ObjectiveTo investigate the molecular mechanism by which Yifei Xuanfei Jiangzhuo prescription regulates the nuclear factor-κB （NF-κB）/NOD-like receptor protein 3 （NLRP3） signaling pathway to improve neuronal function in vascular dementia （VaD） rats. MethodsA VaD model was established by intermittently clamping the bilateral common carotid arteries （CCA） combined with bilateral vascular occlusion （2-VO）. Eighty-four SD rats were randomly divided into a blank group， sham group， model group， piracetam group （0.2 g·kg^-1）， and low-， medium-， and high-dose Yifei Xuanfei Jiangzhuo prescription groups （6.09， 12.18， and 24.36 g·kg^-1）. Drug administration began on day 7 after surgery， once daily for 28 consecutive days. Behavioral experiments were used to evaluate learning and spatial memory. Hematoxylin-eosin （HE） staining was applied to observe pathological morphological changes in the CA1 region of the hippocampus. Transmission electron microscopy was used to examine the ultrastructure of hippocampal neurons. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） was used to detect neuronal apoptosis in the CA1 region. Immunohistochemistry was performed to determine the positive expression rate of neuronal nuclear antigen （NeuN）. Immunofluorescence single staining was used to assess nuclear expression of NF-κB p65 in brain tissue. Western blot was used to detect the protein expression levels of inhibitor of κB kinase （IKK）， NF-κB p65， NLRP3， Caspase-1， apoptosis-associated speck-like protein （ASC）， and interleukin-1β （IL-1β）. ResultsCompared with the blank group， the model group showed a significant reduction in platform-crossing frequency （P0.01）， aggravated hippocampal injury， a significant increase in neuronal apoptosis （P0.05）， decreased NeuN positivity in the CA1 region （P0.05）， increased nuclear expression of NF-κB p65 （P0.05）， and significantly elevated expression of p-IKK， p-NF-κB p65， NLRP3， cleaved Caspase-1， ASC， and cleaved IL-1β （P0.05）. Compared with the model group， all drug-treated groups improved learning and spatial memory in VaD rats， alleviated hippocampal pathological injury and neuronal apoptosis， and protected neuronal ultrastructure. Yifei Xuanfei Jiangzhuo prescription at doses of 12.18 and 24.36 g·kg^-1 reduced hippocampal expression levels of p-IKK， p-NF-κB p65， NLRP3， Caspase-1， ASC， and cleaved IL-1β in VaD rats （P0.05）， showing dose-dependent inhibition of the NF-κB/NLRP3 signaling pathway. ConclusionYifei Xuanfei Jiangzhuo prescription may exert neuroprotective effects by regulating the NF-κB/NLRP3 signaling pathway， thereby reducing neuroinflammation and inhibiting hippocampal neuronal apoptosis.

ObjectiveTo investigate the molecular mechanism by which Yifei Xuanfei Jiangzhuo prescription regulates the nuclear factor-κB （NF-κB）/NOD-like receptor protein 3 （NLRP3） signaling pathway to improve neuronal function in vascular dementia （VaD） rats. MethodsA VaD model was established by intermittently clamping the bilateral common carotid arteries （CCA） combined with bilateral vascular occlusion （2-VO）. Eighty-four SD rats were randomly divided into a blank group， sham group， model group， piracetam group （0.2 g·kg^-1）， and low-， medium-， and high-dose Yifei Xuanfei Jiangzhuo prescription groups （6.09， 12.18， and 24.36 g·kg^-1）. Drug administration began on day 7 after surgery， once daily for 28 consecutive days. Behavioral experiments were used to evaluate learning and spatial memory. Hematoxylin-eosin （HE） staining was applied to observe pathological morphological changes in the CA1 region of the hippocampus. Transmission electron microscopy was used to examine the ultrastructure of hippocampal neurons. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） was used to detect neuronal apoptosis in the CA1 region. Immunohistochemistry was performed to determine the positive expression rate of neuronal nuclear antigen （NeuN）. Immunofluorescence single staining was used to assess nuclear expression of NF-κB p65 in brain tissue. Western blot was used to detect the protein expression levels of inhibitor of κB kinase （IKK）， NF-κB p65， NLRP3， Caspase-1， apoptosis-associated speck-like protein （ASC）， and interleukin-1β （IL-1β）. ResultsCompared with the blank group， the model group showed a significant reduction in platform-crossing frequency （P0.01）， aggravated hippocampal injury， a significant increase in neuronal apoptosis （P0.05）， decreased NeuN positivity in the CA1 region （P0.05）， increased nuclear expression of NF-κB p65 （P0.05）， and significantly elevated expression of p-IKK， p-NF-κB p65， NLRP3， cleaved Caspase-1， ASC， and cleaved IL-1β （P0.05）. Compared with the model group， all drug-treated groups improved learning and spatial memory in VaD rats， alleviated hippocampal pathological injury and neuronal apoptosis， and protected neuronal ultrastructure. Yifei Xuanfei Jiangzhuo prescription at doses of 12.18 and 24.36 g·kg^-1 reduced hippocampal expression levels of p-IKK， p-NF-κB p65， NLRP3， Caspase-1， ASC， and cleaved IL-1β in VaD rats （P0.05）， showing dose-dependent inhibition of the NF-κB/NLRP3 signaling pathway. ConclusionYifei Xuanfei Jiangzhuo prescription may exert neuroprotective effects by regulating the NF-κB/NLRP3 signaling pathway， thereby reducing neuroinflammation and inhibiting hippocampal neuronal apoptosis.

