Objective:To investigate the effect of stilamin on mitochondrial injury of acute pancreatitis (AP)-related acinar cells and the possible mechanism.Methods:24 C57BL/6 mice were randomly divided into control group, AP group and AP+ stilamin treatment group (Stilamin group). AP model was prepared by intraperitoneal injection of l-arginine in AP group, 0.4 mg/kg stilamin was intraperitoneally injected at 2 h after modeling in stilamin group, and control group received intraperitoneal injection with saline in the same volume. Histopathological examination of pancreatic tissue was performed routinely. Serum levels of IL-6, IL-10 and TNF- α were detected by ELISA, serum levels of amylase and lipase, and serum levels of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) reflecting oxidative stress were detected by biochemistry method, respectively. Mitotracker red fluorescent labeling was used to detect the number of mitochondria in pancreatic acinar cells, and western blot was used to detect the ND-3 protein expression reflecting the number of mitochondria. The expression levels of mitochondrial fusion protein (Mfn-2) and mitochondrial transcription factor A (TFAM) reflecting mitochondrial function were determined by immunohistochemical staining.Results:Compared with AP group, the pancreatic pathology scores of mice in Stilamin group were significantly decreased [(2.07±0.50) vs (3.93±0.64)], serum amylase and lipase levels were significantly decreased [(1 493±172)U/L vs (1 832±86)U/L, (225.4±83.2)U/L vs (671.0±164.5)U/L]; serum IL-6 and TNF-α levels were greatly decreased, while IL-10 levels were obviously increased [(99.09±39.65)ng/L vs (358.60±139.22)ng/L, (22.75±11.24)ng/L vs (40.83±1.62)ng/L, (15.12±5.03)ng/L vs (9.92±8.73)ng/L]. Mitotracker staining density and expression levels of ND-3, Mfn-2 and Tfam were increased [(71.67±17.62) vs (40.00±10.15), (0.45±0.16) vs (0.11±0.05), 78% vs 54%, 86% vs 47%], and MDA, SOD, and GSH levels were increased [(5.00±1.73)nmol/mg vs (7.33±2.08)nmol/mg protein, (17.33±3.21)U/mg vs (8.67±2.07)] U/mg protein. The ratio of (131.33±20.55)U/mg to (77.33±29.69)U/mg protein was statistically significant (all P<0.05). Conclusions:By protecting the mitochondria number and function of damaged pancreatic acinar cells and reducing the level of oxidative stress, stilamin could alleviate the level of pancreatic tissue damage and inflammatory response in mice with acute pancreatitis.

Objective To compare the efficacy and safety ofetanercept injection 50 mg once weeklycombined with methotrexate (MTX) therapy for patients withactive rheumatoid arthritis.Methods This studyconsists of 2 parts：a 12-week double-blind treatmentperiod (part A) followed by a 12-week open-labelsafety study period (part B).The randomization oftreatments in double-blind treatment period was completedthrough the clinical operations randomization environment(CORE) system.During part A,the subjects wererandomly assigned to the etanercept 50 mg or placebo group. The dosage regimen for etanercept was 50 mgadministered subcutaneously once weekly while MTX wasadministered orally.All subjects who completed partA received 50 mg etanercept once weekly and MTX1 during theopen-label treatment.The primary endpointwas ACR 20 response at week 12.Secondary endpoint variablesincluded physician／patient global assessmentsof disease activities,duration of morning stiffness,painvisual analog scale (VAS),health assessment questi onnaire (HAQ),CRP level and tender and swollen joint counts .The results of safety between the two groupswere compared.The primary endpoint and other secondarybinary endpoints were analyzed using the Fisher’sexact test.For continuous endpoints.the change frombaseline was analyzed with analysis of covariance.Results One hundred and fifty six subjects satisfiedmodified intent-to-treat (mITT) population were enrolled during part A,of which 77 subjects were in theetanercept+MTX group,and 79 subjects were in theplacebo+MTX group respectively.A total of 149 subjectscompleted part A.As early as week 4.the ACR 20response achieved 39％ (30,77) in the etanerceptgroup,which was significantly higher than that of theplacebogroup [16％(13／79),P<0.001].At week 12,the ACR 20respouse achieved 62％(48,77)in the etanercept group and 23％(18／79) in the placebo group (P<0.01).Fromweek 4,other study endpoints including physician global assessment,patient globalassessment,duration of morning stiffness,painVAS,HAQ,CRPlevel,tender joint counts,swollen joint counts were alsocompared.The results showed that all above efficacyendpoints in the etanercept+MTX group were better than thoseof the placebo+MTX group(P<0.01).Butthere Was no significant difference in the total adverseeriects between the two groups.ConclusionEtanercept 50 mg once weekly + MTX treatment for 24 weeks iswell tolerated.During the first 12-weektreatment period,the etanercept group has shown a rapidefficacy onset and a significantly better therapeuticeffect compared to that of the placebo group.

