BACKGROUND:Duhuo Jisheng Decoction is a classic prescription for the treatment of rheumatoid arthritis,but its specific mechanism is not clear.Extracellular vesicles have the powerful function of inter-cell communication and signal transmission,and may be the signal carrier for the Decoction.OBJECTIVE:To explore the effects of serum extracellular vesicles treated with Duhuo Jisheng Decoction on the proliferation,migration,invasion,and apoptosis of rheumatoid arthritis fibroblast-like synovial cells.METHODS:The rheumatoid arthritis fibroblast-like synovial cell model was established by co-culturing with tumor necrosis factor-α in vitro.The experiment was divided into five groups:normal group,model group,serum treated with Duhuo Jisheng Decoction group,extracellular vesicles treated with Duhuo Jisheng Decoction group,and extracellular vesicles treated with normal saline group.The optimal concentration and time of drug-containing serum and extracellular vesicles were screened by CCK-8 assay.Expression of inflammatory cytokines in the supernatants of cells in each group was detected by ELISA.The migration ability of rheumatoid arthritis fibroblast-like synovial cells was detected by scratch assay.The invasive ability of cells was measured by Transwell Invasion assay.Hoechst staining was adoped to detect cell apoptosis.The expression levels of apoptosis-related genes and proteins were detected by qRT-PCR and western blot assay.RESULTS AND CONCLUSION:(1)The optimal volume fraction of serum treated with Duhuo Jisheng Decoction was 10%and optimal mass concentration of extracellular vesicles treated with Duhuo Jisheng Decoction was 10 ng/mL;the optimal time for the interaction between the two was 24 hours.(2)Compared with the model group,serum treated with Duhuo Jisheng Decoction,extracellular vesicles treated with Duhuo Jisheng Decoction,and extracellular vesicles treated with normal saline could suppress the expression of inflammatory factors of rheumatoid arthritis fibroblast-like synovial cells(P<0.05),scratch healing(P<0.05),migration and invasion(P<0.05).Moreover,the inhibition of extracellular vesicles treated with Duhuo Jisheng Decoction was more significant(P<0.05).(3)Drug-containing serum and extracellular vesicles treated with Duhuo Jisheng Decoction promoted the apoptosis of rheumatoid arthritis fibroblast-like synovial cells.(4)Compared with the model group,serum treated with Duhuo Jisheng Decoction,extracellular vesicles treated with Duhuo Jisheng Decoction,and extracellular vesicles treated with normal saline could increase the expression of proapoptotic factors Caspase-3,Caspase-9,and Bax(P<0.05)and decrease the expression of antiapoptotic factor Bcl-2(P<0.05).Moreover,extracellular vesicles treated with Duhuo Jisheng Decoction had a more significant regulatory effect on apoptosis-related factors.Above findings indicate that extracellular vesicles treated with Duhuo Jisheng Decoction can inhibit the excessive proliferation,migration,and invasion of rheumatoid arthritis fibroblast-like synovial cells and promote their apoptosis.

BACKGROUND:Duhuo Jisheng Decoction is a classic prescription for the treatment of rheumatoid arthritis,but its specific mechanism is not clear.Extracellular vesicles have the powerful function of inter-cell communication and signal transmission,and may be the signal carrier for the Decoction.OBJECTIVE:To explore the effects of serum extracellular vesicles treated with Duhuo Jisheng Decoction on the proliferation,migration,invasion,and apoptosis of rheumatoid arthritis fibroblast-like synovial cells.METHODS:The rheumatoid arthritis fibroblast-like synovial cell model was established by co-culturing with tumor necrosis factor-α in vitro.The experiment was divided into five groups:normal group,model group,serum treated with Duhuo Jisheng Decoction group,extracellular vesicles treated with Duhuo Jisheng Decoction group,and extracellular vesicles treated with normal saline group.The optimal concentration and time of drug-containing serum and extracellular vesicles were screened by CCK-8 assay.Expression of inflammatory cytokines in the supernatants of cells in each group was detected by ELISA.The migration ability of rheumatoid arthritis fibroblast-like synovial cells was detected by scratch assay.The invasive ability of cells was measured by Transwell Invasion assay.Hoechst staining was adoped to detect cell apoptosis.The expression levels of apoptosis-related genes and proteins were detected by qRT-PCR and western blot assay.RESULTS AND CONCLUSION:(1)The optimal volume fraction of serum treated with Duhuo Jisheng Decoction was 10%and optimal mass concentration of extracellular vesicles treated with Duhuo Jisheng Decoction was 10 ng/mL;the optimal time for the interaction between the two was 24 hours.(2)Compared with the model group,serum treated with Duhuo Jisheng Decoction,extracellular vesicles treated with Duhuo Jisheng Decoction,and extracellular vesicles treated with normal saline could suppress the expression of inflammatory factors of rheumatoid arthritis fibroblast-like synovial cells(P<0.05),scratch healing(P<0.05),migration and invasion(P<0.05).Moreover,the inhibition of extracellular vesicles treated with Duhuo Jisheng Decoction was more significant(P<0.05).(3)Drug-containing serum and extracellular vesicles treated with Duhuo Jisheng Decoction promoted the apoptosis of rheumatoid arthritis fibroblast-like synovial cells.(4)Compared with the model group,serum treated with Duhuo Jisheng Decoction,extracellular vesicles treated with Duhuo Jisheng Decoction,and extracellular vesicles treated with normal saline could increase the expression of proapoptotic factors Caspase-3,Caspase-9,and Bax(P<0.05)and decrease the expression of antiapoptotic factor Bcl-2(P<0.05).Moreover,extracellular vesicles treated with Duhuo Jisheng Decoction had a more significant regulatory effect on apoptosis-related factors.Above findings indicate that extracellular vesicles treated with Duhuo Jisheng Decoction can inhibit the excessive proliferation,migration,and invasion of rheumatoid arthritis fibroblast-like synovial cells and promote their apoptosis.

