Alcohol Extract of Angelicae Pubescentis Radix Treats Gouty Arthritis Induced by MSU via NLRP3/ASC/Caspase-1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the efficacy and mechanism of the alcohol extract DH50 of Angelicae Pubescentis Radix in treating gouty arthritis induced by monosodium urate （MSU） crystals in vivo and in vitro. MethodFifty male SD rats were randomly assigned into five groups （n=10）： a normal group， a model group， a dexamethasone （DXMS， 0.07 mg·kg-1） group， and low- （DH50-D， 9 mg·kg-1） and high-dose （DH50-G， 18 mg·kg-1） DH50 groups. The rats in the normal group and model group were administrated with the same amount of pure water. On day 5， the gouty arthritis model was established by injecting MSU into the right ankle joint of rats. The toe volume and joint inflammation index were measured 4， 8， 24， and 48 h after modeling. The pathological changes of the synovial tissue were detected by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of tumor necrosis factor-alpha （TNF-α）， interleukin （IL）-1β， and IL-6 in the synovial tissue. Western blot was employed to measure the protein levels of NOD-like receptor protein 3 （NLRP3）， cysteine-aspartic protease-1 （Caspase-1）， apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain （ASC）， IL-1β， and cyclooxygenase-2 （COX-2） in the synovial tissue. Furthermore， the cell inflammation model was established with RAW264.7 cells stimulated with MSU （75 mg·L-1）. The cell experiments were carried out with 6 groups： a normal group， a model group， a positive drug （DXMS， 100 μmol·L-1） group， and low- （DH50-D， 25 mg·L-1）， medium- （DH50-Z， 50 mg·L-1）， and high-dose （DH50-G， 100 mg·L-1） DH50 groups. Methyl thiazolyl tetrazolium （MTT） assay was employed to determine the cell viability， ELISA to determine the content of TNF-α in the supernatant of cell culture， and Western blot to determine the protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2. ResultCompared with the normal group， the rat model group showed increased toe swelling degree and joint inflammatory index （P<0.01）， serious infiltration of the synovium， elevated levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and up-regulated protein levels of NLRP3， Caspase-1， ASC， IL-1β， and COX-2 （P<0.05， P<0.01）. Compared with the rat model group， low- and high-dose DH50 mitigated the toe swelling degree， decreased the joint inflammatory index， alleviated the inflammatory infiltration， lowered the levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and down-regulated the expression of related proteins （P<0.05， P<0.01）. Compared with the normal group， the cell model group showed elevated level of TNF-α in the supernatant （P<0.01） and up-regulated protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2 （P<0.05）. Compared with the model group， low， medium， and high doses of DH50 lowered the level of TNF-α in the supernatant of cell culture in a dose-dependent manner and down-regulated the expression of related proteins （P<0.05， P<0.01）. ConclusionDH50 can mitigate gouty arthritis both in vitro and in vivo by inhibiting the activation of NLRP3 inflammasomes and the production of inflammatory cytokines.

Nasal gouty tophus

@#A 48-year-old, non-hypertensive, non diabetic man with uncontrolled gouty arthritis presented with a four-day swollen nasal mass. He was assessed to have a nasal abscess at the emergency room and was admitted for urgent management. Paranasal computed tomography (CT) scans showed a heterogeneously enhancing focus with areas of hypodensities in the nasal apex and dorsum extending into the right ala measuring 1.5 x 2.8 x 3.4 cm. with associated erosion of the cartilaginous part of the anterior nasal septum, soft tissue swelling and skin thickening in the nasal dorsum, nasal tip and right zygomatic region that was suspected to relate to an aggressive etiology. Tissue correlation was therefore recommended, and he underwent endoscopic-guided incision and drainage with biopsy and debridement of the nasal mass. The specimen submitted consisted of red to white, irregular, soft tissue fragments with an aggregate measurement of 1.5 x 1.5 x 0.5 cm. Microsections showed deposits of amorphous white to pink material with surrounding fibrosis and acute and chronic inflammatory cell infiltrates and foreign body giant cells. (Figures 1 and 2) Also seen in the background were fragments of sclerotic bone and bacterial colonies. These findings were consistent with gouty tophus with acute and chronic inflammation and bacterial colonization. The culture and sensitivity test of the nasal discharge showed growth of Enterobacter aerogenes (currently named Klebsiella aerogenes) which was identified by an automated mass spectrometry microbial identification system (VITEK® MS). Work-up also included uric acid levels which were within the reference interval at that time (6.57 mg/dL).

