ObjectivePost-ischemic acute inflammation and the subsequent persistent dysregulation of the immune microenvironment represent major pathological drivers that aggravate neuronal injury and severely restrict functional recovery following ischemic stroke. Although current reperfusion therapies partially restore blood flow, they fail to effectively modulate the secondary inflammatory cascade and oxidative stress, which remain critical barriers to neurological restoration. To address this challenge, this study aimed to engineer and systematically evaluate a biomimetic nanosystem composed of transforming growth factor-β1 (TGF-β1)-loaded platelet membrane-camouflaged lipid nanoparticles (PLP). This nanosystem was designed to achieve dual lesion-targeted delivery and immune microenvironment remodeling. By verifying its spatiotemporal accumulation, anti-inflammatory activity, and neuroprotective efficacy, we sought to establish an integrated therapeutic strategy that simultaneously enables lesion targeting, immune regulation, and functional recovery after ischemic injury. MethodsThe physicochemical properties of PLP, including hydrodynamic particle size, zeta potential, structural stability, and morphology, were characterized using dynamic light scattering, zeta potential analysis, and transmission electron microscopy. The preservation of platelet membrane-derived adhesion and immunoregulatory proteins was confirmed by SDS-PAGE through comparative analysis of protein band profiles between PLP and native platelet membranes. The in vitro biological activities of PLP were evaluated using two complementary cellular models. LPS-induced M1-polarized RAW264.7 macrophages were employed to assess inflammatory modulation, while oxygen glucose deprivation/reperfusion (OGD/R)-induced BV2 microglial cells and SH-SY5Y neuronal cells were utilized to investigate neuroinflammatory regulation and neuronal protection. For in vivo validation, a transient middle cerebral artery occlusion (tMCAO) mouse model was established to mimic ischemia-reperfusion injury. The spatiotemporal biodistribution and lesion-targeting capability of the PLP were monitored through live fluorescence imaging. Therapeutic efficacy was comprehensively evaluated by triphenyltetrazolium chloride (TTC) staining, glial fibrillary acidic protein (GFAP) immunofluorescence analysis, body weight monitoring, and neurological severity score (NSS) assessment. ResultsPLP nanoparticles displayed a uniform spherical morphology, nanoscale particle size distribution, and stable negative surface charge, indicating favorable colloidal stability and circulation potential. SDS-PAGE results confirmed the effective retention of key platelet membrane proteins associated with endothelial adhesion, immune evasion, and inflammatory regulation, demonstrating the successful biomimetic construction. Optimal therapeutic concentrations were determined in OGD/R-induced BV2 cells, where PLP exhibited excellent cytocompatibility and anti-inflammatory activity.In vitro experiments demonstrated that PLP significantly inhibited the polarization of RAW264.7 macrophages toward the pro-inflammatory M1 phenotype and markedly reduced neuronal apoptosis under ischemia-reperfusion conditions. In vivo fluorescence imaging revealed that PLP rapidly accumulated in the ischemic brain hemisphere and maintained prolonged retention for up to 7 d, suggesting enhanced lesion-specific targeting and sustained drug release. Compared with control group, PLP treatment significantly reduced cerebral infarct volume, attenuated reactive astrogliosis, improved weight recovery, and accelerated neurological functional restoration, as reflected by significantly improved NSS scores. ConclusionThis study establishes a multifunctional biomimetic nanoplatform that integrates platelet membrane-mediated active targeting with the anti-inflammatory, antioxidative, and neuroprotective properties of TGF-β1. The PLP system enables rapid lesion homing and long-term retention while synergistically regulating the post-stroke inflammatory microenvironment by suppressing pro-inflammatory immune activation, reducing neuronal apoptosis, and limiting excessive astrocyte reactivity. Importantly, this study proposes a conceptually therapeutic paradigm that combines targeted delivery with immune microenvironment remodeling to achieve comprehensive neurovascular protection. These findings provide strong experimental evidence supporting the translational potential of biomimetic nanotherapeutics as next-generation precision interventions for ischemic stroke.