Objective To evaluate the short-term efficacy and safety of etanercept treatment in Chinese patients with active ankylosing spondylitis ( AS ). Methods This was a 12-week multicenter,double-blind, placebo-controlled, randomized phase Ⅲ clinical study. The first part was a 6-week placebocontrolled period followed by a 6-week open-label period. The primary efficacy endpoint was the percentage of subjects achieving a 20% improvement in assessment in ankylosing spondylitis (ASAS) ( ASAS 20). The secondary efficacy endpoints were the percentage of patients achieving a 40% improvement in ASAS (ASAS 40), achieving a 50% improvement in ASAS( ASAS 50), achieving a 70% improvement in ASAS (ASAS 70), and ASAS 5/6 responses at all visits, and the improvement in subject global assessment,physician global assessment, nocturnal and total back pain, bath AS functional index ( BASFI ), bath AS disease activity index (BASDAI), spinal mobility, joint assessment and quality of life assessment. All subjects in the study were evaluated for safety. Results The primary endpoint, ASAS 20 at week 6, was achieved by 86. 5% (64/74) patients in the etanercept group compared to 29. 5% (23/78) patients in the placebo group(P ＜0. 001 ). As early as week 2, the percentages of patients achieving the ASAS 20 between the two groups were significantly different. Furthermore, the majority of secondary efficacy end points were also significantly improved. Most of adverse events (AE) were mild in nature, the commonest adverse events were elevated liver function levels, injection site reactions and nasopharyngitis. No death or serious AE were observed. Conclusion Etanercept can improve symptoms fastly,significantly and safely in Chinese patients with active AS.

Objective To investigate the mRNA expression of IKB kinase (IKK-α) and interferon-α (IFN-α) in the peripheral blood leukocytes of patients with systemic lupus erythematosus (SLE), and to explore the role of IKK-α in the production of IFN-α in SLE patients. Methods SYBR green dye I based real-time quantitative PCR was used to detect the mRNA expression levels of IKK-α and IFN-α in the peripheral blood leucocytes of SLE patients and healthy controls. Serum levels of IFN-α were measured with ELISA method. Results IKK-α mRNA expression levels in SLE patients were significantly higher than those of normal controls (P<0.05). IKK-α mRNA expression levels in SLE patients with active disease were significantly higher than patients with stable disease (P<0.01). IFN-α mRNA expression level in SLE patients was significantly lower than that of the normal controls (P<0.01). IFN-α mRNA expression levels in SLE patients with active disease were significantly higher than patients with stable disease (P<0.01). Serum levels of IFN-α in SLE patients with active disease was significantly higher than that of the normal controls and patients with stable disease (P<0.05). The anti-dsDNA antibody correlated positively, and complement C3 correlated negatively with serum concentration of IFN-α. IKK-α mRNA expression levels in SLE patients correlated positively with serum concentration of IFN-α. Conclusion IKK-α correlates positively with serum concentration of IFN-α. The IFN-α level is significantly correlated with disease activity, This suggests that IKK-α may play an important role in the pathogenesis of SLE.

Objective To study 6 type Ⅰ interferon (IFN)-inducible genes (IFIT4, IFI44, Ly6e,OAS1, OAS2 and OAS3) in patients with lupus nephritis (LN) and analyze its correlated expression levels with disease activity and/or clinical manifestations. Methods Total RNA was obtained simultaneously from kidney tissues and peripheral blood cells of 12 patients with diffuse proliferative lupus nephritis and 10 normal controls. Moreover, peripheral blood cells were obtained from 119 LN patients and 35 normal controls. Total RNA was extracted and reversely transcribed into complementary DNA. Gene expression levels were measured by real-time polymerase chain reaction by comparing to a housekeeping gene, and IFN score was calculated. Disease activity was determined by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Results The 6 genes were highly expressed in diffuse proliferative lupus nephritis patients compared with normal controls. IFN scores were positively correlated with SLEDAI score, the concurrent presences of anti-double-stranded DNA (anti-dsDNA) antibodies (P<0.05) and hypocomplementemia (P<0.01). Conclusion The 6 IFN-inducible genes are highly expressed iri LN patients. IFN scores are elevated in active lupus nephritis patients, in patients with positive anti-ds-DNA antibody and hypocomplementemia. IFN scores may be a useful biomarker for lupus nephritis therapy.

