Background and Aims:Carotid plaque instability is a critical pathological basis for ischemic stroke.Identifying key molecular markers to evaluate plaque stability has important clinical implications.Recent studies have emphasized the regulatory roles and predictive value of long non-coding RNAs(lncRNAs)in plaque stability.In our previous transcriptome sequencing analysis of human stable and unstable carotid plaques,we identified lncRNA C-C motif chemokine ligand 3 antisense RNA 1(CCL3-AS1)as significantly upregulated in unstable plaques,suggesting a potential association with plaque instability.Therefore,this study aimed to validate CCL3-AS1 expression in an expanded plaque sample cohort and to explore its role and underlying molecular mechanism in carotid plaque destabilization.Methods:Carotid plaque specimens were obtained from patients undergoing carotid endarterectomy and classified into stable and unstable groups(n=15 per group)based on HE and Sirius red staining.qRT-PCR was used to validate the expression of candidate lncRNA CCL3-AS1.The localization and co-expression of CCL3-AS1 with macrophages in plaques were determined by RNA fluorescence in situ hybridization(FISH)combined with immunofluorescence staining.In vitro,THP1-derived macrophages were transduced with lentivirus or treated with antisense oligonucleotides(ASO)to overexpress or knock down CCL3-AS1,respectively,and the expression levels of inflammatory cytokines and matrix metalloproteinases(MMPs)were assessed.In vivo,an unstable carotid plaque model was established by tandem ligation of the right carotid artery in apolipoprotein E-deficient(ApoE-/-)mice,followed by local overexpression of CCL3-AS1.The effects on plaque morphology,macrophage infiltration,and MMP-9 expression were evaluated.Additionally,bioinformatic prediction using the catRAPID v2.1 omics platform was performed to identify potential RNA-binding proteins interacting with CCL3-AS1.RNA stability assays and RNA-binding protein immunoprecipitation(RIP)were conducted to verify the regulatory mechanism of MMP-9 expression.Results:CCL3-AS1 was significantly upregulated in unstable carotid plaques and was predominantly localized to the cytoplasm of plaque-infiltrating macrophages.In vitro,overexpression of CCL3-AS1 markedly increased the expression of MCP-1,TNF-α,IL-1β,iNOS,and MMP-9 in macrophages,whereas knockdown had the opposite effect.In the ApoE-/-mouse model of unstable carotid plaques,CCL3-AS1 overexpression led to fibrous cap rupture,increased infiltration of pro-inflammatory macrophages,enhanced MMP-9 secretion,and promoted plaque instability.Co-expression analysis revealed a strong correlation between CCL3-AS1 and MMP-9 expression(r=0.89,P=0.001).RNA stability assays demonstrated that CCL3-AS1 delayed the degradation of MMP-9 mRNA.Bioinformatic prediction identified heterogeneous nuclear ribonucleoprotein K(hnRNP-K)as a potential binding partner of CCL3-AS1.RIP and FISH co-localization confirmed the interaction,suggesting that CCL3-AS1 enhances MMP-9 mRNA stability through binding to hnRNP-K,thereby promoting its expression.Conclusion:As a macrophage-enriched inflammatory lncRNA,CCL3-AS1 may promote carotid plaque instability by enhancing MMP-9 expression via hnRNP-K-mediated mRNA stabilization.This lncRNA represents a potential molecular target for early intervention and stratification of ischemic stroke.

