ObjectiveTo investigate the mechanism by which the Zishen Huoxue prescription （ZSHXP） ameliorates cognitive dysfunction in rats with vascular dementia （VD） induced by the bilateral common carotid artery ligation （2-VO model rats） through regulating the glucose-regulated protein 78 （GRP78）/protein kinase R-like endoplasmic reticulum kinase （PERK）/activating transcription factor 4 （ATF4） signaling pathway. MethodsA VD rat model was established via the 2-VO method. A total of 72 male Sprague-Dawley （SD） rats were randomly divided into six groups： Sham group， Model group， donepezil hydrochloride group （0.45 mg·kg^-1）， and ZSHXP groups at low （8.90 g·kg^-1）， medium （17.80 g·kg^-1）， and high （35.60 g·kg^-1） doses，with 12 rats in each group. The Morris Water Maze test was utilized to assess spatial learning and memory abilities of rats， and the Novel Object Recognition test was used to evaluate cognitive performance. Hematoxylin-eosin （HE） and Nissl staining were applied to observe the histological and morphological changes in hippocampal tissues. Transmission electron microscopy （TEM） was used to observe the morphological changes of endoplasmic reticulum in rat hippocampal neurons. Immunofluorescence staining was adopted to detect the colocalization of neuronal nuclei antigen （NeuN） with GRP78 and βⅢ Tubulin with gasdermin D （GSDMD） in hippocampal neurons. Western blot was used to detect the expression levels of endoplasmic reticulum stress （ERS）-related proteins including GRP78， PERK， ATF4， phosphorylated protein kinase R-like endoplasmic reticulum kinase （p-PERK）， C/EBP homologous protein （CHOP）， NOD-like receptor protein 3 （NLRP3）， Caspase-1 and GSDMD. ResultsCompared with the sham operation group， the model group showed a significantly prolonged escape latency （P<0.01）， a significant decrease in the number of platform crossings and the residence time in the target quadrant （P<0.01）， and a markedly reduced recognition index （P<0.01）. Histological observations revealed that the hippocampal neurons in the model group were disorderly arranged with reduced quantity， deformed and shrunken cell bodies， and pyknotic and hyperchromatic nuclei. The number of Nissl bodies decreased significantly. The number of endoplasmic reticula reduced obviously， accompanied by abnormal dilation and swelling， and the loss of normal folding structure. The fluorescence colocalization of NeuN with GRP78 and βⅢ Tubulin with GSDMD in the hippocampus was significantly increased in the model group. The protein expression levels of GRP78， p-PERK/PERK， ATF4， CHOP， NLRP3， GSDMD and Caspase-1 in the model group were significantly elevated （P<0.01）. Compared with the model group， the donepezil hydrochloride group and the ZSHXP medium- and high-dose groups had a significantly shortened escape latency （P<0.01） and an increased number of platform crossings （P<0.05， P<0.01）. The residence time in the target quadrant was increased in the donepezil hydrochloride group and all ZSHXP groups （P<0.05， P<0.01）， with a significantly improved recognition index （P<0.01）. In the donepezil hydrochloride group and all ZSHXP groups， the number of hippocampal neurons increased with a more compact arrangement and reduced nuclear hyperchromasia. The number of Nissl bodies increased with morphological structures tending to be normal. In the ZSHXP high-dose group， the number of endoplasmic reticula increased and the folding structure was restored. The fluorescence colocalization of NeuN with GRP78 and βⅢ Tubulin with GSDMD in the hippocampus was significantly weakened in the treatment groups. In the donepezil hydrochloride group， the protein expressions of GRP78， ATF4 and CHOP were increased （P<0.01）， while the expression of p-PERK/PERK was decreased （P<0.05）. In the ZSHXP low-dose group， the expressions of GRP78， p-PERK/PERK and CHOP were elevated （P<0.05， P<0.01）. The ZSHXP medium- and high-dose groups showed a significant decrease in the protein expressions of p-PERK/PERK， ATF4 and CHOP （P<0.01）， and the high-dose group had a markedly reduced GRP78 protein expression （P<0.01）. In the donepezil hydrochloride group， the Caspase-1 protein expression was increased （P<0.01） and the NLRP3 protein expression was decreased （P<0.01）. In the ZSHXP low-dose group， the GSDMD expression was elevated （P<0.01） while the NLRP3 protein expression was reduced （P<0.01）. After treatment with medium and high doses of ZSHXP， the protein expression levels of NLRP3， GSDMD and Caspase-1 were significantly decreased （P<0.01）. ConclusionThe ameliorative effect of ZSHXP on cognitive function in 2-VO model rats may be associated with its regulation of the GRP78/PERK/ATF4 signaling pathway， which ameliorates ERS and inhibits neuronal pyroptosis.

