[Mechanism of Xinfeng Capsules improving rheumatoid arthritis based on CD19~+B cells regulating FAK/CAPN/PI3K pathway].

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)-1ß，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-1ß，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.

[Evaluation of the effect of 3D printed HAP-GEL scaffold combined with BMSCs and HUVECs in repairing rabbit skull defect].

PURPOSE: To qualitatively analyze the effect of 3D printed hydroxyapatite-gel (HAP-GEL) scaffold combined with bone marrow mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) in repairing rabbit skull defect. METHODS: The third generation BMSCs and HUVECs were co-cultured with 3D printed HAP-GEL scaffold to construct tissue engineering bone. The rabbit model of skull defect was established and randomly divided into 4 groups. HAP-GEL stent, HAP-GEL stent + BMSCs and HUVECs cells were implanted respectively, and positive control (autologous bone tissue) and blank control were set up. Twelve weeks after operation, X-ray, cone-beam CT (CBCT) scan and H-E staining were performed to observe and analyze the changes of bone defect qualitatively. RESULTS：Twelve weeks after operation, imaging examination (X-ray and CBCT) showed that there was still obvious circular transmission in the blank control group, and the density was increased and the defect boundary was blurred in both HAP-GEL stent combined cell group and HAP-GEL group, among which the bone was continuous and the bone mineral density was the highest in HAP-GEL stent composite cell group, which was close to normal tissue. The results of H-E staining at twelve weeks showed that compared with the blank control group and the HAP-GEL group, the defect area of the HAP-GEL composite group was filled with new bone and bone-like tissue, the scaffold material was degraded and there was new bone formation inside the scaffold, and the bone repair effect was good, and the osteogenic effect was similar to that of the positive control group. CONCLUSIONS: 3D printed HAP-GEL scaffold + BMSCs + HUVECs cell complex has good osteogenic ability and biocompatibility with a good effect on repairing rabbit skull defect.

Xinfeng capsule for the treatment of rheumatoid arthritis patients with decreased pulmonary function--a randomized controlled clinical trial.

OBJECTIVE: To determine the effectiveness and safety of Xinfeng Capsules (XFC) for the treatment of rheumatoid arthritis (RA) patients with decreased pulmonary function. METHODS: This was a randomized controlled clinical trial of 80 RA patients. Participants were assigned to the trial group (40 cases) and the control group (40 cases) by block randomization. The trial group was treated with XFC, three pills each time three times daily for 2 months. The control group was treated with tripterygium glycoside (TPT), two pills each time three times daily for 2 months. Both groups were followed up after 2 months. The clinical effects, changes in joint and pulmonary function, and quality of life before and after treatment were observed; safety indices were also evaluated. RESULTS: Pain, swelling, tenderness, and duration of morning stiffness of joints were obviously decreased after treatment in both the trial and the control groups compared with baseline (P<0.01). Compared with before treatment, hand grip strength increased significantly after treatment in the trial group (P=0.0000); pulmonary function parameters such as forced expiratory volume in the first second of expiration/forced vital capacity (FEV1/FVC), 50% of the expiratory flow of forced vital capacity (FEF50), carbon monoxide diffusing capacity (DLco) were increased (P<0.01 or P<0.05); measures of quality of life such as role-physical, body pain, vitality and mental health were also improved after treatment in the trial group (all P<0.05). Joint swelling in the trial group decreased compared with the control group (P=0.0043), while hand grip strength was increased after treatment (P=0.0000). The increase in FEF50, DLco, and the dimensions of quality of life such as vitality and mental health were all significantly greater in the trial group than the control group (P<0.05 or P<0.01). CONCLUSIONS: XFC not only relieved joint pain in RA patients, but also significantly improved the ventilation and diffusion function of the lungs. Therefore, XFC could improve the whole body function and enhance the quality of life of RA patients.

Expression of MMP-1/PAR-1 and patterns of invasion in oral squamous cell carcinoma as potential prognostic markers.

BACKGROUND: Matrix metalloproteinase (MMP)-1 degrades type I collagen of the extracellular matrix and also activates protease activated receptor (PAR)-1 to induce angiogenesis. The aims of this study were to evaluate microvessel density (MVD) and the expression of PAR-1 and MMP-1 in oral squamous cell carcinoma (SCC) specimens with different patterns of invasion (POI) and to evaluate their association with clinical outcomes. METHODS: Seventy-four surgically obtained oral SCC samples were classified by POI according to hematoxylin-eosin staining. MVD and the localization and intensity of PAR-1 and MMP-1 expression were detected by immunohistochemistry. RESULTS: Of the 74 oral SCC samples, 18, 5, 34, and 17 showed type I, II, III, and IV POI, respectively. MVD and expression levels of MMP-1 and PAR-1 differed between POI types I-II and POI types III-IV. Patients with low tumor expression of MMP-1 and PAR-1 and low MVD had a longer survival time than those with high tumor expression of MMP-1 and PAR-1. Moreover, the survival time of patients with POI types III-IV was shorter than that of patients with POI types I-II. CONCLUSION: POI combined with expression levels of MMP-1 and PAR-1 may be a valuable tool for assessing the clinical prognosis of patients with oral SCC.

