Objective : To investigate the mechanism of puerarin in the treatment of rheumatoid arthritis(RA) by network pharmacology and animal experiments. Methods : Traditional Chinese Medicine Systems Pharmcolog Database(TCMSP) and SwissTargetPrediction database were used to collect puerarin targets, and the targets of RA were obtained from GeneCards database and OMIM database. The PPI network was established by Cytoscape 3.7.2 software. Gene ontology(GO) function and Kyotoencyclopedia of genes(KEGG) enrichment analysis were performed through the Metascape database. RA rat-collagen-induced arthritis(CIA) model was reproduced using type Ⅱ collagen emulsion, 49 Wistar rats were randomly assigned to seven groups: control group, CIA model group, low-dose, medium-dose and high-dose puerarin group, methotrexate group, Tripterysium Glycosides Tablets group. Except for the control group, the other groups were given continuous gavage for 28 days after the CIA in rats model were prepared. The redness and swelling of the joints and ankle joint pathological changes were observed in each group. Western blot was used to detect the expression of Glycogen synthase kinase3β(GSK-3β), beta-catenin(β-catenin) proteins in the synovium. Real-time quantitative polymerase chain reaction(qPCR) was used to detect the expression of GSK-3β, β-catenin and c-Myc mRNA in the synovium. Results : Puerarin had 134 targets genes, RA had 5 821 target genes, and there were 102 overlapping target genes of puerarin and RA. It involved 184 signaling pathways, including JAK-STAT signaling pathway, NF-κB signaling pathway, Wnt signaling pathway, et al. The results of animal experiments showed that after the intervention of M-puerarin and MTX, the symptoms of redness and swelling of the hind foot were alleviated, the inflammatory cell infiltration in the synovium of the joint was significantly reduced, and the damage of cartilage and bone tissue was reduced. Compared with CIA group, the expressions of GSK-3β, β-catenin protein and GSK-3β, β-catenin and c-Myc mRNA in synovial tissue of rats after M-puerarin intervention decreased(P<0.05). Conclusion Puerarin has the characteristic of multi-components, multi-targets and multi-pathway intervention in RA. Puerarin may alleviate synovial hyperplasia, reduce articular cartilage erosion and bone destruction in CIA in rats by inhibiting Wnt/β-catenin signaling pathway.

Objective Given that the PI3K/AKT/mTOR signaling pathway is associated with the progression of knee osteoarthritis(KOA),this study aims to investigate whether the polarization induction of synovial macrophages mediated by the PI3K/AKT/mTOR signaling axis is the cause of KOA progression.Methods The synovial fluid of KOA KL-Ⅱ and KL-Ⅲ patients and normal individuals was collected,and the percentage of M1 macrophages(CD80,CD86)and M2 macrophages(CD163,CD206)in the synovial fluid(M1/M2 ratio)was measured to e-valuate the polarization of macrophage cytokines such as IL-1,IL-6,IL-10,and tumor necrosis factor(TNF)-α,transforming growth factor(TGF)-β Expression in KOA synovial fluid,and detect and analyze of key molecules PI3K/AKT/mTOR signaling axis PI3K,AKT3,mTORC1,and inducible nitric oxide synthase(iONS)in KOA synovial fluid.Results Compared with the synovial fluid of normal individuals,the percentage of M1 macrophages(CD80,CD86)in KOA patients increased(P<0.01),and the M1/M2 ratio increased(P<0.001);The ex-pression of IL-1,IL-6,and TNF-α in the synovial fluid of the KOA group was also higher than that of the control group(P<0.01),while the expression of IL-10 and TGF-β in the KOA group was significantly reduced(P<0.01);The key proteins PI3K,AKT3,mTORC1,and downstream inflammatory factor iONS in the PI3K/AKT/mTOR signaling pathway in the synovial fluid of the KOA group were higher than those in the control group(P<0.01).Conclusion In KOA synovial fluid,M1 macrophage polarization plays a dominant role,and the inflam-matory response mediated by M1 macrophage polarization may be the cause of synovitis.At the same time,the PI3K/AKT/mTOR signaling pathway may mediate the polarization of M1 macrophages involved in KOA inflammato-ry response.

