Soluble receptor for advanced glycation end products in multiple sclerosis: a potential marker of disease severity

OBJECTIVES: To compare serum levels of the receptor for advanced glycation end products (sRAGE) between multiple sclerosis (MS) patients and healthy control subjects, and to investigate whether serum sRAGE levels correlate with MS disease severity as indicated by the Kurtzke Expanded Disability Status Scale (EDSS). METHOD: 37 patients with clinical diagnosis of MS and 22 healthy control subjects were investigated in a cross-sectional study using enzyme-linked immunosorbent assays (ELISA). RESULTS: Serum levels of sRAGE were found to be significantly lower in MS patients compared to levels in healthy controls (p = 0.005). A trend toward lower levels of serum sRAGE was observed in female MS patients compared to their male counterparts (p = 0.05). A relationship between sRAGE and EDSS, and sRAGE and rate of clinical relapse was observed (p = 0.012). CONCLUSION: The significant reduction of sRAGE in MS patients relative to healthy controls supports the potential role for RAGE axis in MS clinical pathology. Lower levels of sRAGE may be associated with enhanced inflammatory responses. Based on these observations, further investigations into the role of sRAGE in MS clinical pathology is warranted.     

MicroRNA Expression Aberration in Chinese Patients with Relapsing Remitting Multiple Sclerosis

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease that is caused by an abnormal immune system that attacks the central nervous system. MicroRNAs (miRNAs) are known to play essential roles in the immune system. Most of the previous studies about miRNA dysregulation in MS have focused on European populations. In the present study, the miRNA expression profiles associated with Chinese MS patients are investigated. Here, human miRNA expression profiling experiments were performed on the peripheral blood mononuclear leukocytes from ten MS patients and ten healthy controls. Nine overexpressed and six underexpressed miRNAs were found. The 15 miRNAs were validated independently in a second cohort of 40 MS patients and 40 controls by real-time PCR; six miRNAs were significantly abnormally expressed, and principal component analysis of six miRNAs indicated that the MS patients could be clearly differentiated from the healthy controls based on the miRNA expression patterns. This study provided the indications of abnormal miRNA expression patterns in the peripheral blood mononuclear leukocytes of Chinese MS patients. The potential roles of these differentially expressed miRNAs as MS biomarker and in pathogenesis need to be investigated in future studies.     

强直性脊柱炎患者外周血可溶性CD146的表达及意义

背景：研究发现在炎症因子的刺激下内皮细胞和活化的T细胞表面表达CD146显著增加,故推测CD146可能参与了组织的炎症反应。 目的：观察强直性脊柱炎患者外周血可溶性CD146的表达水平和临床意义。 方法：选择上海交通大学附属第六人民医院住院的强直性脊柱炎患者62例,分为两组。活动期组46例,非活动组16例。同时选择同期医院健康职工和大学生20名为对照组。在强直性脊柱炎患者初诊时评估BASDAI、BASFI、强直性脊柱炎患者总体评分(PGA)、夜间痛、总体背痛评分、晨僵时间并测定红细胞沉降率、C-反应蛋白水平。应用ELISA法检测62例强直性脊柱炎患者与20例正常人外周血可溶性CD146的表达水平、魏氏法测定血沉及免疫比浊法测定C-反应蛋白。对强直性脊柱炎患者可溶性CD146与各项检测指标相关分析。 结果与结论：强直性脊柱炎患者的外周血清可溶性CD146的表达水平较正常对照组显著增高(P < 0.05);活动期强直性脊柱炎患者可溶性CD146的表达水平较非活动期和正常对照组显著增高(P < 0.05);强直性脊柱炎患者血清可溶性CD146的表达水平与BASDAI 呈正相关(P < 0.05);累及外周关节型者血清可溶性CD146较单独中轴型强直性脊柱炎患者和正常对照组显著增高(P < 0.05)。结果表明,外周可溶性CD146表达水平的增加与强直性脊柱炎患者疾病的活动性有关,其可能参与了强直性脊柱炎的发病过程。     

Increased serum levels of soluble CD14 indicate stable multiple sclerosis

The aim of this study was to evaluate a possible role of soluble CD14 (sCD14) in multiple sclerosis (MS). We found that sCD14 serum levels measured by ELISA were higher in MS patients compared to neurological and healthy controls. Within the MS group sCD14 levels were increased in relapsing-remitting and secondary progressive MS compared to primary progressive MS. Furthermore, sCD14 concentrations were increased during stable disease. An increased expression of sCD14 was also detected after treatment with interferon-beta. In summary, we report evidence that serum sCD14 levels are increased in MS and correlate inversely with disease activity in relapsing MS patients.     

