ObjectivePancreatic ductal adenocarcinoma (PDAC) exhibits a limited response to current treatments due to its dense fibrotic stroma and highly immunosuppressive tumor microenvironment. In recent years, advancements in cellular immunotherapy, particularly chimeric antigen receptor macrophage (CAR-M) therapy, have offered new hope for pancreatic cancer treatment. Although CAR-M therapy demonstrates dual potential in directly killing tumor cells and remodeling the immune microenvironment, it still faces challenges such as complex in vitro preparation processes and low in vivo targeting and delivery efficiency. Therefore, developing strategies for efficient and targeted in vivo delivery of CAR genes has become crucial for overcoming current therapeutic limitations. This study aims to develop an orally administrable nano-gene delivery system for the targeted delivery of CAR genes to pancreatic tumor sites. MethodsCore nano-gene particles (PNP/pCAR) were constructed by loading plasmid DNA encoding CAR (pCAR) with cationic polypeptides (PNP). Subsequently, PNP/pCAR was surface-modified with β-glucan to prepare the targeted nanoparticles (βGlus-PNP/pCAR). The loading efficiency of PNP for pCAR was quantitatively assessed by gel retardation assay. The particle size, Zeta potential, morphology, and storage stability of PNP/pCAR were characterized using a Malvern particle size analyzer and transmission electron microscopy. At the cellular level, RAW 264.7 macrophages were selected. The cytotoxicity of PNP/pCAR was evaluated using the CCK-8 assay. The cellular uptake efficiency and lysosomal escape ability of the nanoparticles were assessed via flow cytometry and confocal microscopy. Transfection efficiency was quantitatively evaluated by detecting the expression of the reporter gene GFP using flow cytometry. At the in vivo level, an orthotopic pancreatic cancer mouse model was established. Cy7-labeled βGlus-PNP/pCAR nanoparticles were administered orally, and the fluorescence distribution in mice was dynamically monitored at 1, 2, 4, 8, and 16 h post-administration using a small animal in vivo imaging system. Forty-eight hours after oral gavage, the mice were euthanized, and pancreatic tumor tissues were collected for further analysis of intratumoral fluorescence signals using the imaging system. Additionally, βGlus-PNP/pCAR-GFP nanoparticles loaded with the reporter gene (GFP) were administered orally. Forty-eight hours post-administration, pancreatic tumor tissues were harvested to prepare frozen sections, and GFP expression was observed and analyzed under a fluorescence microscope. ResultsThe PNP carrier exhibited a high loading capacity for pCAR. The successfully prepared PNP/pCAR nanoparticles were regular spheres with a hydrodynamic diameter of approximately (120±10) nm and a Zeta potential of about +(6±1) mV. They maintained good structural stability after incubation in PBS buffer for 7 d. Cell experiments demonstrated that PNP/pCAR exhibited no significant cytotoxicity in RAW 264.7 cells while being efficiently internalized and effectively escaping lysosomal degradation. The transfection positive rate of PNP/pCAR-GFP in RAW 264.7 cells reached (25±3)%, surpassing that of Lipofectamine 2000-loaded pCAR-GFP (Lipo/pCAR-GFP), which was (20±1)%.In vivo experiments revealed that, compared to unmodified PNP/pCAR, βGlus-PNP/pCAR exhibited strongerin situ pancreatic tumor targeting ability after oral administration. Furthermore, oral administration of βGlus-PNP/pCAR-GFP resulted in significant GFP protein expression detectable within pancreatic tumor tissues. ConclusionThis study successfully constructed and validated an orally administrable, pancreatic cancer-targeting polypeptide-based nano-gene delivery system. It provides an important technological foundation in delivery systems and experimental basis for the subsequent development of in situ CAR-M-based therapeutic strategies for pancreatic cancer.

