The biological effect of B7-H3 on fibroblast-like synoviocytes in osteoarthritis / 中华风湿病学杂志

Objective:To investigate the effect of B7-H3 gene on the biological function of fibr-oblastlike synoviocytes (FLS) in osteoarthritis (OA).Methods:Synovial tissue of five cases of OA and synovial tissue of 4 normal knee were obtained, and the primary cell lines were isolated and cultured. The expression of B7-H3 in OA synovial tissue and primary OA-FLS were studied by immunohi-stochemistry, real time-poly merase chain reaction (PCR) and FACS. According to sites 996 and 1041 of B7-H3, corresponding siRNA was designed and the expression of B7-H3 in FLS was silenced and down-regulated. The inhibition of B7-H3 and its protein in target cells was determined by Western blot and FACS. The migration and invasion ability of B7-H3 in target cells were analyzed by scratch assay and Transwell assay. CCK8 assay was used to detect cell proliferation ability, and CBA assay was used to detect cytokines and chemokines in cell culture supernatant. GraphPad Prism 8.0 software was used to analyze the experimental data. The normal distribution data was expressed as mean±standard deviation ( Mean± SD). The comparison between data was performed by T test, and P<0.05 was considered statistically significant. Results:The abnormally high expression of B7-H3 in fibro-blast-like synoviocytes of OA was detected. Compared with siNC, si996 and si1041 inhibited the expression of B7-H3 in OA-FLS. In the Transwell migration experiment, the mean cells number of random view in the siNC group, the si996 group, and the si1041 group indicating decreased migration ability of OA-FLS [siNC vs si996 (100.3±3.7) /view vs (48.7±1.2) /view, t=13.24, P<0.001; siNC vs si1041 (100.3±3.7) /view vs (59.7±1.9) /view, t=9.80, P<0.001). In the Transwell invasion experiment, the mean cells number of random view in the siNC group, in the si996 group, and in the si1041 group indicating decreased invasion ability of OA-FLS [siNC vs si996 (127.3±5.6) /view vs (39.7±3.3) /view, t=13.49, P<0.001; siNC vs si1041 (127.3±5.6) /view vs (57.3±1.9) /view, t=11.85, P<0.001]. The secretion of IL-6 [siNC vs si996 (248±21) pg/ml vs (111±12) pg/ml, t=24.08, P=0.002; siNC vs si1041 (248±21) pg/ml vs (46±5) pg/ml, t=13.21, P=0.006], IL-8 [siNC vs si996 (118.1±15.6) pg/ml vs (47.1±5.4) pg/ml, t=6.68, P=0.022; siNC vs si1041 (118.1±15.6) pg/ml vs (10.0±1.3) pg/ml, t=13.08, P=0.006], CXCL8 [siNC vs si996 (178.8±6.4) ng/ml vs (83.2±2.7) ng/ml, t=13.77, P=0.005; siNC vs si1041 (178.8±6.4) ng/ml vs (93.5±2.8) ng/ml, t=12.23, P=0.007] and CCL2 [siNC vs si996 [(184.1±5.1) ng/ml vs (109.4±5.9) ng/ml, t=9.57, P=0.011; siNC vs si1041 (184.1±5.1) ng/ml vs (97.1±1.5) ng/ml, t=16.39, P=0.004] was decreased . Conclusion:B7-H3 may regulate the migration, invasion, cytokine secretion and other biological functions of OA-FLS, providing clues for further study of B7-H3's involvement in the pathogenesis of OA.

Bioinformatics analysis to investigate immune cell infiltration in synovial lesions of rheumatoid arthritis patients / 中华风湿病学杂志

Objective:To explore the immune infiltration cells in rheumatoid arthritis (RA) synovial lesions, and to provide new research directions and therapeutic targets for the pathogenesis and treatment of RA.Methods:The three gene expression data sets GSE77298, GSE55457 and GSE1919 were downloaded from gene expression omnibus (GEO) (http://www.ncbi.nlm.nih.gov/geo), and the data were merged with Perl. The "limma" package was used to adjust batch differences. In R, "CIBERSORT" software was used to obtain the expression matrix of 22 kinds of immune cells corresponding to RA synovial tissue samples and normal synovial tissue samples were analyzed with the three packages of "e1071", "parallel" and "preprocessCore". Perl was used to screen samples with P<0.05 in the immune cell matrix. R's "barplot" function was analyzed by the percentage of 22 immune cells in samples with P<0.05. The "pheatmap" package of R was used to visualize heatmaps, and "corrplot" package was used to draw correlation heatmaps. The "vioplot" package of R was used to draw violin plots of differences via the wilcox test. Results:The results of immune cell infiltration analysis showed that in RA synovial tissue samples and normal synovial tissue samples at P<0.05, B cells naive and natural killer cells resting were under-expressed in RA synovial tissue, and plasma cells, mast cells resting, macrophages M1, B cells memory and T cells regulatory were highly expressed in RA synovial tissue. This study also found that in the same sample, the correlation coefficient between natural killer cells resting and neutrophils ( r=0.91) was the highest, indicating synergistic effect between the two. In the same sample, the correlation coefficient between macrophages M0 and plasma cells ( r=-0.88) was the lowest, indicating antagonistic effect between the two. Conclusion:The immune infiltrating cells in RA synovial lesions discovered in this study provide a certain theoretical basis and research direction for the research on the disease mechanism and treatment of RA.

Lipoma Arborescente: uma causa de sinovite crônica? / Lipoma Arborescens: a cause of chronic synovitis?

