ObjectiveTo investigate the effects of Huanglian Jiedutang （HLJDT） on cognitive function in mice with ischemic stroke （IS） and to elucidate whether its neuroprotective effects are mediated by inhibition of the nuclear factor-κB （NF-κB） signaling pathway and subsequent suppression of NF-κB-regulated neuronal apoptosis. MethodsAn IS model was established using middle cerebral artery occlusion （MCAO）. Sixty C57BL/6J mice were randomly assigned to five groups （n =12 per group）， i.e.， sham operation， model， HLJDT low-dose （3.9 g·kg^-1·d^-1）， HLJDT high-dose （7.8 g·kg^-1·d^-1）， and Ginkgo biloba extract （GBE， 31.2 mg·kg^-1·d^-1）. Post-operatively， neurological deficit scores （Longa score）， cerebral infarct volume assessed by 2，3，5-triphenyltetrazolium chloride （TTC） staining， and brain water content were evaluated. Learning and memory were assessed using new object recognition （NOR） and fear conditioning （FC） tests. Hippocampal pathology was examined via hematoxylin and eosin （HE） staining. Immunofluorescence detected expression of glial fibrillary acidic protein （GFAP， astrocyte marker）， cellular oncogene Fos （c-Fos， neuronal activation marker）， and glutamate decarboxylase 65 （GAD65）. Western blot measured nuclear factor-κB inhibitor protein α （IκBα）， phosphorylated IκBα （p-IκBα）， NF-κB p65， phosphorylated NF-κB p65 （p-NF-κB p65）， ionic calcium binding adapter molecule 1 （Iba-1）， tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and apoptosis-related proteins， such as cleaved cysteinyl aspartate-specific protease 3 （Caspase-3）， B-cell lymphoma 2 （Bcl-2）， and Bcl-2-associated X protein （Bax）. Real-time quantitative PCR （Real-time PCR） was used to assess mRNA levels of Iba-1， TNF-α， IL-1β， NF-κB p65， cleaved Caspase-3， Bax， and Bcl-2. ResultsCompared with the sham group， the model group exhibited significantly increased neurological deficit scores， brain water content， and cerebral infarct volume （P<0.01）. Hippocampal CA1 neurons were disorganized， showing nuclear pyknosis and karyolysis. NOR exploration time and FC freezing time were significantly reduced （P<0.01）. GFAP and c-Fos expression were increased， while GAD65 expression was decreased （P<0.01）. Cleaved Caspase-3 and Bax were upregulated， Bcl-2 was downregulated， and the Bax/Bcl-2 ratio was elevated （P<0.01）. Expression levels of p-IκBα， p-NF-κB p65， IL-1β， TNF-α， and Iba-1 were significantly increased （P<0.01）. Compared with the model group， HLJDT high-dose， low-dose， and GBE groups showed significant improvements in all parameters （P<0.01）. Among them， the HLJDT high-dose group showed the most pronounced neuronal structural recovery and superior performance in NOR and FC tests （P<0.01）. In this group， GFAP and c-Fos decreased， GAD65 increased （P<0.01）， apoptosis-related protein expression was reversed， and NF-κB signaling and related inflammatory factor expression were suppressed （P<0.01）. ConclusionHLJDT ameliorates cognitive dysfunction in mice after IS， potentially by inhibiting the NF-κB signaling pathway， thereby reducing neuroinflammation and hippocampal neuronal apoptosis.

