Fractionation of protein components of the human synovial fluid was carried out with paper and disc electrophoresis, and isoelectric focusing. The mean ranges of total protein content of synovial fluid obtained in the thirty patients suffering from nonspecific and traumatic synovitis, degenerative osteoarthritis or rheumatoid arthritis were 3.8 to 4.6g/dl. There was no significant difference between each from of arthritis. The pattern of protein fractionation of synovial fluid by paper electrophoresis was similar to that of serum protein. On disc electrophoresis, 20 fractions were identified in synovial fluid and the main fraction was albumin. Isoelectric focusing of the human serum with Ampholine carrier ampholyte in thin layer polyacrylamide gel revealed 27 protein fractions and five isoenzymes of amylase and two of them were the main fractions. In the synovial fluid 22 protein fractions and two isoenzymes of amylase, which had the same isoelectric points as the main fractions of serum, were noted. It is suggested that the isoamylases in the synovial fluid are a dialysate of plasma enzymes.
Arthritis, Rheumatoid/metabolism ; Human ; Osteoarthritis/metabolism ; Proteins/metabolism* ; Synovial Fluid/metabolism* ; Synovitis/metabolism

OBJECTIVETo determine the concentrations of interleukin-18 (IL-18), IL-6, IL-8, and prostaglandin E2 (PGE2) in the synovial fluid in patients with osteoarthritis (OA), and explore the role of IL-18 in the pathogenesis of OA.

METHODSThe synovial fluid was collected from 30 patients with knee OA, and the concentrations of IL-18 and the other cytokines were measured using enzyme-linked immunosorbent assay (ELISA). A linear regression was performed between IL-18 and the other cytokines.

RESULTSThe average IL-18 and PGE2 concentrations were 220-/+304 pg/ml and 89-/+104 pg/ml in the synovial fluid, respectively, and the two cytokines showed a positive correlation in the synovial fluid (r=0.628, P=0.001). The IL-18 concentration was also correlated to the concentrations of IL-6 (1200-/+1587 pg/ml, n=22; r=0.590, P=0.008) and IL-8 (5190-/+6024 pg/ml, n=9; r=0.776, P=0.014).

CONCLUSIONIL-18 can promote PGE2 production, which causes cartilage degradation in OA, thus therapies targeting this cytokine may prove an effective approach to early OA treatment.

Aged ; Dinoprostone ; biosynthesis ; Female ; Humans ; Interleukin-18 ; metabolism ; Male ; Middle Aged ; Osteoarthritis ; metabolism ; Synovial Fluid ; metabolism

OBJECTIVE: To detect the concentrations of interleukin-18 (IL-18) and prostaglandin E2(PGE2) in synovial fluid (SF), and to determine the role of IL-18 and PGE2 in osteoarthritis (OA) pathogenesis. METHODS: IL-18 and PGE2 were measured concurrently in synovial fluid samples from 54 patients with knee OA (OA group) and from 9 controls (control group). Quantitative determination of IL-18 was performed by enzyme-linked immunosorbent assay (ELISA). PGE2 was examined by inhibitory enzyme-linked immunosorbent assay. A linear regression between IL-18 and PGE2 was analysed. RESULTS: The concentrations of IL-18 and PGE2 in SF from the OA group were significantly higher than those from the control group (P<0.01). The average value of IL-18 in the control group was (28.768+/-13.575) x 10(-9)ng/L, and (72.303+/-40.130) x 10(-9)ng/L in the OA group (P<0.01); the average value of PGE2 in the control group was (24.697+/-7.814) x 10(-9)ng/L, and (42.302+/-23.818) x 10(-9)ng/L in the OA group (P<0.01). IL-18 was related with PGE2 in a linear curve fashion (the control group: r=0.76, P<0.001; the OA group: r=0.94, P<0.001). CONCLUSION IL-18 and PGE2 are significantly higher in the OA group than those in the control group, and they might take part in the cartilage degradation in OA pathogenesis. The increase of IL-18 might induce the increase of PGE2, and that might play an important role in OA pathogenesis.
Adult ; Aged ; Case-Control Studies ; Dinoprostone ; metabolism ; Female ; Humans ; Interleukin-18 ; metabolism ; Male ; Middle Aged ; Osteoarthritis ; metabolism ; Synovial Fluid ; metabolism

OBJECTIVETo investigate the clinical effects of moxibustion (Mox) in treating knee joint osteoarthritis, and to detect the change of hyaluronic acid (HA) level in serum and synovial fluid (SF) for evaluating its significance.

