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.

This study aims to explore the mechanism through which Yishen Jiangtang Decoction(YSJTD) regulates endoplasmic reticulum stress(ERS)-mediated NOD-like receptor thermal protein domain associated protein 3(NLRP3) inflammasome to improve diabetic nephropathy(DN) in db/db mice. Thirty db/db mice were randomly divided into the model group, YSJTD group, ERS inhibitor 4-phenylbutyric acid(4-PBA) group, with 10 mice in each group. Additionally, 10 db/m mice were selected as the control group. The YSJTD group was orally administered YSJTD at a dose of 0.01 mL·g~(-1), the 4-PBA group was orally administered 4-PBA at a dose of 0.5 mg·g~(-1), and the control and model groups were given an equal volume of carboxylmethyl cellulose sodium. The treatments were administered once daily for 8 weeks. Food intake, water consumption, and body weight were recorded every 2 weeks. After the intervention, fasting blood glucose(FBG), glycosylated hemoglobin(HbA1c), urine microalbumin(U-mALB), 24-hour urine volume, serum creatinine(Scr), and blood urea nitrogen(BUN) were measured. Inflammatory markers interleukin-1β(IL-1β) and interleukin-18(IL-18) were detected using the enzyme-linked immunosorbent assay(ELISA). Renal pathology was assessed through hematoxylin-eosin(HE), periodic acid-Schiff(PAS), and Masson staining, and transmission electron microscopy(TEM). Western blot was used to detect the expression levels of glucose-regulated protein 78(GRP78), C/EBP homologous protein(CHOP), NLRP3, apoptosis-associated speck-like protein containing CARD(ASC), cysteinyl aspartate-specific proteinase(caspase-1), and gasdermin D(GSDMD) in kidney tissues. The results showed that compared to the control group, the model group exhibited poor general condition, increased weight and food and water intake, and significantly higher levels of FBG, HbA1c, U-mALB, kidney index, 24-hour urine volume, IL-1β, and IL-18. Compared to the model group, the YSJTD and 4-PBA groups showed improved general condition, increased body weight, decreased food intake, and lower levels of FBG, U-mALB, kidney index, 24-hour urine volume, and IL-1β. Specifically, the YSJTD group showed a significant reduction in IL-18 levels compared to the model group, while the 4-PBA group exhibited decreased water intake and HbA1c levels compared to the model group. Although there was a decreasing trend in water intake and HbA1c in the YSJTD group, the differences were not statistically significant. No significant differences were observed in BUN, Scr, and kidney weight among the groups. Renal pathology revealed that the model group exhibited more severe renal damage compared to the control group. Kidney sections from the model group showed diffuse mesangial proliferation in the glomeruli, tubular edema, tubular dilation, significant inflammatory cell infiltration in the interstitium, and increased glycogen staining and blue collagen deposition in the basement membrane. In contrast, the YSJTD and 4-PBA groups showed varying degrees of improvement in renal damage, glycogen staining, and collagen deposition, with the YSJTD group showing more significant improvements. TEM analysis indicated that the model group had extensive cytoplasmic edema, homogeneous thickening of the basement membrane, fewer foot processes, and widening of fused foot processes. In the YSJTD and 4-PBA groups, cytoplasmic swelling of renal tissues was reduced, the basement membrane remained intact and uniform, and foot process fusion improved.Western blot results indicated that compared to the control group, the model group showed upregulation of GRP78, CHOP, GSDMD, NLRP3, ASC, and caspase-1 expression. In contrast, both the YSJTD and 4-PBA groups showed downregulation of these markers compared to the model group. These findings suggest that YSJTD exerts a protective effect against DN by alleviating NLRP3 inflammasome activation through the inhibition of ERS, thereby improving the inflammatory response in db/db DN mice.
Animals ; Endoplasmic Reticulum Stress/drug effects* ; Diabetic Nephropathies/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Inflammasomes/drug effects* ; Male ; Kidney/pathology* ; Endoplasmic Reticulum Chaperone BiP ; Humans ; Interleukin-18/genetics* ; Mice, Inbred C57BL

This consensus will introduce the characteristics of fillers used in the surgical cavities of domestic nasal surgery patients based on relevant literature and expert opinions. It will also provide recommendations for the selection of cavity fillers for different nasal diseases, with chronic sinusitis as a representative example.
Humans ; Nasal Cavity/surgery* ; Nasal Surgical Procedures ; China ; Consensus ; Sinusitis/surgery* ; Dermal Fillers