As the important source of bone cells, osteoblasts are involved in bone formation and repair, and play a key role in maintaining bone balance. If the osteogenic differentiation process in vivo is disrupted, a variety of bone-related diseases may occur. Natural products, which have a wide range of sources, a wide variety of physiological activities, and few toxic side-effects, have been found in recent years to be able to regulate osteoblast differentiation. Based on the sources of natural products, this paper reviews the intervention of natural products from plant, animal and microbial sources on osteogenic differentiation, aiming to provide a theoretical basis for natural products in the treatment of bone diseases.

Macrophage extracellular traps(METs)are extracellular fibrous web-like structures produced by macro-phages.Under physiological conditions,METs capture and kill microorganisms by releasing high concentrations of granular proteins,serving as an innate immune defense mechanism and playing a vital protective role in resisting the progression of inflammatory diseases.Excessive release of METs can also exacerbate the inflammatory response and cause further tissue damage.

Objective : To identify autophagy-related genes involved in pulmonary infection caused by the highly drug-resistant and virulent methicillin-resistant Staphylococcus aureus strain USA300 ( USA300) ，and to explore the underlying molecular mechanisms ， thereby providing potential targets for immunotherapy． Methods: The GSE220943 dataset of a USA300-induced pulmonary infection mouse model was obtained from the GEO database． Differentially expressed genes ( DEGs ) were identified using the DESeq2 package． Autophagy-related genes ( ARGs) were retrieved from the MSigDB and Autophagy databases．Weighted gene co-expression network analysis ( WGCNA) was performed to construct gene co-expression modules．Genes overlapping among DEGs，ARGs，and WGCNA modules were identified as autophagy-related DEGs．Gene Ontology ( GO) enrichment analysis was con- ducted using the clusterProfiler R package，while Kyoto Encyclopedia of Genes and Genomes ( KEGG) pathway en- richment analysis was performed via the Metascape platform．Immune cell infiltration was analyzed using the Immu- CellAI-mouse website．A protein - protein interaction ( PPI) network was constructed using the STRING database， and hub genes were identified through topological analysis in Cytoscape． Receiver operating characteristic curve ( ROC) curves were plotted via the website https: / /www．bioinformatics．com．cn． Finally，key gene expression was validated in mouse lung tissues by real-time quantitative reverse transcription PCR ( RT-qPCR) ． Results: A total of 6 135，4 075，3 680，and 2 342 differentially expressed genes ( DEGs) were identified at 12，24，48，and 96 hours post-infection，respectively．By integrating DEGs，autophagy-related genes ( ARGs) ，and WGCNA mod- ules，19 autophagy-related DEGs were identified． GO and KEGG enrichment analyses indicated that these genes were mainly involved in CD4 + T cell activation and regulation，innate immune responses，and autophagosome mem- brane formation．Immune infiltration analysis revealed that innate immune cells such as neutrophils and dendritic cells predominated during the early phase of infection，while γδ T cells and M2 macrophages became more promi- nent in the later stages．PPI network analysis identified 12 hub autophagy-related genes，among which three upreg- ulated key genes ( Eif2ak2，Ikbke，and Nfkbiz) were further confirmed．The area under the ROC curve for all three genes was 1. 000．RT-qPCR validation demonstrated significantly elevated expression of these three genes in lung tissues at 24 hours post-infection ( all P＜0. 05) ． Conclusion Eif2ak2，Ikbke，and Nfkbiz may be involved in the pulmonary infection caused by USA300 by promoting autophagy and hold promise as potential targets for immuno- therapy．

