Clinical Efficacy of Compound Tripterygium wilfordii Preparation（Xinfeng Capsules） in Improving Immune Inflammation of Rheumatoid Arthritis Due to Spleen Deficiency and Dampness Exuberance Based on Apriori Algorithm and Random Walk Model / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Xinfeng capsules on immunoinflammatory indicators in patients with rheumatoid arthritis （RA） due to spleen deficiency and dampness exuberance. MethodA total of 102 patients were randomly divided into control group and observation group according to the random number table method， with 51 cases in each group. All patients were treated with methotrexate tablets， while those in the observation group received additional Xinfeng capsules. The course of treatment in both groups was 12 weeks. The 28-joint disease activity score （DAS28）， visual analogue scale （VAS） scores， morning stiffness time， erythrocyte sedimentation rate （ESR）， high-sensitivity C-reactive protein （hs-CRP）， rheumatoid factor （RF）， anti-cyclic citrullinated peptide （anti-CCP） antibody， vascular endothelial growth factor （VEGF）， and serum amyloid A （SAA） of the two groups before and after treatment were compared. The efficacy and incidence of adverse events were compared between the two groups. The Apriori association rule model and random walk model were constructed to evaluate the effect of Xinfeng capsules in improving hs-CRP， ESR， RF， SAA， VEGF， and anti-CCP. ResultThere were no dropouts in this study. There was no statistical difference in the indicators between the two groups before treatment. After 12 weeks of treatment， the total effective rate in the observation group was 90.19% （46/51）， which was higher than 74.51% （38/51） in the control group （χ2＝4.320，P<0.05）. DAS28， VAS score， and morning stiffness time in the observation group were improved compared with those in the control group （P<0.05）. Apriori association rule model results showed that the application of Xinfeng capsules in the observation group had a strong correlation with the reduction of RF， ESR， hs-CRP， SAA， and VEGF. The results of the random walk model showed that the improvement coefficients of hs-CRP， ESR， RF， SAA， and VEGF in the observation group were all better than those of the control group， and the improvement coefficient of anti-CCP in the control group was better than that of the observation group. The improvement degree of hs-CRP， ESR， RF， SAA， and VEGF in the observation group was superior to that of the control group （P<0.05）. The incidence of adverse reactions in the observation group was lower than in the control group （χ2＝4.057，P<0.05）. ConclusionOn the basis of the treatment with methotrexate tablets， Xinfeng capsules can effectively improve the immunoinflammatory level in RA due to spleen deficiency and dampness exuberance and reduce the incidence of adverse reactions.

Mechanism of Xinfeng Capsules improving rheumatoid arthritis based on CD19~+B cells regulating FAK/CAPN/PI3K pathway / 中国中药杂志

To observe the effect of Xinfeng Capsules on rheumatoid arthritis (RA) B lymphocytes,inflammatory mediators,FAK/CAPN/PI3K pathway,in order to explore the mechanism of Xinfeng Capsules in improving clinical symptoms of RA.Joint and systemic symptoms of RA patients were observed,and laboratory indicators[hemoglobin (HGB),platelet count (PLT),erythrocyte sedimentation (ESR),immunoglobulin (Ig) G,Ig A,Ig M,rheumatoid factor (RF),anti-cyclic citrulline antibody (CCP-AB),C-reactive protein (CRP)]were detected.ELISA was used to detect serum interleukin (IL)-1β，IL-10,IL-33,chemokine 5 (CCL5),and vascular endothelial growth factor (VEGF).CD3~-CD19~+B cells were measured by flow cytometry.Western blot was used to detect FAK,p-FAK,CAPN,PI3K protein.The results showed that Xinfeng Capsules could significantly alleviate RA joint and systemic symptoms and improve clinical efficacy.And Xinfeng Capsules could increase HGB,decrease PLT,CCP-AB,CRP,ESR index,upregulate IL-10 expression,and down-regulate IL-1β，IL-33,CCL5,VEGF,CD3~-CD19~+B cells,FAK,p-FAK,CAPN,PI3K expressions (P<0.01).Based on the above results,Xinfeng Capsules may reduce the expression of CD3~-CD19~+,regulate the balance of inflammatory cytokines and chemokines,inhibit abnormal activation of FAK/CAPN/PI3K pathway,and improve clinical symptoms of RA.