Pathogenesis of Gouty Arthritis and Intervention with Chinese Medicines： A Review / 中国实验方剂学杂志

Gouty arthritis （GA） is the metabolic rheumatism caused by purine metabolism disorder， which can be acute or chronic. The main manifestations of GA include recurrent redness， swelling， heat pain， and dysfunction of the affected joints. According to the theory of modern medicine， GA is closely associated with the increase in uric acid， the participation of inflammatory cytokines， the weakening of antioxidant response， apoptosis， and the imbalance of intestinal flora and bone metabolism， whereas the specific pathogenesis remains unclear. GA is characterized by easy diagnosis， difficult treatment， and high recurrence rate， which seriously affects the life quality of patients. Colchicine， corticosteroids， non-steroidal anti-inflammatory drugs， and selective cyclooxygenase-2 inhibitors are the commonly used western medicines for this disease， which demonstrate remarkable short-term therapeutic effect. However， long-term use of these medicines will bring serious adverse reactions. Chinese medicines， with high safety and causing few adverse reactions， have a variety of active components which can act on multiple pathways and targets to exert synergistic effects， thus attracting wide attention. This paper systematically reviews the literature reporting the Chinese medicines in improving antioxidant response， reducing chondrocyte apoptosis， and regulating intestinal flora and bone metabolism， aiming to further clarify the pathogenesis of GA and provide a scientific basis for the clinical application of Chinese medicines in the prevention and treatment of GA.

What is the diagnostic value of dual-energy computed tomography in patients with clinical diagnosis of gout?

Abstract Objectives: To investigate the frequency of monosodium urate (MSU) crystal deposits on dual-energy computed tomography (DECT) in patients with clinical diagnosis of gout and the factors associated MSU crystal positivity. Methods: This study was conducted in patients with clinical diagnosis of gout who underwent DECT. Clinical features were compared between patients with positive and those with negative DECT results. A logistic regression analysis was performed to determine the factors associated with MSU crystal positivity on DECT. Results: A total of 148 patients with clinical diagnosis of gout were included, and MSU crystal deposition on DECT was observed in 64 patients (43.3%). The patients with positive DECT results were more likely to have renal insufficiency, longer disease duration, and higher serum urate level than those with negative. In the multivariable analysis, first gout attack (odds ratio 0.462; 95% confidence interval 0.229-0.931, p = 0.031) was associated with a less likely MSU crystal deposit-positive DECT result. In the subgroup analysis of patients with first attack, serum urate level > 8 mg/dL was associated with DECT positivity. Conclusion: Of the patients with clinical diagnosis of gout, those with renal insufficiency, longer disease duration, and high serum urate level were more likely to be positive of gout on DECT. First gout attack was associated with less likely to be positive for MSU crystal on DECT. Thus, performing DECT scan in the selected patients who had characteristics that highly probability of DECT positivity could increase positive predictive value.

Protective effects of Zingiber zerumbet rhizome extract on monosodium urate crystal-induced gout rat model

Gout is the most common inflammatory arthritis worldwide. Untreated gout causes severe pain and affects the qualityof life. Zingiber zerumbet is a well-known traditional plant with anti-inflammatory and antioxidant properties. Thisstudy was conducted to evaluate the effects of Z. zerumbet extract in gout-induced rats. In this study, 30 male Sprague–Dawley rats were divided into five groups (six rats in each group): i) normal control group received normal salineand tween 80, ii) model group injected with monosodium urate (MSU) crystal solution, iii) positive control groupreceived 10 mg/kg celecoxib drug, iv) Z. zerumbet extract at 200 mg/kg, and v) Z. zerumbet extract at 400 mg/kg. Theextract was orally administered for 14 days. Rats of all groups (except the normal control group) were injected withMSU crystals (50 μl) into the ankle joint on day 11. The ankle joint was measured before injection 10, 24, 48, and 72hours after injection. On day 14, all rats were sacrificed. Blood and tissue samples were obtained for the analysis ofinflammatory and oxidative stress biomarkers. In this study, the ankle joint diameter rate was significantly reducedby Z. zerumbet at 200 and 400 mg/kg (p < 0.05) compared to the model group. The extract prevented inflammationsignificantly (p < 0.05) compared to the model group in white blood cell count, C-reactive protein, and erythrocytesedimentation rate tests. The two doses of Z. zerumbet also significantly (p < 0.05) promoted the activity of superoxidedismutase and attenuated 8-isoprostane. Thus, this outcome shows that the administration of Z. zerumbet rhizomeethyl acetate extract may be useful and easy to protect against gouty arthritis and the process is probably mediatedthrough its anti-inflammatory and antioxidant properties.