Objective:To analyze the iodine nutrition status of pregnant women in Weihai City before and after (2022, 2023) the iodine deficiency disorders (IDD) intervention, evaluate the effectiveness of intervention measures, and provide a scientific basis for adjusting IDD prevention strategies in the city.Methods:In May 2023, the intervention action of IDD was carried out for pregnent women in Weihai City. In April 2022 and December 2023, respectively, a cross-sectional survey method was conducted in four districts (cities) of Weihai City. Each district (city) was divided into five areas (east, west, south, north, and central) each year, and one township (street) was selected from each areas. At least 20 pregnant women were selected from each township (street) as survey subjects, and their household salt samples and random urine samples were collected to measure salt iodine and urinary iodine concentrations, and to compare the test results.Results:A total of 922 household salt samples were tested, including 530 iodized salt samples and 469 qualified iodized salt samples. The median salt iodine was 24.00 mg/kg. The coverage rate of iodized salt, the qualified rate of iodized salt, and the consumption rate of qualified iodized salt were 57.48%, 88.49%, and 50.87%, respectively. No significant difference was observed in medians salt iodine between 2022 (24.26 mg/kg) and 2023 (24.00 mg/kg, Z = - 1.58, P = 0.113). However, the coverage rate of iodized salt and the consumption rate of qualified iodized salt in 2023 werehigher than those in 2022 [73.72% (373/506) vs 37.74% (157/416), 65.22% (330/506) vs 33.41% (139/416), χ 2 = 120.90, 92.40, P < 0.001]. A total of 922 urine samples were collected from pregnant women, with a median urinary iodine of 135.90 μg/L. The median urinary iodine of pregnant women in 2022 was 113.55 μg/L. There was a statistically significant difference in medians urinary iodine among different regions ( H = 27.91, P < 0.001). The median urinary iodine of pregnant women in 2023 was 153.00 μg/L. There was no statistically significant difference in medians urinary iodine among different regions ( H = 3.33, P = 0.343). The medians urinary iodine of pregnant women in Huancui District, Wendeng District, and Rushan City in 2023 (156.60, 155.00, 140.85 μg/L) were higher than those in 2022 (93.60, 110.00, 110.70 μg/L), and the differences were statistically significant ( Z = - 5.44, - 4.92, - 5.99, P < 0.001). The median urinary iodine of pregnant women in Weihai City in 2023 showed a statistically significant difference compared to 2022 ( Z = - 7.62, P < 0.001). Conclusions:The IDD intervention measures in Weihai City have achieved good results, and the coverage of iodized salt, the consumption rate of qualified iodized salt, and the iodine nutrition level of pregnant women have improved. We should continue to implement intervention measures, improve the iodine nutrition level of key populations, and maintain a sustained elimination of IDD.