Sonic hedgehog signaling may promote invasion and metastasis of oral squamous cell carcinoma by activating MMP-9 and E-cadherin expression.

The aim of this study was to investigate sonic hedgehog (SHH) signaling pathway components (Shh and Gli-1), E-cadherin, and MMP-9 expression in human oral squamous cell carcinoma (OSCC) and to evaluate their role in prognosis. Expression of Shh, Gli-1, and MMP-9 was significantly upregulated in 74 OSCC samples compared with non-cancerous tissue samples (Shh IOD: 162.44 ± 29.35 and 608.82 ± 170.99; Gli-1 IOD: 203.50 ± 71.57 and 831.11 ± 242.352; MMP-9 IOD: 196.69 ± 64.48 and 721.64 ± 197.99 in non-cancerous and tumor tissues, respectively, P < 0.01), whereas E-cadherin expression was downregulated (E-cadherin IOD: 1,006.19 ± 230.42 and 442.20 ± 156.11; in non-cancerous and tumor tissues, respectively, P < 0.01). Highly expressed proteins were associated with lymph node metastasis; moreover, overexpression of Gli-1 was related to tumor recurrence and cancer clinical staging. Spearman's analysis indicated that the expression of Gli-1 and MMP-9 was positively correlated, whereas expression of Shh/Gli-1 and E-cadherin was negatively correlated. Kaplan-Meier results revealed that patients with low Shh, Gli-1, and MMP-9 expression survived longer than those with high expression. In contrast, low E-cadherin expression was associated with poor prognosis (P < 0.01). In conclusions, transcription factor Gli-1 of the SHH signaling pathway may be an important mediator of invasion and metastasis of OSCC through induced expression of MMP-9 and E-cadherin and may serve as a new prognostic marker.

Expression of αvß6 integrin and collagen fibre in oral squamous cell carcinoma: association with clinical outcomes and prognostic implications.

BACKGROUND: This study aims to investigate the expression and significance of the αvß6 integrin, collagen fibres and matrix metalloproteinases (MMP)-3 in oral squamous cell carcinoma (OSCC) and to analyse the possible regulatory relationships between αvß6, collagen fibres and MMP-3. MATERIALS AND METHODS: A series of 80 patients (mean age 56.4 years) diagnosed with OSCC were enrolled. Associations between αvß6, MMP-3, collagen fibre expression levels and clinicopathological parameters were evaluated using the Fisher exact test. Survival analysis was performed with Kaplan-Meier curves. Spearman rank correlation was used to analyse interactions between αvß6, MMP-3 and collagen fibres. RESULTS: αvß6 and MMP-3 were strongly expressed in human OSCC, especially at the peripheral borders of invasive tumour islands, and collagen fibres were generally disrupted and degraded in the same areas. The expression intensity of αvß6 was associated with the differentiation state of cells. ß6 mRNA was expressed in almost all cancer cells. In carcinomas, αvß6 and MMP-3 expression were correlated with the distribution of collagen fibres. CONCLUSIONS: Tumour cells highly expressing αvß6 have a strong capability for invasion and migration, due to concomitant protease production and the destruction and remodelling of collagen fibres. Increased αvß6 integrin and MMP-3 expression and collagen fibre changes in human OSCCs are related to unfavourable clinical prognostic factors and decreased survival.

[Treatment of rheumatoid arthritis by xinfeng capsule: an efficacy observation].

OBJECTIVE: To observe the curative effect of Xinfeng Capsule (XC) in treatment of rheumatoid arthritis (RA). METHODS: Recruited were 80 active RA patients, who were randomly assigned to the normal control group and the treatment group, 40 in each group. All patients received the same routine anti-rheumatic treatment: Methotrexate 10 mg per week; Diclofenac 50 mg when pain was obvious, twice daily. Patients in the treatment group took XC 3 tablets each time, thrice daily. All treatment lasted for 12 consecutive weeks. Serum iron (SI), serum ferritin (SF), transferrin (TRF); and RA disease activity index (DAS-28) were detected in all patients. RESULTS: XC could improve HAQ, DAS-28, hypersensitive C reactive protein (hs-CRP), prostaglandins A (PGA), erythrocyte sedimentation rate (ESR), number of swelling joints, number of tender joints, and morning stiffness time in acute RA patients, showing statistical difference when compared with those of the control group (P < 0.01, P < 0.05). Compared with the control group, SI, SF, DAS-28, and TRF significantly decreased in the treatment group (P < 0.05). CONCLUSION: XC could improve DAS-28, and SI reserve in patients with active RA, and lower DAS-28 related indicators.