AIM:To investigate the therapeutic ef-fect of crocetin on diabetic lower limb ischemia and explore its underlying mechanism.METHODS:Male C57BL/6J mice aged 8 weeks were used to es-tablish a diabetic mouse model through intraperito-neal injection of streptozotocin(STZ).Following successful induction of diabetes,femoral artery li-gation(FAL)surgery was performed to create a model of diabetic lower limb ischemia.Fourteen days after STZ injection,the mice were treated with crocetin(3 mg/kg and 6 mg/kg)by gavage for 21 consecutive days.Post-FAL surgery,Doppler flowmetry was employed to assess blood flow in the mice of each group.Immunofluorescence stain-ing techniques were utilized to observe the expres-sion levels of platelet-endothelial cell adhesion molecule(CD31),α-smooth muscle actin(α-SMA),and inflammatory-related factors including tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),in-terleukin-6(IL-6),and transforming growth factor-β(TGF-β)in the gastrocnemius muscle of diabetic mice with lower limb ischemia.RESULTS:Com-pared to the normal group,the model group exhib-ited significantly elevated blood glucose levels and significantly reduced lower limb blood flow(P＜0.05).While CD31 and α-SMA expression showed no significant change,the expression levels of TNF-α,IL-1β,IL-6,and TGF-β were significantly in-creased(P＜0.05,P＜0.01,P＜0.05,P＜0.05).Com-pared to the model group,crocetin-treated groups showed no significant change in blood glucose lev-els but demonstrated significantly increased lower limb blood flow(P＜0.05).The high-dose crocetin group(6 mg/kg)significantly enhanced CD31 andα-SMA expression(P＜0.0001,P＜0.05)and signifi-cantly reduced the expression levels of IL-1β,IL-6,and TGF-β(P＜0.001,P＜0.05,P＜0.05).CONCLU-SION:Crocetin may exert its beneficial effects on diabetic lower limb ischemia through anti-inflam-matory and angiogenic mechanisms.

Background and Aims:Primary hepatic diffuse large B-cell lymphoma(DLBCL)is rare and carries a poor prognosis,with no large-scale evidence supporting the value of chemotherapy.This study used real-world,large-sample data from the SEER database to evaluate the prognostic impact of chemotherapy in patients with primary hepatic DLBCL.Methods:Clinical data of patients pathologically diagnosed with primary hepatic DLBCL between 2000 and 2019 were extracted from the SEER database and grouped according to whether they received chemotherapy.Kaplan-Meier survival analysis,Cox proportional hazards models,and random survival forest models were employed to identify factors influencing overall survival(OS)and cancer-specific survival(CSS),and subgroup analyses were performed.Results:A total of 594 patients were included,with a median age of 66 years;435(73.2%)received chemotherapy.After a median follow-up of 17.5 months,the median OS was 81(7-173)months in the chemotherapy group versus 11(2-171)months in the non-chemotherapy group.The OS and CSS rates in the chemotherapy group were significantly higher than those in the non-chemotherapy group(67.99%vs.47.77%;71.03%vs.52.87%,both P<0.05).Multivariate analysis showed that chemotherapy was an independent protective factor for OS(HR=0.39,95%CI=0.31-0.48)and CSS(HR=0.37,95%CI=0.29-0.48).In the random survival forest model,chemotherapy ranked highest in variable importance.Subgroup analyses produced consistent results.Conclusion:Chemotherapy is an independent prognostic factor for patients with primary hepatic DLBCL and can significantly improve survival.The R-CHOP regimen may be the preferred therapeutic option.

Background and Aims:Primary hepatic diffuse large B-cell lymphoma(DLBCL)is rare and carries a poor prognosis,with no large-scale evidence supporting the value of chemotherapy.This study used real-world,large-sample data from the SEER database to evaluate the prognostic impact of chemotherapy in patients with primary hepatic DLBCL.Methods:Clinical data of patients pathologically diagnosed with primary hepatic DLBCL between 2000 and 2019 were extracted from the SEER database and grouped according to whether they received chemotherapy.Kaplan-Meier survival analysis,Cox proportional hazards models,and random survival forest models were employed to identify factors influencing overall survival(OS)and cancer-specific survival(CSS),and subgroup analyses were performed.Results:A total of 594 patients were included,with a median age of 66 years;435(73.2%)received chemotherapy.After a median follow-up of 17.5 months,the median OS was 81(7-173)months in the chemotherapy group versus 11(2-171)months in the non-chemotherapy group.The OS and CSS rates in the chemotherapy group were significantly higher than those in the non-chemotherapy group(67.99%vs.47.77%;71.03%vs.52.87%,both P<0.05).Multivariate analysis showed that chemotherapy was an independent protective factor for OS(HR=0.39,95%CI=0.31-0.48)and CSS(HR=0.37,95%CI=0.29-0.48).In the random survival forest model,chemotherapy ranked highest in variable importance.Subgroup analyses produced consistent results.Conclusion:Chemotherapy is an independent prognostic factor for patients with primary hepatic DLBCL and can significantly improve survival.The R-CHOP regimen may be the preferred therapeutic option.