Analysis on Urine Metabolomics of Dioscoreae Nipponicae Rhizoma Extract Against Acute Gouty Arthritis / 中国实验方剂学杂志

Objective::To explore the effect of Dioscoreae Nipponicae Rhizoma extract (DNRe) on rats with acute gouty arthritis (AGA) based on urine metabolomics and to search for the related potential biomarkers and metabolic pathways. Method::Rat model of AGA induced by monosodium urate (MSU) was selected, 40 rats were randomly divided into the blank group (k), the DNRe group (g), the model group (m), and the DNRe treatment group (gm), with 10 rats in each group. The drug-administered group was administered with DNRe at a dose of 0.48 g·kg-1 once a day for 5 days. The urine was gathered after the last administration, and analyzed with UPLC-Q-TOF/MS coupled with pattern recognition techniques, electrospray ionization (ESI) under positive and negative ion scanning mode was adopted, data collection range was m/z 100-1 500 with full scanning mode. Result::A total of 12 common potential biomarkers were identified as sarcosine, dimethylglycine, deoxycytidine, uric acid, 5-hydroxytryptamine (5-HT), L-cystathionine, 4-pyridoxic acid, deoxyuridine, melatonin, 5-methoxytryptamine, fumaric acid and cytidine. Compared with the blank group, the 12 potential biomarkers in the DNRe group were significantly down-regulated. Compare with the model group, 10 metabolites were up-regulated and 2 metabolites were down-regulated in the 12 potential biomarkers of the DNRe treatment group, the abnormal expression of 10 markers including sarcosine, uric acid, L-cystathionine, 4-pyridoxic acid, deoxyuridine, 5-methoxytryptamine, cytidine, dimethylglycine, melatonin, fumaric acid could be modulated by DNRe. The strongest metabolic pathways associated with AGA were cysteine and methionine metabolism, and tryptophan metabolism. Conclusion::The effect of DNRe on AGA may be related to the promotion of conversion level from cystathionine to cysteine in the cysteine and methionine metabolism, and the up-regulating melatonin level in tryptophan metabolism.

Effect of Total Saponin of Dioscoreae Collettii Rhizoma on Toll-like Receptor/Nuclear Factor-κB (TLR/NF-κB) Signaling Pathway Induced by Monosodium Urate in THP-1 Cells / 中国实验方剂学杂志

Objective::To investigate the effect of total saponin of Dioscoreae Collettii Rhizoma (TSD) on Toll-like receptor/nuclear factor-κB (TLR/NF-κB) signaling pathway induced by monosodium urate in THP-1 cells, in order to explore the possible mechanism of anti-gout arthritis. Method::Phorbol 12-myristate 13-acetate (PMA)-induced THP-1 cells were differentiated into macrophages, divided into normal group, model group, low, medium and high-concentration TSD groups (1, 3, 10 mg·L-1) and colchicine group (0.2 mg·L-1). Except the normal group, the other groups were stimulated with 400 mg·L-1 monosodium urate to replicate an inflammation model in vitro. Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay, the levels of inflammatory factors tumor necrosis factor-α(TNF-α ) and interleukin-1β(IL-1β) were detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and NF-κB were detected by Western blot. The mRNA levels of TLR4, NF-κB and Pro-IL-1β were measured by real-time fluorescence quantitative PCR (Real-time PCR), and the nuclear shift of NF-κB p65 was detected by immunofluorescence. Result::0~32 mg·L-1 TSD has no effect on cell viability. Compared with the normal group, the secretion levels of inflammatory factors TNF-α and IL-1β in the model group were significantly increased (P<0.01), and the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were increased (P<0.01). Compared with the model group, 1-30 mg·L-1 TSD significantly down-regulated the secretion of inflammatory factors TNF-α and IL-1β (P<0.01), the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were decreased (P<0.05, P<0.01), and the NF-κB p65 partially trans-located to the cytosol and the superposition in the nucleus were decreased, inhibiting the nuclear translocation of NF-κB p65. Conclusion::TSD may exert an anti-inflammatory effect by down-regulating the expressions of TLR4, NF-κB and Pro-IL-1β mRNA and reducing the secretion of inflammatory factors TNF-α and IL-1β.