O lipoma arborescente é uma causa incomum de lesão intra-articular que se apresenta como aumento de volume articular indolor, lentamente progressivo, que persiste por muitos anos e é acompanhado por derrames articulares intermitentes. O envolvimento de sítios extra-articulares é incomum, mas pode ocorrer em bainhas tendíneas e bursas. A ressonância magnética é o melhor exame para o diagnóstico, embora a biópsia sinovial possa ser necessária em alguns casos. Relatamos três casos com o objetivo de destacar o espectro clínico da doença, as características da imagem e a resposta ao tratamento imunossupressor.

Lipoma arborescens is an uncommon cause of intra-articular masses that presents as slowly progressive painless swelling of the joint, which persists for many years and is accompanied by intermittent effusions. Extra-articular site(s) involvement is unusual, but can occur in tendon sheaths and bursas. Magnetic resonance imaging is the best diagnostic exam, although synovial biopsy may be necessary. We report three cases in order to highlight the clinical spectrum and imaging features of the disease, so that early diagnosis and appropriate treatment can be given.

Effect of (, TLZT) gel preparation on p53 /miR-502-5p /NF-κBp65 in synovial tissue of knee osteoarthritis / 中国骨伤

OBJECTIVE@#To observe effects of (, TLZT) gel preparation on p53, miR-502-5p, NF-κBp65 in synovial tissue of knee osteoarthritis (KOA), and to explore mechanism of TLZT gel preparation in treating KOA.@*METHODS@#Thirthy-six Wistar rats aged 8 weeks and weighed 200 to 220 g (meaned 208 g) were randomly divided into normal group, model group and traditional Chinese medicine (TCM) group, 12 rats in each group. KOA model was established by modified Hulth method. After 4 weeks of modeling, TCM group treated with TLZT gel preparation for external use, 3 times daily for 2 weeks;normal group and model group were fed normally without intervention. After treatment, morphological changes of specimens in each group were observed, changes of miR-502-5p in synovial tissue were detected by qPCR, and contents of p53, NF-κBp65, IL-1β, TNF-α, MMP-13 in synovial tissue were detected by qPCR and Western Blot respectively.@*RESULTS@#(1)Morphological observation of specimens showed that the articular cartilage in model group was hyaline and uneven, the synovial membranes were hypertrophic and proliferative with a large number of inflammatory cells infiltrating, the joint fluid was thicker in texture;the articular cartilage in TCM group was more transparent and smooth, synovial hyperplasia was mild with a small amount of inflammatory cell infiltration, the texture of articular fluid was clear and sparse. (2) Compared with normal group, content of miR-502-5p of synovial tissue in model and TCM group were increased, mRNA and expression of p53 decreased, expression of NF-κBp65, IL-1β, TNF-α, MMP-13 increased. (3)Compared with model group, content of miR-502-5p in synovial tissue of TCM group decreased (<0.05), mRNA and protein expression of p53 increased (<0.05), mRNA and protein expression of NF-κBp65, IL-1β, TNF-α, MMP-13 decreased (<0.05).@*CONCLUSION@#Expression of p53, miR-502 -5p, NF -κBp65 in synovial tissue is closely related to synovial hyperplasia and inflammatory reaction, TLZT gel preparation may reduce proliferation and inflammatory reaction of KOA synovium by regulating the expression of p53, miR- 502-5p, NF-κBp65 in synovial tissues.

Platelet-rich plasma plays a protective role in chondrocytes by regulating synovitis / 中国组织工程研究

BACKGROUND: Studies have confirmed that inflammatory factors secreted by synovitis that accelerate the catabolism of articular cartilage have become the main cause of osteoarthritis. Matrix metalloproteinase 3 and matrix metalloproteinase 13 play a key role in osteoarthritis. OBJECTIVE: To study the effective mechanism of platelet-rich plasma (PRP) in the treatment of osteoarthritis. METHODS: The rat knee synovial cells and chondrocytes were isolated and extracted separately. The blood samples of rats were extracted to prepare PRP preparation. Then, synovial cells were divided into a control group and an E. coli lipopolysaccharide (LPS) treatment group, where synovial cells were stimulated with LPS to create a synovitis model. The synovitis cells were further divided into a PRP treatment group and an untreated group. The cells in each group were cultured for 24 hours. A portion of the medium was taken, in which the levels of interleukin-1β, interleukin-6 and tumor necrosis factor-α was detected using ELISA. The other part of the medium was used to treat chondrocytes. After 48 hours of culture, the changes of type I, II collagens, matrix metalloproteinase 3 and matrix metalloproteinase 13 protein levels in chondrocytes were detected by western blot. The expression levels of matrix metalloproteinase 3 and matrix metalloproteinase 13 mRNA were detected by real-time PCR. RESULTS AND CONCLUSION: Determined by the ELISA, the levels of interleukin-1β, interleukin-6 and tumor necrosis factor-α in the synovial cell culture medium of the LPS treatment group were significantly higher than those of the control group and the PRP treatment group (P < 0.01). The levels of interleukin-1β, interleukin-6 and tumor necrosis factor-α in the medium of PRP treatment group were significantly higher than those in the control group (P < 0.05). Western blot showed that the expression of type I and II collagens in the chondrocytes of the PRP treatment group were significantly higher than those in the control and LPS treatment groups (P < 0.01); the expression of type I and II collagens in the chondrocytes of the LPS treatment group was significantly lower than that in the control group (P < 0.05). The expression levels of matrix metalloproteinase 3 and matrix metalloproteinase 13 proteins in the LPS treatment group were significantly higher than those in the PRP treatment and control groups (P < 0.01). PCR analysis showed that the mRNA expression of matrix metalloproteinase 3 and matrix metalloproteinase 13 in the LPS treatment group was significantly higher than that in the PRP treatment and control groups. The findings of this study reveal that PRP treatment can reduce interleukin-1β, interleukin-6 and tumor necrosis factor-α levels in synovial cells, decrease matrix metalloproteinase 3 and matrix metalloproteinase 13 protein and mRNA levels in chondrocytes, and increase type I and II collagen expression, thereby protecting the cartilage.