Objective To evaluate the efficacy of medical supervised aerobic exercise on physical activity, quality of life and psychological status in patients with systemic lupus erythematosus (SLE). Methods SLE patients who fulfilled ACR criteria were recruited and divided into 2 groups: exercise group (n=24) and control group (n=25). The patients in the exercise group were supervised to have aerobic exercises. The intensiveness of exercise was determined by 20%-40% of maximum heart rate reservation. Visual analog scale (VAS), SLE disease activity index (SLEDAI) score, physical working capacity (PWC170), SF-36 and profile of mood states (POMS) of the two groups were used to evaluate the changes at the baseline, 1 month, 3 months and 6 months after this study. Results The 2 groups were homogeneous and comparable in disease activity at baseline. 1, 3 and 6 months after the study, the VAS, PWC170, POMS and SF -36 of SLE patients were improved in certain degrees in both groups, while the improvement of VAS (P<0.05), PWCITO (P< 0.01 ) and social function of SF-36 (P<0.05) of exercise group were significantly more evident than those of the control group in 6 months after study without any impact on disease condition. There was a high negative correlation between VAS and 5 categories of POMS (r=-6.26~-0.393, P<0.01 ) and a more relevant positive association between VAS and 2 categories of POMS (r=0.534~0.611, P<0.01). Conclusion The data demonstrate that the supervised aerobic exercise can ameliorate physical capacity, improve quality of life and psychological and emotional status in the state SLE patients without aggravating disease per se.

Objective To explore the pathogenic genes relevant to Behcet's disease (BD) by building the differentail gene expression profiles of peripheral blood leukocytes in BD. Methods Oligonucleotide gene array from Affymetrix Company was applied to study the differed expression levels of whole genome between three age and sex matched BD patients and normal controls. Four genes, BCL6, LRAP, ICOSLG and MME, were selected to be tested for gene expression levels by real-time PCR in the groups of BD, normol controls (NC), Lupus and rheumatoid arthritis (RA) peticnts. Results ① Differential gene expression profile of BD compared to that of normal controls was built up. It contained 89 up-regulated and 57 down-regulated genes. ② Four genes mentioned above had significantly higher expression levels in active BD patients than those in NC but had lower exoression levels in stable BD patients. The expression levels of BCL6 and MME were also proved to be increased significantly in BD than in RA and SLE patients. Conclusion ① Our work shed some light on further research of the etiopathogenesis of BD. ② The expression levels of the four genes are proved to be relevant to BD the first time by us. Further analysis showes that TNF-α and IFN-γ can up-regulate the expression levels of BCL6, LRAP and ICOSLG which may be novel to BD. The MME gene is expressed on the surface of cells, which is convenient for test and may potentially be a marker for the diagnosis of BD.

Objective To explore the role of chemokines and ehemokine receptors in the etiopathog-enesis of diffuse proliferative lupus nephritis (LN). Methods ① Total RNA from the kidney tissues and peripheral blood cells of 12 patients with diffuse proliferative LN and 10 normal controls were prepared simultaneously and reverse transcribed into complementary DNA. Sybr green dye based real-time quantitative PCR method was used to compare the expression levels (indicated as-AACt value) of MCP-1, CCL19,CXCLg, CXCL10 and CCR2, CCR7, CXCR3. ② Immunofluoresceee labeling and immunohistochemical staining technique were used to observe the distribution of chemokines MCP-1, CCL19, CXCL9 and CXCL10 in normal and patients kidney tissues. Results The 4 chemokines genes (MCP-1, CCL19, CXCL9 and CXCL10) were consistently highly expressed in kidney tissues and peripheral blood ceils of diffuse proliferative LN patients compared with normal controls. The 2 chemokine receptors, CCR2 and CXCR3 were also overexpressed in peripheral blood cells of diffuse proliferative LN patients. There was nearly no expression of these 4 chemokine proteins in normal kidneys. But they were found in glomeruli of diffuse proliferative LN patients. Conclusion The expression of chemokines in the peripheral blood cells may be used as biomarkers for LN. Further study maybe lead to the development of specific drugs targeting at them for the treatment of systemic lupus erythematosus (SLE).