Objective To investigate the effect of leflunomide(LEF)on the expression of associated autophagy genes in synoviocytes of rheumatoid arthritis(RA)by regulating HIF-1α signal pathway.Methods Three genera-tions of RA synovial cells were divided into blank control group,LEF group and Tripterygium wilfordii polyglyco-sides group.The blank control group was added with the same volume of DMEM culture medium.The drug group was treated with LEF(concentration 0.2 mg/ml)and Tripterygium wilfordii polyglycosides(concentration 0.03 mg/ml),the proliferation and apoptosis of synovial cells were detected by flow cytometry,the expression of IL-1 β,TNF-α,ANGPTL-4 and VEGF was detected by ELISA,the expression of HIF-1α mRNA was detected by qRT-PCR,and the expression of HIF-1 α,Beclin-1 and BNIP3 protein was detected by Western blot.Results Com-pared with Tripterygium wilfordii polyglycosides group,the expression of IL-1 α,TNF-α,ANGPTL-4 and VEGF in synovial supernatant of LEF group decreased;compared with the blank control group,the expression of HIF-1αmRNA in synovial cells of LEF group and Tripterygium wilfordii polyglycosides group decreased,and the effect of LEF group was the most obvious;compared with the blank control group,the protein expressions of HIF-1α,Bec-lin-1 and BNIP3 in synovial cells of LEF group and Tripterygium wilfordii polyglycosides group decreased,and the effect of LEF group was the most obvious.Conclusion LEF can inhibit the expression of inflammatory factors in RA synovial cells,inhibit HIF-1α signaling pathway,inhibit the expression of autophagy-related genes Beclin-1 and BNIP3,and improve the pathological state of synovitis.

Objective: To observe the effects of the tofacitinib on interstitial lung disease( ILD) by regulating the JAK⁃STAT signaling pathway. Methods: Wistar rats were randomly divided into 4 groups:normal group , ILD group ,prednisone acetate group , and tofacitinib group. Except for the normal group , the other three groups were given 3mg/ml bleomycin solution for modeling. After 28 days of intragastric administration , the lung tissues of all rats were collected for hematoxylin⁃eosin staining ( HE) and Western blot ( WB) to detect the protein levels of JAK1 and STAT1;Enzyme⁃linked immunosorbent assay(ELISA) was used to detect tumor necrosis factor in rat serum (TNF) Ⅳ α , interleukin (IL) Ⅳ6 , IL⁃10 , IL⁃1β . Results : HE staining of lung tissue in ILD ,prednisone acetate group and tofacitinib group showed alveolar tissue thickening , alveolar wall capillary congestion , bronchial luminal epithelial cells shedding, and inflammatory cell exudation. The results of WB showed that JAK1 and STAT1 significantly increased in ILD group , and decreased in different degrees compared with ILD group , tofacitinib group and prednisone acetate group (P < 0. 05) . The ELISA results showed that the expressions of serum TNF⁃α , IL⁃6 and IL⁃1β in the ILD group were significantly higher than those in the normal group (P < 0. 01) . The expression of pine group decreased (P < 0. 05) , and the expression of IL⁃10 was the opposite. Conclusion Tofacitinib reduces lung tissue damage and the inflammatory response in the treatment of ILD by inhibiting the JAK⁃STAT pathway and down⁃regulating the expression of inflammatory factors TNF⁃α , IL⁃6 , IL⁃1β and up⁃regulating the anti⁃inflammatory factor IL⁃10.

ObjectivesTo retrospectively analyze the clinicopathological features of neuroblastoma (NB) and investigate the signiifcance of abnormality ofN-myc and anaplastic lymphoma kinase (ALK) gene copy number change as well asALKmu-tations in NB.Methods Eighty-three NB patients were collected and classiifed into different subgroups according to the clinical stage and histology. Fluorescence in situ hybridization (FISH) was performed to detect the abnormalities ofN-mycandALK genes. The extracted DNA was ampliifed by PCR and sequenced to investigate the point mutations of theALK gene. Follow-up data were collected and survival analysis was performed.ResultsFISH detection showed that the aberration ofN-mycgene copy number presented as gain and ampliifcation. The aberration ofALK gene presented as point mutation and gain. It was shown that 17 cases had the abnormality of bothN-myc andALK gene. Survival analysis showed that the prognostic factors included the clinical stage, age and abnormality ofN-myc genes.ConclusionDetection ofN-myc andALK abnormality in NB would be helpful for evaluating the prognosis and providing theoretical basis forALK target therapy.