ObjectiveBy establishing a light sleep deprivation （SD） model and using the traditional Chinese medicine （TCM） compound Sancao Anshen prescription for intervention， this study aims to observe one of the effects of Sancao Anshen prescription on the sleep and depressive states of zebrafish in the SD model and explore the mechanism of action of this drug in regulating the cyclic adenosine monophosphate （cAMP）/protein kinase A （PKA） signaling pathway. MethodsA total of 240 six-month-old wild-type AB zebrafish were randomly divided into a blank group， a model group， low-dose， medium-dose， and high-dose groups of Sancao Anshen prescription （0.28， 0.83， 2.48 g·L^-1）， and a melatonin group （0.2 g·L^-1）， with 40 fish in each group. Except for the blank group， all others were exposed to LED lights （150 lux） for three days to construct the sleep deprivation model， and were treated with the corresponding doses of Sancao Anshen decoction and melatonin solution for three days. 24 h movement behavior was used to detect diurnal movement trajectories. A T-shaped maze was employed to detect learning and memory functions， and a new tank experiment was conducted to detect depression-like behaviors in zebrafish. Hematoxylin-eosin （HE） staining was used to observe the morphology of hypothalamic neurons， and transmission electron microscopy was utilized to observe the ultrastructure of hypothalamic cells. Immunohistochemical （IHC） was used to measure the positive expression of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in the hypothalamus， and enzyme-linked immunosorbent assay （ELISA） was employed to determine the content of cAMP， 5-hydroxytryptamine （5-HT）， gamma-aminobutyric acid （GABA）， and glutamic acid （Glu） in brain tissue. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to measure the expression of PKA， cAMP response element-binding （CREB）， and brain-derived neurotrophic factor （BDNF） mRNAs and proteins and their phosphorylation levels （p-PKA， p-CREB） in brain tissue. ResultsCompared with those in the blank group， the resting time and resting count in the model group decreased significantly （P<0.01）， while the gross motor time and gross motor count increased significantly （P<0.01）. The latency time to enter the EC region in each administration group increased significantly （P<0.01）. The exploration time towards the top and the number of times entering the top decreased significantly （P<0.01）， and the incubation period of the first ascent increased significantly （P<0.01）. The number of hypothalamic neurons decreased significantly， and the neurons exhibited irregular shapes， sparse arrangement， and nuclear condensation. Nuclear collapse， nuclear membrane rupture and dissolution， chromatin condensation， mitochondrial swelling and deformity， and plate-like cristae rupture or disappearance were observed. The positive expression of TNF-α and IL-1β in the hypothalamus was significantly decreased （P<0.01）. The content of cAMP， GABA， and 5-HT in brain tissue was significantly downregulated， while the content of Glu was significantly upregulated （P<0.01）. The mRNA of PKA， CREB， and BDNF was significantly downregulated （P<0.01）， and the protein expression of p-PKA， p-PKA/PKA， p-CREB， p-CREB/CREB， and BDNF was significantly decreased （P<0.05， P<0.01）. Compared with those in the model group， the resting time in the medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group increased （P<0.05， P<0.01）， and the resting count in each administration group significantly increased （P<0.05， P<0.01）. The gross motor time and gross motor count significantly decreased （P<0.05， P<0.01）. The latency time to enter the EC region decreased （P<0.01）， and the exploration time towards the top increased （P<0.01）. The time for the first ascent in the high-dose group of Sancao Anshen prescription and the melatonin group was shortened （P<0.05， P<0.01）， and the number of times entering the top increased （P<0.01）. The morphology of neurons in each administration group improved， with the gap decreased， the nuclear membrane relatively intact， and mitochondrial swelling improved. The positive expression of IL-1β in each administration group significantly decreased （P<0.01）. The positive expression of TNF-α in the medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group significantly decreased （P<0.01）. The Glu content in the low-dose group of Sancao Anshen prescription decreased， while cAMP and GABA levels increased （P<0.05， P<0.01）. There was no statistically significant difference in 5-HT content. The medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group all showed significant increases in cAMP， 5-HT， and GABA levels （P<0.01）， and a significant decrease in Glu content （P<0.01）. The mRNA of CREB in the low-dose group of Sancao Anshen prescription was significantly upregulated （P<0.01）， while there was no statistically significant difference in the mRNA of PKA and BDNF. The mRNA of PKA， CREB， and BDNF in the medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group were all significantly upregulated （P<0.01）. The protein expression of p-PKA， p-PKA/PKA， p-CREB， p-CREB/CREB， and BDNF in the high-dose group of Sancao Anshen prescription and the melatonin group increased significantly （P<0.05， P<0.01）， and the protein expression of p-PKA， p-CREB， and BDNF in the medium-dose group of Sancao Anshen prescription increased （P<0.05）. ConclusionThe improvement of sleep and depressive states in zebrafish with the SD model by Sancao Anshen prescription may be related to the inhibition of inflammatory factors such as TNF-α and IL-1β， the reduction in Glu， and the elevation in the content of neurotransmitters such as GABA and 5-HT via the cAMP/PKA signaling pathway.