ObjectivePancreatic ductal adenocarcinoma (PDAC) exhibits a limited response to current treatments due to its dense fibrotic stroma and highly immunosuppressive tumor microenvironment. In recent years, advancements in cellular immunotherapy, particularly chimeric antigen receptor macrophage (CAR-M) therapy, have offered new hope for pancreatic cancer treatment. Although CAR-M therapy demonstrates dual potential in directly killing tumor cells and remodeling the immune microenvironment, it still faces challenges such as complex in vitro preparation processes and low in vivo targeting and delivery efficiency. Therefore, developing strategies for efficient and targeted in vivo delivery of CAR genes has become crucial for overcoming current therapeutic limitations. This study aims to develop an orally administrable nano-gene delivery system for the targeted delivery of CAR genes to pancreatic tumor sites. MethodsCore nano-gene particles (PNP/pCAR) were constructed by loading plasmid DNA encoding CAR (pCAR) with cationic polypeptides (PNP). Subsequently, PNP/pCAR was surface-modified with β-glucan to prepare the targeted nanoparticles (βGlus-PNP/pCAR). The loading efficiency of PNP for pCAR was quantitatively assessed by gel retardation assay. The particle size, Zeta potential, morphology, and storage stability of PNP/pCAR were characterized using a Malvern particle size analyzer and transmission electron microscopy. At the cellular level, RAW 264.7 macrophages were selected. The cytotoxicity of PNP/pCAR was evaluated using the CCK-8 assay. The cellular uptake efficiency and lysosomal escape ability of the nanoparticles were assessed via flow cytometry and confocal microscopy. Transfection efficiency was quantitatively evaluated by detecting the expression of the reporter gene GFP using flow cytometry. At the in vivo level, an orthotopic pancreatic cancer mouse model was established. Cy7-labeled βGlus-PNP/pCAR nanoparticles were administered orally, and the fluorescence distribution in mice was dynamically monitored at 1, 2, 4, 8, and 16 h post-administration using a small animal in vivo imaging system. Forty-eight hours after oral gavage, the mice were euthanized, and pancreatic tumor tissues were collected for further analysis of intratumoral fluorescence signals using the imaging system. Additionally, βGlus-PNP/pCAR-GFP nanoparticles loaded with the reporter gene (GFP) were administered orally. Forty-eight hours post-administration, pancreatic tumor tissues were harvested to prepare frozen sections, and GFP expression was observed and analyzed under a fluorescence microscope. ResultsThe PNP carrier exhibited a high loading capacity for pCAR. The successfully prepared PNP/pCAR nanoparticles were regular spheres with a hydrodynamic diameter of approximately (120±10) nm and a Zeta potential of about +(6±1) mV. They maintained good structural stability after incubation in PBS buffer for 7 d. Cell experiments demonstrated that PNP/pCAR exhibited no significant cytotoxicity in RAW 264.7 cells while being efficiently internalized and effectively escaping lysosomal degradation. The transfection positive rate of PNP/pCAR-GFP in RAW 264.7 cells reached (25±3)%, surpassing that of Lipofectamine 2000-loaded pCAR-GFP (Lipo/pCAR-GFP), which was (20±1)%.In vivo experiments revealed that, compared to unmodified PNP/pCAR, βGlus-PNP/pCAR exhibited strongerin situ pancreatic tumor targeting ability after oral administration. Furthermore, oral administration of βGlus-PNP/pCAR-GFP resulted in significant GFP protein expression detectable within pancreatic tumor tissues. ConclusionThis study successfully constructed and validated an orally administrable, pancreatic cancer-targeting polypeptide-based nano-gene delivery system. It provides an important technological foundation in delivery systems and experimental basis for the subsequent development of in situ CAR-M-based therapeutic strategies for pancreatic cancer.

BACKGROUND:Gait movements are one of the important characteristics exhibited by athletes during exercise,reflecting their physical condition and athletic ability.In a state of fatigue,athletes may exhibit abnormal gait movements,such as reduced stride and body shaking,which can cause harm to their bodies.OBJECTIVE:To promote technological progress in the field of sports science by applying advanced algorithms and data analysis techniques to the training practice of athletes,so as to further improve the recognition accuracy of gait movements under sports fatigue.METHODS:A gait recognition method for athletes in fatigue state was based on fish swarm algorithm.By utilizing the normalized autocorrelation function and the principle of motion energy distribution,a single cycle gait energy map of athletes was obtained.Singular value decomposition was used to transform the image to highlight visual differences,generating a gait energy map of athletes.A convolutional neural network was used to construct a gait action recognition model,and the parameters of the model were solved using the fish swarm algorithm to improve the accuracy and efficiency of fatigue gait action recognition.RESULTS AND CONCLUSION:(1)The fish swarm algorithm had a small loss value in gait action recognition,and could accurately and quickly identify the gait actions of athletes,and dynamically monitor their physical fatigue.(2)The research on fatigue gait recognition of athletes based on fish swarm algorithm could effectively identify the gait movements of athletes in fatigue state and achieve accurate capture of subtle gait changes.(3)The system stability of this method is good,which can reduce the volatility of experimental test results and improve recognition efficiency,can more effectively manage sports fatigue and prevent sports injuries.In addition,when the gait characteristics of normal people change significantly,the system can give an early warning,indicating that the individual may be in a state of fatigue and need to rest or adjust the intensity of activity.

