Background: Our genomic understanding of lymphomas, a heterogeneous group of neoplasms, has grown exponentially. The latest World Health Organization (WHO) and International Consensus classifications reflect the importance of genetic assessment in the diagnosis and prognostication of and therapeutic decision making in lymphoid neoplasms. To address this clinical need for routinely available and timely testing, we aimed to validate the Ion AmpliSeq Liverpool Lymphoid Network Panel (IALLNP; Thermo Fisher Scientific, Waltham, MA, USA). Methods: We clinically validated the IALLNP on the Ion Torrent Genexus Sequencer (Thermo Fisher Scientific). The panel detects single-nucleotide variants (SNVs) and insertions/deletions (indels) in 60 clinically relevant genes. The validation set included a commercial control and 54 DNA samples covering the spectrum of clinically aggressive and indolent lymphomas. Results: After optimizing for poor coverage regions, recurrent artifacts, and false-negative calls, the panel showed good performance in terms of depth of coverage, on-target reads, and uniformity. Its sensitivity for SNVs and indels at a lower limit of detection of 5% variant allele frequency (VAF) was 100%. Specificity in variant-negative samples was 100%, and the mean per-sample number of false-positive variants—which were easily identifiable and excluded upon interrogation of raw data—was 0.4. The panel demonstrated 92.8% reproducibility; however, all nonreproducible variants fell below the 5% VAF analytical threshold. Conclusions The IALLNP is an accurate and reproducible next-generation sequencing panel that delivers genetic results for lymphoid neoplasms in a clinically meaningful timeframe.

Inteins are unique protein insertion sequences capable of self-excision, enabling the covalent ligation of flanking extein peptides via amide bond formation. This process proceeds spontaneously without requiring external enzymes, cofactors, or chemical reagents, granting inteins exceptional biocompatibility and traceless performance in protein engineering applications. Split inteins represent a specialized and versatile subclass whose splicing domains are encoded by two separate gene fragments rather than a single continuous open reading frame. These fragments, known as the N-terminal (IntN) and C-terminal (IntC) split inteins, associate through non-covalent interactions including hydrophobic forces, hydrogen bonds, and van der Waals forces to assemble into an active three-dimensional structure, which then drives efficient extein ligation and enables protein trans-splicing. Protein trans-splicing mediated by split inteins has become a cornerstone for traceless protein ligation owing to its high specificity and irreversibility, fundamentally reshaping strategies for protein modification, assembly, and functional regulation. Compared with traditional chemical ligation methods, split intein systems require no complex chemical derivatization of peptide fragments and can operate efficiently at micromolar concentrations under physiological conditions, thus avoiding structural and functional damage caused by organic reagents. In contrast to enzymatic ligation tools such as sortase, split inteins eliminate the need for additional enzymes or cofactors, simplifying reaction systems, reducing costs, and minimizing non-specific side products. These distinctive advantages render split inteins highly promising for applications in chemical biology, synthetic biology, and biopharmaceutical development. In recent years, deepened mechanistic understanding has established structure-guided rational design as the primary approach to overcoming key limitations of split inteins, including intrinsic aggregation propensity, strict extein sequence dependence, and limited splicing efficiency. Bioinformatic tools have been used to identify aggregation-prone regions in the IntN fragment, and site-directed mutagenesis of hydrophobic residues, relocation of split sites, or removal of misfolding-prone sequences has substantially reduced in vitro aggregation and improved soluble expression and assembly activity. Rational engineering of catalytic residues and adjacent flexible loops has relaxed strict amino acid preferences at extein junctions, enhancing sequence tolerance and reducing the risk of functional impairment in target proteins. Consensus design based on multiple sequence alignments has yielded ultra-fast splicing variants such as Cfa DnaE and Cat-TerL, which exhibit significantly accelerated kinetics and improved tolerance to denaturing conditions. Meanwhile, advances in structural biology have further clarified the conformational dynamics and catalytic mechanisms of splicing, supporting the precise design of high-performance intein modules. On this basis, electrostatic interaction tuning and metagenomic screening have yielded multiple mutually orthogonal split intein pairs, enabling selective multi-fragment protein ligation and providing new routes for the efficient synthesis of large multi-domain functional proteins. With these engineered split inteins offering continuously improved performance and expanded applicability, protein trans-splicing has been widely applied in numerous cutting-edge areas of protein research and biomedicine. In gene delivery, split intein-based systems overcome the packaging limit of adeno-associated viral vectors, enabling the accurate reconstitution of large therapeutic proteins and base editors in target cells, thereby enhancing the efficacy and scope of gene therapy for genetic diseases. In internal protein sequence editing, split inteins mediate precise sequence replacement and modification in flexible regions or loops of target proteins, without the need for complex multi-step ligation and protein refolding involved in traditional protein semisynthesis. In protein-protein interaction studies, intein-mediated splicing covalently captures transient and weak intracellular complexes, enabling sensitive, high-throughput interaction detection and drug screening. In synthetic biology, conditionally controllable splicing systems support the construction of diverse intracellular and cell-surface biological logic gates for the precise regulation of cellular behavior. In mechanistic biochemical research, split inteins enable photocatalytic proximity labeling and site-specific tagging, allowing the preparation of homogeneous protein samples carrying precise post-translational modifications such as ubiquitination and polyglutamylation for chromatin interactome analysis and epigenetic studies. Moreover, covalent trapping strategies using split inteins stabilize transient enzymatic intermediates, providing unprecedented insights into molecular mechanisms such as nucleosome ubiquitination that are difficult to elucidate using conventional methods. This review systematically summarizes key technological advances in split inteins over the past decade, highlighting engineering strategies, mechanistic insights, and the development of orthogonal components. It comprehensively surveys emerging applications at the frontiers of protein research, analyzes current core challenges, and proposes future directions, particularly emphasizing artificial intelligence-driven de novo design and novel splicing pathways to break existing technical bottlenecks. By enabling traceless, efficient, and versatile protein manipulation, split inteins continue to serve as indispensable tools that drive innovation in protein engineering and fundamental life science research.