Cartílago y gonartrosis / Cartilage and knee osteoarthritis

RESUMEN Fundamento: el cartílago articular es un tejido avascular, aneural y alinfático que desempeña un importante papel en la articulación, su afección más frecuente es la de tipo degenerativa. Objetivo: actualizar los conocimientos sobre el cartílago articular normal, envejecido y con cambios degenerativos. Métodos: la búsqueda y análisis de la información se realizó en un periodo de tres meses (primero de octubre al 31 de diciembre de 2018) y se emplearon las siguientes palabras: cartilage AND osteoarthritis, cartilage AND knee osteoarthritis, cartilage a partir de la información obtenida se realizó una revisión bibliográfica de un total de 164 artículos publicados en las bases de datos PubMed, Hinari, SciELO y Medline mediante el gestor de búsqueda y administrador de referencias EndNote, de ellos se utilizaron 50 citas seleccionadas para realizar la revisión, de ellas 48 de los últimos cinco años. Resultados: se mencionan los aspectos macro y microscópicos del tejido, así como su organización por zonas y áreas. Se describen los cambios que ocurren en el proceso degenerativo a diferentes niveles y en el envejecimiento. Conclusiones: el cartílago articular es la estructura anatómica más afectada en la articulación por el proceso degenerativo. Se encuentra organizado por zonas y áreas, las que se afectan a medida que avanza la enfermedad. El origen de la destrucción del cartílago es enzimático y repercute en las demás estructuras como el tejido sinovial y hueso subcondral. Es importante conocer las diferencias entre el envejecimiento y la afección degenerativa de este tejido.

ABSTRACT Background: articular cartilage is an important avascular, alinphatic and aneural tissue in joints, it is mainly affected by degenerative disease. Objective: to update knowledge about normal, aging and degenerative articular cartilage. Methods: a three months research and analysis were conducted from October 1st, 2018 to December 31th, 2018. Our review included 164 articles published in PubMed, Hinari, SciELO and Medline databases by using EndNote. The words used were cartilage AND osteoarthritis, cartilage AND knee osteoarthritis, cartilage. Fifty citations were selected, 48 of all them within the last five years, were used to write the present paper. Results: macro and microscopic features of articular cartilage were mentioned as well as its organization in zones and areas. Degenerative process was described at different levels, and in aging. Conclusions: articular cartilage is the most affected tissue in osteoarthritis. Cartilage is organized by zones and areas which are affected as degenerative disease progresses. The start point of osteoarthritis is enzymatic and gradually affects synovial tissue and sub-chondral bone. It is important to know the main differences between aging and degenerative cartilages.

Comportamiento de pacientes con plica sinovial de la rodilla / Behavior of patients with knee plica synovialis

RESUMEN Fundamento: la plica sinovial de la rodilla es una de las causas de dolor anterior, se presenta de forma asilada o combinada a otras afecciones intrarticulares. El tratamiento quirúrgico mediante la vía artroscópica es el más usado en la actualidad. Objetivo: evaluar los resultados del tratamiento artroscópico en pacientes con plica sinovial. Métodos: se realizó un estudio observacional analítico con un nivel de evidencia III recomendación C en 181 pacientes con el diagnóstico clínico, imagenológico y artroscópico de plica sinovial en el Hospital Universitario Manuel Ascunce Domenech desde el 1 de enero de 2012 al 31 de enero de 2018. La población de estudio estuvo dada por todos aquellos enfermos en que se confirmó la presencia de plica sinovial patológica mediante la vía artroscópica, edad de 18 años o más y respuesta limitada o nula al tratamiento conservador ambulatorio por más de seis semanas. Resultados: el promedio de edad de los pacientes estudiados fue de 45,5 años, la razón sexo femenino-masculino fue de 1,9 a 1. Predominó la plica mediopatelar. Las lesiones de cartílago grados III/IV fueron las más encontradas, así como la afección monocompartimental patelofemoral. El desbridamiento y la meniscectomía fueron los procedimientos artroscópicos más realizados después de la sección de la plica. La respuesta clínica de los pacientes a los seis meses fue satisfactoria. Conclusiones: el tratamiento artroscópico de la plica sinovial es efectivo, en especial para pacientes con ausencia de otras lesiones intrarticulares.

ABSTRACT Background: knee plica synovialis is a common cause of anterior knee pain; it could show up isolated or associated to other intra-articular conditions. Nowadays, the surgical arthroscopic treatment is the most used way. Objective: the objective of this study was to evaluate the arthroscopic treatment in patients with plica synovialis of the knee. Methods: an analytic and observational study, with a level of evidence 3, recommendation C was performed in 181 patients with the clinical diagnosis of plica synovialis of the knee according to clinical, imaging and arthroscopic criteria in the provincial teaching hospital Manuel Ascunce Domenech in Camaguey city from January 1st, 2012 to January 31th, 2018. The population of the research was the amount of patients who had had the confirmation of having pathologycal plica synovialis through the arthroscopic way, 18 year-old patients and over and also limited or non-response to conservative ambulatory treatment for more than six weeks. Results: the average age was 45,5 years, female-male ratio was 1,9 to 1. The plica synovialis mediopatellaris prevailed. Cartilage lesions levels 3 and 4 were the most found as well as mono-compartiment affecting patellofemoral joint. Debridement and meniscectomy were the most common used arthroscopic procedures after plica section. Clinical results were good at six months. Conclusions: the arthroscopic treatment of plica synovialis is effective, especially in patients with absence of other intrarticular conditions.