METHODSThirty OA patients were treated with Mox applied on inner and outer hsiyens and Ashi point for 10 min once a day, 5 times a week for 3 months and the therapeutic efficacy was evaluated. Serum and SF levels of HA were measured by radio-immunoassay before and after the 3-month treatment, and compared with those from 30 non-OA persons for normal control.

RESULTSAfter treatment, 19 patients (20 joints) out of the 30 patients were cured, the efficacy of Mox was evaluated as markedly effective in 8 patients on 10 joints, and as effective in 3 patients on 4 joints, the cure rate being 63.3%. Before treatment, HA level in serum (122.87 +/- 34.10 microg/L) was higher and in SF (0.98 +/- 0.17 g/L) was lower in OA patients than those in the normal controls (68.32 +/- 21.48 microg/L and 1.62 +/- 0.30 g/L, P<0.01), whereas after treatment, both the serum and SF levels of HA in patients changed toward normal range (70.29 +/- 27.30 microg/L and 1.58 +/- 0.26 g/L), showing insignificant difference as compared with those in the controls (P<0.01).

CONCLUSIONMox is an effective approach for treatment of OA, and the levels of HA in serum and SF can be taken as the quantitative indicators for evaluating the pathogenetic condition of OA patients.

Female ; Humans ; Hyaluronic Acid ; analysis ; blood ; Male ; Middle Aged ; Moxibustion ; Osteoarthritis, Knee ; blood ; metabolism ; therapy ; Synovial Fluid ; metabolism

Rheumatoid arthritis is a chronic inflammatory systemic disease of young or middle aged adults, characterized by destructive and proliferative changes in synovial membrane, periarticular structure, skeletal muscle and perineural sheaths. Eventually, joints are destroied, ankylosed and deformed. Moderate anemia is occured frequently in rheumatoid arthritis patients, and these patients show abnormalities of iron metabolism such as lower serum iron concentration and occurance and distribution of iron in the synovial membrane. It has been suggested that the mechanism of iron deposits in rheumatoid arthritis is continuous oozing of blood from vascular granulation tissue into the synovial cavity and this lead the patient to anemia. We collected samples from serum and knee joint fluid in 21 cases of rheumatoid arthritis for chemical estimation of the ferritin concentration by radioimmunoassay, and compared with that of osteoarthritis. The following results were obtained: 1. The mean hemoglobin concentration(11.4gm %) in rheumatoid arthritis was lower than osteoarthritis(13.4gm %). 2. The mean serum ferritin concentration(118. 4ng/ml) in rheumatoid arthritis was lower than osteoarthritis(135. 6ng/ml), on the contrary in synovial fluid rheumatoid arthritis(279.8ng/ml) showed higher than osteoarthritis(190. 4ng/ml). 3. The mean ratio of synovial fluid ferritin on serum ferritin was 2. 36 in rheumatoid arthritis, in contrast with l. 4 in osteoarthritis. There was significant correlation between the ferritin concentration in synovial fluid and serum. 4. Serial check of ferritin concentration in synovial fluid during treatment would be thought meaningful criteria for determination of progress.and effectiveness of treatment.
Adult ; Anemia ; Arthritis, Rheumatoid ; Ferritins ; Granulation Tissue ; Humans ; Iron ; Joints ; Knee Joint ; Metabolism ; Middle Aged ; Muscle, Skeletal ; Osteoarthritis ; Radioimmunoassay ; Synovial Fluid ; Synovial Membrane

OBJECTIVETo investigate the role of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in the pathogenesis and progression of Kashin-Beck disease (KBD) and primary osteoarthritis (OA).

METHODSThe synovium and synovial fluid of the knee joint were collected from 20 adult patients with KBD, 18 with OA and 19 with meniscus injury (controls). The expression of IL-1beta and TNF-alpha in the synovium were analyzed by immunohistochemistry staining, and the levels of IL-1beta and TNF-alpha in the synovial fluid were determined by enzyme-linked immunosorbent assay.

RESULTIL-1beta and TNF-alpha expressions in the synovium and their levels in the synovial fluid were significantly higher in patients with KBD and OA than in patients with meniscus injury (P<0.05), but comparable between KBD and OA groups (P>0.05).

CONCLUSIONIL-1beta and TNF-alpha may play an important role in the pathogenesis and progression of KBD and OA.