OBJECTIVE: Ulcerative colitis is closely associated with intestinal stem cell (ISC) loss and impaired intestinal mucus barrier. Sinisan (SNS), a compound Chinese herbal medicine, has a long history in the treatment of intestinal dysfunction, yet whether SNS can relieve acute experimental colitis by modulating ISC proliferation and secretory cell differentiation has not been studied. Our study tested the effect of SNS against acute colitis and focused on the mechanisms involving intestinal barrier recovery. METHODS: Network pharmacology analysis and blood entry component analysis of SNS were used to explore the underlying mechanism by which SNS affects the acute dextran sulfate sodium (DSS)-induced murine colitis model. RNA-sequencing was used to demonstrate the mechanism. Further, reverse transcription-quantitative polymerase chain reaction, immunofluorescence staining, and alcian blue and periodic acid-Schiff staining were performed in vivo and in the colonic organoids to investigate the cell lineage differentiation-related mechanism of SNS. Furthermore, potential active ingredients from SNS were predicted by network pharmacology analysis. RESULTS: SNS dramatically suppressed DSS-induced acute colonic inflammation in mice. RNA-sequencing analysis revealed downregulation of inflammation and apoptosis-related genes, and upregulation of lipid metabolism and proliferation-related genes, such as Irf7, Pparα, Clspn and Hspa5. Additionally, ISC renewal and intestinal secretory cell lineage commitment were significantly promoted by SNS both in vivo and in vitro in colonic organoids, leading to enhanced mucin expression. Furthermore, potential active ingredients from SNS that mediated inflammation, lipid metabolism, proliferation, apoptosis, stem cells and secretory cells were predicted using a network pharmacology approach. CONCLUSION Our study shed light on the underlying mechanism of SNS in attenuating acute colitis from the perspective of ISC renewal and secretory lineage cell differentiation, suggesting a of novel therapeutic strategy against colitis. Please cite this article as: Cai YJ, Lan JH, Li S, Feng YN, Li FH, Guo MY, et al. Sinisan, a compound Chinese herbal medicine, alleviates acute colitis by facilitating colonic secretory cell lineage commitment and mucin production. J Integr Med. 2025; 23(4): 429-444.
Animals ; Drugs, Chinese Herbal/therapeutic use* ; Mice ; Colon/pathology* ; Mucins/metabolism* ; Mice, Inbred C57BL ; Cell Differentiation/drug effects* ; Male ; Colitis/metabolism* ; Cell Lineage/drug effects* ; Dextran Sulfate ; Stem Cells/drug effects* ; Disease Models, Animal

(+)-Strebloside, a significant bioactive compound isolated from the roots of Streblus asper Lour., demonstrates inhibitory effects against multiple malignancies. However, its specific function and underlying mechanistic pathways in Non-Hodgkin lymphoma (NHL) remain unexplored. This investigation sought to elucidate the role and potential mechanisms of (+)-strebloside-induced NHL cell death. The results demonstrated that (+)-strebloside significantly induced apoptosis and ferroptosis in NHL cells, including those from Raji cell-derived xenograft models. Mechanistic analyses revealed that (+)-strebloside enhanced six-transmembrane epithelial antigen of prostate 3 (STEAP3)-induced ferroptosis in NHL, and STEAP3 inhibition reduced the proliferation-inhibitory effects of (+)-strebloside. Furthermore, (+)-strebloside suppressed NHL proliferation through the mitogen-activated protein kinase (MAPK) pathway, and extracellular signal-regulated kinase (ERK) inhibition diminished the proliferation-inhibitory activity induced by (+)-strebloside. These findings indicate that (+)-strebloside presents promising therapeutic potential for NHL treatment.
Humans ; Ferroptosis/drug effects* ; Lymphoma, Non-Hodgkin/physiopathology* ; Cell Line, Tumor ; MAP Kinase Signaling System/drug effects* ; Animals ; Cell Proliferation/drug effects* ; Mice ; Apoptosis/drug effects* ; Membrane Proteins/genetics* ; Xenograft Model Antitumor Assays ; Male ; Mice, Nude