Influence of Bacopa monnieri in suppressing the arthritis induced by MSU crystal in Wistar albino female rats: Through biochemical and histopathological approach

Gouty arthritis is caused due to the accumulation of uric acid crystals in joints which is the by-product of purinemetabolism in our body. The elevated level of uric acid in the blood is known as hyperuricemia that may resultsin deposition and inflammation of joints. This research experiment has examined the protective effect of Bacopamonnieri, an herb against the monosodium urate crystal-induced gouty arthritis in female Wistar albino rats. The ratswere divided into four groups with six rats in each group. Group-I was normal control rats, group-II rats were inducedwith monosodium urate crystal, group-III was administrated with B. monnieri in monosodium urate crystal-inducedrats, and group-IV was administrated with indomethacin in monosodium urate crystal-induced rats. The rats wereexamined for liver enzyme markers, antioxidant assays, and paw histopathology. The results of B. monnieri werecompared with that of standard drug indomethacin. The symptoms of arthritis, such as the elevation of paw volume,increased liver enzyme markers, decreased antioxidant enzymes, and histopathological changes, were found to bereversed to the normal level by the treatment with B. monnieri which is due to its anti-inflammatory properties. Asconclusion, B. monnieri has shown its anti-arthritic properties against gouty arthritis.

Expression and significance of miR-21 in primary gout patients / 中华风湿病学杂志

Objective To explore the expression and significance of miR-21 in patients with primary gout. Methods The patients were divided into 4 groups: 35 acute gout patients (AG), 50 intermittent gout patients (IG), 25 chronic gout patients (CG) and 39 healthy patients. Their peripheral blood were collected and laboratory indexes were recorded. The expression of miR-21 and Nod-like receptor pyrin domain-containing protein 3 (NLRP3) mRNA in the peripheral blood mononuclear cells (PBMCs) was detected by real-time quantitative polymerase chain reaction (RT-qPCR). The blood and clinical data of another 5 healthy volunteers were collected, their peripheral blood was stimulated with 100 μg/ml monosodium urate (MSU) for 1 hour, pho-sphate buffer (PBS) was used as controls, then the expression of microRNA (miR)-21, NLRP3, interleukin (IL)-1β mRNA was detected by RT-qPCR. Rank sum test and spearman correlation analysis were used for data analysis. Results In primary gout patients, the expression of miR-21 in AG [12 ×10-4 (8.0 ×10-4)], IG [9.4 ×10-4 (6.9 ×10-4)], CG [7.3 ×10-4 (5.6 ×10-4)] was significantly higher than that in healthy control group [1.0×10-4(2.0×10-4)] (Z=9.83, P=0.02], while the expression of NLRP3 in AG[0.0444(0.0233)], IG[0.0581(0.0326)], CG[0.0314(0.0198)] was significantly lower than that in healthy control group [0.0886(0.0359)] (Z=13.82, P<0.01). In the primary gout of IG group, the expression of miR-21 was positively correlated with NLRP3 mRNA (r=0.449, P=0.016). After stimulated by 100 μg/ml MSU, the expression of miR-21 of the stimulated group [8.78×10-4(14×10-4)] was higher than that in the control group [6.25×10-4(6×10-4)](Z=-2.203, P<0.05), and the expression of IL-1βin stimulated group [3.06(2.00)] was higher than that in the control group [2.64 (1.22] (Z=-2.203, P<0.05). The level of miR-21 in patients with primary gout was positively correlated with the level of uric acid (UA), glutamic oxaloacetic transaminase (AST) and glutamic pyruvic transaminase (ALT) (r=0.473, 0.639, 0.487, P<0.05). Conclusion The increase of miR-21 in patients with primary gout may be involved in the inflammatory reaction of gout.

Relationship between Urate Crystal Deposits Detected by Dual-energy Computed Tomography and Bone Erosions in Symptomatic Gout Patients without Clinically Apparent Tophi

