as well as severe coagulation and hepatic function impairment, were related to the G ≥ 2 group. Inflammatory genes, hepatic fibrosis-related genes, and Hallmarks pathways INFLAMMATORY_RESPONSE were significantly upregulated in the G ≥ 2 group. Conversely, coagulation factors, complement molecules, fibrinolysis-related molecules, hepatic metabolism-related genes, and Hallmarks pathways COAGULATION and BILE_ACID_METABOLISM were markedly upregulated in the G 0–1 group. KEGG and GO enrichment analyses yielded results consistent with above findings. The established model based on seven inflammatory-related genes performed well in prognostic prediction and risk stratification across multiple datasets. HCC patients with G ≥ 2 had an inflammatory and fibrotic phenotype with a worse prognosis, while patients with G 0–1 retained better coagulation and liver metabolic functions.

Background & Aims Intrahepatic cholangiocarcinoma (iCCA) exhibits profound heterogeneity, complicating its clinical management. This study aims to define the molecular features underlying its histological subtypes to better understand their biology and identify potential therapeutic opportunities. Methods We performed an integrated multi-omics analysis on a cohort of 283 histologically defined iCCA samples, utilizing targeted DNA sequencing (n=207) and transcriptomic profiling (n=281). Results Cholangiolocarcinoma (64/283, 22.6%), conventional small-duct-type (SD-type, 124/283, 43.8%), and large-duct-type (LD-type, 95/283, 33.6%) iCCA exhibited distinct molecular features. Cholangiolocarcinoma was characterized by lower frequencies of KRAS and TP53 mutations, enrichment of oxidative phosphorylation-related transcriptional programs, and a relatively immune-cold microenvironment with reduced immune-related signatures and tertiary lymphoid structure (TLS)-related scores. In contrast, LD-type tumors exhibited higher KRAS and TP53 mutation rates and more aggressive features, while SD-type tumors displayed intermediate characteristics. Subtype-dependent associations were observed between metabolic pathway activity and immune features. In an independent validation cohort (n=150), cholangiolocarcinoma was associated with favorable overall survival. Conclusions This study supports cholangiolocarcinoma as a biologically distinct subtype of iCCA and highlights subtype-specific genomic, metabolic, and immune features. These findings provide a refined framework for histomolecular classification of iCCA and may inform future subtype-aware research. Impact and Implications This multi-omics study provides an integrated molecular characterization of the three major histological subtypes of intrahepatic cholangiocarcinoma (iCCA), linking histopathologic classification with distinct genomic, transcriptomic, immune, and metabolic features. Our findings support cholangiolocarcinoma (CLC) as a molecularly distinct subtype, while highlighting divergent biological programs across CLC, small-duct type, and large-duct type tumors. Rather than relying on morphology alone, this framework provides a biologically informed basis for subtype-specific investigation of iCCA. These findings establish a molecular framework for future subtype-aware translational studies in iCCA and provide a resource for generating biologically informed hypotheses regarding subtype-specific therapeutic vulnerabilities. Further validation in independent cohorts and functional studies will be required before clinical translation.

End-stage liver disease (ESLD) covers the end-stage of acute and chronic liver diseases, mainly involving decompensated cirrhosis, various types of liver failure, and advanced liver cancer. Hepatocyte transplantation has shown promise in treating ESLD, but its clinical application is hampered by the shortage of donor hepatocytes. Cell therapy, an emerging effective treatment for ESLD, faces the same limitation due to scarce hepatocyte availability. Hepatocyte-like cells (HLCs), which are terminally differentiated cells, can be induced from both stem cells and somatic cells. As HLCs exhibit the morphology and function of primary hepatocytes, they offer a promising supplementary source of hepatocytes for cell therapy. First, we provide a background for the differentiation and maturation of primary hepatocytes. Subsequently, based on the current insights into the molecular pathways that regulate hepatocyte differentiation in vivo, we describe a strategy for establishing HLC derived from either stem cells or somatic cells. The key characteristics of these HLCs are also detailed. Furthermore, HLC offers therapeutic potential for liver failure, and HLC-based liver organoids and bioartificial liver systems have demonstrated the ability to provide liver functions, offering an innovative approach to treating various types of ESLD. Despite their promise, challenges such as efficiency in differentiation and functional maturation need to be addressed to improve the clinical application of HLCs. This review discusses these advancements and outlines the therapeutic potential and current challenges of HLC therapy for ESLD.

