明胶酶-B及其抑制剂与类风湿性关节炎

明胶酶 B(GelatinaseB)属于金属蛋白酶 (MMPs)家族 ,又称MMP - 9,其主要功能是降解细胞外基质中的Ⅳ型、Ⅴ型胶原和明胶。在体内可参与多种生理、病理过程。明胶酶 -B可在多个水平上受多种方式的调节 ,基因表达的调节、酶原激活的调节、酶抑制剂的调节。明胶酶 B在类风湿性关节炎 (RA)发生发展过程中起多种不同的作用 ,包括对骨和软骨的破坏、对滑膜新血管形成的影响和对细胞因子的调节。由于明胶酶 -B在RA发病过程中起重要作用 ,因此抑制其活性是对RA进行治疗的一种有效手段。     

Elevated Levels of Serum IgA against Saccharomyces cerevisiae Mannan in Patients with Rheumatoid Arthritis

This study was undertaken to investigate whether levels of anti-Saccharomyces cerevisiae mannan antibodies （ASCMA） in human sera, a marker for several autoimmune diseases, correlate with rheumatoid arthritis （RA）. ASCMA-IgA, -IgG and -IgM levels were measured with enzyme linked immunosorbent assays （ELISA） in patients with RA （n = 30） and 152 healthy adult controls. ASCMA-IgA prevalence was significantly higher in RA patients （40%） than in healthy subjects （5.3%）. A strong correlation between levels of ASCMA-IgA and CRP （r = 0.695; p 〈 0.01） and ESR （r = 0.708; p 〈 0.01） in RA patients was observed. No significant differences in ASCMA-IgG or IgM levels were noted between RA patients and healthy control subjects in the present study. This result differs from previous reports. It remains to be evaluated whether elevated ASCMA-levels are common to all rheumatic disorders.     

Serum Pro-hepcidin Levels in Rheumatoid Arthritis and Systemic Lupus Erythematosus

Hepcidin is a principal iron regulatory hormone and its expression is stimulated by cytokines. The aim of this study was to determine serum levels of the prohormone form of hepcidin, pro-hepcidin, in rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). The study included 72 RA and 28 SLE patients and 33 healthy controls (HC). Serum iron status, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6 and pro-hepcidin levels were determined. Pro-hepcidin levels in the RA group was higher than SLE and HC groups (p < 0.05, p < 0.001, respectively). Pro-hepcidin levels did not correlate with disease activity scores, cytokine levels and serum iron status in the RA and SLE groups, while it correlated with TNF-alpha, IL-6 and ferritin levels in the HC group (r = 0.459, p < 0.01, r = 0.374, p < 0.05, r = -0.603, p < 0.01, respectively). Pro-hepcidin levels show extremely wide variations within the groups as do iron status and cytokines. Despite these wide variations correlation analysis do not reveal anything.     

Increased Serum GP88 (Progranulin) Concentrations in Rheumatoid Arthritis

GP88 (Progranulin; PGRN) is a secreted glycosylated protein with important functions in several processes, including immune response and cancer growth. Recent reports have shown that PGRN is a therapeutic target for rheumatoid arthritis (RA) because of its capability to bind with tumor necrosis factor receptor (TNFR). However, the serum PGRN level in RA patients has not been investigated. We used enzyme-linked immunosorbent assay (ELISA) to quantify the serum levels of PGRN in 417 healthy subjects, 56 patients with RA and 31 patients with osteoarthritis (OA). In RA patients, we also measured the serum TNF-α and sTNFR concentration. Immunohistochemical staining of PGRN was performed using synovectomy tissue of RA patients. The serum PGRN normal range was established as 40.1?±?8.7 ng/ml. PGRN levels were not influenced by sex or age. A significant increase in serum PGRN levels was observed in RA (50.2?±?11.1 ng/ml) and OA (45.4?±?6.6 ng/ml) groups compared to those in age-matched healthy controls (40.4?±?9.9 ng/ml) (p?TNF-α and sTNFR 2 concentration. Furthermore, PGRN/TNF-α ratio was correlated the stage of the disease in RA patients. The concentrations of serum PGRN in RA were found to be significantly higher than those in age-matched healthy controls, although it remains to be clarified how blood PGRN is related to the pathogenesis of RA. Our results showed that the serum PGRN may be a useful approach to monitor the disease activity in RA patients.     