Role of TLR4/NF-κB pathway for early change of synovial membrane in knee osteoarthritis rats / 中国骨伤

OBJECTIVE@#To study role of TLR4/NF-κB pathway for early change of synovial membrane in knee osteoarthritis rats.@*METHODS@#Eighteen male SD rats weighted (200±20) g were randomly divided into 2 groups, namely control and model group, and 9 in each group. Knee OA model group was established by using modified Hulth method in model group. Control group was not treated. Synovial tissue and serum was extracted at 4 and 21 d after operation. Expression of CD14, TLR4, IL-1β, TNF-α, ADAMTS-4, MMP-13 were detected by real-time PCR respectively. NF-κB p65 protein was detected by Western-blot; serum concentrations of haluronic acid (HA), N-propeptide of type III procollagen(PIIINP) was detected by Elisa.@*RESULTS@#Expression of CD14, ADAMTS-4, and NF-κB p65 in model group were higher than that of control group at 4 and 21 days after operation, while expression of TLR4, IL-1β, TNF-α and MMP-13 were higher than that of control group at 21 days after operation(<0.01). Concentration of PIIINP and HA in model group were higher than that of control group at 4 days after operation, while there was no significant difference at 21 days after operation.@*CONCLUSIONS@#NF-κB pathway could mediate occurrence of KOA by early activating and triggeringg synovial increasingly secreting inflammatory secretion CD14, TLR4, IL-1β, TNF-α, ADAMTS-4, MMP-13, PIIINP and HA.

miR-124a inhibited cell proliferation, migration and invasion of rheumatoid arthritis synovial fibro-blasts by tar-geting AKT2 gene / 中华风湿病学杂志

Objective@#To investigate the effects of miR-124a on proliferation, migration and invasion in rheumatoid arthritis synovial fibroblasts (RASFs) and the underlying mechanisms.@*Methods@#RASFs were isolated and cultured from synovial tissue, then qRT-PCR was used to detect the levels of AKT2 mRNA and miR-124a in RASFs. Western blot was applied to determin the expression level of AKT2 protein. RASFs were transfected with miR-124a, anti-miR-124a, si-AKT2 or pcDNA-AKT2 to up-regulate or down-regulate the expression level of miR-124a or AKT2 protein. The cells were divided into normal group of normal synovial tissue, control NC group, miR-con group, miR-124a group, si-con group, si-AKT2 group, miR-124a+pcDNA group and miR-124a+pcDNA-AKT2 group. MTT assay was carried out to measure the proliferation of RASFs. Transwell assay was carried out to detect the migration and invasion cell number of RASFs. Dual-luciferase reporter assay system was implemented to verify the relationship between miR-124a and AKT2. Independent sample t-test and one way analysis of variance (ANOVA) test (square deviation) were used for statistical analysis.@*Results@#① Compared with normal group, the expression of miR-124a (0.92±0.19) decreased significantly (t=5.788, P<0.01), AKT2 mRNA (3.15±0.63) increased significantly (t=-3.486, P=0.025), AKT2 protein (2.09±0.64) increased significantly (t=-2.959, P=0.042). ② Ccompared with NC group and miR-con group, miR-124a expression (4.17±0.46) increased significantly (F=131.830, P<0.01), migration cell number (34±6) decreased significantly, invasion cell number (14.5±3.1) decreased significantly (F1=35.788, F2=27.211, P<0.01). ③ Compared with mir-con group (1.02±0.18), WT-AKT2 in miR-124a group showed a significant decrease in its relative activity (0.31±0.11) (t=5.830, P<0.01). ④ Compared with NC group and si-con group, the expression of AKT2 protein (0.97±0.03) in si-AKT2 group decreased significantly (F=128.056, P<0.01), the number of migrating cells (32±4), and the number of invasive cells (18.6±2.2) (F1=-70.082, F2=36.524, P<0.01) were decreased significantly. ⑤ Compared with miR-con group, AKT2 protein expression in miR-124a group decreased significantly (0.21±0.03); compared with miR-124a+pcDNA group, AKT2 protein expression in miR-124a+pcDNA-AKT2 group was increased significantly (F=52.487, P<0.01). ⑥ Compared with miR-con group, the number of RASFs migrating cells (30±5) and invasive cells (12.5±1.8) in miR-124a group were significantly decreased; compared with miR-124a+pcDNA group, the number of RASFs migrating cells (71±4) and invasive cells (26.4±4.5) in miR-124a+pcDNA-AKT2 group were significantly increased (F1=30.957, F2=49.960, P<0.01).@*Conclusion@#MiR-124a can inhibite the proliferation, migration and invasion of RASFs by targeting AKT2 gene. MiR-124a is expected as a molecular target for diagnosis and treatment of rheumatoid arthritis.