Background and Aims:Carotid plaque instability is a critical pathological basis for ischemic stroke.Identifying key molecular markers to evaluate plaque stability has important clinical implications.Recent studies have emphasized the regulatory roles and predictive value of long non-coding RNAs(lncRNAs)in plaque stability.In our previous transcriptome sequencing analysis of human stable and unstable carotid plaques,we identified lncRNA C-C motif chemokine ligand 3 antisense RNA 1(CCL3-AS1)as significantly upregulated in unstable plaques,suggesting a potential association with plaque instability.Therefore,this study aimed to validate CCL3-AS1 expression in an expanded plaque sample cohort and to explore its role and underlying molecular mechanism in carotid plaque destabilization.Methods:Carotid plaque specimens were obtained from patients undergoing carotid endarterectomy and classified into stable and unstable groups(n=15 per group)based on HE and Sirius red staining.qRT-PCR was used to validate the expression of candidate lncRNA CCL3-AS1.The localization and co-expression of CCL3-AS1 with macrophages in plaques were determined by RNA fluorescence in situ hybridization(FISH)combined with immunofluorescence staining.In vitro,THP1-derived macrophages were transduced with lentivirus or treated with antisense oligonucleotides(ASO)to overexpress or knock down CCL3-AS1,respectively,and the expression levels of inflammatory cytokines and matrix metalloproteinases(MMPs)were assessed.In vivo,an unstable carotid plaque model was established by tandem ligation of the right carotid artery in apolipoprotein E-deficient(ApoE-/-)mice,followed by local overexpression of CCL3-AS1.The effects on plaque morphology,macrophage infiltration,and MMP-9 expression were evaluated.Additionally,bioinformatic prediction using the catRAPID v2.1 omics platform was performed to identify potential RNA-binding proteins interacting with CCL3-AS1.RNA stability assays and RNA-binding protein immunoprecipitation(RIP)were conducted to verify the regulatory mechanism of MMP-9 expression.Results:CCL3-AS1 was significantly upregulated in unstable carotid plaques and was predominantly localized to the cytoplasm of plaque-infiltrating macrophages.In vitro,overexpression of CCL3-AS1 markedly increased the expression of MCP-1,TNF-α,IL-1β,iNOS,and MMP-9 in macrophages,whereas knockdown had the opposite effect.In the ApoE-/-mouse model of unstable carotid plaques,CCL3-AS1 overexpression led to fibrous cap rupture,increased infiltration of pro-inflammatory macrophages,enhanced MMP-9 secretion,and promoted plaque instability.Co-expression analysis revealed a strong correlation between CCL3-AS1 and MMP-9 expression(r=0.89,P=0.001).RNA stability assays demonstrated that CCL3-AS1 delayed the degradation of MMP-9 mRNA.Bioinformatic prediction identified heterogeneous nuclear ribonucleoprotein K(hnRNP-K)as a potential binding partner of CCL3-AS1.RIP and FISH co-localization confirmed the interaction,suggesting that CCL3-AS1 enhances MMP-9 mRNA stability through binding to hnRNP-K,thereby promoting its expression.Conclusion:As a macrophage-enriched inflammatory lncRNA,CCL3-AS1 may promote carotid plaque instability by enhancing MMP-9 expression via hnRNP-K-mediated mRNA stabilization.This lncRNA represents a potential molecular target for early intervention and stratification of ischemic stroke.

Iron is indispensable for the viablility of nearly all living organisms, and it is imperative for cells, tissues, and organisms to acquire this essential metal sufficiently and maintain its metabolic stability for survival. Disruption of iron homeostasis can lead to the development of various diseases. There is a robust connection between iron metabolism and infection, immunity, inflammation, and aging, suggesting that disorders in iron metabolism may contribute to the pathogenesis of arthritis. Numerous studies have focused on the significant role of iron metabolism in the development of arthritis and its potential for targeted drug therapy. Targeting iron metabolism offers a promising approach for individualized treatment of arthritis. Therefore, this review aimed to investigate the mechanisms by which the body maintains iron metabolism and the impacts of iron and iron metabolism disorders on arthritis. Furthermore, this review aimed to identify potential therapeutic targets and active substances related to iron metabolism, which could provide promising research directions in this field.