Autoantibodies to Citrullinated (Poly)Peptides: A Key Diagnostic and Prognostic Marker for Rheumatoid Arthritis

Hepatitis B infection is one of the most important causes of acute and chronic liver disease. During the 1980s, genetically engineered hepatitis B vaccines (HBVs) were introduced in the United States. A large-series of serious autoimmune conditions have been reported following HBVs, despite the fact that HBVs have been reported to be “generally well-tolerated.” A case-control epidemiological study was conducted to evaluate serious autoimmune adverse events prospectively reported to the vaccine adverse events reporting system (VAERS) database following HBVs, in comparison to an age, sex, and vaccine year matched unexposed tetanus-containing vaccine (TCV) group for conditions that have been previously identified on an a priori basis from case-reports. Adults receiving HBV had significantly increased odds ratios (OR) for multiple sclerosis (OR = 5.2, p < 0.0003, 95% Confidence Interval (CI) = 1.9 ? 20), optic neuritis (OR = 14, p < 0.0002, 95% CI = 2.3 ? 560), vasculitis (OR = 2.6, p < 0.04, 95% CI = 1.03 ? 8.7), arthritis (OR = 2.01, p < 0.0003, 95% CI = 1.3 ? 3.1), alopecia (OR = 7.2, p < 0.0001, 95% CI = 3.2 ? 20), lupus erythematosus (OR = 9.1, p < 0.0001, 95% CI = 2.3 ? 76), rheumatoid arthritis (OR = 18, p < 0.0001, 95% CI = 3.1 ? 740), and thrombocytopenia (OR = 2.3, p < 0.04, 95% CI = 1.02 ? 6.2) in comparison to the TCV group. Minimal confounding or systematic error was observed. Despite the negative findings of the present study regarding the rare serious adverse effects of HBVs, it is clear that HBV does, indeed, offer significant benefits, but it is also clear that chances of exposure to hepatitis B virus in adults is largely life-style dependent. Adults should make an informed consent decision, weighing the risks and benefits of HBV, as to whether or not to be immunized.     

血清胞苷脱氨酶在活动性类风湿关节炎诊疗中的意义

目的:探讨测定血清胞苷脱氨酶(CD)含量对活动性类风湿关节炎(RA)患者的诊疗价值。方法:采用分光光度法测定血清中的胞苷脱氨酶活性,同时用免疫比浊法测定C反应蛋白(CRP),魏氏法测定血沉(ESR)。结果:活动性RA组血清CD含量(14.80±2.11)U/m l,显著高于正常对照组(4.86±1.86)U/m l,(P<0.01);CRP(51.46±20.43)mg/L和ESR(85.03±27.6)mm/1h明显高于正常对照组(3.40±2.21)mg/L,(13.04±4.89)mm/1h(均P<0.01);活动性RA患者组CD含量与ESR、CRP成直线关系,相关系数r分别为0.6324(P<0.01)与0.8013(P<0.01)。结论:血清CD活性可作为活动性RA患者急性感染的早期诊断指标,对活动性RA患者的预后判断亦有临床价值。     

Serum Levels of Calreticulin in Correlation with Disease Activity in Patients with Rheumatoid Arthritis

Objective

The aim of our study was to investigate the contribution of serum calreticulin (CRT) in the assessment of disease activity in rheumatoid arthritis (RA).

Methods

Serum CRT levels were measured by ELISA in 70 patients with established RA, 30 systemic lupus erythematosus (SLE), 25 other autoimmune diseases, 20 osteoarthritis (OA), and 35 of healthy controls (HC). Correlations of CRT serum levels with disease activity [Disease Activity Score for 28 joints (DAS28)], erythrocyte sedimentation rate(ESR) and C-reactive protein (CRP) were assessed. Serum CRT levels were also detected in RA patients whose RF, anti-CCP and anti- MCV antibodies were positive and negative.

Results

Serum CRT levels in RA patients (4.817?±?2.425 ng/ml) was significantly higher (P <0.05) compared with those in the serum of OA (3.574?±?0.942 ng/ml), SLE (4.013?±?1.536 ng/ml), other autoimmune diseases (3.882?±?0.837 ng/ml) and HC (3.726?±?0.627 ng/ml). Significant positive correlation of CRT with DAS28, ESR and CRP was found in RA patients. Furthermore, RA patients whose anti-CCP and anti-MCV antibodies were positive had higher levels of CRT (P?<?0.01).

Conclusion

Serum CRT levels were increased in patients with RA compared with those controls. Moreover, a significant correlation was observed between serum CRT levels and disease activity in RA. It might be used as a potential biomarker for clinical diagnosis and provide additional information regarding disease activity along with the traditional indices such as ESR and CRP.     