The comparison of two different intraarticular injections using a sonographic anterolateral approach in patients with osteoarthritic knee

BACKGROUND: The intraarticular (IA) injection has become popular for the management of the osteoarthritic knee without an effusion. The success rate of IA injection would be better if it was able to be visually confirmed. We hypothesized that an anterolateral approach, which targets the synovial membrane of the lateral condyle using ultrasound, would provide an equivalent alternative to the anterolateral approach, targeting the synovial membrane of the medial condyle for IA injection of the knee. METHODS: A total of 96 knees with osteoarthritis were randomized placed into the two groups, which were group I (anterolateral approach to the medial condyle) and group II (anterolateral approach to the lateral condyle). The primary outcome was to compare the success rate of the two methods of IA injection. The required length of the needle for injection was also measured and compared. Pain intensity was assessed using the Numeric Rating Scale in order to evaluate the success of injection. RESULTS: There were no significant differences in the success rate between both groups. The success rate of group I and group II were 87.8% (95%, CI 78.7–97.0) and 91.5% (95%, CI 83.6–99.5), respectively (P = 0.549). The needle depth was 5.0 ± 0.8 (3.0 to 6.1 cm) in group I, and 3.0 ± 0.8 (1.5 to 5 cm) in group II (P < 0.001). CONCLUSIONS: The anterolateral approach to the lateral femoral condyle, using ultrasound, is an alternative method to the approach targeting the medial femoral condyle, using shorter needle.

Correlation of synovectomy with blood loss and knee joint function in primary total knee arthroplasty / 中国组织工程研究

BACKGROUND: In recent years, knee arthroplasty is a relatively mature method for the treatment of various advanced osteoarthritis, and has good clinical efficacy. However, there is still a lot of controversy about whether to retain the synovial membrane. OBJECTIVE: To analyze the effect of synovectomy in primary total knee arthroplasty on the blood loss and knee joint function in the patients with knee osteoarthritis. METHODS: Sixty patents with knee osteoarthritis were included, involving 25 males and 35 females, with an age of 55-70 years, and were then randomly divided into two groups (n=30 per group). The patients underwent total knee arthroplasty with (experimental group) or without (control group) synovectomy. The operation time, dominant, hidden and total bold loss, and hospitalization time were recorded. The Visual Analogue Scale scores at baseline, postoperative 3 days, 4 and 12 months were detected; the Hospital for Special Surgery scores at baseline, postoperative 4 and 12 months were determined. RESULTS AND CONCLUSION: (1) The operation time, as well as the dominant, hidden and total bold loss in the experimental group were significantly more than those in the control group (P < 0.05). There was no significant difference in hospitalization time between two groups (P > 0.05). (2) The Visual Analogue Scale scores at beseline and postoperative different time points showed no significant difference between two groups, and the postoperative scores were significantly improved in both groups (P < 0.05). (3) No significant difference was found in the Hospital for Special Surgery scores between two groups at baseline and postoperative different time points, and the postoperative scores in both groups were significantly higher than those at baseline (P < 0.05). (4) These results imply that the removal of synovial membrane during total knee arthroplasty can induce large amount of blood loss, and has no effect on the knee function in the patients with knee osteoarthritis.

Experimental study of 99Tcm-3PRGD2 imaging on the angiogenesis of rheumatoid arthritis / 中华核医学与分子影像杂志

Objective@#To observe the uptake of 99Tcm-3 polyethylene glycol Arg-Gly-Asp dimer (3PRGD2) in rat models of rheumatoid arthritis (RA), in order to provide theoretical foundation for early diagnosis of RA.@*Methods@#The healthy female SD rats were divided into collagen induced arthritis (CIA) group (n=100), osteoarthritis (OA) group (n=20) and control group (n=20). Bovine collagen type Ⅱ emulsion was used for arthritis induction to establish CIA models. OA models were established by injection of L-cysteine and papain. Gamma imaging was performed before and 15 d, 30 d after the model establishment. The mediastinum was selected as non-target (NT) area and the target (T)/NT ratios were calculated. The serum levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-α (TNF-α), αVβ3 before and after the model establishment were measured. Pathological and immunohistochemical detection were performed. One-way analysis of variance, Pearson and Spearman correlation analyses were used to analyze the data.@*Results@#There were 58 CIA models successfully established. Before the model establishment, T/NT ratios of CIA group, OA group and control group were 0.215±0.049, 0.210±0.050, 0.209±0.051, respectively (F=0.093, P>0.05). The T/NT ratios of the above three groups were 0.405±0.230, 0.223±0.045, 0.211±0.049 (F=12.601, P<0.05) 15 d post-model establishment, and those were 0.572±0.182, 0.238±0.045, 0.212±0.055 (F=65.147, P<0.05) 30 d post-model establishment. T/NT ratios of CIA group were positively correlated with the levels of serum VEGF, TNF-α, αVβ3, pathological score and the levels of immunohistochemical markers (VEGF, TNF-α, αVβ3, CD31, CD34; r values: 0.391-0.721, rs values: 0.365-0.669, all P<0.05).@*Conclusion@#99Tcm-3PRGD2 can be specifically up-taken by RA lesions in rat models, thus has the potential in early synovial neovascularization imaging of RA.

Establishment of rat models with altered anterior guidance and observation of histological changes in temporomandibular joint synovium / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aims to establish a stable animal model with altered anterior guidance and investigate histological changes on temporomandibular joint (TMJ) synovium in rats.</p><p><b>METHODS</b>Thirty-two rats were divided into four groups, namely, control group, anterior-guidance-deficient group (T1), and anterior guidance inclined up to 15 degree (T2) and 30 degree (T3) groups. Metal crowns that add 15 and 30 degrees to the palatal side were cemented to the incisors of maxilla to increase the inclination of anterior guidance. The low-speed diamond wheel shorts for both upper and lower incisors of rats were applied to avoid contact between anterior teeth in all direction, thus establishing an anterior guidance deletion model. TMJ joints on one side of two animals in each group were harvested after 3, 7, 14, and 28 days. Pathological changes were investigated in the TMJ synovium using hematoxylin-eosin (HE) staining.</p><p><b>RESULTS</b>The weight of T1 decreased briefly at first, and then increased slightly (P<0.05). The weight of T3 obviously reduced and then slowly increased, but remained below the initial weight (P<0.05). T3 manifested obvious synovial lining proliferation and vascular dilation (P<0.05).</p><p><b>CONCLUSIONS</b>The rat model of anterior guidance inclined up to 30 degree could imitate the pathological features of TMJ synovium with excessive increase in anterior guidance. A higher inclination of anterior guidance might cause more damage of TMJ synovium. The effect of anterior guidance deletion on TMJ synovium is yet to be determined.
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Whole-genome expression analysis of synovial fibroblasts in rheumatoid arthritis / 中华骨科杂志