Adult ; Aged ; Cartilage, Articular ; metabolism ; Endemic Diseases ; Female ; Humans ; Interleukin-1beta ; metabolism ; Male ; Middle Aged ; Osteoarthritis ; etiology ; metabolism ; Synovial Fluid ; metabolism ; Synovial Membrane ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVE: To examine the expression of chemokine receptor CCR10 on monocytes/macrophages in the joints of patients with rheumatoid arthritis (RA), and to investigate the role of chemokine CCL28 and its receptor CCR10 in the migration of RA monocytes and its mechanism. METHODS: The expression of CCR10 in synovial tissues from 8 RA patients, 4 osteoarthritis (OA) patients, and 4 normal controls was analyzed by immunohistochemistry, and cell staining was scored on a 0-5 scales. Flow cytometry was used to measure the percentage of CCR10 positive cells in CD14⁺ monocytes from peripheral blood of 26 RA patients and 20 healthy controls, as well as from synovial fluid of 15 RA patients. The chemotactic migration of monocytes from RA patients and healthy controls in response to CCL28 was evaluated using an in vitro Transwell system. Western blotting was conducted to assess phosphorylation of the extracellular signal-regulated kinase (ERK) and protein kinase B (Akt) pathways in RA monocytes upon CCL28 treatment. RESULTS: CCR10 was predominantly expressed in RA synovial lining cells and sublining macrophages, endothelial cells, and lymphocytes. CCR10 expression was significantly increased on lining cells and sublining macrophages in RA synovial tissue compared with OA and normal synovial tissue (both P < 0.01). The patients with RA had markedly elevated expression of CCR10 on peripheral blood CD14⁺ monocytes compared with the healthy controls [(15.6±3.0)% vs. (7.7±3.8)%, P < 0.01]. CCR10 expression on synovial fluid monocytes from the RA patients was (32.0±15.0)%, which was significantly higher than that on RA peripheral blood monocytes (P < 0.01). In vitro, CCL28 caused significant migration of CD14⁺ monocytes from peripheral blood of the RA patients and the healthy controls at concentrations ranging from 10-100 μg/L (all P < 0.01). The presence of neutralizing antibody to CCR10 greatly suppressed CCL28-driven chemotaxis of RA monocytes (P < 0.01). Stimulation of RA monocytes with CCL28 induced a remarkable increase in phosphorylation of ERK and Akt (both P < 0.05). ERK inhibitor (U0126) and phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002) strongly reduced the migration of RA monocytes in response to CCL28 (both P < 0.01). CONCLUSION RA patients had increased CCR10 expression on peripheral blood, synovial fluid, and synovial tissue monocytes/macrophages. CCL28 ligation to CCR10 promoted RA monocyte migration through activation of the ERK and PI3K/Akt signaling pathways. The CCL28-CCR10 pathway could participate in monocyte recruitment into RA joints, thereby contributing to synovial inflammation and bone destruction.
Humans ; Monocytes/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Endothelial Cells/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Arthritis, Rheumatoid ; Synovial Membrane ; Chemokines, CC/metabolism* ; Synovial Fluid ; Osteoarthritis ; Receptors, CCR10/metabolism*

S100A8 and S100A9 are major leukocyte proteins, known as damage-associated molecular patterns, found at high concentrations in the synovial fluid of patients with rheumatoid arthritis (RA). A heterodimeric complex of S100A8/A9 is secreted by activated leukocytes and binds to Toll-like receptor 4, which mediates downstream signaling and promotes inflammation and autoimmunity. Serum and synovial fluid levels of S100A8/A9 are markedly higher in patients with RA than in patients with osteoarthritis or miscellaneous inflammatory arthritis. Serum levels of S100A8/A9 are significantly correlated with clinical and laboratory markers of inflammation, such as C-reactive protein, erythrocyte sedimentation rate, rheumatoid factor, and the Disease Activity Score for 28 joints. Significant correlations have also been found between S100A8/A9 and radiographic and clinical assessments of joint damage, such as hand radiographs and the Rheumatoid Arthritis Articular Damage score. In addition, among known inflammatory markers, S100A8/A9 has the strongest correlation with total sum scores of ultrasonography assessment. Furthermore, baseline levels of S100A8/A9 are independently associated with progression of joint destruction in longitudinal studies and are responsive to change during conventional and biologic treatments. These findings suggest S100A8/A9 to be a valuable diagnostic and prognostic biomarker for RA.
Arthritis, Rheumatoid/*blood/pathology/radiography ; Arthrography ; Biological Markers/blood ; Calgranulin A/*blood ; Calgranulin B/*blood ; Humans ; Joints/pathology ; Synovial Fluid/metabolism