重症肌无力患者外周血microRNA-146a、炎症细胞因子的表达水平变化及其相关性分析

目的 探讨重症肌无力患者外周血microRNA-146a、炎症细胞因子（IL-6、CRP、TNF-α）的表达水平变化及相关性分析。 方法 选取2015年7月-2018年11月本院收治的重症肌无力患者58例作为试验组（MG组）,另选取同期体检健康者30例作为对照组;采集空腹外周静脉血,分离单个核细胞并提取总RNA;采用RT-PCR技术检测2组外周血单个核细胞总microRNA-146a的相对表达水平;采用ELISA技术检测2组外周炎症细胞因子（IL-6、CRP、TNF-α）的表达水平;比较2组microRNA-146a、炎症细胞因子（IL-6、CRP、TNF-α）的表达水平,分析重症肌无力患者microRNA-146a、症细胞因子（IL-6、CRP、TNF-α）表达水平与重症肌无力定量评分（QMGS）的相关性以及microRNA-146与三种炎症细胞因子表达水平的相关性。 结果 MG患者外周血microRNA-146a、IL-6、CRP、TNF-α高表达(P<0.05);MG患者外周血microRNA-146a、IL-6、CRP、TNF-α表达水平与QMGS评分呈正相关(r=0.795,0.642,0.365,0.743,P均<0.05);MG患者microRNA-146a表达水平与IL-6、TNF-α表达水平呈正相关(r=0.700,0.870,P<0.05),与CRP表达水平无相关性(r=0.235,P>0.05)。 结论 MG患者外周血存在microRNA-146a的高表达,且可能通过调控IL-6、TNF-α的表达来参与MG发病。     

Clinical and MRI disease activity in multiple sclerosis are associated with reciprocal fluctuations in serum and cerebrospinal fluid levels of soluble HLA class I molecules

The goal of our study was to clarify the contribution of soluble human leukocyte antigens class I (sHLA-I) in multiple sclerosis (MS) immune dysregulation. We retrospectively evaluated by ELISA cerebrospinal fluid (CSF) and serum sHLA-I levels in 79 relapsing-remitting (RR), 26 secondary progressive (SP) and 15 primary progressive (PP) MS patients stratified according to clinical and Magnetic Resonance Imaging (MRI) evidence of disease activity. One hundred and nine patients with other inflammatory neurological disorders (OIND), 88 with noninflammatory neurological disorders (NIND) and 82 healthy donors were used as controls. An intrathecal synthesis of sHLA-I detected by a specific index was significantly more consistent in MS than in controls, with more pronounced values in MS patients with relapses and MRI enhancing brain lesions. A decrease in serum sHLA-I concentrations was observed in MS patients with demyelinating attacks, while an increase in CSF levels of sHLA-I was found in MS patients with lesional activity on MRI scans. This association between intrathecal synthesis and reciprocal fluctuations of CSF and serum levels of sHLA-I in clinically and MRI active MS seems to suggest a potential role for CSF and serum levels of sHLA-I as a sensitive marker of immune activation taking place both intrathecally and systemically in MS.     