Objective To explore the efficacy of ¹⁸F-fluorodeoxyglucose positron emission tomography/computed tomography (¹⁸F-FDG PET/CT) in distinguishing the properties of abnormal lymph nodes and its role in assessing the prognosis of patients with related diseases, and to provide more precise evidence for clinical diagnosis and treatment regimen formulation. Methods The clinical data of 97 patients with malignant tumors diagnosed with abnormal lymph nodes at Wuxi People’s Hospital Affiliated to Nanjing Medical University between April 2024 and June 2025 were retrospectively analyzed. Patients were divided into a malignant lymph node group (n=47) and a benign lymph node group (n=50) based on pathological results. PET/CT metabolic parameters [maximum standardized uptake value (SUV_max), mean standardized uptake value (SUV_mean), metabolic tumor volume (MTV), and total lesion glycolysis (TLG)] were compared between the two groups of patients. Receiver operating characteristic curves were used to evaluate the diagnostic value of ¹⁸F-FDG PET/CT parameters for abnormal lymph nodes. Progression-free survival was compared among patients in the malignant lymph node group stratified by different levels of SUV_max, SUV_mean, MTV, and TLG. Results SUV_max, SUV_mean, MTV, and TLG were significantly higher in the malignant lymph node group than in the benign lymph node group (P < 0.05). The areas under the receiver operating characteristic curves for predicting malignant lymph nodes with SUV_max, SUV_mean, MTV, and TLG were 0.761, 0.855, 0.860, and 0.792, with sensitivities of 74.47%, 82.98%, 74.47%, and 70.21%, and specificities of 76.00%, 76.00%, 90.00%, and 82.00%, respectively (P < 0.05). Comparison of progression-free survival in patients with malignant lymph nodes with different ¹⁸F-FDG PET/CT parameter levels revealed that patients with lower levels of SUV_max, SUV_mean, MTV, and TLG had significantly longer progression-free survival than those with higher levels (Log-rank χ²=4.297, P=0.038; Log-rank χ²=6.569, P=0.010; Log-rank χ²=5.970, P=0.015; and Log-rank χ²=7.422, P=0.006, respectively). Conclusion ¹⁸F-FDG PET/CT metabolic parameters (SUV_max, SUV_mean, MTV, and TLG) demonstrate high efficacy in the differential diagnosis of benign and malignant abnormal lymph nodes and prognostic evaluation, providing significant guidance for clinical treatment decisions.