Objective: To evaluate the diagnostic performance of magnetic resonance spectroscopy (MRS) in discriminating fumarate hydratase-deficient (FH-d) uterine leiomyomas (ULs) from FH-preserved ULs. Materials and Methods: This study consisted of three stages, with independent cohorts recruited for each stage: 1) sample-size estimation was retrospectively performed on UL specimens (diameter ≥3 cm; age, 20–40 years) from our database with immunohistochemistry (IHC) for 2-succinocysteine (2-SC) as the reference, without genetic testing, 2) MRS sequence optimization in confirmed FH germline mutation participants with ultrasound-detected ULs (diameter ≥3 cm), without IHC analysis, and 3) prospective diagnostic test accuracy was evaluated in consecutive participants with ultrasound-detected ULs (diameter ≥3 cm;age, 20–40 years), using IHC for 2-SC for determining the FH status and subsequent genetic testing in those with positive 2-SC results to identify whether FH mutations were germline or somatic in origin. The choline and fumarate peaks in MRS were classified as positive, negative, or technical failure (TF). TFs were analyzed separately and excluded from the primary diagnostic accuracy calculations. T1-, T2-, and diffusion-weighted images were interpreted as hyperintense or hypointense. The enhancement rate and apparent diffusion coefficient were also acquired. Diagnostic performance was compared between MRS and various magnetic resonance imaging (MRI) features. Results: The optimal MRS parameters for the fumarate peak were echo time (TE) = 140 ms and an average of 256. Among the 360 prospective participants, 37 were confirmed to have FH-dULs. MRS showed positive fumarate peaks in 35 of 37 FH-dULs.After excluding six TFs, the positive fumarate peak on MRS showed 94.6% (35/37) sensitivity, 99.7% (316/317) specificity, and 99.2% (351/354) accuracy, all of which were significantly superior to those of other MRI features (P ≤ 0.002). Conclusion A positive fumarate peak on MRS may be a useful imaging biomarker for diagnosing FH-dULs.

Objective: To assess the association of family food environment with dietary and nutrition literacy among junior and senior high school students, providing a basis for nutritional health education and adolescent nutritional improvement. Methods: From January 9 to February 10, 2025, a total of 1 473 junior and senior high school students aged 12-20 were recruited by using a cluster random sampling method combined with convenient sampling from 15 provinces (municipalities and autonomous regions) across China. Data on basic demographic characteristics, home food environment, and dietary and nutrition literacy were collected by using the Food and Nutritional Literacy Questionnaire for Chinese School age Children. Multiple linear regression and Logistic regression models were employed to analyze the association of the home food environment on adolescent dietary and nutrition literacy. Results: The total score of dietary and nutrition literacy among junior and senior high school students was (65.28±9.03). The score in the cognitive domain (67.59±10.56) was higher than that in the skill domain (60.74±9.15), and the score for food preparation skills (56.60±14.28) was the lowest. Students aged 16-20, senior high school students, those for only children, non boarding students, those with urban household registration, those from wealthier families, those whose primary caregivers were parents with higher education levels, and those who previously received nutrition and health education had significantly higher dietary and nutrition literacy scores compared to their respective counterpart groups ( t/F =-8.32, -8.14, 4.76, -2.70, 7.15, 18.29, 10.68, 11.66 , all P <0.05). Multiple linear regression analysis revealed that family food environment was associated with dietary and nutrition literacy, and the model was statistically significant ( R 2=0.20, F=20.54, P <0.05). Lower availability of fruits at home in the past month and lower frequency of family communication regarding dietary and nutrition were associated with lower dietary nutrition literacy scores among junior and senior high school students ( β =-1.35, -1.67, both P <0.01). Among junior and senior high school students, lower frequency of dining out with the family, watching videos during family meals, and being urged by parents to eat more food in the past month were associated with higher scores on dietary nutrition literacy ( β =1.35, 1.13, 0.71, all P <0.01). Logistic regression model results indicated that watching videos during family meals ( OR =1.70) and family communication regarding dietary and nutrition ( OR =0.51) were significantly associated with dietary and nutrition literacy levels (both P <0.05). Conclusions The score of dietary and nutrition literacy among junior and senior high school students is relatively low, and the family food environment is a key factor. It is recommended that nutrition education and improvement initiatives for junior and senior high school students prioritize attention on children and adolescents from households with less favorable food environments.