Targeting T-cell is a strategy to control allogeneic response disorders, such as acute graft-versus-host disease (GVHD) which is an important cause of therapy-failure after allogeneic hematopoietic cell transplants. Free fatty acid receptor-4 (FFAR4) is a regulator of obesity but its role in T-cell and allogeneic reactions is unknown. Here, we found knockout of Ffar4 in donor T-cells in a mouse allograft model increased acute GVHD whereas the natural FFAR4 ligands and the synthetic FFAR4 agonists decreased it. FFAR4 agonist-mediated anti-acute GVHD effects depended on FFAR4-expression in donor T-cells. The FFAR4 agonist CpdA suppressed donor T-cell-mediated alloreaction by activating an aryl hydrocarbon receptor (AhR) pathway. CpdA recruited β-Arrestin2 to FFAR4 which facilitated nuclear translocation of AhR and upregulation of IL-22. The CpdA-mediated anti-acute GVHD effect was absent in mice receiving Ahr-knockout or Il22-knockout T-cells. Recipient-expressing Ffar4 was also important for the anti-acute GVHD effect of CpdA which inhibited activation of antigen presenting cells. Importantly, CpdA decreased acute GVHD in obese mice, an effect also depended on Ffar4-expression in donor T-cells and recipients. Our study shows the immunoregulatory effect of FFAR4 in T-cell, and targeting FFAR4 might be a relative option for controlling allogeneic reactions in obese patients.

Tailored lipid nanoparticles (LNPs)-mediated small interfering RNA (siRNA) nanomedicines show promise in treating liver disease, such as acute liver injury (ALI) and non-alcoholic steatohepatitis (NASH). However, constructing LNPs that address biosafety concerns, ensure efficient delivery, and target specific hepatic subcellular fractions has been challenging. To evade above obstacles, we develop three novel self-degradable "gemini-like" ionizable lipids (SS-MA, SS-DC, SS-MH) by incorporating disulfide bonds and modifying the length of ester bond and tertiary amino head. Our findings reveal that the disulfide-bond-bridged LNPs exhibit reduction-responsive drug release, improving both biosafety and siRNA delivery efficiency. Furthermore, the distance of ester bond and tertiary amino head significantly influences the LNPs' pK _a, thereby affecting endosomal escape, hemolytic efficiency, absorption capacity of ApoE, uptake efficiency of hepatocytes and liver accumulation. We also develop the modified-mannose LNPs (M-LNP) to target liver macrophages specifically. The optimized M-MH_LNP@TNFα exhibits potential in preventing ALI by decreasing tumor necrosis factor α (TNFα) levels in the macrophages, while MH_LNP@DGAT2 could treat NASH by selectively degrading diacylglycerol O-acyltransferase 2 (DGAT2) in the hepatocytes. Our findings provide new insights into developing novel highly effective and low-toxic "gemini-like" ionizable lipids for constructing LNPs, potentially achieving more effective treatment for hepatic diseases.

[Objective]To explore the alleviating effect of Yunpi Yishen Tongdu Decoction on spinal lesion and immune inflammation of ankylosing spondylitis(AS)model mice,and to provide a scientific basis for its clinical application.[Methods]A total of 42 female BALB/c mice were randomly divided into blank group,model group,western medicine group,low,medium and high dose Yunpi Yishen Tongdu Decoction groups.Except for blank group,the remaining 5 groups were injected with proteoglycans in the abdominal cavity to construct AS model.After modeling,the mice in blank group and model group were given 0.9％sodium chloride solution,the mice in western medicine group were given 3.57 mg·mL-1 celecoxib solution,and the mice in the low,medium,and high dose Yunpi Yishen Tongdu Decoction were given 0.9,1.8 and 3.6 mg·mL-1 of Yunpi Yishen Tongdu Decoction for 4 weeks,respectively.After gavage,the behavioral changes,serum inflammation cytokines,spinal lesions,and expression of receptor activator of nuclear factor-κB ligand(RANKL),receptor activator of nuclear factor-κB(RANK),and osteoclastogenesis inhibitory factor(OPG)in the spine were measured.[Results]Compared with blank group,model group had significantly increased total time in tail immobility,serum interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)level,spinal pathological score,RANKL and RANK expression(P＜0.01,P＜0.05),while the vertebral bone mineral density and OPG expression of the spine were significantly decreased(P＜0.01).After treatment with the Yunpi Yishen Tongdu Decoction,the latency time in tail immobility in AS mice was significantly reduced,along with decreased serum levels of IL-6 and TNF-α and a notable reduction in spinal pathological scores(P＜0.01,P＜0.05).Additionally,the treatment downregulated RANKL and RANK expression,upregulated OPG expression(P＜0.01,P＜0.05).[Conclusion]Yunpi Yishen Tongdu Decoction alleviates the bone destruction of AS by affecting the RANKL/RANK/OPG pathway.