Effect of Xinfeng Capsules-containing serum on TNF-α-induced apoptosis and inflammation of fibroblast-like synoviocytes in rheumatoid arthritis / 中国中药杂志

The aim of this paper was to observe the effect of Xinfeng Capsules(XFC)-containing serum on the apoptosis and inflammation of fibroblast-like synoviocytes(FLS) in rheumatoid arthritis(RA) induced by tumor necrosis factor-α(TNF-α), so as to investigate the mechanism of XFC in the treatment of RA. RA-FLS immortalized cell line was established, and XFC drug-containing serum was prepared. CCK-8, ELISA, RT-qPCR, immunofluorescence and TUNEL were used to observe the effect of XFC-containing serum on RA-FLS apoptosis and inflammatory indexes. CCK-8 results showed that the optimal concentration and time of TNF-α on RA-FLS were 10 ng·mL~(-1) and 48 h, respectively; and the optimal concentration and time of XFC on RA-FLS were 6.48 mg·g~(-1) and 72 h, respectively. The results of ELISA showed that compared with RA-FLS group, the expressions of TNF-α, IL-1β, IL-6, IL-8 in TNF-α+RA-FLS group were significantly increased, while the expressions of IL-4 and IL-10 were significantly decreased(P<0.01); after intervention with XFC-containing serum, the expressions of TNF-α, IL-1β, IL-6, IL-8 were significantly decreased, whereas the expressions of IL-4 and IL-10 were significantly increased(P<0.01). The results of RT-qPCR showed that compared with RA-FLS group, the mRNA expressions of Fas, FasL, caspase-3, caspase-8, Bax, Bcl-X1 in TNF-α+RA-FLS group were significantly decreased, while the mRNA expression of Bcl-2 was significantly increased(P<0.001); after intervention with XFC-containing serum, the mRNA expressions of Fas, FasL, caspase-3, caspase-8, Bax, Bcl-X1 were significantly increased, whereas the mRNA expression of Bcl-2 was significantly decreased(P<0.01). The results of immunofluorescence showed that compared with RA-FLS group, the protein expressions of caspase-3 and Bax in TNF-α+RA-FLS group was significantly lower than those in RA-FLS group(P<0.05); after intervention with XFC-containing serum, the protein expressions of caspase-3 and Bax were significantly increased, whereas the protein expression of Bcl-2 was significantly decreased(P<0.05). TUNEL results showed that compared with RA-FLS group, the apoptosis of TNF-α+RA-FLS group was decreased(P<0.05); after intervention with XFC-containing serum, the apoptosis was significantly increased(P<0.05). One of the mechanisms of XFC in the treatment of RA is to promote the apoptosis of RA-FLS and inhibit its inflammatory reaction.

Xinfeng Capsules promotes apoptosis of synovial fibroblasts and attenuates inflammation in rheumatoid arthritis by regulating lncRNA MAPKAPK5-AS1 / 中国中药杂志

To explore the regulatory effects of Xinfeng Capsules(XFC) on the apoptosis of synovial fibroblasts(FLS) and inflammation in rheumatoid arthritis(RA) via lncRNA MAPKAPK5-AS1(MK5-AS1). Thirty healthy people and 30 patients with RA due to spleen deficiency and dampness exuberance were collected for extracting the peripheral blood mononuclear cells(PBMCs) before and after XFC treatment, which were used to observe the correlation between MK5-AS1 and clinical indicators as well as MK5-AS1 expression before and after XFC treatment. Following the establishment of RA-FLS cell line and the preparation of XFC-containing serum, MK5-AS1-overexpression plasmid was constructed and transfected into RA-FLS for investigating the efficacy of XFC-containing serum in regulating inflammation and apoptosis of RA-FLS via MK5-AS1. The expression of MK5-AS1 in PBMCs of patients with RA due to spleen deficiency and dampness exuberance was decreased(P<0.001). The ROC curve analysis revealed the AUC of 83.9%. Correlation analysis showed that MK5-AS1 was negatively correlated with ESR, CRP, RF, CCP, and spleen deficiency and dampness exuberance syndrome score. The expression of MK5-AS1 increased significantly after XFC treatment(P<0.001). As demonstrated by association analysis, XFC decreased MK5-AS1, ESR, CRP, RF, and spleen deficiency and dampness exuberance syndrome score, with the degree of support all greater than 83%, confidence greater than 80%, and lift greater than 1. The results of RT-qPCR showed that the MK5-AS1 RNA expression significantly decreased after TNF-α stimulation(P<0.01), which, however, increased significantly after the intervention with XFC-containing serum(P<0.05). Such expression rose again after the transfection of pcDNA3.1-MK5-AS1(P<0.01). ELISA results showed that TNF-α stimulation elevated the expression of pro-inflammatory factor IL-17 but lowered the expression of anti-inflammatory factor IL-4(P<0.01). After intervention with XFC-containing serum, the expression of IL-17 decreased while that of IL-4 increased(P<0.01). The transfection of pcDNA3.1-MK5-AS1 contributed to the reduction in IL-17 expression but the elevation in IL-4 expression(P<0.01). The immunofluorescence(IF) findings demonstrated that the expression of pro-apoptotic protein Bax was down-regulated, whereas that of the anti-apoptotic protein Bcl-2 was up-regulated after TNF-α stimulation(P<0.01). After the intervention with XFC-containing serum, the Bax expression was increased, while Bcl-2 expression was decreased(P<0.01), which were remarkably collaborated by the transfection of pcDNA3.1-MK5-AS1(P<0.05). The expression of MK5-AS1 is significantly decreased in both RA-PBMCs and RA-FLS, implying that XFC inhibits inflammatory reaction and promotes the apoptosis in RA by regulating the expression of MK5-AS1.

