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.

Objective To investigate the in vitro metabolites and metabolic pathways of phenethylamine new psychoactive substances 2C-B-FLY and 25B-NBOH in human liver microsomes.Methods The human liver microsome incubation model was established,and the samples were centrifuged,blow-dried and re-dissolved after 3 h of co-incubation in a water bath at 37℃.The samples were detected by ultra-high performance liquid chromatography Q Exactive mass spectrometry in ESI+mode,and the parent drug and its metabolites were examined by applying Full scan-ddMS2 data-dependent acquisition mode.Results After in vitro incubation with human liver microsomes,2C-B-FLY was metabolised to produce a total of four phase I metabolites and one acetylated phase II metabolite,which underwent metabolic reactions such as hydroxylation,debromination,and N-acetylation.25B-NBOH was metabolised to produce a total of eight phase I metabolites and two glucuronidated phase II metabolites,which comprised the major biotransformation reactions of O-demethylation,hydroxylation,amine dehydrogenation,N-dealkylation and glucuronidation.Conclusion This study describes the metabolites and metabolic pathways of 2C-B-FLY and 25B-NBOH in a human liver microsomal incubation system,providing a scientific basis for the in vivo detection of new psychoactive substance of the phenethylamine group.

Antibodies, Antinuclear ; Autoantibodies ; China ; Cohort Studies ; Humans ; Scleroderma, Systemic

Objective To investigate the role of Power Arm in en-masse retraction of maxillary anterior teeth using clear aligner (CA) and micro-implant anchorage (MIA). Methods The three-dimensional (3D) model of maxillary anterior teeth by combined use of CA and MIA was established, and the 6 mm-height Power Arm, was attached to the canine or appliance. The initial displacement and the maximum von Mises stress of periodontal ligament under three loading conditions were analyzed, namely the force was loaded by CA+150 g retraction force at canine, CA+150 g retraction force on Power Arm at appliance, CA+150 g retraction force on Power Arm at canine. Results In sagittal direction, the crown and root displacement difference of maxillary central incisor was 129, 129, 133 μm，respectively. The crown displacement of the maxillary first molar was -23.3, -23.5, -26.8 μm, respectively. The maximum von Mises stress of periodontal ligament in central incisor was 48.4, 72.6, 40.0 kPa, respectively, and that of the first molar was 5.3, 10.5, 5.8 kPa, respectively. Conclusions It can not be testified that retraction of the 6 mm-height Power Arm at canine or appliance with 5 mm-height mini-screw has more advantages than retraction of the canine directly for more favorably controlling the torque of incisors, saving anchorage of posterior teeth and decreasing von Mises stress of the periodontal ligament.

Objective To analyze the roles of B-cell ccaffold protein with ankyrin repeats 1 (BANK1) in collagen-induced arthritis (CIA) murine model and the correlation with disease severity.Methods CIA murine model were established and evaluated.In different disease stages,the serum levels of anti-C Ⅱ auto-antibodies and BANK1 were detected by electrochemiluminescence immunoassay(ELISA).Moreover,the expression of BANK1 mRNA in peripheral blood cells were detected by real-time polymerase chain reaction (PCR) and its correlation with clinical scores was analyzed.Then the percentage of BANK1 expression in B cells in spleen and draining lymph nodes were detected by flow cytometry and the level of BANK1 protein in spleen was detected by Western blotting according to the results afore mentioned.The data was analyzed by Statistical Product and Service Solutions (SPSS) Stastistics 21.0 and figures were made with Graph Pad Prism 6.Repeated measure ANOVA was used to assess differences between the two groups.Correlations were analyzed by Spearman correlation analysis.Linear regression analysis was done when a correlation was identified.Results The incidence of CIA was over 90％.The clinical scores of arthritic mice was positively correlated with the serum levels of anti C Ⅱ total IgG antibody (r=0.717 5,P ＜0.01),anti C] IgG2a antibody (r=0.675 3,P＜0.01) and anti C Ⅱ IgG2b antibody (r=0.889 4,P＜0.01) respectively.The BANK1 level in the serum and the BANK1 mRNA expression were significantly decreased in different disease stages in CIA mice when compared with normal mice.The negative correlation between the BANK1 mRNA expression and clinical scores (r=-0.485 4,P＜0.01) was observed.The percentage of BANK1 +CD19+ cells in spleen and draining lymph nodes and the level of BANK1 protein in spleen were reduced in CIA as well.Conclusion Along with the disease progress in CIA,BNK1 expression is declined,which weakens the negative regulation of BANK1 on B cells.This change goes hand in hand with the severity of arthritis.

ATP-Binding Cassette Transporters ; genetics ; DNA Copy Number Variations ; genetics ; Genetic Predisposition to Disease ; Gout ; genetics ; Humans ; Receptors, IgG ; genetics ; Receptors, Interleukin-17 ; genetics

Objective To evaluate the efficacy and safety of aripiprazole treatment for co-morbid attention defi?ciency hyperactivity disorder (ADHD) in children with Tourette syndrome (TS). Methods Forty four TS children with co-morbid ADHD were randomly divided into aripiprazole group and haloperidol group. The aripiprazole group and halo?peridol group received aripiprazole and haloperidol treatment for 12 weeks, respectively. Yale global tic severity scale (YGTSS) and Conners parent symptom questionnaire (PSQ) were used to assess the tic and ADHD symptoms before, 2, 4, 8 and 12 weeks after treatment. Side effects were recorded weekly. Results Repeated measure ANOVA indicated that the main effects of groups was not significant to the YGTSS scores (P>0.05), but significant to the PSQ scores (P<0.05). After 12-week treatment, the YGTSS scores between two groups were not significantly different (P>0.05). The PSQ scores of aripiprazole group were significantly lower than that of haloperidol group. The adverse reactions of aripiprazole group were milder compared with the haloperidol group (P<0.05). Conclusions The present study demonstrates that aripipra?zole has the same efficacy in the treatment of tics as haloperidol, improves co-morbid ADHD symptoms, and its adverse reactions are much less compared with haloperidol.

Objective To investigate the current status of family rehabilitation for children with disability and the unmet needs of habilitation/rehabilitation for their parents. Methods Family Rehabilitation Needs Survey Questionnaire was used to investigate 186 families of the children with disability from 5 rehabilitation institutions in 4 provinces. Results Most families accepted family rehabilitation, including family support, developed rehabilitation scheme, telephone counseling, real-time guidance and appointment for family rehabilitation. Most families would spent 1-2 hours a day for the family rehabilitation, and prefer the rehabilitation of gross motion. However, the rehabilitation agencies played less importance on supports of family rehabilitation. Conclusion The family rehabilitation supports needs to be improved in specification of rehabilitation process, developing rehabilitation scheme and quality control.

Febuxostat is a new type of selective xanthine oxidase inhibitor,which mainly be used for the treatment of hyperuricemia patients with gout symptoms. The recommended initial dose of febuxostat is 40 mg once daily. In the present,there is no sufficient evidence to demonstrate that the clinical effects of febuxostat in reducing the uric acid are better than that of allopurinol. However,it is reported that febuxostat in 80 mg has better treatment effects in gout patients with diabetes or≥65 years old. The common adverse reactions of febuxostat are liver dysfunction,diarrhea,headache,nausea,rash,and so on. The differences of adverse reactions in cardiovascular system between febuxostat and allopurinol are not statistically significant.