Genomic imprinting refers to the phenomenon of differential expression of two alleles due to their different parental origins. Genes that produce genomic imprinting are usually called imprinted genes. The genetic effect caused by the presence of imprinted genes is called parent-of-origin effect. Parent-of-origin effect and genomic imprinting play important roles in the pathophysiological mechanism and occurrence and development of cardio-metabolic diseases. In-depth exploration of the law and potential roles of imprinted genes and parent-of-origin effects will help to better understand the mechanism of cardio-metabolic diseases, and also provide important theoretical basis for the precise treatment of diseases related to imprinted genes.

Objective:Chimeric antigen receptor T-cell(CAR-T)immunotherapy has made major breakthroughs in the treatment of blood tu-mors.However,current CAR-T therapies face several limitations:they require autologous cells,involve a lengthy and costly production pro-cess,and use lentiviral transduction that carry risk of insertional carcinogenesis due to random integration.Therefore,there is an urgent need to develop a universal cost-effective cancer immunotherapy method generating CAR-T cells for in vivo cancer immunotherapy.Meth-ods:This study successfully established an exosome-mediated,T-cell targeted delivery system,demonstrating both precise design and func-tional efficacy for biomedical applications.To optimize CAR-T cell generation the transfection dose was adjusted,and the kinetics of CAR-T cell percentage were recorded.The cytotoxicity of the resulting CAR-T cells was evaluated in vitro by calcein-AM release.To test the tumor-killing in vivo of engineered exosomes,human PBMCs were injected into NPG mice via the tail vein to establish humanized mice,followed by intravenous injection of tumor cells to induce cancer.Results:To overcome the limitations of conditional autologous CAR-T cells,we de-veloped a T cell-targeted exosome system capable of specifically targeting human CD3+,CD4+,and CD8+T cells.CAR-T production was dose-dependent,with transfection efficiency reaching upto 97.8%at 106 particles/cell.Both in vitro cytotoxicity assays and in vivo animal experi-ments demonstrated that exosome-incubated CAR-T cells effectively eliminated CD19-positive Raji cells,highlighting their specificity and therapeutic potential in antigen-directed applications.Conclusions:We successfully established a CD8-targeting exosome delivery system for CAR-T cell production capable of transforming CD8+T cells into functional CAR-T cells,which showed significant tumor-killing ability in vitro and in mice.Compared with the traditional lentiviral vector for the preparation of CAR-T cells in vitro,in vivo-reprogrammed CAR-T cells us-ing our CD8-targeted exosome delivery system,with higher transfection efficiency,shorter production period,lower cost,and eliminated the risk of insertion carcinogenesis.This strategy promises to bring a new era of universal CAR-T medicine,which can improve cancer immuno-therapy and may hold promise as a therapeutic platform to treat various diseases.

Klebsiella pneumoniae,as a clinically prevalent opportunistic pathogen,ranks as the second most com-monly detected pathogen in clinical isolates in China.The extensive clinical use of carbapenem antibiotics has led to a high global detection rate of carbapenem-resistant K.pneumoniae(CRKP).Characterized by complex resist-ance mechanisms and diverse evolutionary pathways,CRKP infections pose significant challenges in prevention and treatment,with high associated mortality rates,creating substantial obstacles for clinical anti-infective therapy.In recent years,the emergence and global spread of carbapenem-resistant hypervirulent K.pneumoniae(CR-hvKP)have escalated into a major public health threat.Notably,hypervirulent K.pneumoniae isolates carry-ing carbapenem resistance genes are rapidly disseminating worldwide,causing fatal infections even in immunocom-petent individuals.This article systematically reviews the latest research advances on the resistance mechanisms,evolutionary pathways,adaptive changes,and clinical management strategies of CR-hvKP,aiming to deepen un-derstanding of this"superbug"and provide a theoretical foundation for clinical prevention and control.

Genomic imprinting refers to the phenomenon of differential expression of two alleles due to their different parental origins. Genes that produce genomic imprinting are usually called imprinted genes. The genetic effect caused by the presence of imprinted genes is called parent-of-origin effect. Parent-of-origin effect and genomic imprinting play important roles in the pathophysiological mechanism and occurrence and development of cardio-metabolic diseases. In-depth exploration of the law and potential roles of imprinted genes and parent-of-origin effects will help to better understand the mechanism of cardio-metabolic diseases, and also provide important theoretical basis for the precise treatment of diseases related to imprinted genes.