Objective:To analyze the iodine nutrition status of pregnant women in Weihai City before and after (2022, 2023) the iodine deficiency disorders (IDD) intervention, evaluate the effectiveness of intervention measures, and provide a scientific basis for adjusting IDD prevention strategies in the city.Methods:In May 2023, the intervention action of IDD was carried out for pregnent women in Weihai City. In April 2022 and December 2023, respectively, a cross-sectional survey method was conducted in four districts (cities) of Weihai City. Each district (city) was divided into five areas (east, west, south, north, and central) each year, and one township (street) was selected from each areas. At least 20 pregnant women were selected from each township (street) as survey subjects, and their household salt samples and random urine samples were collected to measure salt iodine and urinary iodine concentrations, and to compare the test results.Results:A total of 922 household salt samples were tested, including 530 iodized salt samples and 469 qualified iodized salt samples. The median salt iodine was 24.00 mg/kg. The coverage rate of iodized salt, the qualified rate of iodized salt, and the consumption rate of qualified iodized salt were 57.48%, 88.49%, and 50.87%, respectively. No significant difference was observed in medians salt iodine between 2022 (24.26 mg/kg) and 2023 (24.00 mg/kg, Z = - 1.58, P = 0.113). However, the coverage rate of iodized salt and the consumption rate of qualified iodized salt in 2023 werehigher than those in 2022 [73.72% (373/506) vs 37.74% (157/416), 65.22% (330/506) vs 33.41% (139/416), χ 2 = 120.90, 92.40, P < 0.001]. A total of 922 urine samples were collected from pregnant women, with a median urinary iodine of 135.90 μg/L. The median urinary iodine of pregnant women in 2022 was 113.55 μg/L. There was a statistically significant difference in medians urinary iodine among different regions ( H = 27.91, P < 0.001). The median urinary iodine of pregnant women in 2023 was 153.00 μg/L. There was no statistically significant difference in medians urinary iodine among different regions ( H = 3.33, P = 0.343). The medians urinary iodine of pregnant women in Huancui District, Wendeng District, and Rushan City in 2023 (156.60, 155.00, 140.85 μg/L) were higher than those in 2022 (93.60, 110.00, 110.70 μg/L), and the differences were statistically significant ( Z = - 5.44, - 4.92, - 5.99, P < 0.001). The median urinary iodine of pregnant women in Weihai City in 2023 showed a statistically significant difference compared to 2022 ( Z = - 7.62, P < 0.001). Conclusions:The IDD intervention measures in Weihai City have achieved good results, and the coverage of iodized salt, the consumption rate of qualified iodized salt, and the iodine nutrition level of pregnant women have improved. We should continue to implement intervention measures, improve the iodine nutrition level of key populations, and maintain a sustained elimination of IDD.

AIM:To investigate the mechanism by which wogonin(WOG)induces ferroptosis in collagen-in-duced arthritis rat fibroblast-like synoviocytes(rat CIA-FLS cells)through the nuclear factor E2-related factor 2(NRF2)/heme oxygenase-1(HO-1)signaling pathway.METHODS:Rat CIA-FLS cells were divided into:control group,low,medium,and high dose of(25,50 and 100 μmol/L)WOG group,ferroptosis inhibitor(LIP-1)group,LIP-1+high dose WOG group,HO-1 agonist cobalt protoporphyrin(COPP)group,and COPP+high dose WOG group.CCK-8 assay was used for cell viability.Crystal violet staining was used for for cell morphology.The levels of oxidative stress markers gluta-thione(GSH),malondialdehyde(MDA),and superoxide dismutase(SOD)were measured.DCFH-DA fluorescent probe was used to detect the intracellular reactive oxygen species(ROS)content as well as Western blot to detect the protein ex-pression levels of Kelch-like ECH-associated protein 1(KEAP-1),NRF2 and HO-1.RESULTS:Compared with the nor-mal control group,administration of WOG treatment resulted in a significant decrease in CIA-FLS cell viability(P<0.01),a significant increase in the level of oxidative stress(P<0.01),a significant increase in the content of ROS(P<0.01),a significant decrease in the level of expression of NRF2 and HO-1 proteins(P<0.01),and a significant increase in the level of KEAP-1(P<0.01)in the rat.Compared with the WOG group,the LIP-1-treated group showed a significant increase in cell viability(P<0.01),a significant decrease in the level of oxidative stress(P<0.01),and a significant de-crease in the content of ROS(P<0.01).Compared with the WOG group,the addition of COPP resulted in a significant in-crease in the protein expression levels of NRF2 and HO-1(P<0.01)and a significant decrease in KEAP-1 levels(P<0.01).CONCLUSION:WOG can induce ferroptosis in rat CIA-FLS cells by promoting oxidative stress through the NRF2/HO-1 signaling pathway.