OBJECTIVE: Dual-energy computed tomography (DECT) allows sensitive detection of monosodium urate (MSU) crystal deposits in gout. However, the role of MSU deposits on DECT during the disease process of gout is not clear. The aim of our study was to evaluate the relationship between joint damage and MSU deposits detected by DECT in symptomatic non-tophaceous gout. METHODS: DECT scans of 51 gout patients without clinically apparent tophi were assessed. Individual ankle and foot joints and Achilles tendon insertion sites were evaluated for the presence of MSU deposits and bone erosions. The total volume of MSU crystal on DECT was quantified using an automated software program. Clinical and laboratory data at the time of the DECT evaluation were obtained from medical record. RESULTS: MSU deposits were detected in 92.2% of the patients evaluated. Median number and total volume of MSU deposit per patient was 5.0 and 0.6 cm3, respectively. Bone erosion was found in 54.9% of patients. MSU deposits in the first (1st) metatarsophalangeal (MTP) joints were significantly associated with presence of bone erosions (odds ratio [OR] 3.77, 95% confidence interval [CI] 1.06~13.38, p=0.040). Older age and frequent gout attack were associated with development of bone erosion in patients with MSU deposits (OR 1.12 and 2.57, 95% CI 1.04~1.22 and 1.02~6.50, p-value 0.004 and 0.047, respectively). CONCLUSION: MSU deposits and erosions were frequently detected by DECT in symptomatic non-tophaceous gout patients, and MSU deposits in 1st MTP joints were associated with presence of bone erosions especially in patients with older age and frequent gout attack.

Performance of the 2015 American College of Rheumatology/European League against Rheumatism Classification Criteria for Gout in Korean Patients with Acute Arthritis

BACKGROUND: We aimed to assess the performance of the 2015 American College of Rheumatology (ACR)/European League Against Rheumatism (EULAR) classification criteria for gout in Korean patients with acute arthritis and to compare the performance of the ACR/EULAR criteria to that of other sets of criteria for gout classification. METHODS: Patients with acute arthritis who underwent diagnostic arthrocentesis at one of the four participating rheumatology clinics were consecutively enrolled between February and December 2017. Crystal-proven gout was diagnosed upon confirming the presence of monosodium urate (MSU) crystals in patients with a clinical impression of gout as judged by the rheumatologist. The performance of the ACR/EULAR and other gout classification criteria, including the Rome, New York, American Rheumatism Association (ARA), Mexico, and Netherlands criteria, was analyzed regardless of the presence/absence of MSU crystals. RESULTS: The study enrolled 118 gout patients (all crystal-proven) and 95 non-gout patients. According to the area under the curve, the diagnostic performance was the highest for the ACR/EULAR classification criteria (sensitivity, 80.5%; specificity, 95.8%; area under the curve, 0.966), followed by the Netherlands, Rome, ARA, New York, and Mexico criteria. All six sets of criteria demonstrated lower sensitivity in patients exhibiting the first episode of acute arthritis. CONCLUSION: In Korean patients with acute arthritis, the ACR/EULAR classification criteria outperformed other sets of gout classification criteria even in the absence of information regarding the presence of MSU crystals. However, to enhance diagnostic sensitivity, synovial fluid analysis should be considered in patients with the first episode of acute arthritis.

Expression and clinical significance of long noncoding RNA-AK001903 in patients with gouty arthritis / 中华风湿病学杂志

Objective To investigate the role of long noncoding RNA-AK001903 in the pathogenesis of primary gout arthritis (GA).Methods The subjects were divided into four groups:30 acute gout patients (AGA),24 non-acute gout patients (NAGA),24 healthy controls and 24 hyperuricemia (HUA).Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to examine the expression of AK001903 in peripheral blood mononuclear cells (PBMCs) from four groups.100 μg/ml monosodium (MSU) was used to stimulate the peripheral blood of NAGA and healthy control patients.Then the expression of AK001903 was detected by RT-qPCR.Kruskal-Wallis test,Mann-Whitney test,Spearman correlations were used for statistical analysis.Results The expression level of AK001903 in the AGA group (0.079±0.022) and the NAGA group (0.071±0.021) were higher than the healthy control group (0.014±0.004).There was no significant difference between the NAGA group and the NAGA group (Z=-0.655,P＞0.05).Those of the GA group (0.078±0.018) and the HUA group(0.047±0.016) was higher than the healthy control group (0.014±0.004) (Z=-2.887,Z=-4.157;P＜0.05).Compared with the control group,the expression of AK001903 in NAGA and the healthy control group which were stimulated by MSU was significantly increased.The Spearman correlation analysis found that the AK001903 expression levels in the GA groups were correlated with TG (r=0.938,P＜0.05),VLDL (r=0.873,P＜0.05),GLU9 (r=0.671,P＜0.05) and were negatively correlated with apoA1 (r=-0.661,P＜0.05).Conclusion Altered expression of AK001903 may be involved in the process of imbalance between lipid metabolism and hyperuricemia,and takes part in the pathogenesis of GA.