IL‐12B Gene Polymorphisms and IL‐12 p70 Serum Levels Among Patients with Rheumatoid Arthritis

Rheumatoid arthritis (RA) is one of the autoimmune diseases, where different polymorphisms in cytokine genes play a pathogenic role. IL‐12 is now recognized as a critical cytokine in terms of regulating the balance between Th1 and Th2 cells. We investigated the role of single nucleotide polymorphisms (SNPs) (rs3212227 (A/C) and rs17860508 (CTCTAA/GC)) of the IL‐12B gene in the genetic susceptibility to RA and in the severity of the disease. Six hundred and thirty‐four Caucasian RA patients and 341 healthy matched controls were studied using PCR‐RFLP method and high‐resolution melting analysis. Concentration of IL‐12 cytokine level in serum was evaluated using ELISA. The genotype frequency did not deviate from HWE in each examined group. Frequencies of the rs3212227 CC genotype were statistically higher in patients with RA compared with the healthy control group in both codominant and recessive models (P = 0.037; P = 0.04, respectively). The frequency of rs3212227 C allele also showed similar tendency (P = 0.07). IL‐12 level in serum was significantly higher in RA group compared with control (P < 0.0001). We observed that increased IL‐12 serum level was correlated with higher number of tender and swollen joints, ExRA presence and higher levels of haemoglobin, CRP and PLT. Also higher IL‐12 level in serum was observed within RA patients with hypertension. Present findings indicated that IL‐12p40 + 1188A/C polymorphism as well as IL‐12p70 protein levels may be associated with RA in the Polish population.     

血清MMP-3、TIMP-1及HC gp-39在早期RA患者的表达与临床的相关性研究

目的：研究血清基质金属蛋白酶-3（matrix metalloproteinase 3,MMP-3）、金属蛋白酶组织抑制剂-1（tissue inhibitors of metalloproteinase 1,TIMP-1）及人类软骨糖蛋白-39（human cartilage glycoprotein 39,HC gp-39）在早期类风湿关节炎（rheumatoid arthritis,RA）患者的表达及其相关性因素,探讨其在早期RA的可能意义。方法：选取45例病程〈1年的早期RA患者和32例健康查体者,采用酶联免疫吸附试验（ELISA）分别测定血清MMP-3、TIMP-1及HC gp-39水平,同时测定关节肿胀数（SJC）、关节压痛数（TJC）、血沉（ESR）、C反应蛋白（CRP）、类风湿因子（RF）等相关实验室指标。结果：早期RA患者血清MMP-3、TIMP-1及HC gp-39水平明显高于正常对照组（P〈0.01）。不同X线分期各指标差异有统计学意义（P〈0.05）。不同关节功能分级时各指标差异无统计学意义（P〉0.05）。三指标与SJC（P〈0.01）、TJC（P〈0.01）、ESR（P〈0.01）、CRP（P〈0.01）呈正相关,与年龄、病程、晨僵时间、RF无明显相关（P〉0.05）。结论：MMP-3、TIMP-1及HC gp-39在早期RA患者血清中呈高水平表达,可作为反映疾病活动和早期RA患者出现骨侵蚀的有益预测指标。     

Gelatinase A (MMP-2) is necessary and sufficient for renal tubular cell epithelial-mesenchymal transformation

Progressive renal interstitial fibrosis and tubular atrophy represent the final injury pathway for all commonly encountered forms of renal disease that lead to end-stage renal failure. It has been recently recognized that myofibroblastic cells are the major contributors to the deposition of interstitial collagens. While there are several potential cellular sources of myofibroblasts, attention has focused on the transformation of the organized tubular epithelium to the myofibroblastic phenotype, a process potently driven both in vitro and in vivo by transforming growth factor-beta1 (TGF-beta1). Integrity of the underlying basal lamina provides cellular signals that maintain the epithelial phenotype, and disruption by discrete proteases could potentially initiate the transformation process. We demonstrate that TGF-beta1 coordinately stimulates the synthesis of a specific matrix metalloproteinase, gelatinase A (MMP-2), and its activator protease, MT1-MMP (MMP-14), and that active gelatinase A is absolutely required for epithelial-mesenchymal transformation induced by TGF-beta1. In addition, purified active gelatinase A alone is sufficient to induce epithelial-mesenchymal transformation in the absence of exogenous TGF-beta1. Gelatinase A may also mediate epithelial-mesenchymal transformation in a paracrine manner through the proteolytic generation of active TGF-beta1 peptide. MT1-MMP and gelatinase A were co-localized to sites of active epithelial-mesenchymal transformation and basal lamina disruption in the rat remnant kidney model of progressive renal fibrosis. These studies indicate that a discrete matrix metalloproteinase, gelatinase A, is capable of inducing the complex genetic rearrangements that characterize renal tubular epithelial-mesenchymal transformation.     