Objective The molecular mechanism of Yi Lung Xuan Lung Subduing Turbidity Formula in treating rats with vascular dementia(VaD)model was investigated by network pharmacology and animal experiments.Methods The TCMSP database platform was used to screen the active ingredients and related targets of Yi Lung,Xuan Lung and Turbidity Subduing Formula.Since Maitong could not be found in the TCMSP database,the components of Maitong were retrieved by reviewing the literature and using the BATMAN-TCM Bioinformatics Analysis Tool.The GeneCards database was used to obtain VaD-related targets and predict the potential targets of Yi Lung Xuan Lung Turbidity Reducing Formula for the treatment of VaD.The network structure of"active ingredient-target intersection"of the formula was mapped by STRING and Cytoscape 3.7.2 software,and the PPI network model was established to find the key targets.GO enrichment and KEGG pathway enrichment analyses were performed on the targets of VaD treated with Yi Lung Xuan Lung Turbidity Reducing Formula.29 rats were randomly divided into the sham-operation group,the model group,the traditional Chinese medicine group and the western medicine group.The 2-VO method was used for modelling,and the water maze was used to test the memory behaviours of the rats;HE staining was used to observe the pathological changes in the hippocampal CA1 region of the animals in each group,fluorescence detection of VEGF content in the brain of the rats in each group,and ELISA to detect the content of IL-6 and TNF-α in the brain tissues of the rats in each group;Western blot was used to detect the levels of Nrf2,HO-1,P Nrf2,HO-1,P-Akt/Akt and NF-κB.Results A total of 380 targets were obtained from Yi Lung Xuan Lung Turbidity Reducing Formula,and 183 targets were intersected with VaD;The PPI network of intersected core target genes was constructed with 92 nodes and 2610 edges.The GO results suggested that the biological processes related to VaD treatment included responses to lipopolysaccharide,oxidative stress,cell migration,etc.KEGG enrichment analysis suggested that the key pathways included NF-κB,Akt,VEGF,NOD-like receptor signalling.Compared with the model group,the number of crossing platforms in the traditional Chinese medicine group was significantly increased,the histological structure and cellular morphology of hippocampal CA1 area were intact and there were few cellular degeneration,the expression levels of Nrf2,Akt,and HO-1 were significantly increased,and the expression level of NF-κB was significantly decreased(P<0.05),and inflammatory indexes,IL-6 and TNF-α,were significantly down-regulated(P<0.01),and the immunological monoclonal fluorescence showed that the vascular endothelial cell in the traditional Chinese medicine group was significantly increased,while the activity of antioxidant factor SOD increased and the content of oxidative damage factor MDA decreased(P<0.05).Conclusion The network pharmacological analysis suggested that Yi Lung Xuan Lung and Turbidity Reducing Formula could treat VaD by regulating multiple signalling pathways and biological processes.Yi Lung Xuan Lung and Turbidity Reducing Formula increased the protein expression levels of Nrf2,P-Akt/Akt and HO-1,VEGF,and improved the degeneration of neuronal cells of hippocampal CA1 area of rats with VaD,which may be through the activation of AKT/Nrf2/HO-1,and improved the lesions of hippocampal area of VaD,inhibited oxidative damage,and decreased the content of MDA,a factor that can damage oxidative processes.lesions,inhibiting oxidative damage and down-regulating the NF-κB pathway,reducing neuroinflammation,and thus improving cognitive function in VaD model rats.

Objective To establish an inhalation infection pneumonia model of C57BL/6J mice with highly virulent and multi-drug resistant Pseudomonas aeruginosa(PA)strain F291007,and to study the microbiological,pathological and immunological characteristics of this model.Methods The strain F291007 was isolated and identified before the bacterial suspension was administered to the mice via aerosolized intratracheal inoculation to establish the pneumonia infection model.In the course of infection,the conditions and survival of the mice were observed,and the bacterial loads,the histopathological states and the cytokine expression levels in the major organs were detected.Finally,three key cytokines were blocked to observe the survival of mice.Results The strain F291007 was isolated and identified.After lethal dose infection,all the mice died within 24 h.After sub-lethal dose infection,a large number of immune cells in the body were capable of phagocytosis and killing of invading pathogens,which was manifested as rapid clearance of bacteria in lungs and the exponential decrease of bacterial load with the passage of time.The pathological changes in lungs were most severe at 1 to 3 days but gradually recovered.After infection,interleukin-6(IL-6),IL-17A and tumor necrosis factor-α(TNF-α)in alveolar lavage fluid and serum were significantly increased at 1 to 3 days.After blocking of these three cytokines with specific antibodies,the survival rates of infected mice decreased significantly.Conclusion A mouse model of gradually-recovered pneumonia infection caused by PA inhalation has been established,suggesting that the first one to three days are critical to immune response after infection through multiple indicators.This mouse model can be used for research on the pathogenesis,immunoregulation and treatment evaluation of highly virulent and multi-drug resistant PA inhalation pneumonia infection.