Altered expression of miR-223 might be involved in regulation of inflammation in primary gout patients / 中华风湿病学杂志

Objective To investigate the role of miR-223 in the pathogenesis of acute gouty inflammation.Methods The subjects were divided into 3 groups:65 acute gout patients (AG),50 inter-critical gout patients (IG),and 45 healthy controls (HC).The peripheral blood mononuclear cells (PBMCs) and the clinical laboratory parameters were all collected.The expression of miR-223 in the PBMCs was detected using realtime fluorescent quantitative polymerase chain reaction (RT-qPCR) (TaqMan probe).The PBMCs of 5 healthy people were stimulated with monosodium urate (MSU) (100 μg/ml) for 12 h,and then,miR-223,NLRP3 mRNA and IL-1β production were all measured using RT-qPCR and ELISA respectively.All data were analyzed by SPSS 17.0 statistical software,Wilcoxon rank sum test,t test and Spearman's correlations analysis were used for statistical analysis.Results ① The expression of miR-223 in AG and IG groups was both significantly decreased than that in the HC group (8±17 vs 26±76,P＜0.05;9±17 vs 26±76,P＜0.05;respectively),AG group was significantly decreased than that in the IG group [8(17) vs 9(17),Z=11.387,P＜0.01].② After stimulated with MSU in healthy controls,IL-1β production and NLRP3 mRNA were both significantly increased [(86±5) pg/ml vs (13±6) pg/ml,t=21.042,P＜0.01;5.2±0.4 vs 1.2±0.4,t=14.640,P＜0.01;respectively],while the expression of miR-223 was significantly decreased (0.34±0.20 vs 1.05±0.24,t=-5.164,P＜0.01).Conclusion The data suggests that miR-223 might be involved in the patho-genesis of spontaneous regulation,but further study is needed to discover the exact mechanism.

Establishment of a rat model of acute gouty arthritis and observation of the model maintenance time / 中国实验动物学报

Objective To establish a model of acute gouty arthritis( AGA) in rats and observe its maintenance time. Methods The AGA model of rats was established by injecting monosodium urate ( MSU) at the concentration of 25 mg/mL into the ankle joint cavity. The rats were observed for 8 d at different time points. Skin temperature, degree of joint swelling, gait, inflammatory cells in synovial fluid, histopathological changes of synovial tissue and other indicators were observed to determine whether the modeling and maintenance time were successful. Results At 3 h after modeling, differ-ences in the swelling of ankle joint, increase of skin temperature, abnormal gait, the number of inflammatory cells in syno-vial fluid, synovial hyperplasia, capillary congestion, and disarrangement of synovial cells in the rats were observed in the saline group and the model group (P <0. 01). At 4 hours after modeling, the above mentioned inflammatory changes in the saline group were significantly reduced, compared with that at 3 h, showing a significant difference (P<0. 01), while the inflammatory changes of the model group were increased significantly compared with that at 3 hours ( P<0. 01 ) , and showed significant difference compared with the saline group (P<0. 01). At 24 h after modeling, the indexes in the rats of saline group returned to normal, but the inflammation of the model group was increased. At 48-72 h after modeling, the local inflammation such as ankle swelling, skin temperature, and abnormal gait of the rats in the model group reached a peak. The inflammation of the ankle joint in the model group was gradually reduced from 96 to 168 h after the model was established, but there were still significant differences in the indexes compared with the blank group (P<0. 01). At 192 h after modeling, the joint swelling, skin temperature and abnormal gait of the rats in the model group returned to normal, however, there were significant differences in the number of inflammatory cells and the pathological changes of synovial membrane compared with the blank group ( P<0. 01 ) . Conclusions A rat model of AGA can be successfully prepared and identified at 4 h after modeling by injection of MSU crystal suspension into the ankle joint cavity. This rat model of AGA can be maintained at least 168 hours after modeling.

Establishment of a rat model of acute gouty arthritis and observation of the model maintenance time / 中国实验动物学报