AIM:A strain was isolated and identified as the H3N8 subtype of the avian influenza virus from a sick chicken at a farm in Yangjiang,Guangdong Province,named A/chicken/Yangjiang/552/2023(abbreviated as YJ/552).The aim of this research is to determine its genetic evolution,biological properties and pathogenicity in hamsters.This study may provide a theoretical strategy for preventing and treating the H3N8 subtype avian influenza virus-induced epidemic.METHODS:A strain of H3N8 avian influenza virus from chickens was characterised by phylogenetic analy-sis,antigenic diversity,receptor-binding specificity,neuraminidase activity,replication,and transmission in hamsters and a systematic pathological analysis was conducted.RESULTS:This novel avian influenza virus was generated through complex recombination of Eurasian avian H3 genes,North American avian N8 genes and six internal genes of H9N2 sub-type AIV.The cleavage site of the outer protein,HA,was PEKQTR↓GLF,which is characteristic of the low pathogenic avian influenza virus.The HA gene of YJ/552 exhibited the highest nucleotide homology with A/China/ZMD-22-2/2022(H3N8)at 99.09%,while the NA gene showed the highest homology with A/chicken/Dongguan/879/2022(H3N8)at 99.01%.This strain preferentially binds to avian-type receptors and could bind to human-type receptors.This virus could effectively replicate in the trachea and lungs of inoculated and contact hamsters.CONCLUSION:YJ/552 is a recombi-nant H3N8 avian influenza virus replicated in the upper respiratory system and transmitted in hamsters.This study pro-vides data support for the early warning and prevention of H3 subtype avian influenza viruses.

AIM:A strain was isolated and identified as the H3N8 subtype of the avian influenza virus from a sick chicken at a farm in Yangjiang,Guangdong Province,named A/chicken/Yangjiang/552/2023(abbreviated as YJ/552).The aim of this research is to determine its genetic evolution,biological properties and pathogenicity in hamsters.This study may provide a theoretical strategy for preventing and treating the H3N8 subtype avian influenza virus-induced epidemic.METHODS:A strain of H3N8 avian influenza virus from chickens was characterised by phylogenetic analy-sis,antigenic diversity,receptor-binding specificity,neuraminidase activity,replication,and transmission in hamsters and a systematic pathological analysis was conducted.RESULTS:This novel avian influenza virus was generated through complex recombination of Eurasian avian H3 genes,North American avian N8 genes and six internal genes of H9N2 sub-type AIV.The cleavage site of the outer protein,HA,was PEKQTR↓GLF,which is characteristic of the low pathogenic avian influenza virus.The HA gene of YJ/552 exhibited the highest nucleotide homology with A/China/ZMD-22-2/2022(H3N8)at 99.09%,while the NA gene showed the highest homology with A/chicken/Dongguan/879/2022(H3N8)at 99.01%.This strain preferentially binds to avian-type receptors and could bind to human-type receptors.This virus could effectively replicate in the trachea and lungs of inoculated and contact hamsters.CONCLUSION:YJ/552 is a recombi-nant H3N8 avian influenza virus replicated in the upper respiratory system and transmitted in hamsters.This study pro-vides data support for the early warning and prevention of H3 subtype avian influenza viruses.

Objective To investigate the effects of estrogen signaling on T-cell recruitment and polarization in acutely injured skeletal muscle.Methods One hundred C57BL/6 male mice,one hundred and eighty C57BL/6 female mice were selected.Twenty-five female mice were ovariectomized(OVX)and 10 male mice were taken as the sham-operated(sham).Then,cardiotoxin(CTX)induced tibialis anterior(TA)injury for preparing mice myoinjury model.Subcutaneous injection of 17β-estradiol(E2)or estrogen receptor antagonist 4-hydroxytamoxifen(4-OHT)was performed.A total of 140 mice(70 males and 70 females)were divided into four group including:PBS-male,CTX-male,PBS-female,and CTX-female.Serum estradiol(E2)levels were measured by ELISA,and muscle injury models were validated via HE staining.Subsequently,20 male and 20 female mice were selected for immunofluorescence(IF)and Real-time PCR to assess estrogen receptors(ER)expression in injured muscle tissue.Further,10 male and 40 female mice were allocated into five experimental groups,including CTX,CTX+E2,CTX+4-OHT,CTX+OVX,CTX+sham.HE staining and IF were performed to evaluate inflammatory infiltration in the injured muscle.Additionally,50 female mice were divided into CTX and CTX+OVX groups,and IF combined with flow cytometry were used to analyze T-cell phenotypes and muscle fiber regeneration in the injured muscle.Results In vivo,serum E2 and myofiber ERβ increased post-injury in mice of both sexes,significantly higher in females.Compared to the control group,E2 alleviated inflammation,OVX exacerbated inflammation,increased CD4+T-cell infiltration,elevated T helper 1 cell(Th1)response,decreased regulatory T cells(Tregs),impaired regeneration.In vitro,IFN-γ/LPS significantly upregulated ERβ in myotubes.Conclusion Estrogen signaling critically regulates muscle inflammation.Estrogen deficiency(OVX)delays repair of skeletal muscle by promoting Th1 response and suppressing Tregs function.