Compound Nanxing Zhitong Plaster (CNZP) is a topical formulation widely used for rheumatoid arthritis (RA), yet its pharmacodynamic material basis is not fully understood. We established a strategy integrating chemical analysis, network pharmacology, and multi-compartment transdermal pharmacokinetics. Comprehensive chemical profiling by UPLC-Q-TOF-MS identified 100 constituents. Network pharmacology screened 15 core bioactive components. A validated UPLC-QqQ-MS/MS method was developed to simultaneously quantify eight components in the stratum corneum (SC) and viable epidermis plus dermis (VD) and four systemically absorbed components in plasma after transdermal administration, elucidating the formulation's in vivo behavior. Pharmacokinetics revealed structure-dependent transdermal patterns: Phenylpropanoids and phthalides showed rapid absorption and elimination, while organic acids, alkaloids, and flavonoids exhibited reservoir effects and slow release in the SC, contributing to a "fast-slow synergistic" mechanism. Given its high transdermal permeability, notable local and systemic exposure, and established anti-inflammatory evidence, ferulic acid is proposed as a potential quality marker for CNZP. This study provides evidence for the in vivo behavior of topical Chinese medicines and suggests methods for quality control and active substance research.

Anal fistula is one of the most common and frequently occurring diseases in the anorectal department. Calvatia lilacina spore (CLS) has been applied for wound treatment with a long history as a traditional Chinese medicine (TCM). However, the mechanism of CLS to treat postoperative wound of anal fistula remains unclear. The present study aims to investigate the efficacy and mechanism of CLS in promoting anal fistula wound healing from the perspective of regulating the interaction between macrophages and fibroblasts. Twenty patients who received anal surgery were recruited in Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine. We presented a single-cell atlas of granulation tissue, comparing samples with and without CLS treatment, utilizing single-cell RNA sequencing. The pharmacological effects and mechanism of CLS on anal fistula wound were assessed using elisa, Immunohistochemistry (IHC) staining, western blot, Immunofluorescence (IF) staining, flow cytometry assays and cell co-culture. The CLS had a uniform particle size and contained components mainly including proteins, steroids, polysaccharides and polyphenols. CLS reduced the expression levels of Tumor Necrosis Factor-alpha (TNF-α) and increased the expression levels of Vascular Endothelial Growth Factor (VEGF) and Collagen Type I Alpha 1 (COL1A1) in the granulation tissue. The single-cell sequencing revealed that the expression level of interleukin 6 (IL-6) and C-X-C Motif Chemokine Ligand 8 (CXCL-8) was increased in the IL-6+ macrophages that promoted the expression of Wiskott-Aldrich syndrome protein family member 3 (WASF3) in fibroblasts and further recruited Actin-Related Protein 2 (ACTR2), Actin-Related Protein 3 (ACTR3). Finally, CLS enhanced intercellular communication between macrophages and fibroblasts by activating the Janus Kinase 2 (JAK2)/Signal Transducer and Activator of Transcription 3 (STAT3) signaling pathway, thereby promoting mouse skin fibroblasts (MSF) migration ability. Our study objectively demonstrated the pharmacological effects of CLS in promoting the wound healing of anal fistula and investigated its mechanisms in terms of regulating the immune inflammatory process of macrophages increases signal communication with fibroblasts while promoting fibroblast transformation.

This study reports an exceptionally rare clinical encounter of synchronous multiple primary cancers involving high-grade serous ovarian carcinoma (HGSOC) and well-differentiated appendiceal mucinous adenocarcinoma in a 73-year-old woman. Presenting with abdominal distension and complex abdominopelvic masses, the patient's ascitic fluid revealed 2 morphologically distinct malignant cell populations. Following cytoreductive surgery, histopathological and immunohistochemical (IHC) analyses confirmed a dual-origin malignancy: ovarian lesions exhibited PAX8, WT1, and mutant-pattern TP53 expression, while the appendiceal lesion demonstrated strong keratin 20 positivity. Crucially, targeted gene sequencing validated these findings by identifying mutually exclusive mutational profiles-the HGSOC harbored a TP53 c.549del mutation, whereas the well-differentiated appendiceal mucinous adenocarcinoma carried KRAS p.G12D, GNAS p.R201C, and ATR p.R1951* mutations. Beyond simple coexistence, this patient had extensive cross-metastasis and tumor-to-tumor metastasis within both primary sites and metastatic deposits, likely facilitated by mucinous ascitic dissemination and lymphovascular invasion. Such diagnostic complexity necessitates a multidimensional approach integrating histomorphology, IHC, and molecular profiling to prevent misdiagnosis. Our findings emphasize that when imaging or cytology suggests multiorigin components, clinicians should pursue thorough intraoperative exploration, multisite biopsies, and prophylactic appendectomy. Ultimately, the management of such patients requires highly individualized surgical and chemotherapeutic strategies that account for the divergent biological behaviors and therapeutic sensitivities of both HGSOC and well-differentiated appendiceal mucinous adenocarcinoma to optimize oncological outcomes.