Objective To establish a model of acute gouty arthritis( AGA) in rats and observe its maintenance time. Methods The AGA model of rats was established by injecting monosodium urate ( MSU) at the concentration of 25 mg/mL into the ankle joint cavity. The rats were observed for 8 d at different time points. Skin temperature, degree of joint swelling, gait, inflammatory cells in synovial fluid, histopathological changes of synovial tissue and other indicators were observed to determine whether the modeling and maintenance time were successful. Results At 3 h after modeling, differ-ences in the swelling of ankle joint, increase of skin temperature, abnormal gait, the number of inflammatory cells in syno-vial fluid, synovial hyperplasia, capillary congestion, and disarrangement of synovial cells in the rats were observed in the saline group and the model group (P <0. 01). At 4 hours after modeling, the above mentioned inflammatory changes in the saline group were significantly reduced, compared with that at 3 h, showing a significant difference (P<0. 01), while the inflammatory changes of the model group were increased significantly compared with that at 3 hours ( P<0. 01 ) , and showed significant difference compared with the saline group (P<0. 01). At 24 h after modeling, the indexes in the rats of saline group returned to normal, but the inflammation of the model group was increased. At 48-72 h after modeling, the local inflammation such as ankle swelling, skin temperature, and abnormal gait of the rats in the model group reached a peak. The inflammation of the ankle joint in the model group was gradually reduced from 96 to 168 h after the model was established, but there were still significant differences in the indexes compared with the blank group (P<0. 01). At 192 h after modeling, the joint swelling, skin temperature and abnormal gait of the rats in the model group returned to normal, however, there were significant differences in the number of inflammatory cells and the pathological changes of synovial membrane compared with the blank group ( P<0. 01 ) . Conclusions A rat model of AGA can be successfully prepared and identified at 4 h after modeling by injection of MSU crystal suspension into the ankle joint cavity. This rat model of AGA can be maintained at least 168 hours after modeling.

Liver metabonomics of acute gouty arthritis treated by Diosocorea nipponica / 中国中药杂志

To explore the prevention and protection effect of Diosocorea nipponica (DNM)) on acute gouty arthritis (AGA) rats based on liver metabonomics, and find potential biomarkers and related pathways. AGA model rats were induced by monosodium urate crystal suspension. UPLC-TOF-MS coupled with pattern recognition technique was employed to find out the potential biomarkers and related metabolic pathways. Eleven common potential biomarkers were identified. Among the potential intervention targets in normal rats given by DNM, 4 biomarkers were up-regulated, and the other 4 targets were down regulated. Among the potential intervention targets in AGA rats given by DNM, 5 metabolites were up-regulated by MSU and 5 metabolites were down regulated. The abnormal expression levels of adenosine monophosphate, 5-methyltetrahydrofolic acid, oxidized glutathione, hypoxanthine, docosahexaenoic acid, glutathione, uridine diphosphate glucose and inosine could be corrected by DNM extract. Three pathways were founded with greatest correlation, including purine metabolism, starch and sucrose metabolism and glutathione metabolism. Therefore, it could be inferred that D. nipponica has the effect for anti-acute gouty arthritis by intervening endogenous metabolites from the liver under physiological condition and acute gouty arthritis condition.

Anti-inflammatory effects of traditional mixed extract of medicinal herbs (MEMH) on monosodium urate crystal-induced gouty arthritis / 中国天然药物

Korean oriental medicine prescription is widely used for the treatment of gouty diseases. In the present study, we investigated anti-inflammatory effects of modified Korean herbal formulation, mixed extract of medicinal herbs (MEMH), and its modulatory effects on inflammatory mediators associated with gouty arthritis. Both in vitro and in vivo studies were carried out to assess the anti-inflammatory efficacy of MEMH on monosodium urate (MSU) crystals-induced gouty inflammation. MSU crystals stimulated human chondrosarcoma cell line, SW1353, and human primary chondrocytes were treated with MEMH in vitro. The expression levels of pro-inflammatory mediators and metalloproteases were analyzed. The effect of MEMH on NFκB signaling pathway in SW1353 cells was examined. Effect of MEMH on the mRNA expression level of pro-inflammatory mediators and chemotactic factor from human monocytic cell line, THP-1, was also analyzed. The probable role of MEMH in the differentiation process of osteoblast like cells, SaOS-2, after MSU treatment was also observed. To investigate the effects of MEMH in vivo, MSU crystals-induced ankle arthritic model was established. Histopathological changes in affected joints and plasma levels of pro-inflammatory mediators (IL-1β and TNFα) were recorded. MEMH inhibited NFκB signaling pathway and COX-2 protein expression in chondrocytes. MSU-induced mRNA expressions of pro-inflammatory mediators and chemotactic cytokines were suppressed by MEMH. In MSU crystals-induced ankle arthritic mouse model, administration of MEMH relieved inflammatory symptoms and decreased the plasma levels of IL-1β and TNFα. The results indicated that MEMH can effectively inhibit the expression of inflammatory mediators in gouty arthritis, demonstrating its potential for treating gouty arthritis.