Objective To investigate the expression of miRNA-140 in chondrocytes and synovial fluid of osteoarthritis (OA) patients, and explore the relationship between the miRNA-140 expression and OA severity.Methods This study enrolled 30 OA patients who underwent total knee arthroplasty for chondrocytes sampling and 30 OA patients who underwent intra-articular injection for synovial fluid sampling. All OA patients were grouped into mild [Kellgren and Lawrence (KL) grade 1-2], moderate (KL grade 3) and severe (KL grade 4), with 10 in each subgroups for each sampling purposes. 7 non-OA patients and 10 patients with knee injury were collected for cartilage and synovial fluid sampling respectively as control groups. Chondrocytes were isolated from the cartilage tissue and cultured in vitro. Quantitative real time PCR for miRNA-140 in chondrocytes and synovial fluid were performed, and the U6 snRNA was used as internal control. The expression difference of miRNA-140 among groups and correlation between the expression and the KL grade of OA were analysed using one-way ANOVA and Spearman test respectively. Results The expression of miRNA-140 in chondrocytes of knees in OA patients was reduced than that in normal knees, and the between-group difference was statistically significant (F=305.464, P<0.001). miRNA-140 could be detected in synovial fluid of both normal knees and OA knees, its relative expression level was reduced in synovial fluid of OA group compared with normal group, and the between-group difference was statistically significant as well (F=314.245, P<0.001). The relative expression level of miRNA-140 in both chondrocytes and synovial fluid were negatively correlated with the KL grade of OA(r=-0.969, P<0.001; r=-0.970, P<0.001). Conclusion miRNA-140 could be detected in chondrocytes and synovial fluid of OA patients, and its expression was negatively correlated with the severity of OA.
Adult ; Aged ; Cells, Cultured ; Chondrocytes ; metabolism ; Female ; Humans ; Knee Joint ; metabolism ; Male ; MicroRNAs ; analysis ; Middle Aged ; Osteoarthritis, Knee ; metabolism ; Real-Time Polymerase Chain Reaction ; Synovial Fluid ; metabolism

OBJECTIVETo investigate the role of natural killer-22 (NK-22) cells in the synovial fluid in the proliferation of fibroblast-like synoviocytes (FLS) in patients with rheumatoid arthritis (RA) and explore the possible signal pathway involved.

METHODSNK-22 cells in the SF of RA patients were sorted by flow cytometry. NK-22 cells were cultured for two weeks and the purity was detected by flow cytometry before stimulation with 20 ng/ml phorbol 12-myristate 13-acetate and 0.5 µmol/L ionomycin for 4 h. The level of interleukin-22 (IL-22) in the culture medium supernatant was then measured with ELISA. The proliferation of FLS in the presence of the culture supernatant of NK-22 cells was assessed with MTT assay at 24, 48 and 72 h, and the effect of IL-22 antibody on FLS proliferation was also observed. Real-time PCR and Western blotting were used to detect Stat3 mRNA and p-Stat3 protein levels, respectively, in the FLS exposed to rhIL-22 and AG490.

RESULTSNK-22 cells were successfully sorted by flow cytometry with a purity exceeding 90%. The levels of IL-22 in the supernatant of NKp44(+)NK cell culture averaged 1273.42∓254.48 pg/ml. The FLS proliferated rapidly 24, 48, and 72 h after the addition of culture supernatant of NK-22 cells (P<0.05). IL-22 antibody obviously inhibited the proliferation of FLS induced by NK-22 cell culture supernatant (P<0.05). Exposure of the FLS to rhIL-22 obviously increased cellular Stat3 expression levels, which were significantly lowered by the addition of AG490 (P<0.05).

CONCLUSIONNK-22 cells in the SF of RA patients can produce high concentrations of IL-22 to promote the proliferation of FLS through the STAT3 signal pathway.

Arthritis, Rheumatoid ; metabolism ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; cytology ; Humans ; Interleukins ; metabolism ; Killer Cells, Natural ; cytology ; metabolism ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Synovial Fluid ; cytology ; Synovial Membrane ; cytology