miRNA对多发性硬化的影响

目的探讨某些特定的miRNA是否在MS的发病过程中起着某种重要的作用,为MS的治疗提供新的治疗思路。方法依据2010年McDonald诊断标准收录临床复发期MS患者,采集其静脉血;收集健康志愿者静脉血作为对照。通过TaqMan Array Human MicmRNA A+B Cards Set v3.0微阵列检测差异性表达的miRNA,用Real time RT-PCR验证某些特定miRNA在MS患者和正常对照组中的表达情况。结果应用微阵列检测756个miRNA的异常表达,表明15个miRNA在MS与正常对照组之间存在显著性差异,其中miR-146a、miR-146b呈现高表达。进而用Real time RT-PCR验证miR-146a、miR-146b在MS患者组及正常对照组的表达情况。miR-146a在MS组及对照组中表达的2~(-△△CT)值分别为5.588±2.183,2.190±1.964,P0.01;miR-146b在MS组及对照组中表达的2~(-△△CT)值分别为2.004±0.954,0.566±0.287,P0.05。结论 miR-146a、miR-146b在MS组患者中表达明显增高,说明miR-146确实与MS的发生有着密切的关系。     

Correlation between DJ-1 levels in the cerebrospinal fluid and the progression of disabilities in multiple sclerosis patients

ObjectivesDJ-1 plays a key role in the anti-oxidative stress function. Increasing evidence supports the role of oxidative stress in the pathogenesis of multiple sclerosis (MS). The aim of this study was to investigate whether the DJ-1 levels were increased in patients with MS and to examine its association with the progression of MS.MethodsQuantitative immunoblot assays were performed to evaluate the DJ-1 level in serum and cerebrospinal fluid (CSF) collected from relapsing-remitting patients with MS (n = 29), disease controls subjects (n = 14), and healthy subjects (n = 44).ResultsNo significant difference was observed in the serum DJ-1 level among the patients with MS, disease controls, and healthy controls. However, the CSF DJ-1 levels were significantly higher in the patients with MS than in the disease control subjects (P < 0.0001). A significant positive correlation was also found between the CSF DJ-1 levels and the Multiple Sclerosis Severity Score (P < 0.005, r = 0.501).ConclusionsThese results show that the CSF DJ-1 levels are significantly increased in the CSF of patients with MS and that the CSF DJ-1 levels may be associated with the disease progression of MS. Therefore, DJ-1 possibly plays an important role in the pathogenesis of MS.     

Expression of microRNAs miR21, miR146a,and miR155 in tuberous sclerosis complex cortical tubers and their regulation in human astrocytes and SEGA‐derived cell cultures

Tuberous sclerosis complex (TSC) is a genetic disease presenting with multiple neurological symptoms including epilepsy, mental retardation, and autism. Abnormal activation of various inflammatory pathways has been observed in astrocytes in brain lesions associated with TSC. Increasing evidence supports the involvement of microRNAs in the regulation of astrocyte‐mediated inflammatory response. To study the role of inflammation‐related microRNAs in TSC, we employed real‐time PCR and in situ hybridization to characterize the expression of miR21, miR146a, and miR155 in TSC lesions (cortical tubers and subependymal giant cell astrocytomas, SEGAs). We observed an increased expression of miR21, miR146a, and miR155 in TSC tubers compared with control and perituberal brain tissue. Expression was localized in dysmorphic neurons, giant cells, and reactive astrocytes and positively correlated with IL‐1β expression. In addition, cultured human astrocytes and SEGA‐derived cell cultures were used to study the regulation of the expression of these miRNAs in response to the proinflammatory cytokine IL‐1β and to evaluate the effects of overexpression or knockdown of miR21, miR146a, and miR155 on inflammatory signaling. IL‐1β stimulation of cultured glial cells strongly induced intracellular miR21, miR146a, and miR155 expression, as well as miR146a extracellular release. IL‐1β signaling was differentially modulated by overexpression of miR155 or miR146a, which resulted in pro‐ or anti‐inflammatory effects, respectively. This study provides supportive evidence that inflammation‐related microRNAs play a role in TSC. In particular, miR146a and miR155 appear to be key players in the regulation of astrocyte‐mediated inflammatory response, with miR146a as most interesting anti‐inflammatory therapeutic candidate. GLIA 2016;64:1066–1082     

Serum levels of interleukin (IL)-18, IL-23 and IL-17 in Chinese patients with multiple sclerosis