ObjectiveTo investigate the mechanism by which the Zishen Huoxue prescription （ZSHXP） ameliorates cognitive dysfunction in rats with vascular dementia （VD） induced by the bilateral common carotid artery ligation （2-VO model rats） through regulating the glucose-regulated protein 78 （GRP78）/protein kinase R-like endoplasmic reticulum kinase （PERK）/activating transcription factor 4 （ATF4） signaling pathway. MethodsA VD rat model was established via the 2-VO method. A total of 72 male Sprague-Dawley （SD） rats were randomly divided into six groups： Sham group， Model group， donepezil hydrochloride group （0.45 mg·kg^-1）， and ZSHXP groups at low （8.90 g·kg^-1）， medium （17.80 g·kg^-1）， and high （35.60 g·kg^-1） doses，with 12 rats in each group. The Morris Water Maze test was utilized to assess spatial learning and memory abilities of rats， and the Novel Object Recognition test was used to evaluate cognitive performance. Hematoxylin-eosin （HE） and Nissl staining were applied to observe the histological and morphological changes in hippocampal tissues. Transmission electron microscopy （TEM） was used to observe the morphological changes of endoplasmic reticulum in rat hippocampal neurons. Immunofluorescence staining was adopted to detect the colocalization of neuronal nuclei antigen （NeuN） with GRP78 and βⅢ Tubulin with gasdermin D （GSDMD） in hippocampal neurons. Western blot was used to detect the expression levels of endoplasmic reticulum stress （ERS）-related proteins including GRP78， PERK， ATF4， phosphorylated protein kinase R-like endoplasmic reticulum kinase （p-PERK）， C/EBP homologous protein （CHOP）， NOD-like receptor protein 3 （NLRP3）， Caspase-1 and GSDMD. ResultsCompared with the sham operation group， the model group showed a significantly prolonged escape latency （P<0.01）， a significant decrease in the number of platform crossings and the residence time in the target quadrant （P<0.01）， and a markedly reduced recognition index （P<0.01）. Histological observations revealed that the hippocampal neurons in the model group were disorderly arranged with reduced quantity， deformed and shrunken cell bodies， and pyknotic and hyperchromatic nuclei. The number of Nissl bodies decreased significantly. The number of endoplasmic reticula reduced obviously， accompanied by abnormal dilation and swelling， and the loss of normal folding structure. The fluorescence colocalization of NeuN with GRP78 and βⅢ Tubulin with GSDMD in the hippocampus was significantly increased in the model group. The protein expression levels of GRP78， p-PERK/PERK， ATF4， CHOP， NLRP3， GSDMD and Caspase-1 in the model group were significantly elevated （P<0.01）. Compared with the model group， the donepezil hydrochloride group and the ZSHXP medium- and high-dose groups had a significantly shortened escape latency （P<0.01） and an increased number of platform crossings （P<0.05， P<0.01）. The residence time in the target quadrant was increased in the donepezil hydrochloride group and all ZSHXP groups （P<0.05， P<0.01）， with a significantly improved recognition index （P<0.01）. In the donepezil hydrochloride group and all ZSHXP groups， the number of hippocampal neurons increased with a more compact arrangement and reduced nuclear hyperchromasia. The number of Nissl bodies increased with morphological structures tending to be normal. In the ZSHXP high-dose group， the