Objective To identify the crucial gene implicated in rheumatoid arthritis (RA) pathogenesis by comparing microarray-based gene expression profiles of synovial fibroblast in arthritis patients and that in control.Methods The public datasets were obtained from NCBI GEO and EBI ArrayExpress.The qualified microarray-based gene expression profiles were integrated and normalized using the method implemented in GeneSpring software.Furthermore,the differentially expressed genes (DEGs) were identified using significance analysis of microarrays (SAM) method.The online tool DAVID and STRING were applied to conduct the enrichment analysis and gene product interaction analysis respectively.Results There were two datasets that were qualified and analyzed in the present study.A total of 336 significant DEGs were identified by comparing the whole-genome gene expression profiles from synovial fibroblast of RA patients and control group.Among these DEGs,261 were significantly downregulated and 75 upregulated.About 13.6％ of the downregulated genes were associated with extracellular matrix degradation.The COL9A3 and COL4A5,indispensable component of hyaline cartilage and basement membrane respectively,were significantly downregulated,as well as genes in WNT family,including WNT2,WNT11,and WNT16.In contrast,matrix metalloproteinase 13 (MMP13) was found to be significantly upregulated in RA patients.MMP13 is a matrix metallopeptidase that degrade extracellular matrix and hyaline cartilage,and it could possibly interact with other proteins to regulate morphogenesis.Conclusion Molecular mechanisms underlying RA pathogenesis were investigated by analyzing the public datasets.A few genes that associated with extracellular matrix degradation,construction and regulation,including MMP13,WNT2,WNT11,WNT16,COL9A3 and COL4A5,could be regarded as therapeutic targets in RA treatment.

A preliminary study on the promotion of angiogenesis by NETs in rheumatoid arthritis / 天津医药

Objective To investigate the effect of neutrophil extracellular traps (NETs) on inflammation of rheumatoid arthritis (RA), especially angiogenesis. Methods The presence of NETs in synovial tissues of RA and osteoarthritis (OA) was observed by immunofluorescence assay. Neutrophils were isolated from peripheral blood of health volunteers. Neutrophils were cultured in vitro, the formation of NETs was observed. NETs were extracted as a stimulating agent. The effects of NETs on the proliferation of human umbilical vein endothelial cells (HUVECs) and synovial fibroblasts (RAFLS) were evaluated by MTT, and which were classified into two groups: HUVECs group and RAFLS group, with the following treatment: control and NETs (0.28 mg/L). Wound repair assay was employed to evaluate the effect on the cell migration stimulated with NETs. The experiment was divided into three groups:control, VEGF (40μg/L VEGF) and NETs (0.28 mg/L NETs). Results (1) Compared with OA, NETs were found more in the synovial tissue of RA. (2) NETs formation was induced by stimulator in vitro. The concentration of extracted NETs-DNA was 27.8 mg/L. (3) MTT assay showed that compared with the control groups, low concentration of NETs (0.28 mg/L) promoted the proliferation of HUVECs (0.499 ± 0.011 vs. 0.393 ± 0.009, P<0.05) and RAFLS (0.266 ± 0.007 vs. 0.192 ± 0.007, P<0.05). (4) It was showed that a significant wound closure induced by low concentration of NETs (0.28 mg/L) was found compared with control. Conclusion Our present study suggests that NETs are found more in the synovial tissue of RA, and low concentration of NETs can promote angiogenesis in RA.

Preliminary study of contrast-enhanced ultrasonography on the efficacy of knee synovial lesions in rheumatoid arthritis / 中华超声影像学杂志

Objective To investigate the effect of contrast-enhanced ultrasonography (CEUS) on the treatment of knee synovial lesions in rheumatoid arthritis (RA).Methods The results of routine ultrasonography (US) and CEUS were observed in 37 patients with RA.Among them 26 knees were underwent review after treatment.The results before and after treatment were compared.Results Routine US showed that the synovial thickness of patella,medial and lateral condylar and the depth of suprapatellar bursa effusion in 37 knee joints were (0.47 ± 0.26)cm,(0.31 ± 0.15)cm,(0.36 ± 0.21)cm and (0.72 ± 0.42)cm before treatment,and (0.36± 0.16)cm,(0.28 ± 0.17)cm,(0.30 ± 0.19)cm and (0.41 ± 0.19)cm in 26 knee joints after treatment,respectively,the differences were statistically significant(P ＜0.05).The synovial blood flow classification of patella,medial and the lateral condyle in the 26 knee joints had difference between before and after treatment (P ＜0.05).CEUS showed that the peak intensity decreased,the area under the curve reduced,the time from peak to one half decreased,the wash in slope decreased and the time to peak prolonged in synovial after treatment,the differences of the parameters between before and after treatment were statistically significant(P ＜0.05).The area of synovial had some influence on the CEUS parameters and could improve the reliability of the evaluation to CEUS for treatment.Conclusions CEUS is an objective method to evaluate the efficacy of RA,which provides a reliable basis for clinical treatment of RA.