Objective To explore the efficacy of ¹⁸F-fluorodeoxyglucose positron emission tomography/computed tomography (¹⁸F-FDG PET/CT) in distinguishing the properties of abnormal lymph nodes and its role in assessing the prognosis of patients with related diseases, and to provide more precise evidence for clinical diagnosis and treatment regimen formulation. Methods The clinical data of 97 patients with malignant tumors diagnosed with abnormal lymph nodes at Wuxi People’s Hospital Affiliated to Nanjing Medical University between April 2024 and June 2025 were retrospectively analyzed. Patients were divided into a malignant lymph node group (n=47) and a benign lymph node group (n=50) based on pathological results. PET/CT metabolic parameters [maximum standardized uptake value (SUV_max), mean standardized uptake value (SUV_mean), metabolic tumor volume (MTV), and total lesion glycolysis (TLG)] were compared between the two groups of patients. Receiver operating characteristic curves were used to evaluate the diagnostic value of ¹⁸F-FDG PET/CT parameters for abnormal lymph nodes. Progression-free survival was compared among patients in the malignant lymph node group stratified by different levels of SUV_max, SUV_mean, MTV, and TLG. Results SUV_max, SUV_mean, MTV, and TLG were significantly higher in the malignant lymph node group than in the benign lymph node group (P < 0.05). The areas under the receiver operating characteristic curves for predicting malignant lymph nodes with SUV_max, SUV_mean, MTV, and TLG were 0.761, 0.855, 0.860, and 0.792, with sensitivities of 74.47%, 82.98%, 74.47%, and 70.21%, and specificities of 76.00%, 76.00%, 90.00%, and 82.00%, respectively (P < 0.05). Comparison of progression-free survival in patients with malignant lymph nodes with different ¹⁸F-FDG PET/CT parameter levels revealed that patients with lower levels of SUV_max, SUV_mean, MTV, and TLG had significantly longer progression-free survival than those with higher levels (Log-rank χ²=4.297, P=0.038; Log-rank χ²=6.569, P=0.010; Log-rank χ²=5.970, P=0.015; and Log-rank χ²=7.422, P=0.006, respectively). Conclusion ¹⁸F-FDG PET/CT metabolic parameters (SUV_max, SUV_mean, MTV, and TLG) demonstrate high efficacy in the differential diagnosis of benign and malignant abnormal lymph nodes and prognostic evaluation, providing significant guidance for clinical treatment decisions.

Objective To explore the efficacy of ¹⁸F-fluorodeoxyglucose positron emission tomography/computed tomography (¹⁸F-FDG PET/CT) in distinguishing the properties of abnormal lymph nodes and its role in assessing the prognosis of patients with related diseases, and to provide more precise evidence for clinical diagnosis and treatment regimen formulation. Methods The clinical data of 97 patients with malignant tumors diagnosed with abnormal lymph nodes at Wuxi People’s Hospital Affiliated to Nanjing Medical University between April 2024 and June 2025 were retrospectively analyzed. Patients were divided into a malignant lymph node group (n=47) and a benign lymph node group (n=50) based on pathological results. PET/CT metabolic parameters [maximum standardized uptake value (SUV_max), mean standardized uptake value (SUV_mean), metabolic tumor volume (MTV), and total lesion glycolysis (TLG)] were compared between the two groups of patients. Receiver operating characteristic curves were used to evaluate the diagnostic value of ¹⁸F-FDG PET/CT parameters for abnormal lymph nodes. Progression-free survival was compared among patients in the malignant lymph node group stratified by different levels of SUV_max, SUV_mean, MTV, and TLG. Results SUV_max, SUV_mean, MTV, and TLG were significantly higher in the malignant lymph node group than in the benign lymph node group (P < 0.05). The areas under the receiver operating characteristic curves for predicting malignant lymph nodes with SUV_max, SUV_mean, MTV, and TLG were 0.761, 0.855, 0.860, and 0.792, with sensitivities of 74.47%, 82.98%, 74.47%, and 70.21%, and specificities of 76.00%, 76.00%, 90.00%, and 82.00%, respectively (P < 0.05). Comparison of progression-free survival in patients with malignant lymph nodes with different ¹⁸F-FDG PET/CT parameter levels revealed that patients with lower levels of SUV_max, SUV_mean, MTV, and TLG had significantly longer progression-free survival than those with higher levels (Log-rank χ²=4.297, P=0.038; Log-rank χ²=6.569, P=0.010; Log-rank χ²=5.970, P=0.015; and Log-rank χ²=7.422, P=0.006, respectively). Conclusion ¹⁸F-FDG PET/CT metabolic parameters (SUV_max, SUV_mean, MTV, and TLG) demonstrate high efficacy in the differential diagnosis of benign and malignant abnormal lymph nodes and prognostic evaluation, providing significant guidance for clinical treatment decisions.