It has been reported that cytokines play an important role in the pathogenesis of multiple sclerosis (MS). The aim of this study was to evaluate the serum levels of interleukin (IL)-18, IL-23 and IL-17 in Chinese patients with MS. We compared the serum concentrations of pro-inflammatory cytokines IL-18, IL-23 and IL-17 in 39 patients with MS and 39 healthy controls matched with sex and age. Serum cytokines were measured by FlowCytomix, a kind of cytometric bead-based assay. Correlations between the serum levels of the three cytokines and disability (expanded disability status scale, EDSS), disease duration, current age and age at onset were examined. Serum concentrations of all IL-18, IL-23 and IL-17 were significantly higher in MS patients than healthy controls. There were no significant differences of the three cytokines' levels between female and male healthy controls, while the serum IL-18 level was observed significantly higher (P=0.049) in male MS patients than female MS patients. No significant correlations were observed between any of the three cytokines' levels and EDSS, disease duration and current age. However, IL-23 was found negatively correlated with age at onset in male MS patients (r(s)=-0.775, P=0.041). Our data suggest that all IL-18, IL-23 and IL-17 may be involved in the pathogenesis of MS. However, the relationships of the three cytokines and clinical characteristics of MS need to be further investigated in the future.     

脑梗死患者血清miR-497,TNF-α表达水平变化及其临床意义

目的 探讨急性脑梗死(Acute cerebral infarction,ACI)患者血清微小RNA-497(MicroRNA-497,miR-497)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)的表达水平变化及其临床意义。方法 选取2016年1月-2019年11月本院收治的96例ACI患者,称ACI组,并选取本院同期98例体检健康者,称对照组; 采用实时荧光定量PCR(Real-time fluorescent quantitative PCR,qRT-PCR)法检测所有研究对象血清miR-497表达水平; 采用酶联免疫吸附法(Enzyme-linked immunosorbent assay,ELISA)检测所有研究对象血清肿瘤坏死因子-α水平; 评估ACI患者神经功能缺损程度、计算脑梗死体积,比较不同神经功能缺损程度/脑梗死体积的ACI患者血清miR-497、TNF-α水平; Pearson法分析ACI患者血清miR-497、TNF-α水平与神经功能缺损程度评分(National institutes of health stroke scale,NIHSS)、脑梗死体积的关系; 采用受试者工作特征曲线(Receiver operating characteristic curve,ROC)评价血清miR-497、TNF-α对ACI的诊断价值。结果 ACI组血清miR-497、TNF-α水平均明显高于对照组(P<0.05); ACI患者血清miR-497、TNF-α水平随神经功能缺损程度加重、脑梗死体积增加均呈递增趋势(P均<0.05); ACI患者血清miR-497、TNF-α水平与脑梗死体积、NIHSS评分均呈正相关(r=0.423,0.514,0.542,0.399,P均<0.05); 血清miR-497、TNF-α对ACI诊断的曲线下面积(Area under curve,AUC)为0.848、0.806,截断值分别为1.29、1.27,相应灵敏度分别为82.3%、81.3%,特异度分别为76.5%、77.6%; 两者联合诊断ACI的AUC为0.907,其灵敏度、特异度分别为81.3%、90.8%。结论 miR-497、TNF-α在ACI患者血清中表达均上调,且与神经功能缺损程度、脑梗死体积有关,均可能在ACI进展中起一定作用,两者联合可有效提高ACI的诊断效能,有助于诊断、评估ACI患者的病情。     

CD95-mediated apoptosis and DNA fragmentation in MS

OBJECTIVE: To investigate possible associations of soluble CD95 (sCD95) serum levels and DNA defragmentation with different MS disease stages and activities. METHODS: Sera of 114 patients were analysed by an ELISA technique for sCD95. In a subgroup of 18 relapsing-remitting MS patients and controls we studied DNA fragmentation by the TUNEL-method in CSF cytospins. RESULTS: sCD95 was detectable in sera of MS patients, healthy controls and meningitis patients without significant differences. CSF specimens showed modest amounts of apoptotic cells in MS and controls. CONCLUSION: We could not demonstrate an association of MS disease course or activity with the expression of sCD95 in sera. DNA fragmentation in the CSF was not significantly enhanced compared to controls. Thus the analysed markers of programmed cell death appear not suitable to monitor MS disease courses.     