抗CCP抗体和RF联检在RA诊疗中的临床价值

目的：研究抗环瓜氨酸肽（anti-cyclic citrullinated peptide,Anti-CCP）（抗CCP抗体）和RF的检测在类风湿关节炎（RA）诊疗中的临床价值。方法：分别用酶联免疫吸附试验（ELISA）、BeckMan全自动蛋白分析仪同时检测早期RA组（病程〈1年）42例,RA组（病程〉1年）40例,非RA对照组40例患者血清抗CCP抗体和RF。结果：早期RA组、RA组的抗CCP抗体、RF阳性率显著高于非RA对照组（P〈0.05）RA组抗CCP水平显著高于早期RA组（P〈0.01）,两者RF无显著差别（P〉0.05）。RA组与早期RA组CCP抗体与RF二者无相关性。结论：联检抗CCP抗体、RF有助于类风湿的早期诊断和预测病情的进展。     

Gelatinase B (MMP-9) is not essential in the normal kidney and does not influence progression of renal disease in a mouse model of Alport syndrome

Matrix metalloproteinases are matrix degrading enzymes implicated in many biological processes, including development and inflammation. Gelatinase B (gelB; also known as MMP-9) is expressed in the kidney and is hypothesized to be involved in basement membrane remodeling and in preventing pathogenic accumulation of extracellular matrix in the kidney. Inhibition of gelB activity in metanephric organ culture disrupts branching morphogenesis of the ureteric bud, suggesting that gelB plays a role in kidney development in vivo. We studied kidneys of gelB-deficient mice to search for developmental, histological, molecular, ultrastructural, and functional defects. Surprisingly, no differences between gelB-/- and control kidneys were detected, and renal function was normal in gelB mutants. In addition, gelB-/- embryonic kidneys developed normally in organ culture. Gelatinase B-deficient mice were bred with Col4a3-/- mice, a model for Alport syndrome, to determine whether gelB influences the progression of glomerulonephritis. This is an important question, as it has been hypothesized that proteases are involved in damaging Alport glomerular basement membrane. However, the presence or absence of gelB did not affect the rate of progression of renal disease. Thus, gelB does not have a discernible role in the normal kidney and gelB is not involved in the progression of glomerulonephritis in a mouse model of Alport syndrome.     

Expression of ICAM1 and VCAM1 Serum Levels in Rheumatoid Arthritis Clinical Activity. Association with Genetic Polymorphisms

To investigate the association of sICAM-1 and sVCAM-1 with ICAM1 721G>A and VCAM1 1238G>C polymorphisms and rheumatoid arthritis (RA) clinical activity, sixty RA patients and 60 healthy non-related subjects (HS) matched for age and sex were recruited. Soluble adhesion molecules were determined by ELISA technique. Rheumatoid factor (RF), C reactive protein (CRP) and the erythrocyte sedimentation rate (ESR) were measured by routine methods. Disability and clinical activity was measured with Spanish-HAQ-DI and DAS28 scores, respectively. The ICAM1 and VCAM1 polymorphism were identified using the PCR-RFLP procedure. Inter-group comparison showed increased levels of sICAM-1 and sVCAM-1 in RA patients (284 and 481 ng/mL) versus HS (132 and 280 ng/mL); in the RA group, significant correlations between sVCAM-1 and RF (r = 0.402), ESR (r = 0.426), Spanish-HAQ-DI (r = 0.276), and DAS28 (r = 0.342) were found, whereas sICAM-1 only correlated with RF (r = 0.445). In RA patients, a significant association with the 721A allele of ICAM1 polymorphism (p = 0.04), was found. In addition, the allele impact (G/A + A/A) of this polymorphism was confirmed, (p = 0.038, OR = 2.3, C.I. 1.1–5.0). sVCAM-1 and sICAM-1 serum levels reflected the clinical status in RA, independently of the ICAM1 and VCAM1 polymorphism. However, the ICAM1 721A allele could be a genetic marker to RA susceptibility.     