Background Intrauterine growth restriction (IUGR) is closely associated with placental dysfunction. However, the effects of preconception co-exposure to β-cypermethrin (β-CYP) and emamectin benzoate (EMB) on fetal growth and placental function remain unclear. Objective To investigate the effects of prenatal exposure to β-CYP and EMB on embryonic development in female rats and to elucidate the mechanisms by which these substances cause IUGR from the perspectives of placental structure, progesterone synthesis, and mitochondrial autophagy. Methods Forty-eight specific pathogen-free (SPF) adult female Sprague-Dawley (SD) rats were randomly assigned to a control group, a β-CYP group, an EMB group, and a co-exposure group (n=12 per group). A whole-body dynamic inhalation exposure system was used to simulate greenhouse exposure. Rats in the control group were exposed to clean air, whereas those in the exposure groups were exposed to β-CYP (55 mg·m⁻³), EMB (7.8 mg·m⁻³), or their combination for 2 h per day for 90 consecutive days. After exposure, ovarian and serum estradiol (E₂) levels were determined in six rats from each group, while the remaining six rats were mated. Of the mated rats, three per group were sacrificed on gestational day 18, and the other remaining three were allowed to deliver naturally. Pregnancy outcomes and fetal development were evaluated. Placental morphology, progesterone (Prog) levels, progesterone synthesis-related factors, oxidative stress, mitochondrial function, and mitophagy-related indicators were subsequently assessed. Results Compared with the control group, fetal body weight on gestational day 18 and neonatal body weight were significantly decreased, whereas the incidence of IUGR was significantly increased in the β-CYP, EMB, and co-exposure groups (P<0.05). The EMB and co-exposure groups exhibited an increased proportion of the placental junctional zone thickness to total placental thickness and a reduced proportion of the labyrinth zone (P<0.05). Ovarian E₂ and placental Prog levels were decreased in all three exposure groups. In addition, the placental expression of cytochrome P450 family 11 subfamily A member1(CYP11A1)、3β-hydroxysteroid dehydrogenase type 1(HSD3B1)、progesterone receptor(PgR) was downregulated, catalase (CAT) activity and adenosine triphosphate (ATP) content were reduced, and lactate dehydrogenase (LDH) activity was increased in all exposure groups (P < 0.05). Furthermore, the expression of translocase of outer mitochondrial membrane 2 was decreased, whereas the expression levels of microtubule-associated protein 1 light chain 3 (LC3), PTEN-induced kinase 1 (PINK1), parkin RBR E3 ubiquitin-protein ligase (Parkin), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) were increased in all exposure groups (P<0.05). Conclusion Prenatal exposure to β-CYP and EMB can cause structural damage to the rat placenta, affect the expression of markers related to progesterone synthesis, and is accompanied by oxidative stress, mitochondrial dysfunction, and alterations in PINK1/Parkin-mediated mitochondrial autophagy, ultimately affecting the intrauterine growth and development of the fetus.

Objective::There is limited evidence regarding the choice of P2Y 12 receptor inhibitors as a component of dual antiplatelet therapy in patients with left main (LM) disease undergoing percutaneous coronary intervention (PCI). This study aimed to evaluate long-term clinical outcomes of ticagrelor- vs. clopidogrel-based dual antiplatelet therapy strategy in acute coronary syndrome (ACS) patients undergoing LM PCI. Methods::This is a post-hoc analysis from a prospective, single-center, real-world PCI registry. A total of 1,163 patients discharged post-ACS who underwent LM PCI and received ticagrelor or clopidogrel between March 2016 and March 2019 were included in the study. The primary endpoint was ischemic events at 12 months, including cardiac death, myocardial infarction, or stroke. Secondary outcomes included all-cause death and Bleeding Academic Research Consortium types 2, 3, and 5, and types 3 and 5 bleeding. Propensity score matching was used to adjust for bias due to confounders between the 2 groups.Results::The ticagrelor and clopidogrel groups comprised 529 (45.49%) and 634 (54.51%) patients, respectively. During the follow-up period, the rate of ischemic events was significantly lower with ticagrelor than with clopidogrel before (1.32% (7/529) vs. 3.63% (23/634), P = 0.013,6) and after propensity score matching (1.41% (6/425) vs. 4.00% (17/425), P = 0.020,1). The rates of all-cause death, Bleeding Academic Research Consortium-defined type 2, 3, and 5 bleeding, and type 3 and 5 bleeding were similar between the ticagrelor group and clopidogrel group before or after propensity score matching adjustment (all P > 0.05). Conclusion::Among patients with ACS undergoing LM PCI, ticagrelor use was associated with ischemic events benefit without excessive risk of bleeding at 12 months compared with clopidogrel.