Serum uric acid levels in multiple sclerosis patients correlate with activity of disease and blood–brain barrier dysfunction

Several findings suggest lower levels of serum uric acid in multiple sclerosis (MS) patients. The aim of this study is to investigate relationships of uric acid serum levels in relapse-remitting (RR) MS patients with clinical activity of disease and blood-brain barrier (BBB) condition. Sixty-three definite RRMS patients and 40 controls divided into two groups: 20 healthy donors and 20 patients with other inflammatory neurological diseases (OINDs) were analysed. By using a quantitative enzymatic assay according to the manufacture's protocol and a commercial uric acid standard solution, serum uric acid levels were measured and the results were standardized. To investigate BBB function, magnetic resonance imaging after administration of gadolinium was used. MS patients were found to have significantly lower serum uric acid levels (193.89 +/- 49.05 micromol/l; mean value +/-SD) in comparison with healthy donors (292.7 +/- 58.65 micromol/l; P=0.000) and OIND patients (242.7 +/- 46.66 micromol/l; P=0.001). We found that MS patients with relapse had significantly lower serum uric acid levels (161.49 +/- 23.61 micromol/l) than MS patients with remission (234.39 +/- 41.96 micromol/l; P=0.000) and more over, MS patients with BBB disruption had significantly lower serum uric acid levels (163.95 +/- 26.07 micromol/l) than those with normal BBB (252.48 +/- 25.94 micromol/l; P=0.000). Further, we also found that serum uric acid level independently correlated with disease activity, BBB disruption, and gender. These results indicate that lower uric acid levels in MS patients are associated with relapse and suggest that uric acid might be beneficial in the treatment of MS.     

TNF–α converting enzyme (TACE) protein expression in different clinical subtypes of multiple sclerosis

Tumor necrosis factor (TNF)–α converting enzyme (TACE, also called ADAM17) is a key sheddase that releases TNF–α from its inactive cell–bound precursor. TACE protein expression levels in peripheral blood mononuclear cells were measured by Western blot analysis in 20 healthy controls and 80 multiple sclerosis (MS) patients before and after treatment with IFNβ [20 patients with primary progressive (PP) MS, 20 patients with secondary progressive (SP) MS, and 40 patients with relapsing– remitting (RR) MS (20 patients during clinical remission and 20 patients in relapse)]. TNF–α serum levels were also measured by enzyme–linked immunoassay in the MS patients and healthy controls. TACE protein expression levels were lower in healthy controls and PPMS patients compared with SPMS patients and RRMS patient during clinical remission. No differences in TACE protein levels were observed between RRMS patients in relapse and during remission. TACE protein levels were increased in PPMS patients treated with IFNβ. Serum TNF–α levels were higher in RRMS patients in relapse compared with RRMS patients during remission, and positive and negative correlations were found between TACE protein expression and serum TNF–α levels in RRMS patients during relapse and during remission respectively. These findings point to different regulatory mechanisms of the TACE–TNF–α pathway in the clinical MS subtypes and expand the role of TACE in MS pathogenesis.     

Lack of association of the CD14/C -159T polymorphism with susceptibility and progression parameters in Turkish multiple sclerosis patients

Soluble (s) CD14, being a receptor for lipopolysaccharides (LPSs) may inhibit LPS-triggered apoptosis and T lymphocyte proliferation. C to T exchange at position -159 in the promoter region of the CD14 gene might lead to higher sCD14 levels. Limited number of groups have studied whether these polymorphisms might influence the development of organ specific autoimmunity and whether higher CD14 levels are associated with increased levels of cytokines trigerring inflammatory processes. However their data contradict each other. In this study serum levels of sCD14 based on ELISA were measured in 77 treatment-naive patients and in 67 healthy controls. As the C-159T proximal promoter region regulates sCD14 levels, we investigated whether C-159T polymorphism is related to progression index in 250 MS patients vs. 183 healthy controls. CD14 polymorphism frequency between the healthy controls and the MS patients were not significantly different. While TT genotype of MS patients demonstrated significantly lower sCD14 levels compared to CC genotype; this difference was not reflected on the disease progression index. Our study that extends the prior data of previous studies reflects that sCD14 do not appear to be a solely prominent element of innate immunity in MS.     