Analysis of factors related to the number of mesenchymal stem cells derived from synovial fluid of the temporomandibular joint / 中华口腔医学杂志

Objective@#To analyze related factors on the number of mesenchymal stem cells in the synovial fluid of the temporomandibular joint (TMJ) and provide an research basis for understanding of the source and biological role of mesenchymal stem cells derived from synovial fluid in TMJ.@*Methods@#One hundred and twenty-two synovial fluid samples from 91 temporomandibular disorders (TMD) patients who visited in Department of TMJ Center, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University from March 2013 to December 2013 were collected in this study, and 6 TMJ synovial fluid samples from 6 normal volunteers who were studying in the North Campus of Sun Yat-sen University were also collected, so did their clinical information. Then the relation between the number of mesenchymal stem cells derived from synovial fluid and the health status of the joints, age of donor, disc perforation, condylar bony destruction, blood containing and visual analogue scale score of pain were investigated using Mann-Whitney U test and Spearman rank correlation test.@*Results@#The number of mesenchymal stem cells derived from synovial fluid had no significant relation with visual analogue scale score of pain (r=0.041, P=0.672), blood containing (P=0.063), condylar bony destruction (P= 0.371). Linear correlation between the number of mesenchymal stem cells derived from synovial fluid and age of donor was very week (r=0.186, P=0.043). The number of mesenchymal stem cells up-regulated when the joint was in a disease state (P=0.001). The disc perforation group had more mesenchymal stem cells in synovial fluid than without disc perforation group (P=0.042).@*Conclusions@#The number of mesenchymal stem cells derived from synovial fluid in TMJ has no correlation with peripheral blood circulation and condylar bony destruction, while has close relation with soft tissue structure damage of the joint.

Papel da exposição à Hidroquinona na artrite reumatoide experimental induzida pelo colágeno / Role of Hydroquinone exposure on experimental collagen-induced arthritis

Artrite reumatoide (AR) é uma doença autoimune, que causa inflamação crônica nas membranas sinoviais de diversas articulações. O modelo experimenal de artrite induzida pelo colágeno (AIC) é empregado para investigar os mecanismos da AR e para identificar potenciais agentes terapêuticos. Embora a etiologia da AR ainda seja desconhecida, há evidências que a AR se desenvolve em indivíduos predispostos geneticamente, após exposição a fatores ambientais, como o tabagismo, que se destaca como maior fator de risco para indução da AR e para o agravamento em pacientes com AR já estabelecida. Porém, o mecanismo efetivo da ação dos diversos componentes do cigarros ainda precisa ser elucidado. A Hidroquinona (HQ) é um composto fenólico, encontrada em concentração elevada no cigarro, com maior ativade pró-oxidativa, além de ser produto da biotransformação do benzeno, também encontrado no cigarro. Neste caso, a HQ é responsável pela imunotoxicidade e mielotoxicidade do benzeno. Devido a alta exposição de fumantes à HQ e a associação do tabagismo com a AR, investigamos se a exposição à HQ teria participação no desenvolvimento da AIC em ratos Wistar. Para tanto, animais foram expostos à HQ em diferentes protocolos experimentais, a saber: A - por 35 dias consecutivos, durante fase de indução e desenvolvimento da artrite; B - por 14 dias consecutivos, até a segunda injeção de colágeno, na fase de sensibilização e indução da AIC; C - por 7 dias consecutivos, do 29º ao 35º dia, na fase posterior ao desenvolvimento da AIC. Os resultados obtidos mostraram que a HQ agravou a AR nos 3 grupos experimentais, aumentando os parâmetros clínicos, o número de células no líquido sinovial, a inflamação nas sinóvias, caracterizada por maior influxo de neutrófilos, proliferação de sinoviócitos (histologia por HE e imunohistoquímica), aumento nos níveis de IL-6 e IL-1ß (ELISA) no líquido sinovial e rearranjo do colágeno na sinóvia (microscopia por segundo harmônico). No entanto, os efeitos mais acentuados foram observados em animais dos grupos A e C, que também tiveram perda de peso significativa. Ademais, exposição à HQ, nos 3 grupos experimentais, causou expressão aumentada do receptor aril hidrocarboneto (AhR), um receptor ativado por xenobióticos durante a AR, e aumento nos níveis do fator de transcrição ROR e de IL-17 na sinóvia. Como AhR/ROR/IL-17 em linfócitos e neutrófilos é uma via importante na gênese da AR, ensaios in vitro foram realizados para elucidar o papel da HQ nesta via. A incubação com HQ in vitro de esplenócitos de animais naive elevou a expressão de AhR e de secreção de IL-17 (por citometria de fluxo), as quais foram bloqueadas pelo antagonista de AhR (α-naftoflavona). Em conjunto, os resultados obtidos nos permitem concluir que a HQ, como um importante componente do cigarro agrava a CIA em ratos, e a ativação via AhR/IL-17 é um possível mecanismo da patogênese da artrite