OBJECTIVE: Peritoneal fibrosis (PF) is an adverse event that occurs during long-term peritoneal dialysis, significantly impairing treatment efficiency and adversely affecting patient outcomes. Astragaloside IV (AS-IV), a principal active component derived from Astragalus membranaceus (Fisch.) Bunge, has exhibited anti-inflammatory and antifibrotic effects in various settings. This study aims to investigate the potential therapeutic efficacy and mechanism of AS-IV in the treatment of PF. METHODS: The PF mouse model was established by intraperitoneal injection of 4.25% peritoneal dialysis fluid (100 mL/kg). The epithelial-mesenchymal transition (EMT) of HMrSV5 cells was induced by the addition of 10 ng/mL transforming growth factor β (TGF-β). The differentially expressed genes in HMrSV5 cells treated with AS-IV were screened using transcriptome sequencing analysis. The potential targets of AS-IV were screened using network pharmacology and analyzed using molecular docking and molecular dynamics simulations. RESULTS: Administration of AS-IV at doses of 20, 40, or 80 mg/kg effectively mitigated the increase in peritoneal thickness and the development of fibrosis in mice with PF. The expression of the fibrosis marker α-smooth muscle actin in the peritoneum was significantly decreased in AS-IV-treated mice. The treatment of AS-IV (10, 20, and 40 μmol/L) significantly delayed the EMT of HMrSV5 cells induced by TGF-β, as demonstrated by the decreased number of 5-ethynyl-2'-deoxyuridine-positive cells, reduced migrated area, and decreased expression of fibrosis markers. A total of 460 differentially expressed genes were detected in AS-IV-treated HMrSV5 cells through transcriptome sequencing, with notable enrichment in the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)-AKT serine/threonine kinase 1 (AKT) signaling pathway. The reduced levels of phosphorylated PI3K (p-PI3K) and p-AKT were detected in HMrSV5 cells with AS-IV treatment. Epidermal growth factor receptor (EGFR) was predicted as a direct target of AS-IV, exhibiting strong hydrogen bond interactions. The activation of the PI3K-AKT pathway by the compound 740Y-P, and the activation of the EGFR pathway by NSC 228155 each partially counteracted the inhibitory effect of AS-IV on the EMT of HMrSV5 cells. CONCLUSION AS-IV delayed the EMT process in peritoneal mesothelial cells and slowed the progression of PF, potentially serving as a therapeutic agent for the early prevention and treatment of PF. Please cite this article as: Huang Y, Chu CL, Qiu WH, Chen JY, Cao LX, Ji SY, Zhu B, Wang GK, Shen QQ. Astragaloside IV delayed the epithelial-mesenchymal transition in peritoneal fibrosis by inhibiting the activation of EGFR and PI3K-AKT pathways. J Integr Med. 2025; 23(6):694-705.
Epithelial-Mesenchymal Transition/drug effects* ; Animals ; Saponins/pharmacology* ; Triterpenes/pharmacology* ; Mice ; Peritoneal Fibrosis/pathology* ; Proto-Oncogene Proteins c-akt/metabolism* ; ErbB Receptors/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Signal Transduction/drug effects* ; Male ; Humans ; Molecular Docking Simulation ; Cell Line ; Mice, Inbred C57BL

Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

In this study, araucarene diterpenes, characterized by a pimarene skeleton with a variably oxidized side chain at C-13, were investigated. A total of 16 araucarene diterpenoids and their derivatives were isolated from the woods of Agathis dammara, including 11 previously unreported compounds: dammaradione (1), dammarones D-G (2, 5, 14, 15), dammaric acids B-F (8-12), and dammarol (16). The structures of these new compounds were elucidated using high-resolution electrospray ionization mass spectroscopy (HR-ESI-MS) and one-dimensional/two-dimensional (1D/2D) nuclear magnetic resonance (NMR), while their absolute configurations were determined through the electronic circular dichroism (ECD) exciton chirality method and Snatzke's method. The hypoglycemic activity of all isolated compounds was evaluated using a transgenic zebrafish model, and a structure-activity relationship (SAR) analysis was conducted. Araucarone (3) and dammaric acid C (9), serving as representative compounds, demonstrated significant hypoglycemic effects on zebrafish. The primary mechanism involves the promotion of pancreatic β cell regeneration and glucose uptake. Specifically, these compounds enhance the differentiation of pancreatic endocrine precursor cells (PEP cells) into β cells in zebrafish.
Zebrafish ; Animals ; Diterpenes/isolation & purification* ; Insulin-Secreting Cells/cytology* ; Glucose/metabolism* ; Hypoglycemic Agents/isolation & purification* ; Molecular Structure ; Structure-Activity Relationship ; Plant Extracts/pharmacology* ; Regeneration/drug effects*