Changes in the expression of MMP2, MMP9, and ColIV in stromal cells in oral squamous tongue cell carcinoma: relationships and prognostic implications.

BACKGROUND: Type IV collagen (ColIV) is the most important scaffold for the basement membrane (BM) proteins, and plays an important role in regulating and limiting tumour invasion and metastasis. METHODS: Here, we observed the changes in morphology and distribution of type IV collagen (ColIV) in the basement membrane (BM) surrounding nests of carcinoma in 48 patients with oral tongue squamous cell (OTSCC). We examined the correlation between the expressions of ColIV, MMP-2 and MMP-9 and the prognosis of OTSCC patients. The intensity and patterns of expression were assessed immunohistochemically using anti-human mouse monoclonal MMP-2, MMP-9 and Col IV antibodies. Statistical analyses were performed to determine the prognostic correlations of ColIV, MMP-2, and MMP-9 levels. RESULTS: MMP-2 and MMP-9 expressions in OTSCC were higher than those in normal oral mucosa and dysplastic oral mucosa group(MMP-2 iOD: 66.40 ± 24.20, 134.69 ± 37.08, and 357.79 ± 116.78; MMP-9 iOD: 88.05 ± 23.85, 307.13 ± 93.22, and 791.31 ± 260.52; in normal, dysplastic oral mucosa, and tumour tissues, respectively, P < 0.01); however, ColIV immunoreactivity was lower (ColIV iOD: 406.87 ± 62.95, 247.83 ± 42.30, and 151.92 ± 38.17 in normal, dysplastic oral mucosa, and tumour tissues, respectively, P < 0.01). High tumour and stromal MMP-2 and MMP-9 expression was significantly associated with positive lymph node status. Col IV expression was associated with positive lymph node status (P < 0.05), and have negatively correlated with the expression of MMP-2 and MMP-9. Overall survival was significantly shorter in patients with high tumour and stromal MMP-2 and MMP-9 expression, and tended to be shorter in patients with low ColIV expression. CONCLUSIONS: Degradation of ColIV was closely related to increased MMP-2 and MMP-9 expression; MMP-9 have more important function than MMP-2 during the cancer development. Monitoring changes in the expression of ColIV, MMP-2, and MMP-9 may be a useful technique for assessing prognoses in OTSCC patients.

[Changes of behavior and brain amino acids of rats with adjuvant arthritis and effects of Xinfeng Capsule on them].

OBJECTIVE: To observe the changes of behavior and amino acids in brain tissue of rats with adjuvant arthritis, and effects of Xinfeng Capsule (XFC) on them. METHODS: AA model was induced by complete Freund's adjuvant in rats. Five groups including the normal control group, the model group, the XFC group, the methotrexate (MTX) group and the Tripterygium wilfordii polysaccharide (TPT) group were set up. The changes of behavior, amino acids in brain tissue, swelling degree of joints, and arthritis index (AI) were observed and the correlation analysis on these indexes was performed. RESULTS: Compared with those in the normal control, the frequency of independent activity obviously decreased in the model rats, accompanied with increased number of step-down errors, shortened step down latency (SDL) and prolonged escape latency (EL), also the level of gamma-aminobutyric acid (GABA) in brain tissue elevated and glutamic acid (GLU)/GABA reduced (P <0.05). Compared with those in the model group, all the above indexes were improved significantly in the XFC group after treatment (P <0.05), while they changed insignificantly in the MTX and TPT groups (P> 0.05), the degree of joint swelling and AI were obviously lower in the 3 treated groups (P< 0.05). Correlation analysis showed the frequency of independent activity and SDL were negatively correlated to the swelling degree of joint, and levels of AI, glycine (GLY) and GABA respectively, but positively correlated with GLU/GABA (P<0.05); while EL and number of step-down errors were positively correlated to AI, GLY and GABA respectively, but negatively correlated to GLU/GABA (P < 0.05). CONCLUSION: Immune stress could lead to the changes of rat behavior and amino acids in brain tissue. XFC treatment could improve the behavior of AA rats through regulating the levels of amino acids in brain tissue.