OBJECTIVETo develop an improved substrate for indirect immunofluorescence test (IIF) for detecting anti-Ro60/Sjogren's syndrome A (Ro/SSA) autoantibodies.

METHODS60-kDa Ro/SSA autoantigens (Ro60) cDNAs were obtained from human placental cDNA library using polymerase chain reaction (PCR) and were cloned into the mammalian expression vector-pEGFP-C1. Then, the recombinant plasmids were transfected into HEp-2 cells. We confirmed the overexpression, localization and antigenicity of fusion proteins in transfected cells by means of immunoblotting, confocal fluorescence microscopy and IIF. HEp-2 and HEp-Ro60 were analyzed by IIF using a panel of 10 precipitin-positive anti-Ro human sera simultaneously.

RESULTSStable expression of Ro60-green fluorescent protein (Ro60-GFP) fusion proteins were maintained ten more generations. Ro60-GFP kept the antigenicity of Ro while demonstrating its own characteristic immunofluorescent pattern in HEp-Ro60 cells. The transfectants dramatically increased the sensitivity of IIF testing (a mean increase of 6.7-fold in endpoint titer). Eight over ten (8/10) positive anti-Ro sera showed characteristic immunofluorescent patterns for HEp-Ro60, including two sera that were anti-nuclear antibodies (ANA) negative for untransfected HEp-2. IIF-ANA in all healthy sera was negative for HEp-Ro60.

CONCLUSIONSAs a new substrate for IIF, the Ro60 transfectants can be used to detect anti-Ro antibodies. In addition, transfected HEp-2 cells keep the immunofluorescent properties of HEp-2 cells in IIF-ANA tests and can be employed as a substrate for routine IIF-ANA detection.

Antibodies, Antinuclear ; blood ; Autoantigens ; Cell Line ; Fluorescent Antibody Technique, Indirect ; Humans ; Molecular Weight ; RNA, Small Cytoplasmic ; Recombinant Fusion Proteins ; immunology ; Ribonucleoproteins ; immunology ; Transfection

OBJECTIVETo evaluate the efficacy and safety of leflunomide in comparison with methotrexate (MTX) on patients with rheumatoid arthritis (RA) in China.

METHODSFive hundred and sixty-six patients with active rheumatoid arthritis were randomly assigned to receive leflunomide at 20 mg once daily or MTX at 15 mg once weekly in a controlled trial. Five hundred and four patients completed the 12-week treatment and some patients continued the treatment for 24 weeks.

RESULTSBoth leflunomide and MTX could improve the symptoms, signs, and joint function, but there were no changes in X-ray observations of patients with rheumatoid arthritis. In the leflunomide group, the overall rates of effectiveness at 12 weeks and 24 weeks were 86.94% and 92.31% respectively; the rates of remarkable improvement were 64.95% and 79.81% respectively. In the MTX group, the overall rates of effectiveness at 12 weeks and 24 weeks were 84.04% and 83.15% respectively; the rates of remarkable improvement were 56.81% and 75.28% respectively. According to intent-to-treat analysis, the ACR 20% response rates at 12 weeks and 24 weeks in the leflunomide group were 62.54% and 67.18% respectively, compared with 60.08% and 61.32% respectively in MTX group. No statistical differences were shown in the efficacy between the two groups (P > 0.05). The adverse events in the leflunomide group were gastrointestinal symptoms, skin rash, alopecia, nervous system symptoms, decreased leukocyte count, and elevation of alanine aminotransferase (ALT). Most of these side effects were mild and transient. The incidence of adverse events in the leflunomide group was 16.84%, significantly lower than that in MTX group (28.17%, P = 0.002).

CONCLUSIONSLeflunomide is effective in the treatment of RA with less adverse events than MTX. Its efficacy is similar to MTX, but the incidence of adverse events and the rate of withdrawal due to adverse events were lower in the leflunomide group than in MTX group.

Antirheumatic Agents ; adverse effects ; therapeutic use ; Arthritis, Rheumatoid ; drug therapy ; Female ; Growth Inhibitors ; adverse effects ; therapeutic use ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Isoxazoles ; adverse effects ; therapeutic use ; Male ; Methotrexate ; adverse effects ; therapeutic use ; Middle Aged