number of endoplasmic reticula increased and the folding structure was restored. The fluorescence colocalization of NeuN with GRP78 and βⅢ Tubulin with GSDMD in the hippocampus was significantly weakened in the treatment groups. In the donepezil hydrochloride group， the protein expressions of GRP78， ATF4 and CHOP were increased （P<0.01）， while the expression of p-PERK/PERK was decreased （P<0.05）. In the ZSHXP low-dose group， the expressions of GRP78， p-PERK/PERK and CHOP were elevated （P<0.05， P<0.01）. The ZSHXP medium- and high-dose groups showed a significant decrease in the protein expressions of p-PERK/PERK， ATF4 and CHOP （P<0.01）， and the high-dose group had a markedly reduced GRP78 protein expression （P<0.01）. In the donepezil hydrochloride group， the Caspase-1 protein expression was increased （P<0.01） and the NLRP3 protein expression was decreased （P<0.01）. In the ZSHXP low-dose group， the GSDMD expression was elevated （P<0.01） while the NLRP3 protein expression was reduced （P<0.01）. After treatment with medium and high doses of ZSHXP， the protein expression levels of NLRP3， GSDMD and Caspase-1 were significantly decreased （P<0.01）. ConclusionThe ameliorative effect of ZSHXP on cognitive function in 2-VO model rats may be associated with its regulation of the GRP78/PERK/ATF4 signaling pathway， which ameliorates ERS and inhibits neuronal pyroptosis.

OBJECTIVE To investigate the apoptosis-inducing effect of esculetin （Esc） on acute myeloid leukemia （AML） HL-60 cells by regulating the protein kinase B （AKT）/S-phase kinase-associated protein 2 （SKP2）/MutT homolog 1 （MTH1） pathway. METHODS AML HL-60 cells were randomly divided into control group （routine culture）， Esc low-concentration group （L-Esc group， 25 μmol/L Esc）， Esc medium-concentration group （M-Esc group， 50 μmol/L Esc）， Esc high-concentration group （H-Esc group， 100 μmol/L Esc）， and high-concentration of Esc+ SC79 （AKT agonist） group （100 μmol/L Esc+5 μmol/L SC79）. Cell proliferation in each group was detected by MTT assay and colony formation assay. The level of reactive oxygen species （ROS） in cells was measured by using the CM-H2DCFDA fluorescent probe. Cell apoptosis was analyzed by flow cytometry. Western blot assay was performed to detect the expression levels of apoptosis-related proteins [B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved caspase-3]， AKT/SKP2/MTH1 pathway-related proteins （p-AKT， AKT， SKP2， MTH1）， along with the upstream and downstream proteins of AKT phosphatidylinositol 3-kinase （PI3K）， cyclin-dependent kinase inhibitor 1 （P21） and cyclin-dependent kinase inhibitor 1B （P27）. RESULTS Compared with control group， the cell viability， colony number， and the phosphorylation levels of AKT and PI3K proteins as well as protein expressions of SKP2， MTH1 and Bcl-2 were significantly decreased （P＜0.05）， while ROS level， apoptosis rate， and the expression levels of Bax， cleaved caspase-3， P21 and P27 proteins were significantly increased （P＜0.05）. Moreover， the effects of Esc exhibited concentration-dependence （P＜0.05）. Compared with H-Esc group， above indexes of high-concentration of Esc+ SC79 group were reversed significantly （P＜0.05）. CONCLUSIONS Esc may promote massive ROS production and induce activation of apoptosis in HL-60 cells by inhibiting the AKT/SKP2/MTH1 pathway， thus inhibiting the proliferation of HL-60 cells.