类风湿性关节炎患者IL-6、IL-18和CRP的水平变化及意义

目的：研究类风湿性关节炎（RA）患者血清白细胞介素-6（IL-6）、白细胞介素-18（IL-18）和C-反应蛋白（CRP）水平的变化及其临床意义。方法：收集84例RA患者,以70例健康体检者作对照。采用双抗体夹心酶联免疫吸附法测定血清IL-6、IL-18和免疫荧光法测定CRP的水平,并测定血小板计数（Plt）、血沉（ESR）、类风湿因子（RF）。结果：RA患者的血清Plt、ESR、RF、IL-6、IL-18和CRP的含量明显高于健康对照组（P〈0.01）。RA患者活动期上述指标含量（除RF外）均显著高于稳定期（P〈0.01）,Plt升高RA患者组与Plt正常组相比,RF、ESR、IL-6、IL-18和CRP水平均有明显统计学差异（P〈0.05）。结论：IL-6、IL-18和CRP在RA患者的疾病发展过程中发挥着重要作用,它们的水平变化与RA患者病情有关,联合动态监测有助于临床观察RA患者的病情变化和治疗效果。     

Structural alterations of the vascular wall in magnesium-deficient mice. A possible role of gelatinases A (MMP-2) and B (MMP-9).

Vascular alterations during magnesium deficiency have long been known but the implicated mechanisms have so far been poorly documented. In this preliminary assay, we compared the thoracic aortic histology in Swiss OF1 mice fed a severe magnesium-deficient diet (50 +/- 5 ppm) for 42 days to that of controls fed a standard diet (1700 +/- 100 ppm magnesium). It appeared (eosin-haematoxylin coloration) that, in magnesium-deficient mice, the aortic wall was thinner than in controls. Specific colorations of the two of main fibers vascular tissue (collagens and elastin) showed severe structural alterations of both components. These changes were consecutive to the expression of matrix metalloproteinases (MMP) -2 and -9 which were present as zymogens (inactive forms) in controls and supposed to be present in their active and inactive forms in magnesium-deficient mice (zymography). These changes which have not been reported so far would explain, at least in part, the sensitivity of magnesium-deficient mice to various stress or xenobiotics.     

类风湿关节炎患者骨密度改变与血清25-羟维生素D3水平的关系分析

目的 分析类风湿关节炎(RA)患者骨密度改变与血清25-羟维生素D3[(25-(OH)D3)]水平的关系.方法 选取2014年5月至2017年5月我院收治的RA患者96例为研究对象,另选取50例同期入院体检的健康志愿者为对照组,对比不同严重程度RA患者与对照组血清25-(OH)D3水平及骨密度(BMD)、骨密度T值,分析25-(OH)D3不同缺乏程度患者BMD、骨密度T值,分析RA患者25-(OH)D3水平与BMD及骨密度T值关系.结果 RA组血清25-(OH)D3水平、BMD、骨密度T值低于对照组(P<0.05);RA分期Ⅳ期患者血清25-(OH)D3水平、BMD、骨密度T值低于Ⅰ、Ⅱ、Ⅲ期(P<0.05);随25-(OH)D3缺乏程度加重,RA患者BMD、骨密度T值下降(P<0.05);相关分析显示RA患者血清25-(OH)D3水平与BMD、骨密度T值呈正相关(r=0.453、0.324,P均<0.05).结论 RA患者骨密度改变与其血25-(OH)D3水平存在密切关系,随RA病情加重,患者血清25-(OH)D3水平及BMD下降,且25-(OH)D3水平缺乏程度加重,骨密度降低越明显,因此在RA治疗中可补充维生素D而改善预后.     

Matrix metalloproteinase-9 (MMP-9) in allergic nasal polyps

Matrix metalloproteinases (MMPs) are known to play important roles in the invasion of nasal mucosa by inflammatory cells through degradation of extracellular matrix. Matrix metalloproteinase-9 (MMP-9) is considered to play a role in the pathogenesis of nasal polyp. The aim of the present study was to compare plasma MMP-9 levels of patients with nasal polyp of different etiologic origins, those with allergic nasal polyp (ANP) and non-allergic nasal polyp (NANP). In all, 29 patients (20 NANP and 9 ANP) and 20 healthy subjects were included in this study. Plasma MMP-9 levels were measured using ELISA. Plasma MMP-9 levels were higher (p<0.05) in ANP patients than in NANP patients and controls. In the group with ANP, plasma MMP-9 levels showed a positive correlation (p<0.05) with eosinophil counts. Plasma MMP-9 levels of NANP patients and control groups were similar (p>0.05). Moreover, that the highest levels of MMP-9 were in ANP patients may indicate the usefulness of this parameter in differentiating between the different etiologic origins of nasal polyp. Further studies are required to elaborate on the relationship between MMP-9 levels and allergic reactions.