Diabetic wound healing is severely impaired by persistent inflammation and microbial dysbiosis. The study presents a combinedfunctional ingredient system centered on an engineered thymidine-auxotrophic Escherichia coli Nissle 1917 strain (ThyA-/--EcN), whose survival and activity are sustained by the thymidine naturally present in Calvatia lilacina spore extract (CLSE). CLSE was chemically characterized by UHPLC-QTOF-MS, and its thymidine content was quantified as 4.1 mg/g by HPLC. In a full-thickness excisional wound model in diabetic db/db mice, CLSE alone accelerated wound healing, enhanced re-epithelialization and collagen deposition, and restructured the wound microbiota by increasing Escherichia-Shigella while suppressing Staphylococcus (SA). Notably, the combination of CLSE and ThyA-/--EcN significantly prolonged the colonization of the engineered probiotic in vivo compared to ThyA-/--EcN alone, an effect we hypothesize to be attributable, at least in part, to the thymidine provided by CLSE. This prolonged colonization, in combination with CLSE, boosted M2 macrophage polarization, enhanced fibroblast function, and strengthened the suppression of SA, which our in vitro data suggest may involve nutrient competition. The results demonstrate that CLSE not only directly improves wound healing but also acts as a natural thymidine delivery vehicle to unlock the full potential of ThyA-/--EcN, establishing a rationale for developing a food-derived bioactive extract-assisted live biotherapeutic strategy.

Diabetic ulcers (DUs), a severe complication of diabetes, are characterized by impaired wound healing and contribute significantly to morbidity and mortality. A key pathological driver is the persistent accumulation of neutrophil extracellular traps (NETs), which extend inflammation and tissue damage; however, appropriate therapeutic strategies to resolve NETs remain underdeveloped. We engineered a self-assembled nanocomplex, O/DNase-I, through structural and functional integration of oligomerized epigallocatechin gallate (OEGCG) and deoxyribonuclease-I (DNase-I). Its functionality was systematically evaluated in vitro and in a diabetic murine wound model using molecular and histological analyses. The O/DNase-I nanocomplex simultaneously eliminates existing NETs via DNase-I-mediated DNA hydrolysis and suppresses further NET formation through OEGCG. This synergistic action robustly cleared NETs, mitigated pro-inflammatory signaling, and critically, promoted a reparative immune microenvironment by driving M2 macrophage polarization, ultimately accelerating diabetic wound closure in vivo. This study not only validates O/DNase-I as a potent therapeutic approach for diabetic wound management but also establishes a novel supramolecular strategy for targeting dysregulated inflammation, with broad potential applications in other NET-associated pathologies.

Background: The immunologically cold nature and immunosuppressive tumor microenvironment (TME) of intrahepatic cholangiocarcinoma (ICC) contribute to its poor prognosis. This study aims to identify novel biomarkers related to prognosis and TME in ICC. Methods: We first identified the high expression of m6A reader insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) in ICC through bioinformatics screening. Subsequently, a retrospective study was conducted on 224 ICC patients who had undergone radical resection. The expression levels of IGF2BP2 and programmed death ligand 1 (PD-L1) were detected in a tissue microarray (TMA) using immunohistochemistry (IHC). The co-localization of IGF2BP2, PD-L1, programmed cell death protein 1 (PD-1), and CD8+T cells was evaluated by multiple immunofluorescence techniques. Results: IHC confirmed a significant upregulation of IGF2BP2 in tumor tissues compared with normal bile duct epithelia (p < 0.05). IGF2BP2 expression was positively correlated with PD-L1 expression (TPS R = 0.215, p = 0.016; CPS R = 0.295, p = 0.008). High IGF2BP2 expression was associated with increased PD-L1/PD-1 positivity and reduced CD8+T cell infiltration. Kaplan-Meier analysis revealed significantly worse 3-year overall survival (OS: 20.56% vs. 29.91%, p = 0.0291) and recurrence-free survival (RFS: 9.72% vs. 18.56%, p = 0.0372) in the IGF2BP2-high group. Multivariate analysis identified IGF2BP2 as an independent risk factor for both OS (HR = 1.683, p = 0.044) and RFS (HR = 1.946, p = 0.042). Conclusions: IGF2BP2, as a potential biomarker and independent prognostic factor for ICC, is associated with increased PD-L1 expression.