Objective Rapid eye movement sleep behavior disorder（RBD） is a common sleep disorder in the elderly， and this study aims to investigate the activity of cerebral cortex based on the changes in electroencephalography （EEG） power spectrum and the difference in approximate entropy during the ictal period of RBD. Methods A total of 35 patients with idiopathic RBD who received video polysomnography monitoring were enrolled as RBD group， and 25 normal volunteers matched for age were enrolled as control group.REM EEG results with fewer artifacts was selected for both groups， and the RBD group had an increase in mandibular electromyographic activity or dream-enacting behaviors. The leads containing artifacts were excluded， and finally O1 （or O2 alternative） leads with relatively little interference were selected. After pretreatment， Fourier transform was performed for EEG data from both groups to calculate the absolute power and relative power ratio of EEG in five different frequency bands， i.e.， δ （0.5-3 Hz）， θ （4-7 Hz）， α （8-13 Hz）， β （14-30 Hz）， and γ （30-35 Hz）. The normal distribution of power values in each frequency band was tested for both groups， and the t-test was used for comparison. Approximate entropy was calculated for EEG in both two groups， and the t-test was used for comparison. Results The θ band was the dominant frequency band of REM EEG in both the control group and the RBD group. Compared with the control group， the RBD group had a significant increase in the absolute power of fast-wave activity on REM α band and significant reductions in δ/α and θ/α relative power ratios. There was a significant difference in EEG ApEn value between the control group and the RBD group （P<0.05）， and the RBD group had a higher ApEn value during REM sleep than the NC group， with a significant difference in EEG ApEn value during the phase of dream-enacting behaviors. Conclusion In the RBD group， there are significant increases in the absolute power and nonlinear approximate entropy of fast-wave activity on REM α band during the ictal period of REM， which reflects the hyperactive functional changes of cerebral cortex during the ictal period of RBD， and the involvement of cerebral cortex in RBD neural pathway disorders is an important supplement to the current theory. Moderate inhibition of cerebral cortex hyperactivity is of great significance for the treatment of RBD.

Objective To explore the outcome of improved percutaneous kyphoplasty(IPKP)for the treatment of thoracolumbar osteoporotic vertebral compression fracture(OVCF),and analyze the risk factors of postoperative bone cement leakage.Methods A total of 115 patients with thoracolumbar OVCF admitted to our hospital from March 2020 to March 2022 were included and divided into the IPKP group(n=64)and the percutaneous kyphoplasty(PKP)group(n=51)according to different surgical methods.The surgery related condition and postoperative complications of patients were recorded.The visual analogue scale(VAS)scores,Oswestry disability index(ODI)scores,anterior vertebral height,middle vertebral height and Cobb angle of patients before surgery and 3 months and 1 year after surgery were compared.The influencing factors of postoperative bone cement leakage were analyzed.Results The operation time of patients in the IPKP group was longer than that in the PKP group(P<0.05),while there was no statistically significant difference in the injected amount of bone cement during the operation between the two groups(P>0.05).The VAS and ODI scores of patients in the two groups decreased 3 months and 1 year after surgery compared with those before surgery(P<0.05),while there was no statistically significant difference in VAS or ODI scores after surgery between the two groups(P>0.05).Three months and 1 year after surgery the anterior and middle vertebral heights of patients in the IPKP and PKP groups increased and the Cobb angle decreased compared with those before surgery(P<0.05).One year after surgery,the anterior and middle vertebral heights in the IPKP group were significantly higher than those in the PKP group(P<0.05),and the Cobb angle was significantly smaller than that in the PKP group(P<0.05).Among the 115 patients with thoracolumbar OVCF,there was no complications such as infection,re-fracture,or vascular embolism occurred after surgery,and there were 13 cases with bone cement leakage,whose risk factor was incomplete endplate/posterior wall(P<0.05).Conclusion Compared with PKP,IPKP in the treatment of thoracolum-bar OVCF has better effect and better recovery of vertebral structure.Incomplete endplate/posterior wall can increase the risk of postopera-tive bone cement leakage,which requires reasonable and effective intervention measures.