Objective To observe the effect of conductive education on gross motor function for children with cerebral palsy. Methods 98 children with cerebral palsy were divided into control group (n=48) and observation group (n=50) according to the wish of their parents.All the children received conventional rehabilitation, while the observation group accepted conductive education in addition. They were assessed with the total score of Gross Motor Function Measure (GMFM-88) before and 6 months after treatment. Results The total score of GMFM-88 was not significantly different between the groups before treatment (P>0.05), but it was different after treatment (P<0.01). Conclusion Conductive education may promote the recovery of gross motor function for children with cerebral palsy.

BACKGROUND: It has been confirmed that acidic peptide has good therapeutic effect on rat models of Alzheimer disease, but the mechanism still needs further exploration.OBJECTIVE: To observe whether acidic peptide can inhibit the production of N-methyl-D-aspartate receptor (NMDAR) and beta-amyloid (β-amyloid) in brain, and accelerate the production and excretion of nerve growth factor (NGF) in rats with Alzheimer disease.DESIGN: A randomized controlled animal experiment.SETTING: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Zhengzhou University.MATERIALS: The experiments were finished in the first laboratory of Institute of Bioactive Peptide, Zhengzhou University and the Cellular Culture Center, School of Basic Medical Sciences of Zhengzhou University from March 2005 to May 2006. Seventy 10-week-old healthy male SD rats without dementia symptoms were randomly divided into 7 groups with 10 rats in each group: normal control group, model group, saline group, glutamic acid 0.3 g/kg group, acidic peptide 15, 30 and 60 mg/kg groups.METHODS: Except the normal control group, the rats in the other 6 groups were induced into models of Alzheimer disease by damaging bilateral nucleus basalis of Meynert with ibotenic acid, and then intragastric administration of glutamic acid (0.3 g/kg) was given in the glutamic acid 0.3 g/kg group, acidic peptide of corresponding dosages in the acidic peptide 15, 30 and 60 mg/kg groups, and isovolume saline in the saline group respectively, 2 mL for each time, once a day for 20 days continuously.MAIN OUTCOME MEASURES: ① The learning ability of the rats was detected with Y-maze test immediately after the end of intragastric administration, and the times of correct responses were recorded. ② After the end of learning and memory test, the head was cut rapidly to remove brain,treated with immunohistochemical staining, and gray value was scanned with Biosens Digital Imaging System to determine the contents of NMDAR,NGF and β-amyloid in the brain of rats.RESULTS: All the 70 rats were involved in the analysis of results. ①Times of correct responses in the Y-maze test were lower in the other 6groups than in the normal control group (P ＜ 0.01), but higher in the acidic peptide 30 and 60 mg/kg groups than in the model group, saline group, glutamic acid 0,3 g/kg group and acidic peptide 15 mg/kg group (P ＜ 0.01). ②The gray values of NGF in basal forebrain in the model group, saline group,glutamic acid 0.3 g/kg group and acidic peptide 15 mg/kg group were lower than that in the normal control group (69.60±2.41, 69.62±1.46, 69.62±1.46,69.73±1.87, 80.77±2.72, P ＜ 0.01); There were no significant differences between the acidic peptide 30 and 60 mg/kg groups (79.39±2.23, 80.20±1.7, P ＞ 0.05), which were higher than the other groups. ③ The gray values of NMDAR and β-amyloid in cerebral cortex in the model group,saline group, glutamic acid 0.3 g/kg group and acidic peptide 15 mg/kg group were lower than those in the normal control group (NMDAR: 81.01±1.38, 81.31±2.06, 81.37±1.39, 79.38±1.23, 69.50±1.04; β-amyloid:74.26±1.39, 74.89±8.66, 74.88±1.46, 74.16±2.48, 67.40±3.06, P ＜ 0.01),and There were no significant differences between the acidic peptide 30and 60 mg/kg groups (P ＞ 0.05), which were lower than the other groups.CONCLUSION: Acidic peptide of 30 and 60 mg/kg can obviously ameliorate the learning and memory abilities in rat models of Alzheimer disease, which may be realized mainly through up-regulating the NGF content in basal forebrain and down-regulating the NMDAR and β-amyloid contents in cerebral cortex.