Effect of traditional Chinese medicine invigorating spleen unit therapy on inflammatory markers in patients with osteoarthritis based on random walk model / 中国中药杂志

To evaluate the effect of traditional Chinese medicine(TCM) invigorating spleen unit therapy on inflammatory markers of osteoarthritis(OA) patients by random walk model. The patient information was collected by the data processing system of medical records of the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine. In-patient information of the department of rheumatology of the hospital between June 2012 and December 2016 was collected and summarized. Based on the use of traditional Chinese medicine decoction and hospital-prepared compound Qiyi Capsules(Xinfeng Capsules),the patients were divided into the unit therapy group and the simple endotherapy group. The random walk model was used to evaluate the effect of traditional Chinese medicine invigorating spleen unit therapy on TCM spleen therapy unit(ESR) and high sensitive C reactive protein(hs-CRP). A total of 3 517 cases of OA patients met the study requirements. The simple endotherapy group had 1 771 cases(50. 36%),while the unit therapy group had 1 746 cases(49. 64%). The baseline data analysis showed that the general information of the cases,the TCM oral intake frequency and the core prescription information had no statistically significant difference,with comparability. The unit therapy group showed the maximum ESR stochastic volatility at 924,walking step number of 1 771,forward walking growth rate at 0. 264 5,ratio at3. 78,random fluctuation power law at 0. 306 5± 0. 076 8,positive increase rate of comprehensive evaluation index at 0. 264 5,and comprehensive evaluation index record number of 1 771; whereas the simple endotherapy group showed the maximum ESR random fluctuation value at 478,walking step number of 1 399,forward walking growth rate at 0. 152 4,ratio at 6. 56,random fluctuation power law at 0. 347 4±0. 101 7,positive increase rate of comprehensive evaluation index at 0. 152 4,and comprehensive evaluation index record number of 1 399. The unit therapy group showed the maximum hs-CRP random fluctuation value at 391,walking step number of1 081,forward walking growth rate at 0. 178 1,ratio at 5. 62,random fluctuation power law at 0. 343 6±0. 094 7,positive increase rate of comprehensive evaluation index at 0. 178 1,and comprehensive evaluation index record number of 1 081; while the simple endotherapy groups showed the maximum hs-CRP random fluctuation value at 210,walking step number of 797,forward walking growth rate at0. 113 2,ratio at 8. 83,random fluctuation power law at 0. 382 6±0. 109,positive increase rate of comprehensive evaluation index at0. 113 2,and comprehensive evaluation index record number of 797. According to our department of rheumatism,there was a longrange correlation between the two groups in the comprehensive evaluation index and the intervention measures. TCM spleen strengthening unit therapy has a better effect in alleviating the inflammatory index of OA than traditional Chinese medicine.

Quantitative research on toxic component-wilforlide A in Xinfeng capsules before and after compatibility / 中国基层医药

Objective To study the content changes of wilforlide A in Xinfeng capsules before and after compatibility.Methods The content of wilforlide A was determined in Xinfeng capsules ester content and in Tripterygium wilfordii Hook.f.single solution by TLCS method.Developing solvents:chloroform and methanol (85:1),scanning wavelength:230 nm.Results Wilforlide A showed a good linear relationship in 1.700-5.100p g,the regression equation is Y =1 118.766 + 3 253.751 X (r =0.999).The average recovery of the wilforlide A in Xinfeng capsules was 102.84％,and RSD was 3.64％.The average recovery of the wilforlide A in the Tripterygium wilfordii Hook.f.single decoction was 98.14％,and RSD was 2.21％,compatibility levels before and after the change (compatibility with 0.179mg/g,0.136mg/g after compatibility).Conclusion The established TLCS method was simple,specific and accurately quantitative.The comparative study found that the compatibility of Xinfeng capsules Tripterygium reduced the content of wilforlide A,which may be related to decreasing toxicity.