Objective:To explore the categories of benefit finding among gastric cancer patients, analyze the differences and influencing factors among different groups, and provide reference for clinical nursing.Methods:A convenience sampling method was used to select 279 hospitalized gastric cancer patients admitted to the First Affiliated Hospital of Anhui Medical University from January 2024 to May 2024. The general information investigation, Benefit Finding Scale, Health-Related Hardiness Scale, Chronic Diseases Risk Perception Questionnaire and Distress Disclosure Index were used for cross-sectional survey. Latent profile analysis was used to identify the potential categories of benefit finding in patients with gastric cancer, and multivariate Logistic regression was used to analyze the related influencing factors.Results:A total of 266 valid questionnaires were returned, including 195 males and 71 females, with an age of (63.77 ± ?9.36) years. And three latent profiles of benefit finding were identified: low benefit-low growth group (31.96%, 85/266), moderate benefit group (37.59%, 100/266), and high benefit-health behavior group (30.45%, 81/266). The results of multiple Logistic regression analysis showed that compared with the moderate benefit group, the patients with course of disease<6 months ( OR = 0.344, 95% CI 0.160-0.737), cancer stage Ⅰ ( OR = 0.050, 95% CI 0.004-0.589), and highrisk perception ( OR = 0.935, 95% CI 0.878-0.996) were more likely to enter the low benefit-low growth group, and the patients without comorbidities ( OR = 2.520, 95% CI 1.250-5.081) and high self-disclosure ( OR = 1.137, 95% CI 1.007-1.283) were more likely to enter the moderate benefit group (all P<0.05). Compared with the high benefit-health behavior group, patients withcourse of disease<6 months ( OR = 0.108, 95% CI 0.039-0.301) were more likely to enter the low benefit-low growth group, male ( OR = 3.088, 95% CI 1.407-9.106), chemotherapy only ( OR = 6.515, 95% CI 2.034-20.864) and high health-related hardiness ( OR = 1.146, 95% CI 1.096-1.199) were more likely to enter the high benefit-health behavior group (all P<0.05). Conclusions:The benefit finding of gastric cancer patients has obvious classification characteristics. Clinical nursing staff should consider targeted interventions according to the characteristics of different categories of gastric cancer patients, encourage patients to face the disease with a positive attitude, and enhance patients′mental health literacy.

Objective To investigate the clinical efficacy of recombinant human granulocyte-macrophage colony-stimulating factor(rhGM-CSF)combined with bifidobacterium in the treatment of oral mucositis after chemotherapy.Methods A total of 60 post-chemotherapy patients with oral mucositis admitted to Ganzhou Cancer Hospital of Jiangxi Province from January 2023 to December 2024 were selected as subjects,the patients were divided into observation group(n=30)and control group(n=30)by using a randomized digital table method.The control group received rhGM-CSF mouthwash treatment,while the observation group was additionally administered bifidobacterium-lactobacillus triple live capsules orally.Clinical efficacy,symptom scores,inflammatory factor levels,oral microbiota indicators,and treatment safety were evaluated after 2 weeks of treatment.Results After treatment,the total effective rate in observation group was significantly higher than that in control group(P＜0.05).Post-treatment evaluations showed that patients in observation group exhibited lower scores for edema,congestion,and ulceration,along with reduced pain visual analog scores and decreased levels of inflammatory markers including Toll-like receptor 4,lectin-3,and interleukin-8,the difference were statistically significant(P＜0.05).After treatment,the oral microecological flora of patients in observation group,including lactic acid bacteria and bifidobacterium,were higher than that of control group,while porphyromonas gingivalis and fusetella were lower than that of control group,the difference were statistically significant(P＜0.05).Conclusion The clinical effect of rhGM-CSF combined with bifidobacterium in the treatment of oral mucositis after chemotherapy was significant,which could reduce various symptoms of patients,inhibit inflammatory factors,improve oral microecology,and have good safety.

The MGF300-4L protein of African swine fever virus(ASFV),a virulence-associated fac-tor,degrades IKKβ through the chaperone-mediated autophagy and enhances the stability of IKBαto suppress the generation of IL-1β and TNF-α regulated by the NF-κB signaling pathway.To iden-tify the host proteins interacting with MGF300-4L,PK-15 cells were transfected with the eukary-otic plasmid expressing MGF300-4L and analyzed using immunoprecipitation-mass spectrometry(IP-MS)to identify the host proteins that interact with MGF300-4L.Additionally,gene ontology(GO)and KEGG pathway enrichment analyses were conducted.Furthermore,molecular docking a-nalysis,co-immunoprecipitation,and laser confocal microscopy assays were performed to validate the host proteins interacting with MGF300-4L.The IP-MS analysis identified 145 host proteins that potentially interact with MGF300-4L.Subsequent GO and KEGG pathway enrichment analy-ses revealed that these proteins are predominantly involved in metabolic,cellular,and innate immune responses.Through molecular docking prediction,co-immunoprecipitation assay,and laser confocal microscopy,we identified the interaction between MGF300-4L and STAT1.This study provides critical insights into the mechanisms underlying the interactions between MGF300-4L and the host proteins.