Expression of B-cell-activating factor of the TNF family (BAFF) and its receptors in multiple sclerosis

The role of B cells and antibodies in the pathogenesis of multiple sclerosis (MS) is controversial. We investigated the expression of B-cell-activating factor of the tumor necrosis factor family (BAFF), a protein indispensable for B-cell survival, and of its three receptors in MS patients and controls. BAFF mRNA levels in monocytes, and BAFF-receptor mRNA in B and T cells, were higher in patients than in healthy controls; yet, BAFF protein levels in cerebrospinal fluid and plasma were similar in patients and headache controls. In addition, each MS disease course was associated with a unique expression pattern for all four molecules.     

Circadian rhythmicity of inflammatory serum parameters: a neglected issue in the search of biomarkers in multiple sclerosis

Inflammatory serum parameters are intensely investigated in the search of biomarkers for disease activity and treatment response in multiple sclerosis (MS). A reason for contradictory results might be the timing of blood collection for analyzing serum concentrations of inflammatory parameters which are subject to diurnal changes. We included 34 untreated patients with relapsing-remitting MS and 34 age- and sex-matched healthy controls. 12 MS patients showed acute disease activity in corresponding MRI scans. Blood samples were obtained at 7.00, 11.00 am, 2.30, 6.00 and 9.30 pm within 1 day. We determined serum levels of cortisol and inflammatory markers including soluble tumor necrosis factor-beta (sTNF-β), soluble TNF-Receptor-1 (sTNF-R1) and -2 (sTNF-2), soluble vascular adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-1 (sICAM-1) by ELISA. We observed significantly higher serum levels of sTNF-R1 (p < 0.001) and sTNF-R2 (p < 0.001) in the morning and a significant decline of sICAM-1 (p < 0.005) and sVCAM-1 (p < 0.001) in the afternoon in both, MS patients and healthy controls. Comparison of diurnal serum levels between MS patients with active versus with non-active disease revealed significantly higher serum levels of sVCAM-1 (p < 0.05) around noon and in the early afternoon in MS patients with active disease. A significant decline of sICAM-1 (p < 0.05) in the afternoon was seen in MS patients with active and non-active disease. Our data indicate that increased awareness of potential diurnal serum concentration changes of biomarkers can eliminate one major cause of biased data as they occur in most of the investigated immunological parameters.     

重症肌无力患者血清中miR150-5p与IL-10、IL-17水平的相关性研究

目的 检测miR150-5p在重症肌无力患者血清中的表达水平并分析与炎症因子水平的相关性。方法 收集符合本研究要求的重症肌无力患者62例和体检的健康志愿者30例,实时荧光定量聚合酶链反应(qRT-PCR)检测患者血清中miR150-5p的相对表达水平; 用酶联免疫吸附试验(ELISA)检测患者血清中炎症因子的水平; 采用pearson相关性分析法分析miR-150-5p与炎症因子水平的相关性。结果 重症肌无力患者血清中miR-150-5p的相对表达水平明显高于对照组(P<0.001); 重症肌无力患者血清中IL-2和IL-17的水平低于对照组(P<0.01),IL-10的水平高于对照组(P<0.01),而IL-19、IL-20和IL-35的水平与对照组比较无明显差异(P>0.05); 重症肌无力患者血清中miR-150-5p与IL-10的水平呈正相关(r=0.891,P<0.001),与IL-17的水平呈负相关(r=-0.836,P<0.001); 经过药物治疗后重症肌无力患者血清中miR-150-5p和IL-10的水平有所降低(P<0.001),而IL-2和IL-17的水平有所升高(P<0.001)。结论 重症肌无力患者中miR-150-5p的相对表达水平增加,且与IL-10的水平呈正相关,IL-17的水平呈负相关