Significance of mtDNA expression in acute peritonitis induced by monosodium urate in mice / 中华风湿病学杂志

Objective To establish acute peritonitis induced by monosodium urate (MSU) of in mice and observe the significance of mitochondrial deoxyribonucleic acid (mtDNA) expression in the inflammatory processes.Methods The mouse models of acute peritonitis were made by intraperitoneal injection of MSU.Sixty-four male C57BL16 mice were randomly divided into the MSU group which were treated with 0.2 ml of 15 mg/ml MSU solution by i.p.injection and the control group which were treated with 0.2 ml of PBS.Respectively four mice from MSU group and four mice from control group were killed 2 hours, 4 hours, 6 hours, 8 hours 12 hour, 16 hours, 20 hours and 24 hours later and whole blood, peritoneal lavage and peritoneum were collected respectively.Four the mice from the MSU group and four mice from the control group were killed and whole blood, peritoneal lavage and peritoneum were collected.Immunoflourescence study of peritoneum tissues was performed.The levels of interleukin (IL)-1β, IL-18 in plasma and peritoneal lavage were examined by enzyme linked immunosorbent assay (ELISA).DNA was extracted from blood and peritoneal lavage, and mtDNA level was detected by using real-time polymerase chain reaction (PCR).The data was analyszied by multivariate analysis of variance.Results As compared with those killed at other time points from the MSU groups and the control group, the levels of IL-1β [(27.0±2.0) pg/ml vs (26.8±2.1) pg/ml], IL-18 [(673±454) pg/ml vs(752±495) pg/ml] in plasma and peritoneal lavage were increased progressively in those which were killed after i.p.injection of 2 hours and 4 hours from in the MSU group (F=22.778, P＜0.05;F=6.660, P＜0.05).The mtDNA in plasma and peritoneal lavage of the mice began to be expressed 4 hours after i.p.injection 4 hours from in the MSU group.The peak level was detected in those i.p.injected MSU 6 hours later [(9.85±4.59)×106 copies, (7.81±3.43)×106 copies].Then 8 hours later the mtDNA began to slowly decreased.At these three time points, the mtDNA were all increased progressively than those at the other time points of the MSN group or at all time points of the control group (F=6.719, P＜0.05;F=11.181, P＜0.05).By immunoflourescence study, there were neutrophil extracellular traps (NETs) were formed 12 hours later in the MSU group and aggregated NETs were found 24 hours later.Conclusion In the inflammatory processes of acute peritonitis induced by MSU of in mice, with the expression of mtDNA increasing, the inflammation is relieved, and aggregated NETs are formed in the end.Expression of mtDNA may be one for the protective factors of the inflammation induced by MSU.

Distal Interphalangial Joint Gouty Arthritis in a Patient with Nodal Osteoarthritis: A Case Report and Review of Literature.

Case: Herein 73 year- old female patient with distal interphalangial (DIP) joint gouty arthritis accompanying nodal OA was presented. There was significant redness and swelling in the joint, joint was warm and tender on palpation and range of motion was very painful. Conclusion: The presentation of acute or subacute arthritis in interphalangial joints of a woman with preexisting nodal OA may obscure the correct diagnosis and coexisting gouty arthritis may be overlooked.

Cytology of Synovial Fluid in Gouty Arthritis: Two Cases Report

Synovial fluid (SF) aspiration cytology is a useful diagnostic tool. For patients with gouty arthritis, the diagnosis is confirmed by the presence of monosodium uric acid (MSU) crystals in the SF, and these crystals are long, pointed ended and needle-shaped and they show strongly negative birefringence. Sometimes, it is difficult to diagnosis between gouty arthritis and other type of inflammatory arthritis. We experienced two unusual cases of gouty arthritis that we performed SF analysis for. The first patient was a 35 year old male who presented with relatively typical clinical symptoms with hyperuricemia, but the SF showed acute inflammatory cells without crystals on light microscopy. Only a few suspected crystals of MSU were identified on polarizing microscopy. The second patient was a 45 year old male who presented with atypical symptoms and pain and swelling of the left ankle and knee joint for 3 weeks. The uric acid level in the serum and urine was increased, but not over the normal limit. However, on light and polarizing microscopy, there were numerous MSU crystals in the SF. Conclusively, in some cases of gouty arthritis, the crystals are not identified on light microscopy or the uric acid level is not dramatically increased. So, the polarizing microscopy, the clinical information and the laboratory findings are all included in the work-up when evaluating the SF cytology of arthritis patients.