Parkinson's disease(PD)is one of the most common neurodegenerative disorders.Recent evidence implicates pyroptosis as one of the pathogenic mechanisms in central nervous system disorders,although its specific mechanisms remain unclear.In this study,SH-SY5Y cells were transfected with py-roptosis-related proteins GSDME full-length(GSDME-F)or GSDME-N terminal(GSDME-N)plasmids revealed that GSDME-N significantly reduced mitochondrial membrane potential(P＜0.0001).To inves-tigate the mechanism by which GSDME mediates mitochondrial dysfunction,Western blotting analysis demonstrated that transfection with GSDME-N plasmids significantly increased BAX expression and en-hanced its translocation to mitochondria in both HEK 293T and SH-SY5Y cells(P＜0.05).SH-SY5Y cells treated with varying concentrations of rotenone(ROT)exhibited GSDME cleavage,elevated BAX expression(P＜0.05),increased mitochondrial BAX aggregation(P＜0.05),and reduced mitochondrial membrane potential(P＜0.01),as confirmed by Western blotting and JC-1 staining.Concurrently,MTT assays assessing cell viability and lactate dehydrogenase(LDH)release assays indicated that ROT in-duced these processes prior to pyroptosis.Furthermore,in a ROT-induced mouse PD model,ROT trig-gered GSDME cleavage,enhanced BAX expression,caused dopaminergic neuronal damage,and induced motor deficits.In summary,this study demonstrates that GSDME-N exacerbates mitochondrial damage and increases cytotoxicity by upregulating BAX expression and facilitating its mitochondrial translocation.This study provides novel insights into the role of GSDME in PD pathogenesis and suggests potential avenues for therapeutic intervention.

Objective To explore the outcome of improved percutaneous kyphoplasty(IPKP)for the treatment of thoracolumbar osteoporotic vertebral compression fracture(OVCF),and analyze the risk factors of postoperative bone cement leakage.Methods A total of 115 patients with thoracolumbar OVCF admitted to our hospital from March 2020 to March 2022 were included and divided into the IPKP group(n=64)and the percutaneous kyphoplasty(PKP)group(n=51)according to different surgical methods.The surgery related condition and postoperative complications of patients were recorded.The visual analogue scale(VAS)scores,Oswestry disability index(ODI)scores,anterior vertebral height,middle vertebral height and Cobb angle of patients before surgery and 3 months and 1 year after surgery were compared.The influencing factors of postoperative bone cement leakage were analyzed.Results The operation time of patients in the IPKP group was longer than that in the PKP group(P<0.05),while there was no statistically significant difference in the injected amount of bone cement during the operation between the two groups(P>0.05).The VAS and ODI scores of patients in the two groups decreased 3 months and 1 year after surgery compared with those before surgery(P<0.05),while there was no statistically significant difference in VAS or ODI scores after surgery between the two groups(P>0.05).Three months and 1 year after surgery the anterior and middle vertebral heights of patients in the IPKP and PKP groups increased and the Cobb angle decreased compared with those before surgery(P<0.05).One year after surgery,the anterior and middle vertebral heights in the IPKP group were significantly higher than those in the PKP group(P<0.05),and the Cobb angle was significantly smaller than that in the PKP group(P<0.05).Among the 115 patients with thoracolumbar OVCF,there was no complications such as infection,re-fracture,or vascular embolism occurred after surgery,and there were 13 cases with bone cement leakage,whose risk factor was incomplete endplate/posterior wall(P<0.05).Conclusion Compared with PKP,IPKP in the treatment of thoracolum-bar OVCF has better effect and better recovery of vertebral structure.Incomplete endplate/posterior wall can increase the risk of postopera-tive bone cement leakage,which requires reasonable and effective intervention measures.