Objective To study the role of peripheral blood T regulatory cells (Treg) markers, as well as different Treg cells in the pathogenesis of systemic lupus erythematosus (SLE) and explore the correla-tion betweenCD127 and Foxp3. The immunosuppressive effect of CD4~+CD25~+CD127~(low/-) T cell is explored. Methods ①Four-color direct fluorescence-labeled and multi-parameter flow cytometry were used to detect peripheral blood CD4~+CD25~+ T cells and other Treg cells accounted for the proportion of CD4~+ T cells in 40 SLE patients (19 cases with active disease 21 cases in remission) and 15 healthy controls. Meanwhile, its correlations with anti-dsDNA antibodies among 7 groups were analyzed.②Flow cytometry sorting combined with cell culture were applied to detect and analyze the proliferation inhibition effect of CD4~+CD25~+CD127~(low/-)regulatory T cells to CD4~+CD25~-effector T cell.Two independent samples t test,ANOVA for repeated measures,Pearson's correlation and Spearman's correlation were used for statistical analysis.Results ①The cell ratio of the 7 groups of SLE patients was(6.1±1.7)%,(3.1±1.3)%,(2.1±1.0)%,(1.6±0.3)%(0.97±O.28)%,(0.69±0.23)%and(O.71±0.35)%respectively.In the SLE group,the proportion of the first 6 groups was lower than the control group (P<0.05).For CD4~+CD127~(low/-)Foxp3~+,there wag no difference in the two groups(P>O.05).②CD4~+CD25~+Foxp3~+,CD4~+CD25~(high)Foxp3~+ T cells and IgA ratio Was positivelv correlated.While CD4~+CD25~(high)CD127~(low/-) T cells and anti-SSB antibodies was positively correlated in SLE patients.③The seven group of cells,in addition to CD4~+CDl27~(low/-)Foxp3~+ T cell ratio were lower in early-onset SLE patients than those in patients at remission(P<0.05).④The cell propor-tion of the first 6 groups was lower than that of post steroid treatment(P<0.05).⑤Foxp3 expression of CD4~+CD25~+T ceils and CD4~+CD25~(high) T cells was positively correlated with low expression of CDl27 in the early onset group,remission group and the control group.⑥The CD4~+CD25~- effect T cells could be suppressed by their own CD4~+CD25~+CDl27~(low/-) regulatonry T cells in vitro,and the inhibition of SLE patients was significantly lower than controls.Conclusion The immunological abnormality of SLE may be associated with the qnatity and functional defects of immune regulatory T cells.CDl27 may be a possible alternative to Foxp3 regulatory T cell-specific surface markers.

Malignant hyperthermia is a rare autosomal-dominant genetic disorder. This article would review the advance in the researchs of malignant hyperthermia, including its incidence, pathogenesis and diagnosis.

In recent years,many attentions have been attracted by statin-associated neuromuscular complications.This paper reviewed the incidence,pathogenesis,risk factors,and management of statin-associated neuromuscular complications.

Objective To investigate the clinical features and characteristic of auxiliary examination of neuralgic amyotrophy(NA).Methods8 patients with idiopathic neuralgic amyotrophy were analyzed retrospectively with literature review.Results and ConclusionAntecedent events were found before attack in 5 patients.Serious pain,which was followed by paresis and atrophy,as initial symptom,was found in 7 patients.Damage in the upper and middle trunk distribution occurred most frequently.Elevated liver enzymes were found during the early phase of their attacks in 2 patients.CSF analysis was performed in 5 patients,and mildly elevated protein was found in two.5 patients had been misdiagnosed as cervical spondylosis.After medical and/or physical therapy(6 patients with corticosteroid and physical therapy,2 only with physical therapy),all the patients had been followed up for 2～13 months.Muscle power was recovered in 67% of all the patients,partially recovered in 33%.

Objective To observe the clinical efficacy of amitriptyline with manipulative therapy for episode tension-type headache (ETTH). Methods 62 patients were randomized into 2 groups: amitriptyline group, amitriptyline with manipulative therapy group. Numbers of days with headache(NDH) and days with medication(NDM), intensity of the headache(IH) between the two weeks before randomisation and 1～2, 3～4 weeks after randomization were recorded by participants in headache diaries. Results In the first 2 weeks after treatment, the NDH in amitriptyline group and amitriptyline with manipulative therapy group were (6.00±2.21) d and (4.19±1.70) d respectively (P<0.05), while in the second 2 weeks, they were (4.00±1.86) d and (2.16±1.21) d respectively (P<0.05). The NDM in amitriptyline group and amitriptyline with manipulative therapy group were (1.77±1.23) d and (1.32±1.22) d respectively in the first 2 weeks after treatment (P<0.05), but there was no significantly difference between the two groups in the second 2 weeks. The IH (with Visual Analog Scale) were(3.32±0.94) and (2.39±1.33)respectively (P<0.05) in the second 2 weeks. Conclusion Amitriptyline with manipulative therapy is better than amitriptyline alone in reducing the symptoms of episode tension type headache.