INTRODUCTION:Peptidyl arginine deiminase 4 (PAD4) catalyzes the deamination of arginine residues to citrulline, a post-translational modification known as citrullination, which regulates protein structure, function, and localization. Despite growing evidence connects PAD4 to cancer progression, its role in intrahepatic cholangiocarcinoma (ICC) remains largely unexplored. OBJECTIVES:This study aimed to elucidate the role and underlying mechanisms of PAD4 in the regulation of intrahepatic cholangiocarcinoma proliferation, as well as to explore its clinical relevance and therapeutic potential. METHODS:Hydrodynamic tail vein injection (HTVI) and subcutaneous xenograft models were used to evaluate the role of PAD4 in vivo. In vitro functional assays were performed to assess the effects of PAD4 on ICC cell proliferation. Putative targets were identified by RNA-seq and Co-IP/MS analyses. Molecular interactions were examined using GST pull-down, surface plasmon resonance (SPR), and RNA immunoprecipitation-PCR. The clinical relevance of PAD4 was evaluated in ICC patient samples using immunohistochemical analysis. RESULTS:PAD4 was identified as a key promoter of ICC proliferation by enhancing post-transcriptional expression of the MCM complex. Mechanistically, PAD catalytic domain of PAD4 interacted with the KH1 domain of IGF2BP2 and catalyzed its citrullination at R597, which markedly increased the affinity of IGF2BP2 for m6A-modified MCM2-7 transcripts, thereby stabilizing and elevating MCM mRNAs expression. Clinically, high PAD4 expression correlated with poor prognosis in ICC patients, while PAD4 and IGF2BP2 co-expression predicted worse outcomes. In vivo, combined inhibition of PAD4 and IGF2BP2 synergistically suppressed ICC growth. CONCLUSION:Our findings identify a tumor-intrinsic PAD4-IGF2BP2 axis that drives ICC progression through citrullination-dependent stabilization of MCM transcripts. Dual targeting of PAD4 and IGF2BP2 represents a promising therapeutic strategy for ICC.

Background Neural invasion (NI) is a hallmark feature of pancreatic ductal adenocarcinoma (PDAC), detected in up to 90% of intrapancreatic specimens. However, its binary presence alone has limited prognostic value. This study aimed to refine the prognostic assessment of NI in PDAC through detailed analysis of intrapancreatic NI (IPNI) and extrapancreatic perineural invasion (EPNI). Methods Patients undergoing radical pancreatectomy for PDAC between July 2018 and December 2022 were enrolled and divided into training and internal test cohorts. IPNI was characterized using binary classification, enumeration of invaded nerve fascicles, and a novel NI severity score. EPNI was diagnosed preoperatively on CT. Cox regression analysis identified parameters associated with poor prognosis in the training cohort, which were subsequently validated in the internal test cohort. Results Among 195 patients, the overall detection rates of IPNI and EPNI were 97.4% and 42.0%, respectively. EPNI (p = 0.01), pathological N2 stage (p = 0.001), and NI severity score (p = 0.04) were independently associated with shorter recurrence-free survival (RFS). For overall survival (OS), significant independent prognostic factors included increased age (p = 0.009), EPNI (p = 0.001), pathological N2 stage (p = 0.003), poor tumor differentiation (p = 0.022), and NI severity score (p = 0.032). A NI severity score >10.5 was identified as the optimal cutoff for stratifying patients with different prognoses. Kaplan-Meier analysis demonstrated that NI severity scores >10.5 were associated with significantly poorer RFS (10 vs. 24 months, p = 0.001) and OS (19 vs. 27 months, p = 0.001). Similarly, EPNI was associated with shorter RFS (8 vs. 18.5 months, p = 0.001) and OS (17 vs. 25 months, p = 0.001). These findings were validated in the internal test cohort. Furthermore, NI severity scores >10.5 were significantly associated with early recurrence within 6 months post-pancreatectomy. Conclusions EPNI and an NI severity score exceeding 10.5 are significant independent prognostic indicators in resectable PDAC and can serve as predictors of tumor recurrence following detailed pathological assessment. Registration number No. 2023-161-01, approved by the Ethics Committee of the Drum Tower Hospital, affiliated with Nanjing University Medical School.

