[Effect of Xinfeng Capsule on Related Factors of Thrombus Formation and Inflammatory Cytokines in Active Ankylosing Spondylitis Patients].

Objective To observe the effect of Xinfeng Capsule (XFC) on related factors of thrombus formation and inflammatory cytokines in active ankylosing spondylitis (AS) patients. Methods Seventy-six active AS patients were assigned to the XFC group and the Sulfasalazine treated group (SASP group) , 38 in each group according to random digits table. Patients in the SASP group took SASP, 0. 25 g per tablet, 4 tablets each time, twice per day. Those in the XFC group took XFC, 0. 5 g per pill, 3 pills each time, three times per day. All medication lasted for 12 successive weeks. Platelet count and coagulation functions were determined. Factors of thrombus formation [including thromboxane B2(TXB2), prostaglan- din 12 (PG12), 6-ketone-prostaglandin F1 (6-keto-PGF1) , platelet granular membrane protein140 (GMP140), plasminogen activator inhibitor 2 (PAI-2 ) ], erythrocyte sedimentation rate (ESR), C reactive protein (CRP) , and levels of cytokines (TNF-α, IL-4, IL-10, IL-17) were detected. mRNA expressions of nuclear factor activator (Act1) , NF-κB inhibitory protein-alpha (IKBα) , inhibitor of kappa-B kinase beta (IKKß) , NF-κB protein 65 (NF-κB/P65), and NF-κB protein 50 (NF-κB/P50) were detected by real-time fluorescent quantitative PCR (RT-PCR). Meanwhile, the protein expression of NF-κB/P65 and NF-κB/P50 were detected by Western blot. Results Compared with before treatment in the same group, levels of PLT, fibrinogen (FBG), D-dimer (DD), TXB2, GMP140, and PAI-2 were significantly decreased, but 6-keto-PGF1 level was significantly increased in XFC group after treatment (P <0. 01). Besides, the improvement of above indices was significantly superior in the XFC group to SASP group in the same period (all P <0. 01). Compared with before treatment and SASP group after treatment, IL-17 level was significantly decreased, IL-4 and IL-10 were significantly increased, levels of ESR and CRP decreased in the XFC group after treatment (P <0. 05, P <0. 01). Compared with before treatment in the same group, mRNA expressions of Act1, IKKß, IKBα, NF- κB/P50, and NF-κB/P65, and protein expressions of NF-κB/P65 and NF-κB/P50 were obviously reduced in the two groups after treatment (P <0. 05, P <0. 01). Besides, mRNA expressions of lKKß, IKBα, NF-κB/ P50, and NF-κB/P65, and protein expressions of NF-κB/P65 and NF-κB/P50 were more obviously reduced in the XFC group than in the SASP group (P <0. 05, P <0. 01). Conclusions XFC could improve thrombosis re- lated factors in AS patients. Its mechanism might be associated with regulating cytokines and inhibiting ex- cessive activation of NF-κB signal pathway.

[Efficay Observation of Xinfeng Capsule for Treating 58 Cases of Sjogren's Syndrome].