AIM:To investigate the role and mechanisms of p21 activated kinase 1(Pak1)in myocardial met-abolic reprogramming(MR)in diabetes mellitus(DM)mice and its protective effects on cardiomyocytes.METHODS:Pak1flox/flox(Pak1f/f)mice and cardiomyocyte-specific Pak1 knockout(Pak1CKO)mice were randomly divided into 4 groups(n=6 per group):Pak1f/f group,Pak1f/f+DM group,Pak1CKO group,and Pak1CKO+DM group.Diabetes was induced by in-traperitoneal injection of streptozotocin.Cardiac function was evaluated by echocardiography 6 weeks after successful mod-eling.Ventricular myocardium was harvested after euthanasia.Myocardial pathological changes and lipid accumulation were assessed by HE staining and oil red O staining.Protein expression levels of Pak1,p-Pak1,peroxisome proliferator-activated receptor α(PPARα),PPARγ,lipoprotein lipase(LPL),carnitine palmitoyltransferase 1(CPT1),scavenger receptor B2(SCARB2/CD36),fatty acid binding protein 3(FABP3),pyruvate dehydrogenase kinase 4(PDK4)and the ratio of phosphorylated pyruvate dehydrogenase E1α subunit(p-PDHA1)to PDHA1 were determined by Western blot.The mRNA expression levels of diacylglycerol acyltransferase 1/2(DGAT1/2)were detected by RT-qPCR.RESULTS:Compared with Pak1f/f group,both Pak1f/f+DM and Pak1CKO groups showed significantly decreased left ventricular ejection fraction(EF)and fractional shortening(FS),as well as increased left ventricular end-diastolic volume(LV Vold)and end-systolic volume(LV Vols)(P<0.01).Significant myocardial pathological damage with excessive lipid accumulation was observed.Myocardial p-Pak1 and Pak1 protein expression decreased(P<0.01),while PPARα,PPARγ,CPT1,LPL,CD36,FABP3,and PDK4 protein expression significantly increased(P<0.01),along with elevated p-PDHA1/PDHA1 ratio(P<0.01).mRNA expression levels of DGAT1 and DGAT2 were significantly reduced(P<0.01).The Pak1CKO+DM group showed the same trend as the Pak1f/f+DM group but with greater severity;compared with the Pak1f/f+DM group,the Pak1CKO+DM group still showed significant differences in all indicators except EF,FS,and LVVold(P<0.05 or P<0.01).CONCLUSION:Myocardial Pak1 protein expression is suppressed in diabetic mice,which mediates MR through activation of the PPARα/γ signaling pathway,resulting in increased myocardial fatty acid uptake and utiliza-tion while reducing glucose oxidation capacity.This leads to lipid accumulation in cardiomyocytes,myocardial damage,and cardiac systolic and diastolic dysfunction.

Objective:To investigate the diagnostic and surgical strategies in managing patients with coexisting thyroid carcinoma and cervical vagal schwannoma.Methods:We retrospectively analyzed three cases treated at Guangzhou First People’s Hospital between Jun. 2019 and Dec. 2024.Results:All patients presented with neck masses or thyroid nodules. Ultrasonography identified suspicious malignant thyroid nodules and lateral neck lesions—interpreted as metastatic lymphadenopathy in two cases and as a possible nerve sheath tumor in one. Fine-needle aspiration cytology (FNAC) of thyroid nodules yielded Bethesda VI results in all cases. FNAC of the lateral neck lesions revealed no evidence of malignancy in all cases; in one case, a core needle biopsy confirmed a vagal schwannoma. Two patients underwent MRI, which supported the diagnosis of vagal schwannoma. Surgical management included single-stage resection in two patients and staged surgery in one. Postoperative histopathology confirmed papillary thyroid carcinoma and schwannoma in all cases. Transient postoperative hoarseness occurred in two patients, with recovery within 3-6 months.Conclusions:When thyroid carcinoma is complicated by cervical vagal schwannoma, the schwannoma may be misdiagnosed as metastatic cervical lymph nodes, potentially leading to iatrogenic vagus nerve injury during neck dissection. Although cervical vagal schwannoma can be diagnosed preoperatively, simultaneous surgical treatment of both conditions may increase the risk of vocal cord paralysis, and in severe cases, result in airway compromise. Therefore, comprehensive preoperative evaluation is important. It is recommended to classify cases based on the anatomical locations of the thyroid carcinoma and vagal schwannoma to guide surgical planning, and to use intraoperative nerve monitoring to enhance surgical safety and avoid serious complications.