Rheumatoid arthritis (RA) is an autoimmune disease that causes chronic inflammation in the joint synovial membranes. The experimental model of collagen-induced arthritis (CIA) is used to investigate the involved mechanisms in RA and to identify novel therapeutic agents. The genesis of RA is multifactorial, involving interplay of genetic and environmental factors and smoking is the trigger factor in the development or RA and worsens the pre-existing RA but the mechanisms undlerlying are yet to be elucidated. Hydroquinone (HQ) is a phenolic compound, found in high concentrations in cigarette, where HQ is the major oxidative component. Moreover, HQ is benzene metabolite, which is also found in cigarette smoke, being responsible for the myelotoxicity and immunotoxicity detected during benzene exposure. Due to this association, we aimed to investigate the role of HQ exposure on CIA development in Wistar rats and the involved mechanisms. Animals were exposed to HQ according to different protocols: A - during 35 consecutive days, during the sensitization and devolpment phases of the disease; B - during 14 consecutive days, until the second injection of collagen, during the sensitization phase; C - during 7 consecutive days, from day 29 to 35, after the development phase of CIA. The results showed that HQ worsened the RA in the 3 experimental protocols, HQ elevated the clinical parameters of CIA development, increased inflammation in the synovial membrane, characterized by increased influx of neutrophis, synoviocytes proliferation (visualized by Immunohistochemistry and Histology analysis), augmented the levels of IL-6 and IL-1ß in the synovial fluid (ELISA assay) and led to intense collagen deposition on the synovia. The most pronounced effects where observed in animals from groups A and C, which also had weight body loss. In addition, in the 3 protocols, HQ exposure also increased the expression of AhR receptor, a receptor activated by xenobiotics during RA, and increased the expression of ROR and levels of IL-17 secretion in the synovial membranes. As AhR/ROR/IL-17 in lymphocytes and neutrophils is an important pathway involved in the genesis of RA, in vitro studies have been performed to elucidate the role of HQ exposure in this pathway. The HQ in vitro treatment augmented the expression of AhR and secretion of IL-17 by splenocytes (FACS assay) and the administration of an AhR antagonist (α-naphtoflavone) blocked these effects. Taken together, the results obtained here allow us to conclude that HQ, as an important cigarette component, aggravates CIA in rats, and the activation of AhR/IL-17 pathway is a possible mechanism involved in the RA pathogenesis

Calreticulin-mediated upregulation of anti-apoptotic Bcl-XL and Mcl-1 promotes survival of fibroblast-like synoviocytes in rheumatoid arthritis / 中华风湿病学杂志

Objective Fibroblast-like synoviocytes (FLS) play important roles in the pathogenesis of rheumatoid arthritis (RA).The present study was undertaken to investigate the mechanism of calreticulin (CRT) to promote FLS survival in RA.Methods FLS were isolated by enzymatic digestion of synovial tissue specimens obtained from RA and osteoarthritis (OA) patients and cultured in vitro.The expression of Bcl-XL and Mcl-1 in FLS at mRNA and protein level was detected by quantitative-polymerase chain reaction (q-PCR),Western blotting and immunofluorescence respectively.RA and OA FLS were cultured with different concentrations of recombinant human CRT for 48-72 h,the expression of Bcl-XL and Mcl-1 was detected by q-PCR and Western blotting.The proliferation of RA FLS following CRT stimulation was determined by MTT assay.Results ① Compared with FLS from OA patients (1.00±0.39;1.00±0.46),the anti-apoptotic Bcl-XL and Mcl-l mRNA expression (14.51 ±2.20;12.82±1.80) was significantly higher in the FLS from RA patients (t=10.47,1 1.02;P＜0.01);Western blotting analysis also showed increased protein levels of Bcl-XL and Mcl-1 in RA FLS;Immunofluorescence results showed higher expression of Bcl-XL and Mcl-1 in RA at the single FLS level;② CRT up-regulated the expression of Bcl-XL and Mcl-1 in RA FLS:compared with the control group (0 ng/ml),CRT stimulation at 10 ng/ml and 50 ng/ml increased the levels of Bcl-XL mRNA (1.70±0.28 vs 1.00±0.20,q=4.58,P＜0.05;1.87±0.35 vs 1.00±0.20,q=5.69,P＜O.05) and Mcl-1 mRNA (1.85±0.36 vs 1.00±0.20,q=5.63,P＜0.05;1.72±0.26 vs 1.00±0.20,q=4.77,P＜0.05) in RA FLS,while no significant effects of CRT on Bcl-XL and Mcl-1 mRNA expression were observed in OA FLS (F=1.49,1.60;P＞0.05);Western blotting results showed elevated protein levels of both Bcl-XL and Mcl-1 in RA FLS after CRT treatment at a concentration dependent manner.However,neither Bcl-XL nor Mcl-1 expression was significantly changed in OA FLS.③ MTT assay showed that CRT had no significant effect on the proliferation of RA FLS (F=2.88,P＞ 0.05).Conclusion Our results indicate that CRT-mediated up-regulation of anti-apoptotic Bcl-XL and Mcl-1 may inhibit apoptosis and promote the survival of FLS from RA patients.

Primary culture for synovial fibroblasts in mice / 重庆医学

Objective The primary culture of synovial fibroblasts is a convenient tool to study the pathology and physiology of synovial tissues .An improved method was constructed in this study by C57BL /6 mice to study the mechanism of rheumatoid ar-thritis(RA) .Methods The synovium around the hip joints were collected .Attention should be paid to eliminate the egg-yolk like yellow oval substance in the middle of the synovium .The synovium was transferred into a 1 .5 mL Eppendorf tube containing 0 .5%type Ⅳ collagenase and cut into 1 mm3 blocks or so .The Eppendorf tube was placed in 37 ℃ Constant temperature orbital shaker incubator for 60 min .After digestion ,the tube was placed on the Vortex for a high-speed oscillation for 1 .5 minutes to guarantee the separation of cells .Results Within about 1 week ,the first passage was performed by the trypsin digestion method .On day 10 , the number of synovial macrophages reached the maximum and then decreased gradually .After the third generation (day 15 to 20) , the synovial macrophages generally disappeared .Vimentin was suitable for the immunofluorescence cytochemical staining for the synovial fibroblasts .The cell purity was indicated as > 95% .The cytometric analysis indicated that purity of Vimentin and CD90 .2-labelled cells was over 95% ;the purity of CD54-labelled cells was 80% approximately .Conclusion It is a simple and effective method for primary culture of synovial fibroblasts in mice .