Objective To observe the clinical effect of Xinfeng Capsule (XFC) in treating Sjögren's syndrome (SS) and its effect on coagulation parameters, peripheral blood cytokines, and NF- kappa B signaling pathway protein. Methods Totally 58 SS patients were assigned to the treatment group and the control group according to random digit table, 29 cases in each group. Patients in the treat- ment group took XFC, 3 pills each time, 3 times per day. Those in the control group took Hydroxychloro- quine ( HCQ) Sulfate, 0. 1 g per tablet, 2 tablets each time, twice per day. Three months consisted of one therapeutic course and one course for all. Another 20 healthy subjects were recruited as a normal control group. Coagulation parameters (APTT, PT, FIB,TT, D-D) were detected using automatic coagulation an- alyzer in the two groups before and after treatment as well as in the healthy control group. The expres- sions of peripheral blood cytokines (IL-1ß, TNF-α, IL-10) and NF-κB signaling pathway proteins (P65, P50, IκBα) were detected before and after treatment using ELISA. Erythrocyte sedimentation rate (ESR) was determined using Westergren method before and after treatment. High sensitivity C-reactive protein (hs-CRP) level was determined using automatic biochemical analyzer before and after treatment. Results Totally 44 SS patients had abnormal coagulation parameters, accounting for 75. 9% of the total. Com- pared with the healthy control group, the levels of D-D, FIB, IL-1 , TNF-α,P50, P65, IκBα, hs-CRP, and ESR increased (P <0.05, P <0. 01), and the IL-10 level decreased in SS patient groups (P <0.01). Spearman correlation analysis showed that coagulation parameters were significantly correlated with cy- tokines, NF-κB signal transduction pathway, and inflammation indices (P <0. 01 , P <0. 05). After drug in- tervention blood coagulation parameters and laboratory indices were partially improved in the two groups. The effective rate and the total effective rate were 59% and 86% in the treatment group, obviously higher than those of the control group with statistical difference (38% and 72%; P <0. 05). Besides, the treat- ment group was superior to the control group in reducing the levels of FIB, D-D, P50, P65, ESR, and hs- CRP, down-regulating levels of TNF-α and IL-1 ß, as well as up-regulating the expression of IL-10 (P < 0. 05, P <0. 01). Conclusions There is a hypercoagulable state in SS patients, which is related to abnor- mal activation of cytokines/NF-kappa B signaling pathway. XFC could effectively improve the hypercoagulative state of SS patients. Its mechanism might be related to inhibiting cytokines/NF-κB signaling pathway.

[Effect of Xinfeng Capsule in Improving Blood Stasis State of OA Patients].

Objective To observe the relationship between blood stasis state and activation of nuclear factor-κ-gene binding (NF-κB) signaling pathway/miRNA-146 in osteoarthritis (OA) patients and the effect of Xinfeng Capsule (XFC) on them. Methods Totally 70 OA patients were assigned to the treatment group and the control group according to random number table, 35 in each group. Patients in the treatment group took XFC, 3 pills each time, 3 times per day, while those in the control group took Glucosamine, 1 pill each time, 3 times per day. The therapeutic course for all was 3 months. Serum con- tents of p50, p65, inhibitor of nuclear factor κB O (IκBα) , nuclear factor kappa B activator 1 (Act1) , TGF-ß-activated kinase 1 (TAK1) , IL-1, IL-17, IL-10, and thromboxane A2(TXA2) , prostacycline (PGI2) were detected by ELISA. mRNA levels of Act1 , p65, p50, and TAK1 were detected using fluorescent quantitative PCR. The protein levels of p50 and p65 were detected using Western blot. The level of miRNA- 146 was detected using in one-step PCR. Results (1) Compared with pre-treatment in the same group, the levels of blood stasis score, platelets (PLT) , D-dimer (D-D) , TXA2, IL-1, IL-17, high-sensitivity C- reactive protein (hs-CRP), and erythrocyte sedimentation rate (ESR) all decreased; mRNA levels of p50, p65, Act1, and TAK1 were lowered; protein expressions of p50 and p65 decreased; serum levels of miRNA-146 decreased; activated partial thromboplastin time (APTT) , prothrombin time ( PT) , prosta- glandin 2 (PGI2), IL-10 increased in the two groups after treatment with statistical difference (P <0. 05, P <0. 01). Compared with the control group, the levels of blood stasis score, PLT, FIB,TXA2, IL-17, hs- CRP, and ESR were lowered; mRNA expressions of p65 and TAK1 were lowered; protein expressions of p50 decreased; levels of PT and PGI2 increased in the treatment group after treatment (P <0. 05, P < 0.01). Conclusion XFC could regulate the immunity and restore the equilibrium of cytokine network, and protect vascular endothelial cells possibly by up-regulating miRNA-146 expression and inhibiting acti- vation of NF-κB signaling pathway, thus improving blood stasis state of OA.