Objective:To investigate the diagnostic and surgical strategies in managing patients with coexisting thyroid carcinoma and cervical vagal schwannoma.Methods:We retrospectively analyzed three cases treated at Guangzhou First People’s Hospital between Jun. 2019 and Dec. 2024.Results:All patients presented with neck masses or thyroid nodules. Ultrasonography identified suspicious malignant thyroid nodules and lateral neck lesions—interpreted as metastatic lymphadenopathy in two cases and as a possible nerve sheath tumor in one. Fine-needle aspiration cytology (FNAC) of thyroid nodules yielded Bethesda VI results in all cases. FNAC of the lateral neck lesions revealed no evidence of malignancy in all cases; in one case, a core needle biopsy confirmed a vagal schwannoma. Two patients underwent MRI, which supported the diagnosis of vagal schwannoma. Surgical management included single-stage resection in two patients and staged surgery in one. Postoperative histopathology confirmed papillary thyroid carcinoma and schwannoma in all cases. Transient postoperative hoarseness occurred in two patients, with recovery within 3-6 months.Conclusions:When thyroid carcinoma is complicated by cervical vagal schwannoma, the schwannoma may be misdiagnosed as metastatic cervical lymph nodes, potentially leading to iatrogenic vagus nerve injury during neck dissection. Although cervical vagal schwannoma can be diagnosed preoperatively, simultaneous surgical treatment of both conditions may increase the risk of vocal cord paralysis, and in severe cases, result in airway compromise. Therefore, comprehensive preoperative evaluation is important. It is recommended to classify cases based on the anatomical locations of the thyroid carcinoma and vagal schwannoma to guide surgical planning, and to use intraoperative nerve monitoring to enhance surgical safety and avoid serious complications.

[Objective]To explore the incremental cost of central line-associated bloodstream infections(CLABSI)after central venous catheterization(CVC)in critically ill patients in the intensive care unit(ICU),as well as the main cost of nosocomial infection prevention and control.By comparing these two costs,the medical personnel to pay more attention should CLABSI prevention and control from the perspectives of medical quality and economic benefits,and promote the implementation of prevention and control measures.[Methods]Cluster sampling was used to select 126 critically ill patients who underwent CVC in the ICU of a tertiary traditional Chinese medicine hospital from January 2021 to December 2023,including 65 cases in the CLABSI group and 61 in the non-CLABSI group.Patients'data were retrospectively collected from the hospital medical records,including the disease type,gender,age,length of hospital stay,outcome,and hospitalization expenses.The costs of different hand hygiene methods and differing approaches to environmental cleaning and disinfection were analyzed and compared.[Results]There were significant differences in the length of hospital stay(Z=-5.35,P<0.05)and total hospitalization expenses(Z=-6.79,P<0.05)between the CLABSI and non-CLABSI group.Total hospitalization expenses showed significant differences among patients with different lengths of hospital stay(H=43.01,P<0.05),with much higher median one in those with 60 or more days of hospital stay than other patients.Greater differences of median total hospitalization expenses were found in males than in females(Z=-3.98,P<0.05),as well as in patients aged 60-80 years than in patients of other ages(Z=-5.79,P<0.05).[Conclusions]The occurrence of CLABSI significantly increases the ICU patients'length of hospital stay and hospitalization expenses.There are differences in the costs of different hand hygiene methods and differing approaches to environmental cleaning and disinfection,but these costs are acceptable compared to the incremental costs directly attributable to CLABSI.Therefore,medical institutions should attach importance to the investment in prevention and control of nosocomial infections such as hand hygiene and environmental cleaning and disinfection,formulate practical,reasonable and feasible plans,and ensure their implementation,in order to avoid nosocomial infections,improve the medical quality,effectively control patients'length of hospital stay and hospitalization costs,and strive to maintain patient safety.

BACKGROUND:Gait movements are one of the important characteristics exhibited by athletes during exercise,reflecting their physical condition and athletic ability.In a state of fatigue,athletes may exhibit abnormal gait movements,such as reduced stride and body shaking,which can cause harm to their bodies.OBJECTIVE:To promote technological progress in the field of sports science by applying advanced algorithms and data analysis techniques to the training practice of athletes,so as to further improve the recognition accuracy of gait movements under sports fatigue.METHODS:A gait recognition method for athletes in fatigue state was based on fish swarm algorithm.By utilizing the normalized autocorrelation function and the principle of motion energy distribution,a single cycle gait energy map of athletes was obtained.Singular value decomposition was used to transform the image to highlight visual differences,generating a gait energy map of athletes.A convolutional neural network was used to construct a gait action recognition model,and the parameters of the model were solved using the fish swarm algorithm to improve the accuracy and efficiency of fatigue gait action recognition.RESULTS AND CONCLUSION:(1)The fish swarm algorithm had a small loss value in gait action recognition,and could accurately and quickly identify the gait actions of athletes,and dynamically monitor their physical fatigue.(2)The research on fatigue gait recognition of athletes based on fish swarm algorithm could effectively identify the gait movements of athletes in fatigue state and achieve accurate capture of subtle gait changes.(3)The system stability of this method is good,which can reduce the volatility of experimental test results and improve recognition efficiency,can more effectively manage sports fatigue and prevent sports injuries.In addition,when the gait characteristics of normal people change significantly,the system can give an early warning,indicating that the individual may be in a state of fatigue and need to rest or adjust the intensity of activity.