Objective:To discuss the point-selection and point-grouping patterns and therapeutic application features in acupuncture-moxibustion treatment of acute gouty arthritis(AGA)based on complex networks and to provide references for treating AGA with acupuncture-moxibustion therapy. Methods:Articles related to acupuncture-moxibustion treatment of AGA were searched across the China National Knowledge Infrastructure(CNKI),Chongqing VIP Database(CQVIP),Wanfang Data Knowledge Service Platform(Wanfang),Chinese Biomedical Literature Service System(SinoMed),PubMed,Web of Science(WOS),and Excerpta Medica Database(EMBASE)from their inception till March 31,2023.An acupuncture-moxibustion prescription database was established after the articles were screened according to the inclusion and exclusion criteria.The association rule and complex network analyses of points were conducted using SPSS Modeler 18.1 and Gephi 0.9.7. Results:A total of 145 articles were collected,contributing 382 pieces of acupuncture-moxibustion prescriptions involving 104 points with a total frequency of 1 288.Ashi points contributed the highest frequency.The Spleen Meridian of Foot-Taiyin and the Stomach Meridian of Foot-Yangming were more commonly selected.Filiform-needle acupuncture and bloodletting therapy were more frequently used.The association rule analysis revealed that the highest degree of support belonged to"Ashi point-Zusanli(ST36)"and"Sanyinjiao(SP6)-Zusanli(ST36)",which reflected the rules of point combination of distal and proximal areas and point combination of the coupled meridians.The complex network analysis of the major points discovered a core point prescription mainly consisting of Ahi point,Zusanli(ST36),Sanyinjiao(SP6),Yinlingquan(SP9),and Taichong(LR3).Pattern differentiation and region differentiation were used in selecting adjunct points,stressing the improvements of patterns and joint-related symptoms. Conclusion:Acupuncture-moxibustion treatment of AGA follows the principle of combining major points with adjunct points selected based on pattern or region differentiation;the selection of major points focuses on regulating the deficient Zang-Fu organs,and the selection of adjunct points emphasizes improving patterns and symptoms.The specificity of therapeutic effects is also stressed.

Objective This research is to explore the Junctophilin-2 ( JPH2 ) expression in persistent atrial fibrillation with mitral valve disease.Methods The left atrial tissue samples were taken from 34 patients with mitral valve disease who underwent cardiac surgery.16 patients were in sinus rhythm, 18 patients had persistent atrial fibrillation.Western blot tech-nique was used to test the JPH2 expression, and the RT-PCR method was used to test the JPH2 gene expression.Results The Westernblot results showed JPH2 expression down-regulated in persistent atrial fibrillation group (0.94 ±0.29 vs.1.53 ± 0.61,P<0.01).However, there was no difference with JPH2 mRNA expression between atrial fibrillation patients and sinus rhythm patients(1.76 ±1.38 vs.1.15 ±0.94, P>0.05).Expression of miRNA-24 was significantly up-regulated in patients with persistent atrial fibrillation(4.49 ±4.30 vs.1.72 ±1.08, P<0.05).There was no significant difference in mean age, gender, ejection fraction and NYHA among the two groups.But the left atrial diameter was significant larger in patients with persistent atrial fibrillation compared to those in sinus rhythm(P=0.02).Conclusion Junctophilin-2 protein down-regulation may be associated with atrial fibrillation , cause left atrial remodeling and contraction dysfunction .