Objective To investigate the status of population dynamics and distribution changes of Aedes aegypti in Guangdong Province.Methods Continuous monitoring was conducted from May 2018 to July 2024 in Wushi Town and Qishui Town,Leizhou City,Zhanjiang City,Guangdong Province.Additionally,a survey of the distribution of Ae.aegypti along the Leizhou Peninsula coast was carried out.Results The density of Ae.aegypti in Zhanjiang showed a gradual decline from 2018 to 2024.The last detection of adult Ae.aegypti in Wushi Town was in September 2021,and the last larva was found in October 2023.No Ae.aegypti was detected in Qishui Town during surveys from 2021 to 2024.A survey of 18 coastal villages in the Leizhou Peninsula revealed no detections of Ae.aegypti.Conclusions This study provides a basis for understanding the distribution and population density fluctuations of Ae.aegypti,assessing its invasion risk,and scientifically conducting relevant prevention and control efforts.

AIM:To investigate the therapeutic ef-fect of crocetin on diabetic lower limb ischemia and explore its underlying mechanism.METHODS:Male C57BL/6J mice aged 8 weeks were used to es-tablish a diabetic mouse model through intraperito-neal injection of streptozotocin(STZ).Following successful induction of diabetes,femoral artery li-gation(FAL)surgery was performed to create a model of diabetic lower limb ischemia.Fourteen days after STZ injection,the mice were treated with crocetin(3 mg/kg and 6 mg/kg)by gavage for 21 consecutive days.Post-FAL surgery,Doppler flowmetry was employed to assess blood flow in the mice of each group.Immunofluorescence stain-ing techniques were utilized to observe the expres-sion levels of platelet-endothelial cell adhesion molecule(CD31),α-smooth muscle actin(α-SMA),and inflammatory-related factors including tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),in-terleukin-6(IL-6),and transforming growth factor-β(TGF-β)in the gastrocnemius muscle of diabetic mice with lower limb ischemia.RESULTS:Com-pared to the normal group,the model group exhib-ited significantly elevated blood glucose levels and significantly reduced lower limb blood flow(P＜0.05).While CD31 and α-SMA expression showed no significant change,the expression levels of TNF-α,IL-1β,IL-6,and TGF-β were significantly in-creased(P＜0.05,P＜0.01,P＜0.05,P＜0.05).Com-pared to the model group,crocetin-treated groups showed no significant change in blood glucose lev-els but demonstrated significantly increased lower limb blood flow(P＜0.05).The high-dose crocetin group(6 mg/kg)significantly enhanced CD31 andα-SMA expression(P＜0.0001,P＜0.05)and signifi-cantly reduced the expression levels of IL-1β,IL-6,and TGF-β(P＜0.001,P＜0.05,P＜0.05).CONCLU-SION:Crocetin may exert its beneficial effects on diabetic lower limb ischemia through anti-inflam-matory and angiogenic mechanisms.

OBJECTIVE: To compare the therapeutic efficacy of portal and tail vein transplantation of bone marrow-derived mesenchymal stem cells (BMSCs) against cholestatic liver fibrosis in mice. METHODS: BMSCs were isolated and co-cultured with starvation-activated hepatic stellate cells (HSCs). HSC activation markers were identified using immunofluorescence and qRT-PCR. BMSCs were injected into the liver tissues of bile duct ligation (BDL) mice via the tail and portal veins. Histomorphology, liver function, inflammatory cytokines, and the expression of key proteins were all determined in the liver tissues. RESULTS: BMSCs inhibited HSC activation by reducing α-SMA and collagen I expression. Compared to tail vein injection, DIL-labeled BMSCs injected through the portal vein maintained a high homing rate in the liver. Moreover, BMSCs transplanted through the portal vein resulted in greater improvement in liver color, hardness, and gallbladder size than did those transplanted through the tail vein. Furthermore, BMSCs injected by portal vein, but not tail vein, markedly ameliorated liver function, reduced the secretion of inflammatory cytokines, including TNF-α, IL-6, and IL-1β, and decreased α-SMA + hepatic stellate cell (HSC) activation and collagen fiber formation. CONCLUSION The therapeutic effect of BMSCs on cholestatic liver fibrosis in mice via portal vein transplantation was superior to that of tail vein transplantation. This comparative study provides reference information for further BMSC studies focused on clinical cholestatic liver diseases.
Animals ; Mice ; Mesenchymal Stem Cell Transplantation ; Liver Cirrhosis/etiology* ; Male ; Cholestasis/therapy* ; Mice, Inbred C57BL ; Hepatic Stellate Cells ; Mesenchymal Stem Cells