Objective:To evaluate the application potential of the bimodal ultrasound/near-infrared (NIR) composite nanoscale probe Arg-Gly-Asp (RGD)/Ag 2Te/perfluoropentane (PFP) @ mesoporous silica nanoparticles (MSN) in the diagnosis and treatment of microvascular diseases. Methods:Nanoprobes loaded with RGD, PFP and Ag 2Te were prepared by ultrasound sonication and carbodiimide method. The characterization of the nanoprobes was determined. The imaging performance, photothermal response, and target-seeking ability of the nanoprobes under NIR irradiation were verified. The biosafety of the nanoprobes was examined, and the thrombolytic ability of the nanoprobes was evaluated. The mice were observed to visualize microvessels of the abdominal wall under the NIR-Ⅱ imaging, and the microvascular visualization ability of the nanoprobes was evaluated. Results:The particle size of nanoprobes was (205.3±2.9) nm and the potential was (2.05±0.58) mV. The coupling rate of the RGD was (82.27±0.36)%, the encapsulation rate of the quantum dots was (80.80±3.26)%, and the photostability of the quantum dots was good. The fluorescence intensity was enhanced with the increase of the mass concentration of RGD/Ag 2Te/PFP@MSN, and the warming effect was more obvious. After ultrasound and NIR irradiation, the thrombolysis rate was significantly increased. RGD/Ag 2Te/PFP@MSN successfully realized NIR-Ⅱ fluorescence imaging of mice microvessels. The cytotoxicity assay and hemolysis assay showed that the probe had a good biosafety. Conclusion:The RGD/Ag 2Te/PFP@MSN nanoprobe is a potential strategy for targeted therapy of thrombotic diseases, combining dual-modality therapy of ultrasound and NIR to offer new possibilities for non-invasive and visual diagnosis and treatment of microvascular embolism.

Objective:Chimeric antigen receptor T-cell(CAR-T)immunotherapy has made major breakthroughs in the treatment of blood tu-mors.However,current CAR-T therapies face several limitations:they require autologous cells,involve a lengthy and costly production pro-cess,and use lentiviral transduction that carry risk of insertional carcinogenesis due to random integration.Therefore,there is an urgent need to develop a universal cost-effective cancer immunotherapy method generating CAR-T cells for in vivo cancer immunotherapy.Meth-ods:This study successfully established an exosome-mediated,T-cell targeted delivery system,demonstrating both precise design and func-tional efficacy for biomedical applications.To optimize CAR-T cell generation the transfection dose was adjusted,and the kinetics of CAR-T cell percentage were recorded.The cytotoxicity of the resulting CAR-T cells was evaluated in vitro by calcein-AM release.To test the tumor-killing in vivo of engineered exosomes,human PBMCs were injected into NPG mice via the tail vein to establish humanized mice,followed by intravenous injection of tumor cells to induce cancer.Results:To overcome the limitations of conditional autologous CAR-T cells,we de-veloped a T cell-targeted exosome system capable of specifically targeting human CD3+,CD4+,and CD8+T cells.CAR-T production was dose-dependent,with transfection efficiency reaching upto 97.8%at 106 particles/cell.Both in vitro cytotoxicity assays and in vivo animal experi-ments demonstrated that exosome-incubated CAR-T cells effectively eliminated CD19-positive Raji cells,highlighting their specificity and therapeutic potential in antigen-directed applications.Conclusions:We successfully established a CD8-targeting exosome delivery system for CAR-T cell production capable of transforming CD8+T cells into functional CAR-T cells,which showed significant tumor-killing ability in vitro and in mice.Compared with the traditional lentiviral vector for the preparation of CAR-T cells in vitro,in vivo-reprogrammed CAR-T cells us-ing our CD8-targeted exosome delivery system,with higher transfection efficiency,shorter production period,lower cost,and eliminated the risk of insertion carcinogenesis.This strategy promises to bring a new era of universal CAR-T medicine,which can improve cancer immuno-therapy and may hold promise as a therapeutic platform to treat various diseases.