Upper limb spasticity is a common and disabling complication of stroke. Botulinum toxin type A (BoNT-A) is widely used for focal spasticity treatment, but naturally derived products may present limitations related to immunogenicity and manufacturing variability. Recombinant botulinum toxin type A, produced by genetic engineering without complexing proteins, may provide improved product consistency. This Ib/II study evaluated the safety, tolerability, and preliminary efficacy of recombinant botulinum toxin type A in adults with post-stroke upper limb spasticity.This multicenter, seamless Ib/II clinical study included an open-label dose-escalation Ib phase and a randomized, double-blind, placebo-controlled II phase. Adult patients with post-stroke upper limb spasticity received a single intramuscular injection of recombinant botulinum toxin type A or placebo. The primary endpoint in Phase II was the change from baseline in the Modified Ashworth Scale (MAS) score of the primary target muscle group at Week 4. Secondary endpoints included MAS and Tardieu scale changes in individual muscle groups, Disability Assessment Scale (DAS), Physician's Global Assessment (PGA), and immunogenicity.The Ib phase showed improvements in MAS, DAS, and PGA, indicating an early efficacy signal. In Phase II, recombinant botulinum toxin type A produced a significant reduction in MAS score of the primary target muscle group at Week 4 compared with placebo, with effects sustained through Week 12. At Week 4, the PGA score in the Eveotox® group showed a statistically significant improvement compared with the placebo group. While MAS and PGA scores showed significant improvement, DAS functional scores did not differ statistically from the placebo group at week 4. The treatment was generally well tolerated, and low incidence of antibodies were observed.Recombinant botulinum toxin type A was safe and effective in reducing post-stroke upper limb spasticity after a single administration. These results support further Phase III clinical evaluation.

Predicting skin penetration enhancers (PEs) from numerous terpenes is of interest for the development of the transdermal drug delivery system (TDDS) and remains a great challenge. In this study, the stratum corneum (SC) retention abilities of 3 sesquiterpenes and 3 monoterpenes from Alpiniae Officinarum Rhizoma were compared. The sesquiterpenes included caryophyllene oxide (CPO), δ-cadinene, and β-caryophyllene, while the monoterpenes contained 4-terpineol, β-pinene, and 1,8-cineole. Long-chain sesquiterpenes showed much higher SC-retention abilities than short-chain monoterpenes. The SC-retention abilities of terpenes were directly correlated with their disturbance effects on skin-barrier function (r = 0.9661, p < 0.01). Molecular dynamics (MD) simulation, molecular docking, and attenuated total reflection Fourier-transform infrared (ATR-FTIR) analysis revealed that sesquiterpenes more easily bound to SC components compared with monoterpenes. Consequently, steered molecular dynamics (SMD) simulations were performed using a novel model containing both a mixed ceramide (CER) lipid membrane and keratins. This model was used to predict the SC-retention ability of terpenes and their effects on skin-barrier disturbance. Based on these results, CPO and δ-cadinene were identified as potent PE candidates. The in vivo penetration-enhancement effects of the obtained sesquiterpenes through assessing skin permeation of FITC-Dextran-4000 (FD-4k) were comparable to azone whereas the toxicity of them was significantly lower than azone. In conclusion, we established a feasible approach to identify the potent PEs from numerous terpenes by evaluating the SC-retention ability using SMD simulation.

Doxorubicin (DOX) efficacy in breast cancer is limited by dose-dependent cardiotoxicity and the development of multidrug resistance, while celastrol (Cel) suffers from poor solubility, low bioavailability and systemic toxicity. Their combination offers synergistic potential, but effective co-delivery remains challenging. We designed a protein-based nanoplatform via ferric ion-mediated biomimetic mineralization co-assembly of DOX, Cel, and human serum albumin (HSA) for enhanced antitumor therapy. HSA-Fe(III)-Cel-DOX nanoparticles were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Drug loading and encapsulation efficiencies were determined by high-performance liquid chromatography (HPLC). In vitro drug release was evaluated with or without glutathione (GSH). Cytotoxicity against 4T1 murine breast cancer cells was assessed by CCK-8 assay. The optimized HSA-Fe(III)-Cel-DOX NPs (8 mM Fe3⁺) exhibited uniform spherical morphology with an average diameter of approximately 64.6 nm and narrow size distribution (PDI = 0.103). The nanoparticles demonstrated exceptionally high drug encapsulation efficiencies (96.7 ± 1.9

Background/Aims Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.